U.S. patent application number 17/178151 was filed with the patent office on 2021-06-17 for substituted nucleosides, nucleotides and analogs thereof.
This patent application is currently assigned to Janssen BioPharma, Inc.. The applicant listed for this patent is Janssen BioPharma, Inc.. Invention is credited to Leonid BEIGELMAN, Jerome DEVAL, Marija PRHAVC, David Bernard SMITH, Guangyi WANG.
Application Number | 20210179656 17/178151 |
Document ID | / |
Family ID | 1000005417581 |
Filed Date | 2021-06-17 |
United States Patent
Application |
20210179656 |
Kind Code |
A1 |
BEIGELMAN; Leonid ; et
al. |
June 17, 2021 |
SUBSTITUTED NUCLEOSIDES, NUCLEOTIDES AND ANALOGS THEREOF
Abstract
Disclosed herein are nucleosides, nucleotides and analogs
thereof, pharmaceutical compositions that include one or more of
nucleosides, nucleotides and analogs thereof, and methods of
synthesizing the same. Also disclosed herein are methods of
ameliorating and/or treating a disease and/or a condition,
including an infection from a paramyxovirus and/or an
orthomyxovirus, with a nucleoside, a nucleotide and an analog
thereof.
Inventors: |
BEIGELMAN; Leonid; (San
Mateo, CA) ; WANG; Guangyi; (Carlsbad, CA) ;
SMITH; David Bernard; (San Mateo, CA) ; DEVAL;
Jerome; (Pacifica, CA) ; PRHAVC; Marija;
(Encinitas, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Janssen BioPharma, Inc. |
South San Francisco |
CA |
US |
|
|
Assignee: |
Janssen BioPharma, Inc.
South San Francisco
CA
|
Family ID: |
1000005417581 |
Appl. No.: |
17/178151 |
Filed: |
February 17, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16528414 |
Jul 31, 2019 |
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17178151 |
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14790645 |
Jul 2, 2015 |
10464965 |
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16528414 |
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13721988 |
Dec 20, 2012 |
9073960 |
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14790645 |
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61613836 |
Mar 21, 2012 |
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61579560 |
Dec 22, 2011 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 45/06 20130101;
C12Y 207/07048 20130101; C07H 19/16 20130101; C07D 487/04 20130101;
C07D 405/14 20130101; A61P 31/16 20180101; C07D 473/34 20130101;
C07D 473/18 20130101; C07H 19/00 20130101; C07H 19/20 20130101;
C07H 19/06 20130101; C07D 493/04 20130101; C12N 9/127 20130101;
C07H 19/12 20130101; C07D 405/04 20130101; C07H 19/14 20130101;
C07H 19/207 20130101; C07H 19/10 20130101 |
International
Class: |
C07H 19/06 20060101
C07H019/06; C07H 19/207 20060101 C07H019/207; C07H 19/16 20060101
C07H019/16; A61P 31/16 20060101 A61P031/16; A61K 45/06 20060101
A61K045/06; C07D 405/04 20060101 C07D405/04; C07D 405/14 20060101
C07D405/14; C07D 473/18 20060101 C07D473/18; C07D 473/34 20060101
C07D473/34; C07D 487/04 20060101 C07D487/04; C07D 493/04 20060101
C07D493/04; C07H 19/00 20060101 C07H019/00; C07H 19/14 20060101
C07H019/14; C12N 9/12 20060101 C12N009/12; C07H 19/10 20060101
C07H019/10; C07H 19/12 20060101 C07H019/12; C07H 19/20 20060101
C07H019/20 |
Claims
1. A compound selected from Formula (I), Formula (II), and Formula
(III), or a pharmaceutically acceptable salt of the foregoing:
##STR00313## wherein: B.sup.1A B.sup.1B and B.sup.1C are
independently an optionally substituted heterocyclic base or an
optionally substituted heterocyclic base with a protected amino
group; R.sup.1A is selected from the group consisting of hydrogen,
an optionally substituted acyl, an optionally substituted O-linked
amino acid, ##STR00314## when the dashed line () of Formula (I) is
a single bond, R.sup.2A is CH.sub.2, and R.sup.3A is O (oxygen);
when the dashed line () of Formula (I) is absent, R.sup.2A is
selected from the group consisting of an optionally substituted
C.sub.1-6 alkyl, an optionally substituted C.sub.2-6 alkenyl, an
optionally substituted C.sub.2-6 alkynyl, an optionally substituted
C.sub.3-6 cycloalkyl, an optionally substituted --O--C.sub.1-6
alkyl, an optionally substituted --O--C.sub.3-6 alkynyl, an
optionally substituted --O--C.sub.3-6 alkynyl and cyano, and
R.sup.3A is selected from the group consisting of OH,
--OC(.dbd.O)R''.sup.A and an optionally substituted O-linked amino
acid; R.sup.1B is selected from the group consisting of O.sup.-,
OH, ##STR00315## an optionally substituted N-linked amino acid and
an optionally substituted N-linked amino acid ester derivative;
R.sup.1C and R.sup.2C are independently selected from the group
consisting of O.sup.-, OH, an optionally substituted C.sub.1-6
alkoxy, ##STR00316## an optionally substituted N-linked amino acid
and an optionally substituted N-linked amino acid ester derivative;
or R.sup.1C is ##STR00317## and R.sup.2C is O.sup.- or OH; R.sup.2B
and R.sup.3C are independently selected from the group consisting
of an optionally substituted C.sub.1-6 alkyl, an optionally
substituted C.sub.2-6 alkenyl, an optionally substituted C.sub.2-6
alkynyl, an optionally substituted C.sub.3-6 cycloalkyl, an
optionally substituted --O--C.sub.1-6 alkyl, an optionally
substituted --O--C.sub.3-6 alkenyl, an optionally substituted
--O--C.sub.3-6 alkynyl and cyano; R.sup.4C is selected from the
group consisting of OH, --OC(.dbd.O)R''.sup.C and an optionally
substituted O-linked amino acid; R.sup.4A, R.sup.3B and R.sup.5C
are independently a halogen; R.sup.5A, R.sup.4B and R.sup.6C are
independently hydrogen or halogen; R.sup.6A, R.sup.7A and R.sup.8A
are independently selected from the group consisting of absent,
hydrogen, an optionally substituted C.sub.1-24 alkyl, an optionally
substituted C.sub.2-24 alkenyl, an optionally substituted
C.sub.2-24 alkynyl, an optionally substituted C.sub.3-6 cycloalkyl,
an optionally substituted C.sub.3-6 cycloalkenyl, an optionally
substituted aryl, an optionally substituted heteroaryl, an
optionally substituted aryl(C.sub.1-6 alkyl), an optionally
substituted *--(CR.sup.15AR.sup.16A).sub.p--O--C.sub.1-24 alkyl, an
optionally substituted *--(CR.sup.17AR.sup.1A).sub.q--O--C.sub.1-24
alkenyl, ##STR00318## R.sup.6A is ##STR00319## and R.sup.7A is
absent or hydrogen; or R.sup.6A and R.sup.7A are taken together to
form a moiety selected from the group consisting of an optionally
substituted ##STR00320## and an optionally substituted ##STR00321##
wherein the oxygens connected to R.sup.6A and R.sup.7A, the
phosphorus and the moiety form a six-membered to ten-membered ring
system; R.sup.9A is independently selected from the group
consisting of an optionally substituted C.sub.1-24 alkyl, an
optionally substituted C.sub.2-24 alkenyl, an optionally
substituted C.sub.2-24 alkynyl, an optionally substituted C.sub.3-6
cycloalkyl, an optionally substituted C.sub.3-6 cycloalkenyl,
NR.sup.30AR.sup.31A, an optionally substituted N-linked amino acid
and an optionally substituted N-linked amino acid ester derivative;
R.sup.10A and R.sup.11A are independently an optionally substituted
N-linked amino acid or an optionally substituted N-linked amino
acid ester derivative; R.sup.12A, R.sup.13A and R.sup.14A are
independently absent or hydrogen; each R.sup.15A, each R.sup.16A,
each R.sup.17A and each R.sup.18A are independently hydrogen, an
optionally substituted C.sub.1-24 alkyl or alkoxy; R.sup.19A,
R.sup.20A, R.sup.22A, R.sup.23A, R.sup.5B, R.sup.6B, R.sup.8B,
R.sup.9B, R.sup.9C, R.sup.10C, R.sup.12C and R.sup.13C are
independently selected from the group consisting of hydrogen, an
optionally substituted C.sub.1-24 alkyl and an optionally
substituted aryl; R.sup.21A, R.sup.24A, R.sup.7B, R.sup.10B,
R.sup.11C and R.sup.14C are independently selected from the group
consisting of hydrogen, an optionally substituted C.sub.1-24 alkyl,
an optionally substituted aryl, an optionally substituted
--O--C.sub.1-24 alkyl and an optionally substituted --O-aryl;
R.sup.25A, R.sup.29A, R.sup.11B and R.sup.15C are independently
selected from the group consisting of hydrogen, an optionally
substituted C.sub.1-24 alkyl and an optionally substituted aryl;
R.sup.16C, R.sup.17C and R.sup.18C are independently absent or
hydrogen; R.sup.26A and R.sup.27A are independently --C.ident.N or
an optionally substituted substituent selected from the group
consisting of C.sub.2-8 organylcarbonyl, C.sub.2-8 alkoxycarbonyl
and C.sub.2-8 organylaminocarbonyl; R.sup.28A is selected from the
group consisting of hydrogen, an optionally substituted
C.sub.1-24-alkyl, an optionally substituted C.sub.2-24 alkenyl, an
optionally substituted C.sub.2-24 alkynyl, an optionally
substituted C.sub.3-6 cycloalkyl and an optionally substituted
C.sub.3-6 cycloalkenyl; R.sup.30A and R.sup.31A are independently
selected from the group consisting of hydrogen, an optionally
substituted C.sub.1-24-alkyl, an optionally substituted C.sub.2-24
alkenyl, an optionally substituted C.sub.2-24 alkynyl, an
optionally substituted C.sub.3-6 cycloalkyl and an optionally
substituted C.sub.3-6 cycloalkenyl; for Formula (III), is a single
bond or a double bond; when is a single bond, each R.sup.7C and
each R.sup.8C is independently hydrogen or halogen; and when is a
double bond, each R.sup.7C is absent and each R.sup.8C is
independently hydrogen or halogen; R''.sup.A and R.sup.''C are
independently an optionally substituted C.sub.1-24-alkyl; m and n
are independently 0 or 1; p and q are independently selected from
the group consisting of 1, 2 and 3; r is 1 or 2; Z.sup.1A,
Z.sup.2A, Z.sup.3A, Z.sup.4A, Z.sup.1B, Z.sup.2B and Z.sup.1C are
independently O or S; and provided that when the dashed line () of
Formula (I) is absent; R.sup.1A is ##STR00322## wherein R.sup.8A is
an unsubstituted C.sub.1-4 alkyl or phenyl optionally
para-substituted with a halogen or methyl and R.sup.9A is methyl
ester, ethyl ester, isopropyl ester, n-butyl ester, benzyl ester or
phenyl ester of an amino acid selected from the group consisting of
glycine, alanine, valine, leucine, phenylalanine, tryptophan,
methionine and proline; R.sup.3A is OH; R.sup.4A is fluoro;
R.sup.5A is fluoro or hydrogen; and B.sup.1A is an unsubstituted
uracil; then R.sup.2A cannot be --OCH.sub.3; provided that when the
dashed line () of Formula (I) is absent; R.sup.1A is H; R.sup.3A is
OH; R.sup.4A is fluoro; R.sup.5A is fluoro; and B.sup.1A is an
unsubstituted cytosine; then R.sup.2A cannot be allenyl; provided
that when the dashed line () of Formula (I) is absent; R.sup.1A is
H; R.sup.3A is OH; R.sup.4A is fluoro; R.sup.5A is hydrogen; and
B.sup.1A is an unsubstituted thymine; then R.sup.2A cannot be
C.sub.1 alkyl substituted with an N-amido; and provided that when
the dashed line () of Formula (I) is absent; R.sup.1A is H;
R.sup.3A is OH; R.sup.4A is fluoro; R.sup.5A is fluoro; and
B.sup.1A is an unsubstituted cytosine; then R.sup.2A cannot be
ethynyl.
2. The compound of claim 1, wherein R.sup.1A is ##STR00323##
3. The compound of claim 1, wherein R.sup.1A is an optionally
substituted acyl.
4. The compound of claim 1, wherein R.sup.1A is H.
5. The compound of claim 1, wherein R.sup.1A is an optionally
substituted O-linked amino acid.
6. The compound of claim 1, wherein R.sup.1A is ##STR00324##
7. The compound of claim 6, wherein both R.sup.6A and R.sup.7A are
independently selected from the group consisting of an optionally
substituted C.sub.1-24 alkyl, an optionally substituted C.sub.2-24
alkenyl, an optionally substituted C.sub.2-24 alkynyl, an
optionally substituted C.sub.3-6 cycloalkyl, an optionally
substituted C.sub.3-6 cycloalkenyl, an optionally substituted aryl,
an optionally substituted heteroaryl and an optionally substituted
aryl(C.sub.1-6 alkyl).
8. The compound of claim 6, wherein both R.sup.6A and R.sup.7A are
independently selected from the group consisting of
##STR00325##
9. The compound of claim 6, wherein R.sup.6A and R.sup.7A are both
##STR00326## *--(CR.sup.15AR.sup.16A).sub.p--O--C.sub.1-24 alkyl or
*--(CR.sup.17AR.sup.18A).sub.q--O--C.sub.2-24 alkenyl; or wherein
R.sup.6A and R.sup.7A can be taken together to form a moiety
selected from the group consisting of an optionally substituted
##STR00327## and an optionally substituted ##STR00328## wherein the
oxygens connected to R.sup.6A and R.sup.7A, the phosphorus and the
moiety form a six-membered to ten-membered ring system.
10. The compound of claim 6, wherein R.sup.8A is selected from the
group consisting of absent, hydrogen, an optionally substituted
C.sub.1-24 alkyl, an optionally substituted C.sub.2-24 alkenyl, an
optionally substituted C.sub.2-24 alkynyl, an optionally
substituted C.sub.3-6 cycloalkyl and an optionally substituted
C.sub.3-6 cycloalkenyl; and R.sup.9A is independently selected from
the group consisting of an optionally substituted C.sub.1-24 alkyl,
an optionally substituted C.sub.2-24 alkenyl, an optionally
substituted C.sub.2-24 alkynyl, an optionally substituted C.sub.3-6
cycloalkyl, an optionally substituted C.sub.3-6 cycloalkenyl and
NR.sup.30AR.sup.31A.
11. The compound of claim 6, wherein R.sup.8A is an optionally
substituted aryl; and R.sup.9A is an optionally substituted
N-linked amino acid or an optionally substituted N-linked amino
acid ester derivative.
12. The compound of claim 6, wherein R.sup.10A and R.sup.11A are
both an optionally substituted N-linked amino acid or an optionally
substituted N-linked amino acid ester derivative.
13. The compound of claim 1, wherein B.sup.1A is selected from the
group consisting of: ##STR00329## wherein: R.sup.A2 is selected
from the group consisting of hydrogen, halogen and NHR.sup.J2,
wherein R.sup.J2 is selected from the group consisting of hydrogen,
--C(.dbd.O)R.sup.K2 and --C(.dbd.O)OR.sup.L2; R.sup.B2 is halogen
or NHR.sup.W2, wherein R.sup.W2 is selected from the group
consisting of hydrogen, an optionally substituted C.sub.1-6 alkyl,
an optionally substituted C.sub.2-6 alkenyl, an optionally
substituted C.sub.3-8 cycloalkyl, --C(.dbd.O)R.sup.M2 and
--C(.dbd.O)OR.sup.N2; R.sup.C2 is hydrogen or NHR.sup.O2, wherein
R.sup.O2 is selected from the group consisting of hydrogen,
--C(.dbd.O)R.sup.P2 and --C(.dbd.O)OR.sup.Q2; R.sup.D2 is selected
from the group consisting of hydrogen, halogen, an optionally
substituted C.sub.1-6 alkyl, an optionally substituted C.sub.2-6
alkenyl and an optionally substituted C.sub.2-6 alkynyl; R.sup.E2
is selected from the group consisting of hydrogen, hydroxy, an
optionally substituted C.sub.1-6 alkyl, an optionally substituted
C.sub.3-8 cycloalkyl, --C(.dbd.O)R.sup.R2 and --C(.dbd.O)OR.sup.S2;
R.sup.F2 is selected from the group consisting of hydrogen,
halogen, an optionally substituted C.sub.1-6alkyl, an optionally
substituted C.sub.2-6 alkenyl and an optionally substituted
C.sub.2-6 alkynyl; Y.sup.2 and Y.sup.3 are independently N or
CR.sup.I2, wherein R.sup.I2 is selected from the group consisting
of hydrogen, halogen, an optionally substituted C.sub.1-6-alkyl, an
optionally substituted C.sub.2-6-alkenyl and an optionally
substituted C.sub.2-6-alkynyl; R.sup.G2 is an optionally
substituted C.sub.1-6 alkyl; R.sup.H2 is hydrogen or NHR.sup.T2,
wherein R.sup.T2 is independently selected from the group
consisting of hydrogen, --C(.dbd.O)R.sup.U2 and
--C(.dbd.O)OR.sup.V2; and R.sup.K2, R.sup.L2, R.sup.M2, R.sup.N2,
R.sup.P2, R.sup.Q2 R.sup.R2, R.sup.S2, R.sup.U2 and R.sup.V2 are
independently selected from the group consisting of C.sub.1-6
alkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.3-6 cycloalkyl,
C.sub.3-6 cycloalkenyl, C.sub.6-10 aryl, heteroaryl,
heteroalicyclyl, aryl(C.sub.1-6 alkyl), heteroaryl(C.sub.1-6 alkyl)
and heteroalicyclyl(C.sub.1-6 alkyl).
14. The compound of claim 1, wherein the dashed line (------) is
absent, R.sup.2A is selected from the group consisting of an
optionally substituted C.sub.1-6 alkyl, an optionally substituted
C.sub.2-6 alkenyl, an optionally substituted C.sub.2-6 alkynyl, an
optionally substituted --O--C.sub.1-6 alkyl, an optionally
substituted --O--C.sub.3-6 alkenyl, an optionally substituted
--O--C.sub.3-6 alkynyl and cyano, and R.sup.3A is selected from the
group consisting of OH, --OC(.dbd.O)R.sup.''A and an optionally
substituted O-linked amino acid.
15. The compound of claim 1, wherein the compound of Formula (I) is
selected from the group consisting of: ##STR00330## ##STR00331##
##STR00332## or a pharmaceutically acceptable salt thereof.
16. The compound of claim 1, wherein the compound of Formula (I) is
selected from the group consisting of: ##STR00333## ##STR00334##
##STR00335## ##STR00336## ##STR00337## ##STR00338## ##STR00339##
##STR00340## ##STR00341## ##STR00342## or a pharmaceutically
acceptable salt of the foregoing.
17. The compound of claim 1, wherein the compound of Formula (I) is
selected from the group consisting of: ##STR00343## ##STR00344##
##STR00345## ##STR00346## ##STR00347## ##STR00348## or a
pharmaceutically acceptable salt of the foregoing.
18. The compound of claim 1, wherein the compound of Formula (I) is
selected from the group consisting of: ##STR00349## ##STR00350##
##STR00351## ##STR00352## ##STR00353## ##STR00354## ##STR00355## or
a pharmaceutically acceptable salt of the foregoing.
19. A pharmaceutical composition comprising an effective amount of
a compound of claim 1, or a pharmaceutically acceptable salt
thereof, and a pharmaceutically acceptable carrier, diluent,
excipient, or combination thereof.
20. A method for ameliorating or treating a viral infection
comprising contacting a cell infected with the virus in a subject
identified as suffering from the viral infection with an effective
amount of a compound of claim 1, or a pharmaceutically acceptable
salt thereof; and wherein the viral infection is selected from a
paramyxovirus viral infection and an orthomyxovirus viral
infection.
21. A method for ameliorating or treating a viral infection in
combination with one or more agents comprising administering to or
contacting a cell in a subject identified as suffering from the
viral infection with an effective amount of a compound of claim 1,
or a pharmaceutically acceptable salt thereof; and wherein the
viral infection is selected from a paramyxovirus viral infection
and an orthomyxovirus viral infection.
Description
[0001] This application is a continuation of U.S. application Ser.
No. 13/721,988, filed Dec. 20, 2012, which claims the benefit of
U.S. Provisional Application Nos. 61/579,560, filed Dec. 22, 2011;
and 61/613,836, filed Mar. 21, 2012; which are incorporated herein
by reference in their entireties; including any drawings.
BACKGROUND
Field
[0002] The present application relates to the fields of chemistry,
biochemistry and medicine. More particularly, disclosed herein are
nucleoside, nucleotides and analogs thereof, pharmaceutical
compositions that include one or more nucleosides, nucleotides and
analogs thereof, and methods of synthesizing the same. Also
disclosed herein are methods of ameliorating and/or treating a
paramyxovirus and/or an orthomyxovirus viral infection with one or
more nucleosides, nucleotides and analogs thereof.
Description
[0003] Respiratory viral infections, including upper and lower
respiratory tract viral infections, infects and is the leading
cause of death of millions of people each year. Upper respiratory
tract viral infections involve the nose, sinuses, pharynx and/or
larynx. Lower respiratory tract viral infections involve the
respiratory system below the vocal cords, including the trachea,
primary bronchi and lungs.
[0004] Nucleoside analogs are a class of compounds that have been
shown to exert antiviral activity both in vitro and in vivo, and
thus, have been the subject of widespread research for the
treatment of viral infections. Nucleoside analogs are usually
therapeutically inactive compounds that are converted by host or
viral enzymes to their respective active anti-metabolites, which,
in turn, may inhibit polymerases involved in viral or cell
proliferation. The activation occurs by a variety of mechanisms,
such as the addition of one or more phosphate groups and, or in
combination with, other metabolic processes.
SUMMARY
[0005] Some embodiments disclosed herein relate to a compound of
Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing.
[0006] Some embodiments disclosed herein relate to methods of
ameliorating and/or treating a paramyxovirus viral infection that
can include administering to a subject suffering from the
paramyxovirus viral infection an effective amount of one or more
compounds of Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing, or a
pharmaceutical composition that includes one or more compounds of
Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing. Other
embodiments described herein relate to using one or more compounds
of Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing, in the
manufacture of a medicament for ameliorating and/or treating a
paramyxovirus viral infection. Still other embodiments described
herein relate to compounds of Formula (I), Formula (II) and/or
Formula (III), or a pharmaceutically acceptable salt of the
foregoing, that can be used for ameliorating and/or treating a
paramyxovirus viral infection. Yet still other embodiments
disclosed herein relate to methods of ameliorating and/or treating
a paramyxovirus viral infection that can include contacting a cell
infected with the paramyxovirus with an effective amount of one or
more compounds of Formula (I), Formula (II) and/or Formula (III),
or a pharmaceutically acceptable salt of the foregoing, or a
pharmaceutical composition that includes one or more compounds of
Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing. Some embodiments
disclosed herein relate to methods of inhibiting the replication of
a paramyxovirus that can include contacting a cell infection with
the paramyxovirus with an effective amount of one or more compounds
of Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing, or a
pharmaceutical composition that includes one or more compounds of
Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing. For example, the
paramyxovirus viral infection can be caused by a henipavirus, a
morbillivirus, a respirovirus, a rubulavirus, a pneumovirus
(including a respiratory syncytial viral infection), a
metapneumovirus, hendravirus, nipahvirus, measles, sendai virus,
mumps, a human parainfluenza virus (HPIV-1, HPIV-2, HPIV-3 and
HPIV-4) and/or a metapneumovirus.
[0007] Some embodiments disclosed herein relate to methods of
ameliorating and/or treating an orthomyxovirus viral infection that
can include administering to a subject suffering from the
orthomyxovirus viral infection an effective amount of one or more
compounds of Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing, or a
pharmaceutical composition that includes one or more compounds of
Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing. Other
embodiments described herein relate to using one or more compounds
of Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing, in the
manufacture of a medicament for ameliorating and/or treating an
orthomyxovirus viral infection. Still other embodiments described
herein relate to compounds of Formula (I), Formula (II) and/or
Formula (III), or a pharmaceutically acceptable salt of the
foregoing, that can be used for ameliorating and/or treating an
orthomyxovirus viral infection. Yet still other embodiments
disclosed herein relate to methods of ameliorating and/or treating
an orthomyxovirus viral infection that can include contacting a
cell infected with the orthomyxovirus with an effective amount of
one or more compounds of Formula (I), Formula (II) and/or Formula
(III), or a pharmaceutically acceptable salt of the foregoing, or a
pharmaceutical composition that includes one or more compounds of
Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing. Some embodiments
disclosed herein relate to methods of inhibiting the replication of
an orthomyxovirus that can include contacting a cell infection with
the orthomyxovirus with an effective amount of one or more
compounds of Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing, or a
pharmaceutical composition that includes one or more compounds of
Formula (I), Formula (II) and/or Formula (III), or a
pharmaceutically acceptable salt of the foregoing. For example, the
orthomyxovirus viral infection can be an influenza viral infection
(such as influenza A, B and/or C).
[0008] Some embodiments disclosed herein relate to methods of
ameliorating and/or treating a paramyxovirus viral infection and/or
an orthomyxovirus viral infection that can include administering to
a subject suffering from the viral infection an effective amount of
a compound described herein or a pharmaceutically acceptable salt
thereof (for example, one or more compounds of Formulae (I), (II)
and/or (III), or a pharmaceutically acceptable salt of the
foregoing), or a pharmaceutical composition that includes one or
more compounds described herein, in combination with one or more
agents described herein. Some embodiments disclosed herein relate
to methods of ameliorating and/or treating a paramyxovirus viral
infection and/or an orthomyxovirus viral infection that can include
contacting a cell infected with the virus with an effective amount
of a compound described herein or a pharmaceutically acceptable
salt thereof (for example, one or more compounds of Formulae (I),
(II) and/or (III), or a pharmaceutically acceptable salt of the
foregoing), or a pharmaceutical composition that includes one or
more compounds described herein, in combination with one or more
agents described herein.
BRIEF DESCRIPTION OF THE DRAWINGS
[0009] FIG. 1 shows example RSV agents.
DETAILED DESCRIPTION
[0010] Paramyxoviridae family is a family of single stranded RNA
viruses. Several genera of the paramyxoviridae family include
henipavirus, morbillivirus, respirovirus, rubulavirus, pneumovirus
and metapneumovirus. These viruses can be transmitted person to
person via direct or close contact with contaminated respiratory
droplets or fomites. Species of henipavirus include hendravirus and
nipahvirus. A species of morbillivirus is measles. Species of
respirovirus include sendai virus and human parainfluenza viruses 1
and 3; and species of rubulavirus include mumps virus and human
parainfluenza viruses 2 and 4. A species of metapneumovirus is
human metapneumovirus.
[0011] Human Respiratory Syncytial Virus (RSV), a species of
pneumovirus, can cause respiratory infections, and can be
associated with bronchiolitis and pneumonia. Symptoms of an RSV
infection include coughing, sneezing, runny nose, fever, decrease
in appetite, and wheezing. RSV is the most common cause of
bronchiolitis and pneumonia in children under one year of age in
the world, and can be the cause of tracheobronchitis in older
children and adults. In the United States, between 75,000 and
125,000 infants are hospitalized each year with RSV. Among adults
older than 65 years of age, an estimated 14,000 deaths and 177,000
hospitalizations have been attributed to RSV.
[0012] Treatment options for people infected with RSV are currently
limited. Antibiotics, usually prescribed to treat bacterial
infections, and over-the-counter medication are not effective in
treating RSV and may help only to relieve some of the symptoms. In
severe cases, a nebulized bronchodilator, such as albuterol, may be
prescribed to relieve some of the symptoms, such as wheezing.
RespiGram.RTM. (RSV-IGIV, MedImmune, approved for high risk
children younger than 24 months of age), Synagis.RTM. (palivizumab,
MedImmune, approved for high risk children younger than 24 months
of age), and Virzole.RTM. (ribavirin by aerosol, ICN
pharmaceuticals) have been approved for treatment of RSV.
[0013] Symptoms of the measles include fever, cough, runny nose,
red eyes and a generalized rash. Some individuals with measles can
develop pneumonia, ear infections and bronchitis. Mumps leads to
swelling of the salivary glands. Symptoms of mumps include fever,
loss of appetite and fatigue. Individuals are often immunized
against measles and mumps via a three-part MMR vaccine (measles,
mumps, and rubella). Human parainfluenza virus includes four
serotypes types, and can cause upper and lower respiratory tract
infections. Human parainfluenza virus 1 (HPIV-1) can be associated
with croup; human parainfluenza virus 3 (HPIV-3) can be associated
with bronchiolitis and pneumonia. According to the Centers of
Disease Control and Prevention (CDC), there are no vaccines against
human parainfluenza virus.
[0014] Influenza is a single stranded RNA virus and a member of the
Orthomyxoviridae family. There are currently three species of
influenza; influenza A, influenza B and influenza C. Influenza A
has been further classified based on the viral surface proteins
into hemagglutinin (H or HA) and neuramididase (N). There are
approximately 16 H antigens (H1 to H16) and 9 N antigens (N1 to
N9). Influenza A includes several subtype, including H1N1, H1N2,
H2N2, H3N1, H3N2, H3N8, H5N1, H5N2, H5N3, H5N8, H5N9, H7N1, H7N2,
H7N3, H7N4, H7N7, H9N2, H10N7. As with RSV, influenza viruses can
be transmitted from person to person via direct contact with
infected secretions and/or contaminated surfaces or objections.
Complications from an influenza viral infection include pneumonia,
bronchitis, dehydration, and sinus and ear infections. Medications
currently approved by the FDA against an influenza infection
include amantadine, rimantadine, Relenza.RTM. (zanamivir,
GlaxoSmithKline) and Tamiflu.RTM. (oseltamivir, Genentech).
Definitions
[0015] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as is commonly understood by one
of ordinary skill in the art. All patents, applications, published
applications and other publications referenced herein are
incorporated by reference in their entirety unless stated
otherwise. In the event that there are a plurality of definitions
for a term herein, those in this section prevail unless stated
otherwise.
[0016] As used herein, any "R" group(s) such as, without
limitation, R.sup.1A, R.sup.2A, R.sup.3A, R.sup.4A, R.sup.5A,
R.sup.6A, R.sup.7A, R.sup.8A, R.sup.9A, R.sup.10A, R.sup.11A,
R.sup.12A, R.sup.13A, R.sup.14A, R.sup.15A, R.sup.16A, R.sup.17A,
R.sup.18A, R.sup.19A, R.sup.20A, R.sup.21A, R.sup.22A, R.sup.23A,
R.sup.24A, R.sup.25A, R.sup.26A, R.sup.27A, R.sup.28A, R.sup.29A,
R.sup.30A, R.sup.31A, R.sup.32A, R.sup.33A, R.sup.34A, R.sup.35A,
R.sup.36A, R.sup.37A, R.sup.38A, R.sup.1B, R.sup.2B, R.sup.3B,
R.sup.4B, R.sup.5B, R.sup.6B, R.sup.7B, R.sup.8B, R.sup.9B,
R.sup.10B, R.sup.11B, R.sup.12B, R.sup.13B, R.sup.14B, R.sup.1C,
R.sup.2C, R.sup.3C, R.sup.4C, R.sup.5C, R.sup.6C, R.sup.7C,
R.sup.8C, R.sup.9C, R.sup.10C, R.sup.11C, R.sup.12C, R.sup.13C,
R.sup.14C, R.sup.15C, R.sup.16C, R.sup.17C, R.sup.18C, R.sup.19C,
R.sup.20C, R.sup.21C, R.sup.22C and R.sup.23C represent
substituents that can be attached to the indicated atom. An R group
may be substituted or unsubstituted. If two "R" groups are
described as being "taken together" the R groups and the atoms they
are attached to can form a cycloalkyl, cycloalkenyl, cycloalkynyl,
aryl, heteroaryl or heterocycle. For example, without limitation,
if R.sup.a and R.sup.b of an NR.sup.a R.sup.b group are indicated
to be "taken together," it means that they are covalently bonded to
one another to form a ring:
##STR00001##
In addition, if two "R" groups are described as being "taken
together" with the atom(s) to which they are attached to form a
ring as an alternative, the R groups are not limited to the
variables or substituents defined previously.
[0017] Whenever a group is described as being "optionally
substituted" that group may be unsubstituted or substituted with
one or more of the indicated substituents. Likewise, when a group
is described as being "unsubstituted or substituted" if
substituted, the substituent(s) may be selected from one or more
the indicated substituents. If no substituents are indicated, it is
meant that the indicated "optionally substituted" or "substituted"
group may be substituted with one or more group(s) individually and
independently selected from alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclyl,
aralkyl, heteroaralkyl, (heteroalicyclyl)alkyl, hydroxy, protected
hydroxyl, alkoxy, aryloxy, acyl, mercapto, alkylthio, arylthio,
cyano, halogen, thiocarbonyl, O-carbamyl, N-carbamyl,
O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido,
N-sulfonamido, C-carboxy, protected C-carboxy, O-carboxy,
isocyanato, thiocyanato, isothiocyanato, nitro, silyl, sulfenyl,
sulfinyl, sulfonyl, haloalkyl, haloalkoxy, trihalomethanesulfonyl,
trihalomethanesulfonamido, an amino, a mono-substituted amino group
and a di-substituted amino group, and protected derivatives
thereof.
[0018] As used herein, "C.sub.a to C.sub.b" in which "a" and "b"
are integers refer to the number of carbon atoms in an alkyl,
alkenyl or alkynyl group, or the number of carbon atoms in the ring
of a cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl or
heteroalicyclyl group. That is, the alkyl, alkenyl, alkynyl, ring
of the cycloalkyl, ring of the cycloalkenyl, ring of the
cycloalkynyl, ring of the aryl, ring of the heteroaryl or ring of
the heteroalicyclyl can contain from "a" to "b", inclusive, carbon
atoms. Thus, for example, a "C.sub.1 to C.sub.4 alkyl" group refers
to all alkyl groups having from 1 to 4 carbons, that is,
CH.sub.3--, CH.sub.3CH.sub.2--, CH.sub.3CH.sub.2CH.sub.2--,
(CH.sub.3).sub.2CH--, CH.sub.3CH.sub.2CH.sub.2CH.sub.2--,
CH.sub.3CH.sub.2CH(CH.sub.3)-- and (CH.sub.3).sub.3C--. If no "a"
and "b" are designated with regard to an alkyl, alkenyl, alkynyl,
cycloalkyl cycloalkenyl, cycloalkynyl, aryl, heteroaryl or
heteroalicyclyl group, the broadest range described in these
definitions is to be assumed.
[0019] As used herein, "alkyl" refers to a straight or branched
hydrocarbon chain that comprises a fully saturated (no double or
triple bonds) hydrocarbon group. The alkyl group may have 1 to 20
carbon atoms (whenever it appears herein, a numerical range such as
"1 to 20" refers to each integer in the given range; e.g., "to 20
carbon atoms" means that the alkyl group may consist of 1 carbon
atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20
carbon atoms, although the present definition also covers the
occurrence of the term "alkyl" where no numerical range is
designated). The alkyl group may also be a medium size alkyl having
1 to 10 carbon atoms. The alkyl group could also be a lower alkyl
having 1 to 6 carbon atoms. The alkyl group of the compounds may be
designated as "C.sub.1-C.sub.4 alkyl" or similar designations. By
way of example only, "C.sub.1-C.sub.4 alkyl" indicates that there
are one to four carbon atoms in the alkyl chain, i.e., the alkyl
chain is selected from methyl, ethyl, propyl, iso-propyl, n-butyl,
iso-butyl, sec-butyl, and t-butyl. Typical alkyl groups include,
but are in no way limited to, methyl, ethyl, propyl, isopropyl,
butyl, isobutyl, tertiary butyl, pentyl and hexyl. The alkyl group
may be substituted or unsubstituted.
[0020] As used herein, "alkenyl" refers to an alkyl group that
contains in the straight or branched hydrocarbon chain one or more
double bonds. An alkenyl group may be unsubstituted or
substituted.
[0021] As used herein, "alkynyl" refers to an alkyl group that
contains in the straight or branched hydrocarbon chain one or more
triple bonds. An alkynyl group may be unsubstituted or
substituted.
[0022] As used herein, "cycloalkyl" refers to a completely
saturated (no double or triple bonds) mono- or multi-cyclic
hydrocarbon ring system. When composed of two or more rings, the
rings may be joined together in a fused fashion. Cycloalkyl groups
can contain 3 to 10 atoms in the ring(s) or 3 to 8 atoms in the
ring(s). A cycloalkyl group may be unsubstituted or substituted.
Typical cycloalkyl groups include, but are in no way limited to,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and
cyclooctyl.
[0023] As used herein, "cycloalkenyl" refers to a mono- or
multi-cyclic hydrocarbon ring system that contains one or more
double bonds in at least one ring; although, if there is more than
one, the double bonds cannot form a fully delocalized pi-electron
system throughout all the rings (otherwise the group would be
"aryl," as defined herein). When composed of two or more rings, the
rings may be connected together in a fused fashion. A cycloalkenyl
group may be unsubstituted or substituted.
[0024] As used herein, "cycloalkynyl" refers to a mono- or
multi-cyclic hydrocarbon ring system that contains one or more
triple bonds in at least one ring. If there is more than one triple
bond, the triple bonds cannot form a fully delocalized pi-electron
system throughout all the rings. When composed of two or more
rings, the rings may be joined together in a fused fashion. A
cycloalkynyl group may be unsubstituted or substituted.
[0025] As used herein, "aryl" refers to a carbocyclic (all carbon)
monocyclic or multicyclic aromatic ring system (including fused
ring systems where two carbocyclic rings share a chemical bond)
that has a fully delocalized pi-electron system throughout all the
rings. The number of carbon atoms in an aryl group can vary. For
example, the aryl group can be a C.sub.6-C.sub.14 aryl group, a
C.sub.6-C.sub.10 aryl group, or a C.sub.6 aryl group. Examples of
aryl groups include, but are not limited to, benzene, naphthalene
and azulene. An aryl group may be substituted or unsubstituted.
[0026] As used herein, "heteroaryl" refers to a monocyclic or
multicyclic aromatic ring system (a ring system with fully
delocalized pi-electron system) that contain(s) one or more
heteroatoms, that is, an element other than carbon, including but
not limited to, nitrogen, oxygen and sulfur. The number of atoms in
the ring(s) of a heteroaryl group can vary. For example, the
heteroaryl group can contain 4 to 14 atoms in the ring(s), 5 to 10
atoms in the ring(s) or 5 to 6 atoms in the ring(s). Furthermore,
the term "heteroaryl" includes fused ring systems where two rings,
such as at least one aryl ring and at least one heteroaryl ring, or
at least two heteroaryl rings, share at least one chemical bond.
Examples of heteroaryl rings include, but are not limited to,
furan, furazan, thiophene, benzothiophene, phthalazine, pyrrole,
oxazole, benzoxazole, 1,2,3-oxadiazole, 1,2,4-oxadiazole, thiazole,
1,2,3-thiadiazole, 1,2,4-thiadiazole, benzothiazole, imidazole,
benzimidazole, indole, indazole, pyrazole, benzopyrazole,
isoxazole, benzoisoxazole, isothiazole, triazole, benzotriazole,
thiadiazole, tetrazole, pyridine, pyridazine, pyrimidine, pyrazine,
purine, pteridine, quinoline, isoquinoline, quinazoline,
quinoxaline, cinnoline, and triazine. A heteroaryl group may be
substituted or unsubstituted.
[0027] As used herein, "heterocyclyl" or "heteroalicyclyl" refers
to three-, four-, five-, six-, seven-, eight-, nine-, ten-, up to
18-membered monocyclic, bicyclic, and tricyclic ring system wherein
carbon atoms together with from 1 to 5 heteroatoms constitute said
ring system. A heterocycle may optionally contain one or more
unsaturated bonds situated in such a way, however, that a fully
delocalized pi-electron system does not occur throughout all the
rings. The heteroatom(s) is an element other than carbon including,
but not limited to, oxygen, sulfur, and nitrogen. A heterocycle may
further contain one or more carbonyl or thiocarbonyl
functionalities, so as to make the definition include oxo-systems
and thio-systems such as lactams, lactones, cyclic imides, cyclic
thioimides and cyclic carbamates. When composed of two or more
rings, the rings may be joined together in a fused fashion.
Additionally, any nitrogens in a heteroalicyclic may be
quaternized. Heterocyclyl or heteroalicyclic groups may be
unsubstituted or substituted. Examples of such "heterocyclyl" or
"heteroalicyclyl" groups include but are not limited to,
1,3-dioxin, 1,3-dioxane, 1,4-dioxane, 1,2-dioxolane, 1,3-dioxolane,
1,4-dioxolane, 1,3-oxathiane, 1,4-oxathiin, 1,3-oxathiolane,
1,3-dithiole, 1,3-dithiolane, 1,4-oxathiane,
tetrahydro-1,4-thiazine, 2H-1,2-oxazine, maleimide, succinimide,
barbituric acid, thiobarbituric acid, dioxopiperazine, hydantoin,
dihydrouracil, trioxane, hexahydro-1,3,5-triazine, imidazoline,
imidazolidine, isoxazoline, isoxazolidine, oxazoline, oxazolidine,
oxazolidinone, thiazoline, thiazolidine, morpholine, oxirane,
piperidine N-Oxide, piperidine, piperazine, pyrrolidine,
pyrrolidone, pyrrolidione, 4-piperidone, pyrazoline, pyrazolidine,
2-oxopyrrolidine, tetrahydropyran, 4H-pyran, tetrahydrothiopyran,
thiamorpholine, thiamorpholine sulfoxide, thiamorpholine sulfone,
and their benzo-fused analogs (e.g., benzimidazolidinone,
tetrahydroquinoline, and 3,4-methylenedioxyphenyl).
[0028] As used herein, "aralkyl" and "aryl(alkyl)" refer to an aryl
group connected, as a substituent, via a lower alkylene group. The
lower alkylene and aryl group of an aralkyl may be substituted or
unsubstituted. Examples include but are not limited to benzyl,
2-phenylalkyl, 3-phenylalkyl, and naphthylalkyl.
[0029] As used herein, "heteroaralkyl" and "heteroaryl(alkyl)"
refer to a heteroaryl group connected, as a substituent, via a
lower alkylene group. The lower alkylene and heteroaryl group of
heteroaralkyl may be substituted or unsubstituted. Examples include
but are not limited to 2-thienylalkyl, 3-thienylalkyl, furylalkyl,
thienylalkyl, pyrrolylalkyl, pyridylalkyl, isoxazolylalkyl,
imidazolylalkyl, and their benzo-fused analogs.
[0030] A "(heteroalicyclyl)alkyl" and "(heterocyclyl)alkyl" refer
to a heterocyclic or a heteroalicyclic group connected, as a
substituent, via a lower alkylene group. The lower alkylene and
heterocyclyl of a (heteroalicyclyl)alkyl may be substituted or
unsubstituted. Examples include but are not limited
tetrahydro-2H-pyran-4-yl)methyl, (piperidin-4-yl)ethyl,
(piperidin-4-yl)propyl, (tetrahydro-2H-thiopyran-4-yl)methyl, and
(1,3-thiazinan-4-yl)methyl.
[0031] "Lower alkylene groups" are straight-chained --CH.sub.2--
tethering groups, forming bonds to connect molecular fragments via
their terminal carbon atoms. Examples include but are not limited
to methylene (--CH.sub.2--), ethylene (--CH.sub.2CH.sub.2--),
propylene (--CH.sub.2CH.sub.2CH.sub.2--), and butylene
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--). A lower alkylene group can
be substituted by replacing one or more hydrogen of the lower
alkylene group with a substituent(s) listed under the definition of
"substituted."
[0032] As used herein, "alkoxy" refers to the formula --OR wherein
R is an alkyl, an alkenyl, an alkynyl, a cycloalkyl, a
cycloalkenyl, a cycloalkynyl, aryl, heteroaryl, heteroalicyclyl,
aralkyl, (heteroaryl)alkyl or (heteroalicyclyl)alkyl is defined
herein. A non-limiting list of alkoxys are methoxy, ethoxy,
n-propoxy, 1-methylethoxy (isopropoxy), n-butoxy, iso-butoxy,
sec-butoxy, tert-butoxy, phenoxy and benzoxy. An alkoxy may be
substituted or unsubstituted.
[0033] As used herein, "acyl" refers to a hydrogen, alkyl, alkenyl,
alkynyl, or aryl connected, as substituents, via a carbonyl group.
Examples include formyl, acetyl, propanoyl, benzoyl, and acryl. An
acyl may be substituted or unsubstituted.
[0034] As used herein, "hydroxyalkyl" refers to an alkyl group in
which one or more of the hydrogen atoms are replaced by a hydroxy
group. Exemplary hydroxyalkyl groups include but are not limited
to, 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl, and
2,2-dihydroxyethyl. A hydroxyalkyl may be substituted or
unsubstituted.
[0035] As used herein, "haloalkyl" refers to an alkyl group in
which one or more of the hydrogen atoms are replaced by a halogen
(e.g., mono-haloalkyl, di-haloalkyl and tri-haloalkyl). Such groups
include but are not limited to, chloromethyl, fluoromethyl,
difluoromethyl, trifluoromethyl, 1-chloro-2-fluoromethyl and
2-fluoroisobutyl. A haloalkyl may be substituted or
unsubstituted.
[0036] As used herein, "haloalkoxy" refers to an alkoxy group in
which one or more of the hydrogen atoms are replaced by a halogen
(e.g., mono-haloalkoxy, di-haloalkoxy and tri-haloalkoxy). Such
groups include but are not limited to, chloromethoxy,
fluoromethoxy, difluoromethoxy, trifluoromethoxy,
1-chloro-2-fluoromethoxy and 2-fluoroisobutoxy. A haloalkoxy may be
substituted or unsubstituted.
[0037] As used herein, "arylthio" refers to RS--, in which R is an
aryl, such as, but not limited to, phenyl. An arylthio may be
substituted or unsubstituted.
[0038] A "sulfenyl" group refers to an "--SR" group in which R can
be hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,
cycloalkynyl, aryl, heteroaryl, heteroalicyclyl, aralkyl,
(heteroaryl)alkyl or (heteroalicyclyl)alkyl. A sulfenyl may be
substituted or unsubstituted.
[0039] A "sulfinyl" group refers to an "--S(.dbd.O)--R" group in
which R can be the same as defined with respect to sulfenyl. A
sulfinyl may be substituted or unsubstituted.
[0040] A "sulfonyl" group refers to an "SO.sub.2R" group in which R
can be the same as defined with respect to sulfenyl. A sulfonyl may
be substituted or unsubstituted.
[0041] An "O-carboxy" group refers to a "RC(.dbd.O)O--" group in
which R can be hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclyl,
aralkyl, (heteroaryl)alkyl or (heteroalicyclyl)alkyl, as defined
herein. An O-carboxy may be substituted or unsubstituted.
[0042] The terms "ester" and "C-carboxy" refer to a "--C(.dbd.O)OR"
group in which R can be the same as defined with respect to
O-carboxy. An ester and C-carboxy may be substituted or
unsubstituted.
[0043] A "thiocarbonyl" group refers to a "--C(.dbd.S)R" group in
which R can be the same as defined with respect to O-carboxy. A
thiocarbonyl may be substituted or unsubstituted.
[0044] A "trihalomethanesulfonyl" group refers to an
"X.sub.3CSO.sub.2--" group wherein each X is a halogen.
[0045] A "trihalomethanesulfonamido" group refers to an
"X.sub.3CS(O).sub.2N(R.sub.A)--" group wherein each X is a halogen,
and R.sub.A hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclyl,
aralkyl, (heteroaryl)alkyl or (heteroalicyclyl)alkyl.
[0046] The term "amino" as used herein refers to a --NH.sub.2
group.
[0047] As used herein, the term "hydroxy" refers to a --OH
group.
[0048] A "cyano" group refers to a "--CN" group.
[0049] The term "azido" as used herein refers to a --N.sub.3
group.
[0050] An "isocyanato" group refers to a "--NCO" group.
[0051] A "thiocyanato" group refers to a "--CNS" group.
[0052] An "isothiocyanato" group refers to an "--NCS" group.
[0053] A "mercapto" group refers to an "--SH" group.
[0054] A "carbonyl" group refers to a C.dbd.O group.
[0055] An "S-sulfonamido" group refers to a
"--SO.sub.2N(R.sub.AR.sub.B)" group in which R.sub.A and R.sub.B
can be independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclyl,
aralkyl, (heteroaryl)alkyl or (heteroalicyclyl)alkyl. An
S-sulfonamido may be substituted or unsubstituted.
[0056] An "N-sulfonamido" group refers to a "RSO.sub.2N(R.sub.A)--"
group in which R and R.sub.A can be independently hydrogen, alkyl,
alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl,
heteroaryl, heteroalicyclyl, aralkyl, (heteroaryl)alkyl or
(heteroalicyclyl)alkyl. An N-sulfonamido may be substituted or
unsubstituted.
[0057] An "O-carbamyl" group refers to a
"--OC(.dbd.O)N(R.sub.AR.sub.B)" group in which R.sub.A and R.sub.B
can be independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclyl,
aralkyl, (heteroaryl)alkyl or (heteroalicyclyl)alkyl. An O-carbamyl
may be substituted or unsubstituted.
[0058] An "N-carbamyl" group refers to an "ROC(.dbd.O)N(R.sub.A)--"
group in which R and R.sub.A can be independently hydrogen, alkyl,
alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl,
heteroaryl, heteroalicyclyl, aralkyl, (heteroaryl)alkyl or
(heteroalicyclyl)alkyl. An N-carbamyl may be substituted or
unsubstituted.
[0059] An "O-thiocarbamyl" group refers to a
"--OC(.dbd.S)--N(R.sub.AR.sub.B)" group in which R.sub.A and
R.sub.B can be independently hydrogen, alkyl, alkenyl, alkynyl,
cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl,
heteroalicyclyl, aralkyl, (heteroaryl)alkyl or
(heteroalicyclyl)alkyl. An O-thiocarbamyl may be substituted or
unsubstituted.
[0060] An "N-thiocarbamyl" group refers to an
"ROC(.dbd.S)N(R.sub.A)--" group in which R and R.sub.A can be
independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl,
cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclyl,
aralkyl, (heteroaryl)alkyl or (heteroalicyclyl)alkyl. An
N-thiocarbamyl may be substituted or unsubstituted.
[0061] A "C-amido" group refers to a "--C(.dbd.O)N(R.sub.AR.sub.B)"
group in which R.sub.A and R.sub.B can be independently hydrogen,
alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl,
aryl, heteroaryl, heteroalicyclyl, aralkyl, (heteroaryl)alkyl or
(heteroalicyclyl)alkyl. A C-amido may be substituted or
unsubstituted.
[0062] An "N-amido" group refers to a "RC(.dbd.O)N(R.sub.A)--"
group in which R and R.sub.A can be independently hydrogen, alkyl,
alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl,
heteroaryl, heteroalicyclyl, aralkyl, (heteroaryl)alkyl or
(heteroalicyclyl)alkyl. An N-amido may be substituted or
unsubstituted.
[0063] The term "halogen atom" or "halogen" as used herein, means
any one of the radio-stable atoms of column 7 of the Periodic Table
of the Elements, such as, fluorine, chlorine, bromine and
iodine.
[0064] Where the numbers of substituents is not specified (e.g.
haloalkyl), there may be one or more substituents present. For
example "haloalkyl" may include one or more of the same or
different halogens. As another example, "C.sub.1-C.sub.3
alkoxyphenyl" may include one or more of the same or different
alkoxy groups containing one, two or three atoms.
[0065] As used herein, the abbreviations for any protective groups,
amino acids and other compounds, are, unless indicated otherwise,
in accord with their common usage, recognized abbreviations, or the
IUPAC-IUB Commission on Biochemical Nomenclature (See, Biochem.
11:942-944 (1972)).
[0066] The term "nucleoside" is used herein in its ordinary sense
as understood by those skilled in the art, and refers to a compound
composed of an optionally substituted pentose moiety or modified
pentose moiety attached to a heterocyclic base or tautomer thereof
via a N-glycosidic bond, such as attached via the 9-position of a
purine-base or the 1-position of a pyrimidine-base. Examples
include, but are not limited to, a ribonucleoside comprising a
ribose moiety and a deoxyribonucleoside comprising a deoxyribose
moiety. A modified pentose moiety is a pentose moiety in which an
oxygen atom has been replaced with a carbon and/or a carbon has
been replaced with a sulfur or an oxygen atom. A "nucleoside" is a
monomer that can have a substituted base and/or sugar moiety.
Additionally, a nucleoside can be incorporated into larger DNA
and/or RNA polymers and oligomers. In some instances, the
nucleoside can be a nucleoside analog drug.
[0067] The term "nucleotide" is used herein in its ordinary sense
as understood by those skilled in the art, and refers to a
nucleoside having a phosphate ester bound to the pentose moiety,
for example, at the 5'-position.
[0068] As used herein, the term "heterocyclic base" refers to an
optionally substituted nitrogen-containing heterocyclyl that can be
attached to an optionally substituted pentose moiety or modified
pentose moiety. In some embodiments, the heterocyclic base can be
selected from an optionally substituted purine-base, an optionally
substituted pyrimidine-base and an optionally substituted
triazole-base (for example, a 1,2,4-triazole). The term
"purine-base" is used herein in its ordinary sense as understood by
those skilled in the art, and includes its tautomers. Similarly,
the term "pyrimidine-base" is used herein in its ordinary sense as
understood by those skilled in the art, and includes its tautomers.
A non-limiting list of optionally substituted purine-bases includes
purine, adenine, guanine, hypoxanthine, xanthine, alloxanthine,
7-alkylguanine (e.g. 7-methylguanine), theobromine, caffeine, uric
acid and isoguanine. Examples of pyrimidine-bases include, but are
not limited to, cytosine, thymine, uracil, 5,6-dihydrouracil and
5-alkylcytosine (e.g., 5-methylcytosine). An example of an
optionally substituted triazole-base is
1,2,4-triazole-3-carboxamide. Other non-limiting examples of
heterocyclic bases include diaminopurine,
8-oxo-N.sup.6-alkyladenine (e.g., 8-oxo-N.sup.6-methyladenine),
7-deazaxanthine, 7-deazaguanine, 7-deazaadenine,
N.sup.4,N.sup.4-ethanocytosin,
N.sup.6,N.sup.6-ethano-2,6-diaminopurine, 5-halouracil (e.g.,
5-fluorouracil and 5-bromouracil), pseudoisocytosine, isocytosine,
isoguanine, and other heterocyclic bases described in U.S. Pat.
Nos. 5,432,272 and 7,125,855, which are incorporated herein by
reference for the limited purpose of disclosing additional
heterocyclic bases. In some embodiments, a heterocyclic base can be
optionally substituted with an amine or an enol protecting
group(s).
[0069] The term "--N-linked amino acid" refers to an amino acid
that is attached to the indicated moiety via a main-chain amino or
mono-substituted amino group. When the amino acid is attached in an
--N-linked amino acid, one of the hydrogens that is part of the
main-chain amino or mono-substituted amino group is not present and
the amino acid is attached via the nitrogen. N-linked amino acids
can be substituted or unsubstituted.
[0070] The term "--N-linked amino acid ester derivative" refers to
an amino acid in which a main-chain carboxylic acid group has been
converted to an ester group. In some embodiments, the ester group
has a formula selected from alkyl-O--C(.dbd.O)--,
cycloalkyl-O--C(.dbd.O)--, aryl-O--C(.dbd.O)-- and
aryl(alkyl)-O--C(.dbd.O)--. A non-limiting list of ester groups
include substituted and unsubstituted versions of the following:
methyl-O--C(.dbd.O)--, ethyl-O--C(.dbd.O)--,
n-propyl-O--C(.dbd.O)--, isopropyl-O--C(.dbd.O)--,
n-butyl-O--C(.dbd.O)--, isobutyl-O--C(.dbd.O)--,
tert-butyl-O--C(.dbd.O)--, neopentyl-O--C(.dbd.O)--,
cyclopropyl-O--C(.dbd.O)--, cyclobutyl-O--C(.dbd.O)--,
cyclopentyl-O--C(.dbd.O)--, cyclohexyl-O--C(.dbd.O)--,
phenyl-O--C(.dbd.O)--, benzyl-O--C(.dbd.O)--, and
naphthyl-O--C(.dbd.O)--. N-linked amino acid ester derivatives can
be substituted or unsubstituted.
[0071] The term "--O-linked amino acid" refers to an amino acid
that is attached to the indicated moiety via the hydroxy from its
main-chain carboxylic acid group. When the amino acid is attached
in an --O-linked amino acid, the hydrogen that is part of the
hydroxy from its main-chain carboxylic acid group is not present
and the amino acid is attached via the oxygen. O-linked amino acids
can be substituted or unsubstituted.
[0072] As used herein, the term "amino acid" refers to any amino
acid (both standard and non-standard amino acids), including, but
not limited to, .alpha.-amino acids, .beta.-amino acids,
.gamma.-amino acids and .delta.-amino acids. Examples of suitable
amino acids include, but are not limited to, alanine, asparagine,
aspartate, cysteine, glutamate, glutamine, glycine, proline,
serine, tyrosine, arginine, histidine, isoleucine, leucine, lysine,
methionine, phenylalanine, threonine, tryptophan and valine.
Additional examples of suitable amino acids include, but are not
limited to, ornithine, hypusine, 2-aminoisobutyric acid,
dehydroalanine, gamma-aminobutyric acid, citrulline, beta-alanine,
alpha-ethyl-glycine, alpha-propyl-glycine and norleucine.
[0073] The terms "phosphorothioate" and "phosphothioate" refer to a
compound of the general formula
##STR00002##
its protonated forms (for example,
##STR00003##
and its tautomers (such as
##STR00004##
[0074] As used herein, the term "phosphate" is used in its ordinary
sense as understood by those skilled in the art, and includes its
protonated forms (for example,
##STR00005##
As used herein, the terms "monophosphate," "diphosphate," and
"triphosphate" are used in their ordinary sense as understood by
those skilled in the art, and include protonated forms.
[0075] The terms "protecting group" and "protecting groups" as used
herein refer to any atom or group of atoms that is added to a
molecule in order to prevent existing groups in the molecule from
undergoing unwanted chemical reactions. Examples of protecting
group moieties are described in T. W. Greene and P. G. M. Wuts,
Protective Groups in Organic Synthesis, 3. Ed. John Wiley &
Sons, 1999, and in J. F. W. McOmie, Protective Groups in Organic
Chemistry Plenum Press, 1973, both of which are hereby incorporated
by reference for the limited purpose of disclosing suitable
protecting groups. The protecting group moiety may be chosen in
such a way, that they are stable to certain reaction conditions and
readily removed at a convenient stage using methodology known from
the art. A non-limiting list of protecting groups include benzyl;
substituted benzyl; alkylcarbonyls and alkoxycarbonyls (e.g.,
t-butoxycarbonyl (BOC), acetyl, or isobutyryl); arylalkylcarbonyls
and arylalkoxycarbonyls (e.g., benzyloxycarbonyl); substituted
methyl ether (e.g. methoxymethyl ether); substituted ethyl ether; a
substituted benzyl ether; tetrahydropyranyl ether, silyls (e.g.,
trimethylsilyl, triethylsilyl, triisopropylsilyl,
t-butyldimethylsilyl, tri-iso-propylsilyloxymethyl,
[2-(trimethylsilyl)ethoxy]methyl or t-butyldiphenylsilyl); esters
(e.g. benzoate ester); carbonates (e.g. methoxymethylcarbonate);
sulfonates (e.g. tosylate or mesylate); acyclic ketal (e.g.
dimethyl acetal); cyclic ketals (e.g., 1,3-dioxane, 1,3-dioxolanes,
and those described herein); acyclic acetal; cyclic acetal (e.g.,
those described herein); acyclic hemiacetal; cyclic hemiacetal;
cyclic dithioketals (e.g., 1,3-dithiane or 1,3-dithiolane);
orthoesters (e.g., those described herein) and triarylmethyl groups
(e.g., trityl; monomethoxytrityl (MMTr); 4,4'-dimethoxytrityl
(DMTr); 4,4',4''-trimethoxytrityl (TMTr); and those described
herein).
[0076] The term "pharmaceutically acceptable salt" refers to a salt
of a compound that does not cause significant irritation to an
organism to which it is administered and does not abrogate the
biological activity and properties of the compound. In some
embodiments, the salt is an acid addition salt of the compound.
Pharmaceutical salts can be obtained by reacting a compound with
inorganic acids such as hydrohalic acid (e.g., hydrochloric acid or
hydrobromic acid), sulfuric acid, nitric acid and phosphoric acid.
Pharmaceutical salts can also be obtained by reacting a compound
with an organic acid such as aliphatic or aromatic carboxylic or
sulfonic acids, for example formic, acetic, succinic, lactic,
malic, tartaric, citric, ascorbic, nicotinic, methanesulfonic,
ethanesulfonic, p-toluenesulfonic, salicylic or naphthalenesulfonic
acid. Pharmaceutical salts can also be obtained by reacting a
compound with a base to form a salt such as an ammonium salt, an
alkali metal salt, such as a sodium or a potassium salt, an
alkaline earth metal salt, such as a calcium or a magnesium salt, a
salt of organic bases such as dicyclohexylamine,
N-methyl-D-glucamine, tris(hydroxymethyl)methylamine,
C.sub.1-C.sub.7 alkylamine, cyclohexylamine, triethanolamine,
ethylenediamine, and salts with amino acids such as arginine and
lysine.
[0077] Terms and phrases used in this application, and variations
thereof, especially in the appended claims, unless otherwise
expressly stated, should be construed as open ended as opposed to
limiting. As examples of the foregoing, the term `including` should
be read to mean `including, without limitation,` `including but not
limited to,` or the like; the term `comprising` as used herein is
synonymous with `including,` `containing,` or `characterized by,`
and is inclusive or open-ended and does not exclude additional,
unrecited elements or method steps; the term `having` should be
interpreted as `having at least;` the term `includes` should be
interpreted as `includes but is not limited to;` the term `example`
is used to provide exemplary instances of the item in discussion,
not an exhaustive or limiting list thereof; and use of terms like
`preferably,` `preferred,` `desired,` or `desirable,` and words of
similar meaning should not be understood as implying that certain
features are critical, essential, or even important to the
structure or function of the invention, but instead as merely
intended to highlight alternative or additional features that may
or may not be utilized in a particular embodiment. In addition, the
term "comprising" is to be interpreted synonymously with the
phrases "having at least" or "including at least". When used in the
context of a process, the term "comprising" means that the process
includes at least the recited steps, but may include additional
steps. When used in the context of a compound, composition or
device, the term "comprising" means that the compound, composition
or device includes at least the recited features or components, but
may also include additional features or components. Likewise, a
group of items linked with the conjunction `and` should not be read
as requiring that each and every one of those items be present in
the grouping, but rather should be read as `and/or` unless
expressly stated otherwise. Similarly, a group of items linked with
the conjunction `or` should not be read as requiring mutual
exclusivity among that group, but rather should be read as `and/or`
unless expressly stated otherwise.
[0078] With respect to the use of substantially any plural and/or
singular terms herein, those having skill in the art can translate
from the plural to the singular and/or from the singular to the
plural as is appropriate to the context and/or application. The
various singular/plural permutations may be expressly set forth
herein for sake of clarity. The indefinite article "a" or "an" does
not exclude a plurality. A single processor or other unit may
fulfill the functions of several items recited in the claims. The
mere fact that certain measures are recited in mutually different
dependent claims does not indicate that a combination of these
measures cannot be used to advantage. Any reference signs in the
claims should not be construed as limiting the scope.
[0079] It is understood that, in any compound described herein
having one or more chiral centers, if an absolute stereochemistry
is not expressly indicated, then each center may independently be
of R-configuration or S-configuration or a mixture thereof. Thus,
the compounds provided herein may be enantiomerically pure,
enantiomerically enriched, racemic mixture, diastereomerically
pure, diastereomerically enriched, or a stereoisomeric mixture. In
addition it is understood that, in any compound described herein
having one or more double bond(s) generating geometrical isomers
that can be defined as E or Z, each double bond may independently
be E or Z a mixture thereof.
[0080] Likewise, it is understood that, in any compound described,
all tautomeric forms are also intended to be included. For example
all tautomers of a phosphate and a phosphorothioate groups are
intended to be included. Examples of tautomers of a
phosphorothioate include the following:
##STR00006##
Furthermore, all tautomers of heterocyclic bases known in the art
are intended to be included, including tautomers of natural and
non-natural purine-bases and pyrimidine-bases.
[0081] It is to be understood that where compounds disclosed herein
have unfilled valencies, then the valencies are to be filled with
hydrogens or isotopes thereof, e.g., hydrogen-1 (protium) and
hydrogen-2 (deuterium).
[0082] It is understood that the compounds described herein can be
labeled isotopically. Substitution with isotopes such as deuterium
may afford certain therapeutic advantages resulting from greater
metabolic stability, such as, for example, increased in vivo
half-life or reduced dosage requirements. Each chemical element as
represented in a compound structure may include any isotope of said
element. For example, in a compound structure a hydrogen atom may
be explicitly disclosed or understood to be present in the
compound. At any position of the compound that a hydrogen atom may
be present, the hydrogen atom can be any isotope of hydrogen,
including but not limited to hydrogen-1 (protium) and hydrogen-2
(deuterium). Thus, reference herein to a compound encompasses all
potential isotopic forms unless the context clearly dictates
otherwise.
[0083] It is understood that the methods and combinations described
herein include crystalline forms (also known as polymorphs, which
include the different crystal packing arrangements of the same
elemental composition of a compound), amorphous phases, salts,
solvates, and hydrates. In some embodiments, the compounds
described herein exist in solvated forms with pharmaceutically
acceptable solvents such as water, ethanol, or the like. In other
embodiments, the compounds described herein exist in unsolvated
form. Solvates contain either stoichiometric or non-stoichiometric
amounts of a solvent, and may be formed during the process of
crystallization with pharmaceutically acceptable solvents such as
water, ethanol, or the like. Hydrates are formed when the solvent
is water, or alcoholates are formed when the solvent is alcohol. In
addition, the compounds provided herein can exist in unsolvated as
well as solvated forms. In general, the solvated forms are
considered equivalent to the unsolvated forms for the purposes of
the compounds and methods provided herein.
[0084] Where a range of values is provided, it is understood that
the upper and lower limit, and each intervening value between the
upper and lower limit of the range is encompassed within the
embodiments.
Compounds
[0085] Some embodiments disclosed herein relate to a compound
selected from Formula (I), Formula (II) and Formula (III), or a
pharmaceutically acceptable salt of the foregoing:
##STR00007##
wherein: B.sup.1A, B.sup.1B and B.sup.1C can be independently an
optionally substituted heterocyclic base or an optionally
substituted heterocyclic base with a protected amino group; RIA can
be selected from hydrogen, an optionally substituted acyl, an
optionally substituted O-linked amino acid,
##STR00008##
when the dashed line () of Formula (I) is a single bond, R.sup.2A
can be CH.sub.2, and R.sup.3A can be O (oxygen); when the dashed
line () of Formula (I) is absent, R.sup.2A can be selected from an
optionally substituted C.sub.1-6 alkyl, an optionally substituted
C.sub.2-6 alkenyl, an optionally substituted C.sub.2-6 alkynyl, an
optionally substituted C.sub.3-6 cycloalkyl, an optionally
substituted --O--C.sub.1-6 alkyl, an optionally substituted
O--C.sub.3-6 alkenyl, an optionally substituted O--C.sub.3-6
alkenyl and cyano, and R.sup.3A can be selected from OH,
--OC(.dbd.O)R.sup.''A and an optionally substituted O-linked amino
acid; R.sup.1B can be selected from O.sup.-, OH,
##STR00009##
an optionally substituted N-linked amino acid and an optionally
substituted N-linked amino acid ester derivative; R.sup.1C and
R.sup.2C can be independently selected from O.sup.-, OH, an
optionally substituted C.sub.1-6 alkoxy,
##STR00010##
optionally substituted N-linked amino acid and an optionally
substituted N-linked amino acid ester derivative; or R.sup.1C can
be
##STR00011##
and R.sup.2C can be O.sup.- or OH; R.sup.2B and R.sup.3C can be
independently selected from an optionally substituted C.sub.1-6
alkyl, an optionally substituted C.sub.2-6 alkenyl, an optionally
substituted C.sub.2-6 alkynyl, an optionally substituted
--O--C.sub.1-6 alkyl, an optionally substituted --O--C.sub.3-6
alkenyl, an optionally substituted-O--C.sub.3-6 alkynyl, an
optionally substituted C.sub.3-6 cycloalkyl and cyano; R.sup.4C can
be selected from OH, --OC(.dbd.O)R.sup.''C and an optionally
substituted O-linked amino acid; R.sup.4A, R.sup.3B and R.sup.5C
can be independently a halogen; R.sup.5A, R.sup.4B and R.sup.6C can
be independently hydrogen or halogen; R.sup.6A, R.sup.7A and
R.sup.8A can be independently selected from absent, hydrogen, an
optionally substituted C.sub.1-24 alkyl, an optionally substituted
C.sub.2-24 alkenyl, an optionally substituted C.sub.2-24 alkynyl,
an optionally substituted C.sub.3-6 cycloalkyl, an optionally
substituted C.sub.3-6 cycloalkenyl, an optionally substituted aryl,
an optionally substituted heteroaryl, an optionally substituted
aryl(C.sub.1-6 alkyl), an optionally substituted
*--(CR.sup.15AR.sup.16A).sub.p--O--C.sub.1-24 alkyl, an optionally
substituted *--(CR.sup.17AR.sup.18A).sub.q--C.sub.1-24 alkenyl,
##STR00012##
or R.sup.6A can be
##STR00013##
[0086] and R.sup.7A can be absent or hydrogen; or R.sup.6A and
R.sup.7A can be taken together to form a moiety selected from an
optionally substituted
##STR00014##
and an optionally substituted
##STR00015##
wherein the oxygens connected to R.sup.6A and R.sup.7A, the
phosphorus and the moiety form a six-membered to ten-membered ring
system; R.sup.9A can be independently selected from an optionally
substituted C.sub.1-24 alkyl, an optionally substituted C.sub.2-24
alkenyl, an optionally substituted C.sub.2-24 alkynyl, an
optionally substituted C.sub.3-6 cycloalkyl, an optionally
substituted C.sub.3-6 cycloalkenyl, NR.sup.30AR.sup.31A, an
optionally substituted N-linked amino acid and an optionally
substituted N-linked amino acid ester derivative; R.sup.10A and
R.sup.11A can be independently an optionally substituted N-linked
amino acid or an optionally substituted N-linked amino acid ester
derivative; R.sup.12A, R.sup.13A and R.sup.14A can be independently
absent or hydrogen; each R.sup.15A, each R.sup.16A, each R.sup.17A
and each R.sup.18A can be independently hydrogen, an optionally
substituted C.sub.1-24 alkyl or alkoxy; R.sup.19A, R.sup.20A,
R.sup.22A, R.sup.23A, R.sup.5B, R.sup.6B, R.sup.8B, R.sup.9B,
R.sup.9C, R.sup.10C, R.sup.12C and R.sup.13C can be independently
selected from hydrogen, an optionally substituted C.sub.1-24 alkyl
and an optionally substituted aryl; R.sup.21A, R.sup.24A, R.sup.7B,
R.sup.10B, R.sup.11C and R.sup.14C can be independently selected
from hydrogen, an optionally substituted C.sub.1-24 alkyl, an
optionally substituted aryl, an optionally substituted
--O--C.sub.1-24 alkyl and an optionally substituted --O-aryl;
R.sup.25A, R.sup.29A, R.sup.11B and R.sup.15C can be independently
selected from hydrogen, an optionally substituted C.sub.1-24 alkyl
and an optionally substituted aryl; R.sup.16C, R.sup.17C and
R.sup.18C can be independently absent or hydrogen; R.sup.26A and
R.sup.27A can be independently --C.ident.N or an optionally
substituted substituent selected from C.sub.2-8 organylcarbonyl,
C.sub.2-8 alkoxycarbonyl and C.sub.2-8 organylaminocarbonyl;
R.sup.28A can be selected from hydrogen, an optionally substituted
C.sub.1-24-alkyl, an optionally substituted C.sub.2-24 alkenyl, an
optionally substituted C.sub.2-24 alkynyl, an optionally
substituted C.sub.3-6 cycloalkyl and an optionally substituted
C.sub.3-6 cycloalkenyl; R.sup.30A and R.sup.31A can be
independently selected from hydrogen, an optionally substituted
C.sub.1-24-alkyl, an optionally substituted C.sub.2-24 alkenyl, an
optionally substituted C.sub.2-24 alkynyl, an optionally
substituted C.sub.3-6 cycloalkyl and an optionally substituted
C.sub.3-6 cycloalkenyl; for Formula (III), can be a single bond or
a double bond; when is a single bond, each R.sup.7C and each
R.sup.8C can be independently hydrogen or halogen; and when is a
double bond, each R.sup.7C is absent and each R.sup.8C can be
independently hydrogen or halogen; R.sup.''A and R.sup.''C can be
independently an optionally substituted C.sub.1-24-alkyl, m and n
can be independently 0 or 1; p and q can be independently selected
from 1, 2 and 3; r can be 1 or 2; Z.sup.1A, Z.sup.2A, Z.sup.3A,
Z.sup.4A, Z.sup.1B, Z.sup.2B and Z.sup.1C can be independently O or
S; and provided that when the dashed line () of Formula (I) is
absent; R.sup.1A is
##STR00016##
wherein R.sup.8A is an unsubstituted C.sub.1-4 alkyl or phenyl
optionally para-substituted with a halogen or methyl and R.sup.9A
is methyl ester, ethyl ester, isopropyl ester, n-butyl ester,
benzyl ester or phenyl ester of an amino acid selected from
glycine, alanine, valine, leucine, phenylalanine, tryptophan,
methionine and proline; R.sup.3A is OH; R.sup.4A is fluoro;
R.sup.5A is fluoro or hydrogen; and B.sup.1A is an unsubstituted
uracil; then R.sup.2A cannot be --OCH.sub.3; provided that when the
dashed line () of Formula (I) is absent; R.sup.1A is H; R.sup.3A is
OH; R.sup.4A is fluoro; R.sup.5A is fluoro; and B.sup.1A is an
unsubstituted cytosine; then R.sup.2A cannot be allenyl; provided
that when the dashed line () of Formula (I) is absent; R.sup.1A is
H; R.sup.3A is OH; R.sup.4A is fluoro; R.sup.5A is hydrogen; and
B.sup.1A is an unsubstituted thymine; then R.sup.2A cannot be
C.sub.1 alkyl substituted with an optionally substituted N-amido
(for example, --NC(.dbd.O)CF.sub.3); and provided that when the
dashed line () of Formula (I) is absent; R.sup.1A is H; R.sup.3A is
OH; R.sup.4A is fluoro; R.sup.5A is fluoro; and B.sup.1A is an
unsubstituted cytosine; then R.sup.2A cannot be ethynyl.
[0087] In some embodiments, the compound can be a compound of
Formula (I), or a pharmaceutically acceptable salt thereof,
wherein: B.sup.1A can be an optionally substituted heterocyclic
base or an optionally substituted heterocyclic base with a
protected amino group; R.sup.1A can be selected from hydrogen,
##STR00017##
when the dashed line () of Formula (I) is a single bond, R.sup.2A
is CH.sub.2, and R.sup.3A is O (oxygen); when the dashed line () of
Formula (I) is absent, R.sup.2A can be selected from an optionally
substituted C.sub.1-6 alkyl, an optionally substituted C.sub.2-6
alkenyl, an optionally substituted C.sub.2-6 alkynyl, an optionally
substituted --O--C.sub.1-6 alkyl, an optionally substituted
--O--C.sub.3-6 alkenyl, an optionally substituted --O--C.sub.3-6
alkynyl and cyano, and R.sup.3A is OH; R.sup.4A can be a halogen;
R.sup.5A can be hydrogen or halogen; R.sup.6A, R.sup.7A and
R.sup.8A can be independently selected from absent, hydrogen, an
optionally substituted C.sub.1-24 alkyl, an optionally substituted
C.sub.2-24 alkenyl, an optionally substituted C.sub.2-24 alkynyl,
an optionally substituted C.sub.3-6 cycloalkyl, an optionally
substituted C.sub.3-6 cycloalkenyl, an optionally substituted aryl,
an optionally substituted heteroaryl, an optionally substituted
aryl(C.sub.1-6 alkyl), an optionally substituted
*--(CR.sup.15AR.sup.16A).sub.p--O--C.sub.1-24 alkyl, an optionally
substituted *--(CR.sup.17AR.sup.18A).sub.q--O--C.sub.1-24
alkenyl,
##STR00018##
or R.sup.6A can be
##STR00019##
[0088] and R.sup.7A can be absent or hydrogen; or R.sup.6A and
R.sup.7A can be taken together to form a moiety selected from an
optionally substituted
##STR00020##
and an optionally substituted
##STR00021##
wherein the oxygens connected to R.sup.6A and R.sup.7A, the
phosphorus and the moiety form a six-membered to ten-membered ring
system. R.sup.9A can be independently selected from an optionally
substituted C.sub.1-24 alkyl, an optionally substituted C.sub.2-24
alkenyl, an optionally substituted C.sub.2-24 alkynyl, an
optionally substituted C.sub.3-6 cycloalkyl, an optionally
substituted C.sub.3-6 cycloalkenyl, NR.sup.30AR.sup.31A, an
optionally substituted N-linked amino acid and an optionally
substituted N-linked amino acid ester derivative; R.sup.10A and
R.sup.11A can be independently an optionally substituted N-linked
amino acid or an optionally substituted N-linked amino acid ester
derivative; R.sup.1A, R.sup.13A and R.sup.14A can be independently
absent or hydrogen; each R.sup.15A, each R.sup.16A, each R.sup.17A
and each R.sup.18A can be independently hydrogen, an optionally
substituted C.sub.1-24 alkyl or alkoxy; R.sup.19A, R.sup.20A,
R.sup.22A and R.sup.23A can be independently selected from
hydrogen, an optionally substituted C.sub.1-24 alkyl and an
optionally substituted aryl; R.sup.21A and R.sup.24A can be
independently selected from hydrogen, an optionally substituted
C.sub.1-24 alkyl, an optionally substituted aryl, an optionally
substituted --O--C.sub.1-24 alkyl and an optionally substituted
--O-aryl; R.sup.25A and R.sup.29A can be independently selected
from hydrogen, an optionally substituted C.sub.1-24 alkyl and an
optionally substituted aryl; R.sup.26A and R.sup.27A can be
independently --C.ident.N or an optionally substituted substituent
selected from C.sub.2-8 organylcarbonyl, C.sub.2-8 alkoxycarbonyl
and C.sub.2-8 organylaminocarbonyl; R.sup.28A can be selected from
hydrogen, an optionally substituted C.sub.1-24-alkyl, an optionally
substituted C.sub.2-24 alkenyl, an optionally substituted
C.sub.2-24 alkynyl, an optionally substituted C.sub.3-6 cycloalkyl
and an optionally substituted C.sub.3-6 cycloalkenyl; R.sup.30A and
R.sup.31A can be independently selected from hydrogen, an
optionally substituted C.sub.1-24-alkyl, an optionally substituted
C.sub.2-24 alkenyl, an optionally substituted C.sub.2-24 alkynyl,
an optionally substituted C.sub.3-6 cycloalkyl and an optionally
substituted C.sub.3-6 cycloalkenyl; m can be 0 or 1; p and q can be
independently selected from 1, 2 and 3; r can be 1 or 2; Z.sup.1A,
Z.sup.2A, Z.sup.3A and Z.sup.4A can be independently O or S. In
some embodiments, a compound of Formula (I) can have a structure
shown herein, provided that when the dashed line () of Formula (I)
is absent; R.sup.1A is
##STR00022##
wherein R.sup.8A is an unsubstituted C.sub.1-4 alkyl or phenyl
optionally para-substituted with a halogen or methyl and R.sup.9A
is methyl ester, ethyl ester, isopropyl ester, n-butyl ester,
benzyl ester or phenyl ester of an amino acid selected from
glycine, alanine, valine, leucine, phenylalanine, tryptophan,
methionine and proline; R.sup.3A is OH; R.sup.4A is fluoro;
R.sup.5A is fluoro or hydrogen; and B.sup.1A is an unsubstituted
uracil; then R.sup.2A cannot be --OCH.sub.3; provided that when the
dashed line () of Formula (I) is absent; R.sup.1A is H; R.sup.3A is
OH; R.sup.4A is fluoro; R.sup.5A is fluoro; and B.sup.1A is an
unsubstituted cytosine; then R.sup.2A cannot be allenyl; provided
that when the dashed line () of Formula (I) is absent; R.sup.1A is
H; R.sup.3A is OH; R.sup.4A is fluoro; R.sup.5A is hydrogen; and
B.sup.1A is an unsubstituted thymine; then R.sup.2A cannot be C
alkyl substituted with an N-amido; and provided that when the
dashed line () of Formula (I) is absent; R.sup.1A is H; R.sup.3A is
OH; R.sup.4A is fluoro; R.sup.5A is fluoro; and B.sup.1A is an
unsubstituted cytosine; then R.sup.2A cannot be ethynyl.
[0089] In some embodiments, R.sup.1A can be
##STR00023##
In some embodiments, R.sup.6A and R.sup.7A can be both hydrogen. In
other embodiments, R.sup.6A and R.sup.7A can be both absent. In
still other embodiments, at least one R.sup.6A and R.sup.7A can be
absent. In yet still other embodiments, at least one R.sup.6A and
R.sup.7A can be hydrogen. Those skilled in the art understand that
when R.sup.6A and/or R.sup.7A are absent, the associated oxygen(s)
will have a negative charge. For example, when R.sup.6A is absent,
the oxygen associated with R.sup.6A will have a negative charge. In
some embodiments, Z.sup.1A can be O (oxygen). In other embodiments,
Z.sup.1A can be S (sulfur). In some embodiments, R.sup.1A can be a
monophosphate. In other embodiments, R.sup.1A can be a
monothiophosphate.
[0090] In some embodiments, when R.sup.1A is
##STR00024##
one of R.sup.6A and R.sup.7A can be hydrogen, and the other of
R.sup.6A and R.sup.7A is selected from an optionally substituted
C.sub.1-24 alkyl, an optionally substituted C.sub.2-24 alkenyl, an
optionally substituted C.sub.2-24 alkynyl, an optionally
substituted C.sub.3-6 cycloalkyl, an optionally substituted
C.sub.3-6 cycloalkenyl, an optionally substituted aryl, an
optionally substituted heteroaryl and an optionally substituted
aryl(C.sub.1-6 alkyl). In some embodiments, one of R.sup.6A and
R.sup.7A can be hydrogen, and the other of R.sup.6A and R.sup.7A
can be an optionally substituted C.sub.1-24 alkyl. In other
embodiments, both R.sup.6A and R.sup.7A can be independently
selected from an optionally substituted C.sub.1-24 alkyl, an
optionally substituted C.sub.2-24 alkenyl, an optionally
substituted C.sub.2-24 alkynyl, an optionally substituted C.sub.3.
6 cycloalkyl, an optionally substituted C.sub.3-6 cycloalkenyl, an
optionally substituted aryl, an optionally substituted heteroaryl
and an optionally substituted aryl(C.sub.1-6 alkyl). In some
embodiments, both R.sup.6A and R.sup.7A can be an optionally
substituted C.sub.1-24 alkyl. In other embodiments, both R.sup.6A
and R.sup.7A can be an optionally substituted C.sub.2-24 alkenyl.
In some embodiments, R.sup.6A and R.sup.7A can be independently an
optionally substituted version of the following: myristoleyl,
myristyl, palmitoleyl, palmityl, sapienyl, oleyl, elaidyl,
vaccenyl, linoleyl, .alpha.-linolenyl, arachidonyl,
eicosapentaenyl, erucyl, docosahexaenyl, caprylyl, capryl, lauryl,
stearyl, arachidyl, behenyl, lignoceryl, and cerotyl.
[0091] In some embodiments, at least one of R.sup.6A and R.sup.7A
can be *--(CR.sup.15AR.sup.16A).sub.p--O--C.sub.1-24 alkyl. In
other embodiments, R.sup.6A and R.sup.7A can be both
*--(CR.sup.15AR.sup.16Ap C.sub.1-24 alkyl. In some embodiments,
each R.sup.15A and each R.sup.16A are hydrogen. In other
embodiments, at least one of R.sup.15A and R.sup.16A is an
optionally substituted C.sub.1-24 alkyl. In other embodiments, at
least one of R.sup.15A and R.sup.16A is an alkoxy (for example,
benzoxy). In some embodiments, p can be 1. In other embodiments, p
can be 2. In still other embodiments, p can be 3.
[0092] In some embodiments, at least one of R.sup.6A and R.sup.7A
can be *--(CR.sup.17AR.sup.18A).sub.q--O--C.sub.2-24 alkenyl. In
other embodiments, R.sup.6A and R.sup.7A can be both
*--(CR.sup.17AR.sup.18A).sub.q2-24 alkenyl. In some embodiments,
each R.sup.17A and each R.sup.18A are hydrogen. In other
embodiments, at least one of R.sup.17A and R.sup.18A is an
optionally substituted C.sub.1-24 alkyl. In some embodiments, q can
be 1. In other embodiments, q can be 2. In still other embodiments,
q can be 3. When at least one of R.sup.6A and R.sup.7A is
*--(CR.sup.15AR.sup.16A).sub.p--O--C.sub.1-24 alkyl or
*--(CR.sup.17AR.sup.18A).sub.q--O--C.sub.2-24 alkenyl, the
C.sub.1-24 alkyl can be selected from caprylyl, capryl, lauryl,
myristyl, palmityl, stearyl, arachidyl, behenyl, lignoceryl, and
cerotyl, and the C.sub.2-24 alkenyl can be selected from
myristoleyl, palmitoleyl, sapienyl, oleyl, elaidyl, vaccenyl,
linoleyl, .alpha.-linolenyl, arachidonyl, eicosapentaenyl, erucyl
and docosahexaenyl.
[0093] In some embodiments, when R.sup.1A is
##STR00025##
at least one of R.sup.6A and R.sup.7A can be selected from
##STR00026##
and the other of R.sup.6A and R.sup.7A can be selected from absent,
hydrogen, an optionally substituted C.sub.1-24 alkyl, an optionally
substituted C.sub.2-24 alkenyl, an optionally substituted
C.sub.2-24 alkynyl, an optionally substituted C.sub.3-6 cycloalkyl,
an optionally substituted C.sub.3-6 cycloalkenyl, an optionally
substituted aryl, an optionally substituted heteroaryl and an
optionally substituted aryl(C.sub.1-6 alkyl).
[0094] In some embodiments, at least one of R.sup.6A and R.sup.7A
can be
##STR00027##
In some embodiments, both R.sup.6A and R.sup.7A can be
##STR00028##
When one or both of R.sup.6A and R.sup.7A are
##STR00029##
R.sup.19A and R.sup.20A can be independently selected from
hydrogen, an optionally substituted C.sub.1-24 alkyl and an
optionally substituted aryl; and R.sup.21A can be selected from
hydrogen, an optionally substituted C.sub.1-24 alkyl, an optionally
substituted aryl, an optionally substituted --O--C.sub.1-24 alkyl
and an optionally substituted --O-aryl. In some embodiments,
R.sup.19A and R.sup.20A can be hydrogen. In other embodiments, at
least one of R.sup.19A and R.sup.20A can be an optionally
substituted C.sub.1-24 alkyl or an optionally substituted aryl. In
some embodiments, R.sup.21A can be an optionally substituted
C.sub.1-24 alkyl. In other embodiments, R.sup.21A can be an
optionally substituted aryl. In still other embodiments, R.sup.21A
can be an optionally substituted --O--C.sub.1-24 alkyl or an
optionally substituted --O-aryl.
[0095] In some embodiments, both R.sup.6A and R.sup.7A can be
##STR00030##
When one or both of R.sup.6A and R.sup.7A are
##STR00031##
R.sup.22A and R.sup.23A can be independently selected from
hydrogen, an optionally substituted C.sub.1-24 alkyl and an
optionally substituted aryl; R.sup.24A can be independently
selected from hydrogen, an optionally substituted C.sub.1-24 alkyl,
an optionally substituted aryl, an optionally substituted
--C.sub.1-24 alkyl and an optionally substituted --O-aryl; and
Z.sup.4A can be independently O (oxygen) or S (sulfur). In some
embodiments, R.sup.22A and R.sup.23A can be hydrogen. In other
embodiments, at least one of R.sup.22A and R.sup.23A can be an
optionally substituted C.sub.1-24 alkyl or an optionally
substituted aryl. In some embodiments, R.sup.24A can be an
optionally substituted C.sub.1-24 alkyl. In other embodiments,
R.sup.24A can be an optionally substituted aryl. In still other
embodiments, R.sup.24A can be an optionally substituted
--O--C.sub.1-24 alkyl or an optionally substituted --O-aryl. In
some embodiments, Z.sup.4A can be O (oxygen). In other embodiments,
Z.sup.4A can be or S (sulfur). In some embodiments, one or both of
R.sup.6A and R.sup.7A can be isopropylcarbonyloxymethyl. In some
embodiments, one or both of R.sup.6A and R.sup.7A can be
pivaloyloxymethyl.
[0096] In some embodiments, both R.sup.6A and R.sup.7A can be
##STR00032##
When one or both of R.sup.6A and R.sup.7A are
##STR00033##
R.sup.26A and R.sup.27A can be independently --C.ident.N or an
optionally substituted substituent selected from C.sub.2-8
organylcarbonyl, C.sub.2-4 alkoxycarbonyl and C.sub.2-8
organylaminocarbonyl; R.sup.2A can be selected from hydrogen, an
optionally substituted C.sub.1-24-alkyl, an optionally substituted
C.sub.2-24 alkenyl, an optionally substituted C.sub.2-24 alkynyl,
an optionally substituted C.sub.3-6 cycloalkyl and an optionally
substituted C.sub.3-6 cycloalkenyl; and r can be 1 or 2. In some
embodiments, R.sup.26A can be --C.ident.N and R.sup.27A can be an
optionally substituted C.sub.2-8 alkoxycarbonyl, such as
--C(.dbd.O)OCH.sub.3. In other embodiments, R.sup.26A can be
--C.ident.N and R.sup.27A can be an optionally substituted
C.sub.2-8 organylaminocarbonyl, for example,
--C(.dbd.O)NHCH.sub.2CH.sub.3 and
--C(.dbd.O)NHCH.sub.2CH.sub.2phenyl. In some embodiments, both
R.sup.26A and R.sup.27A can be an optionally substituted C.sub.2-8
organylcarbonyl, such as --C(.dbd.O)CH.sub.3. In some embodiments,
both R.sup.26A and R.sup.27A can be an optionally substituted
C.sub.1-8 alkoxycarbonyl, for example, --C(.dbd.O)OCH.sub.2CH.sub.3
and --C(.dbd.O)OCH.sub.3. In some embodiments, including those
described in this paragraph, R.sup.28A can be an optionally
substituted C.sub.1-4-alkyl. In some embodiment, R.sup.28A can be
methyl or tert-butyl. In some embodiments, r can be 1. In other
embodiments, r can be 2.
[0097] Example of
##STR00034##
include, but are not limited to the following:
##STR00035##
[0098] In some embodiments, R.sup.6A and R.sup.7A can be both an
optionally substituted aryl. In some embodiments, at least one of
R.sup.6A and R.sup.7A can be an optionally substituted aryl. For
example, both R.sup.6A and R.sup.7A can be an optionally
substituted phenyl or an optionally substituted naphthyl. When
substituted, the substituted aryl can be substituted with 1, 2, 3
or more than 3 substituents. When more the two substituents are
present, the substituents can be the same or different. In some
embodiments, when at least one of R.sup.6A and R.sup.7A is a
substituted phenyl, the substituted phenyl can be a para-, ortho-
or meta-substituted phenyl.
[0099] In some embodiments, R.sup.6A and R.sup.7A can be both an
optionally substituted aryl(C.sub.1-6 alkyl). In some embodiments,
at least one of R.sup.6A and R.sup.7A can be an optionally
substituted aryl(C.sub.1-6 alkyl). For example, both R.sup.6A and
R.sup.7A can be an optionally substituted benzyl. When substituted,
the substituted benzyl group can be substituted with 1, 2, 3 or
more than 3 substituents. When more the two substituents are
present, the substituents can be the same or different. In some
embodiments, the aryl group of the aryl(C.sub.1-6 alkyl) can be a
para-, ortho- or meta-substituted phenyl.
[0100] In some embodiments, R.sup.6A and R.sup.7A can be both
##STR00036##
In some embodiments, at least one of R.sup.6A and R.sup.7A can
be
##STR00037##
In some embodiments, R.sup.25A can be hydrogen. In other
embodiments, R.sup.25A can be an optionally substituted C.sub.1-24
alkyl. In still other embodiments, R.sup.25A can be an optionally
substituted aryl. In some embodiments, R.sup.25A can be a C.sub.1-6
alkyl, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl,
isobutyl, tert-butyl, pentyl (branched and straight-chained), and
hexyl (branched and straight-chained).
[0101] In some embodiments, R.sup.6A and R.sup.7A can be both
##STR00038##
In some embodiments, at least one of R.sup.6A and R.sup.7A can
be
##STR00039##
In some embodiments, R.sup.29A can be hydrogen. In other
embodiments, R.sup.29A can be an optionally substituted C.sub.1-24
alkyl. In some embodiments, R.sup.29A can be a C.sub.1-4 alkyl,
such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl and
t-butyl. In still other embodiments, R.sup.29A can be an optionally
substituted aryl, such as an optionally substituted phenyl or an
optionally substituted naphthyl.
[0102] In some embodiments, R.sup.1A can be
##STR00040##
R.sup.6A can be
##STR00041##
[0103] R.sup.7A can be absent or hydrogen; R.sup.12A, R.sup.13A and
R.sup.14A can be independently absent or hydrogen; and m can be 0
or 1. In some embodiments, m can be 0, and R.sup.7A, R.sup.12A and
R.sup.13A can be independently absent or hydrogen. In other
embodiments, m can be 1, and R.sup.7A, R.sup.12A, R.sup.13A and
R.sup.14A can be independently absent or hydrogen. Those skilled in
the art understand that when m is 0, R.sup.6A can be diphosphate,
when Z.sup.1A is oxygen, or an alpha-thiodiphosphate, when Z.sup.1A
is sulfur. Likewise, those skilled in the art understand that when
m is 1, R.sup.6A can be triphosphate, when Z.sup.1A is oxygen, or
an alpha-thiotriphosphate, when Z.sup.1A is sulfur.
[0104] In some embodiments, R.sup.6A and R.sup.7A can be taken
together to form an optionally substituted
##STR00042##
For example, R.sup.1A can be an optionally substituted
##STR00043##
When substituted, the ring can be substituted 1, 2, 3 or 3 or more
times. When substituted with multiple substituents, the
substituents can be the same or different. In some embodiments,
when R.sup.1A is
##STR00044##
the ring can be substituted with an optionally substituted aryl
group and/or an optionally substituted heteroaryl. An example of a
suitable heteroaryl is pyridinyl. In some embodiments, R.sup.6A and
R.sup.7A can be taken together to form an optionally substitute
##STR00045##
such as
##STR00046##
wherein R.sup.32A can be an optionally substituted aryl, an
optionally substituted heteroaryl or an optionally substituted
heterocyclyl.
[0105] In some embodiments, R.sup.6A and R.sup.7A can be taken
together to form an optionally substituted
##STR00047##
wherein the oxygens connected to R.sup.6A and R.sup.7A, the
phosphorus and the moiety form a six-membered to ten-membered ring
system. Example of an optionally substituted
##STR00048##
include
##STR00049##
[0106] In some embodiments, R.sup.6A and R.sup.7A can be the same.
In some embodiments, R.sup.6A and R.sup.7A can be the
different.
[0107] In some embodiments, ZIA can be oxygen. In other
embodiments, ZIA can be sulfur.
[0108] In some embodiments, R.sup.1A can be
##STR00050##
In some embodiments, R.sup.8A can be selected from absent,
hydrogen, an optionally substituted C.sub.1-24 alkyl, an optionally
substituted C.sub.2-24 alkenyl, an optionally substituted
C.sub.2-24 alkynyl, an optionally substituted C.sub.3-6 cycloalkyl
and an optionally substituted C.sub.3-6 cycloalkenyl; and R.sup.9A
can be independently selected from an optionally substituted
C.sub.1-24 alkyl, an optionally substituted C.sub.2-24 alkenyl, an
optionally substituted C.sub.2-24 alkynyl, an optionally
substituted C.sub.3-6 cycloalkyl and an optionally substituted
C.sub.3-6 cycloalkenyl.
[0109] In some embodiments, R.sup.8A can be hydrogen, and R.sup.9A
can be an optionally substituted C.sub.1-6 alkyl. Examples of
suitable C.sub.1-6 alkyls include methyl, ethyl, n-propyl,
isopropyl, n-butyl, isobutyl, tert-butyl, pentyl (branched and
straight-chained), and hexyl (branched and straight-chained). In
other embodiments, R.sup.8A can be hydrogen, and R.sup.9A can be
NR.sup.30AR.sup.31A, wherein R.sup.30 and R.sup.31 can be
independently selected from hydrogen, an optionally substituted
C.sub.1-24 alkyl, an optionally substituted C.sub.2-24 alkenyl, an
optionally substituted C.sub.2-24 alkynyl, an optionally
substituted C.sub.3-6 cycloalkyl and an optionally substituted
C.sub.3-6 cycloalkenyl.
[0110] In some embodiments, R.sup.8A can be absent or hydrogen; and
R.sup.9A can be an optionally substituted N-linked amino acid or an
optionally substituted N-linked amino acid ester derivative. In
other embodiments, R.sup.8A can be an optionally substituted aryl;
and R.sup.9A can be an optionally substituted N-linked amino acid
or an optionally substituted N-linked amino acid ester derivative.
In still other embodiments, R.sup.8A can be an optionally
substituted heteroaryl; and R.sup.9A can be an optionally
substituted N-linked amino acid or an optionally substituted
N-linked amino acid ester derivative. In some embodiments, R.sup.9A
can be selected from alanine, asparagine, aspartate, cysteine,
glutamate, glutamine, glycine, proline, serine, tyrosine, arginine,
histidine, isoleucine, leucine, lysine, methionine, phenylalanine,
threonine, tryptophan, valine and ester derivatives thereof.
Examples of an optionally substituted N-linked amino acid ester
derivatives include optionally substituted versions of the
following: alanine isopropyl ester, alanine cyclohexyl ester,
alanine neopentyl ester, valine isopropyl ester and leucine
isopropyl ester. In some embodiments, R.sup.9A can have the
structure
##STR00051##
wherein R.sup.33A can be selected from hydrogen, an optionally
substituted C.sub.1-6-alkyl, an optionally substituted C.sub.3-6
cycloalkyl, an optionally substituted aryl, an optionally
substituted aryl(C.sub.1-6 alkyl) and an optionally substituted
haloalkyl; R.sup.34A can be selected from hydrogen, an optionally
substituted C.sub.1-6 alkyl, an optionally substituted C.sub.1-6
haloalkyl, an optionally substituted C.sub.3-6 cycloalkyl, an
optionally substituted C.sub.6 aryl, an optionally substituted
C.sub.10 aryl and an optionally substituted aryl(C.sub.1-6 alkyl);
and R.sup.35A can be hydrogen or an optionally substituted
C.sub.1-4-alkyl; or R.sup.34A and R.sup.35A can be taken together
to form an optionally substituted C.sub.3-6 cycloalkyl.
[0111] When R.sup.34A is substituted, R.sup.34A can be substituted
with one or more substituents selected from N-amido, mercapto,
alkylthio, an optionally substituted aryl, hydroxy, an optionally
substituted heteroaryl, O-carboxy, and amino. In some embodiments,
R.sup.34A can be an unsubstituted C.sub.1-6-alkyl, such as those
described herein. In some embodiments, R.sup.34A can be hydrogen.
In other embodiments, R.sup.34A can be methyl. In some embodiments,
R.sup.33A can be an optionally substituted C.sub.1-6 alkyl.
Examples of optionally substituted C.sub.1-6-alkyls include
optionally substituted variants of the following: methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl
(branched and straight-chained), and hexyl (branched and
straight-chained). In some embodiments, R.sup.33A can be methyl or
isopropyl. In some embodiments, R.sup.33A can be ethyl or
neopentyl. In other embodiments, R.sup.33A can be an optionally
substituted C.sub.3-6 cycloalkyl. Examples of optionally
substituted C.sub.3-6 cycloalkyl include optionally substituted
variants of the following: cyclopropyl, cyclobutyl, cyclopentyl,
and cyclohexyl. In an embodiment, R.sup.33A can be an optionally
substituted cyclohexyl. In still other embodiments, R.sup.33A can
be an optionally substituted aryl, such as phenyl and naphthyl. In
yet still other embodiments, R.sup.33A can be an optionally
substituted aryl(C.sub.1-6 alkyl). In some embodiments, R.sup.33A
can be an optionally substituted benzyl. In some embodiments,
R.sup.33A can be an optionally substituted C.sub.1-6 haloalkyl, for
example, CF.sub.3. In some embodiments, R.sup.35A can be hydrogen.
In other embodiments, R.sup.35A can be an optionally substituted
C.sub.1-4-alkyl, such as methyl, ethyl, n-propyl, isopropyl,
n-butyl, isobutyl and tert-butyl. In an embodiment, R.sup.35A can
be methyl. In some embodiments, R.sup.34A and R.sup.35A can be
taken together to form an optionally substituted C.sub.3-6
cycloalkyl. Examples of optionally substituted C.sub.3-6 cycloalkyl
include optionally substituted variants of the following:
cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Depending on
the groups that are selected for R.sup.34A and R.sup.35A, the
carbon to which R.sup.34A and R.sup.35A are attached may be a
chiral center. In some embodiment, the carbon to which R.sup.34A
and R.sup.35A are attached may be a (R)-chiral center. In other
embodiments, the carbon to which R.sup.34A and R.sup.35A are
attached may be a (S)-chiral center.
[0112] In some embodiments, when R.sup.1A is
##STR00052##
Z.sup.2A can be O (oxygen). In other embodiments, when R.sup.1A
is
##STR00053##
Z.sup.2A can be S (sulfur).
[0113] In some embodiments, R.sup.1A can be
##STR00054##
In some embodiments, R.sup.10A and R.sup.11A can be both an
optionally substituted N-linked amino acid or an optionally
substituted N-linked amino acid ester derivative. In some
embodiments, R.sup.10A and R.sup.11A can be independently selected
from alanine, asparagine, aspartate, cysteine, glutamate,
glutamine, glycine, proline, serine, tyrosine, arginine, histidine,
isoleucine, leucine, lysine, methionine, phenylalanine, threonine,
tryptophan, valine and ester derivatives thereof. In some
embodiments, R.sup.10A and R.sup.11A can be an optionally
substituted version of the following: alanine isopropyl ester,
alanine cyclohexyl ester, alanine neopentyl ester, valine isopropyl
ester and leucine isopropyl ester. In some embodiments, R.sup.10A
and R.sup.11A can independently have the structure
##STR00055##
wherein R.sup.36A can be selected from hydrogen, an optionally
substituted C.sub.1-6-alkyl, an optionally substituted C.sub.3-6
cycloalkyl, an optionally substituted aryl, an optionally
substituted aryl(C.sub.1-6 alkyl) and an optionally substituted
haloalkyl; R.sup.37A can be selected from hydrogen, an optionally
substituted C.sub.1-6 alkyl, an optionally substituted C.sub.1-6
haloalkyl, an optionally substituted C.sub.3-6 cycloalkyl, an
optionally substituted C.sub.6 aryl, an optionally substituted
C.sub.10 aryl and an optionally substituted aryl(C.sub.1-6 alkyl);
and R.sup.38A can be hydrogen or an optionally substituted
C.sub.1-4-alkyl; or R.sup.37A and R.sup.38A can be taken together
to form an optionally substituted C.sub.3-6 cycloalkyl.
[0114] When R.sup.37A is substituted, R.sup.37A can be substituted
with one or more substituents selected from N-amido, mercapto,
alkylthio, an optionally substituted aryl, hydroxy, an optionally
substituted heteroaryl, O-carboxy, and amino. In some embodiments,
R.sup.37A can be an unsubstituted C.sub.1-6-alkyl, such as those
described herein. In some embodiments, R.sup.37A can be hydrogen.
In other embodiments, R.sup.37A can be methyl. In some embodiments,
R.sup.36A can be an optionally substituted C.sub.1-6 alkyl.
Examples of optionally substituted C.sub.1-6-alkyls include
optionally substituted variants of the following: methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl
(branched and straight-chained), and hexyl (branched and
straight-chained). In some embodiments, R.sup.36A can be methyl or
isopropyl. In some embodiments, R.sup.36A can be ethyl or
neopentyl. In other embodiments, R.sup.36A can be an optionally
substituted C.sub.3-6 cycloalkyl. Examples of optionally
substituted C.sub.3-6 cycloalkyl include optionally substituted
variants of the following: cyclopropyl, cyclobutyl, cyclopentyl,
and cyclohexyl. In an embodiment, R.sup.36A can be an optionally
substituted cyclohexyl. In still other embodiments, R.sup.36A can
be an optionally substituted aryl, such as phenyl and naphthyl. In
yet still other embodiments, R.sup.36A can be an optionally
substituted aryl(C.sub.1-6 alkyl). In some embodiments, R.sup.36A
can be an optionally substituted benzyl. In some embodiments,
R.sup.36A can be an optionally substituted C.sub.1-6 haloalkyl, for
example, CF.sub.3. In some embodiments, R.sup.38A can be hydrogen.
In other embodiments, R.sup.38A can be an optionally substituted
C.sub.1-4-alkyl, such as methyl, ethyl, n-propyl, isopropyl,
n-butyl, isobutyl and tert-butyl. In an embodiment, R.sup.38A can
be methyl. In some embodiments, R.sup.37A and R.sup.38A can be
taken together to form an optionally substituted C.sub.3-6
cycloalkyl. Examples of optionally substituted C.sub.3-6 cycloalkyl
include optionally substituted variants of the following:
cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Depending on
the groups that are selected for R.sup.37A and R.sup.38A, the
carbon to which R.sup.37A and R.sup.38A are attached may be a
chiral center. In some embodiment, the carbon to which R.sup.37A
and R.sup.38A are attached may be a (R)-chiral center. In other
embodiments, the carbon to which R.sup.37A and R.sup.38A are
attached may be a (S)-chiral center.
[0115] Examples of suitable
##STR00056##
groups include the following:
##STR00057##
[0116] In some embodiments, R.sup.10A and R.sup.11A can be the
same. In some embodiments, R.sup.10A and R.sup.11A can be the
different.
[0117] In some embodiments, Z.sup.3A can be O (oxygen). In other
embodiments, Z.sup.3A can be S (sulfur).
[0118] In some embodiments, R.sup.1A can be hydrogen. In some
embodiments, R.sup.1A can be an optionally substituted acyl. In
other embodiments, R.sup.1A can be --C(.dbd.O)R.sup.39A wherein
R.sup.39A can be selected from an optionally substituted C.sub.1-12
alkyl, an optionally substituted C.sub.2-12 alkenyl, an optionally
substituted C.sub.2-12 alkynyl, an optionally substituted C.sub.3-8
cycloalkyl, an optionally substituted C.sub.5-8 cycloalkenyl, an
optionally substituted C.sub.6-10 aryl, an optionally substituted
heteroaryl, an optionally substituted heterocyclyl, an optionally
substituted aryl(C.sub.1-6 alkyl), an optionally substituted
heteroaryl(C.sub.1-6 alkyl) and an optionally substituted
heterocyclyl(C.sub.1-6 alkyl). In some embodiments, R.sup.39A can
be a substituted C.sub.1-12 alkyl. In other embodiments, R.sup.39A
can be an unsubstituted C.sub.1-12 alkyl.
[0119] In still other embodiments, R.sup.1A can be an optionally
substituted O-linked amino acid. Examples of suitable O-linked
amino acids include alanine, asparagine, aspartate, cysteine,
glutamate, glutamine, glycine, proline, serine, tyrosine, arginine,
histidine, isoleucine, leucine, lysine, methionine, phenylalanine,
threonine, tryptophan and valine. Additional examples of suitable
amino acids include, but are not limited to, ornithine, hypusine,
2-aminoisobutyric acid, dehydroalanine, gamma-aminobutyric acid,
citrulline, beta-alanine, alpha-ethyl-glycine, alpha-propyl-glycine
and norleucine. In some embodiments, the O-linked amino acid can
have the structure
##STR00058##
wherein R.sup.40A can be selected from hydrogen, an optionally
substituted C.sub.1-6 alkyl, an optionally substituted C.sub.1-6
haloalkyl, an optionally substituted C.sub.3-6 cycloalkyl, an
optionally substituted C.sub.6 aryl, an optionally substituted
C.sub.10 aryl and an optionally substituted aryl(C.sub.1-6 alkyl);
and R.sup.41A can be hydrogen or an optionally substituted
C.sub.1-4-alkyl; or R.sup.40A and R.sup.41A can be taken together
to form an optionally substituted C.sub.3-6 cycloalkyl. Those
skilled in the art understand that when R.sup.1A is an optionally
substituted O-linked amino acid, the oxygen of R.sup.1A O-- of
Formula (I) is part of the optionally substituted O-linked amino
acid. For example, when R.sup.1A is
##STR00059##
the oxygen indicated with "*" is the oxygen of R.sup.1AO-- of
Formula (I).
[0120] When R.sup.40A is substituted, R.sup.40A can be substituted
with one or more substituents selected from N-amido, mercapto,
alkylthio, an optionally substituted aryl, hydroxy, an optionally
substituted heteroaryl, O-carboxy, and amino. In some embodiments,
R.sup.40A can be an unsubstituted C.sub.1-6-alkyl, such as those
described herein. In some embodiments, R.sup.40A can be hydrogen.
In other embodiments, R.sup.40A can be methyl. In some embodiments,
R.sup.41A can be hydrogen. In other embodiments, R.sup.41A can be
an optionally substituted C.sub.1-4-alkyl, such as methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl. In an
embodiment, R.sup.41A can be methyl. Depending on the groups that
are selected for R.sup.40A and R.sup.41A, the carbon to which
R.sup.40A and R.sup.41A are attached may be a chiral center. In
some embodiment, the carbon to which R.sup.40A and R.sup.41A are
attached may be a (R)-chiral center. In other embodiments, the
carbon to which R.sup.40A and R.sup.41A are attached may be a
(S)-chiral center.
[0121] Examples of suitable
##STR00060##
include the following:
##STR00061##
[0122] In some embodiments, the dashed line () can be a single
bond, R.sup.2A can be CH.sub.2, and R.sup.3A can be O (oxygen).
When the dashed line () is a single bond, R.sup.2A is CH.sub.2, and
R.sup.3A is O (oxygen), a 4-membered ring is formed that includes
the 4'-carbon and 3'-carbon of the pentose ring. In other
embodiments, the dashed line () can be absent, R.sup.2A can be
selected from an optionally substituted C.sub.1-6 alkyl, an
optionally substituted C.sub.2-6 alkenyl, an optionally substituted
C.sub.2-6 alkynyl, an optionally substituted --O--C.sub.1-6 alkyl,
an optionally substituted --O--C.sub.3-6 alkenyl, an optionally
substituted --O--C.sub.3-6 alkynyl and cyano, and R.sup.3A can be
selected from OH, --OC(.dbd.O)R.sup.''A and an optionally
substituted O-linked amino acid.
[0123] Various groups can be attached to the 4'-position of the
pentose ring. In some embodiments, R.sup.2A can be an optionally
substituted C.sub.1-6 alkyl. Examples of suitable C.sub.1-6 alkyls
include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,
tert-butyl, pentyl (branched and straight-chained), and hexyl
(branched and straight-chained). In some embodiments, R.sup.2A can
be an unsubstituted C.sub.1-6 alkyl. In other embodiments, R.sup.2A
can be a substituted C.sub.1-6 alkyl. For example, R.sup.2A can be
a halogen substituted C.sub.1-6 alkyl, a hydroxy substituted
C.sub.1-6 alkyl, an alkoxy substituted C.sub.1-6 alkyl or a
sulfenyl substituted C.sub.1-6 alkyl (for example, --C.sub.1-6
alkyl-S--C.sub.1-6 alkyl). In other embodiments, R.sup.2A can be a
C.sub.1-6 haloalkyl. In other embodiments, R.sup.2A can be an
optionally substituted C.sub.2-6 alkenyl. In some embodiments,
R.sup.2A can be a substituted C.sub.2-6 alkenyl. In other
embodiments, R.sup.2A can be an unsubstituted C.sub.2-6 alkenyl.
For example, R.sup.2A can be ethenyl, propenyl or allenyl. In still
other embodiments, R.sup.2A can be an optionally substituted
C.sub.2-6 alknyl. In some embodiments, R.sup.2A can be a
substituted C.sub.2-6 alkynyl. In other embodiments, R.sup.2A can
be an unsubstituted C.sub.2-6 alkynyl. Suitable C.sub.2-6 alkynyls
include ethynyl and propynyl. In yet still other embodiments,
R.sup.2A can be an optionally substituted C.sub.3-6 cycloalkyl. In
some embodiments, R.sup.2A can be a substituted C.sub.3-6
cycloalkyl. In other embodiments, R.sup.2A can be an unsubstituted
C.sub.3-6 cycloalkyl. A non-limiting list of C.sub.3-6 cycloalkyls
include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. In
some embodiments, R.sup.2A can be an optionally substituted
--O--C.sub.1-6 alkyl. In some embodiments, R.sup.2A can be a
substituted --O--C.sub.1-6 alkyl. In other embodiments, R.sup.2A
can be an unsubstituted --O--C.sub.1-6 alkyl. Examples of suitable
O--C.sub.1-6 alkyl groups include methoxy, ethoxy, n-propoxy,
iso-propoxy, n-butoxy, isobutoxy, tert-butoxy, pentoxy (branched
and straight-chained), and hexoxy (branched and straight-chained).
In other embodiments, R.sup.2A can be an optionally substituted
--O--C.sub.3-6 alkenyl. In some embodiments, R.sup.2A can be a
substituted --O--C.sub.3-6 alkenyl. In other embodiments, R.sup.2A
can be an unsubstituted --O--C.sub.3-6 alkenyl. In still other
embodiments, R.sup.2A can be an optionally substituted
--O--C.sub.3-6 alkynyl. In some embodiments, R.sup.2A can be a
substituted --O--C.sub.3-6 alkynyl. In other embodiments, R.sup.2A
can be an unsubstituted --O--C.sub.3-6 alkynyl. In yet still other
embodiments, R.sup.2A can be cyano.
[0124] The groups attached to the 3'-position of the pentose ring
can vary. In some embodiments, including those of paragraph [0120],
R.sup.3A can be OH. In other embodiments, including those of
paragraph [0120], R.sup.3A can be an optionally substituted
0-linked amino acid. Examples of suitable O-linked amino acids
include alanine, asparagine, aspartate, cysteine, glutamate,
glutamine, glycine, proline, serine, tyrosine, arginine, histidine,
isoleucine, leucine, lysine, methionine, phenylalanine, threonine,
tryptophan and valine. Additional examples of suitable amino acids
include, but are not limited to, ornithine, hypusine,
2-aminoisobutyric acid, dehydroalanine, gamma-aminobutyric acid,
citrulline, beta-alanine, alpha-ethyl-glycine, alpha-propyl-glycine
and norleucine. In some embodiments, the O-linked amino acid can
have the structure
##STR00062##
wherein R.sup.42A can be selected from hydrogen, an optionally
substituted C.sub.1-6 alkyl, an optionally substituted C.sub.1-6
haloalkyl, an optionally substituted C.sub.3-6 cycloalkyl, an
optionally substituted C.sub.6 aryl, an optionally substituted
C.sub.10 aryl and an optionally substituted aryl(C.sub.1-6 alkyl);
and R.sup.43A can be hydrogen or an optionally substituted
C.sub.1-4-alkyl; or R.sup.4 and R.sup.43A can be taken together to
form an optionally substituted C.sub.3-6 cycloalkyl.
[0125] When R.sup.42A is substituted, R.sup.42A can be substituted
with one or more substituents selected from N-amido, mercapto,
alkylthio, an optionally substituted aryl, hydroxy, an optionally
substituted heteroaryl, O-carboxy, and amino. In some embodiments,
R.sup.42A can be an unsubstituted C.sub.1-6-alkyl, such as those
described herein. In some embodiments, R.sup.42A can be hydrogen.
In other embodiments, R.sup.42A can be methyl. In some embodiments,
R.sup.43A can be hydrogen. In other embodiments, R.sup.43A Can be
an optionally substituted C.sub.1-4-alkyl, such as methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl. In an
embodiment, R.sup.43A can be methyl. Depending on the groups that
are selected for R.sup.42A and R.sup.43A, the carbon to which
R.sup.42A and R.sup.43A are attached may be a chiral center. In
some embodiment, the carbon to which R.sup.42A and R.sup.43A are
attached may be a (R)-chiral center. In other embodiments, the
carbon to which R.sup.42A and R.sup.43A are attached may be a
(S)-chiral center.
[0126] Examples of suitable
##STR00063##
include the following:
##STR00064##
[0127] In still other embodiments, including those of paragraph
[0120], R.sup.3A Can be --OC(.dbd.O)R.sup.''A, wherein R.sup.''A
can be an optionally substituted C.sub.1-24 alkyl. In some
embodiments, R.sup.''A can be a substituted C.sub.1-8 alkyl. In
other embodiments, R.sup.''A can be an unsubstituted C.sub.1-8
alkyl. In still other embodiments, including those of paragraph
[0120], R.sup.3A can be an optionally substituted --O-acyl. In yet
still other embodiments, including those of paragraph [0120],
R.sup.3A can be --C(.dbd.O)R.sup.4A, wherein R.sup.44A can be
selected from an optionally substituted C.sub.1-12 alkyl, an
optionally substituted C.sub.2-12 alkenyl, an optionally
substituted C.sub.2-12 alkynyl, an optionally substituted C.sub.3-8
cycloalkyl, an optionally substituted C.sub.5-8 cycloalkenyl, an
optionally substituted C.sub.6-10 aryl, an optionally substituted
heteroaryl, an optionally substituted heterocyclyl, an optionally
substituted aryl(C.sub.1-6 alkyl), an optionally substituted
heteroaryl(C.sub.1-6 alkyl) and an optionally substituted
heterocyclyl(C.sub.1-6 alkyl). In some embodiments, R.sup.44A can
be a substituted C.sub.1-12 alkyl. In other embodiments, R.sup.44A
can be an unsubstituted C.sub.1-12 alkyl.
[0128] Various substituents can be present at the 2'-position of
the pentose ring. In some embodiments, R.sup.5A can be hydrogen. In
other embodiments, R.sup.5A can be halogen, for example, fluoro. In
some embodiments, R.sup.4A can be halogen, such as fluoro. In some
embodiments, R.sup.5A can be hydrogen and R.sup.4A can be halogen.
In other embodiments, R.sup.4A and R.sup.5A can both be
halogen.
[0129] In some embodiments, can be a single bond, R.sup.4A can be
fluoro, R.sup.5A can be hydrogen and R.sup.2A can be a C.sub.1-6
haloalkyl. In some embodiments, can be a single bond, R.sup.4A can
be fluoro, R.sup.5A can be hydrogen, R.sup.2A can be a C.sub.1-6
haloalkyl and B.sup.1A can be cytosine.
[0130] In some embodiments, R.sup.2A cannot be methoxy. In some
embodiments, R.sup.2A cannot be methoxy when B.sup.1A is
substituted or unsubstituted uracil. In some embodiments, B.sup.1A
is substituted or unsubstituted cytosine. In other embodiments,
B.sup.1A is substituted or unsubstituted thymine. In still other
embodiments, B.sup.1A cannot be an unsubstituted uracil. In some
embodiments, R.sup.2A cannot be methoxy when Z.sup.1A is
##STR00065##
wherein R.sup.8A is an unsubstituted C.sub.1-6 alkyl or a
para-substituted phenyl; and R.sup.9A is an optionally substituted
N-linked amino acid or an optionally substituted N-linked amino
acid ester derivative. In some embodiments, R.sup.2A cannot be
methoxy when Z.sup.1A is
##STR00066##
In some embodiments, R.sup.2A cannot be an alkoxy (for example,
when Z.sup.1A is
##STR00067##
In some embodiments, B.sup.1A cannot be cytosine when R.sup.2A is
an unsubstituted alkenyl or an unsubstituted alkynyl. In some
embodiments, B.sup.1A cannot be thymine when R.sup.2A is an
optionally substituted alkyl. In some embodiments, R.sup.2A cannot
be an unsubstituted alkoxy (such as methoxy), an optionally
substituted alkenyl (such as allenyl), an unsubstituted alkynyl
(such as ethynyl) or a C alkyl substituted with a non-halogen
substituent. In some embodiments, R.sup.2A cannot be an
unsubstituted alkoxy (such as methoxy), an optionally substituted
alkenyl (such as allenyl), an optionally substituted substituted
alkynyl (such as ethynyl) or a C.sub.1-4 alkyl substituted with a
non-halogen substituent. In some embodiments R.sup.1A cannot be H.
In some embodiments R.sup.1A cannot be H when B.sup.1A is an
optionally substituted cytosine or an optionally substituted
thymine.
[0131] Various optionally substituted heterocyclic bases can be
attached to the pentose ring. In some embodiments, one or more of
the amine and/or amino groups may be protected with a suitable
protecting group. For example, an amino group may be protected by
transforming the amine and/or amino group to an amide or a
carbamate. In some embodiments, an optionally substituted
heterocyclic base or an optionally substituted heterocyclic base
with one or more protected amino groups can have one of the
following structures:
##STR00068##
wherein: R.sup.A2 can be selected from hydrogen, halogen and
NHR.sup.J2, wherein R.sup.J2 can be selected from hydrogen,
--C(.dbd.O)R.sup.K2 and --C(.dbd.O)OR.sup.L2; R.sup.B2 can be
halogen or NHR.sup.W2, wherein R.sup.W2 can be selected from
hydrogen, an optionally substituted C.sub.1-6 alkyl, an optionally
substituted C.sub.2-6 alkenyl, an optionally substituted C.sub.3-8
cycloalkyl, --C(.dbd.O)R.sup.M2 and --C(.dbd.O)OR.sup.N2; R.sup.C2
can be hydrogen or NHR.sup.O2, wherein R.sup.O2 can be selected
from hydrogen, --C(.dbd.O)R.sup.P2 and --C(.dbd.O)OR.sup.Q2;
R.sup.D2 can be selected from hydrogen, halogen, an optionally
substituted C.sub.1-6 alkyl, an optionally substituted C.sub.2-6
alkenyl and an optionally substituted C.sub.2-6 alkynyl; R.sup.E2
can be selected from hydrogen, hydroxy, an optionally substituted
C.sub.1-6 alkyl, an optionally substituted C.sub.3-8 cycloalkyl,
--C(.dbd.O)R.sup.R2 and --C(.dbd.O)OR.sup.S2; R.sup.F2 can be
selected from hydrogen, halogen, an optionally substituted
C.sub.1-6 alkyl, an optionally substituted C.sub.2-6 alkenyl and an
optionally substituted C.sub.2-6 alkynyl; Y.sup.2 and Y.sup.3 can
be independently N (nitrogen) or CR.sup.I2, wherein R.sup.I2 can be
selected from hydrogen, halogen, an optionally substituted
C.sub.1-6-alkyl, an optionally substituted C.sub.2-6-alkenyl and an
optionally substituted C.sub.2-6-alkynyl; R.sup.G2 can be an
optionally substituted C.sub.1-6 alkyl; RH.sup.2 can be hydrogen or
NHR.sup.T2, wherein R.sup.T2 can be independently selected from
hydrogen, --C(.dbd.O)R.sup.U2 and --C(.dbd.O)OR.sup.V2; and
R.sup.K2, R.sup.L2, R.sup.M2, R.sup.N2, R.sup.P2, R.sup.Q2,
R.sup.R2, R.sup.S2, R.sup.U2 and R.sup.V2 can be independently
selected from C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl, C.sub.3-6 cycloalkyl, C.sub.3-6 cycloalkenyl, C.sub.6-10
aryl, heteroaryl, heteroalicyclyl, aryl(C.sub.1-6 alkyl),
heteroaryl(C.sub.1-6 alkyl) and heteroalicyclyl(C.sub.1-6 alkyl).
In some embodiments, the structures shown above can be modified by
replacing one or more hydrogens with substituents selected from the
list of substituents provided for the definition of
"substituted."
[0132] In some embodiments, B.sup.1A can be
##STR00069##
In other embodiments, B.sup.1A can be
##STR00070##
In still other embodiments, B.sup.1A can be
##STR00071##
such as
##STR00072##
In yet still other embodiments, B.sup.1A can be
##STR00073##
for example,
##STR00074##
In some embodiments, R.sup.D2 can be hydrogen. In other
embodiments, B.sup.1A can be
##STR00075##
In some embodiments, R.sup.B2 can be NH.sub.2. In other
embodiments, R.sup.B2 can be NHR.sup.W2, wherein R.sup.W2 can be
--C(.dbd.O)R.sup.M2 or --C(.dbd.O)OR.sup.N2. In still other
embodiments, B.sup.1A can be
##STR00076##
In some embodiments, B.sup.1A can be
##STR00077##
[0133] In some embodiments, a compound of Formula (I) can have a
structure selected from one of the following:
##STR00078## ##STR00079## ##STR00080##
or a pharmaceutically acceptable salt of the foregoing. In some
embodiments of this paragraph, B.sup.1A can be an optionally
substituted purine base. In other embodiments of this paragraph,
B.sup.1A can be an optionally substituted pyrimidine base. In some
embodiments of this paragraph, B.sup.1A can be guanine. In other
embodiments of this paragraph, B.sup.1A can be thymine. In still
other embodiments of this paragraph, B.sup.1A can be cytosine. In
yet still other embodiments of this paragraph, B.sup.1A can be
uracil. In some embodiments of this paragraph, B.sup.1A can be
adenine. In some embodiments of this paragraph, R.sup.1A can be
hydrogen. In other embodiments of this paragraph, R.sup.1A can be
an optionally substituted acyl. In still other embodiments of this
paragraph, R.sup.1A can be mono-, di- or tri-phosphate. In yet
other embodiments of this paragraph, R.sup.1A can be
phosphoroamidate. In some embodiments of this paragraph, R.sup.1A
can be an acyloxyalkyl ester phosphate prodrug.
[0134] In some embodiments, the compound can be a compound of
Formula (II), or a pharmaceutically acceptable salt thereof,
wherein: B.sup.1B can be an optionally substituted heterocyclic
base or an optionally substituted heterocyclic base with a
protected amino group; R.sup.1B can be selected from O.sup.-,
OH,
##STR00081##
an optionally substituted N-linked amino acid and an optionally
substituted N-linked amino acid ester derivative; R.sup.2B can be
selected from an optionally substituted C.sub.1-6 alkyl, an
optionally substituted C.sub.2-6 alkenyl, an optionally substituted
C.sub.2-6 alkynyl, an optionally substituted --O--C.sub.1-6 alkyl,
an optionally substituted --O--C.sub.3-6 alkenyl, an optionally
substituted --O--C.sub.3-6 alkynyl and cyano; R.sup.3B can be a
halogen; R.sup.4B can be hydrogen or halogen; R.sup.5B, R.sup.6B,
R.sup.8B and R.sup.9B can be independently selected from hydrogen,
an optionally substituted C.sub.1-24 alkyl and an optionally
substituted aryl; R.sup.7B and R.sup.10B can be independently
selected from hydrogen, an optionally substituted C.sub.1-24 alkyl,
an optionally substituted aryl, an optionally substituted
--O--C.sub.1-24 alkyl and an optionally substituted --O-aryl;
R.sup.11B can be selected from hydrogen, an optionally substituted
C.sub.1-24 alkyl and an optionally substituted aryl; Z.sup.1B and
Z.sup.2B can be independently O or S.
[0135] In some embodiments, R.sup.1B can be O.sup.-. In other
embodiments, R.sup.1B can be OH.
[0136] In some embodiments, R.sup.1B can be
##STR00082##
wherein R.sup.5B and R.sup.6B can be independently selected from
hydrogen, an optionally substituted C.sub.1-24 alkyl and an
optionally substituted aryl; and R.sup.7B can be selected from
hydrogen, an optionally substituted C.sub.1-24 alkyl, an optionally
substituted aryl, an optionally substituted --O--C.sub.1-24 alkyl
and an optionally substituted --O-aryl. In some embodiments,
R.sup.5B and R.sup.6B can be hydrogen. In other embodiments, at
least one of R.sup.5B and R.sup.6B can be an optionally substituted
C.sub.1-24 alkyl or an optionally substituted aryl. In some
embodiments, R.sup.7B can be an optionally substituted C.sub.1-24
alkyl. In other embodiments, R.sup.7B can be an optionally
substituted aryl. In still other embodiments, R.sup.7B can be an
optionally substituted --O--C.sub.1-24 alkyl or an optionally
substituted --O-aryl.
[0137] In some embodiments, R.sup.1B can be
##STR00083##
wherein R.sup.8B and R.sup.9B can be independently selected from
hydrogen, an optionally substituted C.sub.1-24 alkyl and an
optionally substituted aryl; R.sup.10B can be independently
selected from hydrogen, an optionally substituted C.sub.1-24 alkyl,
an optionally substituted aryl, an optionally substituted
--O--C.sub.1-24 alkyl and an optionally substituted --O-aryl; and
Z.sup.2B can be independently O (oxygen) or S (sulfur). In some
embodiments, R.sup.8B and R.sup.9B can be hydrogen. In other
embodiments, at least one of R.sup.8B and R.sup.9B can be an
optionally substituted C.sub.1-24 alkyl or an optionally
substituted aryl. In some embodiments, R.sup.10B can be an
optionally substituted C.sub.1-24 alkyl. In other embodiments,
R.sup.10B can be an optionally substituted aryl. In still other
embodiments, R.sup.10B can be an optionally substituted
--O--C.sub.1-24 alkyl or an optionally substituted --O-aryl. In
some embodiments, Z.sup.2B can be O (oxygen). In other embodiments,
Z.sup.2B can be or S (sulfur). In some embodiments, R.sup.1B can be
isopropylcarbonyloxymethyloxy. In some embodiments, R.sup.1B can be
pivaloyloxymethyloxy.
[0138] In some embodiments, R.sup.1B can be
##STR00084##
In some embodiments, R.sup.11B can be hydrogen. In other
embodiments, R.sup.11B can be an optionally substituted C.sub.1-24
alkyl. In still other embodiments, R.sup.11B can be an optionally
substituted aryl. In some embodiments, R.sup.11B can be a C.sub.1-6
alkyl, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl,
isobutyl, tert-butyl, pentyl (branched and straight-chained), and
hexyl (branched and straight-chained).
[0139] In some embodiments, R.sup.1B can be an optionally
substituted N-linked amino acid or an optionally substituted
N-linked amino acid ester derivative. For example, R.sup.1B can be
optionally substituted version of the following: alanine,
asparagine, aspartate, cysteine, glutamate, glutamine, glycine,
proline, serine, tyrosine, arginine, histidine, isoleucine,
leucine, lysine, methionine, phenylalanine, threonine, tryptophan,
valine and ester derivatives thereof. In some embodiments, R.sup.1B
can be selected from alanine isopropyl ester, alanine cyclohexyl
ester, alanine neopentyl ester, valine isopropyl ester and leucine
isopropyl ester. In some embodiments, R.sup.1B can have the
structure
##STR00085##
wherein R.sup.12B can be selected from hydrogen, an optionally
substituted C.sub.1-6-alkyl, an optionally substituted C.sub.3-6
cycloalkyl, an optionally substituted aryl, an optionally
substituted aryl(C.sub.1-6 alkyl) and an optionally substituted
haloalkyl; R.sup.13B can be selected from hydrogen, an optionally
substituted C.sub.1-6 alkyl, an optionally substituted C.sub.1-6
haloalkyl, an optionally substituted C.sub.3-6 cycloalkyl, an
optionally substituted C.sub.6 aryl, an optionally substituted
C.sub.10 aryl and an optionally substituted aryl(C.sub.1-6 alkyl);
and R.sup.14B can be hydrogen or an optionally substituted
C.sub.1-4-alkyl; or R.sup.13B and R.sup.14B can be taken together
to form an optionally substituted C.sub.3-6 cycloalkyl.
[0140] When R.sup.13B is substituted, R.sup.13B can be substituted
with one or more substituents selected from N-amido, mercapto,
alkylthio, an optionally substituted aryl, hydroxy, an optionally
substituted heteroaryl, O-carboxy, and amino. In some embodiments,
R.sup.13B can be an unsubstituted C.sub.1-6-alkyl, such as those
described herein. In some embodiments, R.sup.13B can be hydrogen.
In other embodiments, R.sup.13B can be methyl. In some embodiments,
R.sup.12B can be an optionally substituted C.sub.1-6 alkyl.
Examples of optionally substituted C.sub.1-6-alkyls include
optionally substituted variants of the following: methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl
(branched and straight-chained), and hexyl (branched and
straight-chained). In some embodiments, R.sup.12B can be methyl or
isopropyl. In some embodiments, R.sup.12B can be ethyl or
neopentyl. In other embodiments, R.sup.12B can be an optionally
substituted C.sub.3-6 cycloalkyl. Examples of optionally
substituted C.sub.3 cycloalkyl include optionally substituted
variants of the following: cyclopropyl, cyclobutyl, cyclopentyl,
and cyclohexyl. In an embodiment, R.sup.12B can be an optionally
substituted cyclohexyl. In still other embodiments, R.sup.12B can
be an optionally substituted aryl, such as phenyl and naphthyl. In
yet still other embodiments, R.sup.12B co be an optionally
substituted aryl(C.sub.1-6 alkyl). In some embodiments, R.sup.12B
can be an optionally substituted benzyl. In some embodiments,
R.sup.12B can be an optionally substituted C.sub.1_haloalkyl, for
example, CF.sub.3. In some embodiments, R.sup.14B can be hydrogen.
In other embodiments, R.sup.14B can be an optionally substituted
C.sub.1-4-alkyl, such as methyl, ethyl, n-propyl, isopropyl,
n-butyl, isobutyl and tert-butyl. In an embodiment, R.sup.14B can
be methyl. In some embodiments, R.sup.13B and R.sup.14B can be
taken together to form an optionally substituted C.sub.3-6
cycloalkyl. Examples of optionally substituted C.sub.3-6 cycloalkyl
include optionally substituted variants of the following:
cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Depending on
the groups that are selected for R.sup.13B and R.sup.14B, the
carbon to which R.sup.13B and R.sup.14B are attached may be a
chiral center. In some embodiment, the carbon to which R.sup.13B
and R.sup.14B are attached may be a (R)-chiral center. In other
embodiments, the carbon to which R.sup.13B and R.sup.14B are
attached may be a (S)-chiral center.
[0141] Examples of suitable
##STR00086##
groups include the following:
##STR00087##
[0142] A variety of substituents can be present at the 4'-position
of the pentose ring. In some embodiments, R.sup.2B can be an
optionally substituted C.sub.1-6 alkyl. Examples of suitable
C.sub.1-6 alkyls include methyl, ethyl, n-propyl, isopropyl,
n-butyl, isobutyl, tert-butyl, pentyl (branched and
straight-chained), and hexyl (branched and straight-chained). In
some embodiments, R.sup.2B can be an unsubstituted C.sub.1-6 alkyl.
In other embodiments, R.sup.2B can be a substituted C.sub.1-6
alkyl. For example, R.sup.2B can be a halogen substituted C.sub.1-6
alkyl, a hydroxy substituted C.sub.1-6 alkyl, an alkoxy substituted
C.sub.1-6 alkyl or a sulfenyl substituted C.sub.1-6 alkyl (for
example, --C.sub.1-6 alkyl-S--C.sub.1-6 alkyl). In other
embodiments, R.sup.2B can be a C.sub.1-6 haloalkyl. In other
embodiments, R.sup.2B can be an optionally substituted C.sub.2-6
alkenyl. In some embodiments, R.sup.2B can be a substituted
C.sub.2-6 alkenyl. In other embodiments, R.sup.2B can be an
unsubstituted C.sub.2-6 alkenyl. For example, R.sup.2B can be
ethenyl, propenyl or allenyl. In still other embodiments, R.sup.2B
can be an optionally substituted C.sub.2-6 alkynyl. In some
embodiments, R.sup.2B can be a substituted C.sub.2-6 alkynyl. In
other embodiments, R.sup.2B can be an unsubstituted C.sub.2-6
alknyl. Suitable C.sub.2-6 alkynyls include ethynyl and propynyl.
In yet still other embodiments, R.sup.2B can be an optionally
substituted C.sub.3-6 cycloalkyl. In some embodiments, R.sup.2B can
be a substituted C.sub.3-6 cycloalkyl. In other embodiments,
R.sup.2B can be an unsubstituted C.sub.3-6 cycloalkyl. A
non-limiting list of C.sub.3-6 cycloalkyls include cyclopropyl,
cyclobutyl, cyclopentyl and cyclohexyl. In some embodiments,
R.sup.2B can be an optionally substituted --O--C.sub.1-6 alkyl. In
some embodiments, R.sup.2B can be a substituted --O--C.sub.1-6
alkyl. In other embodiments, R.sup.2B can be an unsubstituted
--O--C.sub.1-6 alkyl. Examples of suitable O--C.sub.1-6 alkyl
groups include methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy,
isobutoxy, tert-butoxy, pentoxy (branched and straight-chained),
and hexoxy (branched and straight-chained). In other embodiments,
R.sup.2B can be an optionally substituted --O--C.sub.3-6 alkenyl.
In some embodiments, R.sup.2B can be a substituted --O--C.sub.3-6
alkenyl. In other embodiments, R.sup.2B can be an unsubstituted
--O--C.sub.3-6 alkenyl. In still other embodiments, R.sup.2B co be
an optionally substituted --O--C.sub.3-6 alkynyl. In some
embodiments, R.sup.2B can be a substituted --O--C.sub.3-6 alkynyl.
In other embodiments, R.sup.2B can be an unsubstituted
--O--C.sub.3-6 alkynyl. In yet still other embodiments, R.sup.2B
can be cyano.
[0143] Variety of substituents can be present at the 2'-position of
the pentose ring. In some embodiments, R.sup.4B can be hydrogen. In
other embodiments, R.sup.4B can be halogen, such as fluoro. In some
embodiments, R.sup.3B can be halogen, such as fluoro. In some
embodiments, R.sup.4B can be hydrogen and R.sup.3B can be halogen.
In other embodiments, R.sup.3B and R.sup.4B can be both halogen.
For example, R.sup.3B and R.sup.4B can be both fluoro.
[0144] In some embodiments, Z.sup.1B can be O (oxygen). In other
embodiments, Z.sup.1B can be S (sulfur).
[0145] Various optionally substituted heterocyclic bases can be
attached to the pentose ring. In some embodiments, one or more of
the amine and/or amino groups may be protected with a suitable
protecting group. For example, an amino group may be protected by
transforming the amine and/or amino group to an amide or a
carbamate. In some embodiments, an optionally substituted
heterocyclic base or an optionally substituted heterocyclic base
with one or more protected amino groups can have one of the
following structures:
##STR00088##
wherein: R.sup.AB2 can be selected from hydrogen, halogen and
NHR.sup.JB2, wherein R.sup.JB2 can be selected from hydrogen,
--C(.dbd.O)R.sup.KB2 and --C(.dbd.O)OR.sup.LB2; R.sup.BB2 can be
halogen or NHR.sup.WB2, wherein R.sub.WB2 can be selected from
hydrogen, an optionally substituted C.sub.1-6 alkyl, an optionally
substituted C.sub.2-6 alkenyl, an optionally substituted C.sub.3-8
cycloalkyl, --C(.dbd.O)R.sup.MB2 and --C(.dbd.O)OR.sup.NB2;
R.sup.CB2 can be hydrogen or NHR.sup.OB2, wherein R.sup.OB2 can be
selected from hydrogen, --C(.dbd.O)R.sup.PB2 and
--C(.dbd.O)OR.sup.QB2; R.sup.DB2 can be selected from hydrogen,
halogen, an optionally substituted C.sub.1-6 alkyl, an optionally
substituted C.sub.2-6 alkenyl and an optionally substituted
C.sub.2-6 alkynyl; R.sup.EB2 can be selected from hydrogen,
hydroxy, an optionally substituted C.sub.1-6 alkyl, an optionally
substituted C.sub.3-8 cycloalkyl, --C(.dbd.O)R.sup.RB2 and
--C(.dbd.O)OR.sup.SB2; R.sup.FB2 can be selected from hydrogen,
halogen, an optionally substituted C.sub.1-6 alkyl, an optionally
substituted C.sub.2-6 alkenyl and an optionally substituted
C.sub.2-6 alkynyl; Y.sup.2B and Y.sup.3B can be independently N
(nitrogen) or CR.sup.IB2, wherein R.sup.IB2 can be selected from
hydrogen, halogen, an optionally substituted C.sub.1-6-alkyl, an
optionally substituted C.sub.2-6-alkenyl and an optionally
substituted C.sub.2-6-alkynyl; R.sup.GB2 can be an optionally
substituted C.sub.1-6 alkyl; R.sup.HB2 can be hydrogen or
NHR.sup.TB2, wherein R.sup.TB2 can be independently selected from
hydrogen, --C(.dbd.O)R.sup.UB2 and C(.dbd.O)OR.sup.VB2; and
R.sub.KB2, R.sup.LB2, R.sup.MB2, R.sup.NB2, R.sup.PB2, R.sup.QB2,
R.sub.RB2, R.sup.SB2, R.sup.UB2 and R.sup.VB2 can be independently
selected from C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl, C.sub.3-6 cycloalkyl, C.sub.3-6 cycloalkenyl, C.sub.6-10
aryl, heteroaryl, heteroalicyclyl, aryl(C.sub.1-6 alkyl),
heteroaryl(C.sub.1-6 alkyl) and heteroalicyclyl(C.sub.1-6 alkyl).
In some embodiments, the structures shown above can be modified by
replacing one or more hydrogens with substituents selected from the
list of substituents provided for the definition of
"substituted."
[0146] In some embodiments, B.sup.1B can be
##STR00089##
In other embodiments, B.sup.1B can be
##STR00090##
In still other embodiments, B.sup.1B can be
##STR00091##
such as
##STR00092##
In yet still other embodiments, B.sup.1B can be
##STR00093##
for example,
##STR00094##
In some embodiments, R.sup.DB2 can be hydrogen. In other
embodiments, B.sup.1B can be
##STR00095##
In some embodiments, R.sup.BB2 can be NH.sub.2. In other
embodiments, R.sup.BB2 can be NHR.sup.WB2, wherein R.sup.WB2 can be
--C(.dbd.O)R.sup.MB2 or --C(.dbd.O)OR.sup.NB2. In still other
embodiments, B.sup.1B can be
##STR00096##
In some embodiments, B.sup.1B can be
##STR00097##
[0147] In some embodiments, a compound of Formula (II) can have the
following structure:
##STR00098##
or a pharmaceutically acceptable salt of the foregoing. In some
embodiments of this paragraph, B.sup.1B can be an optionally
substituted purine base. In other embodiments of this paragraph,
B.sup.1B can be an optionally substituted pyrimidine base. In some
embodiments of this paragraph, B.sup.1B can be guanine. In other
embodiments of this paragraph, B.sup.1B can be thymine. In still
other embodiments of this paragraph, B.sup.1B can be cytosine. In
yet still other embodiments of this paragraph, B.sup.1B can be
uracil. In some embodiments of this paragraph, B.sup.1B can be
adenine. In some embodiments of this paragraph, Z.sup.1B can be
oxygen. In some embodiments of this paragraph, Z.sup.1B can be
sulfur. In still other embodiments of this paragraph, R.sup.1B can
be alkylcarbonyloxyalkoxy.
[0148] In some embodiments, the compound can be a compound of
Formula (III), or a pharmaceutically acceptable salt thereof,
wherein: B.sup.1C can be an optionally substituted heterocyclic
base or an optionally substituted heterocyclic base with a
protected amino group; R.sup.1C and R.sup.2C can be independently
selected from O.sup.-, OH, an optionally substituted C.sub.1-6
alkoxy,
##STR00099##
an optionally substituted N-linked amino acid and an optionally
substituted N-linked amino acid ester derivative; R.sup.3C can be
selected from an optionally substituted C.sub.1-6 alkyl, an
optionally substituted C.sub.2-6 alkenyl, an optionally substituted
C.sub.2-6 alkynyl, an optionally substituted --O--C.sub.1-6 alkyl,
an optionally substituted --O--C.sub.3-6 alkenyl, an optionally
substituted --O--C.sub.3-6 alkynyl, an optionally substituted
C.sub.3-6 cycloalkyl and cyano; R.sup.4 can be selected from OH,
--OC(.dbd.O)R.sup.''C and an optionally substituted O-linked amino
acid; R.sup.5 can be a halogen; R.sup.6 can be hydrogen or halogen;
R.sup.9C, R.sup.10C, R.sup.12C and R.sup.13C can be independently
selected from hydrogen, an optionally substituted C.sub.1-24 alkyl
and an optionally substituted aryl; R.sup.''C and R.sup.14 can be
independently selected from hydrogen, an optionally substituted
C.sub.1-24 alkyl, an optionally substituted aryl, an optionally
substituted --O--C.sub.1-24 alkyl and an optionally substituted
--O-aryl; R.sup.5 can be selected from hydrogen, an optionally
substituted C.sub.1-24 alkyl and an optionally substituted aryl;
can be a single bond or a double bond; when is a single bond, each
R.sup.7C and each R.sup.8C can be independently hydrogen or
halogen; and when is a double bond, each R.sup.7C is absent and
each R.sup.8C can be independently hydrogen or halogen; Z.sup.1C
can be O (oxygen) or S (sulfur); and R.sup.''C can be an optionally
substituted C.sub.1-24-alkyl.
[0149] In some embodiments, can be a single bond such that Formula
(III) has the structure
##STR00100##
wherein each R.sup.7C and each R.sup.8C can be independently
hydrogen or halogen. In some embodiments, the R.sup.7C and the
R.sup.8C groups can all be hydrogen. In other embodiments, one
R.sup.7C can be halogen, one R.sup.7C can be hydrogen and both
R.sup.8C groups can all be hydrogen. In still other embodiments,
one R.sup.7C can be halogen, one R.sup.7C can be hydrogen, one
R.sup.8C can be halogen and one R.sup.8C can be hydrogen. In some
embodiments, the carbon adjacent to the phosphorus and the
5'-carbon can each be independently a (S)-chiral center. In some
embodiments, the carbon adjacent to the phosphorus and the
5'-carbon can each be independent a (R)-chiral center.
[0150] In some embodiments, can be a double bond such that Formula
(III) has the structure
##STR00101##
wherein each R.sup.7C is absent and each R.sup.8C can be
independently hydrogen or halogen. In some embodiments, both
R.sup.8C groups can be hydrogen. In other embodiments, one R.sup.8C
can be halogen and the other R.sup.8C can be hydrogen. In some
embodiments, both R.sup.8C groups can be halogen. In some
embodiments, the double bond has a (Z)-configuration. In some
embodiments, the double bond has a (E)-configuration.
[0151] In some embodiments, R.sup.1C and/or R.sup.2C can be
O.sup.-. In other embodiments, R.sup.1C and/or R.sup.2C can be OH.
In some embodiments, R.sup.1C and R.sup.2C can be both OH.
[0152] In some embodiments, R.sup.1C and/or R.sup.2C can be
##STR00102##
wherein R.sup.9C and R.sup.10C can be independently selected from
hydrogen, an optionally substituted C.sub.1-24 alkyl and an
optionally substituted aryl; and R.sup.11C can be selected from
hydrogen, an optionally substituted C.sub.1-24 alkyl, an optionally
substituted aryl, an optionally substituted --O--C.sub.1-24 alkyl
and an optionally substituted --O-aryl. In some embodiments,
R.sup.9C and R.sup.10C can be hydrogen. In other embodiments, at
least one of R.sup.9C and R.sup.10C can be an optionally
substituted C.sub.1-24 alkyl or an optionally substituted aryl. In
some embodiments, R.sup.11C can be an optionally substituted
C.sub.1-24 alkyl. In other embodiments, R.sup.11C can be an
optionally substituted aryl. In still other embodiments, R.sup.11C
can be an optionally substituted --O--C.sub.1-24 alkyl or an
optionally substituted --O-aryl. In some embodiments, R.sup.1C and
R.sup.2C can be both
##STR00103##
[0153] In some embodiments, R.sup.1C and/or R.sup.2C can be
##STR00104##
wherein R.sup.12C and R.sup.13C can be independently selected from
hydrogen, an optionally substituted C.sub.1-24 alkyl and an
optionally substituted aryl; R.sup.14C can be independently
selected from hydrogen, an optionally substituted C.sub.1-24 alkyl,
an optionally substituted aryl, an optionally substituted
--O--C.sub.1-24 alkyl and an optionally substituted --O-aryl; and
Z.sup.1C can be independently O (oxygen) or S (sulfur). In some
embodiments, R.sup.12C and R.sup.13C can be hydrogen. In other
embodiments, at least one of R.sup.12C and R.sup.13C can be an
optionally substituted C.sub.1-24 alkyl or an optionally
substituted aryl. In some embodiments, R.sup.14C can be an
optionally substituted C.sub.1-24 alkyl. In other embodiments,
R.sup.14C can be an optionally substituted aryl. In still other
embodiments, R.sup.14C can be an optionally substituted
--O--C.sub.1-24 alkyl or an optionally substituted --O-aryl. In
some embodiments, Z.sup.1C can be O (oxygen). In other embodiments,
Z.sup.1C can be or S (sulfur). In some embodiments, R.sup.1C and/or
R.sup.2C can be isopropylcarbonyloxymethoxy. In some embodiments,
R.sup.1C and/or R.sup.2C can be pivaloyloxymethoxy. In some
embodiments, R.sup.1C and R.sup.2C can be both
##STR00105##
In some embodiments, R.sup.1C and R.sup.2C can be both
isopropylcarbonyloxymethoxy. In other embodiments, R.sup.1C and
R.sup.2C can be both pivaloyloxymethoxy.
[0154] In some embodiments, R.sup.1C and/or R.sup.2C can be
##STR00106##
In some embodiments, R.sup.15C can be hydrogen. In other
embodiments, R.sup.15C can be an optionally substituted C.sub.1-24
alkyl. In still other embodiments, R.sup.15C can be an optionally
substituted aryl. In some embodiments, R.sup.15C can be a C.sub.1-6
alkyl, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl,
isobutyl, tert-butyl, pentyl (branched and straight-chained), and
hexyl (branched and straight-chained). In some embodiments,
R.sup.1C and R.sup.2C can be both
##STR00107##
[0155] In some embodiments, R.sup.1C and/or R.sup.2C can be an
optionally substituted N-linked amino acid or an optionally
substituted N-linked amino acid ester derivative. For example,
R.sup.1C and/or R.sup.2C can be optionally substituted version of
the following: alanine, asparagine, aspartate, cysteine, glutamate,
glutamine, glycine, proline, serine, tyrosine, arginine, histidine,
isoleucine, leucine, lysine, methionine, phenylalanine, threonine,
tryptophan, valine and ester derivatives thereof. In some
embodiments, R.sup.1C and/or R.sup.2C can be selected from alanine
isopropyl ester, alanine cyclohexyl ester, alanine neopentyl ester,
valine isopropyl ester and leucine isopropyl ester. In some
embodiments, R.sup.1C and/or R.sup.2C can have the structure
##STR00108##
wherein R.sup.19C can be selected from hydrogen, an optionally
substituted C.sub.1-6-alkyl, an optionally substituted C.sub.3-6
cycloalkyl, an optionally substituted aryl, an optionally
substituted aryl(C.sub.1-6 alkyl) and an optionally substituted
haloalkyl; R.sup.20C can be selected from hydrogen, an optionally
substituted C.sub.1-6 alkyl, an optionally substituted C.sub.1-6
haloalkyl, an optionally substituted C.sub.3-6 cycloalkyl, an
optionally substituted C.sub.6 aryl, an optionally substituted
C.sub.10 aryl and an optionally substituted aryl(C.sub.1-6 alkyl);
and R.sup.21C can be hydrogen or an optionally substituted
C.sub.1-4-alkyl; or R.sup.20C and R.sup.21C can be taken together
to form an optionally substituted C.sub.3-6 cycloalkyl.
[0156] When R.sup.20C is substituted, R.sup.20C can be substituted
with one or more substituents selected from N-amido, mercapto,
alkylthio, an optionally substituted aryl, hydroxy, an optionally
substituted heteroaryl, O-carboxy, and amino. In some embodiments,
R.sup.20C can be an unsubstituted C.sub.1-6-alkyl, such as those
described herein. In some embodiments, R.sup.20C can be hydrogen.
In other embodiments, R.sup.20C can be methyl. In some embodiments,
R.sup.19C can be an optionally substituted C.sub.1-6 alkyl.
Examples of optionally substituted C.sub.1-6-alkyls include
optionally substituted variants of the following: methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl
(branched and straight-chained), and hexyl (branched and
straight-chained). In some embodiments, R.sup.19C can be methyl or
isopropyl. In some embodiments, R.sup.19C can be ethyl or
neopentyl. In other embodiments, R.sup.19C can be an optionally
substituted C.sub.3-6 cycloalkyl. Examples of optionally
substituted C.sub.3-6 cycloalkyl include optionally substituted
variants of the following: cyclopropyl, cyclobutyl, cyclopentyl,
and cyclohexyl. In an embodiment, R.sup.19C can be an optionally
substituted cyclohexyl. In still other embodiments, R.sup.19C can
be an optionally substituted aryl, such as phenyl and naphthyl. In
yet still other embodiments, R.sup.19C can be an optionally
substituted aryl(C.sub.1-6 alkyl). In some embodiments, R.sup.19C
can be an optionally substituted benzyl. In some embodiments,
R.sup.19C can be an optionally substituted C.sub.1-6 haloalkyl, for
example, CF.sub.3. In some embodiments, R.sup.21C can be hydrogen.
In other embodiments, R.sup.21C can be an optionally substituted
C.sub.1-4-alkyl, such as methyl, ethyl, n-propyl, isopropyl,
n-butyl, isobutyl and tert-butyl. In an embodiment, R.sup.21C can
be methyl. In some embodiments, R.sup.20C and R.sup.21C can be
taken together to form an optionally substituted C.sub.3-6
cycloalkyl. Examples of optionally substituted C.sub.3-6 cycloalkyl
include optionally substituted variants of the following:
cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Depending on
the groups that are selected for R.sup.20C and R.sup.21C, the
carbon to which R.sup.20C and R.sup.21C are attached may be a
chiral center. In some embodiment, the carbon to which R.sup.20C
and R.sup.21C are attached may be a (R)-chiral center. In other
embodiments, the carbon to which R.sup.20C and R.sup.21C are
attached may be a (S)-chiral center.
[0157] Examples of suitable
##STR00109##
groups include the following:
##STR00110## ##STR00111##
[0158] In some embodiments, R.sup.1C and R.sup.2C can be the same.
In other embodiments, R.sup.1C and R.sup.2C can be different.
[0159] In some embodiments, R.sup.1C can be
##STR00112##
and R.sup.2C can be O.sup.- or OH, wherein R.sup.16C, R.sup.17C and
R.sup.18C can be absent or hydrogen; and n can be 0 or 1. Those
skilled in the art understand that when R.sup.16C, R.sup.17C and/or
R.sup.18C are absent, the associated oxygen will be negatively
charge. In some embodiments, when n is 0, the compound of Formula
(III) will be a diphosphate. In other embodiments, when n is 1, the
compound of Formula (III) will be a triphosphate.
[0160] A variety of substituents can be present at the 4'-position
of the pentose ring. In some embodiments, R.sup.3C can be an
optionally substituted C.sub.1-6 alkyl. Examples of suitable
C.sub.1-6 alkyls include methyl, ethyl, n-propyl, isopropyl,
n-butyl, isobutyl, tert-butyl, pentyl (branched and
straight-chained), and hexyl (branched and straight-chained). In
some embodiments, R.sup.3C can be an unsubstituted C.sub.1-6 alkyl.
In other embodiments, R.sup.3C can be a substituted C.sub.1-6
alkyl. For example, R.sup.3C can be a halogen substituted C.sub.1-6
alkyl. In other embodiments, R.sup.3C can be an optionally
substituted C.sub.2-6 alkenyl. In some embodiments, R.sup.3C can be
a substituted C.sub.2-6 alkenyl. In other embodiments, R.sup.3C can
be an unsubstituted C.sub.2-6 alkenyl. For example, R.sup.3C can be
ethenyl, propenyl or allenyl. In still other embodiments, R.sup.3C
can be an optionally substituted C.sub.2-6 alkynyl. In some
embodiments, R.sup.3 can be a substituted C.sub.2-6 alkynyl. In
other embodiments, R.sup.3C can be an unsubstituted C alkynyl.
Suitable C.sub.2-6 alkynyls include ethynyl and propynyl. In yet
still other embodiments, R.sup.3C can be an optionally substituted
C.sub.3-6 cycloalkyl. In some embodiments, R.sup.3C can be a
substituted C.sub.3-6 cycloalkyl. In other embodiments, R.sup.3C
can be an unsubstituted C.sub.3-6 cycloalkyl. A non-limiting list
of C.sub.3-6 cycloalkyls include cyclopropyl, cyclobutyl,
cyclopentyl and cyclohexyl. In some embodiments, R.sup.3 can be an
optionally substituted --O--C.sub.1-6 alkyl. In some embodiments,
R.sup.3C can be a substituted --O--C.sub.1-6 alkyl. In other
embodiments, R.sup.3C can be an unsubstituted --O--C.sub.1-6 alkyl.
Examples of suitable O--C.sub.1-6 alkyl groups include methoxy,
ethoxy, n-propoxy, iso-propoxy, n-butoxy, isobutoxy, tert-butoxy,
pentoxy (branched and straight-chained), and hexoxy (branched and
straight-chained). In other embodiments, R.sup.3C can be an
optionally substituted --O--C.sub.3-6 alkenyl. In some embodiments,
R.sup.3C can be a substituted --O--C.sub.3 alkenyl. In other
embodiments, R.sup.3C can be an unsubstituted --O--C.sub.3-6
alkenyl. In still other embodiments, R.sup.3C can be an optionally
substituted --O--C.sub.3-6 alkynyl. In some embodiments, R.sup.3C
can be a substituted --O--C.sub.3 alkynyl. In other embodiments,
R.sup.3C can be an unsubstituted --O--C.sub.3-6 alkynyl. In yet
still other embodiments, R.sup.3C can be cyano.
[0161] The substituents that can be present on the 3'-position of
the pentose ring can vary. In some embodiments, R.sup.4C can be OH.
In other embodiments, R.sup.4C can be an optionally substituted
O-linked amino acid. Examples of suitable O-linked amino acids
include alanine, asparagine, aspartate, cysteine, glutamate,
glutamine, glycine, proline, serine, tyrosine, arginine, histidine,
isoleucine, leucine, lysine, methionine, phenylalanine, threonine,
tryptophan and valine. Additional examples of suitable amino acids
include, but are not limited to, ornithine, hypusine,
2-aminoisobutyric acid, dehydroalanine, gamma-aminobutyric acid,
citrulline, beta-alanine, alpha-ethyl-glycine, alpha-propyl-glycine
and norleucine. In some embodiments, the O-linked amino acid can
have the structure
##STR00113##
wherein R.sup.22C can be selected from hydrogen, an optionally
substituted C.sub.1-6 alkyl, an optionally substituted C.sub.1-6
haloalkyl, an optionally substituted C.sub.3-6 cycloalkyl, an
optionally substituted C.sub.6 aryl, an optionally substituted
C.sub.10 aryl and an optionally substituted aryl(C.sub.1-6 alkyl);
and R.sup.23C can be hydrogen or an optionally substituted
C.sub.1-4-alkyl; or R.sup.22C and R.sup.23C can be taken together
to form an optionally substituted C.sub.3-6 cycloalkyl.
[0162] When R.sup.22C is substituted, R.sup.22C can be substituted
with one or more substituents selected from N-amido, mercapto,
alkylthio, an optionally substituted aryl, hydroxy, an optionally
substituted heteroaryl, O-carboxy, and amino. In some embodiments,
R.sup.22C can be an unsubstituted C.sub.1-6-alkyl, such as those
described herein. In some embodiments, R.sup.22C can be hydrogen.
In other embodiments, R.sup.22C can be methyl. In some embodiments,
R.sup.23C can be hydrogen. In other embodiments, R.sup.23C can be
an optionally substituted C.sub.1-4-alkyl, such as methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl. In an
embodiment, R.sup.23C can be methyl. Depending on the groups that
are selected for R.sup.22C and R.sup.23C, the carbon to which
R.sup.22C and R.sup.23C are attached may be a chiral center. In
some embodiment, the carbon to which R.sup.22C and R.sup.23C are
attached may be a (R)-chiral center. In other embodiments, the
carbon to which R.sup.22C and R.sup.23C are attached may be a
(S)-chiral center.
[0163] Examples of suitable
##STR00114##
include the following:
##STR00115##
[0164] In still other embodiments, R.sup.4C can be
--OC(.dbd.O)R.sup.''C, wherein R.sup.''C can be an optionally
substituted C.sub.1-24 alkyl. In some embodiments, R.sup.''C can be
a substituted C.sub.1-12 alkyl. In other embodiments, R.sup.''C can
be an unsubstituted C.sub.1-12 alkyl. In still other embodiments,
R.sup.''C can be a substituted C.sub.1-8 alkyl. In yet still other
embodiments, R.sup.''C can be an unsubstituted C.sub.1-8 alkyl. In
some embodiments, R.sup.4C can be an optionally substituted acyl.
In other embodiments, R.sup.4C can be --OC(.dbd.O)RT, wherein
R.sup.''C can be selected from an optionally substituted C.sub.1-12
alkyl, an optionally substituted C.sub.2-12 alkenyl, an optionally
substituted C.sub.2-12 alkynyl, an optionally substituted C.sub.3-8
cycloalkyl, an optionally substituted C.sub.5-8 cycloalkenyl, an
optionally substituted C.sub.6-10 aryl, an optionally substituted
heteroaryl, an optionally substituted heterocyclyl, an optionally
substituted aryl(C.sub.1-6 alkyl), an optionally substituted
heteroaryl(C.sub.1-6 alkyl) and an optionally substituted
heterocyclyl(C.sub.1-6 alkyl). In some embodiments, R.sup.''C can
be a substituted C.sub.1-12 alkyl. In other embodiments, R.sup.''C
can be an unsubstituted C.sub.1-12 alkyl.
[0165] A variety of substituents can also be present at the
2'-position of the pentose ring. In some embodiments, R.sup.6C can
be hydrogen. In other embodiments, R.sup.6C can be halogen, such as
fluoro. In some embodiments, R.sup.5C can be halogen, such as
fluoro. In some embodiments, R.sup.6C can be hydrogen and R.sup.5C
can be halogen. In other embodiments, R.sup.5C and R.sup.6C can be
both halogen. For example, R.sup.5C and R.sup.6C can be both
fluoro.
[0166] Various optionally substituted heterocyclic bases can be
attached to the pentose ring. In some embodiments, one or more of
the amine and/or amino groups may be protected with a suitable
protecting group. For example, an amino group may be protected by
transforming the amine and/or amino group to an amide or a
carbamate. In some embodiments, an optionally substituted
heterocyclic base or an optionally substituted heterocyclic base
with one or more protected amino groups can have one of the
following structures:
##STR00116##
wherein: R.sup.AC2 can be selected from hydrogen, halogen and
NHR.sup.JC2, wherein R.sup.JC2 can be selected from hydrogen,
--C(.dbd.O)R.sup.KC2 and --C(.dbd.O)OR.sup.LC2; R.sup.BC2 can be
halogen or NHR.sup.WC2, wherein R.sub.WC2 can be selected from
hydrogen, an optionally substituted C.sub.1-6 alkyl, an optionally
substituted C.sub.2-6 alkenyl, an optionally substituted C.sub.3-8
cycloalkyl, --C(.dbd.O)R.sup.MC2 and --C(.dbd.O)OR.sup.NC2;
R.sup.CC2 can be hydrogen or NHR.sup.OC2, wherein R.sub.OC2 can be
selected from hydrogen, --C(.dbd.O)R.sup.PC2 and
--C(.dbd.O)OR.sup.QC2; R.sup.DC2 can be selected from hydrogen,
halogen, an optionally substituted C.sub.1-6 alkyl, an optionally
substituted C.sub.2-6 alkenyl and an optionally substituted
C.sub.2-6 alkynyl; R.sup.EC2 can be selected from hydrogen,
hydroxy, an optionally substituted C.sub.1-6 alkyl, an optionally
substituted C.sub.3-8 cycloalkyl, --C(.dbd.O)R.sup.RC2 and
--C(.dbd.O)OR.sup.SC2; R.sup.FC2 can be selected from hydrogen,
halogen, an optionally substituted C.sub.1-6 alkyl, an optionally
substituted C.sub.2-6 alkenyl and an optionally substituted
C.sub.2-6 alkynyl; Y.sup.2C and Y.sup.3C can be independently N
(nitrogen) or CR.sup.IC2, wherein R.sup.IC2 can be selected from
hydrogen, halogen, an optionally substituted C.sub.1-6-alkyl, an
optionally substituted C.sub.2-6-alkenyl and an optionally
substituted C.sub.2-6-alkynyl; R.sup.GC2 can be an optionally
substituted C.sub.1-6 alkyl; R.sup.HC2 can be hydrogen or
NHR.sup.TC2, wherein R.sup.TC2 can be independently selected from
hydrogen, --C(.dbd.O)R.sub.UC2 and --C(.dbd.O)OR.sup.VC2; and
R.sup.KC2, R.sup.LC2, R.sup.MC2, R.sup.NC2, R.sup.PC2, R.sup.QC2,
R.sup.RC2, R.sup.SC2, R.sup.UC2 and R.sup.VC2 can be independently
selected from C.sub.1-6 alkyl, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl, C.sub.3-6 cycloalkyl, C.sub.3-6 cycloalkenyl, C.sub.6-10
aryl, heteroaryl, heteroalicyclyl, aryl(C.sub.1-6 alkyl),
heteroaryl(C.sub.1-6 alkyl) and heteroalicyclyl(C.sub.1-6 alkyl).
In some embodiments, the structures shown above can be modified by
replacing one or more hydrogens with substituents selected from the
list of substituents provided for the definition of
"substituted."
[0167] In some embodiments, B.sup.1C can be
##STR00117##
In other embodiments, B.sup.1C can be
##STR00118##
In still other embodiments, B.sup.1C can be
##STR00119##
such as
##STR00120##
In yet still other embodiments, B.sup.1C can be
##STR00121##
for example,
##STR00122##
In some embodiments, R.sup.DC2 can be hydrogen. In other
embodiments, B.sup.1C can be
##STR00123##
In some embodiments, R.sup.BC2 can be NH.sub.2. In other
embodiments, R.sup.BC2 can be NHR.sup.WC2, wherein R.sup.WC2 can be
--C(.dbd.O)R.sup.MC2 or --C(.dbd.O)OR.sup.NC2. In still other
embodiments, B.sup.1C can be
##STR00124##
In some embodiments, B.sup.1C can be
##STR00125##
[0168] In some embodiments, the compound of Formula (III) can have
one of the following structures:
##STR00126##
In some embodiments of this paragraph, B.sup.1C can be an
optionally substituted purine base. In other embodiments of this
paragraph, B.sup.1C can be an optionally substituted pyrimidine
base. In some embodiments of this paragraph, B.sup.1C can be
guanine. In other embodiments of this paragraph, B.sup.1C can be
thymine. In still other embodiments of this paragraph, B.sup.1C can
be cytosine. In yet still other embodiments of this paragraph,
B.sup.1C can be uracil. In some embodiments of this paragraph,
B.sup.1C can be adenine. In some embodiments of this paragraph,
R.sup.1C and R.sup.2C can each be an optionally substituted
C.sub.1-4 alkyl. In other embodiments of this paragraph, R.sup.1A
can be an optionally substituted acyl. In still other embodiments
of this paragraph, R.sup.1C and R.sup.2C can form a mono-, di- or
tri-phosphate. In yet other embodiments of this paragraph, R.sup.1C
and R.sup.2C can each be an alkylcarbonyloxyalkoxy. In some
embodiments of this paragraph, R.sup.4C can be OH. In some
embodiments of this paragraph, R.sup.5C can be F and R.sup.6C can
be hydrogen.
[0169] Examples of suitable compounds of Formula (I) include, but
are not limited to the following:
##STR00127## ##STR00128## ##STR00129## ##STR00130## ##STR00131##
##STR00132## ##STR00133## ##STR00134##
or a pharmaceutically acceptable salt of the foregoing.
[0170] Additional examples of a compound of Formula (I) include the
following:
##STR00135## ##STR00136## ##STR00137## ##STR00138## ##STR00139##
##STR00140## ##STR00141## ##STR00142##
or a pharmaceutically acceptable salt of the foregoing.
[0171] Further examples of a compound of Formula (I) include, but
are not limited to the following:
##STR00143## ##STR00144## ##STR00145## ##STR00146## ##STR00147##
##STR00148## ##STR00149##
or a pharmaceutically acceptable salt of the foregoing.
[0172] Examples of a compound of Formula (II) include, but are not
limited to, the following:
##STR00150##
or a pharmaceutically acceptable salt of the foregoing.
[0173] Examples of a compound of Formula (III) include, but are not
limited to, the following:
##STR00151## ##STR00152##
or a pharmaceutically acceptable salt of the foregoing.
[0174] Further examples of a compound of Formula (III) include, but
are not limited to, the following:
##STR00153##
or a pharmaceutically acceptable salt of the foregoing.
Synthesis
[0175] Compounds of Formula (I) Formula (II) and Formula (III), and
those described herein may be prepared in various ways. Some
compounds of Formulae (I), (II) and (III) can be obtained
commercially and/or prepared utilizing known synthetic procedures.
General synthetic routes to the compounds of Formulae (I), (II) and
(III), and some examples of starting materials used to synthesize
the compounds of Formulae (I), (II) and (III) are shown and
described herein. The routes shown and described herein are
illustrative only and are not intended, nor are they to be
construed, to limit the scope of the claims in any manner
whatsoever. Those skilled in the art will be able to recognize
modifications of the disclosed syntheses and to devise alternate
routes based on the disclosures herein; all such modifications and
alternate routes are within the scope of the claims.
##STR00154##
[0176] As shown in Scheme 1, compounds of Formula (I) can be
prepared from a nucleoside, for example, a nucleoside of Formula
(A). In Scheme 1, R.sup.3a, R.sup.4a, R.sup.5a, and B.sup.1a can be
the same as R.sup.3A, R.sup.4A, R.sup.5A, and B.sup.1A as described
herein for Formula (I), and PG.sup.1 is a suitable protecting
group. A hydroxyalkyl group can be formed at the 4'-position of the
pentose ring using suitable conditions known to those skilled in
the art. Examples of suitable conditions for forming a hydroxyalkyl
include the use of 2-iodoxybenzoic acid (IBX) aqueous formaldehyde
and sodium borohydride. A compound of Formula (B) can be oxidized
to an aldehyde using a suitable oxidizing agent(s) to form a
compound of Formula (C). An example of suitable oxidizing agent is
Dess-Martin periodinane. An optionally substituted C.sub.2-6
alkenyl or an optionally substituted C.sub.2-6 alkynyl can be
formed at the 4'-position using methods known to those skilled in
the art, for example, Wittig reagent and n-BuLi, Wittig-type
reactions, Peterson olefination reaction, and Corey Fuchs reaction.
An optionally substituted C.sub.1-6 alkyl can be obtained by
hydrogenating the unsaturated group attached to the 4'-position,
for example, using hydrogen over palladium on carbon.
[0177] Alternatively, a compound of Formula (B) can be transformed
to a haloalkyl using a suitable agent(s), for example, to an iodide
using imidazole, triphenylphosphine and iodine; to a fluoro using
diethylaminosulfur trifluoride (DAST); or to a chloro using
triphenylphosphine and carbontetrachloride in dichloroethylene
(DCE). An iodoalkyl can be transformed to an unsubstituted
C.sub.1-6 alkyl group using methods known to those skilled in the
art, for example, hydrogen over palladium on carbon. A compound of
Formula (C) can be reacted with hydroxylamine to form an oxime. The
oxime can be transformed to a cyano group using methods known to
those skilled in the art, for example, using methanesulfonyl
chloride.
##STR00155##
[0178] As shown in Scheme 2, compounds of Formula (I), where
R.sup.2A is an optionally substituted --O--C.sub.1-6 alkyl, an
optionally substituted --O--C.sub.3-6 alkenyl or an optionally
substituted --O--C.sub.3-6 alkynyl, can be prepared from a
nucleoside, for example, a nucleoside of Formula (A). In Scheme 2,
R.sup.2a, R.sup.3a, R.sup.4a, R.sup.5a and B.sup.1a can be the same
as R.sup.2A, R.sup.3A, R.sup.4A, R.sup.5A and B.sup.1A as described
herein for Formula (I), and PG.sup.2 can be a suitable protecting
group. The nucleoside can undergo elimination and form an olefin
having the general formula of Formula (D). A compound of Formula
(D) can be treated with an iodinating reagent in the presence of
lead carbonate and an alkoxy source to form a compound of Formula
(E). A compound of Formula (E) can then be transformed to a
compound of Formula (I) through displacement of the iodide with an
oxygen nucleophile.
##STR00156##
##STR00157##
[0179] Compounds of Formula (I) having a phosphorus containing
group attached to the 5'-position of the pentose ring can be
prepared using various methods known to those skilled in the art.
Examples of methods are shown in Schemes 3 and 4. A phosphorus
containing precursor can be coupled to the nucleoside, for example,
a compound of Formula (F) or a compound of Formula (G). As shown in
Scheme 3, following the coupling of the phosphorus containing
precursor, any leaving groups can be cleaved under suitable
conditions, such as hydrolysis. Further phosphorus containing
groups can be added using methods known to those skilled in the
art, for example using a pyrophosphate.
[0180] In some embodiments, an alkoxide can be generated from a
compound of Formula (G) using an organometallic reagent, such as a
Grignard reagent. The alkoxide can be coupled to the phosphorus
containing precursor. Suitable Grignard reagents are known to those
skilled in the art and include, but are not limited to,
alkylmagnesium chlorides and alkylmagnesium bromides. In some
embodiments, an appropriate base can be used. Examples of suitable
bases include, but are not limited to, an amine base, such as an
alkylamine (including mono-, di- and tri-alkylamines (e.g.,
triethylamine)), optionally substituted pyridines (e.g. collidine)
and optionally substituted imidazoles (e.g., N-methylimidazole)).
Alternatively, a phosphorus containing precursor can be added to
the nucleoside and form a phosphite. The phosphite can be oxidized
to a phosphate using conditions known to those skilled in the art.
Suitable conditions include, but are not limited to,
meta-chloroperoxybenzoic acid (MCPBA) and iodine as the oxidizing
agent and water as the oxygen donor.
[0181] When compounds of Formula (I) have Z.sup.1A, Z.sup.2A or
Z.sup.3A being sulfur, the sulfur can be added in various manners
known to those skilled in the art. In some embodiments, the sulfur
can be part of the phosphorus containing precursor, for
example,
##STR00158##
Alternatively, the sulfur can be added using a sulfurization
reagent. Suitable sulfurization agents are known to those skilled
in the art, and include, but are not limited to, elemental sulfur,
Lawesson's reagent, cyclooctasulfur,
3H-1,2-Benzodithiole-3-one-1,1-dioxide (Beaucage's reagent),
3-((N,N-dimethylaminomethylidene)amino)-3H-1,2,4-dithiazole-5-t-
hione (DDTT) and bis(3-triethoxysilyl)propyl-tetrasulfide
(TEST).
[0182] Suitable phosphorus containing precursors can be
commercially obtained or prepared by synthetic methods known to
those skilled in the art. Examples of general structures of
phosphorus containing precursors are shown in Schemes 3 and 4.
##STR00159##
[0183] A method for forming a compound of Formula (II) is shown in
Scheme 5. In Scheme 5, R.sup.1b, R.sup.2b, R.sup.3b, R.sup.4b and
B.sup.1b can be the same as R.sup.1B, R.sup.2B, R.sup.3B, R.sup.4B
and B.sup.1B as described herein for Formula (II), each L.sup.1 can
be a halogen, a sulfonate ester or an amine (mono- or
di-substituted), and X can be oxygen or sulfur. As shown in Scheme
5, a compound having a hydroxy group attached to the 3'-carbon and
a hydroxy group attached to the 5'-carbon can be reacted with a
compound having the formula, (R.sup.1b)P(L.sup.1).sub.2, in the
presence of a base, to produce a phosphite compound. Suitable bases
are known to those skilled in the art and described herein. The
phosphorus can then be oxidized to phosphorus(V) using a suitable
oxidizing agent, to produce a compound where X is O (oxygen).
Alternatively, the phosphite compound can be reacted with a
sulfurization reagent to produce a compound where X is S (sulfur).
Suitable oxidizing and sulfurization agents are known to those
skilled in the art. For example, the oxidation can be carried out
using iodine as the oxidizing agent and water as the oxygen donor.
Suitable sulfurization agents are described herein.
##STR00160##
[0184] A method for forming a compound of Formula (III) is shown in
Scheme 6. In Scheme 6, R.sup.1c, R.sup.2c, R.sup.3c, R.sup.4c,
R.sup.5c, R.sup.6c and B.sup.1c can be the same as R.sup.1C,
R.sup.2C, R.sup.3C, R.sup.4C, R.sup.5C, R.sup.6C and B.sup.1C as
described herein for Formula (III), and R.sup.7C and R.sup.8C are
not shown. The oxygen attached to the 5'-carbon of the compound of
Formula (H) can be oxidized to a ketone using methods and reagents
known to those skilled in the art. For example, an oxidizing agent,
such as Dess-Martin periodinane, can be utilized. A
phosphorus-containing reagent can then be added to a compound of
Formula (J) in the presence of a strong base (e.g., sodium
hydride). The double bond can be hydrogenated, for example using
hydrogen gas or Pd/C, to a single bond. Additional phosphates can
be added via phosphorylation to form a di- or tri-phosphate using
suitable reagents, such as a pyrophosphate (e.g.,
tetrabutylammonium pyrophosphate).
[0185] An acyl group can be added to the 5'-position and/or the
3'-position of a compound of Formula (I) or (III) using methods
known to those skilled in the art. One suitable method is using an
anhydride in pyridine.
[0186] During the synthesis of any of the compounds described
herein, if desired, any hydroxy groups attached to the pentose
ring, and any --NH and/or NH.sub.2 groups present on the B.sup.1a,
B.sup.1b and B.sup.1c can be protected with one or more suitable
protecting groups. Suitable protecting groups are described herein.
For example, when R.sup.3a and/or R.sup.4c is a hydroxy group,
R.sup.3a and/or R.sup.4c can be protected with a triarylmethyl
group or a silyl group. Likewise, any --NH and/or NH.sub.2 groups
present on the B.sup.1a, B.sup.1b and B.sup.1c can be protected,
such as with a triarylmethyl and a silyl group(s). Examples of
triarylmethyl groups include but are not limited to, trityl,
monomethoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr),
4,4',4''-trimethoxytrityl (TMTr). 4,4',4''-tris-(benzoyloxy) trityl
(TBTr), 4,4',4''-tris (4,5-dichlorophthalimido) trityl (CPTr),
4,4',4''-tris (levulinyloxy) trityl (TLTr),
p-anisyl-1-naphthylphenylmethyl, di-o-anisyl-1-naphthylmethyl,
p-tolyldipheylmethyl, 3-(imidazolylmethyl)-4,4'-dimethoxytrityl,
9-phenylxanthen-9-yl(Pixyl), 9-(p-methoxyphenyl) xanthen-9-yl
(Mox), 4-decyloxytrityl, 4-hexadecyloxytrityl,
4,4'-dioctadecyltrityl, 9-(4-octadecyloxyphenyl)xanthen-9-yl,
1,1'-bis-(4-methoxyphenyl)-1'-pyrenylmethyl,
4,4',4''-tris-(tert-butylphenyl) methyl (TTTr) and 4,4'-di-3,
5-hexadienoxytrityl. Examples of silyl groups include, but are not
limited to, trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS),
triisopropylsilyl (TIPS), tert-butyldiphenylsilyl (TBDPS),
tri-iso-propylsilyloxymethyl and [2-(trimethylsilyl)ethoxy]methyl.
Alternatively, R.sup.3a and R.sup.4a and/or R.sup.4c and R.sup.5c
can be protected by a single achiral or chiral protecting group,
for example, by forming an orthoester, a cyclic acetal or a cyclic
ketal. Suitable orthoesters include methoxymethylene acetal,
ethoxymethylene acetal, 2-oxacyclopentylidene orthoester,
dimethoxymethylene orthoester, 1-methoxyethylidene orthoester,
1-ethoxyethylidene orthoester, methylidene orthoester, phthalide
orthoester 1,2-dimethoxyethylidene orthoester, and
alpha-methoxybenzylidene orthoester, suitable cyclic acetals
include methylene acetal, ethylidene acetal, t-butylmethylidene
acetal, 3-(benzyloxy)propyl acetal, benzylidene acetal,
3,4-dimethoxybenzylidene acetal and p-acetoxybenzylidene acetal;
and suitable cyclic ketals include 1-t-butylethylidene ketal,
1-phenylethylidene ketal, isopropylidene ketal, cyclopentylidene
ketal, cyclohexylidene ketal, cycloheptylidene ketal and
1-(4-methoxyphenyl)ethylidene ketal. Those skilled in the art will
appreciate that groups attached to the pentose ring and any --NH
and/or NH.sub.2 groups present on the B.sup.1, B.sup.1b and B.sup.1
can be protected with various protecting groups, and any protecting
groups present can be exchanged for other protecting groups. The
selection and exchange of the protecting groups is within the skill
of those of ordinary skill in the art. Any protecting group(s) can
be removed by methods known in the art, for example, with an acid
(e.g., a mineral or an organic acid), a base or a fluoride
source.
[0187] Pharmaceutical Compositions
[0188] Some embodiments described herein relates to a
pharmaceutical composition, that can include an effective amount of
one or more compounds described herein (e.g., a compound of Formula
(I), a compound of Formula (II) and/or a compound of Formula (III),
or a pharmaceutically acceptable salt of the foregoing) and a
pharmaceutically acceptable carrier, diluent, excipient or
combination thereof.
[0189] The term "pharmaceutical composition" refers to a mixture of
one or more compounds disclosed herein with other chemical
components, such as diluents or carriers. The pharmaceutical
composition facilitates administration of the compound to an
organism. Pharmaceutical compositions can also be obtained by
reacting compounds with inorganic or organic acids such as
hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,
phosphoric acid, methanesulfonic acid, ethanesulfonic acid,
p-toluenesulfonic acid, and salicylic acid. Pharmaceutical
compositions will generally be tailored to the specific intended
route of administration.
[0190] The term "physiologically acceptable" defines a carrier,
diluent or excipient that does not abrogate the biological activity
and properties of the compound.
[0191] As used herein, a "carrier" refers to a compound that
facilitates the incorporation of a compound into cells or tissues.
For example, without limitation, dimethyl sulfoxide (DMSO) is a
commonly utilized carrier that facilitates the uptake of many
organic compounds into cells or tissues of a subject.
[0192] As used herein, a "diluent" refers to an ingredient in a
pharmaceutical composition that lacks pharmacological activity but
may be pharmaceutically necessary or desirable. For example, a
diluent may be used to increase the bulk of a potent drug whose
mass is too small for manufacture and/or administration. It may
also be a liquid for the dissolution of a drug to be administered
by injection, ingestion or inhalation. A common form of diluent in
the art is a buffered aqueous solution such as, without limitation,
phosphate buffered saline that mimics the composition of human
blood.
[0193] As used herein, an "excipient" refers to an inert substance
that is added to a pharmaceutical composition to provide, without
limitation, bulk, consistency, stability, binding ability,
lubrication, disintegrating ability etc., to the composition. A
"diluent" is a type of excipient.
[0194] The pharmaceutical compositions described herein can be
administered to a human patient per se, or in pharmaceutical
compositions where they are mixed with other active ingredients, as
in combination therapy, or carriers, diluents, excipients or
combinations thereof. Proper formulation is dependent upon the
route of administration chosen. Techniques for formulation and
administration of the compounds described herein are known to those
skilled in the art.
[0195] The pharmaceutical compositions disclosed herein may be
manufactured in a manner that is itself known, e.g., by means of
conventional mixing, dissolving, granulating, dragee-making,
levigating, emulsifying, encapsulating, entrapping or tableting
processes. Additionally, the active ingredients are contained in an
amount effective to achieve its intended purpose. Many of the
compounds used in the pharmaceutical combinations disclosed herein
may be provided as salts with pharmaceutically compatible
counterions.
[0196] Multiple techniques of administering a compound exist in the
art including, but not limited to, oral, rectal, topical, aerosol,
injection and parenteral delivery, including intramuscular,
subcutaneous, intravenous, intramedullary injections, intrathecal,
direct intraventricular, intraperitoneal, intranasal and
intraocular injections.
[0197] One may also administer the compound in a local rather than
systemic manner, for example, via injection of the compound
directly into the infected area, often in a depot or sustained
release formulation. Furthermore, one may administer the compound
in a targeted drug delivery system, for example, in a liposome
coated with a tissue-specific antibody. The liposomes will be
targeted to and taken up selectively by the organ.
[0198] The compositions may, if desired, be presented in a pack or
dispenser device which may contain one or more unit dosage forms
containing the active ingredient. The pack may for example comprise
metal or plastic foil, such as a blister pack. The pack or
dispenser device may be accompanied by instructions for
administration. The pack or dispenser may also be accompanied with
a notice associated with the container in form prescribed by a
governmental agency regulating the manufacture, use, or sale of
pharmaceuticals, which notice is reflective of approval by the
agency of the form of the drug for human or veterinary
administration. Such notice, for example, may be the labeling
approved by the U.S. Food and Drug Administration for prescription
drugs, or the approved product insert. Compositions that can
include a compound described herein formulated in a compatible
pharmaceutical carrier may also be prepared, placed in an
appropriate container, and labeled for treatment of an indicated
condition.
Methods of Use:
[0199] Some embodiments described herein relate to a method of
ameliorating, treating and/or preventing a viral infection selected
from a paramyxovirus viral infection and an orthomyxovirus viral
infection, which can include administering to a subject an
effective amount of one or more compounds described herein, or a
pharmaceutical composition that includes one or more compounds
described herein (e.g., a compound of Formula (I), a compound of
Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing). In some
embodiments, the subject is identified as suffering from the viral
infection (for example, a paramyxovirus viral infection or an
orthomyxovirus viral infection).
[0200] Other embodiments described herein relate to a method of
inhibiting viral replication of a virus selected from a
paramyxovirus and an orthomyxovirus, which can include contacting a
cell infected with the virus with an effective amount of a compound
of Formula (I), or a pharmaceutically acceptable salt thereof, an
effective amount of a compound of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of a
compound of Formula (III), or a pharmaceutically acceptable salt
thereof, and/or a pharmaceutical composition that includes one or
more compounds described herein (e.g., a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing).
[0201] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to treat and/or ameliorate a respiratory
syncytial viral (RSV) infection. In some embodiments, an effective
amount of one or more compounds of Formula (I), or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, an
effective amount of one or more compounds of Formula (III), or a
pharmaceutically acceptable salt thereof, and/or a pharmaceutical
composition that includes one or more compounds described herein
(e.g., a compound of Formula (I) a compound of Formula (II) and/or
a compound of Formula (III), or a pharmaceutically acceptable salt
of the foregoing) can be used to prevent a respiratory syncytial
viral infection. In some embodiments, an effective amount of one or
more compounds of Formula (I), or a pharmaceutically acceptable
salt thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to inhibit the replication of a respiratory
syncytial virus. In some embodiments, an effective amount of one or
more compounds of Formula (I), or a pharmaceutically acceptable
salt thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to inhibit the RSV polymerase complex.
[0202] In other embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I) a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to treat and/or ameliorate an influenza
viral infection. In other embodiments, an effective amount of one
or more compounds of Formula (I), or a pharmaceutically acceptable
salt thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to prevent an influenza viral infection. In
some embodiments, an effective amount of one or more compounds of
Formula (I), or a pharmaceutically acceptable salt thereof, one or
more compounds of Formula (II), or a pharmaceutically acceptable
salt thereof, an effective amount of one or more compounds of
Formula (III), or a pharmaceutically acceptable salt thereof,
and/or a pharmaceutical composition that includes one or more
compounds described herein (e.g., a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing) can be used to
inhibit the replication of an influenza virus. In some embodiments,
an effective amount of one or more compounds of Formula (I), or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, an
effective amount of one or more compounds of Formula (III), or a
pharmaceutically acceptable salt thereof, and/or a pharmaceutical
composition that includes one or more compounds described herein
(e.g., a compound of Formula (I), a compound of Formula (II) and/or
a compound of Formula (III), or a pharmaceutically acceptable salt
of the foregoing) can be used to inhibit the influenza polymerase
complex.
[0203] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to treat and/or ameliorate a hendraviral
infection and/or nipahviral infection. In some embodiments, an
effective amount of one or more compounds of Formula (I), or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, an
effective amount of one or more compounds of Formula (III), or a
pharmaceutically acceptable salt thereof, and/or a pharmaceutical
composition that includes one or more compounds described herein
(e.g., a compound of Formula (I) a compound of Formula (II) and/or
a compound of Formula (III), or a pharmaceutically acceptable salt
of the foregoing) can be used to prevent a hendraviral infection
and/or nipahviral infection. In some embodiments, an effective
amount of one or more compounds of Formula (I), or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, an
effective amount of one or more compounds of Formula (III), or a
pharmaceutically acceptable salt thereof, and/or a pharmaceutical
composition that includes one or more compounds described herein
(e.g., a compound of Formula (I), a compound of Formula (II) and/or
a compound of Formula (III), or a pharmaceutically acceptable salt
of the foregoing) can be used to inhibit the replication of a
hendravirus and/or nipahvirus. In some embodiments, an effective
amount of one or more compounds of Formula (I), or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, an
effective amount of one or more compounds of Formula (III), or a
pharmaceutically acceptable salt thereof, and/or a pharmaceutical
composition that includes one or more compounds described herein
(e.g., a compound of Formula (I), a compound of Formula (II) and/or
a compound of Formula (III), or a pharmaceutically acceptable salt
of the foregoing) can be used to inhibit the hendravirus polymerase
complex and/or nipahvirus polymerase complex.
[0204] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to treat and/or ameliorate measles. In some
embodiments, an effective amount of one or more compounds of
Formula (I), or a pharmaceutically acceptable salt thereof, one or
more compounds of Formula (II), or a pharmaceutically acceptable
salt thereof, an effective amount of one or more compounds of
Formula (III), or a pharmaceutically acceptable salt thereof,
and/or a pharmaceutical composition that includes one or more
compounds described herein (e.g., a compound of Formula (I) a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing) can be used to
prevent measles. In some embodiments, an effective amount of one or
more compounds of Formula (I), or a pharmaceutically acceptable
salt thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to inhibit the replication of a measles
virus. In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to inhibit the measles polymerase
complex.
[0205] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to treat and/or ameliorate mumps. In some
embodiments, an effective amount of one or more compounds of
Formula (I), or a pharmaceutically acceptable salt thereof, one or
more compounds of Formula (II), or a pharmaceutically acceptable
salt thereof, an effective amount of one or more compounds of
Formula (III), or a pharmaceutically acceptable salt thereof,
and/or a pharmaceutical composition that includes one or more
compounds described herein (e.g., a compound of Formula (I) a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing) can be used to
prevent mumps. In some embodiments, an effective amount of one or
more compounds of Formula (I), or a pharmaceutically acceptable
salt thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to inhibit the replication of a mumps virus.
In some embodiments, an effective amount of one or more compounds
of Formula (I), or a pharmaceutically acceptable salt thereof, one
or more compounds of Formula (II), or a pharmaceutically acceptable
salt thereof, an effective amount of one or more compounds of
Formula (III), or a pharmaceutically acceptable salt thereof,
and/or a pharmaceutical composition that includes one or more
compounds described herein (e.g., a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing) can be used to
inhibit the mumps polymerase complex.
[0206] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to treat and/or ameliorate a sendai viral
infection. In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I) a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to prevent a sendai viral infection. In some
embodiments, an effective amount of one or more compounds of
Formula (I), or a pharmaceutically acceptable salt thereof, one or
more compounds of Formula (II), or a pharmaceutically acceptable
salt thereof, an effective amount of one or more compounds of
Formula (III), or a pharmaceutically acceptable salt thereof,
and/or a pharmaceutical composition that includes one or more
compounds described herein (e.g., a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing) can be used to
inhibit the replication of a sendai virus. In some embodiments, an
effective amount of one or more compounds of Formula (I), or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, an
effective amount of one or more compounds of Formula (III), or a
pharmaceutically acceptable salt thereof, and/or a pharmaceutical
composition that includes one or more compounds described herein
(e.g., a compound of Formula (I), a compound of Formula (II) and/or
a compound of Formula (III), or a pharmaceutically acceptable salt
of the foregoing) can be used to inhibit the sendai virus
polymerase complex.
[0207] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to treat and/or ameliorate a HPIV-1
infection and/or HPIV-3 infection. In some embodiments, an
effective amount of one or more compounds of Formula (I), or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, an
effective amount of one or more compounds of Formula (III), or a
pharmaceutically acceptable salt thereof, and/or a pharmaceutical
composition that includes one or more compounds described herein
(e.g., a compound of Formula (I) a compound of Formula (II) and/or
a compound of Formula (III), or a pharmaceutically acceptable salt
of the foregoing) can be used to prevent a HPIV-1 infection and/or
HPIV-3 infection. In some embodiments, an effective amount of one
or more compounds of Formula (I), or a pharmaceutically acceptable
salt thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to inhibit the replication of HPIV-1 and/or
HPIV-3. In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to inhibit the HPIV-1 polymerase complex
and/or HPIV-3 polymerase complex.
[0208] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to treat and/or ameliorate a HPIV-2
infection and/or HPIV-4 infection. In some embodiments, an
effective amount of one or more compounds of Formula (I), or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, an
effective amount of one or more compounds of Formula (III), or a
pharmaceutically acceptable salt thereof, and/or a pharmaceutical
composition that includes one or more compounds described herein
(e.g., a compound of Formula (I) a compound of Formula (II) and/or
a compound of Formula (III), or a pharmaceutically acceptable salt
of the foregoing) can be used to prevent a HPIV-2 infection and/or
HPIV-4 infection. In some embodiments, an effective amount of one
or more compounds of Formula (I), or a pharmaceutically acceptable
salt thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to inhibit the replication of HPIV-2 and/or
HPIV-4. In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to inhibit the HPIV-2 polymerase complex
and/or HPIV-4 polymerase complex.
[0209] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to treat and/or ameliorate a human
metapneumoviral infection. In some embodiments, an effective amount
of one or more compounds of Formula (I), or a pharmaceutically
acceptable salt thereof, one or more compounds of Formula (II), or
a pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I) a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to prevent a human metapneumoviral
infection. In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to inhibit the replication of a human
metapneumovirus. In some embodiments, an effective amount of one or
more compounds of Formula (I), or a pharmaceutically acceptable
salt thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to inhibit the human metapneumovirus
polymerase complex.
[0210] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used treat and/or ameliorate an upper respiratory
viral infection caused by a virus selected from a henipavirus, a
morbillivirus, a respirovirus, a rubulavirus, a pneumovirus, a
metapneumovirus and influenza virus. In some embodiments, an
effective amount of one or more compounds of Formula (I), or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, an
effective amount of one or more compounds of Formula (III), or a
pharmaceutically acceptable salt thereof, and/or a pharmaceutical
composition that includes one or more compounds described herein
(e.g., a compound of Formula (I), a compound of Formula (II) and/or
a compound of Formula (III), or a pharmaceutically acceptable salt
of the foregoing) can be used treat and/or ameliorate a lower
respiratory viral infection caused by a virus selected from a
henipavirus, a morbillivirus, a respirovirus, a rubulavirus, a
pneumovirus, a metapneumovirus and influenza virus. In some
embodiments, an effective amount of one or more compounds of
Formula (I), or a pharmaceutically acceptable salt thereof, one or
more compounds of Formula (II), or a pharmaceutically acceptable
salt thereof, an effective amount of one or more compounds of
Formula (III), or a pharmaceutically acceptable salt thereof,
and/or a pharmaceutical composition that includes one or more
compounds described herein (e.g., a compound of Formula (I) a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing) can be used
treat and/or ameliorate one or more symptoms of an infection caused
by a virus selected from a henipavirus, a morbillivirus, a
respirovirus, a rubulavirus, a pneumovirus, a metapneumovirus and
influenza virus (such as those described herein).
[0211] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used treat and/or ameliorate an upper respiratory
viral infection caused by RSV infection, measles, mumps,
parainfluenza infection, metapneumovirus and/or influenza
infection. In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used treat and/or ameliorate a lower respiratory
viral infection caused by RSV infection, measles, mumps,
parainfluenza infection, metapneumovirus and/or influenza
infection. In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I) a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used treat and/or ameliorate one or more symptoms
of an infection caused by RSV infection, measles, mumps,
parainfluenza infection, metapneumovirus and/or influenza infection
(such as those described herein).
[0212] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used treat and/or ameliorate bronchiolitis and/or
tracheobronchitis due to a RSV infection, influenza infection
and/or human parainfluenza virus 3 (HPIV-3) infection. In some
embodiments, an effective amount of one or more compounds of
Formula (I), or a pharmaceutically acceptable salt thereof, one or
more compounds of Formula (II), or a pharmaceutically acceptable
salt thereof, an effective amount of one or more compounds of
Formula (III), or a pharmaceutically acceptable salt thereof,
and/or a pharmaceutical composition that includes one or more
compounds described herein (e.g., a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing) can be used
treat and/or ameliorate pneumonia due to a RSV infection, influenza
infection and/or human parainfluenza virus 3 (HPIV-3) infection. In
some embodiments, an effective amount of one or more compounds of
Formula (I), or a pharmaceutically acceptable salt thereof, one or
more compounds of Formula (II), or a pharmaceutically acceptable
salt thereof, an effective amount of one or more compounds of
Formula (III), or a pharmaceutically acceptable salt thereof,
and/or a pharmaceutical composition that includes one or more
compounds described herein (e.g., a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing) can be used
treat and/or ameliorate croup due to a RSV infection, influenza
infection and/or human parainfluenza virus 1 (HPIV-1)
infection.
[0213] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used treat and/or ameliorate a fever, cough,
runny nose, red eyes, a generalized rash, pneumonia, an ear
infection and/or bronchitis due to measles. In some embodiments, an
effective amount of one or more compounds of Formula (I), or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, an
effective amount of one or more compounds of Formula (III), or a
pharmaceutically acceptable salt thereof, and/or a pharmaceutical
composition that includes one or more compounds described herein
(e.g., a compound of Formula (I), a compound of Formula (II) and/or
a compound of Formula (III), or a pharmaceutically acceptable salt
of the foregoing) can be used treat and/or ameliorate swelling of
the salivary glands, fever, loss of appetite and/or fatigue due to
mumps.
[0214] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to prevent an influenza viral infection. In
some embodiments, the influenza viral infection can be an influenza
A viral infection. In other embodiments, the influenza viral
infection can be an influenza B viral infection. In still other
embodiments, the influenza viral infection can be an influenza C
viral infection. In some embodiments, one or more compounds of
Formula (I), or a pharmaceutically acceptable salt thereof, one or
more compounds of Formula (II), or a pharmaceutically acceptable
salt thereof, and/or one or more compounds of Formula (III), or a
pharmaceutically acceptable salt thereof, can be used to treat
and/or ameliorate one or more subtypes of influenza. For example,
one or more compounds of Formula (I), or a pharmaceutically
acceptable salt thereof, one or more compounds of Formula (II), or
a pharmaceutically acceptable salt thereof, and/or one or more
compounds of Formula (III), or a pharmaceutically acceptable salt
thereof, can be used to treat H1N1 and/or H3N2.
[0215] In some embodiments, an effective amount of one or more
compounds of Formula (I), or a pharmaceutically acceptable salt
thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, an effective amount of
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, and/or a pharmaceutical composition that
includes one or more compounds described herein (e.g., a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing) can be used to prevent a human parainfluenza viral
infection. In some embodiments, the human parainfluenza viral
infection can be a human parainfluenza virus 1 (HPIV-1). In other
embodiments, the human parainfluenza viral infection can be a human
parainfluenza virus 2 (HPIV-2). In other embodiments, the human
parainfluenza viral infection can be a human parainfluenza virus 3
(HPIV-3). In other embodiments, the human parainfluenza viral
infection can be a human parainfluenza virus 4 (HPIV-4). In some
embodiments, one or more compounds of Formula (I), or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, and/or
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, can be used to treat and/or ameliorate one
or more subtypes of human parainfluenza virus. For example, one or
more compounds of Formula (I), or a pharmaceutically acceptable
salt thereof, one or more compounds of Formula (II), or a
pharmaceutically acceptable salt thereof, and/or one or more
compounds of Formula (III), or a pharmaceutically acceptable salt
thereof, can be used to treat HPIV-1 and/or HPIV-3.
[0216] The one or more compounds of Formula (I) or a
pharmaceutically acceptable salt thereof, one or more compounds of
Formula (II), or a pharmaceutically acceptable salt thereof, and/or
one or more compounds of Formula (III), or a pharmaceutically
acceptable salt thereof, that can be used to treat, ameliorate
and/or prevent a paramyxovirus and/or or an orthomyxovirus viral
infection can be a compound of Formula (I), or pharmaceutically
acceptable salt thereof, and/or a compound of Formula (II), or a
pharmaceutically acceptable salt thereof, and/or a compound of
Formula (III), or a pharmaceutically acceptable salt thereof,
provided in any of the embodiments described in paragraphs
[0085]-[0171].
[0217] As used herein, the terms "prevent" and "preventing," mean a
subject does not develop an infection because the subject has an
immunity against the infection, or if a subject becomes infected,
the severity of the disease is less compared to the severity of the
disease if the subject has not been administered/received the
compound. Examples of forms of prevention include prophylactic
administration to a subject who has been or may be exposed to an
infectious agent, such as a paramyxovirus (e.g., RSV) and/or an
orthomyxovirus (e.g., influenza).
[0218] As used herein, the terms "treat," "treating," "treatment,"
"therapeutic," and "therapy" do not necessarily mean total cure or
abolition of the disease or condition. Any alleviation of any
undesired signs or symptoms of a disease or condition, to any
extent can be considered treatment and/or therapy. Furthermore,
treatment may include acts that may worsen the subject's overall
feeling of well-being or appearance.
[0219] The terms "therapeutically effective amount" and "effective
amount" are used to indicate an amount of an active compound, or
pharmaceutical agent, that elicits the biological or medicinal
response indicated. For example, a therapeutically effective amount
of compound can be the amount needed to prevent, alleviate or
ameliorate symptoms of disease or prolong the survival of the
subject being treated This response may occur in a tissue, system,
animal or human and includes alleviation of the signs or symptoms
of the disease being treated. Determination of an effective amount
is well within the capability of those skilled in the art, in view
of the disclosure provided herein. The therapeutically effective
amount of the compounds disclosed herein required as a dose will
depend on the route of administration, the type of animal,
including human, being treated, and the physical characteristics of
the specific animal under consideration. The dose can be tailored
to achieve a desired effect, but will depend on such factors as
weight, diet, concurrent medication and other factors which those
skilled in the medical arts will recognize.
[0220] Various indicators for determining the effectiveness of a
method for treating a viral infection, such as a paramyxovirus
and/or an orthomyxovirus infection, are known to those skilled in
the art. Example of suitable indicators include, but are not
limited to, a reduction in viral load, a reduction in viral
replication, a reduction in time to seroconversion (virus
undetectable in patient serum), a reduction of morbidity or
mortality in clinical outcomes, and/or other indicator of disease
response.
[0221] In some embodiments, an effective amount of a compound of
Formulae (I), (II) and/or (III), or a pharmaceutically acceptable
salt of the foregoing, is an amount that is effective to reduce
viral titers to undetectable levels, for example, to about 1000 to
about 5000, to about 500 to about 1000, or to about 100 to about
500 genome copies/mL serum. In some embodiments, an effective
amount of a compound of Formulae (I), (II) and/or (III), or a
pharmaceutically acceptable salt of the foregoing, is an amount
that is effective to reduce viral load compared to the viral load
before administration of the compound of Formulae (I), (II) and/or
(III), or a pharmaceutically acceptable salt of the foregoing. For
example, wherein the viral load is measure before administration of
the compound of Formulae (I), (II) and/or (III), or a
pharmaceutically acceptable salt of the foregoing, and again after
completion of the treatment regime with the compound of Formulae
(I), (II) and/or (III), or a pharmaceutically acceptable salt of
the foregoing (for example, 1 week after completion). In some
embodiments, an effective amount of a compound of Formulae (I),
(II) and/or (III), or a pharmaceutically acceptable salt of the
foregoing, can be an amount that is effective to reduce viral load
to lower than about 100 genome copies/mL serum. In some
embodiments, an effective amount of a compound of Formulae (I),
(II) and/or (III), or a pharmaceutically acceptable salt of the
foregoing, is an amount that is effective to achieve a reduction in
viral titer in the serum of the subject in the range of about
1.5-log to about a 2.5-log reduction, about a 3-log to about a
4-log reduction, or a greater than about 5-log reduction compared
to the viral load before administration of the compound of Formulae
(I), (II) and/or (III), or a pharmaceutically acceptable salt of
the foregoing. For example, wherein the viral load is measure
before administration of the compound of Formulae (I), (II) and/or
(III), or a pharmaceutically acceptable salt of the foregoing, and
again after completion of the treatment regime with the compound of
Formulae (I), (II) and/or (III), or a pharmaceutically acceptable
salt of the foregoing (for example, 1 week after completion).
[0222] In some embodiments, a compound of Formulae (I), (II) and/or
(III), or a pharmaceutically acceptable salt of the foregoing, can
result in at least a 1, 2, 3, 4, 5, 10, 15, 20, 25, 50, 75,
100-fold or more reduction in the replication of a paramyxovirus
and/or an orthomyxovirus relative to pre-treatment levels in a
subject, as determined after completion of the treatment regime
(for example, 1 week after completion). In some embodiments, a
compound of Formulae (I), (II) and/or (III), or a pharmaceutically
acceptable salt of the foregoing, can result in a reduction of the
replication of a paramyxovirus and/or an orthomyxovirus relative to
pre-treatment levels in the range of about 2 to about 5 fold, about
10 to about 20 fold, about 15 to about 40 fold, or about 50 to
about 100 fold. In some embodiments, a compound of Formulae (I),
(II) and/or (III), or a pharmaceutically acceptable salt of the
foregoing, can result in a reduction of paramyxovirus replication
in the range of 1 to 1.5 log, 1.5 log to 2 log, 2 log to 2.5 log,
2.5 to 3 log, 3 log to 3.5 log or 3.5 to 4 log more reduction of
paramyxovirus replication compared to the reduction of
paramyxovirus reduction achieved by ribavirin (Virazole.RTM.), or
may achieve the same reduction as that of ribavirin (Virazole.RTM.)
therapy in a shorter period of time, for example, in one week, two
weeks, one month, two months, or three months, as compared to the
reduction achieved after six months of ribavirin (Virazole.RTM.)
therapy. In some embodiments, a compound of Formulae (I), (II)
and/or (III), or a pharmaceutically acceptable salt of the
foregoing, can result in a reduction of orthomyxovirus replication
in the range of 1 to 1.5 log, 1.5 log to 2 log, 2 log to 2.5 log,
2.5 to 3 log, 3 log to 3.5 log or 3.5 to 4 log more reduction of
orthomyxovirus replication compared to the reduction of
orthomyxovirus reduction achieved by oseltamivir (Tamiflu.RTM.), or
may achieve the same reduction as that of oseltamivir
(Tamiflu.RTM.) therapy in a shorter period of time, for example, in
one week, two weeks, one month, two months, or three months, as
compared to the reduction achieved after six months of oseltamivir
(Tamiflu.RTM.) therapy.
[0223] In some embodiments, an effective amount of a compound of
Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing, is an amount that is effective to achieve a sustained
viral response, for example, non-detectable or substantially
non-detectable paramyxovirus and/or orthomyxovirus RNA (e.g., less
than about 500, less than about 400, less than about 200, or less
than about 100 genome copies per milliliter serum) is found in the
subject's serum for a period of at least about one week, two weeks,
one month, at least about two months, at least about three months,
at least about four months, at least about five months, or at least
about six months following cessation of therapy.
[0224] After a period of time, infectious agents can develop
resistance to one or more therapeutic agents. The term "resistance"
as used herein refers to a viral strain displaying a delayed,
lessened and/or null response to a therapeutic agent(s). For
example, after treatment with an antiviral agent, the viral load of
a subject infected with a resistant virus may be reduced to a
lesser degree compared to the amount in viral load reduction
exhibited by a subject infected with a non-resistant strain. In
some embodiments, a compound of Formula (I), a compound of Formula
(II) and/or a compound of Formula (III), or a pharmaceutically
acceptable salt of the foregoing, can be administered to a subject
infected with RSV that is resistant to one or more different
anti-RSV agents (for example, ribavirin). In some embodiments,
development of resistant RSV strains can be delayed when subjects
are treated with a compound of Formula (I), a compound of Formula
(II) and/or a compound of Formula (III), or a pharmaceutically
acceptable salt of the foregoing, compared to the development of
RSV strains resistant to other RSV drugs. In some embodiments, a
compound of Formula (I), a compound of Formula (II) and/or a
compound of Formula (III), or a pharmaceutically acceptable salt of
the foregoing, can be administered to a subject infected with an
influenza virus that is resistant to one or more different
anti-influenza agents (for example, amantadine and rimantadine). In
some embodiments, development of resistant influenza strains can be
delayed when subjects are treated with a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing, compared to the
development of influenza strains resistant to other influenza
drugs.
[0225] In some embodiments, a compound of Formula (I), a compound
of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing, can decrease the
percentage of subjects that experience complications from a RSV
viral infection compared to the percentage of subjects that
experience complication being treated with ribavirin. In some
embodiments, a compound of Formula (I), a compound of Formula (II)
and/or a compound of Formula (III), or a pharmaceutically
acceptable salt of the foregoing, can decrease the percentage of
subjects that experience complications from an influenza viral
infection compared to the percentage of subjects that experience
complication being treated with oseltamivir. For example, the
percentage of subjects being treated with a compound of Formula
(I), a compound of Formula (II) and/or a compound of Formula (III),
or a pharmaceutically acceptable salt of the foregoing, that
experience complications can be 10%, 25%, 40%, 50%, 60%, 70%, 80%
and 90% less compared to subjects being treated with ribavirin or
oseltamivir.
[0226] In some embodiments, a compound of Formula (I), a compound
of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing, or a
pharmaceutical composition that includes a compound described
herein, can be used in combination with one or more additional
agent(s). In some embodiments, a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing, can be used in
combination with one or more agents currently used for treating
RSV. For example, the additional agent can be ribavirin,
palivizumab and RSV-IGIV. For the treatment of RSV, additional
agents include but are not limited to ALN-RSV01 (Alnylam
Pharmaceuticals), BMS-433771
(1-cyclopropyl-3-[[1-(4-hydroxybutyl)benzimidazol-2-yl]methyl]imidazo[4,5-
-c]pyridin-2-one), RFI-641
((4,4''-bis-{4,6-bis-[3-(bis-carbamoylmethyl-sulfamoyl)-phenylamino]-(1,3-
,5)triazin-2-ylamino}-biphenyl-2,2''-disulfonic-acid)), RSV604
((S)-1-(2-fluorophenyl)-3-(2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]di--
azepin-3-yl)-urea), MDT-637
((4Z)-2-methylsulfanyl-4-[(E)-3-thiophen-2-ylprop-2-enylidene]-1,3-thiazo-
l-5-one), BTA9881, TMC-353121 (Tibotec), MBX-300, YM-53403
(N-cyclopropyl-6-[4-[(2-phenylbenzoyl)amino]benzoyl]-4,5-dihydrothieno[3,-
2-d][1]benzazepine-2-carboxamide), motavizumab (Medi-524,
MedImmune), Medi-559, Medi-534, Medi-557, RV568 and a RSV-F
Particle Vaccine (Novavax). In some embodiments, a compound of
Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing, can be used in combination with one or more agents
currently used for treating influenza. For example, the additional
agent can be amantadine, rimantadine, zanamivir and oseltamivir.
For the treatment of influenza, additional agents include but are
not limited to peramivir
((1S,2S,3S,4R)-3-[(1S)-1-acetamido-2-ethylbutyl]-4-(diaminomethylideneami-
no)-2-hydroxycyclopentane-1-carboxylic acid), laninamivir
((4S,5R,6R)-5-acetamido-4-carbamimidamido-6-[(1R,2R)-3-hydroxy-2-methoxyp-
ropyl]-5,6-dihydro-4H-pyran-2-carboxylic acid), favipiravir (T-705,
6-fluoro-3-hydroxy-2-pyrazinecarboxamide), fludase (DAS181,
NexBio), ADS-8902 (Adamas Pharmaceuticals), IFN-b (Synairgen),
beraprost
(4-[2-hydroxy-1-[(E)-3-hydroxy-4-methyloct-1-en-6-ynyl]-2,3,3a,8b-tetrahy-
dro-1H-cyclopenta[b][1]benzofuran-5-yl]butanoic acid), Neugene.RTM.
and VGX-3400X (Inovio).
[0227] In some embodiments, a compound of Formula (I), a compound
of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing, can be
administered with one or more additional agent(s) together in a
single pharmaceutical composition. In some embodiments, a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing, can be administered with one or more additional agent(s)
as two or more separate pharmaceutical compositions. For example, a
compound of Formula (I), a compound of Formula (II) and/or a
compound of Formula (III), or a pharmaceutically acceptable salt of
the foregoing, can be administered in one pharmaceutical
composition, and at least one of the additional agents can be
administered in a second pharmaceutical composition. If there are
at least two additional agents, one or more of the additional
agents can be in a first pharmaceutical composition that includes a
compound of Formula (I), a compound of Formula (II) and/or a
compound of Formula (III), or a pharmaceutically acceptable salt of
the foregoing, and at least one of the other additional agent(s)
can be in a second pharmaceutical composition.
[0228] The order of administration of a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing, with one or more
additional agent(s) can vary. In some embodiments, a compound of
Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing, can be administered prior to all additional agents. In
other embodiments, a compound of Formula (I), a compound of Formula
(II) and/or a compound of Formula (III), or a pharmaceutically
acceptable salt of the foregoing, can be administered prior to at
least one additional agent. In still other embodiments, a compound
of Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing, can be administered concomitantly with one or more
additional agent(s). In yet still other embodiments, a compound of
Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing, can be administered subsequent to the administration of
at least one additional agent. In some embodiments, a compound of
Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt of the
foregoing, can be administered subsequent to the administration of
all additional agents.
[0229] A potential advantage of utilizing a compound of Formula
(I), a compound of Formula (II) and/or a compound of Formula (III),
or a pharmaceutically acceptable salt of the foregoing, in
combination with one or more additional agent(s) described in
paragraph [0222], including pharmaceutically acceptable salts and
prodrugs thereof, may be a reduction in the required amount(s) of
one or more compounds of paragraph [0222] (including
pharmaceutically acceptable salts and prodrugs thereof) that is
effective in treating a disease condition disclosed herein (for
example, RSV and/or influenza), as compared to the amount required
to achieve same therapeutic result when one or more compounds
described in paragraph [0222], including pharmaceutically
acceptable salts and prodrugs thereof, are administered without a
compound of Formula (I), a compound of Formula (II) and/or a
compound of Formula (III), or a pharmaceutically acceptable salt
the foregoing. For example, the amount of a compound described in
paragraph [0222], including a pharmaceutically acceptable salt and
prodrug thereof, can be less compared to the amount of the compound
described in paragraph [0222], including a pharmaceutically
acceptable salt and prodrug thereof, needed to achieve the same
viral load reduction when administered as a monotherapy. Another
potential advantage of utilizing a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt of the foregoing, in combination
with one or more additional agent(s) described in paragraph [0222],
including pharmaceutically acceptable salts and prodrugs thereof,
is that the use of two or more compounds having different mechanism
of actions can create a higher barrier to the development of
resistant viral strains compared to the barrier when a compound is
administered as monotherapy.
[0230] Additional advantages of utilizing a compound of Formula
(I), a compound of Formula (II) and/or a compound of Formula (III),
or a pharmaceutically acceptable salt the foregoing, in combination
with one or more additional agent(s) described in paragraph [0222],
including pharmaceutically acceptable salts and prodrugs thereof,
may include little to no cross resistance between a compound of
Formula (I), a compound of Formula (II) and/or a compound of
Formula (III), or a pharmaceutically acceptable salt the foregoing,
and one or more additional agent(s) described in paragraph [0222]
(including pharmaceutically acceptable salts and prodrugs thereof);
different routes for elimination of a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt the foregoing, and one or more
additional agent(s) described in paragraph [0222] (including
pharmaceutically acceptable salts and prodrugs thereof); little to
no overlapping toxicities between a compound of Formula (I), a
compound of Formula (II) and/or a compound of Formula (III), or a
pharmaceutically acceptable salt the foregoing, and one or more
additional agent(s) described in paragraph [0222] (including
pharmaceutically acceptable salts and prodrugs thereof); little to
no significant effects on cytochrome P450; and/or little to no
pharmacokinetic interactions between a compound of Formula (I), or
a pharmaceutically acceptable salt thereof, and one or more
additional agent(s) described in paragraph [0222] (including
pharmaceutically acceptable salts and prodrugs thereof).
[0231] As will be readily apparent to one skilled in the art, the
useful in vivo dosage to be administered and the particular mode of
administration will vary depending upon the age, weight, the
severity of the affliction, and mammalian species treated, the
particular compounds employed, and the specific use for which these
compounds are employed. The determination of effective dosage
levels, that is the dosage levels necessary to achieve the desired
result, can be accomplished by one skilled in the art using routine
methods, for example, human clinical trials and in vitro
studies.
[0232] The dosage may range broadly, depending upon the desired
effects and the therapeutic indication. Alternatively dosages may
be based and calculated upon the surface area of the patient, as
understood by those of skill in the art. Although the exact dosage
will be determined on a drug-by-drug basis, in most cases, some
generalizations regarding the dosage can be made. The daily dosage
regimen for an adult human patient may be, for example, an oral
dose of between 0.01 mg and 3000 mg of each active ingredient,
preferably between 1 mg and 700 mg, e.g. 5 to 200 mg. The dosage
may be a single one or a series of two or more given in the course
of one or more days, as is needed by the subject. In some
embodiments, the compounds will be administered for a period of
continuous therapy, for example for a week or more, or for months
or years.
[0233] In instances where human dosages for compounds have been
established for at least some condition, those same dosages may be
used, or dosages that are between about 0.1% and 500%, more
preferably between about 25% and 250% of the established human
dosage. Where no human dosage is established, as will be the case
for newly-discovered pharmaceutical compositions, a suitable human
dosage can be inferred from ED.sub.50 or ID.sub.50 values, or other
appropriate values derived from in vitro or in vivo studies, as
qualified by toxicity studies and efficacy studies in animals.
[0234] In cases of administration of a pharmaceutically acceptable
salt, dosages may be calculated as the free base. As will be
understood by those of skill in the art, in certain situations it
may be necessary to administer the compounds disclosed herein in
amounts that exceed, or even far exceed, the above-stated,
preferred dosage range in order to effectively and aggressively
treat particularly aggressive diseases or infections.
[0235] Dosage amount and interval may be adjusted individually to
provide plasma levels of the active moiety which are sufficient to
maintain the modulating effects, or minimal effective concentration
(MEC). The MEC will vary for each compound but can be estimated
from in vitro data. Dosages necessary to achieve the MEC will
depend on individual characteristics and route of administration.
However, HPLC assays or bioassays can be used to determine plasma
concentrations. Dosage intervals can also be determined using MEC
value. Compositions should be administered using a regimen which
maintains plasma levels above the MEC for 10-90% of the time,
preferably between 30-90% and most preferably between 50-90%. In
cases of local administration or selective uptake, the effective
local concentration of the drug may not be related to plasma
concentration.
[0236] It should be noted that the attending physician would know
how to and when to terminate, interrupt, or adjust administration
due to toxicity or organ dysfunctions. Conversely, the attending
physician would also know to adjust treatment to higher levels if
the clinical response were not adequate (precluding toxicity). The
magnitude of an administrated dose in the management of the
disorder of interest will vary with the severity of the condition
to be treated and to the route of administration. The severity of
the condition may, for example, be evaluated, in part, by standard
prognostic evaluation methods. Further, the dose and perhaps dose
frequency, will also vary according to the age, body weight, and
response of the individual patient. A program comparable to that
discussed above may be used in veterinary medicine.
[0237] Compounds disclosed herein can be evaluated for efficacy and
toxicity using known methods. For example, the toxicology of a
particular compound, or of a subset of the compounds, sharing
certain chemical moieties, may be established by determining in
vitro toxicity towards a cell line, such as a mammalian, and
preferably human, cell line. The results of such studies are often
predictive of toxicity in animals, such as mammals, or more
specifically, humans. Alternatively, the toxicity of particular
compounds in an animal model, such as mice, rats, rabbits, or
monkeys, may be determined using known methods. The efficacy of a
particular compound may be established using several recognized
methods, such as in vitro methods, animal models, or human clinical
trials. When selecting a model to determine efficacy, the skilled
artisan can be guided by the state of the art to choose an
appropriate model, dose, route of administration and/or regime.
EXAMPLES
[0238] Additional embodiments are disclosed in further detail in
the following examples, which are not in any way intended to limit
the scope of the claims.
Example 1
Preparation of Compound (1a)
##STR00161##
[0240] Preparation of (P1-2): To an ice cooled solution of P1-1
(10.0 g, 40.8 mmol) in dry pyridine (100 mL) was added TBSCl in
pyridine (1M, 53 mL) dropwise at room temperature (R.T.). The
reaction mixture was stirred at R.T. for 16 hours. The reaction
mixture was then quenched with water, concentrated to give a
residue. The residue was separated by ethyl acetate (EA) and
saturated NaHCO.sub.3 aq. solution. The organic phase was dried and
concentrated. The residue was purified on a silica gel column (5%
MeOH in DCM) to give a crude 5'-O-TBS protected intermediate as a
white solid (13.4 g, 91%). The intermediate was dissolved in
anhydrous DCM (100 mL) and sym-collidine (17.9 g, 149.2 mmol),
AgNO.sub.3 (25 g, 149.2 mmol) and MMTrCl (45 g, 149.2 mmol) were
added. The mixture was stirred at R.T. for 16 hours. The mixture
was quenched with water, and the organic layer was separated and
concentrated. The residue purified on a silica gel column (30% PE
in EA) to give the crude product. The crude product was dissolved
in 1M TBAF (50 mL) in THF. The mixture was stirred at R.T. for 2
hours. The solvent was removed, and the residue was purified on a
silica gel column (50% PE in EA) to give P1-2 as a white solid
(21.4 g, 66% for three steps).
[0241] Preparation of (P1-3): To a solution of pyridine (521 mg,
6.59 mmol) in anhydrous DMSO (5 mL) was added TFA (636 mg, 5.58
mmol) dropwise at 10.degree. C. under nitrogen. The reaction
mixture was stirred until the solution became clear. The solution
was then added into a mixture of P1-2 (4.0 g, 5.07 mmol) and DCC
(3.86 g, 18.76 mmol) in anhydrous DMSO (18 mL) at R.T. under
nitrogen. The reaction mixture was stirred at 30.degree. C.
overnight. Water (80 mL) was added into the mixture, diluted with
EtOAc (100 mL) and filtered. The filtrate was extracted with DCM
(100 mL.times.6). The organic layer was washed with saturated aq.
NaHCO.sub.3, dried over Na.sub.2SO.sub.4 and concentrated in vacuo.
The residue was purified on a silica gel column eluted with 1% MeOH
in DCM to give the intermediate (3.5 g, 87.7%) as a yellow solid.
The intermediate (3.5 g, 4.45 mmol) was dissolved in dioxane (25
mL) and aq. HCHO (668 mg, 22.25 mmol) was added at R.T. 2N NaOH
(4.5 mL, 8.9 mmol) was then added. The reaction mixture was stirred
at 30.degree. C. overnight. NaBH.sub.4 (593 mg, 15.6 mmol) was
added in by portions at 5.degree. C., and the mixture was stirred
at R.T. for 15 min. The reaction was quenched with water, and the
mixture was extracted with EtOAc (100 mL.times.3). The organic
layer was dried over Na.sub.2SO.sub.4 and concentrated in vacuo.
The residue was purified on a silica gel column eluted with 1% MeOH
in DCM to give P1-3 as a yellow solid (2.5 g, 67%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 6.82-7.50 (m, 29H), 5.40 (d, J=23.2
Hz, 1H), 4.99 (d, J=7.6 Hz, 1H), 4.46 (dd, J.sub.1=6.0 Hz,
J.sub.2=54.4 Hz, 1H), 3.94 (dd, J.sub.1=4.4 Hz, J.sub.2=12.4 Hz,
1H), 3.78 (s, 6H), 3.42-3.69 (m, 2H), 2.71-3.05 (m, 2H), 2.45 (m,
1H).
[0242] Preparation of (P1-4): To an ice cooled solution of P1-3
(4.0 g, 4.9 mmol) in dry pyridine (20 mL) was added dropwise TBSCl
in pyridine (1M, 5.88 mL). The reaction mixture was stirred at R.T.
for 16 hours. The reaction mixture was then quenched with water,
concentrated to give a residue. The residue was separated in EA and
saturated aq. NaHCO.sub.3. The organic layer was separated and
dried, and then concentrated. The residue was purified on a silica
gel column (1% MeOH in DCM) to give the intermediate as a yellow
solid (3.2 g, 70%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
7.53-6.83 (m, 29H), 5.51 (d, J=21.2 Hz, 1H), 4.98 (d, J=7.6 Hz,
1H), 4.67 (dd, J.sub.1=5.6 Hz, J.sub.2=22.4 Hz, 1H), 4.22 (dd,
J.sub.1=5.6 Hz, J.sub.2=53.2 Hz, 1H), 4.07 (m, 1H), 3.89 (m, 1H),
3.80 (s, 6H), 3.70-3.67 (m, 1H), 3.03-2.98 (m, 1H), 2.26 (m, 1H),
0.93 (s, 9H), 0.10 (s, 6H).
[0243] The obtained intermediate was dissolved in anhydrous DCM (20
mL) and collidine (360 mg, 3 mmol), and AgNO.sub.3 (500 mg, 3 mmol)
and MMTrCl (606 mg, 2 mmol) were added. The mixture was stirred at
R.T. for 16 hours. The reaction mixture was quenched with water,
and the organic layer was separated and concentrated. The residue
was purified on a silica gel column (0.5% MeOH in DCM) to give the
fully protected intermediate as a yellow solid (3.3 g, 80%). The
intermediate was dissolved in 1M TBAF in THF (5 mL) and was stirred
at R.T. for 2 hours. The solution was concentrated, and the residue
was purified on a silica gel column (1% MeOH in DCM) to give a
mixture of P1-3 and P1-4, which was separated by HPLC separation
(MeCN and 0.1% HCOOH in water) to give P1-4 as a white solid (1.5
g, 25%).
[0244] Preparation of (P1-5): P1-4 (1.5 g, 1.22 mmol) was suspended
in anhydrous DCM (50 mL), and Dess Martin periodinane (1.2 g, 2.73
mmol) was added at 0.degree. C. The reaction mixture was stirred at
R.T. for 3 hours. The reaction mixture was then quenched with
saturated aq. Na.sub.2S.sub.2O.sub.3 and Na.sub.2CO.sub.3. The
organic layer was separated and dried, and then concentrated to
give the aldehyde intermediate as a white solid.
[0245] A solution of ClCH.sub.2PPh.sub.3Br (2.19 g, 5.6 mmol) in
anhydrous THF (40 mL) was cooled to -78.degree. C. n-BuLi (2.5 M,
2.3 mL) was added in dropwise. After the addition, the mixture was
stirred at 0.degree. .degree. C. for 2 hours. A solution of the
aldehyde in anhydrous THF (10 mL) was then added. The mixture was
stirred at R.T. for 16 hours. The reaction was quenched with
saturated NH.sub.4Cl aq. and extracted by EA. The organic layer was
separated, dried and concentrated. The residue was purified on a
silica gel column (1% MeOH in DCM) to give the intermediate as a
yellow solid (1.1 g, 73%). To a solution of the intermediate (1.1
g, 0.98 mmol) in anhydrous THF (40 mL) was added n-BuLi (2.5M, 6
mL) -78.degree. C. dropwise. The mixture was stirred at -78.degree.
C. for 5 hours and then quenched with a saturated NH.sub.4Cl aq.
solution. The mixture was extracted with EA. The organic layer was
separated, dried and concentrated. The residue was purified on a
silica gel column (2% MeOH in DCM) to give P1-5 as a yellow solid
(910 mg, 86%).
[0246] Preparation of (1a): P1-5 (910 mg, 0.84 mmol) was suspended
in 80% CH.sub.3COOH (50 mL), and the reaction mixture was stirred
at 40.degree. C. for 15 hours. The solvents were evaporated, and
the residue was co-evaporated with toluene to remove traces of acid
and water. The residue was purified by HPLC separation (MeCN and
0.1% HCOOH in water) to give pure compound 1a as a white solid (101
mg, 45%). .sup.1H NMR (MeOD, 400 MHz) .delta. 7.90 (d, J=7.2 Hz,
1H), 6.04 (d, J=19.6 Hz, 1H), 5.87 (d, J=7.6 Hz, 1H), 5.00 (dd,
J.sub.1=5.2 Hz, J.sub.2=53.6 Hz, 1H), 4.47 (dd, J.sub.1=5.2 Hz,
J.sub.2=22.8 Hz, 1H), 3.86 (d, J=12.4 Hz, 1H), 3.73 (d, J=12.4 Hz,
1H), 3.08 (s, 1H); ESI-TOF-MS: m/z 270.09 [M+H].sup.+, 539.17
[2M+H].sup.+.
Example 2
Preparation of Compound (2a)
##STR00162##
[0248] To a stirred solution of compound 1a (50 mg, 0.186 mmol) in
anhydrous THF (3 mL) was added dropwise a solution of t-BuMgCl
(0.37 mL, 1M in THF) at -78.degree. C. The mixture was then stirred
at 0.degree. C. for 30 min and re-cooled to -78.degree. C. A
solution of phenyl (isopropoxy-L-alaninyl) phosphorochloridate (104
mg, 0.4 mmol) in THF (0.5 mL) was added dropwise. After addition,
the mixture was stirred at 25.degree. C. for 16 hours. The reaction
was quenched with HCOOH (80% aq.) at 0.degree. C. The solvent was
removed, and the residue was purified on silica gel (DCM:MeOH=50:1
to 10:1) to give compound 2a as a white solid (a mixture of two P
isomers, 8.0 mg, 7.9%). .sup.1H NMR (MeOD, 400 MHz) .delta. 7.71,
7.68 (2d, J=7.6 Hz, 1H), 7.17-7.37 (m, 5H), 6.02, 6.00 (2d, J=20.4
Hz, 1H), 5.90, 5.86 (2d, J=7.6 Hz, 1H), 5.03-5.18 (m, 1H),
4.91-4.99 (m, 1H), 4.45-4.55 (m, 1H), 4.34-4.43 (m, 1H), 4.26-4.33
(m, 1H), 3.87-3.95 (m, 1H), 3.25, 3.22 (2s, 1H), 1.29-1.34 (m, 3H),
1.20-1.22 (m, 6H). .sup.31P NMR (MeOD, 162 MHz) .delta. 3.44, 3.27.
ESI-LCMS: m/z 539.0 [M+H].sup.+.
Example 3
Preparation of Compound (3a)
##STR00163## ##STR00164##
[0250] Preparation of (P3-2): To a solution of P3-1 (100.0 g, 406.5
mmol) in pyridine (750 mL) was added DMTrCl (164.9 g, 487.8 mmol).
The solution was stirred at R.T. for 15 hours. MeOH (300 mL) was
added, and the mixture was concentrated to dryness under reduced
pressure. The residue was dissolved in EtOAc and washed with water.
The organic layer was dried over Na.sub.2SO.sub.4 and concentrated.
The residue was dissolved in DCM (500 mL). Imidazole (44.3 g, 650.4
mmol) and TBSCl (91.9 g, 609.8 mmol) was added. The reaction
mixture was stirred at R.T. for 14 hours. The reaction solution was
washed with NaHCO.sub.3 and brine. The organic layer was dried over
Na.sub.2SO.sub.4, and concentrated to give the crude as a light
yellow solid. The crude (236.4 g, 356.6 mmol) was dissolved in 80%
HOAc aq. solution (500 mL). The mixture was stirred at R.T. for 15
hours. The mixture was diluted with EtOAc and washed with a
NaHCO.sub.3 solution and brine. The organic layer was dried over
Na.sub.2SO.sub.4 and purified by silica gel column chromatography
(1-2% MeOH in DCM) to give P3-2 (131.2 g, 89.6%) as a light yellow
solid. .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.39 (s, 1H), 7.88
(d, J=7.2 Hz, 1H), 5.89 (dd, J=18.0 Hz, J.sub.2=2.0 Hz, 1H), 5.64
(d, J=8.0 Hz, 1H), 5.21 (dd, J.sub.1=J.sub.2=7.2 Hz, 1H),
5.18.about.5.03 (m, 1H), 4.37.about.4.29 (m, 1H), 3.86 (dd,
J.sub.1=J.sub.2=3.2 Hz, 3H), 3.78.about.3.73 (m, 1H),
3.51.about.3.56 (m, 1H), 3.31 (s, 1H), 0.89 (s, 9H), 0.11 (s, 6H);
ESI-MS: m/z 802 [M+H].sup.+.
[0251] Preparation of (P3-3): To a solution of P3-2 (131.2 g, 364.0
mmol) in anhydrous CH.sub.3CN (1200 mL) was added IBX (121.2 g,
432.8 mmol) at R.T. The reaction mixture was refluxed for 3 hours
and then cooled to 0.degree. C. The precipitate was filtered-off,
and the filtrate was concentrated to give the crude aldehyde (121.3
g) as a yellow solid. The aldehyde was dissolved in 1,4-dioxane
(1000 mL). 37% CH.sub.2O (81.1 mL, 1.3536 mol) and 2M NaOH aq.
solution (253.8 mL, 507.6 mmol) were added. The mixture was stirred
at R.T. for 2 hours and then neutralized with AcOH to pH=7. To the
solution were added EtOH (400 mL) and NaBH.sub.4 (51.2 g, 1.354
mol). The mixture was stirred at R.T. for 30 minutes. The mixture
was quenched with saturated aq. NH.sub.4Cl and extracted with EA.
The organic layer was dried over Na.sub.2SO.sub.4 and concentrated.
The residue was purified by silica gel column chromatography (1-3%
MeOH in DCM) to give P3-3 (51.4 g, 38.9%) as a white solid.
[0252] Preparation of (P3-4): To a solution of P3-3 (51.4 g, 131.6
mmol) in anhydrous DCM (400 mL) were added pyridine (80 mL) and
DMTrCl (49.1 g, 144.7 mmol) at 0.degree. C. The reaction was
stirred at R.T. for 14 hours, and then treated with MeOH (30 mL).
The solvent was removed, and the residue was purified by silica gel
column chromatography (1-3% MeOH in DCM) to give a mono-DMTr
protected intermediate as a yellow foam (57.4 g, 62.9%). To the
intermediate (57.4 g, 82.8 mmol) in CH.sub.2Cl.sub.2 (400 mL) was
added imidazole (8.4 g, 124.2 mmol) and TBDPSCl (34.1 g, 124.2
mmol). The mixture was stirred at R.T. for 14 hours. The
precipitate was filtered off, and the filtrate was washed with
brine and dried over Na.sub.2SO.sub.4. The solvent was removed to
give the residue (72.45 g) as a white solid. The solid was
dissolved in 80% HOAc aq. solution (400 mL). The mixture was
stirred at R.T. for 15 hours. The mixture was diluted with EtOAc
and washed with NaHCO.sub.3 solution and brine. The organic layer
was dried over Na.sub.2SO.sub.4 and purified by silica gel column
chromatography (1-2% MeOH in DCM) to give P3-4 (37.6 g, 84.2%) as a
white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 7.76 (d,
J=4.0 Hz, 1H), 7.70 (dd, J.sub.1=1.6 Hz, J.sub.2=8.0 Hz, 2H),
7.66.about.7.64 (m, 2H), 7.48.about.7.37 (m, 6H), 6.12 (dd,
J.sub.1=2.8 Hz, J.sub.2=16.8 Hz, 1H), 5.22 (d, J=8.0 Hz, 1H).
5.20.about.5.05 (m, 1H), 4.74 (dd, J.sub.1=5.6 Hz, J.sub.2=17.6 Hz,
1H), 4.16 (d, J=12.0 Hz, 1H), 3.87.about.3.80 (m, 2H), 3.56 (d,
J=12.0 Hz, 1H), 1.16 (s, 9H), 0.92 (s, 9H), 0.14 (s, 6H).
[0253] Preparation of (P3-5): To a solution of P3-4 (11.8 g, 18.8
mmol) in anhydrous DCM (100 mL) was added Dess-Martin periodinane
(16.3 g, 37.6 mmol) at 0.degree. C. under nitrogen. The reaction
was stirred R.T. for 2.5 hours. Water (100 mL) was added, and the
mixture was then filtered. The filtrate was washed with saturated
aq. NaHCO.sub.3 and concentrated. The crude residue was purified by
silica gel column chromatography (20% EtOAc in hexane) to give P3-5
as a white solid (10.1 g, 86.0%).
[0254] Preparation of (P3-6): To a mixture of
methyltriphenylphosphonium bromide (15.7 g, 48.5 mmol) in anhydrous
THF (100 mL) was added n-BuLi (19.4 mL, 48.48 mmol) at -78.degree.
C. under nitrogen. The reaction was stirred at 0.degree. C. for 30
minutes. A solution of P3-5 (10.1 g, 16.2 mmol) in anhydrous THF
(70 mL) was added dropwise at 0.degree. C. under nitrogen. The
reaction was stirred at R.T. for 1.5 hours. The reaction was
quenched by NH.sub.4Cl and extracted with EtOAc. The crude product
was purified by silica gel column chromatography (20% EtOAc in
hexane) to give P3-6 as a white solid (8.3 g, 82.2%). .sup.1H NMR
(CDCl.sub.3, 400 MHz) .delta. 8.16 (s, 1H), 8.81 (d, J=8.0 Hz, 1H),
7.58-7.67 (m, 4H), 7.37-7.46 (m, 6H), 6.17 (d, J=16.0 Hz, 1H), 5.91
(dd, J.sub.1=10.8 Hz, J.sub.2=17.6 Hz, 1H), 5.42 (d, J=17.6 Hz,
1H), 5.22-5.30 (m, 2H), 4.60-4.84 (m, 2H), 3.69 (dd, J.sub.1=11.6
Hz, J.sub.2=21.2 Hz, 2H), 1.10 (s, 9H), 0.91 (s, 1H), 0.12 (d,
J=8.0 Hz, 6H).
[0255] Preparation of (P3-7): To a solution of P3-6 (6.3 g, 10.09
mmol) in anhydrous CH.sub.3CN (50 mL) were added TPSCl (6.1 g, 20.2
mmol), DMAP (2.5 g, 20.2 mmol) and NEt.sub.3 (3 mL) at R.T. The
reaction was stirred at R.T. for 2 hours. NH.sub.4OH (25 mL) was
added, and the reaction was stirred for 1 hour. The mixture was
diluted with DCM (150 mL) and washed with water, 0.1 M HCl and
saturated aq. NaHCO.sub.3. The solvent was removed, and the crude
product was purified by silica gel column chromatography (2% MeOH
in DCM) to give P3-7 as a yellow solid (5.9 g, 93.6%).
[0256] Preparation of (P3-8): To a solution of P3-7 (5.9 g, 9.5
mmol) in MeOH (10 mL) was added Pd/C (1.5 g) at R.T. The reaction
was stirred at R.T. for 2 hours under H.sub.2 (balloon). The
mixture was filtered, and the filtrate was concentrated in vacuo to
give P3-8 as a white solid (5.4 g, 91.3%).
[0257] Preparation of (3a): To a solution of P3-8 (5.4 g, 8.6 mmol)
in MeOH (60 mL) was added NH.sub.4F (10.0 g), and the reaction
mixture was refluxed overnight. After cooling to R.T., the mixture
was filtered, and the filtrate was concentrated. The crude product
was purified by silica gel column chromatography (10% MeOH in DCM)
to give compound 3a as a white solid (1.6 g, 67.8%). .sup.1H NMR
(CD.sub.3OD, 400 M Hz) .delta. 8.08 (d, J=7.6 Hz, 1H), 6.07 (dd,
J.sub.1=3.2 Hz, J.sub.2=15.6 Hz, 1H), 5.88 (d, J=7.2 Hz, 1H), 5.04
(ddd, J=3.2 Hz, J.sub.2=5.2 Hz, J.sub.3=54.0 Hz, 1H), 4.45 (dd,
J=5.2 Hz, J.sub.2=17.2 Hz, 1H), 3.76 (d, J=12.0 Hz, 1H), 3.57 (d,
J=12.0 Hz, 1H), 1.78-1.85 (m, 1H), 1.58-1.67 (m, 1H), 0.95 (t,
J=7.6 Hz, 3H); ESI-MS: m/z 274 [M+H].sup.+, 547 [2M+H].sup.+.
Example 4
Preparation of Compound (4a)
##STR00165##
[0259] To a solution of P3-7 (280 mg, 0.45 mmol) in MeOH (10 mL)
was added NH.sub.4F (1.0 g) at R.T. The reaction mixture was
refluxed for 5 hours. After cooling to R.T., the mixture was
filtered, and the filtrate was concentrated. The crude product was
purified by silica gel column chromatography (10% MeOH in DCM) to
give compound 4a as a white solid (82 mg, 67.2%1.6 g, 67.8%).
.sup.1H NMR (CD.sub.3OD, 400 M Hz) .delta. 8.11 (d, J=7.6 Hz, 1H),
5.99-6.08 (m, 2H), 5.88 (d, J=7.6 Hz, 1H), 5.47 (dd, J.sub.1=1.2
Hz, J.sub.2=17.2 Hz, 1H), 5.26 (dd, J.sub.1=1.6 Hz, J.sub.2=11.2
Hz, 1H), 4.97 (d, J=5.2 Hz, 0.5H), 4.82 (d, J=7.6 Hz, 0.5H), 4.52
(dd, J.sub.1=5.2 Hz, J.sub.2=23.2 Hz, 1H), 3.65 (d, J=12.4 Hz, 1H),
3.54 (d, J=12.4 Hz, 1H); ESI-MS: m/z 272 [M+H].sup.+, 543
[2M+H].sup.+.
Example 5
Preparation of Compound (5a)
##STR00166##
[0261] Preparation of (P5-1): To a solution of P3-6 (600 mg, 0.96
mmol) in MeOH (30 mL) was added 10% Pd/C (320 mg) at R.T. The
mixture was stirred under H.sub.2 balloon at R.T. for 3 hours. The
reaction mixture was filtered, and the filtrate was concentrated to
give P5-1 (540 mg, 89.8%) as a colorless solid. The crude product
was used directly for the next step without purification.
[0262] Preparation of (5a): To a solution of P5-1 (540 mg, 0.86
mmol) in MeOH (8 mL) was added NH.sub.4F (1.2 g, 32.4 mmol) R.T.
The mixture was refluxed for 30 hours. The solid was removed by
filtration, and the filtrate was concentrated. The residue was
purification by silica gel column chromatography (2.5%-9% MeOH in
DCM) to give compound 5a (190 mg, 80.6%) as a colorless solid.
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.05 (d, J=8.0 Hz, 1H),
6.09 (dd, J.sub.1=4.0 Hz, J.sub.2=14.8 Hz, 1H), 5.04-5.20 (m, 1H),
4.42 (dd, J.sub.1=5.2 Hz, J.sub.2=13.6 Hz, 1H), 3.71 (d, J=11.6 Hz,
1H), 3.57 (d, J=12.0 Hz, 1H), 1.61-1.82 (m, 2H), 0.94 (t, J=7.2 Hz,
3H).
Example 6
Preparation of Compound (6a)
##STR00167##
[0264] Preparation of (P6-1): To a solution of P3-3 (800 mg, 2.05
mmol) in anhydrous DCM (15 mL) were added imidazole (558 mg, 8.2
mmol), TBSCl (1.2 g, 8.2 mmol) and AgNO.sub.3 (700 mg, 4.1 mmol) at
R.T. The reaction mixture was stirred at R.T. overnight. The
mixture was filtered, and the filtrate was washed with brine and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel to give P6-1 as a white solid (950 mg,
79.2%).
[0265] Preparation of (6a): To a solution of P6-1 (600 mg, 0.97
mmol) in anhydrous CH.sub.3CN (18 mL) was added DMAP (239 mg, 2.91
mmol), NEt.sub.3 (294 mg, 2.91 mmol) and TPSCl (879 mg, 2.91 mmol)
at R.T. The reaction was stirred at R.T. for 1 hour. NH.sub.4OH (9
mL) was added, and the reaction was stirred for 3 hours. The
mixture was diluted with EtOAc (200 mL) and washed with water, 0.1
M HCl and saturated aq. NaHCO.sub.3. The organic layer was
separated, dried and concentrated to give a crude residue. The
crude residue was purified by column chromatography on silica gel
to give the product as a white solid (500 mg, 83.3%). The solid was
treated with NH4F (1.0 g) in MeOH (20 mL) at refluxed temperature
for 5 hours. The mixture was filtered, and the filtrate was
concentrated in vacuo. The residue was purified by column
chromatography on silica gel (15% MeOH in DCM) to give compound 6a
as a white solid (132 mg, 59.3%). .sup.1H NMR (DMSO-d6, 400 MHz)
.delta. 7.89 (d, J=7.6 Hz, 1H), 7.22 (d, J=18.8 Hz, 2H), 6.09 (dd,
J.sub.1=4.4 Hz, J.sub.2=14.8 Hz, 1H), 5.73 (d, J=5.2 Hz, 1H), 5.52
(d, J=5.6 Hz, 1H), 5.12 (t, J=4.8 Hz, 1H), 4.90-5.06 (m, 1H), 4.50
(t, J=6.0 Hz, 1H), 4.27-4.33 (m, 1H), 3.66 (dd, J.sub.1=5.2 Hz,
J.sub.2=12.0 Hz, 1H), 3.47-3.58 (m, 3H); ESI-MS: m/z 276
[M+H].sup.+, 551 [2M+H].sup.+.
Example 7
Preparation of Compound (7a)
##STR00168##
[0267] Preparation of (P7-1): A mixture of P3-4 (1.60 g, 2.5 mmol),
PPh.sub.3 (1.3 g, 5.0 mmol) and CCl.sub.4 (0.76 g, 5.0 mmol) in DCE
(20 mL) was heated to 130.degree. C. under microwave irradiation
under N.sub.2 for 40 mins. After cooled to R.T., the solvent was
removed, and the residue was purified on a silica gel column
(PE/EA=50/1 to 10/1) to give P7-1 (1.1 g, 68.8%) as a white
solid.
[0268] Preparation of (P7-2): P7-1 (0.80 g, 1.3 mmol), DMAP (0.3 g,
2.6 mmol), TPSCl (0.8 g, 2.6 mmol) and Et.sub.3N (0.3 g, 2.6 mmol)
were dissolved in MeCN (30 mL). The mixture was stirred at R.T. for
14 hours. NH.sub.3 in THF (saturated at 0.degree. C., 100 mL) was
added to the mixture, and the mixture was stirred at R.T. for 2
hours. The solvent was removed, and the residue was purified by
column (DCM/MeOH=100:1 to 50:1) to give P7-2 (0.63 g, 78.8%) as a
white solid.
[0269] Preparation of (7a): To a solution of P7-2 (0.63 g, 0.98
mmol) in MeOH (10 mL) was added NH.sub.4F (0.3 g), and the reaction
was refluxed for 12 hours. The reaction was cooled to R.T., and the
precipitate was filtered off. The filtrate was concentrated in
vacuo. The residue was purified by silica gel column chromatography
(10% MeOH in DCM) to give compound 7a as a white solid (153 mg,
53.5%). .sup.1H NMR (CD.sub.3OD, 400 M Hz) .delta. 8.05 (d, J=7.2
Hz, 1H), 6.14 (dd, J.sub.1=3.6 Hz, J.sub.2=15.2 Hz, 1H), 5.92 (d,
J=7.2 Hz, 1H), 5.15 (ddd, J.sub.1=4.0 Hz, J.sub.2=5.2 Hz,
J.sub.3=53.6 Hz, 1H), 4.57 (dd, J.sub.1=4.8 Hz, J.sub.2=15.2 Hz,
1H), 3.93 (d, J=11.6 Hz, 1H), 3.75-3.84 (m, 3H); ESI-MS: m/z 294
[M+H].sup.+, 587 [2M+H].sup.+.
Example 8
Preparation of Compound (8a)
##STR00169##
[0271] To a solution of P7-1 (630 mg, 0.5 mmol) in MeOH (10 mL) was
added NH.sub.4F (0.1 g), and the reaction was refluxed for 12
hours. The mixture was filtered, and the filtrate was concentrated
in vacuo. The crude product was purified by silica gel column
chromatography (10% MeOH in DCM) to give compound 8a as a white
solid (153 mg, 53.5%). .sup.1H NMR (CD.sub.3OD, 400 M Hz) .delta.
7.99 (d, J=8.0 Hz, 1H), 6.17 (dd, J.sub.1=4.4 Hz, J.sub.2=14.4 Hz,
1H), 5.70 (d, J=8.0 Hz, 1H), 5.22 (ddd, J.sub.1=J.sub.2=4.8 Hz,
J.sub.3=53.2 Hz, 1H), 4.55 (dd, J.sub.1=5.2 Hz, J.sub.2=12.4 Hz,
1H), 3.88 (d, J=12.0 Hz, 1H), 3.76-3.79 (m, 3H); Negative-ESI-MS:
m/z 293 [M-H].sup.-.
Example 9
Preparation of Compound (9a)
##STR00170##
[0273] Preparation of (P9-1): A mixture of P3-4 (3.2 g, 5.0 mmol),
Ph.sub.3P (5.2 g, 20 mmol), iodine (2.60 g, 10.2 mmol) and
imidazole (1.4 g, 20 mmol) in anhydrous THF (40 mL) was stirred at
80.degree. C. for 14 hours. The reaction was cooled to R.T. and
quenched with saturated aq. Na.sub.2S.sub.2O.sub.3. The solution
was extracted with EA. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by
silica gel column chromatography (20-50% EA in PE) to give P9-1
(1.6 g, 68.2%) as a white solid.
[0274] Preparation of (P9-2): A mixture of P9-1 (1.4 g, 0.2 mmol),
Et.sub.3N (40 mg, 0.4 mmol) and Pd/C in EtOH (20 mL) was stirred at
R.T. under H.sub.2 (balloon) overnight. The precipitate was
filtered off, and the filtrate was concentrated. The residue was
purified on a silica gel column (20%-50% EtOAc in PE) to give P9-2
as a white solid (1.1 g, 78%). H NMR (CDCl.sub.3, 400 MHz) .delta.
8.11 (br s, 1H), 7.76 (d, J=8.0 Hz, 1H), 7.39-7.67 (m, 10H), 6.18
(dd, J.sub.1=3.2 Hz, J.sub.2=14.4 Hz, 1H), 5.26-5.30 (m, 1H), 4.86
(m, 1H), 4.42 (dd, J.sub.1=5.2 Hz, J.sub.2=15.2 Hz, 1H), 3.81 (d,
J=11.2 Hz, 1H), 3.58 (d, J=11.2 Hz, 1H), 1.16 (s, 3H), 1.11 (s,
9H), 0.91 (s, 9H), 0.13 (s, 3H), 0.08 (s, 3H).
[0275] Preparation of (P9-3): P9-2 (650 mg, 1.1 mmol), DMAP (270
mg, 2.2 mmol), TPSCl (664 mg, 2.2 mol) and Et.sub.3N (222 mg, 2.2
mmol) were dissolved in MeCN (20 mL). The mixture was stirred at
R.T. for 14 hours. The reaction was added NH.sub.3 in THF
(saturated at 0.degree. C.), and the mixture was stirred at R.T.
for 2 hours. The solvent was removed, and the residue was purified
on a silica gel column (1-10% MeOH in DCM) to give P9-3 (430 mg,
crude) as a light yellow syrup.
[0276] Preparation of (9a): A mixture of P9-3 (430 mg, 0.7 mmol)
and NH.sub.4F (97 mg, 2.1 mmol) in MeOH (10 mL) was refluxed for 14
hours. The solvent was removed, and the residue was purified on a
silica gel column (5%-10% MeOH in DCM) to give compound 9a as a
white solid (64.8 mg, 35.4%). .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 8.10 (d, J=7.6 Hz, 1H), 6.03 (dd, J.sub.1=2.0 Hz,
J.sub.2=16.8 Hz, 1H), 5.87 (d, J=7.6 Hz, 1H), 4.98 (m, 1H), 4.37
(dd, J.sub.1=5.2 Hz, J.sub.2=21.6 Hz, 1H), 3.59 (dd, J.sub.1=12.0
Hz, J.sub.2=28.4 Hz, 2H), 1.23 (d, J=0.8 Hz, 3H).
Example 10
Preparation of Compound (10a)
##STR00171##
[0278] To a stirred solution of P9-2 (400 mg, 0.65 mmol) in MeOH
(20 mL) was added NH.sub.4F (52 mg, 1.5 mmol). The mixture was
refluxed overnight. The solvent was removed, and the residue was
purified on a silica gel column (5-10% MeOH in DCM) to give
compound 10a (140 mg, 82.4%) as a white solid. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.05 (d, J=8.4 Hz, 1H), 6.06 (dd,
J.sub.1=2.8 Hz, J.sub.2=16.4 Hz, 1H), 5.67 (d, J=8.0 Hz, 1H), 5.08
(m, 1H), 4.37 (d, J.sub.1=5.2 Hz, J.sub.2=18.8 Hz, 1H), 3.59 (dd,
J.sub.1=12.0 Hz, J.sub.2=26.4 Hz, 2H), 1.23 (s, 3H). ESI-TOF-MS:
m/z 283 [M+Na].sup.+.
Example 11
Preparation of Compound (11a)
##STR00172##
[0280] Preparation of (P11-1): To a solution of P3-5 (2.1 g, 3.5
mmol) in anhydrous THF (25 mL) was added ethynylmagnesium bromide
(5.1 mmol) at -78.degree. C. The reaction was stirred at 0.degree.
C. for 3 hours. The reaction was quenched with saturated aq.
NH.sub.4Cl (10 mL). The mixture was diluted with EtOAc (200 mL) and
washed with water and brine. The organic layer was dried and
concentrated to give a residue. The residue was purified by column
chromatography on silica gel (eluting with DCM:MeOH=60:1) to give
P11-1 as a white solid (870 mg, 83.3%).
[0281] Preparation of (P11-2): P11-1 (870 mg, 1.34 mmol) was
dissolved in anhydrous DCM (12 mL), and methyl chloroformate (2.3
mL) and pyridine (2.5 mL) were added at R.T. The reaction mixture
was stirred at R.T. for 1 hour. The mixture was diluted with DCM
and washed with saturated aq. NaHCO.sub.3. The organic layer was
separated, dried and concentrated to give a residue. The residue
was purified by column chromatography on silica gel (eluting with
PE:EtOAc=8:1) to give a crude product as a white solid (830 mg,
88.4%). To a mixture of Pd.sub.2(dba).sub.3 (55 mg, 0.06 mmol) in
anhydrous DMF (12 mL) was added P(nBu).sub.3 (35 mg, 0.17 mmol) and
HCOONH.sub.4 (108 mg, 1.7 mmol) at R.T. under nitrogen. The
reaction mixture was stirred at R.T. for 30 min. A solution of the
crude product (830 mg, 1.16 mmol) in anhydrous DMF (16 mL) was
added, and the reaction mixture was stirred at 70.degree. C. for 3
hours. The reaction was diluted with EtOAc and washed with brine.
The organic layer was separated, dried and concentrated to give a
residue. The residue was purified by column chromatography on
silica gel (eluting with PE:EtOAc=9:1) to give P11-2 as a white
solid (510 mg, 67.6%). .sup.1H NMR (CD.sub.3OD, 400 M Hz) .delta.
7.61-7.75 (m, 5H), 7.36-7.47 (m, 6H), 6.04 (d, J=18.8 Hz, 1H), 5.34
(t, J=6.8 Hz, 1H), 5.21 (dd, J.sub.1=1.2 Hz, J.sub.2=7.2 Hz, 1H),
5.10 (q, J.sub.1=5.2 Hz, J.sub.2=53.6 Hz, 1H), 4.80-4.92 (m, 1H),
4.59-4.79 (m, 2H), 3.86 (d, J=12.0 Hz, 1H), 3.75 (d, J=12.0 Hz,
1H), 1.09 (s, 9H), 0.92 (d, J=4.4 Hz, 9H), 0.15 (t, J=4.0 Hz,
6H).
[0282] Preparation of (P11-3): To a solution of P11-2 (490 mg, 0.77
mmol) in anhydrous MeCN (15 mL) was added TPSCl (700 mg, 2.31
mmol), DMAP (282 mg, 2.31 mmol) and TEA (234 mg, 2.31 mmol) at R.T.
The reaction mixture was stirred at room temperature for 1 hour.
Then NH.sub.4OH (8 mL) was added and the reaction mixture was
stirred for another 4 hours. The mixture was diluted with EtOAc and
washed with water, 1.0 M aq. HCl and saturated aq. NaHCO.sub.3. The
organic layer was separated and dried, concentrated to give the
residue which was purified by HPLC separation (MeCN and 0.1% HCOOH
in water) to give P11-3 as a white solid (190 mg, 38.8%). .sup.1H
NMR (CD.sub.3OD, 400 MHz) .delta. 7.88 (d, J=7.2 Hz, 1H), 7.63-7.70
(m, 4H), 7.37-7.48 (m, 6H), 6.12 (d, J=18.4 Hz, 1H), 5.49 (d, J=7.6
Hz, 1H), 5.34 (t, J=6.8 Hz, 1H), 4.84-5.01 (m, 2H), 4.66-4.78 (m,
2H), 3.89 (d, J=11.6 Hz, 1H), 3.75 (d, J=11.6 Hz, 1H), 1.10 (s,
9H), 0.91 (d, J=3.2 Hz, 9H), 0.13 (t, J=5.2 Hz, 6H).
[0283] Preparation of (11a): To a solution of P11-3 (130 mg, 0.21
mmol) in MeOH (8 mL) was added NH.sub.4F (1 g), and the reaction
mixture was refluxed for 6 hours. The mixture was filtered, and the
filtrate was concentrated in vacuo. The residue was purified by
column chromatography on silica gel (eluting with DCM:MeOH=13:1) to
give compound 11a as a white solid (47 mg, 79.1%). .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.07 (d, J=7.6 Hz, 1H), 6.05 (dd,
J.sub.1=1.2 Hz, J.sub.2=16.8 Hz, 1H), 5.86 (d, J=7.6 Hz, 1H), 5.40
(dd, J.sub.1=J.sub.2=6.8 Hz, 1H), 4.87-4.99 (m, 3H), 4.46-4.80 (m,
1H), 3.75 (d, J=12.4 Hz, 1H), 3.68 (d, J=12.4 Hz, 1H); ESI-MS: m/z
284.02 [M+H].sup.+, 567.08 [2M+H].sup.+.
Example 12
Preparation of Compound (12a)
##STR00173##
[0285] Preparation of (P12-1): To a solution of P3-4 (500 mg, 0.8
mmol) in anhydrous toluene (12 mL) was added DAST (0.3 mL, 2 mmol)
at -65.degree. C. under nitrogen. The reaction mixture was stirred
at R.T. for 2 hours. The reaction was quenched with saturated aq.
NaHCO.sub.3 and extracted with EtOAc. The organic layer was
separated, dried and concentrated to give the residue. The residue
was purified by column chromatography on silica gel (eluting with
PE:EtOAc=9:1) to give P12-1 as a yellow solid (170 mg, 42.5%).
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 7.66 (dd, J.sub.1=1.6 Hz,
J.sub.2=18.0 Hz, 4H), 7.54 (d, J=7.6 Hz, 1H), 7.35-7.47 (m, 6H),
6.59 (dd, J.sub.1=5.6 Hz, J.sub.2=14.0 Hz, 1H), 5.78 (d, J=7.6 Hz,
1H), 5.05-5.24 (m, 2H), 4.93 (d, J=7.6 Hz, 1H), 4.57 (d, J=7.6 Hz,
1H), 3.93-4.00 (m, 2H), 1.07 (d, J=2.4 Hz, 9H).
[0286] Preparation of (P12-2): To a solution of P12-1 (100 mg, 0.2
mmol) in anhydrous MeCN (5 mL) was added TPSCl (182 mg, 0.6 mmol),
DMAP (68 mg, 0.6 mmol) and TEA (61 mg, 0.6 mmol) at R.T. under
nitrogen. The reaction mixture was stirred at R.T. for 1 hour.
NH.sub.4OH (3 mL) was added, and the reaction was stirred for 2
hours. The mixture was diluted with EtOAc and washed with water,
1.0 M HCl and saturated aq. NaHCO.sub.3. The organic layer was
separated, dried and concentrated to give a residue. The residue
was purified by column chromatography on silica gel (DCM:MeOH=50:1)
to give P12-2 as a yellow solid (96 mg, 96%).
[0287] Preparation of (12a): To a solution of P12-2 (96 mg, 0.2
mmol) in MeOH (5 mL) was added NH.sub.4F (500 mg) at R.T. The
reaction was refluxed for 3 hours. The mixture was filtered, and
the residue was purified by RP HPLC (MeCN and 0.1% HCOOH in water)
to give compound 12a as a white solid (25 mg, 48.7%). .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 7.85 (d, J=7.6 Hz, 1H), 6.59 (dd,
J.sub.1=5.2 Hz, J.sub.2=12.8 Hz, 1H), 6.04 (d, J=7.6 Hz, 1H),
5.10-5.26 (m, 2H), 4.79-4.90 (m, 1H), 4.57 (d, J=7.6 Hz, 1H), 3.82
(d, J=12.4 Hz, 1H), 3.76 (dd, J.sub.1=1.6 Hz, J.sub.2=12.4 Hz, 1H);
ESI-MS: m/z 257.9 [M+H].sup.+, 514.8 [2M+H].sup.+.
Example 13
Preparation of Compound (13a)
##STR00174##
[0289] Preparation of (P13-1): To a solution of compound 3a (700
mg, 2.56 mmol) in anhydrous pyridine (5 mL) were added TBDPSCl (2.8
g, 10.24 mmol), imidazole (522 mg, 7.68 mmol) and AgNO.sub.3 (870
mg, 5.12 mmol) at R.T. under N.sub.2. The reaction mixture was
stirred at R.T. for 3 hours. The mixture was diluted with MeOH and
filtered. The mixture was concentrated, and the residue was
purified by column chromatography on silica gel (eluting with
DCM:MeOH=80:1.about.40:1) to give the crude intermediate as a
yellow solid (1.05 g, 80.8%). .sup.1H NMR (DMSO-d6, 400 MHz)
.delta. 7.75 (d, J=7.6 Hz, 1H), 7.61-7.65 (m, 4H), 7.41-7.50 (m,
7H), 6.02 (dd, J.sub.1=2.8 Hz, J.sub.2=17.2 Hz, 1H), 5.69 (d, J=6.0
Hz, 1H), 5.56 (d, J=7.6 Hz, 1H), 4.96-5.11 (m, 1H), 4.37-4.46 (m,
1H), 3.82 (d, J=10.8 Hz, 1H), 3.62 (d, J=10.8 Hz, 1H), 1.70-1.78
(m, 1H), 1.53-1.59 (m, 1H), 1.02 (s, 9H), 0.79 (t, J=7.6 Hz, 3H).
To a solution of the crude intermediate (1.0 g, 1.96 mmol) in
anhydrous DCM (15 mL) were added sym-collidine (1.4 g, 11.76 mmol),
AgNO.sub.3 (1.0 g, 5.88 mmol) and MMTrC (4.8 g, 15.6 mmol) at R.T.
under N.sub.2. The reaction mixture was stirred at R.T. overnight.
The mixture was filtered and concentrated. The residue was purified
by column chromatography on silica gel (eluting with PE:EtOAc=2:1)
to give crude full protected intermediates as a white solid (1.1 g,
53.1%). To a solution of the crude intermediate (600 mg, 0.57 mmol)
in THF (5 mL) was added TBAF (446 mg, 1.71 mmol)) at R.T. The
reaction was stirred at 40-50.degree. C. overnight. The crude
product was purified by column chromatography on silica gel eluted
with PE:EtOAc=3:2 to give crude P13-1 (350 mg, 75.1%) as a yellow
solid.
[0290] Preparation of (13a): To a solution of P13-1 (300 mg, 0.37
mmol) in CH.sub.3CN (2.5 mL) were added NMI (2.5 mL) and a solution
of phenyl(isopropoxy-L-alaninyl) phosphorochloridate (2.55 g, 7.4
mmol) in CH.sub.3CN (2.5 mL) at R.T. under N.sub.2. The reaction
mixture was stirred at R.T. for 3 hours. The mixture was
concentrated in vacuo. The residue was purified by column
chromatography on silica gel (PE:EtOAc=1:1) to give crude product
as a yellow oil (500 mg, 81%). The crude product was further
treated with 80% HCOOH (70 mL) at R.T. overnight. The mixture was
concentrated in vacuo, and the crude product was purified by RP
HPLC (MeCN and 0.1% HCOOH in water) to give compound 13a as a white
solid (a mixture of two P isomers, 86 mg, 40.3% two steps). .sup.1H
NMR (CD.sub.3OD, 400 MHz) .delta. 7.75, 7.71 (2d, J=7.6 Hz, 1H),
7.33-7.38 (m, 2H), 7.19-7.26 (m, 3H), 6.02-6.10 (m, 1H), 5.87, 5.82
(2d, J=7.6 Hz, 1H), 4.99-5.02 (m, 0.5H), 4.72-4.82 (m, 1.5H),
4.14-4.43 (m, 3H), 3.89-3.94 (m, 1H), 1.68-1.81 (m, 6H), 1.51-1.56
(m, 1H), 1.30-1.43 (m, 8H), 0.96-1.01 (m, 3H); ESI-MS: m/z 582.93
[M+H].sup.+.
Example 14
Preparation of Compound (14a)
##STR00175##
[0292] Preparation of (P14-1): To a stirred solution of P13-1 (451
mg, 0.55 mmol) and NMI (1 mL) in anhydrous acetonitrile (2 mL) was
added dropwise a solution of
2-chloro-8-methyl-4H-benzo[d][1,3,2]dioxaphosphinine (855 mg, 4.2
mmol) in acetonitrile (0.2 mL) at 0.degree. C. under N.sub.2. The
mixture was stirred at R.T. for 2 hours. Solution of I.sub.2 (3.2
g, 12.6 mmol), pyridine (9 mL), H.sub.2O (3 mL) and DCM (3 mL) was
added. The reaction mixture was stirred for 30 mins. The reaction
was quenched with NaS.sub.2O.sub.3 solution and extracted with EA.
The organic layer was dried over Na.sub.2SO.sub.4 and concentrated.
The residue was purified by column on silica gel (PE:EA=1:1 to 1:2)
to give P14-1 (205 mg, 37%) as a white solid.
[0293] Preparation of (14a): P14-1 (205 mg, 0.21 mmol) was
dissolved in 80% HCOOH aq. solution, and the mixture was stirred at
R.T. for 16 hours. The solvent was removed, and the residue was
purified by RP HPLC (HCOOH system) to give compound 14a as a
mixture of 2 P-isomers (24 mg, 18%). .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 7.60, 7.53 (2d, J=8.0 Hz, 1H), 7.21-7.25 (m, 1H),
7.02-7.12 (m, 2H), 5.95.5.87 (2dd, J.sub.1=2.4 Hz, J.sub.2=18.0 Hz,
1H), 5.71, 5.69 (2d, J=8.0 Hz, 1H), 5.38-5.53 (m, 2H), 5.06, 5.04
(2ddd, J.sub.1=2.4 Hz, J.sub.2=5.6 Hz, J.sub.3=54.0 Hz, 1H),
4.32-4.49 (m, 2H), 2.26 (d, J=3.6 Hz, 3H), 1.83-1.92 (m, 1H),
1.64-1.72 (m, 1H), 0.96, 0.93 (2t, J=7.6 Hz, 3H). .sup.31P NMR
(CD.sub.3OD, 162 MHz) .delta. -8.22, -8.50; ESI-LCMS: m/z 456
[M+H].sup.+.
Example 15
Preparation of Compound (15a)
##STR00176##
[0295] Step 1. Preparation of (P15-1): To a mixture of P3-8 (2.2 g,
2.5 mmol), AgNO.sub.3 (844 mg, 5.0 mmol) and collidine (907 mg, 7.5
mmol) in anhydrous DCM (10 mL) was added MMTrCl (1.54 g, 5.0 mmol)
under N.sub.2. The reaction mixture was stirred at R.T. overnight.
The reaction mixture was filtered through a Buchner Funnel. The
filtrate was washed with saturated NaHCO.sub.3 solution and brine.
The organic layer was separated, dried over anhydrous
Na.sub.2SO.sub.4 and filtered. The filtrate was concentrated to
dryness. The residue was purified by column on silica gel
(PE:EA=10:1 to 1:2) to give the intermediate (2.3 g, 84%), which
was dissolved in a solution of TBAF in THF (1M, 2.6 mL) under
N.sub.2. The reaction mixture was stirred at R.T. overnight. The
residue was dissolved in EA (200 mL) and washed with water and
brine. The organic layer was separated, dried over anhydrous
Na.sub.2SO.sub.4 and filtered. The filtrate was concentrated to
dryness, and the residue was purified by column on silica gel
(DCM/MeOH=100:1 to 30:1) to give P15-1 as a white foam (1.3 g,
94%).
[0296] Preparation of (15a): To a stirred solution of P15-1 (300
mg, 0.55 mmol) and proton sponge (235 mg, 1.1 mmol) in anhydrous
MeCN (9 mL) was added with a solution of POCl.sub.3 (169 mg, 1.1
mmol) in MeCN (1 mL) via syringe at 0.degree. C. The mixture was
stirred at R.T. for 40 mins. A mixture of (S)-cyclohexyl
2-aminopropanoate hydrochloride (525 mg, 2.55 mmol) and TEA (0.1
mL) was added at 0.degree. C. The mixture was warmed to R.T. and
stirred for 3 hours. The reaction mixture was quenched with
saturated NaHCO.sub.3, and extracted with EA (100 mL.times.2). The
combined organic layers was dried over Na.sub.2SO.sub.4,
concentrated and purified by silica gel column (1-4% MeOH in DCM)
to give the crude product (400 mg, 78.15%) as a yellow solid. The
crude product was treated with 80% HCOOH (50 mL) at R.T. for 16
hours. The solvent was removed, and the residue was purified by RP
HPLC to give compound 15a as a white solid (40 mg, 14%). .sup.1H
NMR (MeOD, 400 MHz) .delta. 7.82 (d, J=7.6 Hz, 1H), 6.09 (dd,
J.sub.1=2.8 Hz, J.sub.2=14.0 Hz, 1H), 5.98 (d, J=7.6 Hz, 1H), 5.04
(ddd, J.sub.1=3.2 Hz, J.sub.2=5.6 Hz, J.sub.3=53.6 Hz, 1H),
4.71-4.77 (m, 2H), 4.45 (dd, J.sub.1=5.6 Hz, J.sub.2=12.4 Hz, 1H),
4.14-4.18 (m, 1H), 3.97-4.01 (m, 1H), 3.84-3.92 (m, 2H), 1.31-1.87
(m, 28H), 0.99 (t, J=7.2 Hz, 3H). .sup.31P NMR (CD.sub.3OD, 162
MHz) .delta. 13.94; ESI-LCMS: m/z 660 [M+H].sup.+.
Example 16
Preparation of Compound (16a)
##STR00177##
[0298] To a stirred solution of compound 4a (150 mg, 0.56 mmol) in
anhydrous THF (3 mL) was added dropwise a solution of t-BuMgCl (1.2
mL, 1M in THF) at -78.degree. C. The mixture was stirred at
0.degree. C. for 30 min and re-cooled to -78.degree. C. A solution
of phenyl(isopropoxy-L-alaninyl) phosphorochloridate (312 mg, 1.2
mmol) in THF (1.0 mL) was added dropwise. After addition, the
mixture was stirred at 25.degree. C. for 16 hours. The reaction was
quenched with HCOOH (80% aq.) at 0.degree. C. The solvent was
removed, and the residue was purified on silica gel (DCM:MeOH=50:1
to 10:1) to give compound 16a as a white solid (24.0 mg, 15%).
.sup.1H NMR (MeOD, 400 MHz) .delta. 7.76 (d, J=7.2 Hz, 1H),
7.17-7.38 (m, 5H), 6.01-6.08 (m, 2H), 5.81 (d, J=7.6 Hz, 1H),
5.54-5.58 (m, 1H), 5.35-5.38 (m, 1H), 4.92-4.97 (m, 2H), 4.45-4.52
(m, 1H), 4.08-4.19 (m, 2H), 3.88-3.92 (m, 1H), 1.28-1.33 (m, 3H),
1.20-1.22 (m, 6H); .sup.31P NMR (CD.sub.3OD, 162 MHz) .delta. 7.36;
ESI-LCMS: m/z 541.0[M+H].sup.+.
Example 17
Preparation of Compound (17a)
##STR00178##
[0300] Preparation of (P17-1): To a solution of P3-7 (1.4 g, 2.3
mmol) in MeOH (50 mL) was added NH.sub.4F (8.0 g) at R.T. The
reaction mixture was refluxed overnight. After cooling to R.T., the
mixture was filtered, and the filtrate was concentrated. The crude
product was purified by silica gel column chromatography (10% MeOH
in DCM) to give P17-1 as a white solid (410 mg, 77.8%).
[0301] Preparation of (P17): To a stirred solution of P17-1 (60 mg,
0.19 mmol) in anhydrous THF (3 mL) was added dropwise a solution of
t-BuMgCl (0.38 mL, 1M in THF) at -78.degree. C. The mixture was
stirred at 0.degree. C. for 30 min and re-cooled to -78.degree. C.
A solution of phenyl(isopropoxy-L-alaninyl) phosphorochloridate
(104 mg, 0.4 mmol) in THF (0.5 mL) was added dropwise. After
addition, the mixture was stirred at 25.degree. C. for 16 hours.
The reaction was quenched with HCOOH (80% aq.) at 0.degree. C. The
solvent was removed, and the residue was purified on silica gel
(DCM:MeOH=50:1 to 10:1) to give compound 17a as a white solid (a
mixture of two P isomers, 11.0 mg, 11%). .sup.1H NMR (MeOD, 400
MHz) .delta. 7.71 (2d, J=8.0 Hz, 1H), 7.17-7.37 (m, 5H), 5.98-6.07
(m, 2H), 5.61, 5.68 (2d, J=8.0 Hz, 1H), 5.53-5.58 (m, 1H),
5.35-5.40 (m, 1H), 5.08-5.10 (m, 1H), 4.93-4.99 (m, 1H), 4.52-4.53
(m, 1H), 4.16-4.21 (m, 1H), 4.06-4.11 (m, 1H), 3.86-3.94 (m, 1H),
1.28-1.34 (m, 3H), 1.20-1.22 (m, 6H). .sup.31P NMR (MeOD, 162 MHz)
.delta. 3.72, 3.45. ESI-LCMS: m/z 542.0 [M+H].sup.+.
Example 18
Preparation of Compound (18a)
##STR00179##
[0303] Preparation of (P18-1): To a solution of
(chloromethyl)triphenylphosphonium chloride (2.1 g, 6.0 mmol) in
anhydrous THF (10 mL) was added dropwise n-BuLi (4.6 mL, 6.0 mmol)
at -70.degree. C. under nitrogen. The reaction was stirred at
-70.degree. C. for 50 mins. A solution of compound P3-9 (950 mg,
1.5 mmol) in anhydrous THF (5 mL) was added at -70.degree. C., and
the reaction was stirred at 0.degree. C. for 3 hours. The reaction
was quenched by saturated aq. NH.sub.4Cl and extracted with EtOAc.
The organic layer was separated, dried and concentrated to give a
residue. The residue was purified by column chromatography on
silica gel (eluting with PE:EtOAc=6:1) to give P18-1 as a yellow
gum (900 mg, 91.2%).
[0304] Preparation of (P18-2): To a solution of compound P18-1 (600
mg, 0.91 mmol) in anhydrous THF (18 mL) was added dropwise n-BuLi
(4.7 mL, 10.9 mmol) at -70.degree. C. under nitrogen. The reaction
was stirred at -70.degree. C. for 3 hours. The reaction was
quenched by saturated aq. NH.sub.4Cl and extracted with EtOAc. The
organic layer was separated, dried and concentrated to give a
residue. The residue was purified by column chromatography on
silica gel (eluting with PE:EtOAc=8:1-5:1) to give P18-2 as a white
solid (300 mg, 53.0%).
[0305] Preparation of (P18-3): To a solution of P18-2 (300 mg, 0.44
mmol) in MeOH (10 mL) was added NH.sub.4F (1.0 g) at R.T. The
reaction was refluxed for 3 hours. After cooling R.T., the mixture
was filtered, and the filtrate was concentrated in vacuo. The
residue was purified by column chromatography on silica gel
(eluting with DCM:MeOH=50:1-30:1) to give P18-3 as a white solid
(135 mg, 78.1%). .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 7.84 (d,
J=8.0 Hz, 1H), 6.06 (dd, J.sub.1=1.6 Hz, J.sub.2=19.6 Hz, 1H), 5.67
(d, J=8.4 Hz, 1H), 5.18-5.03 (m, 1H), 4.50 (dd, J.sub.1=5.2 Hz,
J.sub.2=21.6 Hz, 1H), 3.85 (d, J=12.4 Hz, 1H), 3.72 (d, J=12.4 Hz,
1H), 3.09 (s, 1H).
[0306] Preparation of (18a): To a solution of P18-3 (130 mg, 0.5
mmol) in anhydrous THF (4 mL) was added dropwise t-BuMgCl (1.0 mL,
1.0 mmol) at -70.degree. C. under nitrogen. The reaction was
stirred at R.T. for 30 mins. A solution of
phenyl(isopropoxy-L-alaninyl) phosphorochloridate in anhydrous THF
(1M, 0.8 mL, 0.78 mmol) was added at -70.degree. C., and the
reaction mixture was stirred at R.T. for 5 hours. The reaction was
quenched by HCOOH, and the mixture was concentrated in vacuo. The
residue was purified by column chromatography on silica gel
(DCM:MeOH=60:1) to give compound 18a as a white solid (a mixture of
two P isomers, 25 mg, 7.7%). .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 7.64, 7.60 (2d, J=7.6 Hz, 1H), 7.32-7.36 (m, 2H), 7.16-7.25
(m, 3H), 5.95-6.01 (m, 1H), 5.67, 5.62 (2d, J=8.0 Hz, 1H),
5.10-5.25 (m, 1H), 4.93-4.97 (m, 1H), 4.49-4.59 (m, 1H), 4.33-4.42
(m, 1H), 4.24-4.29 (m, 1H), 3.86-3.94 (m, 1H), 3.25, 3.22 (2s, 1H),
1.28-1.34 (m, 3H), 1.20-1.23 (m, 6H); ESI-MS: m/z 540.2
[M+H].sup.+.
Example 19
Preparation of Compound (19a)
##STR00180##
[0308] Preparation of (P19-1): P15-2 (1.2 g, 2.2 mmol) was
dissolved in dry acetonitrile (20 mL), and 0.45 M tetrazole (24.0
mL, 11.0 mmol) and
3-(bis(diisopropylamino)phosphinooxy)propanenitrile (1.13 g, 3.74
mmol) was added. The reaction mixture was stirred for 1 hour under
N.sub.2 at R.T. TBDPH (2.7 mL, 15 mmol) was added, and the mixture
was stirred for 1 hour. The reaction was quenched by
Na.sub.2S.sub.2O.sub.3 solution and extracted with EA. The organic
layer was dried over Na.sub.2SO.sub.4 and concentrated. The residue
was purified by column on silica gel (DCM:MeOH=100:1 to 40:1) to
give P19-1 as a white solid (759 mg, 52%).
[0309] Preparation of (P19-2): P19-1 (750 mg, 1.14 mmol) was
dissolved in saturated NH.sub.3 in MeOH solution. The mixture was
stirred for 2 hours at R.T. The solution was concentrated to
dryness to give crude P19-2 as a yellow solid (662 mg, 100%).
.sup.1H NMR (DMSO-d6, 400 MHz) .delta. 8.60 (s, 1H), 8.28 (s, 1H),
7.48 (d, J=7.6 Hz, 1H), 7.12-7.29 (m, 12H), 6.83 (d, J=8.8 Hz, 2H),
6.29 (d, J=7.6 Hz, 1H), 5.88 (d, J=8.8 Hz, 1H), 5.10 (d, J=4.8 Hz,
1H), 4.42-4.45 (m, 1H), 3.72 (s, 3H), 1.64-1.91 (m, 2H), 1.10-1.13
(m, 2H), 0.83-0.86 (m, 3H). .sup.31P NMR (CD.sub.3OD, 400 MHz)
.delta. -4.48; Negative-ESI-LCMS: m/z 606 [M-H].sup.-.
[0310] Preparation of (P19-3): P19-2 (292 mg, 0.47 mmol) was
co-evaporated with pyridine twice and dissolved in anhydrous DMF
(0.5 mL). DIPEA (1.2 mL) was added and followed by
2,2-dimethyl-propionic acid iodomethyl ester (680 mg, 2.8 mmol).
The reaction mixture was stirred at R.T. under N.sub.2 for 16
hours. The reaction was quenched by Na.sub.2S.sub.2O.sub.3 solution
and extracted with EA. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by
column on silica gel (DCM:MeOH=100:1 to 30:1) to give P19-3 as a
white solid (95 mg, 30%).
[0311] Preparation of (19a): P19-3 (95 mg, 0.13 mmol) was dissolved
in a 80% HCOOH aq. solution, and the mixture was stirred at R.T.
for 16 hours. The solvent was removed, and the residue was purified
by RP HPLC (MeCN and 0.1% HCOOH in water) to give compound 19a as a
white solid (10 mg, 17%). .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta.
7.69 (d, J=7.2 Hz, 1H), 5.91 (d, J=7.6 Hz, 1H), 5.84 (d, J=22.0 Hz,
1H), 5.73 (d, J=14.0 Hz, 2H), 5.52 (d, J=5.2 Hz, 1H), 5.13-5.22 (m,
1H), 4.53-4.61 (m, 1H), 4.31 (d, J=9.6 Hz, 1H), 1.92-2.08 (m, 2H),
1.23 (s, 9H), 1.03-1.07 (m, 3H); .sup.31P NMR (CD.sub.3OD, 162 MHz)
.delta. -7.93; ESI-LCMS: m/z 450 [M+H].sup.+.
##STR00181## ##STR00182##
[0312] Preparation of (P20-1): To a stirred suspension of P3-1
(20.0 g, 81.3 mmol), imidazole (15.9 g, 234.0 mmol), PPh.sub.3
(53.5 g, 203.3 mmol) and pyridine (90 mL) in anhydrous THF (360 mL)
was added dropwise a solution of I.sub.2 (41.3 g, 162.6 mmol) in
THF (350 mL) at 0.degree. C. After addition, the mixture was warmed
to R.T. and stirred for 14 hours. The solution was quenched with
aq. Na.sub.2S.sub.2O.sub.3 (150 mL) and extracted with EA. The
organic layer was dried over Na.sub.2SO.sub.4 and concentrated. The
residue was purified on a silica gel column (DCM:MeOH=100:1 to
10:1) to afford P20-1 as a white solid (22.1 g, 76.4%). H NMR
(CD.sub.3OD, 400 MHz) .delta. 7.70 (d, J=8.0 Hz, 1H), 5.88 (dd,
J.sub.1=1.6 Hz, J.sub.2=20.8 Hz, 1H), 5.71 (d, J=8.4 Hz, 1H), 5.24
(dd, J.sub.1=2.0 Hz, J.sub.2=5.2 Hz, 1H), 5.10 (dd, J.sub.1=2.0 Hz,
J.sub.2=5.2 Hz, 1H), 3.78-3.83 (m, 1H), 3.61-3.65 (m, 1H), 3.44
(dd, J.sub.1=J.sub.2=6.0 Hz, 1H).
[0313] Preparation of (P20-2): To a stirred solution of P20-1 (22.1
g, 62.1 mmol) in anhydrous THF (200 mL) was added dropwise DBU
(14.2 g, 93.1 mmol) in THF (50 mL) at 0.degree. C. over 10 mins.
The mixture was stirred at 60.degree. C. for 6 hours. The reaction
was quenched with aq. NaHCO.sub.3 (200 mL) and extracted with EA.
The organic layer was washed with brine and dried over
Na.sub.2SO.sub.4. The solvent was removed, and the residue was
purified on a silica gel column (MeOH:DCM=1/100 to 1/30) to afford
P20-2 as a white solid (8.7 g, 61.5%). .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 7.51 (d, J=8.0 Hz, 1H), 6.05 (dd, J.sub.1=1.2 Hz,
J.sub.2=17.2 Hz, 1H), 5.73 (d, J=8.0 Hz, 1H), 5.26 (dd, J.sub.1=1.2
Hz, J.sub.2=4.8 Hz, 1H), 5.13 (dd, J.sub.1=1.2 Hz, J.sub.2=4.8 Hz,
1H), 4.63 (dd, J=2.0 Hz, J.sub.2=3.2 Hz, 1H), 4.41 (dd,
J.sub.1=J.sub.2=2.0 Hz, 1H).
[0314] Preparation of (P20-3): To a stirred solution of P20-2 (3.2
g, 14.0 mmol) in anhydrous pyridine (10 mL) and DCM (100 mL) was
added dropwise a solution of TBSCl (4.2 g, 28.0 mmol) at 0.degree.
C. Stirring was continued at R.T. for 18 hours. The mixture was
diluted with DCM. The organic layer was washed with brine and dried
over Na.sub.2SO.sub.4. The solvent was removed, and the residue was
purified on a silica gel column (10% MeOH in DCM) to afford P20-3
as a white solid (3.4 g, 70.8%).
[0315] Preparation of (P20-4): To a stirred solution of NaHCO.sub.3
in H.sub.2O (250 mL) and acetone (200 mL) was added oxone
(30.0.times.4 g) at 0.degree. C. The mixture was warmed to R.T.,
and the distillate was collected at -78.degree. C. (120 mL) under
slightly reduced pressure to give a solution of DMDO in acetone. To
a stirred solution of P20-3 (250.0 mg, 0.7 mmol) in DCM (20 mL)
were added a DMDO (120 mL) solution at -40.degree. C. and
MgSO.sub.4. The mixture was warmed to R.T. and then stirred for 2
hours. The solution was filtrated, and the filtrate was used for
the next-step directly.
[0316] Preparation of (P20-5: To a stirred solution of P20-4 (500.0
mg, 1.4 mmol) in anhydrous DCM (50 mL) was added
allyl-trimethyl-silane (760.0 mg, 6.7 mmol) and SnCl.sub.4 (1.2 g,
4.5 mmol) at -40.degree. C. The mixture was warmed and stirred at
0.degree. C. for 1 hour. The reaction was quenched with saturated
NaHCO.sub.3 and extracted with DCM. The organic layer was dried
over Na.sub.2SO.sub.4 and concentrated. The residue was purified on
a silica gel column (20.about.50% EA in PE) to give P20-5 as a
white foam (120 mg, 41%). .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta.
8.01 (d, J=8.4 Hz, 1H), 6.12 (dd, J.sub.1=3.6 Hz, J.sub.2=15.2 Hz,
1H), 5.87-5.96 (m, 1H), 5.71 (d, J=8.4 Hz, 1H), 5.06-5.22 (m, 3H),
4.60 (dd, J.sub.1=5.6 Hz, J.sub.2=14.4 Hz, 1H), 3.72 (d, J=11.6 Hz,
1H), 3.48 (d, J=11.6 Hz, 1H), 2.62-2.67 (m, 1H), 2.23-2.29 (m, 1H);
ESI-LCMS: m/z=422 [M+Na].sup.+.
[0317] Preparation of (P20-6): To a stirred solution of P20-5
(270.0 mg, 0.7 mmol) in dry DCM were added imidazole (400.0 mg, 5.9
mmol) and TBSCl (390.0 mg, 2.6 mmol) at R.T. The mixture was
stirred at R.T. for 18 hours. The solution was diluted with EA. The
solvent was washed with brine and dried over Na.sub.2SO.sub.4. The
solvent was removed, and the residue was purified on a silica gel
column (20-40% EA in PE) to afford compound P20-6 as a white foam
(280 mg, 80.7%). ESI-LCMS: m/z 537 [M+Na].sup.+.
[0318] Preparation of (P20-7): To a stirred solution of P20-6
(280.0 mg, 0.5 mmol) in dry MeCN were added TPSCl (350.0 mg, 1.2
mmol), NEt.sub.3 (400.0 mg, 4.0 mmol) and DMAP (270.0 mg, 2.2 mmol)
at R.T. The mixture was stirred at R.T. for 18 hours. The solution
was quenched with ammonium. The organic layer was washed with brine
and dried over Na.sub.2SO.sub.4. The solvent was removed, and the
residue was purified by TLC (using EA) to afford compound P20-7 as
a white foam (240.0 mg, 85.7%). ESI-LCMS: m/z 514 [M+H].sup.+.
[0319] Preparation of (P2-8): To a stirred solution of P20-7 (270.0
mg, 0.5 mmol) in dry DCM were added AgNO.sub.3 (1.5 g, 8.8 mmol),
MMTrCl (450.0 mg, 1.5 mmol) and collidine (500.0 mg, 4.1 mmol) at
R.T. The mixture was stirred at R.T. for 18 hours. The solution was
diluted with DCM. The organic layer was washed with brine and dried
over Na.sub.2SO.sub.4. The solvent was removed, and the residue was
purified on a silica gel column (20-40% EA in PE) to afford
compound P20-8 as a white foam (300 mg, 81.6%). ESI-LCMS: m/z 786
[M+H].sup.+.
[0320] Preparation of (20a): To a stirred solution of P20-8 (170.0
mg, 0.3 mmol) in dry MeOH was added NH.sub.4F (300.0 mg, 8.1 mmol),
and the mixture was refluxed for 24 hours. The solvent was removed
under reduced pressure, and the residue was purified on a silica
gel column (2-5% MeOH in DCM) to give the crude product. The crude
product was further purified by RP HPLC (water and 0.1% HCOOH in
MeCN) to afford compound 20a as a white solid (47.0 mg, 49.8%).
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.13 (d, J=8.4 Hz, 1H),
6.12 (dd, J.sub.1=3.2 Hz, J.sub.2=12.0 Hz, 1H), 5.87-5.97 (m, 2H),
4.98-5.14 (m, 3H), 4.45 (dd, J.sub.1=5.2 Hz, J.sub.2=17.6 Hz, 1H),
3.71 (d, J=11.6 Hz, 1H), 3.54 (d, J=11.6 Hz, 1H), 2.54-2.59 (m,
1H), 2.33-2.39 (m, 1H); ESI-LCMS: m/z 286 [M+H].sup.+.
Example 21
Preparation of Compound (21a)
##STR00183##
[0322] Preparation of (P21-1): To a stirred solution of P20-8
(250.0 mg, 0.3 mmol) in MeOH was added Pd/C (500.0 mg), and the
mixture was stirred under H.sub.2 (balloon) for 18 hours at R.T.
The reaction was filtered, and the solvent removed under reduced
pressure. The residue was purified by prep. TLC (30% EtOAc in PE)
to afford P21-1 as a white foam (210.0 mg, 84.0%).
[0323] Preparation of (P21-2): To a stirred solution of P21-1
(210.0 mg, 0.3 mmol) in dry THF was added TBAF (1 mL, 1 mmol), and
the mixture was stirred at R.T. for 18 hours. The solvent was
removed under reduced pressure, and the residue was purified by
prep. TLC (30% EtOAc in PE) to give compound 21a as a white foam
(111.2 mg, 74.6%). .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 8.49 (s,
1H), 7.75 (d, J=6.8 Hz, 1H), 6.83-7.32 (m, 14H), 6.25 (d, J=7.6 Hz,
1H), 5.95 (dd, J.sub.1=4.8 Hz, J.sub.2=14.8 Hz, 1H), 5.48 (d, J=5.6
Hz, 1H), 4.86-5.15 (m, 2H), 4.15-4.21 (m, 1H), 3.72 (s, 3H),
3.38-3.49 (m, 2H), 1.24-1.58 (m, 4H), 0.84 (t, J=7.2 Hz, 3H);
ESI-MS: m/z 560 [M+H].sup.+.
[0324] Preparation of (P21): Compound P21-2 (81 mg) was dissolved
in a mixture (5 mL) of formic acid (80%) and water (20%). The
resulting solution was stirred at R.T. for 3 hours and then
concentrated. The residue was co-evaporated with methanol/toluene
three times. Chromatography on silica gel with 5-12% methanol in
DCM gave a mixture of two compounds, which was dissolved in
methanol with a drop of concentrated aqueous ammonia and
concentrated. The residue was purified on silica gel with 5-12%
methanol in DCM to give compound 21a (27 mg) as a white solid;
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.05 (d, J=7.6 Hz, 1H),
6.06 (dd, J.sub.1=2.8 Hz, J.sub.2=16 Hz, 1H), 5.87 (d, J=7.6 Hz,
1H), 5.10 (dd, J=3.2, 5.2 Hz, 0.5H), 4.96 (dd, 3.2, 5.2 Hz, 0.5H),
4.42 (dd, J=5.6, 17.2 Hz, 1H), 3.67 (dd, J=11.6, 76 Hz, 2H),
1.70-1.79 (m, 1H), 1.31-1.61 (m, m, 3H), 0.94 (t, J=6.8 Hz, 3H).
MS: m/z 417 [M+2-methylheptylamine]+.
Example 22
Preparation of Compound (22a)
##STR00184##
[0326] Preparation of (P22-1): To a solution of P20-2 (5.23 g, 23.1
mmol) in anhydrous MeOH (50 mL) was added PbCO.sub.3 (12.7 g, 46.3
mmol) at R.T. A solution of I.sub.2 (11.7 g, 46.3 mmol) in MeOH (10
mL) was then added dropwise at 0.degree. C. The reaction mixture
was stirred at R.T. for overnight. The reaction was quenched with
Na.sub.2S.sub.2O.sub.3 and dissolved in EA. The organic layer was
dried over Na.sub.2SO.sub.4 and concentrated. The residue was
purified by column (DCM/MeOH=100/1 to 20/1) to give P22-1 as a
white solid (5.6 g, 71.8%). .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 7.67 (d, J=8.0 Hz, 1H), 5.88 (dd, J.sub.1=J.sub.2=7.6 Hz,
1H), 5.73 (d, J=8.0 Hz, 1H), 5.24 (dd, J.sub.1=4.4 Hz, J.sub.2=6.4
Hz, 1H), 5.11 (dd, J.sub.1=6.4 Hz, J.sub.2=6.0 Hz, 1H); 4.65 (dd,
J.sub.1=20.0 Hz, J.sub.2=20.4 Hz, 1H), 3.67 (d, J=11.6 Hz, 1H),
3.54 (d, J=11.6 Hz, 1H), 3.43 (s, 3H).
[0327] Preparation of (P22-2): To a stirred solution of P22-1 (5.6
g, 14.5 mmol) in anhydrous pyridine (20 mL) was added dropwise BzCl
(2.9 g, 20.9 mmol) at 0.degree. C. The mixture was stirred at R.T.
for 10 hours. The reaction was quenched with H.sub.2O, and the
solution was concentrated. The residue was dissolved in EA and
washed with saturated NaHCO.sub.3. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified on a
silica gel column (20-40% EA in PE) to give P22-2 as a white foam
(4.9 g, 74.2%).
[0328] Preparation of (P22-3): P22-2 (4.9 g, 10.0 mmol), BzONa
(14.4 g, 100 mmol) and 15-crown-5 (22.0 g, 100 mmol) were suspended
in DMF (200 mL). The mixture was stirred at 60-70.degree. C. for 3
days. The precipitate was removed by filtration, and the filtrate
was diluted with EA. The solvent was washed with brine and dried
over Na.sub.2SO.sub.4. The solvent was removed, and the residue was
purified on a silica gel column (20-60% EA in PE) to afford P22-3
as a white foam (2.3 g, 47.9%).
[0329] Preparation of (P22-4): P22-3 (2.3 g, 4.8 mmol), DMAP (1.2
g, 9.6 mmol), TPSCl (2.9 g, 9.6 mmol) and Et.sub.3N (0.97 g, 9.6
mmol) were suspended in MeCN (10 mL). The mixture was stirred at
R.T. for 14 hours. NH.sub.3 in THF (saturated at 0.degree. C., 100
mL) was added to the mixture, and the mixture stirred at R.T. for 2
hours. The solvent was removed, and the residue was purified by
column (DCM/MeOH=100:1 to 50:1) to give the crude product (1.2 g).
The crude product was dissolved in pyridine, and BzCl (0.42 g, 3.0
mmol) was added. The mixture was stirred at R.T. for 16 hours and
quenched with water. The solvent was removed, and the residue was
purified on a silica gel column (PE:EA=2:1 to 1:1) to give P22-4 as
a white foam (460 mg, 31%).
[0330] Preparation of (22a): P22-4 (0.46 g, 0.8 mmol) was dissolved
in saturated methanolic ammonia (100 mL), and the mixture was
stirred at R.T. for 14 hours. The solvent was removed, and the
residue was dissolved in H.sub.2O and washed with DCM. The aqueous
phase was lyophilized and further purified by prep. HPLC (0.1%
formic acid in water/acetonitrile) to give compound 22a as a white
solid (145 mg, 78.9%). .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta.
7.88 (d, J=7.6 Hz, 1H), 6.03 (d, J=18.4 Hz, 1H), 5.87 (d, J=7.6 Hz,
1H), 4.86-5.00 (m, 1H), 4.49 (dd, J.sub.1=23.2 Hz, J.sub.2=22.8 Hz,
1H), 3.90 (d, J=12.0 Hz, 1H), 3.66 (d, J=12.0 Hz, 1H), 3.41 (s,
3H); ESI-MS: m/z 276 [M+H]*.
Example 23
Preparation of Compound (23a)
##STR00185## ##STR00186##
[0332] Preparation of (P23-2): To a solution of P23-1 (3.1 g, 4.5
mmol) in DMF (30 mL) was added anhydrous K.sub.2CO.sub.3 (1.24 g,
9.03 mmol) and PMBCl (1.40 g, 9.03 mmol). The mixture was stirred
at ambient temperature overnight. The reaction was quenched with
water and extracted with EA. The organic layer was concentrated,
and the residue was purified on a silica gel column (PE:EA=10:1 to
4:1) to give the intermediate as a white solid (2.36 g, 74.8%).
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 7.29-7.88 (m, 23H),
6.83-6.98 (m, 6H), 6.35-6.45 (m, 1H), 4.51-5.50 (m, 6H), 3.89-3.95
(m, 9H), 3.66-3.71 (m, 2H), 3.03 (d, J=11.2 Hz, 1H), 1.21 (s, 9H),
0.89 (m, 9H), 0.01-0.11 (m, 6H). The intermediate was used in the
next step.
[0333] To a stirred solution of the intermediate (11.0 g, 10.47
mmol) in anhydrous THF (100 mL) was added TBAF (8.20 g, 31.42 mmol)
at R.T., and the mixture was stirred at R.T. for 5 hours. The
solution was removed, and the residue was purified on a silica gel
column (PE:EA=5:1 to 1:1) to give a second intermediate as a white
solid (5.99 g, 82%).
[0334] To a stirred solution of the second intermediate (500 mg,
0.716 mmol) in anhydrous DMF (10 mL) was added NaH (51.5 mg, 2.14
mmol) and BnBr (365 mg, 2.14 mmol) dropwise at 0.degree. C. The
mixture was stirred at R.T. for overnight. The solution was
quenched with water and extracted with EA. The concentrated organic
phase was purified on a silica gel column (PE:EA=10:1 to 4:1) to
give a third intermediate as a white solid (496 mg, 79%).
[0335] The third intermediate (2.5 g, 2.84 mmol) was dissolved in
80% HOAc (25 mL) at R.T., and the mixture was stirred at R.T. for
overnight. The reaction was quenched with MeOH, and the solvent was
removed. The crude was purified on a silica gel column (PE:EA=5:1
to 1:1) to give P23-2 as a white solid (1.2 g, 73%).
[0336] Preparation of (P23-3): To a stirred solution of DAST (1.39
g, 8.68 mmol) in anhydrous toluene (15 mL) was added dropwise a
solution of P23-2 (1.0 g, 1.73 mmol) at -78.degree. C. The mixture
was stirred at -78.degree. C. for 30 mins. The solution was heated
to 60.degree. C. gradually and then stirred overnight. The mixture
was poured into saturated Na.sub.2CO.sub.3 solution. The
concentrated organic phase was purified on a silica gel column
(PE:EA=10:1 to 4:1) to give P23-3 as a white solid (449 mg, 45%).
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 7.87 (d, J=8.4 Hz, 1H),
7.27-7.37 (m, 12H), 6.82-6.84 (m, 2H), 6.14 (dd, J=16.8, 2.0 Hz,
1H), 5.18-5.50 (m, 4H), 4.96 (s, 2H), 4.45-4.88 (m, 7H), 3.67-3.89
(m, 5H).
[0337] Preparation of (P23-4): A mixture of P23-3 (1.20 g, 2.07
mmol) and CAN (3.41 g, 6.23 mmol) in a solution of MeCN:Water (3:1,
10 mL) was stirred at R.T. overnight. Brine (10 mL) was added, and
the mixture was extracted with EA. The combined organic extracts
were dried and evaporated under reduced pressure. The residue was
purification by chromatography on silica gel (PE:EA=10:1 to 2:1) to
give P23-4 as a yellow solid (475 mg, 49.8%).
[0338] Preparation of (P23-5): To a stirred solution of P23-4 (550
mg, 210 mmol) in anhydrous MeCN (10 mL) were added TPSCl (725 mg,
2.40 mmol), DMAP (293 mg, 2.40 mmol) and TEA (242 mg, 2.40 mmol) at
R.T., and the mixture was stirred at R.T. overnight. NH.sub.4OH (25
mL) was added, and the mixture was stirred for 2 hours. The solvent
was removed, and the residue was purified on a silica gel column
(PE:EA=8:1 to 2:1) to give P23-5 as a white solid (700 mg crude).
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 7.86 (d, J=8.4 Hz, 1H),
7.27-7.36 (m, 10H), 6.13 (dd, J.sub.1=17.2 Hz, J.sub.2=2.0 Hz, 1H),
5.48-5.53 (m, 1H), 5.11-5.26 (m, 1H), 4.44-4.74 (m, 7H), 3.89 (dd,
J.sub.1=10.4 Hz, J.sub.2=2.0 Hz, 1H), 3.69 (dd, J.sub.1=10.8 Hz,
J.sub.2=1.6 Hz, 1H).
[0339] Preparation of (P23-6): To a stirred solution of P23-5 (1.0
g, 2.18 mmol) in anhydrous DCM (15 mL) was added MMTrCl (2.02 g,
6.56 mmol) and AgNO.sub.3 (1.11 g, 6.56 mmol) at R.T., and the
mixture was stirred at R.T. overnight. The solid was filtered off
and washed with DCM. The filtrate was washed with brine and dried
over Na.sub.2SO.sub.4. The organic phase was concentrated, and the
residue was purified on a silica gel column (PE:EA=8:1 to 2:1) to
give P23-6 as a white solid (520 mg, 41%).
[0340] Preparation of (P23-7): To a stirred solution of P23-6 (520
mg, 0.713 mmol) in acetone were added ammonium formate (2.0 g, 31.7
mmol, in portions) and 10% palladium on carbon (1.0 g). The mixture
was refluxed for 12 hours. The catalyst was filtered off and washed
with solvent. The filtrate was added EA and washed with brine. The
concentrated organic phase was purified by column chromatography
(DCM:MeOH=100:1 to 15:1) and prep. TLC to give P23-7 as a white
solid (270 mg, 69.0%). .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta.
8.54 (s, 1H), 7.73 (d, J=7.6 Hz, 1H), 7.13-7.32 (m, 12H), 6.83 (d,
J=8.4 Hz, 2H), 6.29 (d, J=7.6 Hz, 1H), 5.99-6.04 (m, 1H), 5.82 (d,
J=5.6 Hz, 1H), 5.39 (t, J=5.2 Hz, 1H), 5.09 (t, J=5.2 Hz, 1H),
4.32-4.58 (m, 3H), 3.54-3.72 (m, 5H). ESI-MS: m/z 549.6 [M+H]*.
[0341] Preparation of (23a): P23-7 (130 mg, 0.236 mmol) was
dissolved in 80% HCOOH (20 mL) at R.T., and the mixture was stirred
at 50.degree. C. for 12 hours. The solvent was removed, and the
residue was co-evaporated with toluene twice. The residue was
re-dissolved in MeOH (20 mL) at 60.degree. C. and stirring was
continued for 48 hours. The solvent was removed, and the residue
was purified by column chromatography (DCM:MeOH=100:1 to 10:1) to
give compound 23a as a white solid (45 mg, 69.0%). .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.00 (d, J=7.6 Hz, 1H), 6.13 (dd,
J.sub.1=16.0 Hz, J.sub.2=4.0 Hz, 1H), 5.89 (d, J=7.6 Hz, 1H),
5.18-5.21 (m, 1H), 5.05-5.07 (m, 1H), 4.60 (s, 1H), 4.51-4.57 (m,
2H), 3.84 (dd, J=12.0 Hz, J.sub.2=2.0 Hz, 1H), 3.75 (dd,
J.sub.1=12.0 Hz, J.sub.2=2.0 Hz, 1H). ESI-MS: m/z 277.8
[M+H].sup.+, 554.8 [2M+H].sup.+.
Example 24
Preparation of Compound (24a)
##STR00187## ##STR00188## ##STR00189##
[0343] Preparation of (P24-2): To a solution of P24-1 (30.0 g,
100.0 mmol) in pyridine (300 mL) was added BzCl (56.0 g, 400 mmol)
at 25.degree. C. The mixture was stirred at 25.degree. C. for 15
hours. The mixture was concentrated and purified by column
chromatography (PE:EA=20:1 to 2:1) to give crude P24-2 (55.0 g,
81%).
[0344] Preparation of (P24-3): P24-2 (55.0 g, 92 mmol) was
dissolved in 80% HOAc aq. solution, and the mixture was refluxed
for 14 hours. The solvent was removed under reduced pressure, and
the residue was co-evaporated with toluene. The residue was
purified on a silica gel column (PE/EA=4:1 to 2:1) to give P24-3 as
a white solid (39.2 g, 83%).
[0345] Preparation of (P24-4): P24-3 (39.2 g, 83 mmol) was
dissolved in saturated methanolic ammonia, and the resulting
solution was stirred at R.T. for 15 hours. The solvent was removed,
and the residue was purified on a silica gel column (DCM/MeOH=50:1
to 20:1) to give P24-4 (21.0 g, 95.8%).
[0346] Preparation of (P24-5): To a solution of P24-4 (21.0 g, 79.5
mmol) in pyridine (250 mL) was added DMTrCl (28.2 g, 83.5 mmol) at
0.degree. C. The solution was stirred at R.T. for 15 hours. The
reaction was quenched with MeOH and concentrated to dryness under
reduced pressure. The residue was dissolved in EtOAc and washed
with water. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated. The residue was dissolved in DCM (300 mL). Imidazole
(13.6 g, 200 mmol) and TBSCl (30.0 g, 200 mmol) were added. The
reaction mixture was stirred at R.T. for 12 hours. The reaction
mixture was washed with NaHCO.sub.3 and brine. The organic layer
was dried over Na.sub.2SO.sub.4 and concentrated. The residue (48.5
g, 79.5 mmol) was dissolved in 80% HOAc aq. solution (400 mL). The
mixture was stirred at R.T. for 20 hours. The mixture was diluted
with EtOAc and washed with NaHCO.sub.3 solution and brine. The
organic layer was dried over Na.sub.2SO.sub.4 and purified by
silica gel column chromatography (1-2% MeOH in DCM) to give P24-5
as a white solid (21.0 g, 70%). .sup.1H NMR (400 MHz, MeOD) .delta.
7.83 (d, J=8.0 Hz, 1H), 6.14 (dd, J.sub.1=6.0 Hz, J.sub.2=10.0 Hz,
1H), 5.73 (d, J=8.4 Hz, 1H), 4.38-4.46 (m, 1H), 3.89-3.91 (m, 1H),
3.88 (dd, J.sub.1=2.8 Hz, J.sub.2=5.2 Hz, 1H), 3.72 (dd, J=2.8 Hz,
J.sub.2=5.2 Hz, 1H), 0.93 (s, 9H), 0.15 (m, 6H). ESI-MS: m/z 379.1
[M+H].sup.+.
[0347] Preparation of (P24-6): To a solution of P24-5 (21.0 g, 55.6
mmol) in anhydrous CH.sub.3CN (200 mL) was added IBX (17.1 g, 61.1
mmol) at R.T. The reaction mixture was refluxed for 1 hour and then
cooled to 0.degree. C. The precipitate was filtered off, and the
filtrate was concentrated to give the aldehyde as a yellow solid
(21.0 g, 55.6 mmol). To a solution of the aldehyde (21.0 g, 55.6
mmol) in dioxane (200 mL) were added 37% CH.sub.2O (22.2 mL, 222.4
mmol) and 2N NaOH aq. solution (55.6 mL, 111.2 mmol). The mixture
was stirred at R.T. for 2 hours and then neutralized with AcOH to
pH=7. To the reaction were added EtOH (50 mL) and NaBH.sub.4 (12.7
g, 333.6 mmol). The mixture was stirred at R.T. for 30 mins. The
reaction was quenched with saturated aq. NH.sub.4Cl. extracted with
EA. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by silica gel column
chromatography (1-3% MeOH in DCM) to give P24-6 as a white solid
(13.5 g, 59.5%).
[0348] Preparation of (P24-7): To a solution of P24-6 (13.5 g, 33.1
mmol) in DCM (100 mL) were added pyridine (20 mL) and DMTrCl (11.2
g, 33.1 mmol) at 0.degree. C. The solution was stirred at
25.degree. C. for 3 hours, and then treated with MeOH (30 mL). The
solvent was removed, and the residue was purified by silica gel
column chromatography (DCM:MeOH=300:1 to 100:1) to give a residue.
The residue was dissolved in anhydrous pyridine (150 mL) and
TBDPSCl (16.5 g, 60 mmol) and AgNO.sub.3 (10.2 g, 60 mmol) were
added. The mixture was stirred at 25.degree. C. for 15 hours, and
then filtered and concentrated. The mixture was dissolved in EtOAc
and washed with brine. The organic layer was dried over
Na.sub.2SO.sub.4. Purified by silica gel column chromatography
(DCM:MeOH=300:1 to 100:1) gave the product as a yellow solid (16.2
g, 85.3%). The solid was dissolved in 80% HOAc aq. solution (400
mL). The mixture was stirred at R.T. for 15 hours. The mixture was
diluted with EtOAc and washed with NaHCO.sub.3 solution and brine.
The organic layer was dried over Na.sub.2SO.sub.4 and purified by
silica gel column chromatography (DCM:MeOH=200:1 to 50:1) to give
P24-7 as a white solid (9.5 g, 86.5%). .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 7.39-7.70 (m, 11H), 6.34-6.38 (m, 1H), 5.12 (d, J=8.0
Hz, 1H), 4.79 (dd, J.sub.1=10.0 Hz, J.sub.2=16.0 Hz, 1H), 4.14 (dd,
J.sub.1=1.6 Hz, J.sub.2=11.6 Hz, 1H), 3.48-3.84 (m, 2H), 3.49 (dd,
J.sub.1=1.6 Hz, J.sub.2=11.6 Hz, 1H), 1.12 (s, 9H), 0.92 (s, 9H),
0.16 (s, 6H).
[0349] Preparation of (P24-8): To a solution of P24-7 (6.0 g, 9.3
mmol) in anhydrous DCM (80 mL) was added Dess-Martin periodinane
(7.9 g, 18.6 mmol) at 0.degree. C. under nitrogen. The reaction was
stirred at R.T. for 1 hour. The solvent was removed in vacuo, and
the residue was triturated with diethyl ether (50 mL). The mixture
was filtered through a pad of MgSO.sub.4, and the organic solvent
was stirred with an equal volume of
Na.sub.2S.sub.2O.sub.3.5H.sub.2O in saturated NaHCO.sub.3 (50 mL)
until the organic layer became clear (approx. 10 min). The organic
layer was separated, washed with brine, and dried over MgSO.sub.4.
After concentration in vacuo, P24-8 was obtained as a red solid
(5.8 g. 98%).
[0350] Preparation of (P24-9): To a mixture of
methyltriphenylphosphonium bromide (9.6 g, 27.0 mmol) in anhydrous
THF (60 mL) was added n-BuLi (10.8 mL, 27.0 mmol) at -70.degree. C.
under nitrogen. The reaction was stirred at 0.degree. C. for 30
mins. A solution of P24-8 (5.8 g, 9.0 mmol) in anhydrous THF (20
mL) was added dropwise at 0.degree. C. under nitrogen. The reaction
was stirred at R.T. for 12 hours. The reaction was quenched with
NH.sub.4Cl and extracted with EtOAc. The organic layer was
separated, dried and concentrated, and the residue was purified by
silica gel column chromatography (DCM:MeOH=300:1 to 100:1) to give
P24-9 as a white solid (3.0 g, 51%).
[0351] Preparation of (P24-10): To a solution of P24-9 (2.9 g, 4.5
mmol) in anhydrous MeOH (20 mL) was added Pd/C (1.4 g) at
25.degree. C. under hydrogen atmosphere. The mixture was stirred at
25.degree. C. for 1 hour. The solution was filtered, evaporated to
dryness and purified on a silica gel column (DCM:MeOH=3 00:1 to
100:1) to give P24-10 as a white solid (2.3 g, 79.3%).
[0352] Preparation of (P24-11): To a solution of P24-10 (1.0 g,
1.55 mmol) in anhydrous CH.sub.3CN (20 mL) were added TPSCl (940
mg, 3.1 mmol), DMAP (380 mg, 3.1 mmol) and NEt.sub.3 (470 mg, 4.6
mmol) at R.T. The reaction was stirred at R.T. for 5 hours.
NH.sub.4OH (8 mL) was added, and the reaction was stirred for 1
hour. The mixture was diluted with DCM (150 mL) and washed with
water, 0.1 M HCl and saturated aq. NaHCO.sub.3. The solvent was
removed, and the residue was purified by silica gel column
chromatography (PE:EA=10:1 to 1:1) to give the crude product as a
yellow solid (900 mg, 90%). To a solution of the crude product in
DCM (10 mL) were added MMTrCl (930 mg, 3.0 mmol), AgNO.sub.3 (510
mg, 3.0 mmol) and colliding (720 mg, 6.0 mmol) at R.T. The reaction
was stirred for 12 hours at R.T. The reaction was filtered,
concentrated and purified by silica gel column chromatography
(DCM:MeOH=200:1 to 50:1) to give P24-11 as a yellow solid (1.1 g,
77.6%).
[0353] Preparation of (P24-12): To a solution of P24-11 (1.1 g, 1.2
mmol) in MeOH (40 mL) was added NH.sub.4F (1.0 g, 30 mmol) at
25.degree. C. and stirred at 70.degree. C. for 15 hours. The
solution was filtered and evaporated to dryness, and the residue
was purified by silica gel column (DCM:MeOH=200:1 to 20:1) to give
P24-12 as a white solid (450 mg, 66.6%). H NMR (400 MHz, MeOD)
.delta. 8.58 (s, 1H), 7.62 (d, J=7.6 Hz, 1H), 7.13-7.30 (m, 12H),
6.83-6.85 (m, 2H), 6.29 (d, J=7.6 Hz, 1H), 6.18 (d, J=6.0 Hz, 1H),
5.94 (t, J=8.0 Hz, 1H), 5.22 (t, J=5.2 Hz, 1H), 4.28-4.37 (m, 1H),
3.72 (s, 3H), 3.57-3.62 (m, 1H), 1.39-1.60 (m, 2H), 0.79-0.84 (m,
3H). ESI-LCMS: m/z 563.6 [M+H].sup.+.
[0354] Preparation of (24a): P24-12 (250 mg, 0.44 mmol) was
dissolved in 80% HCOOH in H.sub.2O (6.0 g) at 25.degree. C. The
mixture was stirred at 35.degree. C. for 15 hours. The solution was
evaporated to dryness, dissolved in MeOH (30 mL) and stirred at
60.degree. C. for 12 hours. The solution was evaporated to dryness
and purified by silica gel column chromatography (DCM:MeOH=100:1 to
100:1) to give compound 24a as a white solid (125.6 mg, 97%).
.sup.1H NMR (400 MHz, MeOD) .delta. 7.91 (d, J=7.6 Hz, 1H), 6.19
(t, J=7.6 Hz, 1H), 5.90 (d, J=7.2 Hz, 1H), 4.47 (t, J=13.6 Hz, 1H),
3.67 (d, J=12.0 Hz, 1H), 3.52 (d, J=12.0 Hz, 1H), 1.73-1.82 (m,
1H), 1.53-1.63 (m, 1H), 095 (t, J=7.6 Hz, 3H). ESI-LCMS: m/z 291.9
[M+H].sup.+.
Example 25
Preparation of Compound (25a)
##STR00190## ##STR00191##
[0356] Preparation of (P25-2): To a solution of P25-1 (20.0 g,
70.16 mmol) in anhydrous pyridine (200 mL) was added imidazole
(19.08 g, 280.7 mmol) and TBSCl (42.10 g, 280.7 mmol) at 25.degree.
C. The solution was stirred at 25.degree. C. for 15 hours, and then
concentrated to dryness under reduced pressure. The residue was
washed with EtOAc to give the crude product as a white solid (36.4
g). The crude product was dissolved in THF (150 mL) and H.sub.2O
(100 mL), and then HOAc (300 mL) was added. The solution was
stirred at 80.degree. C. for 13 hours. The reaction was cooled to
R.T., and the mixture was concentrated to dryness under reduced
pressure. The residue was dissolved washed with EtOAc and dried to
give P25-2 as a white solid (31.2 g, 60.9%).
[0357] Preparation of (P25-3): To a stirred solution of P25-2 (31.2
g, 78.2 mmol) in anhydrous pyridine (300 mL) was added Ac.sub.2O
(11.96 g, 117.3 mmol). The mixture was stirred at 25.degree. C. for
18 hours. MMTrC (72.3 g, 234.6 mmol) and AgNO.sub.3 (39.9 g, 234.6
mmol) were then added. The solution was stirred at 25.degree. C.
for 15 hours. And H.sub.2O was added to quench the reaction. The
solution was concentrated to dryness under reduced pressure. The
residue was dissolved in EtOAc and washed with water. The organic
layer was dried over Na.sub.2SO.sub.4 and filtered. The filtrate
was concentrated in vacuo to give a residue. The residue was
purified by silica gel (DCM:MeOH=200:1 to 50:1) to give the
product. The product was dissolved in NH3/MeOH (300 mL), and the
mixture was stirred at 25.degree. C. for 20 hours. The solvent was
removed, and the residue was purified on a silica gel column
(DCM:MeOH=100:1 to 50:1) to give P25-3 as a yellow solid (28.6 g,
86.5%). .sup.1H NMR (400 MHz, MeOD) .delta. 8.01 (s, 1H), 7.23-7.35
(m, 12H), 6.85-6.87 (m, 2H), 5.60 (dd, J.sub.1=11.2 Hz, J.sub.2=5.6
Hz, 1H), 4.78-4.94 (m, 1H), 4.44 (dd, J.sub.1=8.0 Hz, J.sub.2=4.8
Hz, 1H), 3.78 (s, 3H), 3.60-3.63 (m, 1H), 3.50 (dd, J.sub.1=32.0
Hz, J.sub.2=12.0 Hz, 2H), 3.32 (s, 3H), 0.94 (s, 9H), 0.12-0.14 (m,
6H).
[0358] Preparation of (P25-4): To a solution of P25-3 (7.24 g,
10.79 mmol) in anhydrous CH.sub.3CN (100 mL) was added IBX (3.93 g,
14.03 mmol) at 20.degree. C. The reaction mixture was refluxed at
90.degree. C. for 1 hour. The reaction was filtered, and the
filtrate was concentrated to give the aldehyde as a yellow solid
(7.1 g). To a solution of the aldehyde (7.1 g, 10.6 mmol) in
dioxane (80 mL) was added 37% CH.sub.2O (4.2 mL, 42.4 mmol) and 2N
NaOH aq. solution (8.0 mL, 15.9 mmol). The mixture was stirred at
25.degree. C. for 2 hours and then neutralized with AcOH to pH=7.
To reaction was added EtOH (30 mL) and NaBH.sub.4 (2.4 g, 63.6
mmol), the reaction was then stirred for 30 mins. The mixture was
quenched with saturated aq. NH.sub.4Cl. The mixture was extracted
with EA, and the organic layer was dried over Na.sub.2SO.sub.4. The
solvent was removed, and the residue was purified by silica gel
column chromatography (DCM:MeOH=200:1 to 50:1) to give P25-4 as a
yellow solid (4.86 g, 65.4%).
[0359] Preparation of (P25-5): To a solution of P25-4 (3.8 g, 5.4
mmol) in DCM (40 mL) were added pyridine (10 mL) and DMTrCl (1.8 g,
5.4 mmol) at 0.degree. C. The solution was stirred at 25.degree. C.
for 1 hour. The reaction mixture was treated with MeOH (15 mL) and
concentrated. The residue was purified by silica gel column
chromatography (DCM:MeOH=200:1 to 50:1) to give the mono-DMTr
protected intermediate as a yellow solid (3.6 g, 66.4%). To a
solution of the intermediate in anhydrous pyridine (30 mL) were
added TBDPSCl (2.96 g, 10.8 mmol) and AgNO.sub.3 (1.84 g, 10.8
mmol). The mixture was stirred at 25.degree. C. for 15 hours. The
mixture was filtered and concentrated, and then dissolved in EtOAc
and washed with brine. The organic layer was dried over
Na.sub.2SO.sub.4, and then concentrated. The residue was purified
by silica gel column chromatography (DCM:MeOH=200:1 to 50:1) to
give the pure intermediate as a white solid (3.8 g, 85.1%). To a
solution of the intermediate (3.6 g, 2.9 mmol) in anhydrous DCM (50
mL) was added Cl.sub.2CHCOOH (1.8 mL) in anhydrous DCM (18 mL) at
-78.degree. C. The mixture was stirred at -10.degree. C. for 30
mins. The mixture was quenched with saturated aq. NaHCO.sub.3 and
extracted with DCM. The organic layer was dried over
Na.sub.2SO.sub.4, and then purified by silica gel column
chromatography (DCM:MeOH=200:1 to 50:1) to give P25-5 as a white
solid (2.2 g, 80.7%).
[0360] Preparation of (P25-6): P25-5 (2.2 g, 2.3 mol) was added to
a suspension of Dess-Martin periodinane (2.5 g, 5.8 mol) in
anhydrous CH.sub.2Cl.sub.2 (30 mL) at 25.degree. C. The mixture was
stirred at 25.degree. C. for 4 hours. The solvent was removed in
vacuo, and the residue triturated with diethyl ether (30 mL). The
mixture was filtered through a pad of MgSO.sub.4. The organic
solvent was stirred with an equal volume of
Na.sub.2S.sub.2O.sub.3.5H.sub.2O in saturated NaHCO.sub.3 (30 mL)
until the organic layer became clear (approx. 10 min). The organic
layer was separated, washed with brine, and dried over MgSO.sub.4.
The solvent was removed in vacuo to give P25-6 as a yellow solid
(2.1 g, 95%).
[0361] Preparation of (P25-7): To a stirred solution of
methyl-triphenyl-phosphonium bromide (2.3 g, 6.6 mmol) in anhydrous
THF (30 mL) was added dropwise n-BuLi (2.6 mL, 6.6 mmol, 2.5 M in
THF) at -78.degree. C. over 1 minute. Stirring was continued at
0.degree. C. for 1 hour. P25-6 (2.1 g, 2.2 mmol) was added to the
mixture, and then stirred at 25.degree. C. for 15 hours. The
reaction was quenched with saturated NH.sub.4Cl (50 mL). The
mixture was extracted with EtOAc. The combined organic phase was
dried with Na.sub.2SO.sub.4, filtered and evaporated to dryness to
give a light yellow oil. The oil was purified by column
chromatography (DCM:MeOH=200:1 to 50:1) to give P25-7 as a white
solid (1.6 g, 76%).
[0362] Preparation of (P25-8): To a solution of P25-7 (1.6 g, 1.7
mmol) in MeOH (50 mL) was added NH.sub.4F (1.5 g, 40 mmol), and the
mixture was stirred at 70.degree. C. for 15 hours. The solution was
filtered and evaporated to dryness. The residue was purified by
silica gel column (DCM:MeOH=200:1 to 20:1) to give P25-8 as a white
solid (450 mg, 49%). .sup.1H NMR (400 MHz, MeOD) .delta. 7.95 (s,
1H), 7.21-7.33 (m, 12H), 6.82-6.84 (m, 2H), 5.92 (dd, J.sub.1=11.2
Hz, J.sub.2=17.6 Hz, 1H), 5.55-5.59 (m, 1H), 5.18-5.31 (m, 2H),
4.54-4.68 (m, 1H), 4.26-4.33 (m, 1H), 3.76 (s, 3H), 3.43 (dd,
J.sub.1=12.4 Hz, J.sub.2=36.4 Hz, 2H). ESI-LCMS: m/z 584.1
[M+H].sup.+.
[0363] Preparation of (25a): P25-8 (130 mg, 0.22 mmol) was
dissolved in 80% HCOOH and the mixture was stirred at 25.degree. C.
for 1 hour. Then the solution was evaporated to dryness. The
residue was dissolved in MeOH (30 mL) and stirred at 60.degree. C.
for 12 hours. Then the solution was evaporated to dryness, and the
residue was washed by EtOAc to give P25 as a white solid (52.3 mg,
76%). .sup.1H NMR (400 MHz, MeOD) .delta. 8.03 (s, 1H), 6.17 (dd,
J.sub.1=3.2 Hz, J.sub.2=16.8 Hz, 1H), 6.03 (dd, J.sub.1=11.2 Hz,
J.sub.2=17.2 Hz, 1H), 5.50 (dd, J=1.6 Hz, J.sub.2=17.2 Hz, 1H),
5.23-5.38 (m, 2H), 4.76 (dd, J.sub.1=4.8 Hz, J.sub.2=18.0 Hz, 1H),
3.60 (dd, J.sub.1=12.0 Hz, J.sub.2=44.8 Hz, 2H). ESI-MS: m/z 334.1
[M+Na].sup.+.
Example 26
Preparation of Compound (26a)
##STR00192##
[0365] Preparation of (P26-1): To a stirred solution of P25-6 (2.1
g, 2.2 mmol) in pyridine was added HONH.sub.2.HCl (0.61 g, 8.8
mmol) at 25.degree. C. The mixture was stirred at 25.degree. C. for
2 hours. The mixture was concentrated, and the residue was purified
by column chromatography (DCM:MeOH=200:1 to 50:1) to give P26-1 as
a white solid (1.8 g, 83%).
[0366] Preparation of (P26-2): To a stirred solution of P26-1 (1.4
g, 1.47 mmol) in DCM were added TEA (0.44 g, 4.4 mmol) and
methanesulfonyl chloride (0.34 g, 2.9 mmol) at 0.degree. C. The
mixture was stirred at 25.degree. C. for 1 hour. The mixture was
quenched with saturated aq. NaHCO.sub.3 and extracted with DCM. The
organic phase was dried with Na.sub.2SO.sub.4, filtered and
evaporated. The residue was purified by column chromatography
(DCM:MeOH=200:1 to 50:1) to give P26-2 as a white solid (1.1 g,
79%).
[0367] Preparation of (P26-3): To a solution of P26-2 (1.1 g, 1.18
mmol) in MeOH (50 mL) was added NH.sub.4F (1.5 g, 40 mmol), and the
mixture was stirred at 70.degree. C. for 15 hours. The solution was
filtered and evaporated to dryness. The residue was purified by
silica gel column (DCM:MeOH=200:1 to 20:1) to give P26-3 as a white
solid (400 mg, 71%). .sup.1H NMR (400 MHz, MeOD) .delta. 7.80 (s,
1H), 7.20-7.32 (m, 12H), 6.86-6.88 (m, 2H), 5.82 (dd, J.sub.1=2.0
Hz, J.sub.2=20.0 Hz, 1H), 4.51-4.66 (m, 1H), 3.94 (dd, J.sub.1=5.2
Hz, J.sub.2=20.8 Hz, 1H), 3.78 (s, 3H), 3.56 (dd, J.sub.1=12.4 Hz,
J.sub.2=42.0 Hz, 2H). ESI-LCMS: m/z 583.1 [M+H].sup.+.
[0368] Preparation of (26a): P26-3 (200 mg, 0.34 mmol) was
dissolved in 80% HCOOH aq. solution. The mixture was stirred at
25.degree. C. for 1 hour. The solution was evaporated to dryness,
dissolved in MeOH (30 mL) and stirred at 60.degree. C. for 12
hours. The solvent was removed, and the residue was washed by EtOAc
to give compound 26a as a white solid (100.4 mg, 95%). .sup.1H NMR
(400 MHz, MeOD) .delta. 7.90 (s, 1H), 6.34 (dd, J.sub.1=2.0 Hz,
J.sub.2=19.6 Hz, 1H), 5.49 (ddd, J.sub.1=1.6 Hz, J.sub.2=4.4 Hz,
J=52.4 Hz, 1H), 5.01 (dd, J.sub.1=4.8 Hz, J.sub.2=20.8 Hz, 1H),
3.93 (dd, J.sub.1=12.4 Hz, J.sub.2=44.8 Hz, 2H). ESI-MS: m/z 311.1
[M+H].sup.+.
Example 27
Preparation of Compound (27a)
##STR00193##
[0370] Preparation of (P27-1): To a stirred solution of
chloromethyl-triphenyl-phosphonium chloride (1.9 g, 5.4 mmol) in
anhydrous THF (30 mL) was added dropwise n-BuLi (2.16 mL, 5.4 mmol,
2.5 M in THF) at -78.degree. C. over 10 mins. Stirring was
continued at -78.degree. C. for 2 hours. P25-6 (1.7 g, 1.8 mmol)
was added, and the mixture and stirred at 25.degree. C. for 15
hours. The reaction was quenched with saturated NH.sub.4Cl (50 mL).
The mixture was extracted with EtOAc. The combined organic phase
was dried with Na.sub.2SO.sub.4, filtered and evaporated to dryness
to give a light yellow oil. The oil was purified by column
chromatography (DCM:MeOH=200:1 to 50:1) to give P27-1 as a white
solid (1.2 g, 70%).
[0371] Preparation of (P27-2): To a stirred solution of P27-1 (1.2
g, 1.3 mmol) in anhydrous THF (20 mL) was added dropwise n-BuLi
(8.0 mL, 20 mmol, 2.5 M in THF) at -78.degree. C. over 10 minutes.
Stirring was continued at -78.degree. C. for 4 hours. The reaction
was quenched with saturated NH.sub.4Cl (50 mL). The mixture was
extracted with EtOAc (50.times.2 mL). The combined organic phase
was dried over Na.sub.2SO.sub.4, filtered and evaporated to
dryness. The residue was purified by column chromatography
(DCM:MeOH=200:1 to 50:1) to give P27-2 as a white solid (1.0 g,
83%).
[0372] Preparation of (P27-3): To a solution of P27-2 (1.0 g, 1.1
mmol) in MeOH (40 mL) was added NH.sub.4F (1.5 g, 40 mmol), and the
mixture was stirred at 70.degree. C. for 25 hours. The solution was
filtered, and the filtrate was evaporated to dryness. The residue
was purified on a silica gel column (DCM:MeOH=200:1 to 20:1) to
give P27-3 as a white solid (240 mg, 38%). .sup.1H NMR (400 MHz,
MeOD) .delta. 7.85 (s, 1H), 7.21-7.31 (m, 12H), 6.84-6.87 (m, 2H),
5.67 (dd, J=1.6 Hz, J.sub.2=19.2 Hz, 1H), 4.47-4.62 (m, 1H), 3.94
(dd, J.sub.1=5.2 Hz, J.sub.2=22.4 Hz, 1H), 3.77 (s, 3H), 3.56 (dd,
J.sub.1=12.4 Hz, J.sub.2=47.2 Hz, 2H), 3.04 (s, 1H). ESI-LCMS: m/z
582.1 [M+H].sup.+.
[0373] Preparation of (27a): P27-3 (130 mg, 0.22 mmol) was
dissolved in 80% HCOOH aq. solution. The mixture was stirred at
25.degree. C. for 1 hour. The solution was evaporated to dryness.
The residue was dissolved in MeOH (30 mL) and stirred at 60.degree.
C. for 12 hours. The solvent was removed, and the residue was
washed with EtOAc to give compound 27a as a white solid (43.0 mg,
63%). .sup.1H NMR (400 MHz, MeOD) .delta. 7.95 (s, 1H), 6.22 (dd,
J.sub.1=2.4 Hz, J.sub.2=18.4 Hz, 1H), 5.49 (ddd, J.sub.1=2.0 Hz,
J.sub.2=4.8 Hz, J.sub.3=53.2 Hz, 1H), 4.77 (dd, J.sub.1=5.2 Hz,
J.sub.2=20.0 Hz, 1H), 3.79 (dd, J.sub.1=12.4 Hz, J.sub.2=46.8 Hz,
2H), 3.12 (s, 3H). ESI-MS: m/z 310.1 [M+H].sup.+.
Example 28
Preparation of Compound (28a)
##STR00194##
[0375] Preparation of (P28-1): To a stirred solution of P25-1 (5.7
g. 20 mmol) in anhydrous pyridine (20 mL) was added dropwise
Ac.sub.2O (5.8 mL, 60 mmol) at 0.degree. C. The mixture was stirred
at R.T. for 10 hours. AgNO3 (8.5 g, 50 mmol) and MMTrCl (15.5 g, 50
mmol) were added. The mixture was stirred at R.T. for 10 hours. The
solution was quenched with saturated NaHCO.sub.3 and extracted with
EA. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified on a silica gel column
(DCM/MeOH=100:1 to 50:1) to afford the intermediate as a light
yellow solid (12.1 g, 93.4%). The solid was treated with saturated
NH.sub.3 in MeOH at R.T. for 14 hours. The solvent was removed, and
the residue was purified by silica gel column chromatography
(DCM/MeOH=80:1 to 30:1) to afford P28-1 as a white solid (9.2 g,
87.5%).
[0376] Preparation of (P28-2): To a stirred solution of P28-1 (9.2
g, 16.5 mmol) in dry THF (300 mL) were added imidazole (9.0 g, 132
mmol) and PPh.sub.3 (34.8 g, 132 mmol). A solution of I.sub.2 (26.0
g, 103 mmol) in THF (100 mL) was added dropwise under N.sub.2 at
0.degree. C. The mixture was stirred at R.T. for 18 hours. The
reaction was quenched with Na.sub.2S.sub.2O.sub.3 solution, and the
mixture was extracted with EtOAc. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by
silica gel column chromatography (DCM/MeOH=80:1 to 30:1) to give
P28-2 as a light yellow solid (10.3 g, 93.4%).
[0377] Preparation of (P28-3): To a stirred solution of P28-2 (10.2
g, 15.3 mmol) in dry THF (300 mL) was added DBU (4.7 g, 30.1 mmol).
The mixture was stirred at 60.degree. C. for 8 hours. The solution
was diluted with NaHCO.sub.3 solution and extracted with EtOAc. The
organic layer was dried over Na.sub.2SO.sub.4 and concentrated. The
residue was purified by silica gel column chromatography
(PE/EtOAc=3:1 to 1:3) to afford P28-3 as a light yellow foam (6.2
g, 75.6%). .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 7.71 (s, 1H),
7.23-7.76 (m, 14H), 6.74 (d, J=0.8 Hz, 2H), 5.83-5.88 (dd,
J.sub.1=2.8 Hz, J.sub.2=16.0 Hz, 2H), 4.57-4.89 (m, 2H), 4.30-4.35
(m, 1H), 4.79 (s, 3H). ESI-MS: m/z 540 [M+H].sup.+.
[0378] Preparation of (P28-4): To a stirred solution of P28-4 (5.42
g, 10 mmol) in anhydrous CH.sub.3H (100 mL) were added PbCO.sub.3
(13.7 g, 53.1 mmol) followed by a solution of I.sub.2 (12.3 g, 48.9
mmol) in CH.sub.3OH (300 mL) at 0.degree. C. The mixture was
stirred at R.T. for 10 hours. The solution was quenched with a
Na.sub.2S.sub.2O.sub.3 solution and extracted with DCM. The organic
layer was washed with NaHCO.sub.3 solution, dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by
pre-HPLC (MeCN and 0.1% HCOOH in water) to give the pure product as
a white foam (2.4 g, 34%). The product was dissolved in dry
pyridine (20 mL) and BzCl (723 mg, 5.2 mmol) was added dropwise at
0.degree. C. The mixture was stirred at 0.degree. C. for 1 hour.
The solution was quenched with NaHCO.sub.3 solution, and extracted
with EtOAc. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by silica gel column
chromatography (PE/EtOAc=5:1 to :1) to afford P28-4 as a white
solid (2.1 g, 77.1%).
[0379] Preparation of (P28-5): P28-4 (2.0 g, 2.5 mmol), BzONa (3.6
g, 25 mmol) and 15-crown-5 (5.5 g, 25 mmol) were suspended in DMF
(50 mL). The mixture was stirred at 110-125.degree. C. for 5 days.
The precipitate was removed by filtration, and the filtrate was
diluted with EA. The solution was washed with brine and dried over
Na.sub.2SO.sub.4. The solvent was removed, and the residue was
purified on a silica gel column (PE/EA=10/1 to 2/1) to afford crude
P28-5 as a light yellow foam (1.6 g, 80%).
[0380] Preparation of (P28-6): P28-5 (1.6 g, 2.0 mmol) was
dissolved in methanolic ammonia (100 mL, saturated), and the
mixture was stirred at R.T. for 20 hours. The solvent was removed,
and the residue was purified on a silica gel column (DCM/MeOH=100:1
to 20:1) to give P28-6 as a white solid (410 mg, 34.9%). .sup.1H
NMR (400 MHz, MeOD) .delta. 7.84 (s, 1H), 7.20-7.33 (m, 12H),
6.83-6.86 (m, 2H), 5.64 (dd, J=1.6 Hz, J.sub.2=18.4 Hz, 1H),
4.46-4.62 (m, 1H), 4.08 (dd, J.sub.1=6.0 Hz, J.sub.2=22.0 Hz, 1H),
3.76 (s, 3H), 3.58 (dd, J=12.4 Hz, J.sub.2=30.4 Hz, 2H), 3.31 (s,
3H). ESI-LCMS: m/z 588.1 [M+H].sup.+.
[0381] Preparation of (28a): P28-8 (200 mg, 0.34 mmol) was
dissolved in 80% HCOOH and the mixture was stirred at 25.degree. C.
for 1 hour. The solution was evaporated to dryness, and the residue
was dissolved in MeOH (30 mL) and stirred at 60.degree. C. for 12
hours. The solvent was removed, and the residue washed with EtOAc
to give compound 28a as a white solid (46.1 mg, 43%). .sup.1H NMR
(400 MHz, MeOD) .delta.7.92 (s, 1H), 6.22 (dd, J.sub.1=1.6 Hz,
J.sub.2=18.8 Hz, 1H), 5.25 (ddd, J.sub.1=1.6 Hz, J.sub.2=6.0 Hz,
J=54.0 Hz, 1H), 4.89-4.91 (m, 1H), 3.87 (d, J=11.6 Hz, 1H), 3.67
(d, J=12.0 Hz, 1H), 3.44 (s, 3H). ESI-MS: m/z 316.1
[M+H].sup.+.
Example 29
Preparation of Compound (29a)
##STR00195##
[0383] DEAD (40% in toluene, 0.15 mL, 0.33 mmol) was added to a
stirred solution of triphenylphosphine (78 mg, 0.3 mmol) in
anhydrous 1,4-dioxane (0.5 mL) at 0.degree. C. under argon. The
mixture was warmed up to R.T. and compound 10a (26 mg, 0.1 mmol)
and bis(pivaloyloxymethyl)phosphate (98 mg, 0.3 mmol) were added.
The resulting mixture was stirred at 65.degree. C. for 3 days.
Diisopropylethylamine (50 .mu.L) was added, and the mixture was
stirred at 70.degree. C. for 3 days. Another reaction of the same
scale was conducted separately. The two reaction mixtures were
combined and concentrated. Chromatography on silica gel with 5-10%
methanol in DCM gave the desired product (20 mg) with a minor
impurity. A second chromatography on silica gel, followed by RP
HPLC with acetonitrile/water, gave the compound (2.8 mg) as a
colorless residue; .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 7.65
(d, J=8.0 Hz, 1H), 5.94 (dd, J.sub.1=2.4 Hz, J.sub.2=18.8 Hz, 1H),
5.70 (d, J=8.0 Hz, 1H), 5.69 (d, J=0.8 Hz, 1H), 5.68 (s, 1H), 5.654
(d, J=1.2 Hz, 1H), 5.650 (s, 1H), 5.21 (dd, J=2.0, 5.2 Hz, 0.5H),
5.07 (dd, 2.0, 5.2 Hz, 0.5H), 4.42 (dd, J=5.6, 20.8 Hz, 1H), 4.14
(m, 2H), 1.223 (s, 9H), 1.220 (m, 9H); .sup.31P NMR (CD.sub.3OD)
4.92 (s); MS: m/z 698 [M+2-methylheptylamine]+.
Example 30
Preparation of Compound (30a)
##STR00196##
[0385] Preparation of (1-2): To a solution of 1-1 (313 mg; 0.55
mmol) in THF (8 mL) under Ar was added a solution of
triethylammonium bis(POM)phosphate in THF (prepared from
bis(POM)phosphate (215 mg; 1.2 equiv), THF (2 mL) and Et.sub.3N
(0.1 mL; 1.3 equiv)). The resulting mixture cooled in an ice-bath.
Diisopropylethyl amine (0.38 mL; 4 equiv) was added. BOP-Cl (280
mg; 2 equiv) and 3-nitro-1,2,4-triazole (125 mg; 2 equiv) was then
added. The reaction mixture was stirred at 0.degree. C. for 90
mins. The mixture was diluted with CH.sub.2Cl.sub.2 (60 mL) and
washed with saturated aq. NaHCO.sub.3 (2.times.10 mL) and brine.
The combined aqueous layers were back extracted with
CH.sub.ZCl.sub.2 (.about.20 mL). The combined organic extract was
dried (Na.sub.2SO.sub.4) and evaporated. The residue purified on
silica (25 g column) with CH.sub.ZCl.sub.2/i-PrOH solvent system
(2-10% gradient). Yield: 140 mg (27%).
[0386] Preparation of (30a): A solution of 1-2 (110 mg; 0.13 mmol)
in 80% aq. formic acid was heated at 35-37.degree. C. for 3 hours.
The mixture was evaporated to give an oily residue. The residue was
co-evaporated 2 times with toluene. Purification on a silica gel
column (10 g) with CH.sub.2Cl.sub.2/MeOH solvent system (4-10%
gradient) to afford compound 30a (46 mg, 59% yield). .sup.31P-NMR
(DMSO-.sub.6): .delta. -4.45. MS: m/z 646 (M+46-1).
Example 31
Preparation of Compound (31a)
##STR00197##
[0388] Preparation of (1-2): To a solution of 1-1 (313 mg; 0.55
mmol) in THF (8 mL) under Ar was added a solution of
triethylammonium bis(POM)phosphate in THF (prepared from
bis(POM)phosphate (215 mg; 1.2 equiv), THF (2 mL) and Et.sub.3N
(0.1 mL; 1.3 equiv)). The resulting mixture cooled in an ice-bath.
Diisopropylethyl amine (0.38 mL; 4 equiv) was added. BOP-Cl (280
mg; 2 equiv) and 3-nitro-1,2,4-triazole (125 mg; 2 equiv) was then
added. The reaction mixture was stirred at 0.degree. C. for 90
mins. The mixture was diluted with CH.sub.2Cl.sub.2 (60 mL) and
washed with saturated aq. NaHCO.sub.3 (2.times.10 mL) and brine.
The combined aqeuous layers were back extracted with
CH.sub.2Cl.sub.2 (.about.20 mL). The combined organic extract was
dried (Na.sub.2SO.sub.4), evaporated, and the residue purified on
silica (25 g column) with CH.sub.2Cl.sub.2/i-PrOH solvent system
(2-10% gradient). Yield: 154 mg (27%).
[0389] Preparation of (31a): A solution of 2-2 (68 mg; 0.08 mmol)
in 80% aq. formic acid was stirred at R.T. for 3 hours. The mixture
was evaporated to an oily residue. The residue was co-evaporated 2
times with toluene. Purification on a silica gel column (10 g) with
CH.sub.2Cl.sub.2/MeOH solvent system (4-10% gradient; target
compound eluted with 8% MeOH) afforded 31a (35 mg, 78% yield).
.sup.31P-NMR (DMSO-d.sub.6): .delta. -4.19. MS: m/z 580 (M-1), 646
(M+46-1), 550 (M-30-1).
Example 32
Preparation of Compound (32a)
##STR00198##
[0391] To a solution of 3-1 (71 mg; 0.26 mmol) in THF (4 mL) under
Ar was added triethylammonium bis(POM)phosphate (144 mg; 1.2
equiv), and the resulting mixture was cooled in an ice-bath, and
diisopropylethyl amine (0.18 mL; 4 equiv) was added. BOP-Cl (132
mg; 2 equiv) and 3-nitro-1,2,4-triazole (59 mg; 2 equiv) was then
added. The reaction mixture was stirred at 0.degree. C. for 1 hour.
The mixture was diluted with CH.sub.2Cl.sub.2 (50 mL) and washed
with saturated aq. NaHCO.sub.3 (2.times.10 mL) and brine. The
combined aqueous layers were back extracted with CH.sub.2Cl.sub.2
(.about.20 mL). The combined organic extract was dried
(Na.sub.2SO.sub.4), evaporated, and the residue was purified on
silica (10 g column) with CH.sub.2Cl.sub.2/MeOH solvent system
(4-10% gradient). Compound 32a was repurified by RP-HPLC (35-90% B;
A: water, B: MeOH). Yield 75 mg (50%). .sup.31P-NMR (DMSO-d.sub.6):
.delta. -4.14. MS: m/z 627 (M+46-1), 551 (M-30-1).
Example 33
Preparation of Compound (33a)
##STR00199##
[0393] Preparation of (4-2): To a solution of 4-1 (0.29 g; 0.5
mmol) in MeCN (8 mL) was added 5-ethylthio-1H-tetrazole in MeCN
(0.25 M; 2.4 mL; 1.2 equiv). BisSATE-phosphoramidate (0.24 g; 1.05
equiv.) in MeCN (1.5 mL) was added over 90 mins. The reaction
mixture was stirred for 4 hours at R.T., and then cooled to
-40.degree. C. MCPBA (0.23 g; 2 equiv.) in CH.sub.2Cl.sub.2 (3 mL)
was added. The mixture was allowed to warm to R.T. and diluted with
EtOAc (50 mL). The mixture was washed with 10% aq. NaHSO.sub.3
(2.times.10 mL), saturated aq. NaHCO.sub.3 (2.times.10 mL) and
brine. The mixture was then dried (Na.sub.2SO.sub.4). The
evaporated residue was purified on silica (10 g column) with
CH.sub.2Cl.sub.2/MeOH solvent system (4-10% gradient) to afford 4-2
(0.26 g, 55% yield).
[0394] Preparation of (33a): A solution of 4-2 (0.21 g; 0.22 mmol)
in 80% aq. AcOH (15 mL) was stirred 4 hours at R.T. The mixture was
evaporated and purified on silica (10 g column) with
CH.sub.2Cl.sub.2/MeOH solvent system (4-10% gradient). Yield: 0.13
g (90%). .sup.31P-NMR (DMSO-d.sub.6): .delta. -2.00. MS: m/z 686
(M+46-1).
Example 34
Preparation of Compounds (34a)-(34e)
##STR00200##
[0396] 1,2,4-Triazol (42 mg, 0.6 mmol) was suspended of dry
CH.sub.3CN (1 mL). Triethylamine was added (0.088 mL, 0.63 mmol),
and the mixture was vortexed to obtain a clear solution. After
addition of POCl.sub.3 (0.01 mL, 0.1 mmol), the mixture was
vortexed and left for 20 min. The mixture was then centrifugated.
The supernatant was added to the protected nucleoside (0.05 mmol),
and the mixture was kept at ambient temperature for 1 hour.
Tris(tetrabutylammonium) hydrogen pyrophosphate (180 mg, 0.2 mmol)
was added, and the mixture was kept for 2 hours at R.T. The
reaction was quenched with water, evaporated, dissolved in 80%
formic acid and left for 2 hours at R.T. Formic acid was
evaporated, and the residue dissolved in water (5 mL) and extracted
with EA (2.times.2 mL). The aqueous fraction was loaded onto column
HiLoad 16/10 with Q Sepharose High Performance (linear gradient of
NaCl from 0 to 1N in 50 mM TRIS-buffer (pH=7.5)). Fractions
containing the triphosphate were combined, concentrated and
desalted by RP HPLC on Synergy 4 micron Hydro-RP column
(Phenominex) using a linear gradient of methanol from 0 to 20% in
50 mM triethylammonium acetate buffer (pH 7.5) for elution. The
following compounds shown in Table 1 were synthesized according
this procedure:
TABLE-US-00001 TABLE 1 Triphosphates obtained from Example 34
.sup.31P NMR .sup.31P NMR .sup.31P NMR MS Compound P.alpha. P.beta.
P.gamma. (M.sup.-) ##STR00201## -11.31 d -20.82 t -5.48 d 550.2
##STR00202## -9.13 d -8.18 t -2.85 d 548.2 ##STR00203## -10.95 d
-20.62 bs -5.37 bs 552.2 ##STR00204## -11.24 d -20.82 t -5.48 d
554.2 ##STR00205## -12.06 d -20.97 t -5.69 d 549.2
Example 35
Preparation of Compound (35a)
##STR00206##
[0398] 1,2,4-Triazol (42 mg, 0.6 mmol) was suspended in dry
CH.sub.3CN (1 mL). Triethylamine was added (0.088 mL, 0.63 mmol),
and the mixture was vortexed to obtain a clear solution. After
addition of POCl.sub.3 (0.01 mL, 0.1 mmol), the mixture was
vortexed and left for 20 mins. The mixture was centrifugated, and
the supernatant was added to the protected nucleoside (0.05 mmol).
The mixture was kept at ambient temperature for 1 hour.
Tris(tetrabutylammonium) hydrogen pyrophosphate (180 mg, 0.2 mmol)
was added, and the mixture was kept for 2 hours at R.T. The
reaction was quenched with water, evaporated, dissolved in ammonium
hydroxide and left for 2 hours at R.T. The solvent was evaporated,
and the residue dissolved in water (10 mL). The mixture was loaded
onto a column HiLoad 16/10 with Q Sepharose High Performance.
Separation was done in linear gradient of NaCl from 0 to 1N in 50
mM TRIS-buffer (pH7.5). The fractions containing the product were
combined, concentrated and desalted by RP HPLC on Synergy 4 micron
Hydro-RP column (Phenominex). A linear gradient of methanol from 0
to 20% in 50 mM triethylammonium acetate buffer (pH 7.5) was used
for elution. MS (M-1): 532.1. .sup.31P-NMR (.delta. ppm): -5.12
(d), -11.31 (d) and -20.43 (t).
Example 36
Preparation of Compounds (36a)-(36d)
##STR00207##
[0400] 2'-Deoxy-2'-fluoro-4'-alkyl-cytidine (0.09 mmol) was
dissolved in the mixture of DMF (5 mL) and N,N'-dimethylacetate in
DMF (0.110 mL, 0.9 mmol). The reaction mixture left at R.T.
overnight. The solvent was evaporated, and the residue purified by
flash chromatography in gradient of methanol in DCM from 3% to 20%.
The N-Protected nucleoside was concentrated in vacuum, dried and
dissolved in dry trimethylphosphate (0.7 mL). The solution was
cooled to 4.degree. C. and POCl.sub.3 (0.017 mL, 0.18 mmol) was
added. In 1 hour, tributylamine (0.102 mL, 0.3 mmol) was added at
R.T. Tributylammonium pyrophosphate (156 mg, 0.34 mmol) was then
added. Dry DMF (about 0.100 mL) was added to solubilize
pyrophosphate. After 2 hours, the reaction was quenched with
TEAB-buffer. The product was isolated by ion-exchange
chromatography on AKTA Explorer as described in Example 35. The
fractions containing the product were concentrated and treated with
NH.sub.4OH for 2 hours at R.T. The product was desalted by RP HPLC
as described in Example 35.
TABLE-US-00002 TABLE 2 Triphosphates obtained from Example 36
.sup.31P NMR .sup.31P NMR .sup.31P NMR MS P.alpha. P.beta. P.gamma.
(M.sup.-) ##STR00208## -11.38 bs -22.88 bs -7.62 bs 512.1
##STR00209## -11.49 bs -20.41 bs -5.34 bs 510.0 ##STR00210## -11.96
bs -22.07 t -5.66 d 508.3 ##STR00211## -11.90 d -23.23 t -10.66 d
514.0 ##STR00212## -11.77 d -23.05 t -9.70 s 529.9 ##STR00213##
-11.74 d -23.37 t -10.85 d 539.2 ##STR00214## -11.87 d -23.32 t
-10.83 d 523.9 ##STR00215## -11.48 d -23.26 t -10.63 d 526.1
##STR00216## -11.67 d -23.22 t -10.77 d 554.1 ##STR00217## -11.97 d
-23.34 t -10.92 d 523.9
Example 37
Preparation of Compounds (37a)
##STR00218##
[0402] Compound 37a was synthesized by reaction of
phosphor(tris-triazolide) with 4'-ethyl-2'-deoxy-2'-fluoro-uridine
as described Examples 34 and 35. MS (M-1): 513.1. .sup.31P-NMR
(.delta. ppm): -9.43 (bs), -11.68 (d) and -23.09 (bs).
Example 38
Preparation of Compounds (38a)
##STR00219##
[0404] The starting nucleoside (15 mg, 0.05 mmol) was dissolved in
dry trimethylphosphate (3 mL). The solution was cooled to 4.degree.
C. POCl.sub.3 (0.013 mL, 0.125 mmol) was added, followed by
pyridine (0.01 mL, 0.125 mmol). In 1 hour, tributylamine (0.035 mL,
0.125 mmol) was added at R.T. followed by tributylammonium
pyrophosphate (156 mg, 0.34 mmol). Dry DMF (about 0.100 mL) was
added to solubilize pyrophosphate. In 2 hours, the reaction was
quenched with TEAB-buffer. The product was isolated by ion-exchange
chromatography on AKTA Explorer as described in Example 35. The
fractions containing the product were concentrated and treated with
NH.sub.4OH for 2 hours at R.T. The product was desalted by RP HPLC
as described in Example 35. MS (M-1): 529.9. .sup.31P-NMR (.delta.
ppm): -9.42(d), -11.59(d) and -23.03(t).
Example 39
Preparation of Compound (40a)
##STR00220## ##STR00221##
[0406] Preparation of (40-2): To a solution of 40-1 (50.0 g, 205
mmol) in pyridine (250 mL) was added DMTrCl (75.0 g, 225.0 mmol).
The solution was stirred at R.T. for 15 hours. MeOH (120 mL) was
added, and the mixture was concentrated to dryness under reduced
pressure. The residue was dissolved in EA and washed with water.
The organic layer was dried over Na.sub.2SO.sub.4 and concentrated
to give the crude 5'-O-DMTr intermediate (80.52 g) as a light
yellow solid. The intermediate was dissolved in anhydrous DMF (300
mL), and K.sub.2CO.sub.3 (80.52 g, 583.2 mmol) was added followed
by PMBCl (31.7 g, 109.2 mmol). The mixture was stirred at R.T.
overnight. The reaction was diluted with EA and washed with brine.
The organic phase was dried over Na.sub.2SO.sub.4 and concentrated
to give crude 5'-O-DMTr-N3-PMB FdU (98.8 g) as a light yellow
solid. The solid was dissolved in DMF (300 mL), and NaH (10.42 g,
260.5 mmol) was added followed by BnBr (73.8 g, 434.2 mmol). The
reaction was stirred at R.T. overnight and then was quenched with
water. The solution was diluted with EA and washed with brine. The
organic phase was dried over Na.sub.2SO.sub.4 and concentrated to
give the crude fully blocked FdU intermediate, which was purified
on a silica gel column (PE:EA=10:1 to 3:1) to the pure fully
blocked FdU (101.1 g). The intermediate was treated with 80% HOAc
(900 mL) at R.T. overnight, and the solvent was removed. The
residue was purified on a silica gel column to give 40-2 as a white
foam (42.1 g, 30.2% for 4 steps).
[0407] Preparation of (40-3): To a solution of 40-2 (42.1 g, 92.6
mmol) in anhydrous CH.sub.3CN (300 mL) was added IBX (28.5 g, 121.7
mmol) at R.T. The reaction mixture was refluxed for 1 hour and then
cooled to 0.degree. C. The precipitate was filtered-off, and the
filtrate was concentrated to give the crude aldehyde (39.22 g) as a
yellow solid. To a solution of the aldehyde (39.22 g) in
1,4-dioxane (250 mL) was added 37% CH.sub.2O (28.1 mL, 345.6 mmol)
and 2N NaOH aqueous solution (86.4 mL, 172.8 mmol). The mixture was
stirred at R.T. for 2 hours and then neutralized with AcOH to pH=7.
EtOH (200 mL) and NaBH.sub.4 (19.7 g, 518.6 mmol) were added,
stirred at R.T. for 30 mins. The mixture was quenched with
saturated aqueous NH.sub.4Cl, and extracted with EA. The organic
layer was dried over Na.sub.2SO.sub.4 and concentrated. The residue
was purified by silica gel column chromatography (PE:EA=4:1 to 2:1)
to give 40-3 (25.5 g, 55.7%) as a white solid.
[0408] Preparation of (4-4): To a stirred solution of 40-3 (25.5 g,
52.5 mmol) in anhydrous pyridine (150 mL) and anhydrous CH.sub.3CN
(150 mL) was added BzCl (6.6 g, 52.47 mmol) dropwise at 0.degree.
C. The mixture was stirred at R.T. for 14 hours. The reaction was
quenched with H.sub.2O, and the solution was concentrated. The
residue was dissolved in EA and washed with saturated NaHCO.sub.3.
The organic layer was dried over Na.sub.2SO.sub.4 and concentrated.
The residue was purified on a silica gel column (PE/EA=5:4) to give
the mono-Bz protected intermediate (18.1 g, 60.0%) as a white foam.
To a stirred solution of this intermediate (18.1 g, 30.68 mmol) in
DMF (100 mL) were added Cs.sub.2CO.sub.3 (30.0 g, 92.03 mmol) and
BnBr (10.4 g, 61.36 mmol). The mixture was stirred at R.T.
overnight. The reaction was quenched with saturated NH.sub.4Cl aq.,
extracted with EA and washed with brine. The solvent was removed to
give crude 40-4 (19.3 g, 95.1%) as a light yellow solid.
[0409] Preparation of (40-5): To a stirred solution of 40-4 (19.3
g, 28.4 mmol) in anhydrous MeOH (230 mL) was added NaOMe (24.9 g,
460 mmol) at R.T. The mixture was stirred for 1 hour. The reaction
was quenched with AcOH (10 mL) and concentrated. The residue was
purified on a silica gel column (PE/EA=1/2) to afford 40-5 (11.2 g,
54.0%) as a white solid.
[0410] Preparation of (40-6): To a stirred solution of compound
40-5 (200 mg, 0.347 mmol) in anhydrous DCM (5 mL) was added DMP
(168 mg, 0.674 mmol) at R.T. The mixture was stirred at R.T. for 2
hours. The solvent was removed, and the residue was purified on a
silica gel column (PE:EA=5:1 to 1:1) to give the aldehyde crude as
a light yellow solid (200 mg). To a stirred solution of the
aldehyde (200 mg) in anhydrous THF (5 mL) was added MeMgBr (1.0 mL,
1.01 mmol) at -78.degree. C. The mixture was stirred at -78.degree.
C. for 1 hour. The reaction was quenched with saturated NH.sub.4Cl
aq. and extracted with EA. The concentrated organic phase was
purified by column chromatography (PE:EA=5:1 to 1:1) to give 40-6
(a mixture of stereomers, 135 mg, 65%) as a white solid.
[0411] Preparation of (40-7): To a stirred solution of DAST (1.64
g, 10.17 mmol) in anhydrous toluene (40 mL) was added dropwise a
solution of compound 40-6 (1.2 g, 2.03 mmol) at -78.degree. C. The
mixture was stirred at -78.degree. C. for 30 mins. The solution was
warmed to 60.degree. C. slowly and stirring was continued
overnight. The mixture was poured into a saturated Na.sub.2CO.sub.3
solution. The concentrated organic phase was concentrated and
purified on a silica gel column (PE:EA=10:1 to 3:1) to afford 40-7
as a white solid (1.08 g, 83.88%). .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 7.87 (d, J=8.4 Hz, 1H), 7.27-7.37 (m, 12H), 6.82-6.84
(m, 2H), 6.14 (d, J=16.8, 2.0 Hz, 1H), 5.18-5.50 (m, 4H), 4.96 (s,
2H), 4.45-4.88 (m, 7H), 3.67-3.89 (m, 5H).
[0412] Preparation of (4-8): A mixture of compound 40-7 (0.91 g,
1.54 mmol) and CAN (2.53 g, 4.61 mmol) in a 3:1 solution of
MeCN:water (10 m L) was stirred at R.T. overnight. Brine (10 mL)
was added, and the mixture was extracted with EA. The combined
organic extracts were dried and evaporated under reduced pressure.
Purification by chromatography on silica gel column with PE:EA=10:1
to 2:1 afforded 40-8 as a yellow solid (305 mg, 41.96%).
[0413] Preparation of (40-9): To a stirred solution of 40-8 (350
mg, 0.74 mmol) in anhydrous MeCN (8 mL) were added TPSCl (449 mg,
1.48 mmol), DMAP (180 mg, 1.48 mmol) and TEA (374 mg, 3.70 mmol) at
R.T. The mixture was stirred at R.T. overnight. NH.sub.4H (15 mL)
was added, and the mixture was stirred for 2 hours. The solvent was
removed, and the residue was purified on a silica gel column with
PE:EA=8:1 to 1:1 to afford the crude (380 mg crude), which was
dissolved in anhydrous DCM (10 mL). A mixture of MMTrCl (695 mg,
2.25 mmol) and AgNO.sub.3 (380 mg, 2.25 mmol) was added at R.T.,
and the mixture was stirred at R.T. overnight. The solid was
filtered off and washed with DCM. The filtrate was washed with
brine and dried over Na.sub.2SO.sub.4. The concentrated organic
phase was purified on a silica gel column (PE:EA=8:1 to 2:1) to
afford 40-9 as a yellow solid (460 mg, 81.33%).
[0414] Preparation of (40-1): To a stirred solution of compound
40-9 (450 mg, 0.61 mmol) in acetone were added ammonium formate
(1.29 g, 20.6 mmol, in portions) and 10% palladium on carbon (1.0
g). The mixture was refluxed for 12 h. The catalyst was filtered
off and washed with acetone. The filtrate was diluted with EA and
washed with brine. The concentrated organic phase was purified by
column chromatography (DCM:MeOH=100:1 to 15:1) to afford 40-10 as a
white solid (250 mg, 72.8%). .sup.1H NMR (DMSO-d6, 400 M Hz)
.delta. 8.56 (s, 1H), 7.73 (d, J=7.6 Hz, 1H), 7.14-7.28 (m, 12H),
6.84 (d, J=8.8 Hz, 2H), 6.30 (d, J=7.6 Hz, 1H), 6.03-6.08 (m, 1H),
5.84 (d, J=5.2 Hz, 1H), 5.33-5.35 (m, 1H), 4.97-5.18 (m, 1H),
4.86-4.90 (m, 1H), 4.34 (d, J=4.4 Hz, 1H), 3.72 (s, 3H), 3.54-3.57
(m, 2H), 1.28 (dd, J.sub.1=6.4 Hz, J.sub.2=25.6 Hz, 3H). ESI-MS:
m/z 563.50 [M+H].sup.+.
[0415] Preparation of (40a): 40-10 (101 mg, 0.179 mmol) was
dissolved in 80% HOAc (20 mL) at R.T. The mixture was stirred at
50.degree. C. for 5 hours. The solvent was removed, and the residue
was co-evaporated with toluene twice. The residue was purified by
column chromatography (DCM:MeOH=100:1 to 10:1) to afford 40a as a
white solid (36.6 mg, 70.26%). .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 7.98 (d, J=7.6 Hz, 1H), 6.20-6.24 (m, 1H), 5.92 (d, J=7.2
Hz, 1H), 5.17-5.30 (m, 1H), 4.99-5.14 (m, 1H), 4.51-4.86 (m, 1H),
3.78 (d, J=1.6 Hz, 2H), 1.35-1.43 (m, 3H). ESI-MS: m/z 291.84
[M+H].sup.+, 582.81 [2M+H].sup.+.
Example 40
Preparation of Compound (41a)
##STR00222##
[0417] Preparation of (41-2): To a solution of 41-1 (3 g, 4.8 mmol)
in anhydrous DCM (50 mL) were added BzCl (1.3 g, 9.6 mmol), DMAP
(1.1 g, 9.6 mmol) and NEt.sub.3 (4 mL) at R.T. The reaction was
stirred at R.T. for 2 hours. Water was added, and the reaction was
stirred for another 1 hour. The mixture was diluted with DCM (150
mL) and washed with water, 0.1 M HCl and saturated aqueous
NaHCO.sub.3. The solvent was removed, and the crude product was
purified by silica gel column chromatography (25% EtOAc in PE) to
give 41-2 as a yellow solid (2.8 g, 80.0%).
[0418] Preparation of (41-3): A mixture of 41-2 (2.6 g, 3.6 mmol)
and Pd(OAc).sub.2 (100 mg) in DCM (50 mL) was suspended in a
solution of CH.sub.2N2 in Et.sub.2O (generated by standard
procedure, 350 mL) at -78.degree. C. The reaction was stirred to
R.T. overnight. The mixture was quenched with HOAc, and the
reaction was stirred for another 1 hour. The mixture was diluted
with EtOAc (150 mL) and washed with water and saturated aqueous
NaHCO.sub.3. The solvent was removed, and the crude was dissolved
in NH.sub.3.MeOH (sat., 100 mL). The reaction was stirred to R.T.
overnight. The crude product was purified by silica gel column
chromatography (25% EtOAc in PE) to give 41-3 as a yellow solid
(800 mg, 35.2%).
[0419] Preparation of (41-4): To a solution of 41-3 (800 mg, 1.3
mmol) in anhydrous CH.sub.3CN (50 mL) were added TPSCl (755 mg, 2.5
mmol), DMAP (305 mg, 2.5 mmol) and NEt.sub.3 (400 mg, 4 mmol) at
R.T. The reaction was stirred at R.T. for 2 hours. NH.sub.4OH (25
mL) was added, and the reaction was stirred for another 1 hour. The
mixture was diluted with DCM (150 mL) and washed with water, 0.1 M
HCl and saturated aqueous NaHCO.sub.3. The solvent was removed, and
the crude product was purified by silica gel column chromatography
(25% EtOAc in PE) to give 41-4 as a yellow solid (340 mg,
42.5%).
[0420] Preparation of (41a): To a solution of 41-4 (200.0 mg) in
MeOH (10 mL) was added NH.sub.4F (600 mg). The reaction was
refluxed for 24 hours. The solvent was removed, and the residue was
purified by column chromatography on silica gel (DCM:MeOH=15:1) to
give 41a (50.0 mg, 55.9%) as a white solid. .sup.1H NMR
(CD.sub.3OD, 400 M Hz) .delta. 8.13 (d, J=7.6 Hz, 1H), 6.01 (dd,
J.sub.1=2.4 Hz, J.sub.2=15.6 Hz, 1H), 5.85 (d, J=7.6 Hz, 1H),
5.04-4.89 (m, 1H), 4.52 (dd, J.sub.1=5.2 Hz, J.sub.2=19.6 Hz, 1H),
3.66 (s, 2H), 1.00-0.94 (m, 1H), 0.54-0.30 (m, 4H); ESI-MS: m/z
285.82 [M+H].sup.+, 570.84 [2M+H].sup.+.
Example 41
Preparation of Compound (42a)
##STR00223## ##STR00224##
[0422] Preparation of (42-2): To a solution of 42-1 (50 g, 203
mmol) in anhydrous pyridine (200 mL) was added TBDPSCl (83.7 g, 304
mmol, 1.5 eq). The reaction was stirred overnight at R.T. The
solution was concentrated under reduced pressure to give a syrup,
which was partitioned between ethyl acetate and water. The organic
layer was separated, washed with brine, dried over magnesium
sulfate and concentrated to give the 5'-OTBDPS ether as a white
foam (94 g). The crude ether was dissolved in anhydrous DCM (300
mL), and silver nitrate (66.03 g, 388.4 mmol, 2.0 eq) and collidine
(235 mL, 1.94 mol, 10 eq) were added. The mixture was stirred at
R.T., and MMTrCl (239.3 g, 776.8 mmol, 4 eq) was added. After being
stirred overnight at R.T., the mixture was filtered through Celite
and filtrate was diluted with MTBE. The solution was washed
successively with 1M citric acid, diluted brine and 5% sodium
bicarbonate. The organic solution was dried over sodium sulfate and
concentrated under vacuum to give the fully protected intermediate
as a yellow foam. The crude intermediate was dissolved in anhydrous
THF (250 mL) and treated with TBAF (60 g, 233 mmol, 1.2 eq). The
mixture was stirred for 2 hours at R.T., and the solvent was
removed under reduced pressure. The residue was taken into ethyl
acetate and washed brine. After drying over magnesium sulfate, the
solvent was removed in vacuo. The residue was purified by column
chromatography (PE:EA=5:1 to 1:1) to give 42-2 as a white foam (91
g, 86.4%).
[0423] Preparation of (42-3): To a solution of 42-2 (13.5 g, 26
mmol) in DCM (100 mL) was added pyridine (6.17 mL, 78 mmol, 3 eq).
The solution was cooled to 0.degree. C. and Dess-Martin periodinane
(33.8 g, 78 mmol, 3 eq) was added. The mixture was stirred for 4
hours at R.T. and quenched by the addition of a 4%
Na.sub.2S.sub.2O.sub.3/4% sodium bicarbonate aqueous solution (to
pH 6, -150 mL). The mixture was stirred for another 15 mins. The
organic layer was separated, washed with diluted brine and
concentrated under reduced pressure. The residue was dissolved in
dioxane (100 mL), and the solution was treated with 37% aqueous
formaldehyde (21.2 g, 10 eq) and 2N aqueous sodium hydroxide (10
eq). The reaction mixture was stirred at R.T. overnight. The
reaction was quenched with saturated NH.sub.4CI (.about.150 mL),
and the mixture was concentrated under reduced pressure. The
residue was partitioned between ethyl acetate and 5% sodium
bicarbonate. The organic phase was separated, washed with brine,
dried over magnesium sulfate and concentrated. The residue was
purified by column chromatography (MeOH:DCM=100:1-50:1) to give
42-3 as a white foam (9.2 g, 83.6%).
[0424] Preparation of (42-4): 42-3 (23 g, 42.0 mmol) was
co-evaporated with toluene twice. The residue was dissolved in
anhydrous DCM (250 mL) and pyridine (20 mL). The solution was
cooled to -35.degree. C. Triflic anhydride (24.9 g, 88.1 mmol, 2.1
eq) was added dropwise over 10 mins. At this temperature, the
reaction was stirred for 40 mins and then was quenched with water
(50 mL) at 0.degree. C. The mixture was stirred 30 mins, and
extracted with EA (150 mL.times.2). The organic phase was dried
over Na.sub.2SO.sub.4, and filtered through a silica gel pad. The
filtrate was concentrated under reduced pressure. The residue was
purified by column chromatography (PE:EA=100:1-1:1) to give 42-4 as
a brown foam (30.0 g, 88.3%).
[0425] Preparation of (42-5): 42-4 (30 g, 36.9 mmol) was
co-evaporated twice with toluene and dissolved in anhydrous DMF
(150 mL). The solution was cooled to 0.degree. C., and treated with
sodium hydride (60% in mineral oil; 1.5 g, 40.6 mmol). The reaction
was stirred at R.T. for 1 h. Lithium chloride (4.6 g, 110.7 mmol, 3
eq) was added. Stirring was continued for 2 hours when LCMS
indicated complete conversion of the anhydro triflate intermediate
to anhydro-chloro compound. The mixture was taken into 100 mL of
half saturated ammonium chloride and ethyl acetate. The organic
phase was separated, washed with diluted brine and concentrated
under reduced pressure. The residue was dissolved in THF (150 mL),
and the solution was treated with 1N aqueous sodium hydroxide
(.about.41 mL, 40.1 mmol, 1.1 eq). The mixture was stirred at R.T.
for 1 h. The reaction was diluted with half saturated sodium
bicarbonate (.about.60 mL) and extracted with EA. The organic phase
was dried (magnesium sulfate) and concentrated under reduced
pressure. The residue was purified by column chromatography
(DCM:MeOH=300:1-60:1) to give 42-5 as a yellow foam (18.3 g,
87.6%).
[0426] Preparation of (42-6): To a solution of 42-5 (18.3 g, 32.33
mmol) in anhydrous DCM (150 mL) was added TBSCl (17.7 g, 64.6 mmol)
and imidazole (6.6 g, 97 mmol). The reaction was stirred overnight
at R.T. The reaction was diluted with water and extracted with DCM.
The organic layer was separated, washed with brine, dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by
column chromatography (DCM:MeOH=300:1-80:1) to give 42-6 as a white
foam (18.4 g, 83.7%).
[0427] Preparation of (42-7): A solution of 42-6 (18.4 g, 27.1
mmol), DMAP (6.6 g, 54.0 mmol) and TEA (5.4 g, 54.0 mmol) in MeCN
(450 mL) was treated with 2,4,6-triispropylbenzenesulfonyl chloride
(16.3 g, 54.0 mmol). The mixture was stirred at R.T. for 3 hours.
NH.sub.4OH (70 mL) was added, and the mixture was stirred for 2
hours. The solution was evaporated under reduced pressure, and the
residue was purified on a silica gel column (DCM/MeOH=100:1 to
15:1) to give the crude (18.0 g). The crude was dissolved in
anhydrous DCM (150 mL). Collidine (8.1 g, 66.3 mmol, 2.5 eq),
silver nitrate (4.5 g, 26.5 mmol, 1.0 eq) and DMTrCl (13.4 g, 39.7
mmol, 1.5 eq) were added. The reaction was stirred overnight at
R.T. The mixture was filtered through Celite. The filtrate was
washed with brine and extracted with DCM. The organic layer was
separated, dried over Na.sub.2SO.sub.4 and concentrated. The
residue was purified by column chromatography (PE:EA=60:1-3:1) as a
yellow foam. The foam was dissolved in THF (150 mL) and TBAF (10.4
g, 39.7 mmol, 1.5 eq) was added. The reaction was stirred at R.T.
After being concentrated, the mixture was washed with brine and
extracted with EA. The organic layer was separated, dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by
column chromatography (PE:EA=60:1-EA) to give 42-7 as a yellow foam
(21.3 g, 92.4%).
[0428] Preparation of (42-8): To a solution of 42-7 (2.0 g, 2.3
mmol) in anhydrous DCM (20 mL) was added Dess-Martin periodinane
(1.95 g, 4.6 mmol) at 0.degree. C. under nitrogen. The reaction was
stirred at R.T. for 5 hours. The mixture was diluted with EtOAc
(100 mL), and washed with a mixture of saturated aqueous
Na.sub.2S.sub.2O.sub.3 and saturated aqueous NaHCO.sub.3. The crude
product was purified by column chromatography on silica gel
(PE:EtOAc=2:1) to give 42-8 (1.8 g, 90%) as a yellow solid.
[0429] Preparation of (42-9): To a solution of tetramethyl
methylenediphosphonate (390 mg, 1.68 mmol) in anhydrous THF (10 mL)
was added NaH (84 mg, 2.1 mmol) at 0.degree. C. under nitrogen. The
reaction was stirred at 0.degree. C. for 30 min. A solution of 42-8
(1.2 g, 1.4 mmol) in anhydrous THF (10 mL) was added dropwise at
0.degree. C. The mixture was stirred at R.T. for 1 h. The reaction
was quenched with saturated aqueous NH.sub.4Cl, and the crude
product was purified by column chromatography on silica gel
(DCM:MeOH=150:1) to give 42-9 (1.2 g, 88.2%) as a yellow solid.
.sup.1H NMR (DMSO-d6, 400 M Hz) .delta. 8.51 (s, 1H), 7.46-7.09 (m,
22H), 6.88-6.82 (m, 6H), 6.62 (q, J.sub.1=17.2 Hz, J.sub.2=22.4 Hz,
1H), 6.12 (d, J=7.2 Hz, 1H), 5.86-5.75 (m, 2H), 5.43 (d, J=25.2 Hz,
1H), 4.63 (dd, J=4.8 Hz, J.sub.2=21.2 Hz, 1H), 4.45 (d, J=12.0 Hz,
1H), 3.94 (d, J=12.0 Hz, 1H), 3.72 (s, 9H), 3.53 (q, J.sub.1=11.2
Hz, J.sub.2=16.0 Hz, 6H); ESI-MS: m/z 971.59 [M+H].sup.+.
[0430] Preparation of (42a): A solution of 42-9 (300 mg) in 80%
HOAc (26 mL) was stirred at 80-90.degree. C. for 2 h. The solvent
was removed, and the crude product was purified by column
chromatography on silica gel (DCM:MeOH 20:1) to give 42a (70 mg,
57%) as a white solid. .sup.1H NMR (DMSO-d6, 400 M Hz) .delta. 7.61
(d, J=7.6 Hz, 1H), 7.35 (d, J=15.2 Hz, 2H), 6.72 (q, J.sub.1=17.6
Hz, J.sub.2=24.4 Hz, 1H), 6.23 (d, J=6.0 Hz, 1H), 5.99-5.85 (m,
2H), 5.74 (q, J=7.2 Hz, 1H), 5.37-5.21 (m, 1H), 4.69-4.61 (m, 1H),
3.96 (d, J=12.4 Hz, 1H), 3.82 (d, J=12.0 Hz, 1H), 6.72 (q,
J.sub.1=5.2 Hz, J.sub.2=10.8 Hz, 6H); ESI-MS: m/z 397.81
[M+H].sup.+.
Example 42
Preparation of Compound (43a)
##STR00225##
[0432] Preparation of (43-2): To a stirred solution of 43-1 (3.8 g,
6.6 mmol) in anhydrous DMF (100 mL) was added NaH (2.2 g) followed
by CH.sub.3I (9.3 g, 66 mmol) at 0.degree. C. Stirring was
continued at R.T. overnight. The reaction was quenched with
saturated NH.sub.4Cl aq. The mixture was diluted with EA and washed
with brine. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated. The residue was purified by silica gel column
chromatography (PE:EA=2:1) to give 43-2 (3.0 g, 70%) as a white
solid.
[0433] Preparation of (43-3): A mixture of 43-2 (3.0 g, 5.1 mmol)
and CAN (5.56 g, 10.2 mmol) in a 3:1 solution of MeCN:Water (16 mL)
was stirred at R.T. overnight. The solution was diluted with brine
(10 mL) and was extracted with EA. The combined organic extracts
were dried and evaporated under reduced pressure. Purification by
chromatography on silica (PE:EA=1:1) gave 43-3 as a yellow solid
(1.71 g, 72%).
[0434] Preparation of (43-4): To a stirred solution of 43-3 (1.7 g,
3.6 mmol) in anhydrous MeCN (50 mL) were added TPSCl (2.2 g, 7.2
mmol), DMAP (880 mg, 7.2 mmol) and TEA (1.1 g, 10.8 mmol) at R.T.
The mixture was stirred at R.T. overnight. NH.sub.4OH (25 mL) was
added, and the mixture was stirred for 2 hours. The solvent was
removed, and the residue was purified on a silica gel column
(PE:EA=8:1 to 2:1) to give the intermediate (1.4 g). The
intermediate was dissolved in anhydrous DCM (30 mL), and MMTrCl
(1.6 g, 5.2 mmol), AgNO.sub.3 (1.4 g, 7.8 mmol) and collidine (1.57
g, 13 mmol) were added. The mixture was stirred at R.T. overnight.
The solid was filtered off and washed with DCM. The filtrate was
washed with brine and dried over Na.sub.2SO.sub.4. The concentrated
organic phase was purified on a silica gel column (PE:EA=3:2) to
give 43-4 (1.1 g, 57.9%) as a white solid.
[0435] Preparation of (43-5): To a stirred solution of 43-4 (550
mg, 0.74 mmol) in acetone were added ammonium formate (1.0 g, 15.8
mmol, in portions) and 10% palladium on carbon (1.0 g). The mixture
was refluxed for 48 hours. The catalyst was filtered off and washed
with the acetone. The filtrate was diluted with EA, washed with
brine and dried. The concentrated organic phase was purified by
column chromatography (DCM:MeOH=50:1) to give 43-5 (330 mg,
72%).
[0436] Preparation of (43a): 43-5 (200 mg, 0.36 mmol) was dissolved
in 80% CH.sub.3COOH (20 mL) at R.T. The mixture was stirred at
60.degree. C. for 12 hours. The solvent was removed. The residue
was purified by column chromatography (DCM:MeOH=10:1), and the
resulting solid was washed with DCM to give pure 43a as a white
solid (44 mg, 42%). .sup.1H NMR (CD3OD, 400 MHz) .delta. 8.02 (d,
J=7.2 Hz, 1H), 6.14 (dd, J.sub.1=3.6 Hz, J.sub.2=15.2 Hz, 1H), 5.88
(d, J=7.2 Hz, 1H), 5.10 (ddd, J.sub.1=4.0 Hz, J.sub.2=5.2 Hz,
J.sub.3=53.6 Hz, 1H), 4.47 (dd, J.sub.1=5.2 Hz, J.sub.2=14.8 Hz,
1H), 3.84 (d, J=12.0 Hz, 1H), 3.70 (d, J=12.0 Hz, 1H), 3.58-3.64
(m, 2H), 3.36 (s, 3H). ESI-MS: m/z 290 [M+H].sup.+.
Example 43
Preparation of Compound (44a)
##STR00226##
[0438] To a solution of triethylammonium bis(POM)phosphate (0.3
mmol, prepared from 100 mg of bis(POM)phosphate and 50 .mu.L of
Et.sub.3N) in THF (3 mL) was added nucleoside 44-1 (150 mg; 0.26
mmol). The mixture was cooled in ice-bath. Diisopropylethyl amine
(0.18 mL; 4 equiv) was added then, followed by BOP-Cl (132 mg; 2
equiv) and 3-nitro-1,2,4-triazole (59 mg; 2 equiv). The reaction
mixture was stirred at 0.degree. C. for 90 mins., and then diluted
with CH.sub.2Cl.sub.2 (30 mL) and washed with saturated aq.
NaHCO.sub.3 and brine. The combined aqueous layers were back
extracted with CH.sub.2Cl.sub.2. The combined organic extract was
dried (Na.sub.2SO.sub.4), evaporated, and the residue purified on
silica (10 g column) with CH.sub.2Cl.sub.2/i-PrOH solvent system
(3-10% gradient). The obtained mixture of products were treated for
30 mins at 35.degree. C. with 80% aq. HCOOH, and then evaporated
and coevaporated with toluene. The evaporated residue was purified
on silica (10 g column) with CH.sub.2Cl.sub.2/MeOH solvent system
(5-10% gradient) to obtain 44a (8 mg, 5%). .sup.31P-NMR
(DMSO-d.sub.6): .delta. -5.07. MS: m/z=668 (M+46-1).
Example 44
Preparation of Compound (45a)
##STR00227##
[0440] Preparation of (45-2): To a solution of triethylammonium
bis(POM)phosphate (0.7 mmol, prepared from 233 mg of
bis(POM)phosphate and 0.1 mL of Et.sub.3N) in THF (8 mL) was added
nucleoside 45-1 (253 mg; 0.42 mmol), followed by diisopropylethyl
amine (0.36 mL; 5 equiv), BOP-Cl (268 mg; 2.5 equiv) and
3-nitro-1,2,4-triazole (120 mg; 2.5 equiv). The reaction mixture
was stirred at R.T. for 2 hours. The mixture was diluted with
CH.sub.2Cl.sub.2 (40 mL) and washed with saturated aq. NaHCO.sub.3
and brine. The combined aqueous layers were back extracted with
CH.sub.2Cl.sub.2. The combined organic extract was dried
(Na.sub.2SO.sub.4), evaporated, and the residue was purified on
silica (10 g column) with hexanes/EtOAc solvent system (40-100%
gradient) to yield 45a (180 mg, 47%).
[0441] Preparation of (45a): A solution of compound 45-2 (0.12 g;
0.13 mmol) in 80% aq. HCOOH (8 mL) was stirred 30 mins. at R.T. The
mixture was evaporated, coevaporated with toluene and purified on
silica (10 g column) with CH.sub.2Cl.sub.2/MeOH solvent system
(4-10% gradient) to yield 45a (55 mg, 70%). .sup.31P-NMR
(DMSO-d.sub.6): .delta. -4.36. MS: m/z=647 (M+46-1).
Example 45
Preparation of Compound (46a)
##STR00228##
[0443] Preparation of (46-2): A mixture of 46-1 (170 mg; 0.3 mmol)
in pyridine (3 mL) and isobutyric anhydride (0.1 mL; 2 equiv) was
stirred o/n at R.T. The mixture was concentrated, and the residue
was partitioned between EtOAc (30 mL) and saturated aq.
NaHCO.sub.3. The organic layer was washed with water, brine and
dried (Na.sub.2SO.sub.4). The residue was purified on silica (10 g
column) with a hexanes/EtOAc solvent system (30 to 100% gradient)
to afford 46-2 (180 mg, 85%).
[0444] Preparation of (46a): A solution of 46-2 (0.18 g; 0.25 mmol)
in 80% aq. HCOOH (5 mL) was heated for 3 hours at 36.degree. C. The
mixture was then evaporated, coevaporated with toluene and purified
on silica (10 g column) with a CH.sub.2Cl.sub.2/MeOH solvent system
(4-10% gradient) to afford 46a (75 mg, 70%). MS: m/z=434 (M+1).
Example 46
Preparation of Compound (47a)
##STR00229##
[0446] Preparation of (47-2): 47-2 was prepared from 46-1 (274 mg,
0.46 mmol) and propynoic anhydride (0.12 mL, 2 equiv.) in pyridine
(5 mL) in the same manner as described for 46-2 (260 mg, 80%).
[0447] Preparation of (47a): 47-2 (120 mg, 0.2 mmol) was treated
with 80% aq. HCOOH at R.T. for 3 hours. The mixture was evaporated,
coevaporated with toluene and purified on silica (10 g column) with
a CH.sub.2Cl.sub.2/MeOH solvent system (4-10% gradient) to yield
47a (62 mg, 75%). MS: m/z=404 (M-1).
Example 47
Preparation of Compound (48a)
##STR00230##
[0449] Preparation of (48-2): 48-2 was prepared from 46-1 (150 mg,
0.27 mmol) and valeric anhydride (0.11 mL, 2 equiv.) in pyridine (3
mL) in the same manner as described for 46-2 (150 mg, 73%).
[0450] Preparation of (48a): 48-2 (140 mg, 0.18 mmol) was treated
with 80% aq. HCOOH at R.T. for 3 hours. The mixture was evaporated
and purified on silica (10 g column) with a CH.sub.2Cl.sub.2/MeOH
solvent system (4-10% gradient) to yield 48a (70 mg, 84%). MS:
m/z=462 (M+1).
Example 48
Preparation of Compounds (49a), (50a) and (51a)
##STR00231##
[0452] Preparation of (49-2). (50-2) and (51-2): To a solution of
46-1 (1.26 g, 2.12 mmol) in pyridine (15 mL) were added n-octanoic
acid (0.34 mL, 1.0 equiv.), DCC (60% in xylene; 0.81 mL, 1 equiv.)
and DMAP (52 mg; 0.2 equiv.). The resulting mixture was stirred for
6 hours at R.T. The mixture was evaporated, and the residue
partitioned between CH.sub.2Cl.sub.2 (100 mL) and saturated aq.
NaHCO.sub.3 (25 mL). The organic layer was washed with water, brine
and dried (Na.sub.2SO.sub.4). The residue was treated with toluene.
The solid material was filtered off, and the filtrate was purified
on silica (25 g column) with a hexanes/EtOAc solvent system
(30-100% gradient) to yield 49-2 (0.57 g, 32%), 50-2 (0.18 g, 12%),
and 51-2 (0.2 g, 13%).
[0453] Preparation of (49a): A mixture of 49-2 (114 mg, 0.13 mmol)
and 80% aq. formic acid was stirred for 3 hours at R.T. The mixture
was evaporated and coevaporated with toluene and purified on silica
(10 g column) with a CH.sub.2Cl.sub.2/MeOH solvent system (2-8%
gradient) to yield 49a (53 mg, 75%). MS: m/z=544 (M-1).
[0454] Preparation of (50a): 50a (44 mg, 75% yield) was prepared
from 50-2 (104 mg, 0.14 mmol) in the same manner as described for
49a by using a 4-10% gradient of MeOH in CH.sub.2Cl.sub.2 for
purification. MS: m/z=418 (M-1).
[0455] Preparation of (51a): 51a (60 mg, 71% yield) was prepared
from 50-2 (140 mg, 0.2 mmol) in the same manner as described for
49a by using a 4-10% gradient of MeOH in CH.sub.2Cl.sub.2 for
purification. MS: m/z=418 (M-1).
Example 49
Preparation of Compound (52a)
##STR00232##
[0457] Preparation of (52-2): A solution of
N-(tert-butoxycarbonyl)-L-valine (0.41 g, 1.9 mmol) and
carbonyldiimidazole (0.31 g, 1.9 mmol) in THF (9 mL) was stirred at
R.T. for 1.5 hours. The mixture was then stirred at 40.degree. C.
for 20 mins. The mixture was added to a solution of 7a (0.42 g,
1.43 mmol) and DMAP (25 mg, 0.2 mmol) in DMF (8 mL) and TEA (4 mL)
at 80.degree. C. The reaction mixture was stirred at 80.degree. C.
for 1 h, then cooled and concentrated. The residue was partitioned
between tert-butyl methyl ether (100 mL) and water. The organic
layer was washed with water, brine and dried (Na.sub.2SO.sub.4).
The residue was purified on silica (25 g column) with a
CH.sub.2Cl.sub.2/MeOH solvent system (2-10% gradient) to yield 52-2
(0.32 g, 90% in the mixture with 5'-isomer), which was repurified
by RP-HPLC (10-100% B; A: water, B: MeOH). Yield: 0.25 g (35%).
[0458] Preparation of (52a): A solution of 52-2 (0.12 g; 0.24 mmol)
in EtOAc (0.6 mL) was treated with HCl/dioxane (4 M; 0.6 mL) for 20
mins. with vigorous shaking. The white precipitate was filtered,
washed with diethyl ether and dried to yield 52a as the
dihydrochloride salt (95 mg; 85%). MS: m/z=391 (M-1).
Example 50
Preparation of Compound (53a)
##STR00233##
[0460] Preparation of (53-2): To a solution of N-Boc-Val-OH (0.16
g, 0.74 mmol) and Et.sub.3N (0.14 mL, 1.0 mmol) in THF was added
53-1. The resulting mixture was evaporated, coevaporated with
pyridine and toluene and dissolved in THF (4 mL). DIPEA (0.38 mL,
2.2 mmol) was added, followed by BOP-Cl (0.28 g, 1.1 mmol) and
3-nitro-1,2,4-triazole (0.13 g, 1.1 mmol). The reaction mixture was
stirred at R.T. for 1 h. The mixture was diluted with
CH.sub.2Cl.sub.2 (40 mL) and washed with saturated aq. NaHCO.sub.3
and brine. The combined aqueous layers were back extracted with
CH.sub.2Cl.sub.2. The combined organic extract was dried
(Na.sub.2SO.sub.4), evaporated, and the residue was purified on
silica (10 g column) with a hexanes/0.5% Et.sub.3N/EtOAc solvent
system (20-100% gradient) to yield 53-2 (0.39 g, 81%).
[0461] Preparation of (53a): A mixture of 14-2 (0.37 g, 0.33 mmol)
and 80% aq. HCOOH (10 mL) was stirred at R.T. for 3 hours. The
mixture was evaporated, and the residue was partitioned between
water and CH.sub.2Cl.sub.2. The aqueous layer was washed with
CH.sub.2Cl.sub.2 and evaporated. The solid residue was suspended in
EtOAc (1.5 mL) and treated with 4N HCl in dioxane (1.5 mL) with
vigorous shaking. The solid was filtered, washed with diethyl ether
and purified by RP-HPLC (A: 0.5N HCOOH in water, B: 0.5 N HCOOH in
acetonitrile). The resulting formic acid salt of 5'-O-valyn ester
was converted into 53a dihydrochloride salt (63 mg, 40%) by
suspending in EtOAc (2 mL) and treatment with 4N HCl/dioxane (2
mL). MS: m/z=391 (M-1).
Example 51
Preparation of Compound (39a)
##STR00234##
[0463] Preparation of (39-2): A solution of 39-1 (1.3 g, 1.4 mmol)
in anhydrous MeOH (20 mL) was charged with Pd/C (1.3 g) and stirred
at 25.degree. C. under hydrogen (1 atm) atmosphere for 1 hour. The
solution was filtered, evaporated to dryness, and purified on a
silica gel column (DCM:MeOH=100:1 to 50:1) to give 39-2 (1.2 g,
92.3%) as a white solid.
[0464] Preparation of (39-3): To a solution of 39-2 (1.2 g, 1.3
mmol) in MeOH (40 mL) was added NH.sub.4F (370 mg, 10 mmol) at
25.degree. C. and stirred at 60.degree. C. for 6 hours. The
solution was filtered, evaporated to dryness, and purified on a
silica gel column (DCM:MeOH=200:1 to 20:1) to give 39-3 as a white
solid (249 mg, 30.7%). .sup.1H NMR (MeOD, 400 MHz) .delta. 7.92 (s,
1H), 7.19-7.33 (m, 12H), 6.83-6.85 (m, 2H), 5.50 (dd, J.sub.1=4.0
Hz, J.sub.2=14.8 Hz, 1H), 4.19-4.88 (m, 1H), 4.22 (dd, J.sub.1=5.2
Hz, J.sub.2=16.0 Hz, 1H), 3.76 (s, 3H), 3.41 (dd, J.sub.1=12.0 Hz,
J.sub.2=36.8 Hz, 2H), 1.52-1.74 (m, 2H), 0.87 (t, J=7.6 Hz, 3H);
ESI-LCMS: m/z 586.1 [M+H].sup.+.
[0465] Preparation of (39a): A solution of 39-3 of 80% formic
acid/20% water (3 mL) stood at RT for 2 hours, and then was
concentrated to dryness. The residue was co-evaporated with
MeOH/toluene (3 times) and then ethyl acetate added. The suspension
in ethyl acetate was heated at 70.degree. C. for 5 mins. The
solvent was removed using a pipet. This washing was repeated 3
times. The resulting product (44 mg) was further purified on
reverse-phase HPLC using acetonitrile/water as mobile phase to give
39a (20 mg) as an off-white solid. .sup.1H NMR (DMSO, 400 MHz)
.delta. 7.92 (s, 1H), 10.82 br, 1H), 7.96 (s, 1H), 6.56 (s, 2H),
5.99 (dd, J=6.0, 12.8 Hz, 1H), 5.65 (d, J=4.8 Hz, 1H), 5.58, 5.45
(2t, J=5.2 Hz, 0.5H, 0.5H), 5.25 (br, 1H), 4.19-4.88 (m, 1H), 4.22
(dd, J.sub.1=5.2 Hz, J.sub.2=16.0 Hz, 1H), 3.76 (s, 3H), 3.41 (dd,
J.sub.1=12.0 Hz, J.sub.2=36.8 Hz, 2H), 1.52-1.74 (m, 2H), 0.87 (t,
J=7.6 Hz, 3H); ESI-LCMS: m/z 443.6
[M+6-methyl-2-heptylamine)]+.
Example 52
Preparation of Compounds (55a) and (56a)
##STR00235##
[0467] 1,2,4-Triazol (21 mg, 0.3 mmol) was dissolved in the mixture
of CH.sub.3CN (0.7 mL) and Et.sub.3N (44 .mu.L, 0.31 mmol).
POCl.sub.3 (9 ul, 0.1 mmol) was added, and the mixture was kept at
R.T. for 20 mins. The white precipitate was filtered, and the
filtrate added to the dry nucleoside (28 mg, 0.05 mmol). The
reaction was controlled by TLC and monitored by the disappearance
of the starting nucleoside. After completion of the reaction,
tetrabutylammonium salt of pyrophosphate (150 mg) was added,
followed by DMF (0.5 mL) to get a homogeneous solution. After 1.5
hours at ambient temperature, the reaction was diluted with water
(4 mL) and extracted with DCM (2.times.5 mL). The combined organic
extracts were evaporated, dissolved in 5 mL of 80% HCOOH and left
for 2 hours at R.T. The reaction mixture was concentrated and
distributed between water (5 mL) and DCM (5 mL). The aqueous
fraction was loaded on the column HiLoad 16/10 with Q Sepharose
High Performance. Separation was done in a linear gradient of NaCl
from 0 to 1N in 50 mM TRIS-buffer (pH7.5). Two fractions were
obtained. The first fraction, containing the monophosphate (55a)
was eluted at 70-75% B. and triphosphate (56a) was eluted at 75-80%
B. Both fractions were desalted by RP HPLC on Synergy 4 micron
Hydro-RP column (Phenominex). A linear gradient of methanol from 0
to 30% in 50 mM triethylammonium acetate buffer (pH 7.5) was used
for elution. The corresponding fractions were combined,
concentrated and lyophilized 3 times to remove excess of
buffer.
Example 53
Preparation of Compounds (56b-e)
##STR00236##
[0469] 1,2,4-Triazol (21 mg, 0.3 mmol) was dissolved in the mixture
of CH.sub.3CN (0.7 mL) and Et.sub.3N (44 .mu.L, 0.31 mmol).
POCl.sub.3 (9 ul, 0.1 mmol) was added, and the mixture was kept at
R.T. for 20 mins. The white precipitate was filtered, and the
filtrate added to the dry nucleoside (28 mg, 0.05 mmol). The
reaction was controlled by TLC and monitored by the disappearance
of the starting nucleoside. After completion of the reaction,
tetrabutylammonium salt of pyrophosphate (150 mg) was added
followed by DMF (0.5 mL) to get a homogeneous solution. After 1.5
hours at ambient temperature, the reaction was diluted with water
(4 mL) and extracted with DCM (2.times.5 mL). The combined organic
extracts were evaporated, dissolved in 5 mL of 80% HCOOH and left
for 4 hours at 38.degree. C. The reaction mixture was concentrated
and distributed between water (5 mL) and DCM (5 mL). The aqueous
fraction was loaded on the column HiLoad 16/10 with Q Sepharose
High Performance. Separation was done in a linear gradient of NaCl
from 0 to 1N in 50 mM TRIS-buffer (pH7.5). Two fractions were
obtained. The triphosphate (56b-e) was eluted at 75-80% B. Desaltin
was performed by RP HPLC on Synergy 4 micron Hydro-RP column
(Phenominex). A linear gradient of methanol from 0 to 30% in 50 mM
triethylammonium acetate buffer (pH 7.5) was used for elution. The
corresponding fractions were combined, concentrated and lyophilized
3 times to remove excess of buffer.
TABLE-US-00003 TABLE 3 Triphosphates obtained from Example 53
Structure MS (M-1) P(.alpha.) P(.beta.) P(.gamma.) ##STR00237##
373.00 +3.64 (s) NA NA ##STR00238## 532.95 -6.67 -6.74 (d) -21.87
(t) -11.51 -11.63 (d) ##STR00239## 526.05 -6.33 -6.47 (d) -22.48
(t) -11.53 -11.64 (d) ##STR00240## 516.00 -63.2 (bs) -22.45 (t)
-11.64 (d) ##STR00241## 524.4 -10.57 -- 10.67 (d) -23.31 (t) -11.31
-11.94 (d) ##STR00242## 529.8 -6.17 (bs) -- 21.96 (bs) -- 11.42
(bs)
Example 54
Preparation of Compound (57a)
##STR00243##
[0471] 2'-Deoxy-2'-fluoro-4'-C-(ethenyl)guanosine (25a, 31 mg, 0.1
mmol) was dissolved in dry pyridine (3 mL). Isobutyric anhydrate
(50 .mu.L, 0.3 mmol) was added. The reaction mixture was kept at
ambient temperature. After 40 hours, isobutyric anhydrate (100
.mu.L, 0.6 mmol) was added, and the reaction mixture was left
overnight. The pyridine was evaporated. The residue was purified by
silica gel chromatography using a gradient of methanol in DCM from
3% to 10% to yield 57a (20 mg, 50%). .sup.1H NMR (DMSO-d6) .delta.:
10.72 (s, 1H), 7.88 (s, 1H), 6.47 (s, 2H), 6.18-6.13 (dd, 1H),
5.90-5.83 (dd, 1H), 5.79-5.62 (m, 2H), 5.49-5.44 (d, 1H), 5.35-5.32
(d, 1H), 4.28-4.25 (d, 1H), 4.12-4.10 (d, 1H), 2.60-2.45 (m, 2H),
1.12-1.09 (m, 6H), 1.02-0.96 (m, 6H); m/z 452 (M+1).
Example 55
Preparation of Compound (58a)
##STR00244## ##STR00245##
[0473] Preparation of (58-2): To a solution of 58-1 (50.0 g, 205
mmol) in pyridine (250 mL) was added DMTrCl (75.0 g, 225.0 mmol).
The solution was stirred at R.T. for 15 hours. MeOH (120 mL) was
added, and the mixture was concentrated to dryness under reduced
pressure. The residue was dissolved in EA and washed with water.
The organic layer was dried over Na.sub.2SO.sub.4 and concentrated
to give the crude DMTr protected derivative (80.5 g, 89%) as a
light yellow solid. Dried K.sub.2CO.sub.3 (80.52 g, 583.2 mmol) and
then PMBCl (31.7 g, 109.2 mmol) were added to a stirred solution of
the DMTr protected derivative (80 g, 146 mmol) in anhydrous DMF
(300 mL). The stirring was continued at ambient temperature for
overnight. The reaction was monitored by TLC. The mixture was
diluted with EA and washed with water. The organic layer was dried
over Na.sub.2SO.sub.4 and concentrated to give 58-2 (98.8 g, 90%)
as light yellow solid.
[0474] Preparation of (58-3): NaH (10.4 g, 260.5 mmol) and BnBr
(73.8 g, 434.2 mmol) were added to a stirred solution of 58-2 (98.8
g, 147.9 mmol) in anhydrous DMF (300 mL), and the stirring was
continued at 25.degree. C. overnight. The reaction was monitored by
TLC. The reaction was quenched with water, extracted with EA and
washed with brine. The solvent was removed, and the residue was
purified on silica gel (PE:EA=10:1 to 3:1) to give the Bn protected
derivative (101.1 g, 90%) as a light yellow solid. The Bn protected
derivative (101.1 g, 133.4 mmol) was dissolved in 80% HOAc (900 mL)
at 25.degree. C. The mixture was stirred at 25.degree. C.
overnight. The reaction was quenched with MeOH, and the solvent was
removed to give the alcohol (42.1 g, 70%) as a white foam. To a
solution of the alcohol (42.1 g, 92.6 mmol) in anhydrous CH.sub.3CN
(300 mL) was added IBX (28.5 g, 121.7 mmol) at 25.degree. C. The
reaction mixture was refluxed for 1 hour and then cooled to
0.degree. C. The precipitate was filtered-off, and the filtrate was
concentrated to give 58-3 (39.2 g, 93%) as a yellow solid.
[0475] Preparation of (58-4): To a solution of 58-3 (39.2 g, 86.39
mmol) in 1,4-dioxane (250 mL) was added 37% CH.sub.2O (28.1 mL,
345.6 mmol) and 2N NaOH aqueous solution (86.4 mL, 172.8 mmol). The
mixture was stirred at 25.degree. C. for 2 h and then neutralized
with AcOH to pH=7. To the reaction were added EtOH (200 mL) and
NaBH.sub.4 (19.7 g, 518.6 mmol). The mixture was stirred at
25.degree. C. for 30 mins. The reaction was quenched with saturated
aqueous NH.sub.4Cl. The mixture was extracted with EA, and the
organic layer was dried over Na.sub.2SO.sub.4 and concentrated. The
residue was purified by silica gel column chromatography (PE:EA=4:1
to 2:1) to give the diol derivative (25.5 g, 55%) as a white solid.
To a stirred solution of the diol derivative (25.5 g, 52.5 mmol) in
anhydrous pyridine (150 mL) and anhydrous CH.sub.3CN (150 mL) was
added BzCl (6.6 g, 52.47 mmol) dropwise at 0.degree. C. The mixture
was then stirred at 25.degree. C. for 14 h. The reaction was
quenched with H.sub.2O, and the solution was concentrated. The
residue was dissolved in EA and washed with NaHCO.sub.3. The
organic layer was dried over Na.sub.2SO.sub.4 and concentrated. The
residue was purified on a silica gel column (PE/EA=5:4) to give
58-4 (18.1 g, 60%) as a white foam.
[0476] Preparation of (58-5: Cs.sub.2CO.sub.3 (30.0 g, 92.0 mmol)
and BnBr (10.4 g, 61.3 mmol) were added to a stirred solution of
compound 58-4 (18.1 g, 30.6 mmol) in anhydrous DMF (300 mL), and
stirring was continued at 25.degree. C. overnight. The reaction was
quenched with NH.sub.4Cl, extracted with EA and washed with brine.
The solvent was removed to give the Bz protected derivative (19.3
g, 95%) as a light yellow solid. To a stirred solution of the Bz
protected derivative (19.3 g, 28.4 mmol) in anhydrous MeOH (230 mL)
was added NaOMe (24.9 g, 460 mmol) at 25.degree. C. for 1 h. The
reaction was quenched with AcOH (10 mL) and concentrated. The
residue was purified on a silica gel column (PE/EA=1/2) to afford
58-5 (11.2 g, 54%) as a white solid.
[0477] Preparation of (58-6): To a stirred solution of 58-5 (200
mg, 0.347 mmol) in anhydrous DCM (5 mL) was added DMP (168 mg,
0.674 mmol) at 25.degree. C. The mixture was stirred at 25.degree.
C. for 2 h. The solvent was removed, and the residue was purified
on a silica gel column (PE:EA=5:1 to 1:1) to give the aldehyde
derivative (161 mg, 81%). To a stirred solution of the aldehyde
derivative (200 mg, 0.348 mmol) in anhydrous THF (5 mL) was added
MeMgBr (1.0 mL, 1.01 mmol) at -78.degree. C. The mixture was
stirred at -78.degree. C. for 1 h. The reaction was quenched with
NH.sub.4Cl and extracted with EA. The concentrated organic phase
was purified by column chromatography (PE:EA=5:1 to 1:1) to give
58-6 (135 mg, 65%).
[0478] Preparation of (58-7): To a solution of 58-6 (900 mg, 1.5
mmol) in DCM was added DMP (2.5 g, 6.0 mmol) at 0.degree. C. After
stirring at 0.degree. C. for 1 h, the mixture was quenched with
Na.sub.2S.sub.2O.sub.3. The solvent was removed, and the residue
was purified on a silica gel column (PE:EA=5:1 to 1:1) to give the
ketone derivative (700 mg, 78%). To a solution of the ketone
derivative (700 mg, 1.52 mmol) in MeOH was added NaBH.sub.4 in
portions. After stirring at the same temperature for 1 h, the
mixture was quenched with water. The solvent was removed, and the
residue was purified on a silica gel column (PE:EA=5:1 to 1:1) to
give 58-7 (500 mg, 71%).
[0479] Preparation of (58-8): To a stirred solution of DAST (1.39
g, 8.68 mmol) in anhydrous toluene (15 mL) was added dropwise a
solution of 58-6 (1.0 g, 1.73 mmol) at -78.degree. C. The mixture
was stirred at -78.degree. C. for 30 min. The solution was warmed
to 25.degree. C. slowly and stirring continued overnight. The
mixture was poured into a saturated Na.sub.2CO.sub.3 solution. The
concentrated organic phase was purified on a silica gel column
(PE:EA=10:1 to 4:1) to give the fluoride derivative (449 mg, 45%).
A mixture of the fluoride derivative (1.20 g, 2.07 mmol) and CAN
(3.41 g, 6.23 mmol) in a 3:1 solution of MeCN and water (10 mL) was
stirred at 25.degree. C. overnight. Brine (10 mL) was added, and
the mixture extracted with EA. The combined organic extracts were
dried and evaporated under reduced pressure. Purification by
chromatography on silica with PE:EA=10:1 to 2:1 gave 58-8 as a
yellow solid (475 mg, 50%).
[0480] Preparation of (58-9): To a stirred solution of 58-8 (550
mg, 210 mmol) in anhydrous MeCN (10 mL) were added TPSCl (725 mg,
2.40 mmol), DMAP (293 mg, 2.40 mmol) and TEA (242 mg, 2.40 mmol) at
25.degree. C. The mixture was stirred at 25.degree. C. overnight.
NH.sub.4OH (25 mL) was added and stirred for 2 h. The solvent was
removed, and the residue was purified on a silica gel column
(DCM:MeOH=10:1) to give 58-9 (300 mg). .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 7.70 (d, J=8.4 Hz, 1H), 7.25-7.36 (m, 10H), 6.13 (dd,
J=2.8, 16.8 Hz, 1H), 5.40 (d, J=7.6 Hz, 1H), 5.15 (m, 1H), 4.81 (d,
J=11.6 Hz, 1H), 4.40-4.52 (m, 4H), 3.82 (d, J=8.8 Hz, 7H), 3.62 (d,
J=9.6 Hz, 7H), 1.35 (dd, J=2.8, 14.4 Hz, 3H). ESI-MS: m/z 472.1
[M+H].sup.+.
[0481] Preparation of (58a): A 1 M boron trichloride solution in
CH.sub.2Cl.sub.2 (3.2 mL; 3.2 mmol) was added dropwise to a
solution of 58-9 (200 mg, 0.42 mmol) in anhydrous CH.sub.2Cl.sub.2
(10 mL) at -78.degree. C. The mixture was slowly (in 4 h) warmed to
-30.degree. C. and stirred at -30 to -20.degree. C. for 3 h.
Ammonium acetate (1 g) and MeOH (5 mL) were added, and the
resulting mixture allowed to warm to ambient temperature. The
solvent was removed, and residue purified by RP-HPLC (0-60% B; A:
50 mM aqueous TEAA, B: 50 mM TEAA in MeOH) to yield 58a (75 mg).
.sup.1H NMR (CD.sub.3OD) .delta. 7.97 (d, 1H), 6.20 (dd, 1H), 5.92
(d, 1H), 5.22 (dt, 1H), 4.98 (dq, 1H), 4.58 (dd, 1H), 3.73 (m, 2H),
1.40 (dd, 3H). .sup.19F NMR (CD.sub.3OD) .delta. -205.80 (m, 1F),
-188.54 (m, 1F). ESI-MS: m/z 290.4 [M-H].sup.-.
Example 56
Preparation of Compound (59a)
##STR00246##
[0483] Preparation of (59-2): To a solution of 59-1 (100.0 g, 406.5
mmol) in pyridine (750 mL) was added DMTrCl (164.9 g, 487.8 mmol).
The solution was stirred at R.T. for 15 h. MeOH (300 mL) was added,
and the mixture was concentrated to dryness under reduced pressure.
The residue was dissolved in EtOAc and washed with water. The
organic layer was dried over Na.sub.2SO.sub.4 and concentrated. The
residue was dissolved in DCM (500 mL). To this solution were added
imidazole (44.3 g, 650.4 mmol) and TBSCl (91.9 g, 609.8 mmol). The
resulting reaction mixture was stirred at R.T. for 14 h. The
reaction solution was washed with NaHCO.sub.3 and brine. The
organic layer was dried over Na.sub.2SO.sub.4, and concentrated to
give the crude product as a light yellow solid. The crude product
(236.4 g, 356.6 mmol) was dissolved in 80% HOAc aqueous solution
(500 mL). The mixture was stirred at R.T. for 15 h. The mixture was
diluted with EtOAc, washed with NaHCO.sub.3 solution and brine. The
organic layer was dried over Na.sub.2SO.sub.4 and purified on a
silica gel column chromatography (1-2% MeOH in DCM) to give 59-2
(131.2 g, 89.6%) as a light yellow solid. .sup.1H NMR (DMSO-d6, 400
MHz) .delta. 11.39 (s, 1H), 7.88 (d, J=7.2 Hz, 1H), 5.89 (dd,
J=18.0 Hz, J=2.0 Hz, 1H), 5.64 (d, J=8.0 Hz, 1H), 5.21 (dd,
J.sub.1=J.sub.2=7.2 Hz, 1H), 5.18-5.03 (m, 1H), 4.37-4.29 (m, 1H),
3.86 (dd, J=3.2 Hz, J=3.2 Hz, 3H), 3.78-3.73 (m, 1H), 3.51-3.56 (m,
1H), 3.31 (s, 1H), 0.89 (s, 9H), 0.11 (s, 6H); ESI-MS: m/z 802
[M+H].sup.+.
[0484] Preparation of (59-3): To a solution of 59-2 (131.2 g, 364.0
mmol) in anhydrous CH.sub.3CN (1200 mL) was added IBX (121.2 g,
432.8 mmol) at R.T. The reaction mixture was refluxed for 3 h and
then cooled to 0.degree. C. The precipitate was filtered-off, and
the filtrate was concentrated to give the crude aldehyde (121.3 g)
as a yellow solid. The aldehyde was dissolved in 1,4-dioxane (1000
mL). 37% CH.sub.2O (81.1 mL, 1.3536 mol) and 2M NaOH aqueous
solution (253.8 mL, 507.6 mmol) were added. The mixture was stirred
at R.T. for 2 h and then neutralized with AcOH to pH=7. To the
solution were added EtOH (400 mL) and NaBH.sub.4 (51.2 g, 1.354
mol). The mixture was stirred at R.T. for 30 mins and quenched with
sat. aqueous NH.sub.4Cl. The mixture was extracted with EA. The
organic layer was dried over Na.sub.2SO.sub.4 and concentrated. The
residue was purified by silica gel column chromatography (1-3% MeOH
in DCM) to give 59-3 (51.4 g, 38.9%) as a white solid.
[0485] Preparation of (59-4): To a solution of 59-3 (51.4 g, 131.6
mmol) in anhydrous DCM (400 mL) were added pyridine (80 mL) and
DMTrCl (49.1 g, 144.7 mmol) at 0.degree. C. The reaction was
stirred at R.T. for 14 h, and then treated with MeOH (30 mL). The
solvent was removed, and the residue was purified by silica gel
column chromatography (1-3% MeOH in DCM) to give the mono-DMTr
protected intermediate as a yellow foam (57.4 g, 62.9%). To the
mono-DMTr protected intermediate (57.4 g, 82.8 mmol) in
CH.sub.2Cl.sub.2 (400 mL) was added imidazole (8.4 g, 124.2 mmol)
and TBDPSCl (34.1 g, 124.2 mmol). The mixture was stirred at R.T.
for 14 h. The precipitated was filtered off, and the filtrate was
washed with brine and dried over Na.sub.2SO.sub.4. The solvent was
removed to give the residue (72.45 g) as a white solid, which was
dissolved in 80% HOAc aqueous solution (400 mL). The mixture was
stirred at R.T. for 15 h. The mixture was diluted with EtOAc,
washed with NaHCO.sub.3 solution and brine. The organic layer was
dried over Na.sub.2SO.sub.4 and purified by silica gel column
chromatography (1-2% MeOH in DCM) to give 59-4 (37.6 g, 84.2%) as a
white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 7.76 (d,
J=4.0 Hz, 1H), 7.70 (dd, J=1.6 Hz, J=8.0 Hz, 2H), 7.66-7.64 (m,
2H), 7.48-7.37 (m, 6H), 6.12 (dd, J=2.8 Hz, J=16.8 Hz, 1H), 5.22
(d, J=8.0 Hz, 1H). 5.20-5.05 (m, 1H), 4.74 (dd, J=5.6 Hz, J=17.6
Hz, 1H), 4.16 (d, J=12.0 Hz, 1H), 3.87-3.80 (m, 2H), 3.56 (d,
J=12.0 Hz, 1H), 1.16 (s, 9H), 0.92 (s, 9H), 0.14 (s, 6H).
[0486] Preparation of (59-5): To a solution of 59-4 (3.0 g, 4.78
mmol) in anhydrous DCM (100 mL) was added Dess-Martin periodinane
(10.4 g, 23.9 mmol) at 0.degree. C. under nitrogen. The reaction
mixture was stirred at R.T. for 5 h. The mixture was poured into
NaHCO.sub.3 and Na.sub.2S.sub.2O.sub.3 (1:1) aqueous solution. The
organic layer was dried over anhydrous Na.sub.2SO.sub.4 and
concentrated to give a residue. The residue was purified on a
silica gel column (20% EtOAc in PE) to give the intermediate (2.5
g, 83.1%) as a white solid.
[0487] To a mixture of bromotriphenyl(propyl)phosphorane (6.45 g,
16.8 mmol) in anhydrous THF (3 mL) was added t-BuOK (16.8 mL, 16.8
mmol) at 0.degree. C. under nitrogen. The reaction mixture was
stirred at 0.degree. C. for 50 mins. A solution of the above
intermediate (1.5 g, 2.4 mmol) in anhydrous THF (3 mL) was added
dropwise at 0.degree. C. under nitrogen. The reaction mixture was
stirred at R.T. for 3 h. The reaction was quenched by NH.sub.4Cl
aqueous solution and extracted with EtOAc. The organic layer was
dried over anhydrous Na.sub.2SO.sub.4 and concentrated to give a
residue. The residue was purified on a silica gel column (20% EtOAc
in PE) to give 59-5 (1.3 g, 83%) as a white solid.
[0488] Preparation of (59a): To a solution of 59-5 (300 mg, 0.45
mmol) in anhydrous CH.sub.3CN (2 mL) were added TPSCl (341 mg, 1.13
mmol), DMAP (138 mg, 1.13 mmol) and NEt.sub.3 (571 mg, 5.65 mmol)
at R.T. The reaction mixture was stirred at R.T. for 2 h.
NH.sub.4OH (1 mL) was added, and the reaction mixture was stirred
for 1 h. The mixture was diluted with EA and washed with water. The
organic layer was dried and concentrated to give a residue. The
residue was purified on a silica gel column (2% MeOH in DCM) to
give the cytidine derivative (285 mg, 95.0%) as a white solid.
[0489] To a solution of the cytidine derivative (280 mg, 0.43 mmol)
in MeOH (10 mL) was added NH.sub.4F (1.0 g) at R.T. The reaction
mixture was refluxed for 12 h. The mixture was filtered, and the
filtrate was concentrated. The residue was purified on a silica gel
column (10% MeOH in DCM) to give 59a (81 mg, 61%) as a white solid.
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.11 (d, J=8.0 Hz, 1H),
5.91 (dd, J=1.2 Hz, J=17.6 Hz, 1H), 5.90 (d, J=7.6 Hz, 1H),
5.57-5.59 (m, 2H), 4.82-4.96 (m, 1H), 4.42 (dd, J=4.8 Hz, J=24.4
Hz, 1H), 3.72 (d, J=12.4 Hz, 1H) 3.58 (d, J=12.4 Hz, 1H), 2.31-2.41
(m, 2H), 0.99 (t, J=7.6 Hz, 3H). ESI-TOF-MS: m/z 300.1
[M+H].sup.+.
Example 57
Preparation of Compound (60a)
##STR00247##
[0491] Preparation of (60-1): To a solution of 59-5 (450 mg, 0.69
mmol) in MeOH (10 mL) was added Pd/C (200 mg) at R.T. The reaction
mixture was stirred R.T. for 1 h under H.sub.2 (balloon). The
mixture was filtered, and the filtrate was concentrated to give
crude 60-1 (440 mg, 97.1%) as a white solid.
[0492] Preparation of (60a): To a solution of 60-1 (440 mg, 0.67
mmol) in anhydrous CH.sub.3CN (2 mL) were added TPSCl (510 mg, 1.68
mmol), DMAP (205 mg, 1.68 mmol) and NEt.sub.3 (338 mg, 3.35 mmol)
at R.T. The reaction mixture was stirred at R.T. for 2 h.
NH.sub.4OH (1 mL) was added, and the reaction was stirred for 1 h.
The mixture was diluted with EA and washed with water. The solvent
was removed. The crude product was purified on a silica gel column
(2% MeOH in DCM) to give the cytidine derivative (205 mg, 46.5%) as
a white solid.
[0493] To a solution of the cytidine derivative (205 mg, 0.31 mmol)
in MeOH (6 mL) was added NH.sub.4F (0.6 g) at R.T. The reaction
mixture was refluxed overnight. After cooling to R.T., the mixture
was filtered. The filtrate was concentrated, and the residue was
purified on a silica gel column (10% MeOH in DCM) to give 60a (59
mg, 62.8%) as a white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 8.09 (d, J=7.6 Hz, 1H), 6.01 (dd, J=3.2 Hz, J=15.6 Hz, 1H),
5.89 (d, J=7.2 Hz, 1H), 4.95-5.12 (m, 1H), 4.41 (dd, J=5.2 Hz,
J=17.2 Hz, 1H), 3.75 (d, J=12.0 Hz, 1H) 3.56 (d, J=11.6 Hz, 1H),
1.73-1.80 (m, 1H), 1.55-1.63 (m, 1H), 1.40-1.46 (m, 4H), 0.92 (t,
J=7.6 Hz, 3H). ESI-MS: m/z 301.8 [M+H].sup.+.
Example 58
Preparation of Compound (61a)
##STR00248##
[0495] Preparation of (61-1): To a solution of 59-4 (1.5 g, 2.39
mmol) in anhydrous DCM (100 mL) was added Dess-Martin periodinane
(5.2 g, 11.95 mmol) at 0.degree. C. under nitrogen. The reaction
mixture was stirred at R.T. for 5 h. The mixture was poured into
NaHCO.sub.3 and Na.sub.2S.sub.2O.sub.3 solution and washed with
brine. The organic layer was dried with anhydrous Na.sub.2SO.sub.4,
and concentrated to give the crude intermediate (1.5 g) as a white
solid.
[0496] To a solution of the crude intermediate (1.5 g, 2.39 mmol)
in THF (12 mL) was added methylmagnesium bromide (2.4 mL, 7.2 mmol)
dropwise at 0.degree. C. The resulting mixture was stirred at
0.degree. C. for 2 h. After the starting material was consumed, the
reaction was quenched with saturated NH.sub.4Cl. The reaction
mixture was extracted with DCM. The organic layer was washed with
brine, dried and concentrated to give crude 61-1 (1.5 g).
[0497] Preparation of (61-2): To a solution of 61-1 (1.5 g, 2.39
mmol) in anhydrous DCM (50 mL) was added Dess-Martin periodinane
(4.5 g, 10.6 mmol). The reaction mixture was stirred at R.T.
overnight. The mixture was poured into NaHCO.sub.3 and
Na.sub.2S.sub.2O.sub.3 aqueous solution. The organic layer was
separated, washed with brine, dried and concentrated to give a
residue. The residue was purified on a silica gel column (10% EtOAc
in PE) to give the intermediate (907 mg, 58.6%) as a white
solid.
[0498] To a mixture of bromo(methyl)triphenylphosphorane (5.0 g, 14
mmol) in anhydrous THF (8 mL) was added t-BuOK (12.6 mL, 12.6 mmol)
at 0.degree. C. under nitrogen. The mixture was stirred at R.T. for
50 mins. A solution of the above intermediate (900 mg, 1.4 mmol) in
anhydrous THF (4 mL) was added dropwise at 0.degree. C. under
nitrogen. The reaction mixture was stirred at R.T. for 3 h. The
reaction mixture was quenched with NH.sub.4Cl aqueous solution and
extracted with DCM. The organic layer was separated, washed with
brine, dried and concentrated to give a residue. The residue was
purified on a silica gel column (5% EtOAc in PE) to give 61-2 (700
mg, 78.0%) as a white solid.
[0499] Preparation of (61a): To a solution of 61-2 (298 mg, 0.46
mmol) in anhydrous CH.sub.3CN (5.5 mL) were added TPSCl (346.5 mg,
1.14 mmol), DMAP (139.6 mg, 1.14 mmol) and NEt.sub.3 (115.6 mg,
1.14 mmol) at R.T. The reaction mixture was stirred at R.T. for 2
h. NH.sub.4OH (1 mL) was added, and the mixture was stirred for
another 1 h. The mixture was diluted with DCM and washed with
water. The organic layer was separated, washed with brine, dried
and concentrated to give a residue. The residue was purified on a
silica gel column (2% MeOH in DCM) to give the cytidine derivative
(250 mg, 85.0%) as a white solid.
[0500] To a solution of the cytidine derivative (250 mg, 0.39 mmol)
in MeOH (10 mL) was added NH.sub.4F (1.0 g) at R.T. The reaction
was refluxed for 12 h. The mixture was filtered, and the filtrate
was concentrated. The residue was purified on a silica gel column
(10% MeOH in DCM) to give 61a (55 mg, 49%) as a white solid.
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.11 (d, J=7.6 Hz, 1H),
6.21 (dd, J=4.2 Hz, J=14.0 Hz, 1H), 5.91 (d, J=7.6 Hz, 1H), 5.10
(dt, J=4.8 Hz, J=53.6 Hz, 1H), 5.13 (brs, 1H), 5.00 (brs, 1H), 4.46
(dd, J=4.8 Hz, J=11.6 Hz, 1H), 3.83 (d, J=11.6 Hz, 1H), 3.54 (d,
J=11.6 Hz, 1H), 1.84 (s, 3H). ESI-MS: m/z 285.9 [M+H].sup.+.
Example 59
Preparation of Compound (62a)
##STR00249##
[0502] Preparation of (62-1): To a solution of 61-2 (400 mg, 0.63
mmol) in MeOH (10 mL) was added Pd/C (400 mg) at R.T. The reaction
was stirred at R.T. for 5 h under H.sub.2 (balloon). The mixture
was filtered, and the filtrate was concentrated to give crude 62-2
(350 mg, 87%) as a white solid.
[0503] Preparation of (62a): To a solution of 62-1 (350 mg, 0.55
mmol) in anhydrous CH.sub.3CN (6 mL) were added TPSCl (414 mg, 1.4
mmol), DMAP (166.8 mg, 1.4 mmol) and NEt.sub.3 (138.1 mg, 1.4 mmol)
at R.T. The reaction mixture was stirred at R.T. for 2 h.
NH.sub.4OH (1 mL) was added, and the reaction was stirred for
another 1 h. The mixture was diluted with EA and washed with water.
The organic layer was separated, dried and concentrated to give a
residue. The residue was purified on a silica gel column (2% MeOH
in DCM) to give the cytidine derivative (300 mg, 85%) as a white
solid.
[0504] To a solution of the cytidine derivative (300 mg, 0.47 mmol)
in MeOH (10 mL) was added NH.sub.4F (1.5 g) at R.T. The reaction
mixture was refluxed overnight. After cooling to R.T., the mixture
was filtered. The filtrate was concentrated. The crude product was
purified on a silica gel column (10% MeOH in DCM) to give 62a (83
mg, 61%) as a white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 8.12 (d, J=7.6 Hz, 1H), 6.22 (dd, J=6.4 Hz, J=12.4 Hz, 1H),
5.94 (d, J=7.6 Hz, 1H), 5.25 (dt, J=5.6 Hz, J=54.0 Hz, 1H), 4.38
(t, J=4.8 Hz, 1H), 3.72 (d, J=11.6 Hz, 1H), 3.67 (d, J=11.6 Hz,
1H), 2.31-2.42 (m, 1H), 0.99 (2d, J=7.2 Hz, 6H). ESI-MS: m/z 287.8
[M+H].sup.+.
Example 60
Preparation of Compound (63a)
##STR00250## ##STR00251##
[0506] Preparation of (63-2): To a solution of 63-1 (50 g, 203
mmol) in anhydrous pyridine (200 mL) was added TBDPS-Cl (83.7 g,
304 mmol). The reaction was allowed to proceed overnight at R.T.
The solution was concentrated under reduced pressure to give a
residue. The residue was partitioned between ethyl acetate and
water. The organic layer was separated, washed with brine, dried
over magnesium sulfate and concentrated under reduced pressure to
give 5'-OTBDPS ether as a white foam (94 g).
[0507] To a solution of the 5'-OTBDPS ether (94.0 g, 194.2 mmol) in
anhydrous DCM (300 mL) were added silver nitrate (66.03 g, 388.4
mmol) and collidine (235 mL, 1.94 mol). The mixture was stirred at
R.T. After most of silver nitrate was dissolved (.about.15 min),
the mixture was cooled to 0.degree. C. Monomethoxytrityl chloride
(239.3 g, 776.8 mmol) was added as a single portion, and the
mixture was stirred overnight at R.T. The mixture was filtered
through Celite, and the filtrate was diluted with MTBE. The
solution was washed successively with 1M citric acid, diluted brine
and 5% sodium bicarbonate. The organic solution was dried over
sodium sulfate and concentrated under vacuum to give the fully
protected intermediate as a yellow foam.
[0508] The fully protected intermediate was dissolved in toluene
(100 mL), and the solution was concentrated under reduced pressure.
The residue was dissolved in anhydrous THF (250 mL) and treated
with TBAF (60 g, 233 mmol). The mixture was stirred for 2 hours at
R.T., and the solvent was removed under reduced pressure. The
residue was taken into ethyl acetate, and the solution was washed
with saturated sodium bicarbonate and brine. After drying over
magnesium sulfate, the solvent was removed in vacuum. The residue
was purified by column chromatography (PE:EA=5:1, 1:1) to give 63-2
(91 g, 86.4%) as a white foam.
[0509] Preparation of (63-3): To a solution of 63-2 (13.5 g, 26
mmol) in DCM (100 mL) was added pyridine (6.17 mL, 78 mmol). The
solution was cooled to 0.degree. C. and Dess-Martin periodinane
(33.8 g, 78 mmol) was added as a single portion. The reaction
mixture was stirred for 4 h at R.T. The reaction was quenched with
Na.sub.2S.sub.2O.sub.3 solution (4%) and sodium bicarbonate aqueous
solution (4%) (the solution was adjusted to pH 6, -150 mL). The
mixture was stirred for 15 min. The organic layer was separated,
washed with diluted brine and concentrated under reduced pressure.
The residue was dissolved in dioxane (100 mL), and the solution was
treated with 37% aqueous formaldehyde (21.2 g, 10 eq) and 2N
aqueous sodium hydroxide (10 eq). The reaction mixture was stirred
at R.T. overnight. After stirring for 0.5 h at R.T., the excess of
aqueous sodium hydroxide was neutralized with saturated with
NH.sub.4Cl (.about.150 mL). The mixture was concentrated under
reduced pressure. The residue was partitioned between ethyl acetate
and 5% sodium bicarbonate. The organic phase was separated, washed
with brine, dried over magnesium sulfate and concentrated. The
residue was purified by column chromatography (MeOH:DCM=100:1-50:1)
to give 63-3 (9.2 g, 83.6%) as a white foam.
[0510] Preparation of (63-4): 63-3 (23 g, 42.0 mmol) was
co-evaporated with toluene twice. The residue was dissolved in
anhydrous DCM (250 mL) and pyridine (20 mL). The solution was
cooled to -35.degree. C. Triflic anhydride (24.9 g, 88.1 mmol) was
added dropwise over 10 mins. The reaction was stirring for 40 min
at -35.degree. C. When TLC (PE:EA=2:1 and DCM:MeOH=15:1) showed
that the reaction was complete, the reaction was quenched with
water (50 mL) at 0.degree. C. The mixture was stirred 30 mins,
extracted with EA. The organic phase was dried over
Na.sub.2SO.sub.4 and filtered through a silica gel pad. The
filtrate was concentrated under reduced pressure. The residue was
purified by column chromatography (PE:EA=100:1-1:1) to give 63-4
(30.0 g, 88.3%) as a brown foam.
[0511] Preparation of (63-5): 63-4 (30 g, 36.9 mmol) was
co-evaporated twice with toluene. The resulting bis-triflate was
dissolved in anhydrous DMF (150 mL), cooled to 0.degree. C. and
treated with sodium hydride (60% in mineral oil; 1.5 g, 40.6 mmol,
1.1 eq). The reaction mixture was stirred at R.T. for 1 h until TLC
(DCM:MeOH=15:1) showed the disappearance of the bis-triflate and
formation of the 2,5'-anhydro intermediate. Lithium chloride (4.6
g, 110.7 mmol, 3 eq) was added, and the stirring was continued for
2 h. The mixture was taken into 100 mL of half saturated ammonium
chloride and ethyl acetate. The organic phase was separated, washed
with diluted brine and concentrated under reduced pressure to give
63-5.
[0512] Preparation of (63-6): 63-5 was dissolved in THF (150 mL),
and the solution was treated with 1N aqueous sodium hydroxide
(.about.41 mL, 40.1 mmol, 1.1 eq). The mixture was stirred at R.T.
for 1 h. The reaction was monitored by LCMS. The reaction was
diluted with half saturated sodium bicarbonate (.about.60 mL) and
extracted with ethyl acetate. The organic phase was dried
(magnesium sulfate) and concentrated under reduced pressure.
Purification of the residue by column chromatography
(DCM:MeOH=300:1-60:1) gave 63-6 (18.3 g, 87.6%) as a yellow
foam.
[0513] Preparation of (63-7): To a solution of 63-6 (18.3 g, 32.33
mmol) in anhydrous DCM (150 mL) was added TBS-Cl (17.7 g, 64.6
mmol) and imidazole (6.6 g, 97 mmol). The reaction was allowed to
proceed overnight at R.T. The reaction was diluted with water and
extracted with DCM. The organic layer was separated, washed with
brine, dried over Na.sub.2SO.sub.4 and concentrated. Purification
of the residue by column chromatography (DCM:MeOH=300:1-80:1) gave
63-7 (18.4 g, 83.7%) as a white foam.
[0514] Preparation of (63-8): A solution of 63-7 (18.4 g, 27.1
mmol), DMAP (6.6 g, 54.0 mmol) and TEA (5.4 g, 54.0 mmol) in MeCN
(450 mL) was treated with 2,4,6-triispropylbenzenesulfonyl chloride
(TPSCl, 16.3 g, 54.0 mmol). The mixture was stirred at R.T. for 3
h. NH.sub.3 H.sub.2O (70 mL) was added, and the mixture was stirred
for 2 h. The solution was evaporated under reduced pressure, and
the residue was purified on a silica gel column (DCM:MeOH=100:1 to
15:1) to give 63-8 (18.0 g) as a light yellow solid.
[0515] Preparation of (63-9): To a solution of 63-8 (18.0 g, 26.5
mmol) in anhydrous DCM (150 mL) was added collidine (8.1 g, 66.3
mmol, 2.5 eq), silver nitrate (4.5 g, 26.5 mmol, 1.0 eq) and DMTrCl
(13.4 g, 39.7 mmol, 1.5 eq). The reaction was allowed to proceed
overnight at R.T. The mixture was filtered. The filtrate was washed
with brine and extracted with DCM. The organic layer was separated,
dried over Na.sub.2SO.sub.4 and concentrated. The residue was
purified by column chromatography (PE:EA=60:1-3:1) as a yellow
foam. The foam was dissolved in THF (150 mL), and TBAF (10.4 g,
39.7 mmol, 1.5 eq) was added. The reaction was allowed to proceed
overnight at R.T. The mixture was concentrated, washed with brine
and extracted with EA. The organic layer was separated, dried over
Na.sub.2SO.sub.4 and concentrated. Purification of the residue by
column chromatography (PE:EA=60:I-EA) gave 63-9 (21.3 g, 92.4%) as
a yellow foam.
[0516] Preparation of (63-10): To a solution of 63-9 (2.0 g, 2.3
mmol) in anhydrous DCM (20 mL) was added Dess-Martin periodinane
(1.95 g, 4.6 mmol) at 0.degree. C. under nitrogen. The reaction was
stirred at R.T. for 5 h. The mixture was diluted with EtOAc (100
mL) and washed with a mixture of saturated aqueous
Na.sub.2S.sub.2O.sub.3 and saturated aqueous NaHCO.sub.3. The crude
product was purified by column chromatography on silica gel
(PE:EtOAc=2:1) to give 63-10 (1.8 g, 90%) as a yellow solid.
[0517] Preparation of (63-11): To a solution of tetramethyl
methylenediphosphonate (390 mg, 1.68 mmol) in anhydrous THF (10 mL)
was added NaH (84 mg, 2.1 mmol) at 0.degree. C. under nitrogen. The
reaction was stirred at 0.degree. C. for 30 min. A solution of
63-10 (1.2 g, 1.4 mmol) in anhydrous THF (10 mL) was added dropwise
at 0.degree. C. The reaction mixture was stirred at R.T. for 1 h.
The reaction was quenched by saturated aqueous NH.sub.4Cl, and the
crude product was purified by column chromatography on silica gel
(DCM:MeOH=150:1) to give 63-11 (1.2 g, 88.2%) as a yellow solid.
.sup.1H NMR (DMSO-d6, 400 M Hz) .delta. 8.51 (s, 1H), 7.46-7.09 (m,
22H), 6.88-6.82 (m, 6H), 6.62 (q, J.sub.1=17.2 Hz, J.sub.2=22.4 Hz,
1H), 6.12 (d, J=7.2 Hz, 1H), 5.86-5.75 (m, 2H), 5.43 (d, J=25.2 Hz,
1H), 4.63 (dd, J=4.8 Hz, J=21.2 Hz, 1H), 4.45 (d, J=12.0 Hz, 1H),
3.94 (d, J=12.0 Hz, 1H), 3.72 (s, 9H), 3.53 (q, J=11.2 Hz, J=16.0
Hz, 6H). ESI-MS: m/z 971.59 [M+H].sup.+.
[0518] Preparation of (63a): A solution of 63-11 (1.0 g, 1.03 mmol)
in 80% HOAc (46 mL) was stirred at 80-90.degree. C. for 2 h. The
solvent was removed, and the crude product was purified by column
chromatography on silica gel (DCM:MeOH=20:1) to give an
intermediate (337 mg, 82.3%) as a white solid. The intermediate was
dissolved in MeOH and wet Pd/C (300 mg) was added. The reaction
mixture was stirred under H.sub.2 (1 atm) for 1 h and then
filtered. The solvent was removed, and the residue was purified on
a silica gel column (DCM:MeOH=20:1) to give 63a (192 mg, 63.9%) as
a white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 7.60 (d,
J=7.6 Hz, 1H), 5.87 (d, J=7.2 Hz, 1H), 5.70 (dd, J=2.0 Hz, J=21.6
Hz, 1H), 5.31 (m, 1H), 4.67 (dd, J=5.6 Hz, J=19.6 Hz, 1H), 3.80 (m,
2H), 3.75 (2d, J=2.4 Hz, 6H), 1.92-2.20 (m, 4H). .sup.31P NMR
(CD.sub.3OD, 162 MHz) .delta. 35.77. ESI-MS: m/z 400.0
[M+H].sup.+.
Example 61
Preparation of Compound (64a)
##STR00252##
[0520] Preparation of (64-2): To a solution of 64-1 (1.0 g, 4.3
mmol) in THF (20 mL) was added NaH (120 mg, 3.0 mmol), and the
reaction mixture was stirred at 0.degree. C. for 1 h. Selectfluor
(1.2 g, 3.4 mmol) was added into the reaction mixture. The crude
product was purified on a silica gel column and eluted with EA to
give 64-2 (500 mg, 57%) as a white solid. .sup.1H NMR (CD.sub.3OD,
400 MHz) .delta. 5.65 (dt, J=14.0 Hz, J=44.8 Hz, 1H), 3.90 (d,
J=9.6 Hz, 12H).
[0521] Preparation of (64-3): To a solution of compound 64-2 (390
mg, 1.68 mmol) in anhydrous THF (10 mL) was added NaH (84 mg, 2.1
mmol) at 0.degree. C. under nitrogen. The reaction mixture was
stirred at 0.degree. C. for 30 mins. A solution of 63-10 (1.2 g,
1.4 mmol) in anhydrous THF (10 mL) was added dropwise at 0.degree.
C. The reaction mixture was stirred at R.T. for 1 h. The reaction
was quenched with saturated aqueous NH.sub.4Cl and concentrated to
give a residue. The residue was purified on a silica gel column
(DCM:MeOH=150:1) to give crude 64-3 (1.2 g, 88.2%) as a yellow
solid.
[0522] Preparation of (64a): A solution of crude 64-3 (230 mg, 0.23
mmol) in 80% HOAc (3 mL) was stirred at 80-90.degree. C. for 2 h.
The crude product was purified on a silica gel column (eluted with
DCM:MeOH=20:1) to give 64a (54 mg, 53.7%) as a white solid. H NMR
(DMSO, 400 MHz) .delta. 7.69 (d, J=7.2 Hz, 1H), 7.37 (d, J=1.6 Hz,
2H), 6.62-6.78 (m, 1H), 6.40 (d, J=5.6 Hz, 1H), 6.03-6.07 (m, 1H),
5.77 (d, J=7.6 Hz, 1H), 5.61-5.64 (m, 1H), 5.48-5.51 (m, 1H),
4.60-4.64 (m, 1H), 4.38 (d, J=11.6 Hz, 1H), 3.98 (d, J=11.6 Hz,
1H), 3.75 (2d, J=11.6 Hz, 6H). ESI-MS: m/z 416.3 [M+H].sup.+.
Example 62
Preparation of Compound (65a)
##STR00253##
[0524] A solution of crude 64-3 (230 mg, 0.23 mmol) in 80% HOAc (3
mL) was stirred at 80-90.degree. C. for 2 h. The crude product was
purified on a silica gel column (eluted with DCM:MeOH=20:1) to give
64a (52 mg, 33.7%) as a white solid. .sup.1H NMR (DMSO, 400 MHz)
.delta. 7.59 (d, J=7.2 Hz, 1H), 7.32 (s, 2H), 6.25-6.28 (m, 1H),
5.86-6.02 (m, 2H), 5.73 (s, 1H), 5.31 (d, J=14.0 Hz, 1H), 4.72 (d,
J=16.4 Hz, 1H), 3.90 (d, J=10.0 Hz, 1H), 3.73 (2d, J=11.6 Hz,
6H).
Example 63
Preparation of Compound (66a)
##STR00254##
[0526] A solution of 64a (130 mg, 0.3 mmol) in EA:MeOH (5:1, 20 mL)
was stirred under H.sub.2 (15 Psi) at R.T. for 2 h. The reaction
mixture was filtered and concentrated to give a residue. The
residue was purified on a silica gel column (DCM:MeOH=20:1) to give
66a (70 mg, 54%) as a white solid. .sup.1H NMR (DMSO, 400 MHz)
.delta. 7.61 (d, J=7.2 Hz, 1H), 5.87 (d, J=7.2 Hz, 1H), 5.58-5.80
(m, 1H), 5.26-5.47 (m, 2H), 4.97-5.03 (m, 1H), 5.58-5.80 (m, 1H),
3.73-3.94 (m, 6H), 2.33-2.59 (m, 2H). ESI-MS: m/z 418.3
[M+H].sup.+.
Example 64
Preparation of Compound (67a)
##STR00255##
[0528] Preparation of (67-2): To a solution of 67-1 (2.0 g, 6.9
mmol) in THF (20 mL) was added NaH (110 mg, 2.8 mmol), and the
reaction mixture was stirred at 0.degree. C. for 1 h. Selectfluor
(5.0 g, 13.6 mmol) was added into the reaction mixture. The
reaction was quenched with saturated NH.sub.4Cl and extracted with
EA. The organic layer was separated, dried and concentrated to give
the crude product. The crude product was purified on a silica gel
column (eluted with EA) to give 67-2 (600 mg, 28.3%) as a white
solid. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 5.65 (dt, J=14.0
Hz, J=44.8 Hz, 1H), 4.24-4.46 (m, 8H), 1.35-1.39 (m, 12H).
[0529] Preparation of (67-3): To a solution of 67-2 (2.14 g, 7.0
mmol) in anhydrous THF (10 mL) was added NaH (84 mg, 2.1 mmol) at
0.degree. C. under nitrogen. The reaction mixture was stirred at
0.degree. C. for 30 mins. A solution of 63-10 (3.0 g, 3.5 mmol) in
anhydrous THF (10 mL) was added in dropwise at 0.degree. C. The
reaction mixture was stirred at R.T. for 1 h. The reaction was
quenched with saturated aqueous NH.sub.4Cl and concentrated to give
a residue. The residue was purified on a silica gel column
(DCM:MeOH=150:1) to give crude 67-3 (2.9 g, 79.5%) as a yellow
solid.
[0530] Preparation of (67a): A solution of crude 67-3 (1.0 g, 0.98
mmol) in 80% HOAc (25 mL) was stirred at 80-90.degree. C. for 2 h.
The crude product was purified on a silica gel column (eluted with
DCM:MeOH=20:1) to give 67a (133 mg, 32.5%) as a white solid.
.sup.1H NMR (DMSO, 400 MHz) .delta. 7.67 (d, J=7.2 Hz, 1H), 7.34
(d, J=12.8 Hz, 2H), 6.33-6.69 (m, 1H), 6.05 (d, J=6.8 Hz, 1H),
6.00-6.05 (m, 1H), 5.76 (d, J=7.6 Hz, 1H), 5.45-5.61 (m, 1H),
4.60-4.63 (m, 1H), 4.08-4.14 (m, 5H), 1.23-1.29 (m, 6H). .sup.31P
NMR (DMSO, 162 MHz) .delta. 1.93, 1.30. ESI-MS: m/z 466.1
[M+Na].sup.+.
Example 65
Preparation of Compound (68a)
##STR00256##
[0532] To a solution of 67a (130 mg, 0.29 mmol) in MeOH (20 mL) was
stirred under H.sub.2 (15 Psi) at R.T. for 2 h. The reaction
mixture was filtered and concentrated to give a residue. The
residue was purified on a silica gel column (eluted with
DCM:MeOH=20:1) to give a mixture of diastereomers of 68a (90 mg,
69.2%) as a white solid. .sup.1H NMR (DMSO, 400 MHz) .delta.
7.61-7.68 (m, 1H), 7.28-7.38 (m, 2H), 5.89-5.95 (m, 1H), 5.58-5.79
(m, 2H), 5.18-5.39 (m, 2H), 4.53-4.85 (m, 1H), 4.04-4.39 (m, 4H),
3.71-3.83 (m, 2H), 2.21-2.35 (m, 2H), 1.21-1.27 (m, 6H). .sup.31P
NMR (DMSO, 162 MHz) .delta. 18.2, 18.02, 17.73, 17.56. ESI-MS: m/z
446.1 [M+H].sup.+
Example 66
Preparation of Compound (69a)
##STR00257##
[0534] Preparation of (69-1): 63-4 (3.0 g, 3.69 mmol) was
co-evaporated twice with toluene. The resulting bis-triflate was
dissolved in anhydrous DMF (20 mL). The solution was cooled to
0.degree. C. and treated with sodium hydride (60% in mineral oil;
177 mg, 0.43 mmol). The reaction was stirred at R.T. for 1 h (TLC
(PE:EA=2:1) showed complete disappearance of the bis-triflate and
clean formation of the 2',5'-anhydro intermediate). The reaction
mixture was used for the next step without any further workup
[0535] Preparation of (69-2): To the above stirred reaction mixture
was added NaSMe (9.0 g, 0.13 mmol) and 15-Crown-5 (4.87 g, 22.14
mmol) at 0.degree. C. under nitrogen. The solution was stirred at
R.T. for 2 h (TLC (PE:EA=1:1) showed the reaction was complete).
The reaction was quenched with water. The mixture was extracted by
EtOAc, washed with brine, and dried over MgSO.sub.4. The mixture
was filtered and concentrated to give a residue. The residue was
purified on a silica gel column (PE:EA=5:2) to give 69-2 (1.23 g,
59.0%) as a white foam.
[0536] Preparation of (69-3): To a stirred solution of 69-2 (1.34
g, 2.32 mmol) in anhydrous DCM (10 mL) was added MMTrCl (1.32 g,
4.64 mmol), AgNO3 (1.17 g, 6.96 mmol) and Collidine (1.41 g, 11.6
mmol) at R.T. under nitrogen. The reaction mixture was stirred at
R.T. for 1 h (TLC (PE:EA=1:1) showed the reaction was complete).
The mixture was filtered and concentrated. The residue was purified
on a silica gel column (PE:EA=8:1) to give 69-3 (1.31 g, 66.5%) as
a white foam.
[0537] Preparation of (69-4): To a solution of 69-3 (900 mg, 1.06
mmol) in anhydrous MeCN (9 mL) was added DMAP (259 mg, 2.12 mmol),
TEA (214 mg, 2.12 mmol) and TPSCl (640 mg, 2.12 mmol) at R.T. under
nitrogen. The reaction mixture was stirred at R.T. for 2 h (TLC
(DCM:MeOH=10:1) showed the reaction was complete). NH.sub.4OH (10
mL) was added, and the reaction mixture was stirred for another 1 h
(LCMS showed the reaction was complete). The solution was diluted
with water, extracted with EtOAc. The organic layer was washed with
1M HCl, saturated NaHCO.sub.3 and brine, and dried over MgSO.sub.4.
The mixture was filtered and concentrated to give a residue. The
residue was purified on a silica gel column (DCM:MeOH=70:1) to give
69-4 (870 mg, 68.5%) as a white solid.
[0538] Preparation of (69a): 69-4 (800 mg, 0.95 mmol) was dissolved
in 80% HOAc aq. (50 mL). The reaction mixture was heated to
75.degree. C. overnight (LCMS showed the reaction was complete).
The reaction mixture was concentrated and purified on a silica gel
column (DCM:MeOH=15:1) to give 69a (180 mg, 62.5%) as a white
solid. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.05 (d, J=7.2 Hz,
1H), 6.11 (dd, J=3.2 Hz J=15.6 Hz, 1H), 5.87 (d, J=7.6 Hz, 1H),
5.05 (dt, J=4.8 Hz, J=53.6 Hz, 1H), 4.47 (dd, J=5.2 Hz J=17.6 Hz,
1H), 3.83 (d, J=12.0 Hz, 2H), 2.84 (d, J=14.4 Hz, 2H), 2.15 (s,
3H). ESI-MS: m/z 305.8 [M+H].sup.+
Example 67
Preparation of Compound (70a)
##STR00258##
[0540] To a solution of 63-5 (100 g, 182.5 mmol) in MeCN (2 L) was
added 6N HCl aq. (15 g). The mixture was stirred at 40.degree. C.
for 7 h, and then neutralized to pH=5-6 with a 25% ammonia solution
(.about.8 g). The mixture was filtered to give a solid, which was
further washed by PE to give an intermediate (32.2 g, 60%) as a
white solid. To a mixture of the intermediate (32.2 g, 109.5 mmol),
TEA (22.1 g, 219 mmol) and DMAP (1.34 g, 11 mmol) in MeCN (1 L) was
added with isobutyric anhydrous (69.2 g, 438 mmol). The mixture was
stirred at R.T. for 3 h. The reaction was quenched by the addition
of water (200 mL) and extracted with 2-Me-THF (800 mL). The organic
layer was washed with saturated NaHCO.sub.3 and brine. The organic
layer was dried and concentrated to give a residue, which was
purified by a silica gel column (10% toluene in heptane) to give
70a (42.3 g, 89%) as a white solid. H NMR (CD.sub.3OD, 400 MHz)
.delta. 7.65 (d, J=8.0 Hz, 1H), 5.95 (dd, J=2.8, 20.4 Hz, 1H),
5.55-5.74 (m, 3H), 4.33-4.41 (m, 2H), 3.88 (s, 2H), 2.57-2.72 (m,
2H), 1.14-1.22 (m, 12H).
Example 68
Preparation of Compound (71a)
##STR00259##
[0542] Preparation of (71-1): To a solution of 63-4 (4.2 g, 5.17
mmol) in DMF (50 mL) at 0.degree. C. was added NaH (227 mg of 60%
dispersion, 5.7 mmol). The mixture was stirred at 0.degree. C. for
2 h, and then LiBr (1.34 g, 15.5 mmol) was added. The mixture was
stirred overnight at R.T., diluted with EA (150 mL) and washed
successively with water and brine. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified on a
silica gel column eluted with 10% EA in PE to give 71-1 as a yellow
solid (2 g, 66%)
[0543] Preparation of (71-2): To a solution of 71-1 (1.74 g, 2.9
mmol) in THF (20 mL) at 0.degree. C. was added 1N NaOH (3.2 mL, 3.2
mmol), and the mixture was stirred at 0.degree. C. for 2 h. The
mixture was partitioned between EA (100 mL) and water (20 mL), and
the organic layer was dried over Na.sub.2SO.sub.4 and evaporated to
dryness. The residue was purified on a silica gel column eluted
with 20% EA in PE to give the 5'-OH derivative as a yellow solid
(1.6 g, 90%).
[0544] To a solution of 5'-OH derivative (2.3 g, 3.76 mmol) in
anhydrous DCM (20 mL) were added collidine (0.8 g, 6.7 mol) and
MMTrCl (2.7 g, 8.7 mmol). The reaction mixture was stirred at R.T.
overnight. The mixture was filtered and washed successively with
saturated aqueous NaHCO.sub.3 and brine, dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified on a
silica gel column eluted with 10% EA in PE to give 71-2 as a yellow
solid (2.4 g, 73%).
[0545] Preparation of (71a): To a solution of 71-2 (2.4 g, 2.72
mmol) in anhydrous CH.sub.3CN (30 mL) were added TPSCl (1.65 g,
5.44 mmol), DMAP (0.663 g, 5.44 mmol) and NEt.sub.3 (1.5 mL) at
R.T. The mixture was stirred at R.T. for 3 h, and 28% aqueous
ammonia (30 mL) was added. The mixture was stirred for 1 h. The
mixture was diluted with EA (150 mL) and washed successively with
water, saturated aqueous NaHCO.sub.3 and brine. The solvent was
removed, and the residue was purified on a silica gel column eluted
with 2% MeOH in DCM to give a cytidine derivative as a yellow solid
(1.5 g, 62%).
[0546] The cytidine derivative (1.35 g, 1.5 mmol) was dissolved in
80% AcOH (40 mL), and the mixture was stirred at 60.degree. C. for
2 h. The mixture was concentrated, and the residue was purified on
a silica gel column using 5% MeOH in DCM as elute to give 71a as a
white solid (180 mg, 35%). .sup.1H NMR (MeOD, 400 MHz) .delta. 8.00
(d, J=7.2 Hz, 1H), 6.12 (dd, J=3.6 Hz, J=15.6 Hz, 1H), 5.88 (d,
J=7.6 Hz, 1H), 5.10 (dd, J=4.8 Hz, J=53.2 Hz, 1H), 4.59 (dd, J=5.2
Hz, J=16.4 Hz, 1H), 3.95 (d, J=11.6 Hz, 1H), 3.76 (d, J=11.6 Hz,
1H), 3.70 (d, J=11.6 Hz, 1H), 3.63 (d, J=11.2 Hz, 1H); ESI-TOF-MS:
m/z 337.9 [M+H].sup.+.
Example 69
Preparation of Compound (72a)
##STR00260##
[0548] Preparation of (72-1): To a solution of 63-6 (1.0 g, 1.8
mmol) in 1, 4-dioxane (2 mL) was added TEA (3 mL) and 37% HCHO (3
mL). The reaction mixture was stirred for 10 h at 60.degree. C. The
reaction was concentrated to dryness under vacuum, and the residue
was purified by column on a silica gel column (DCM:MeOH=100:1-30:1)
to give 72-1 (470 mg, 45%) as a white foam. .sup.1H NMR (DMSO-d6,
400 MHz) .delta. 11.4 (s, 1H), 7.27-7.49 (m, 13H), 6.89 (d, J=8.8
Hz, 2H), 4.90-4.95 (m, 1H), 4.58 (dd, J=5.2 Hz, J=23.6 Hz, 1H),
3.96-4.07 (m, 4H), 3.73 (s, 3H), 3.50-3.62 (m, 1H), 3.37-3.39 (m,
1H), ESI-TOF-MS: m/z 596.9 [M+H].sup.+.
[0549] Preparation of (72-2): To a solution of 72-1 (430 mg, 0.72
mmol) in dioxane (2 mL) was added 30% CH.sub.3COOH (0.7 mL) and
PtO.sub.2 (290 mg). The reaction mixture was stirred under H.sub.2
(1 atm) at R.T. for 2 h. The mixture was filtered, and the filtrate
was concentrated to dryness. The residue was purified on a silica
gel column (DCM:MeOH=100:1-30:1) to give 72-2 (268 mg, 64%) as a
white foam. .sup.1H NMR (DMSO-d6, 400 MHz) .delta. 11.3 (s, 1H),
7.27-7.46 (m, 13H), 6.88 (d, J=8.8 Hz, 2H), 5.78 (d, J=20.8 Hz,
1H), 5.06-5.08 (t, J=20.8 Hz, 1H), 4.49 (dd, J=4.2 Hz, J=24.4 Hz,
1H), 3.94-4.04 (m, 2H), 3.70 (s, 3H), 3.59-3.63 (m, 1H), 3.52-3.53
(m, 1H), 3.34-3.40 (m, 1H), 1.66 (s, 3H). ESI-TOF-MS: m/z 580.9
[M+H].sup.+.
[0550] Preparation of (72-3): To a solution of 72-2 (260 mg, 0.45
mmol) in anhydrous DCM (3 mL) was added AgNO.sub.3 (228 mg, 1.35
mmol), collidine (223 mg, 1.8 mmol) and MMTrC (456 mg, 1.35 mmol).
The mixture was stirred at R.T. for 10 h. The reaction mixture was
filtered, and the filtrate was concentrated to dryness. The residue
was purified on a silica gel column (PE:EA=50:1-3:1) to give 72-3
(303 mg, 80%) as a white foam.
[0551] Preparation of (72-4): To a solution of 72-3 (300 mg, 0.35
mmol) in anhydrous CH.sub.3CN (3 mL) was added DMAP (107 mg, 0.88
mmol), TEA (141 mg, 1.4 mmol) and TPSCl (106 mg, 0.35 mmol) at R.T.
The reaction mixture was stirred at R.T. for 4 h. NH.sub.4OH (1 mL)
was added, and the mixture was stirred at R.T. for another 1 h. The
solvent was removed, and the residue was partitioned by EA and
water. The organic layer was washed by brine twice, dried and
concentrated to give a residue. The residue was purified on a
silica gel column (PE:EA=50:1-3:1) to give 72-4 (270 mg, 90%) as a
white foam.
[0552] Preparation of (72a): 72-4 (260 mg, 0.31 mmol) in 10 mL of
60% HCOOH was stirred at R.T. for 2 h. The solvent was removed, and
the residue was washed with EA to give 72a (31 mg, 32%) as a white
powder. .sup.1H NMR (MeOD, 400 MHz) .delta. 7.85 (d, J=0.8 Hz, 1H),
6.12 (dd, J=4.0 Hz, J=15.2 Hz, 1H), 5.08-5.22 (m, 1H), 4.58 (dd,
J=4.8 Hz, J=14.8 Hz, 1H), 3.92 (d, J=15.6 Hz, 1H), 3.74-3.84 (m,
3H), 1.94 (d, J=0.8 Hz, 1H). ESI-TOF-MS: m/z 307.9 [M+H].sup.+.
Example 70
Preparation of Compound (73a)
##STR00261##
[0554] Preparation of (73-1): 63-6 (600 mg, 1.06 mmol) in formic
acid (5 mL, 80% in water) was stirred at R.T. overnight. Completion
of the reaction was determined by TLC (DCM:MeOH=10:1). The solvent
was removed to give crude 73-1 (290 mg, 93.2%).
[0555] Preparation of (73-2): To a solution of 73-1 (290 mg, 0.98
mmol) in pyridine (5 mL) and acetonitrile (5 mL) was added BzCl
(371 mg, 2.65 mmol). The reaction mixture was stirred at 0.degree.
C. for 0.5 h. The reaction was warmed to R.T. and stirred for 2 h.
Completion of the reaction was determined by LCMS. The reaction was
quenched with water and extracted with EA. The organic layer was
washed with brine, dried over MgSO.sub.4, filtered and
concentrated. The residue was purified on a silica gel column
(DCM:MeOH=200:1) to give 73-2 (245 mg, 49.8%) as a white solid.
[0556] Preparation of (73-3): To a solution of 73-2 (245 mg, 0.49
mmol) in anhydrous acetonitrile (2.5 mL) was added TPSCl (394 mg,
0.98 mmol), DMAP (119.5 mg, 0.98 mmol) and TEA (98 mg, 0.98 mmol).
The mixture was stirred at R.T. for 3 h. NH.sub.2OH HCl (68 mg,
0.98 mmol) and DBU (368 mg, 1.47 mmol) were added, and the reaction
mixture was stirred at R.T. for 2 h. The reaction mixture was
diluted with water and extracted with EtOAc. The combined organic
layer was washed with 1M HCl, saturated NaHCO.sub.3 and brine,
dried and concentrated. The residue was purified on a silica gel
column (DCM:MeOH=20:1) to give 73-3 (49 mg, 32.9%) as a white
solid.
[0557] Preparation of (73a): 73-3 (49 mg, 0.1 mmol) in
NH.sub.3/MeOH (30 mL) was stirred at R.T. for 2 days. The solvent
was removed. The residue was purified on a silica gel column
(DCM:MeOH=30:1) to give 73a (12.9 mg, 44.0%) as a white solid.
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 10.07 (brs, 1H), 9.68
(brs, 1H), 7.02 (d, J=8.0 Hz, 1H), 6.06 (dd, J=6.4 Hz, J=13.6 Hz,
1H), 5.94 (d, J=5.6 Hz, 1H), 5.60 (d, J=8.4 Hz, 1H), 5.36 (t, J=5.2
Hz, 1H), 5.16 (dt, J=5.2 Hz, J=53.6 Hz, H), 4.31-4.35 (m, 1H),
3.58-3.76 (m, 2H), 3.57-3.58 (m, 2H). ESI-TOF-MS: m/z 308.1
[M-H].sup.+.
Example 71
Preparation of Compound (74a)
##STR00262##
[0559] Preparation of (74-1): To a solution of 63-6 (1.2 g, 2.12
mmol) in anhydrous DCM (20 mL) were added collidine (750 mg, 6.51
mol) and MMTrCl (2.6 g, 8.5 mmol). The reaction mixture was stirred
at R.T. overnight. The reaction was filtered and washed
successively with saturated aqueous NaHCO.sub.3 and brine, dried
over Na.sub.2SO.sub.4 and concentrated. The residue was purified on
a silica gel column eluted with 10% EA in PE to give 74-1 as a
yellow solid (1.4 g, 72%).
[0560] Preparation of (74-2): To a stirred solution of 74-1 (600
mg, 0.715 mmol) in anhydrous acetonitrile (6 mL) were added TPSCl
(432 mg, 1.43 mmol), DMAP (174 mg, 1.43 mmol) and TEA (144 mg, 1.43
mmol). The mixture was stirred at R.T. for 2 h. Completion of the
reaction was determined by TLC (DCM:MeOH=10:1). CH.sub.3NH.sub.2
(310 mg, 10 mmol) was added dropwise at 0.degree. C. The reaction
mixture was stirred at R.T. for 2 h. The mixture was diluted with
water and extracted with EtOAc. The combined organic layer was
washed with 1M HCl, saturated NaHCO.sub.3 and brine. The solvent
was removed, and the residue was purified by prep-TLC
(DCM:MeOH=10:1) to give 74-2 (307 mg, 50.45%) as a white solid.
[0561] Preparation of (74a): 74-2 (300 mg, 0.352 mmol) in formic
acid (10 mL, 80% in water) was stirred at R.T. overnight.
Completion of the reaction was determined by TLC (DCM:MeOH=10:1).
The solvent was removed to dryness. The residue was dissolved in 20
mL of methanol. Ammonia (0.5 mL) was added, and the mixture was
stirred at R.T. for 5 mins. The solvent was removed, and the
residue was washed with PE (5.times.) to give 74a (103 mg, 95.3%)
as a white solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 7.79
(d, J=4.8 Hz, 1H), 7.72 (d, J=5.2 Hz, 1H), 6.10 (dd, J=4.4 Hz,
J=14.8 Hz, 1H), 5.97 (brs, 1H), 5.73 (d, J=7.6 Hz, 1H), 5.39 (brs,
1H), 5.08 (dt, J=4.2 Hz, J=53.2 Hz, 1H), 4.37-4.40 (m, 1H), 3.73
(s, 2H), 3.54-3.70 (m, 2H), 2.73 (d, J=4.4 Hz, 3H). ESI-TOF-MS: m/z
308.1 [M+H].sup.+.
Example 72
Preparation of Compound (75a)
##STR00263##
[0563] Preparation of (75-3): To a stirred solution of 75-1 (20.0
g, 151 mmol) in anhydrous THF (200 mL) was added NaH (7.8 g, 196
mmol) in portions at 0.degree. C. The mixture was stirred for 1 h,
and 75-2 (65.0 g, 196 mmol) was added dropwise at 0.degree. C. The
mixture was stirred at R.T. for 10 h. The reaction was quenched
with water and extracted with EA. The reaction was washed with
brine, and the organic layer was concentrated to obtain crude 75-3
(72 g).
[0564] Preparation of (75-4): Crude 75-3 (72 g, 151 mmol) was
dissolved with 80% CH.sub.3COOH (300 mL) and stirred for 10 h. The
solvent was removed under reduced pressure. The residue was
dissolved in EA and washed with saturated NaHCO.sub.3 and brine
successively. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated to dryness. The residue was purified on a silica gel
column to give the crude intermediate, which was dissolved in
anhydrous pyridine (80 mL) and DCM (400 mL). A solution of DMTrCl
(56.0 g, 166 mmol) in DCM (150 mL) was added dropwise at 0.degree.
C. The mixture was stirred at R.T. for 10 h. The reaction mixture
was concentrated to dryness, and the residue was purified by column
on silica gel (PE:EA=2:1) to give 75-4 (58.5 g, 61%).
[0565] Preparation of (75-5): To a stirred solution of 75-4 (10.0
g, 15.5 mmol) in anhydrous DMF (80 mL) was added NaH (0.8 g, 20
mmol) at 0.degree. C. The mixture was stirred at R.T. for 1 h, and
BnBr (33.8 g, 20 mmol) was added. The reaction mixture was stirred
at R.T. for 10 h. The reaction was quenched with water and
extracted with EA. The reaction was washed with brine, and the
organic layer was concentrated to give the crude intermediate (10.5
g, 92%) as a white foam. The crude intermediate (10.2 g, 13.8 mmol)
in 80% CH.sub.3COOH (100 mL) was stirred at R.T. for 12 h. The
solvent was removed. The residue was dissolved in EA, washed with
saturated NaHCO.sub.3 and brine successively, dried and
concentrated to give a residue. The residue was purified on a
silica gel column twice (PE:EA=3:1) to give 75-5 (4.2 g, 70%) as a
white foam.
[0566] Preparation of (75-6): To a solution of 75-5 (4.0 g, 9.2
mmol) in anhydrous CH.sub.3CN (30 mL) was added DIPEA (6.1 g, 47.6
mmol) and 2-cyanoethyl N,N-diisopropylchlorophosphoramidite (2.8 g,
11.9 mmol). The mixture was stirred at R.T. for 2 h. The solvent
was removed, and residue was partitioned by EA and saturated
NaHCO.sub.3. The organic layer was dried over MgSO.sub.4 and
concentrated to give a residue. The residue was purified on a
silica gel column (PE:EA=3:1) to give 75-6 (5.1 g, 88%) as a white
solid.
[0567] Preparation of (75-7): To a solution of 75-6 (1.0 g, 1.6
mmol) and 63-9 (925 mg, 1.1 mmol) in anhydrous MeCN (1 mL) was
added tetrazole (12 mL, 0.45M in MeCN, 5.5 mmol) dropwise at R.T.
After stirred for 3 h, TBDPH (0.96 mL, 5M 4.8 mmol) was added. The
reaction mixture was stirred at R.T. for 1 h. The mixture was
diluted with EA and washed with saturated Na.sub.2SO.sub.3 and
brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The
residue was purified by silica gel chromatography (PE/EA=50:1 to
1:1) to give 75-7 (1.1 g, 73.3%) as a white solid.
[0568] Preparation of (75a): 75-7 (1.0 g, 0.7 mmol) in 60% HCOOH (3
mL) was stirred at R.T. for 12 h. The solvent was removed. The
residue was dissolved in EA and washed with saturated NaHCO.sub.3
and brine successively, dried and concentrated to give a residue.
The residue was purified twice on a silica gel column
(DCM:MeOH=30:1) to give crude 75a (510 mg, 86%) as a white foam. To
a solution of crude 75a (275 mg, 0.33 mmol) in C.sub.2H5H was added
a few drops 1N NaOH until pH.about.7.0. The mixture was stirred for
0.5 h. The mixture was concentrated to give a residue. The residue
was purified by HPLC (MeCN and water, neutral system) to give 75a
(sodium salt, 170 mg, 64%) as a white solid. H NMR (CD.sub.3OD, 400
MHz) .delta. 8.01 (d, J=7.6 Hz, 1H), 7.23-7.37 (m, 5H), 6.22 (dd,
J=3.6 Hz, J=14.4 Hz, 1H), 6.01 (d, J=7.6 Hz, H), 5.01-5.16 (m, 1H),
4.63-4.72 (m, 2H), 4.52-4.11 (m, 1H), 4.23-4.29 (m, 1H), 3.91-4.09
(m, 3H), 3.69-3.81 (m, 3H), 3.51-3.60 (m, 2H), 3.41-3.45 (m, 2H),
1.48-1.55 (m, 2H), 1.21-1.35 (m, 32H), 0.87-0.91 (m, 3H). .sup.31P
NMR (CD.sub.3OD, 162 MHz) .delta. -0.223. ESI-TOF-MS: m/z 788.3
[M-H].sup.+.
Example 73
Preparation of Compound (76a)
##STR00264##
[0570] Preparation of (76-1): To a solution of 73-1 (4.1 g, 13.95
mmol) in pyridine (40 mL) was added Ac.sub.2O (3.13 g, 30.68 mmol)
at R.T., and the mixture was stirred overnight. The mixture was
concentrated, and the residue was purified on a silica gel column
(PE:EA=3:1) to give 76-1 (4.0 g, 75.9%).
[0571] Preparation of (76-2): To a solution of 76-1 (1.3 g, 3.44
mmol) in pyridine (20 mL) was added NBS (1.22 g, 6.88 mmol) at
R.T., and the mixture was stirred overnight. The mixture was
concentrated, and the residue was purified on a silica gel column
(PE:EA=4:1) to give 76-2 (1.43 g, 72.2%).
[0572] Preparation of (76-3): To a solution of 76-2 (770 mg, 1.68
mmol) in dioxane (10 mL) was added Me.sub.6Sn.sub.2 (1.1 g, 3.36
mmol) and (PPh.sub.3).sub.2PdCl.sub.2 (100 mg) under N.sub.2
atmosphere. The mixture was heated at 80.degree. C. for 4 h. The
mixture was concentrated, and the residue was purified on a silica
gel column to give an intermediate (400 mg, 43.96%). To a solution
of the intermediate (330 mg, 0.61 mmol) in anhydrous MeCN (3 mL)
was added Selectflour.RTM. (462 mg, 1.34 mmol) at R.T. The mixture
was stirred at R.T. for 2 days. The mixture was concentrated, and
the residue was purified on a silica gel column (PE:EA=4:1) to give
76-3 (100 mg, 41.5%).
[0573] Preparation of (76a): To a solution of 76-3 (100 mg, 0.25
mmol) in MeCN (2 mL) was added DMAP (62 mg, 0.51 mmol), TEA (51 mg,
0.51 mmol) and TPSCl (153 mg, 0.51 mmol). The mixture was stirred
at R.T. for 0.5 h. NH.sub.3.H.sub.2O (0.75 mL) was added. The
mixture was stirred at R.T. for 0.5 h. The mixture was extracted
with EtOAc and washed with 1N HCl and brine. The organic layer was
dried and concentrated. The residue was purified on a silica gel
column (PE:EA=1:1) to give an intermediate (60 mg, 60.1%). The
intermediate (50 mg, 0.13 mmol) in NH3/MeOH (5 mL) was stirred at
R.T. for 3 h. The mixture was concentrated, and the residue was
purified on a silica gel column (MeOH:DCM=1:10) to give 76a (30 mg,
76.2%). .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.25 (d, J=6.8
Hz, 1H), 6.09 (d, J=16.0 Hz, 1H), 5.00 (dt, J=4.0 Hz, J=53.2 Hz,
1H), 4.48-4.54 (m, 1H), 3.73-3.95 (m, 4H). ESI-TOF-MS: m/z 312.1
[M+H].sup.+.
Example 74
Preparation of Compound (77a)
##STR00265##
[0575] 77-1 (680 mg, 0.8 mmol) and triphenylphosphine (312 mg, 1.2
mmol) were dissolved in the mixture of 5 mL of dioxine and 0.25 mL
of dry ethanol. A solution of diisopropyl azadicarboxylate (40% w
solution in toluene, 1.28 mmol) in 3 mL of dioxane was added, and
the mixture was stirred at R.T. for 2 h. The mixture was evaporated
to dryness. The residue was dissolved in 10 mL of THF, cooled down
to 4.degree. C. and 2 equivalents of TBAF in THF were added. The
mixture was warmed up to R.T. and the solvent was evaporated. The
resulting nucleoside was treated with 80% HCOOH at R.T. for 3 h,
and then the acid was evaporated. Isolated by isocratic silica gel
chromatography using mixture of DCM (950 mL), MeOH (50 mL), and
NH.sub.4OH (2.5 mL) for elution gave 77a (80 mg, 30%). H.sup.1-NMR
(DMSO-D.sub.6) .delta.: 8.06 (s, 1H), 6.41 (s, 2H), 6.11-6.06 (dd,
1H), 5.98-5.89 (dd, 1H), 5.65-5.64 (d, 1H), 5.34-5.26 (m, 2H),
5.18-5.11 (m, 1H), 4.58-4.50 (dt, 1H), 4.42-4.36 (q, 2H), 3.50-3.28
(m, 2H), 1.30 (t, 3H). MS: 384 (M-1+HCOOH).
Example 75
Preparation of Compound (78a)
##STR00266##
[0577] Preparation of (78-2): To a solution of 78-1 (10.0 g, 37.17
mmol) in anhydrous pyridine (100 mL) was added imidazole (9.54 g,
140.4 mmol) and TBSCl (21.1 g, 140.4 mmol) at 25.degree. C. The
solution was stirred at 25.degree. C. for 15 h. The solution was
concentrated to dryness under reduced pressure. The residue was
dissolved in EtOAc (200 mL) and washed with water and brine. The
organic layer was separated, dried over anhydrous Na.sub.2SO.sub.4
and filtered. The filtrate was concentrated in vacuo to give a
residue. The residue was purified by a silica gel column
(PE/EA=10:1 to 2:1) to give an intermediate (11.8 g, 64%). To an
ice-cold solution of the intermediate (11.8 g, 23.7 mmol) in
CH.sub.2Cl.sub.2 (150 mL) was added a solution of p-toluenesulfonic
acid monohydrate (8.2 g, 47.5 mmol) in small portion under N.sub.2.
The mixture was stirred at 25.degree. C. for 30 min, and then
washed with saturated aq. NaHCO.sub.3. The organic layer was
separated, dried over anhydrous Na.sub.2SO.sub.4 and filtered. The
filtrate was concentrated in vacuum to give a residue, which was
purified by silica gel (PE/EA=10:1 to 1:1) to give 78-2 (6.7 g,
74%) as a solid.
[0578] Preparation of (78-3): To a solution of 78-2 (6.7 g, 17.5
mmol) in anhydrous pyridine (50 mL) was added TMSCl (2.8 g, 26.2
mmol) in small portions at 0.degree. C. under N.sub.2. The reaction
mixture was stirred at 25.degree. C. overnight. AgNO.sub.3 (77.8 g,
510 mmol) and MMTrC (156.8 g, 510 mmol) in anhydrous pyridine (50
mL) was added in small portions under N.sub.2. The reaction mixture
was stirred at 25.degree. C. overnight. Ammonia (30 mL) was added,
and the reaction mixture was stirred for 30 min. The mixture was
filtered through a Buchner funnel, and the filtrate was washed with
saturated NaHCO.sub.3 solution and brine. The organic layer was
separated, dried over anhydrous Na.sub.2SO.sub.4, filtered and
concentrated. Chromatography on silica gel (PE:EA=10:1 to 2:1) gave
an amine protected derivative (6.1 g, 53%). To a solution of
pyridine (142 mg, 1.8 mmol) in anhydrous DMSO (2 mL) at 0.degree.
C. was added TFA (1.3 mg, 0.9 mmol) dropwise. The mixture was
stirred at 25.degree. C. until a clear solution formed. The
solution was then added into a solution of the amine protected
derivative (1.0 g, 1.5 mmol) and DCC (0.95 g, 4.6 mmol) in
anhydrous DMSO at 0.degree. C. dropwise. Stirring was continued at
25.degree. C. for 10 h. Water (10 mL) was added, and the mixture
was stirred at 25.degree. C. for 1 h. The precipitate was removed
by filtration, and the filtrate was extracted with EtOAc (20 mL).
The organic layer was washed with brine (20 mL) and then dried over
Na.sub.2SO.sub.4. The solvent was removed, and the residue was
purified on a silica gel column (EA:PE=10:1 to 2:1) to give the
aldehyde derivative (850 mg, 85%). To a solution of the aldehyde
derivative (2.6 g, 4.0 mmol) in 1,4-dioxane (30 mL) was added 37%
CH.sub.2O (1.3 g, 16.0 mmol) and 2N NaOH aqueous solution (3.0 mL,
6.0 mmol). The mixture was stirred at 25.degree. C. for 2 h and
then neutralized with AcOH to pH=7. To the reaction were added EtOH
(10 mL) and NaBH.sub.4 (912 mg, 24.0 mmol). The reaction was
stirred for 30 mins, and then quenched with saturated aqueous
NH.sub.4Cl. The mixture was extracted with EA, and the organic
layer was dried over Na.sub.2SO.sub.4. Purification by silica gel
column chromatography (EA:PE=10:1 to 2:1) gave 78-3 (1.1 g, 40%) as
a yellow solid.
[0579] Preparation of (78-4): A stirred solution of 78-3 (685 mg,
1.0 mmol) in anhydrous CH.sub.3CN (5 mL) and anhydrous pyridine (5
mL) was cooled to 0.degree. C. BzCl (126 mg, 0.9 mmol) was added,
and the reaction mixture was stirred at 25.degree. C. After 1.5 h,
water (5 mL) was added. The resulting mixture was extracted with
DCM (2.times.30 mL). The combined extracts were washed with a
saturated aqueous solution of NaHCO.sub.3 (20 mL), dried over
MgSO.sub.4, and evaporated under reduced pressure. The residue was
purified by silica gel column chromatography (DCM:MeOH=200:1 to
50:1) to give the Bz-protected derivative (679 mg, 86%). To a
stirred solution of Bz-protected derivative (432 mg, 0.55 mmol) in
anhydrous DMF (5 mL) was added imidazole (258 mg, 3.85 mmol) and
TBSCl (240.0 mg, 1.65 mmol). The mixture was stirred for 15 h.
Water (10 mL) was added, and the mixture was extracted with EA. The
combined extracts were washed with aqueous solution of NaHCO.sub.3
(60 mL) and brine (60 mL), dried over MgSO.sub.4, and evaporated
under reduced pressure to give the two-TBS protected derivative
(680 mg, 137%). The two-TBS protected derivative (680 mg, 0.75
mmol) was dissolved in anhydrous CH.sub.3OH (5 mL), and NaOCH.sub.3
(162 mg, 3.0 mmol) was added. The reaction mixture was stirred at
35.degree. C. for 2 h. The reaction was quenched with 80% AcOH (3
mL) and extracted with DCM (2.times.50 mL). The combined extracts
were washed with aqueous solution of NaHCO.sub.3 (20 mL), dried
over MgSO.sub.4, and evaporated under reduced pressure. The residue
was purified by silica gel column chromatography (EA:PE=20:1 to
3:1) to give 78-4 (239 mg, 40%) as a white foam.
[0580] Preparation of (78-5): 78-4 (239 mg, 0.30 mmol) was
co-evaporated with toluene three times to remove H.sub.2O. To a
solution of 78-4 in DCM (5 mL) was added DMAP (182 mg, 1.50 mmol)
and TfCl (69 mg, 0.45 mmol) at 0.degree. C. under N.sub.2. The
mixture was stirred 0.degree. C. for 40 mins. Completion of the
reaction was determined by LCMS. The mixture was concentrated to
give the crude Tf-derivative (353 mg). To a solution of the
Tf-derivative in DMF (5 mL) was added LiCl (31 mg, 0.76 mmol) at
0.degree. C. under N.sub.2. The mixture was stirred at 25.degree.
C. for 40 mins. The mixture was washed with NaHCO.sub.3 and
extracted with EA. The combined organic layer was dried over
Na.sub.2SO.sub.4 and concentrated to give crude 78-5 (268 mg) as a
light yellow oil.
[0581] Preparation of (78a): To a solution of 78-5 (268 mg, 0.328
mmol) in MeOH (5 mL) was added NH.sub.4F (37 mg, 0.984 mmol) at
25.degree. C. for 4 h. The solution was filtered and evaporated to
dryness. The residue was dissolved in HCOOH (20 mL) and H.sub.2O (4
mL) at 25.degree. C. The mixture was stirred at 25.degree. C. for 1
h and concentrated. The mixture was dissolved in MeCN and purified
by prep-HPLC to give 78a (32 mg) as a white solid. .sup.1H NMR
(MeOD, 400 MHz) .delta. 8.33 (s, 1H), 8.20 (s, 1H), 6.32 (dd,
J=5.6, 12.4 Hz, 1H), 5.77 (m, 1H), 4.69 (m, 1H), 3.85 (m, 1H).
ESI-MS: m/z 317.9 [M+H].sup.+.
Example 76
Preparation of Compound (79a)
##STR00267##
[0583] Preparation of (79-1): To a solution of 78-4 (1.1 g, 1.33
mmol) in anhydrous DCM (6.6 mL) at 0.degree. C. under nitrogen was
added Dess-Martin periodinane (1.45 g, 3.33 mol). The mixture was
stirred at 25.degree. C. for 4 h. The solvent was removed in
vacuum, and the residue triturated with methyl-t-butyl ether (30
mL). The mixture was filtered through a pad of MgSO.sub.4, and the
organic solvent was stirred with an equal volume of
Na.sub.2S.sub.2O.sub.3 in 30 mL of saturated NaHCO.sub.3 until the
organic layer became clear (approx. 10 min). The organic layer was
separated, washed with brine, and dried over MgSO.sub.4. Prior to
removing the solvent in vacuum, the residue was purified on a
silica gel column (PE:EA=7:1) to give 79-1 (750 mg, 75%) as a white
solid.
[0584] Preparation of (79-2): To a stirred solution of
methyl-triphenyl-phosphonium bromide (1.74 g, 4.89 mmol) in
anhydrous THF (8 mL) was added n-BuLi (1.91 mL, 4.89 mmol, 2.5 M in
THF) at -78.degree. C. dropwise. The mixture was stirred at
0.degree. C. for 1 h. 79-1 (750 mg, 0.81 mmol) was added, and the
mixture stirred at 25.degree. C. overnight. The reaction was
quenched with saturated NH.sub.4Cl (30 mL), and extracted with
EtOAc (2.times.30 mL). The combined organic phase was washed with
brine, dried with MgSO.sub.4, filtered and evaporated to dryness to
give a light white solid. The solid was purified by column
chromatography (PE:EA=5:1) to give 79-2 (440 mg, 60%).
[0585] Preparation of (79-3): To a solution of 79-2 (440 mg, 0.48
mmol) in MeOH (8 mL) was added Pd/C (500 mg, 10%) at R.T. under
hydrogen atmosphere. The mixture was stirred at R.T. for 1.5 h. The
mixture was filtered, and the filtrate was concentrated to dryness.
Crude 79-3 (365 mg, 83%) was used for the next step without further
purification.
[0586] Preparation of (79a): 79-3 (365 mg, 0.40 mmol) in MeOH (50
mL) was added NH.sub.4F (5.6 g, 0.15 mmol), and the solution was
heated to refluxed overnight. Completion of the reaction was
determined by LCMS. The mixture was filtered, and the filtrate was
concentrated to dryness. The residue was purified on a silica gel
column (PE:EA=3:1) to give the amine protected derivative (173 mg,
77%) as a white solid. The amine protected derivative (100 mg, 0.18
mmol) in formic acid (4.4 mL) was stirred at 25.degree. C.
overnight. The solution was concentration to dryness, and the
residue was purified on a silica gel column (PE:EA=1:3) to give 79a
(40 mg, 90%) as a white solid. .sup.1H NMR (400 MHz, CD.sub.3OD)
8.25 (s, 1H), 8.09 (s, 1H), 6.14 (dd, J=6.0, 12.8 Hz, 1H), 5.58 (m,
1H), 4.45-4.48 (m, 1H), 3.60 (q, 2H), 1.66-1.74 (m, 2H), 0.88 (t,
3H); ESI-MS: m/z 297.9 [M+H].sup.+.
Example 77
Preparation of Compound (80a)
##STR00268##
[0588] Preparation of (80-1): To a solution of 78-3 (4.4 g, 6.4
mmol) in anhydrous pyridine (5 mL) and DCM (25 mL). A solution of
DMTrCl (2.37 g, 7.04 mmol) in DCM (5 mL) was added dropwise at
0.degree. C. under N.sub.2. After 2 h, the reaction was quenched
with CH.sub.3OH and concentrated to dryness. The residue was
purified on a column of silica gel (PE:EA=100:1 to 2:1) to obtain
the DMTr protected derivative (4.3 g, 68%). The DMTr protected
derivative (2.2 g, 2.5 mmol) in 1M TBAF (2.5 mL) of THF (2.5 mL)
solution was stirred at 25.degree. C. for 3 h. The solvent was
removed in vacuum, and the residue was purified by column
chromatography (PE/EA=50:1 to 1:2) to give the diol derivative
(1.86 g, 96%). To a solution of the diol derivative (1.3 g, 1.5
mmol) in anhydrous THF (5 mL) was added NaH (132 mg, 3.3 mmol) at
0.degree. C. The mixture was stirred for 1 h, and TBI (276 mg, 0.75
mmol), and BnBr (558 mg, 3.3 mmol) was added. The mixture was
stirred for 10 h at 25.degree. C. The reaction was quenched with
water, and the solvent was evaporated. The mixture was extracted
with EA and brine. The organic layer was dried over
Na.sub.2SO.sub.4, and evaporated to afford the crude product. The
product was purified by silica gel (PE/EA=100:1 to 3:1) to afford
80-1 (1.4 g, 90%) as a white foam.
[0589] Preparation of (80-2): To a solution of 80-1 (1.3 g, 1.23
mmol) in anhydrous DCM (17 mL) was added Cl.sub.2CHCOOH (1.57 g,
12.3 mmol) at -78.degree. C. The mixture was stirred at
-20-10.degree. C. for 40 mins. The reaction was quenched with
saturated NaHCO.sub.3, and diluted with DCM (50 mL). The mixture
was washed with brine, and the organic solution was dried over
Na.sub.2SO.sub.4 and concentrated in vacuum. The residue was
purified on a silica gel column (PE/EA=100:1 to 1:1) to give 80-2
(652 mg, 70%) as a white foam.
[0590] Preparation of (80-3): To a solution of 80-2 (630 mg, 0.84
mmol) in anhydrous DCM (5 mL) was added DAST (1.35 g, 8.4 mmol) at
-78.degree. C. The mixture was gradually warmed to 0.degree. C. The
reaction was quenched with saturated NaHCO.sub.3. The mixture was
diluted with DCM (50 mL) and washed with brine. The organic
solution was dried over Na.sub.2SO.sub.4 and concentrated in
vacuum. The residue was purified on a silica gel column
(PE/EA=100:1 to 2:1) to give 80-3 as a white solid (302 mg,
48%).
[0591] Preparation of (80a): A mixture of 80-3 (210 mg, 0.28 mmol)
and Pd(OH).sub.2 (200 mg) in methanol (3 mL) was stirred at
0.degree. C. at 40 psi H.sub.2 for 20 h. Pd(OH).sub.2 was filtered
off, and the filtrate was concentrated to dryness. The residue was
purified by column (DCM/MeOH=10:1) to give 80a (12 mg). .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 8.33 (s, 1H), 8.20 (s, 1H), 6.33 (dd,
J=6.0, 13.2 Hz, 1H), 5.79 (t, J=5.6 Hz, 1H), 5.66 (t, J=5.2 Hz,
1H), 4.52-4.80 (m, 3H), 3.80-3.82 (m, 2H). ESI-MS: m/z 302.0
[M+H].sup.+.
Example 78
Preparation of Compound (81a)
##STR00269##
[0593] Preparation of (81-2): To a solution of 81-1 (20.0 g, 70.2
mmol) in anhydrous pyridine (200 mL) was added imidazole (19.1 g,
280 mmol) and TBSCl (42.1 g, 281 mmol) at 25.degree. C. The
solution was stirred at 25.degree. C. for 15 h, and then
concentrated to dryness under reduced pressure. The residue was
dissolved in EtOAc and then filtered. The filtrate was concentrated
to dryness to give the TBS protected derivative (36.4 g, 99%). The
TBS protected derivative (36.5 g, 71.1 mmol) was dissolved in THF
(150 mL). H.sub.2O (100 mL), and then AcOH (300 mL) were added. The
solution was stirred at 80.degree. C. for 13 h. The reaction was
cooled to R.T., and then concentrated to dryness under reduced
pressure to give 81-2 (31.2 g, 61%) as a white solid.
[0594] Preparation of (81-3): To a solution of 81-2 (31.2 g, 78.2
mmol) in anhydrous pyridine (300 mL) was added Ac.sub.2O (11.9 g,
117.3 mmol). The mixture was stirred at 25.degree. C. for 18 h.
MMTrC (72.3 g, e234.6 mmol) and AgNO.sub.3 (39.9 g, 234.6 mmol)
were added, and the solution was stirred at 25.degree. C. for 15 h.
H.sub.2O was added to quench the reaction and the solution was
concentrated to dryness under reduced pressure. The residue was
dissolved in EtOAc and washed with water. The organic layer was
dried over Na.sub.2SO.sub.4 and filtered. The filtrate was
concentrated in vacuum to give a residue, which was purified by
silica gel (DCM:MeOH=200:1 to 50:1) to give the MMTr protected
amine derivative (35.2 g, 63%). The MMTr protected amine derivative
(35.2 g, 49.3 mmol) was dissolved in NH3/MeOH (300 mL). The mixture
was stirred at 25.degree. C. for 20 h. The solution was evaporated
to dryness, and purified by a silica gel column (DCM:MeOH=100:1 to
50:1) to give 81-3 as a yellow solid (28.6 g, 87%).
[0595] Preparation of (81-4): To a solution of 81-3 (12.0 g, 17.9
mmol) in anhydrous DCM (200 mL) was added Dess-Martin periodinane
(11.3 g, 26.8 mmol) at 0.degree. C. The mixture was stirred at
0.degree. C. for 2 h, and then at R.T. for 2 h. The mixture was
quenched with a saturated NaHCO.sub.3 and Na.sub.2S.sub.2O.sub.3
solution. The organic layer was washed with brine (2.times.) and
dried over anhydrous Na.sub.2SO.sub.4. The solvent was evaporated
to give the aldehyde (12.6 g), which was used directly in the next
step. To a solution of the aldehyde (12.6 g, 18.0 mmol) in
1,4-dioxane (120 mL) was added 37% HCHO (11.6 g, 144 mmol) and 2N
NaOH aqueous solution (13.5 mL, 27 mmol). The mixture was stirred
at 25.degree. C. overnight. EtOH (60 mL) and NaBH.sub.4 (10.9 g,
288 mmol) were added, and the reaction was stirred for 30 mins. The
mixture was quenched with saturated aqueous NH.sub.4Cl, and then
extracted with EA. The organic layer was dried over
Na.sub.2SO.sub.4, and purified by silica gel column chromatography
(DCM:MeOH=200:1 to 50:1) to give 81-4 (7.5 g, 59%) as a yellow
solid.
[0596] Preparation of (81-5): To a solution of 81-4 (3.8 g, 5.4
mmol) in DCM (40 mL) was added pyridine (10 mL) and DMTrCl (1.8 g,
5.4 mmol) at 0.degree. C. The solution was stirred at 25.degree. C.
for 1 h. MeOH (15 mL) was added, and the solution was concentrated.
The residue was purified by silica gel column chromatography
(DCM:MeOH=200:1 to 50:1) to give the MMTr protected derivative (3.6
g, 66%) as a yellow solid. To a solution of the MMTr protected
derivative (3.6 g, 3.6 mmol) in anhydrous pyridine (30 mL) was
added TBDPSCl (2.96 g, 10.8 mmol) and AgNO.sub.3 (1.84 g, 10.8
mmol). The mixture was stirred at 25.degree. C. for 15 h. The
mixture was filtered and concentrated. The mixture was dissolved in
EtOAc and washed with brine. The organic layer was dried over
Na.sub.2SO.sub.4, and then purified by silica gel column
chromatography (DCM:MeOH=200:1 to 50:1) to give the TBDPS protected
derivative (3.8 g, 85.1%) as a solid. To a solution of the TBDPS
protected derivative (3.6 g, 2.9 mmol) in anhydrous DCM (50 mL) was
added Cl.sub.2CHCOOH (1.8 mL) in anhydrous DCM (18 mL). The mixture
was stirred at -78.degree. C. for 1 h. Cl.sub.2CHCOOH (3.6 mL) was
added at -78.degree. C. The mixture was stirred at -10.degree. C.
for 30 mins. The mixture was quenched with saturated aqueous
NaHCO.sub.3 and extracted with DCM. The organic layer was dried
over Na.sub.2SO.sub.4, and then purified by silica gel column
chromatography (DCM:MeOH=200:1 to 50:1) to give 81-5 (2.2 g,
80%).
[0597] Preparation of (81-6): To an ice cooled solution of 81-5
(800 mg, 0.85 mmol) in anhydrous DCM (20 mL) was added pyridine
(336 mg, 4.25 mmol) and Tf.sub.2O (360 mg, 1.28 mmol) dropwise. The
reaction mixture was stirred at 0.degree. C. for 15 mins. The
reaction was quenched by ice water and stirred for 30 mins. The
mixture was extracted with EtOAc, washed with brine (50 mL) and
dried over MgSO.sub.4. The solvent was evaporated to give the crude
bis(triflate) derivative. To the bis(triflate) derivative (790 mg,
0.73 mmol) in anhydrous DMF (35 mL) was added LiCl (302 mg, 7.19
mmol). The mixture was heated to 40.degree. C. and stirred
overnight. Completion of the reaction was determined by LCMS. The
solution was washed with brine and extracted with EtOAc. The
combined organic layers were dried over MgSO.sub.4, and the residue
was purified on a silica gel column (DCM/MeOH=100:1) to give 81-6
(430 mg, 61%).
[0598] Preparation of (81a): To 81-6 (470 mg, 0.49 mmol) in MeOH
(85 mL) was added NH.sub.4F (8.1 g, 5.92 mmol), and the solution
was heated to reflux overnight. The mixture was filtered, and the
filtrate was concentrated to dryness. The residue was purified on a
silica gel column (DCM/MeOH=20:1) to give the diol (250 mg, 84%) as
a white solid. The diol (130 mg, 0.21 mmol) in formic acid (5 mL)
was stirred at 25.degree. C. overnight. The solution was
concentration to dryness, and the residue in MeOH (30 mL) was
stirred at 70.degree. C. overnight. Completion of the reaction was
determined by LCMS and HPLC. The solvent was removed, and the crude
product was washed with EtOAc to give 81a (58 mg, 81%) as a white
solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 10.73 (br, 1H),
7.98 (s, 1H), 6.58 (br, 2H), 6.08 (q, J=4.8, 9.2 Hz, 2H), 5.64 (dt,
J=5.6, 52.8 Hz, 1H), 5.40 (m, 1H), 4.52 (m, 1H), 3.80-3.82 (m, 2H),
3.64 (q, 2H). ESI-MS: m/z 333.8 [M+H].sup.+, 666.6 [2M+H]+
Example 79
Preparation of Compound (82a)
##STR00270##
[0600] Preparation of (82-1): To a solution of 81-4 (310 mg, 0.33
mmol) in anhydrous DCM (10 mL) was added pyridine (130 mg, 1.65
mmol) and Tf.sub.2O (139 mg, 0.49 mmol) diluted by DCM dropwise at
0.degree. C. The mixture was stirred at 0.degree. C. for 15 mins.
The reaction was quenched with ice cold water. The organic layer
was separated and washed with brine. The organic layer was dried
over Na.sub.2SO.sub.4 and evaporated to give to give the triflate
derivative (420 mg crude), which was used directly in the next
step. To a solution of the triflate derivative (420 mg crude) in
anhydrous pentan-2-one was added NaI (396 mg, 2.64 mmol). The
mixture was stirred at 40.degree. C. for 3 h, and then dissolved
with EtOAc. The organic layer were washed with
Na.sub.2S.sub.2O.sub.3 twice and washed with brine. The organic
layer was dried over Na.sub.2SO.sub.4 and evaporated to give a
residue. The residue was purified by a column (DCM:MeOH=300:1 to
100:1) to give 82-1 (195 mg, 56% for two steps).
[0601] Preparation of (82-2): To a solution of 82-1 (650 mg, 0.62
mmol) in MeOH (10 mL) was added NH.sub.4F (45.8 g, 12.4 mmol). The
mixture was refluxed overnight. The mixture was filtered and
evaporated to dryness. The residue was purified on a silica gel
column (DCM/MeOH=200:1 to 20:1) to give 82-2 (250 mg, 58%).
[0602] Preparation of (82-3): To a stirred solution of 82-2 (300
mg, 0.43 mmol), Et.sub.3N (217 mg, 2.15 mmol) in anhydrous MeOH (10
mL) was added 10% Pd/C (50 mg). The mixture was stirred in a
hydrogenation apparatus (30 psi hydrogen) at R.T. overnight. The
catalyst was filtrated off, and the filtrate was evaporated to give
a residue. The residue was purified on a silica gel column
(DCM/MeOH=200:1 to 20:1) to afford 82-3 as a white solid (180 mg,
73%).
[0603] Preparation of (82a): 82-3 (110 mg, 0.19 mmol) was dissolved
in HCOOH (18 g) and H.sub.2O (6 g) at 25.degree. C., and stirred
for 1 h. The solution was evaporated to dryness, dissolved in MeOH
(30 mL). The mixture was stirred at 60.degree. C. for 12 h. The
solution was evaporated to dryness, and dissolved in EtOAc (50 mL).
The mixture was stirred at 60.degree. C. for 1 h. The mixture was
filtered and washed with EtOAc to give 82a as a white solid (45.3
mg, 80%). .sup.1H NMR (400 MHz, MeOD) .delta. 8.00 (s, 1H),
6.11-6.15 (m, 1H), 5.35-5.50 (m, 1H), 4.53-4.59 (m, 1H), 3.54-3.64
(m, 2H), 1.26 (s, 3H). ESI-MS: m/z 299.76 [M+1]+, 598.66
[2M+1]+.
Example 80
Preparation of Compound (83a)
##STR00271##
[0605] Preparation of (83-1): 81-1 (5.7 g. 20 mmol) was
co-evaporated with pyridine three times, and then dissolved in
pyridine (20 mL). The mixture was cooled to 0.degree. C. and
Ac.sub.2O (5.8 mL, 60 mmol) was added dropwise. The mixture was
stirred at 25.degree. C. for 10 h, and then cooled to 0.degree. C.
AgNO.sub.3 (8.5 g, 50 mmol), and then MMTrCl (15.5 g, 50 mmol) were
added in portions. The mixture was stirred at 25.degree. C. for 10
h. The reaction was quenched with saturated NaHCO.sub.3 and
extracted with EA. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified by
silica gel column chromatography (DCM/MeOH=100:1 to 50:1) to afford
the Ac protected derivative (12.1 g, 93%) as a light yellow solid.
The Ac protected derivative (12.1 g) was dissolved in methanolic
NH.sub.3 (saturated). The mixture was stirred at 25.degree. C. for
14 h. The solvent was removed, and the residue was purified on a
silica gel column (DCM/MeOH=80:1 to 30:1) to give 83-1 (9.2 g,
87%).
[0606] Preparation of (83-2): To a stirred solution of 83-1 (9.2 g,
16.5 mmol) in dry THF (300 mL) was added imidazole (9.0 g, 132
mmol) and PPh.sub.3 (34.8 g, 132 mmol). A solution of I.sub.2 (26.0
g, 103 mmol) in THF (100 mL) was added dropwise under N.sub.2 at
0.degree. C. The mixture was stirred at 25.degree. C. for 18 h and
then quenched with a Na.sub.2S.sub.2O.sub.3 solution. The mixture
was extracted with EtOAc. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. The residue was purified on a
silica gel column (DCM/MeOH=80:1 to 30:1) to give the iodide
derivative (10.3 g, 93%) as a light yellow solid. To a stirred
solution of the iodide derivative (10.2 g, 15.3 mmol) in dry THF
(300 mL) was added DBU (4.7 g, 30.1 mmol). The mixture was stirred
at 60.degree. C. for 8 h. The solution was diluted with a
NaHCO.sub.3 solution and extracted with EtOAc. The organic layer
was dried over Na.sub.2SO.sub.4 and concentrated. The residue was
purified on a silica gel column (PE/EtOAc=3:1 to 1:3) to afford
83-2 (6.2 g, yield 76%).
[0607] Preparation of (83-3): To a stirred solution of 83-2 (5.42
g, 10 mmol) in anhydrous CH.sub.3H (100 mL) was added PbCO.sub.3
(13.7 g, 53.1 mmol). A solution of I.sub.2 (12.3 g, 48.9 mmol) in
CH.sub.3OH (300 mL) was added dropwise at 0.degree. C. The mixture
was stirred at 25.degree. C. for 10 h. The solution was quenched
with a Na.sub.2S.sub.2O.sub.3 solution and extracted with DCM. The
organic layer was washed with a NaHCO.sub.3 solution, dried over
Na.sub.2SO.sub.4 and concentrated to give a residue. The residue
was purified by HPLC (0.1% HCOOH in water and MeCN) to give the
desired methoxyl derivative (2.4 g, 34%). To a stirred solution of
the methoxyl derivative (2.4 g, 3.4 mmol) in dry pyridine (20 mL)
was added BzCl (723 mg, 5.2 mmol) dropwise at 0.degree. C. The
mixture was stirred at 0.degree. C. for 1 h. The solution was
quenched with a NaHCO.sub.3 solution and extracted with EtOAc. The
organic layer was dried over Na.sub.2SO.sub.4 and concentrated.
Purified by a silica gel column (PE/EtOAc=5:1 to 1:1) afforded 83-3
(2.1 g, 77%) as a white solid.
[0608] Preparation of (83a): 83-3 (2.0 g, 2.5 mmol), BzONa (3.6 g,
25 mmol) and 15-crown-5 (5.5 g, 25 mmol) were suspended in DMF (50
mL). The mixture was stirred at 110-125.degree. C. for 5 days. The
precipitate was removed by filtration, and the filtrate was diluted
with EA. The solution was washed with brine and dried over
Na.sub.2SO.sub.4. The solvent was removed, and the residue was
purified on a silica gel column (PE/EA=10/1 to 2/1) to afford the
crude Bz protected derivative (1.6 g, 80%). The Bz protected
derivative (1.6 g, 2.0 mmol) was dissolved in methanolic ammonia
(100 mL), and the mixture was stirred at 25.degree. C. for 20 h.
The solvent was removed, and the residue was purified by a silica
gel column (DCM/MeOH=100:1 to 20:1) to the diol derivative as a
white solid (410 mg, 35%). The diol derivative (200 mg, 0.34 mmol)
was dissolved in HCOOH (24 g) and H.sub.2O (6 g) at 25.degree. C.,
and the mixture was stirred at 25.degree. C. for 1 h. The solution
was evaporated to dryness, and dissolved in MeOH (30 mL). The
mixture was stirred at 60.degree. C. for 12 h. The solution was
evaporated to dryness and dissolved in EtOAc (50 mL). The mixture
was stirred at 60.degree. C. for 1 h. The mixture was then filtered
and washed with EtOAc to give 83a as a white solid (46.1 mg, 43%).
H NMR (CD.sub.3OD, 400 MHz) .delta. 7.92 (s, 1H), 6.22 (dd, J=1.6,
18.8 Hz, 1H), 5.17-5.32 (m, 1H), 4.89-4.91 (m, 1H), 3.77 (m, 2H),
3.44 (s, 3H). ESI-MS: m/z 316.1 [M+H].sup.+.
Example 81
Preparation of Compound (84a)
##STR00272## ##STR00273##
[0610] Preparation of (84-2): To a stirred solution of 84-1 (100.0
g, 265.9 mmol) in dry THF (1000 mL) was added Li(O-t-Bu).sub.3AlH
(318.9 mL, 318.9 mmol) at -78.degree. C. under N.sub.2. The mixture
was stirred at -78.degree. C. for 1 h and then at R.T for 1 h. The
reaction mixture was cooled to -50.degree. C. and quenched with ice
and a saturated NH.sub.4Cl solution. The mixture was extracted with
EtOAc. The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated to afford the 1'-OH derivative (100.5 g) as a white
solid. To a stirred solution of the 1'-OH derivative (100.5 g,
265.9 mmol) in dry DCM (600 mL), NEt.sub.3 (110 mL) and MsCl (45.5
g, 298.0 mmol) were added dropwise at 0.degree. C. The mixture was
stirred at R.T. for 2 h. The mixture was quenched with ice water at
0.degree. C. and extracted with DCM. The organic layer was dried
over Na.sub.2SO.sub.4, concentrated and purified on a silica gel
column (PE:EA=50:1 to 5:1) to afford 84-2 (113.4 g, yield: 93.9%)
as a white solid.
[0611] Preparation of (84-3): To a suspension of compound
6-chloro-9H-purin-2-amine (70.1 g, 414.7 mmol), HMDS (480 mL) and
(NH.sub.4).sub.2SO.sub.4 (0.8 g) was added dry DCE (400 mL). The
mixture was refluxed under N.sub.2 for 18 h and then cooled to R.T.
To the silylated 2-amino-6-chloropurine solution was added 84-2
(78.0 g, 171.1 mmol) and TMSOTf (60 mL, 331.9 mmol). The mixture
was refluxed overnight, concentrated and neutralized with a
NaHCO.sub.3 solution. The resulting precipitate was filtered, and
the filtrate was extracted with EtOAc. The organic layer was dried
over Na.sub.2SO.sub.4 and concentrated. Chromatography on a silica
gel column (PE:EA=5:1 to 2:1) gave 84-3 (10.8 g, yield: 11.9%) as a
light yellow solid.
[0612] Preparation of (84-4): To a suspension of 84-3 (30.0 g, 56.6
mmol) in DCM (300 mL) were added MMTrCl (34.9 g, 113.2 mmol) and
AgNO.sub.3 (19.3 g, 113.2 mmol). The reaction mixture was cooled to
0.degree. C., and collidine (18.0 g, 150 mmol) was added. The
resulting suspension was stirred at R.T. for 12 h. The suspension
was filtered. The filtrate was extracted with DCM and washed with a
NaHCO.sub.3 solution. The organic layer was dried over
Na.sub.2SO.sub.4 and concentrated. Purification by a silica gel
column (PE:EA=20:1 to 3:1) to give 84-4 (35.0 g, yield: 77.9%) as a
light yellow solid. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
7.94-7.96 (m, 4H), 7.05-7.58 (m, 18H), 6.62-6.67 (m, 2H), 6.55 (dd,
J=6.0 Hz, J=9.6 Hz, 1H), 5.60-5.66 (m, 1H), 4.69-4.76 (m, 2H),
4.55-4.58 (m, 1H), 3.64 (s, 1H). ESI-MS: m/z 802 [M+H].sup.+.
[0613] Preparation of (84-5 To a stirred solution of 84-4 (35.0 g,
43.6 mmol) in dry MeOH (400 mL) was added NaOMe (23.5 g, 436 mmol)
and 2-mercapto-ethanol (30.6 g, 392.4 mmol). The mixture was
refluxed overnight. The pH was adjusted to 9-10 with CO.sub.2. The
precipitate was filtered, and the filtrate was concentrated.
Purification on a silica gel column (PE:EA=10:1 to 1:1) gave pure
84-5 (24.0 g, yield 95.7%) as a light yellow solid.
[0614] Preparation of (84-6): To a solution of 84-5 (24.0 g, 41.7
mmol) in pyridine (250 mL) was added DMTrCl (28.2 g, 83.5 mmol) at
0.degree. C. The solution was stirred at R.T. for 15 h. MeOH (50
mL) was added, and the mixture was concentrated to dryness under
reduced pressure. The residue was dissolved in EtOAc and washed
with water. The organic layer was dried over Na.sub.2SO.sub.4,
filtered, concentrated and purified by a silica gel column
(DCM:MeOH=200:1 to 50:1) to give a first intermediate (27.6 g) as a
yellow solid. To a solution of the first intermediate (27.6 g, 31.5
mmol) in DCM (200 mL) was added imidazole (4.3 g, 63 mmol) and
TBSCl (9.5 g, 63 mmol). The mixture was stirred at R.T. for 12 h.
The solution was washed with NaHCO.sub.3 and brine. The organic
layer was dried over Na.sub.2SO.sub.4, filtered, concentrated and
purified by a silica gel column (DCM:MeOH=200:1 to 100:1) to give a
second intermediate (30.2 g) as a yellow solid. To a solution of
the second intermediate (30.2 g, 30.4 mmol) in anhydrous DCM (50
mL) was added Cl.sub.2CHCOOH (20 ml) in anhydrous DCM (500 mL). The
mixture was stirred at -78.degree. C. for 1 h. Cl.sub.2CHCOOH (30
mL) was added at -78.degree. C. The mixture was stirred at
-20.degree. C. for 2 h. The mixture was quenched with saturated
aqueous NaHCO.sub.3 and extracted with DCM. The organic layer was
dried over Na.sub.2SO.sub.4, and then purified by a silica gel
column (DCM:MeOH=200:1 to 30:1) to give 84-6 (18.0 g, 62.5%) as a
white solid. .sup.1H NMR (400 MHz, MeOD) .delta. 8.27 (s, 1H),
7.16-7.38 (m, 12H), 6.79-6.83 (m, 2H), 6.42 (dd, J=4.4 Hz, J=10.0
Hz, 1H), 4.54-4.62 (m, 1H), 3.92 (d, J=8.8 Hz, 2H), 3.74 (s, 3H),
3.70-3.72 (m, 1H), 0.92 (s, 9H), 0.11-0.13 (m, 6H). ESI-LCMS: m/z
690.0 [M+H].sup.+.
[0615] Preparation of (84-7): 84-6 (7.0 g, 10.0 mmol) was added to
a suspension of DMP (10.6 g, 25 mmol) in anhydrous CH.sub.2Cl.sub.2
(100 mL) at 0.degree. C. The mixture was stirred at 25.degree. C.
for 2 h. The solvent was removed in vacuo, and the residue
triturated with diethyl ether (100 mL). The mixture was filtered
through a pad of MgSO.sub.4. The organic solvent was stirred with
an equal volume of Na.sub.2S.sub.2O.sub.3.5H.sub.2O in 100 mL of
saturated NaHCO.sub.3 until the organic layer became clear (10
min). The organic layer was separated, washed with brine, and dried
over MgSO.sub.4. The solvent was removed in vacuo to give a third
intermediate as a red solid (6.5 g, 95%). To a solution of the
third intermediate (6.5 g, 9.5 mmol) in 1,4-dioxane (80 mL) was
added 37% CH.sub.2O (6.0 mL, 60 mmol) and 2N NaOH aqueous solution
(9.5 mL, 19 mmol). The mixture was stirred at 25.degree. C. for 2 h
and then neutralized with AcOH to pH 7. EtOH (30 mL) and NaBH.sub.4
(3.8 g, 100 mmol) were added, and the mixture was stirred for 30
mins. The mixture was quenched with saturated aqueous NH.sub.4Cl,
and then extracted with EA. The organic layer was dried over
Na.sub.2SO.sub.4. Purification by a silica gel column
(DCM:MeOH=200:1 to 30:1) gave 84-7 (4.2 g, 58.3%) as a yellow
solid.
[0616] Preparation of (84-8): To a solution of 84-7 (4.2 g, 5.8
mmol) in DCM (50 mL) was added pyridine (5 mL) and DMTrCl (1.9 g,
5.8 mmol) at -20.degree. C. The solution was stirred at 0.degree.
C. for 2 h. The reaction mixture was treated with MeOH (15 mL), and
then concentrated. The residue was purified by a silica gel column
(DCM:MeOH=200:1 to 50:1) to give the fourth intermediate (1.3 g) as
a yellow solid. To a solution of the fourth intermediate (1.3 g,
1.3 mmol) in anhydrous pyridine (15 mL) was added TBDPSCl (1.1 g,
3.9 mmol) and AgNO.sub.3 (0.68 g, 4.0 mmol). The mixture was
stirred at 25.degree. C. for 15 h. The mixture was filtered,
concentrated, dissolved in EtOAc and washed with brine. The organic
layer was dried over Na.sub.2SO.sub.4. Purification by a silica gel
column (DCM:MeOH=200:1 to 100:1) gave a fifth intermediate (1.4 g)
as a solid. To a solution of the fifth intermediate (1.4 g, 1.1
mmol) in anhydrous DCM (50 mL) was added Cl.sub.2CHCOOH (0.7 ml) in
anhydrous DCM (18 mL). The mixture was stirred at -78.degree. C.
for 1 h. Cl.sub.2CHCOOH (1.5 ml) was added at -78.degree. C., and
the mixture was stirred at -20.degree. C. for 1.5 h. The mixture
was quenched with saturated aqueous NaHCO.sub.3 and extracted with
DCM. The organic layer was dried over Na.sub.2SO.sub.4.
Purification by a silica gel column (DCM:MeOH=200:1 to 50:1) gave
84-8 (650 mg, 11.6%) as a white solid.
[0617] Preparation of (84-9): To a solution of pyridine (521 mg,
6.59 mmol) in anhydrous DMSO (5 mL) was added TFA (636 mg, 5.58
mmol) dropwise at 10.degree. C. under N.sub.2. The mixture was
stirred until a clear solution formed. To this solution (0.8 mL)
was added a mixture of 84-8 (650 mg, 0.68 mmol) and DCC (410 mg,
2.0 mmol) in anhydrous DMSO (5 mL) at R.T. under N.sub.2. The
mixture was stirred at 20.degree. C. overnight. Water (30 mL) was
added. The mixture was diluted with DCM (30 mL) and filtered. The
filtrate was extracted with DCM. The organic layers were washed
with saturated aqueous NaHCO.sub.3, dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The crude product was purified on a silica
gel column (PE:EA=10:1 to 1:1) to give the sixth intermediate (600
mg) as a yellow solid. To a stirred solution of
Methyl-triphenyl-phosphonium bromide (714 mg, 2.0 mmol) in
anhydrous THF (5 mL) was added n-BuLi (0.8 mL, 2.0 mmol, 2.5 M in
THF) at -78.degree. C. dropwise over 1 min. Stirring was continued
at 0.degree. C. for 1 h. The sixth intermediate (600 mg, 0.63 mmol)
was added to the mixture, and the mixture was stirred at 25.degree.
C. for 15 h. The reaction was quenched with saturated NH.sub.4Cl
(20 mL) and extracted with EtOAc. The combined organic phase was
dried with Na.sub.2SO.sub.4, filtered and evaporated to dryness to
give a light yellow oil. The oil was purified by column
chromatography (DCM:MeOH=200:1 to 50:1) to give 84-9 (250 mg,
38.5%) as a yellow solid.
[0618] Preparation of (84-10): 84-9 (250 mg, 0.26 mmol) was
dissolved in THF (5.0 mL). TBAF (131 mg, 0.5 mmol) was added at
20.degree. C., and stirring was continued for 2 h. The solution was
evaporated to dryness. The residue was dissolved in EA (50 mL) and
washed with water (2.times.). The solution was evaporated to
dryness, and purified by a silica gel column (PE:EA=10:1 to 1:2) to
give 84-10 (57.6 mg, 36.9%) as a white solid. .sup.1H NMR (400 MHz,
MeOD) .delta. 8.34 (s, 1H), 7.15-7.38 (m, 12H), 6.79-6.82 (m, 2H),
6.44 (dd, J=2.0 Hz, J=10.0 Hz, 1H), 6.01 (dd, J=11.2 Hz, J=17.6 Hz,
1H), 5.51 (dd, J=1.6 Hz, J=17.2 Hz, 1H), 5.35 (dd, J=1.6 Hz, J=17.2
Hz, 1H), 4.68-4.76 (m, 1H), 3.74 (s, 3H), 3.63 (dd, J=2.0 Hz,
J=12.8 Hz, 1H) 3.52 (dd, J=2.0 Hz, J=12.8 Hz, 1H). ESI-LCMS: m/z
602.0 [M+H].sup.+.
[0619] Preparation of (84a): A solution of 84-10 (27 mg) in 1.5 mL
of 80% formic acid stood at R.T. for 4.5 h and then concentrated to
dryness. The residue was mixed with water and lyophilized. MeOH
(1.5 mL) and TEA (0.1 mL) were added, and the mixture was
concentrated. The precipitate from MeOH and EtOAc was filtered and
washed with EtOAc to give 84 (9.3 mg) as a slightly-amber solid.
.sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.44 (s, 1H), 6.57 (d,
J=10.8 Hz, 1H), 6.05 (dd, J=17.6 Hz, 10.8 Hz, 1H), 5.45 (dd, J=17.6
Hz, J=1.6 Hz, 1H), 5.37 (dd, J=10.8 Hz, 1.6 Hz, 1H), 4.78 (dd,
J=18.4 Hz, 17.2 Hz, 1H), 3.67 (d, J=12.4 Hz, 1H), 3.56 (dd, J=12.4
Hz, 2.0 Hz, 1H); ESI-MS: m/z 328.4 [M-H].sup.-.
Example 82
Preparation of Compound (85a)
##STR00274##
[0621] Preparation of (85-2): A mixture of 85-1 (200 mg; 0.22 mmol)
in pyridine (2.5 mL) and isobutyric anhydride (44 .mu.L; 1.2 equiv)
was stirred R.T. overnight. The mixture was concentrated, and the
residue partitioned between EtOAc (50 mL) and water. The organic
layer was washed with 1N citric acid, water, saturated aqueous
NaHCO.sub.3 and brine. The mixture was dried with Na.sub.2SO.sub.4.
The solvent was evaporated and the residue was purified on a silica
column (10 g column) using hexanes/EtOAc (30 to 100% gradient) to
give 85-2 (0.16 g, 75%).
[0622] Preparation of (85a): A solution of 85-2 (0.16 g; 0.16 mmol)
in 80% aq. HCOOH (5 mL) was stirred at R.T. for 3 h. The solvent
was evaporated and then co-evaporated with toluene. Purification on
a silica column (10 g column) with CH.sub.2Cl.sub.2/MeOH (4-10%
gradient) gave 85a (43 mg, 74%). .sup.1H-NMR (DMSO-d.sub.6):
.delta. 7.75 (d, 1H), 7.33 (d, 2H), 6.07 (dd, 1H), 5.75 (d, 1H),
5.55 (dd, 1H), 5.43 (dt, 1H), 5.43 (t, 1H), 3.79 (dd, 2H), 3.63
(ddd, 2H), 2.64 (sept, 1H), 1.12 (d, 6H). MS: m/z=362.1 [M+1]
Example 83
Preparation of Compound (86a)
##STR00275##
[0624] Preparation of (86-2): 86-2 was prepared using a similar
procedure for preparing 85-2 with the following: 86-1 (220 mg; 0.22
mmol), (2.5 mL), isobutyric anhydride (0.13 mL; 3.6 equiv), EtOAc
(30 mL), and hexanes/EtOAc (30 to 100% gradient) to give 86-2 (175
mg, 85%).
[0625] Preparation of (86a): 86a was prepared using a similar
procedure for preparing 85a with the following: 86-2 (117 mg; 0.13
mmol), 80% aq. HCOOH (4 mL) and CH.sub.2Cl.sub.2/MeOH (4-10%
gradient) to give 86a (36 mg, 77%). .sup.1H-NMR (DMSO-d.sub.6):
.delta. 7.58 (d, 1H), 7.29 (d, 2H), 6.00 (s, 1H), 5.73 (d, 1H),
5.24 (ddd, 1H), 4.55 (dd, 1H), 4.22 (dd, 2H), 3.80 (dd, 2H), 2.58
(sept, 1H), 1.08, 1.07 (2d, 6H). MS: m/z=364 [M+1].
Example 84
Preparation of Compounds (87a)
##STR00276##
[0627] Preparation of (87-2): 87-2 was prepared using a similar
procedure for preparing 46-2 with the following: 87-1 (178 mg, 0.3
mmol), hexanoic anhydride (0.14 mL, 2 equiv.), pyridine (3 mL) to
give 87-2. (120 mg, 50%).
[0628] Preparation of (87a): 87a was prepared using a similar
procedure for preparing 85a with the following: 87-2 (120 mg, 0.15
mmol), 80% aq. HCOOH and CH.sub.2Cl.sub.2/MeOH (4-10% gradient) to
give 87a (62 mg, 85%). .sup.1H-NMR (CDCl.sub.3): .delta. 8.2 (br,
1H), 7.42 (d, 1H), 6.8 (br, 1H), 6.03 (d, 1H), 5.77 (dd, 1H), 5.64
(dd, 1H), 5.51 (ddd, 1H), 4.43 (dd, 2H), 3.82 (dd, 2H), 2.41 (m,
2H), 2.33 (m, 2H), 1.64 (m, 4H), 1.31 (m, 8H), 0.82 (m, 6H). MS:
m/z=488 [M-1].
Example 85
Preparation of Compound (88a)
##STR00277##
[0630] Preparation of (88-2): 88-2 was prepared using a similar
procedure for preparing 85-2 with the following: 85-1 (220 mg; 0.24
mmol), pyridine (3 mL), dodecanoyc anhydride (0.12 g; 1.3 equiv),
EtOAc (50 mL) and hexanes/EtOAc (25 to 80% gradient) to give 88-2
(0.22 g, 85%).
[0631] Preparation of (88a): 88a was prepared using a similar
procedure for preparing 85a with the following: 88-2 (0.19 g; 0.17
mmol), 80% aq. HCOOH (5 mL) and CH.sub.2Cl.sub.2/MeOH (4-10%
gradient) to give 88a (66 mg, 82%). .sup.1H-NMR (DMSO-d.sub.6):
.delta. 7.77 (d, 1H), 7.35 (d, 2H), 6.07 (dd, 1H), 5.77 (d, 1H),
5.60 (dd, 1H), 5.55 (ddd, 1H), 5.43 (t, 1H), 3.78 (dd, 2H), 3.65
(ddd, 2H), 2.41 (m, 2H), 1.56 (m, 2H), 1.24 (m, 16H), 0.85 (m, 3H).
MS: m/z=474 [M-1].
Example 86
Preparation of Compounds (89a) and (90a)
##STR00278##
[0633] Preparation of (89-2): To a solution of 89-1 (175 mg; 0.18
mmol) in MeCN (2.5 mL) at 0.degree. C. was added TMSBr (0.28 mL; 10
equiv.). The mixture was stirred at R.T. for 1 h, evaporated and
treated with water. The obtained white solid was filtered, dried
and washed with CH.sub.2Cl.sub.2. The white solid was then
dissolved in NMP (2 mL) and treated with DIPEA (94 .mu.L; 3 equiv.)
and pivaloyloxymethyliodide (84 .mu.L; 3 equiv.). The mixture was
stirred at R.T. for 1 day, and then partitioned between water (20
mL) and tert-butyl methyl ether (TBME; 60 mL). The organic layer
was washed with saturated aqueous NaHCO.sub.3, water and brine. The
combined aqueous washings were back extracted with TBME (2.times.20
mL). The combined organic extract was dried and purified on a
silica column (10 g column) with CH.sub.2Cl.sub.2/i-PrOH (2-10%
gradient) to give 89-2 (42 mg, 26%).
[0634] Preparation of (89a): A solution of 89-2 in 80% aq. HCOOH
was stirred at R.T. for 3 h. The solvent was evaporated and then
co-evaporated with toluene. Purification on a silica column (10 g
column) with CH.sub.2Cl.sub.2/MeOH (4-15% gradient) gave 89a (17
mg, 74%). .sup.1H-NMR (CD.sub.3OD): .delta. 7.47 (d, 1H), 6.28 (dd,
1H), 6.04 (dd, 1H), 5.77-5.71 (m, 2H), 5.53 (m, 4H), 5.18 (ddd,
1H), 5.60 (dd, 1H), 3.77 (dd, 2H), 1.08 (m, 18H). .sup.31P-NMR
(CD.sub.3OD): .delta. 17.64. MS: m/z=598 [M+1].
[0635] Preparation of (9a): A mixture of 89a (12 mg; 0.02 mmol) in
EtOH (1 mL) and Pd/C (10%; 2.5 mg) was stirred overnight under an
atmospheric pressure of hydrogen. The mixture was filtered through
a Celite pad. The solvent was evaporated and the product was
purified on a silica column (10 g column) with
CH.sub.2Cl.sub.2/MeOH (4-17% gradient) to give 90a (6 mg, 50%).
.sup.1H-NMR (CD.sub.3OD): .delta. 7.51 (d, 1H), 5.79 (d, 1H),
5.65-5.54 (m, 5H), 5.20 (ddd, 1H), 5.60 (dd, 1H), 3.70 (dd, 2H),
2.17-2.06 (m, 1H), 2.02-1.87 (m, 3H), 1.13 (m, 18H). .sup.31P-NMR
(CD.sub.3OD): .delta. 33.16. MS: m/z=600 [M+1].
Example 87
Preparation of Compound (91a)
##STR00279##
[0637] Preparation of (91-2): To a solution of triethylammonium
bis(isopropyloxycarbonyloxymethyl)phosphate (0.33 mmol, prepared
from 110 mg of bis(POC)phosphate and 0.1 mL of Et.sub.3N) in THF (2
mL) was added 86-1 (100 mg; 0.11 mmol), followed by
diisopropylethyl amine (0.19 mL; 10 equiv), BOP-Cl (140 mg; 5
equiv) and 3-nitro-1,2,4-triazole (63 mg; 5 equiv). The mixture was
stirred at R.T. for 90 mins., and then diluted with
CH.sub.2Cl.sub.2 (30 mL). The mixture was washed with saturated
aqueous NaHCO.sub.3 and brine. The mixture was dried with
Na.sub.2SO.sub.4. The solvent was evaporated, and the residue was
purified on a silica column (10 g column) with hexanes/EtOAc
(40-100% gradient) to give 91-2 (117 mg, 90%).
[0638] Preparation of (91a): 91a was prepared using a similar
procedure for preparing 85a with the following: 91-2 (87 mg; 0.07
mmol), 80% aq. HCOOH (5 mL) and CH.sub.2Cl.sub.2/MeOH (4-15%
gradient) to give 91a (36 mg, 85%). .sup.1H-NMR (CD.sub.3CN):
.delta. 7.67 (dd, 1H), 6.35 (dd, 1H), 6.1 (br, 2H), 5.82 (d, 1H),
5.62 (m, 4H), 5.22 (dm, 1H), 4.98 (br, 1H), 4.89 (m, 2H), 4.49 (d,
1H), 4.34 (m, 2H), 3.88 (dd, 2H), 1.29 (d, 6H), 1.28 (d, 6H);
.sup.31P-NMR (CD.sub.3CN): .delta. -4.49. MS: m/z=606 [M+1].
Example 88
Preparation of Compound (92a)
##STR00280##
[0640] Preparation of (92-2) and (92-3): To a solution of
triethylammonium bis(POM)phosphate (0.48 mmol, prepared from 176 mg
of bis(POM)phosphate and 0.15 mL of Et.sub.3N) in THF (2 mL) was
added 92-1 (150 mg; 0.18 mmol) followed by diisopropylethyl amine
(0.31 mL; 10 equiv), BOP-Cl (229 mg; 5 equiv), and
3-nitro-1,2,4-triazole (103 mg; 5 equiv). The mixture was stirred
at R.T. for 90 mins., and then diluted with CH.sub.2Cl.sub.2 (30
mL). The mixture was washed with saturated aqueous NaHCO.sub.3 and
brine. The mixture was dried with Na.sub.2SO.sub.4. The solvent was
evaporated, and the residue was purified on a silica column (10 g
column) with CH.sub.2Cl.sub.2/i-PrOH (2-10% gradient) to obtain
92-2 (44 mg, 21%) and 92-3 (73 mg, 28%).
[0641] Preparation of (92a): A mixture of 92-2 and 92-3 (73 mg and
44 mg) and 80% aq. HCOOH (3 mL) was heated for 30 mins., at
35.degree. C. The solvent was evaporated and then coevaporated with
toluene. The solvent was evaporated, and the residue was purified
on a silica column (10 g column) with CH.sub.2Cl.sub.2/MeOH (4-10%
gradient) to obtain 92a (40 mg, 75%). .sup.1H-NMR (DMSO-D.sub.6):
.delta. 10.6 (br, 1H), 7.76 (s, 1H), 6.44 (br, 2H), 5.99 (dd, 1H),
5.83 (d, 1H), 5.53-5.27 (2 m, 6H), 4.39 (dt, 1H), 4.04 (m, 2H),
1.17 (s, 3H), 1.06, 1.08 (2 s, 18H). .sup.31P-NMR (DMSO-d.sub.6):
.delta. -4.09. MS: m/z=608 [M+1].
Example 89
Preparation of Compound (93a)
##STR00281##
[0643] Preparation of (93-2) and (93-3): 93-2 and 93-3 (68 mg and
80 mg, respectively) were prepared in the same manner from 93-1
(200 mg; 0.23 mmol) and bis(POM) phosphate (230 mg) with DIPEA (0.4
mL), BOPCl (290 mg), and 3-nitro-1,2,4-triazole (130 mg) in THF (3
mL) as 92-2 and 92-3 from 92-1.
[0644] Preparation of (93a): 93-2 and 93-3 (68 mg and 80 mg,
respectively) were converted into 93 (42 mg) with formic acid in
the same manner as 92 from 92-2 and 92-3. .sup.1H-NMR
(DMSO-D.sub.6): .delta. 7.73 (s, 1H), 6.46 (br, 2H), 6.04 (dd, 1H),
5.91 (dd, 1H), 5.87 (d, 1H), 5.48 (d, 4H), 5.33 (m, 1H), 5.24 (ddd,
1H), 4.60 (dt, 1H), 4.07 (m, 2H), 1.07, 1.06, 1.05 (4 s, 18H).
.sup.31P-NMR (DMSO-d.sub.6): .delta. -4.37. MS: m/z=620 [M+1].
Example 90
Preparation of Compound (94a)
##STR00282##
[0646] To a solution of 93a (53 mg; 0.09 mmol) in EtOH (2 mL) was
added 10% Pd/C (10 mg). The mixture stirred under hydrogen at
atmospheric pressure for 1 h. The mixture was filtered through a
Celite pad, and the filtrate evaporated. Purification on a silica
column (10 g column) with CH.sub.2Cl.sub.2/MeOH (4-11% gradient)
yielded 94a (45 mg, 81%). .sup.1H-NMR (DMSO-D.sub.6): .delta. 10.6
(br, 1H), 7.81 (s, 1H), 6.4 (br, 2H), 5.97 (dd, 1H), 5.85 (d, 1H),
5.60-5.44 (m, 5H), 4.37 (m, 1H), 4.11 (ddd, 2H), 1.66 (m, 2H),
1.09, 1.06 (2 s, 18H), 0.81 (7, 3H); .sup.31P-NMR (DMSO-d.sub.6):
.delta. -4.10. MS: m/z=622 [M+1].
Example 91
Preparation of Compounds (95a) and (96a)
##STR00283## ##STR00284##
[0648] Preparation of (95-1): To a solution of
5-Amino-2H-[1,2,4]triazin-3-one (180 mg, 1.5 mmol) in HMDS was
added a catalytic amount of (NH.sub.4).sub.4SO.sub.4. The mixture
was heated to reflux for 5 h. HMDS was evaporated to give a crude
product. To a solution of the crude product in anhydrous CH.sub.3CN
was added 70a (220 mg, 0.5 mmol) and TMSOTf (0.45 mL, 2.5 mmol).
The mixture was heated to reflux for 24 h in a sealed tube. The
reaction was quenched with NaHCO.sub.3 and diluted with EA. The
organic solvent was removed, and the residue was purified by
prep-TLC first, and the by RP-HPLC (0.5% HCOOH in water and MeCN)
to give the pure 95-1 (100 mg, 46%).
[0649] Preparation of (95-2): To a solution of 95-1 (80 mg, 0.18
mmol) in anhydrous CH.sub.3CN was added 1,2,4-triazole (911 mg,
11.7 mmol) and TEA (1.45 g, 14.4 mmol). The mixture was cooled to
0.degree. C. and POCl.sub.3 was added. The reaction mixture was
stirred at 25.degree. C. for 24 h. The solvent was evaporated and
partitioned with EA and water. The organic layer was concentrated
to give the crude 95-2 (80 mg, 90%).
[0650] Preparation of (95a): 95-2 (90 mg, 0.18 mmol) was dissolved
in 20 mL of saturated THF ammonia. The resulting solution was
stirred at 25.degree. C. for 2 h. The solvent was removed, and the
residue was purified on a silica gel column (EA:PE=6:1) to give 95a
as a white solid (70 mg, 70%).
[0651] Preparation of (96a): 95a (70 mg, 0.16 mmol) was dissolved
in 20 mL of saturated MeOH ammonia. The resulting solution was
stirred at 25.degree. C. for 2 h. The solvent was removed, and the
residue was purified by RP-HPLC (0.5% HCOOH in water and MeCN) to
give 96a (5 mg, 11%) as a white solid. .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 7.57 (s, 1H), 6.35 (dd, J=3.6 Hz, J=15.6 Hz, 1H),
5.45-5.47 (m, 1H), 4.70 (dd, J=4.8 Hz, J=16.2 Hz, 1H), 3.83 (s,
2H), 3.71 (d, J=1.6 Hz, 2H). ESI-TOF-MS: m/z 295.1 [M+H].sup.+.
Example 92
Preparation of Compounds (97a-g)
##STR00285##
[0653] Dry nucleoside (0.05 mmol) was dissolved in a mixture of DMF
(3 mL) and DMA-DMF (0.04 mL, 0.1 mmol). The reaction was kept at
ambient temperature for 4 h and then evaporated to dryness. The
residue was dissolved in a mixture of PO(OMe).sub.3 (0.7 mL) and
pyridine (0.3 mL). The mixture was evaporated in vacuum for 15 min.
at 42.degree. C., than cooled down to R.T. N-Methylimidazole (0.009
mL, 0.11 mmol) was added followed by POCl.sub.3 (9 .mu.l, 0.11
mmol). The mixture was kept at R.T. for 20-40 mins. The reaction
was controlled by LCMS and monitored by the appearance of the
corresponding nucleoside 5'-monophosphate. After completion of the
reaction, tetrabutylammonium salt of pyrophosphate (150 mg) was
added, followed by DMF (0.5 mL) to get a homogeneous solution.
After 1.5 h at ambient temperature, the reaction was diluted with
water (10 mL). The mixture was loaded on the column HiLoad 16/10
with Q Sepharose High Performance, and separation was done in a
linear gradient of NaCl from 0 to 1N in 50 mM TRIS-buffer (pH7.5).
The triphosphate (97a-f) was eluted at 75-80% B. The corresponding
fractions were concentrated. The residue was dissolved in 5%
ammonium hydroxide, kept for 15 min. at R.T. and concentrated.
Desalting was achieved by RP HPLC on Synergy 4 micron Hydro-RP
column (Phenominex). A linear gradient of methanol from 0 to 30% in
50 mM triethylammonium acetate buffer (pH 7.5) was used for
elution. The corresponding fractions were combined, concentrated
and lyophilized 3 times to remove excess of buffer.
TABLE-US-00004 TABLE 4 Triphosphates obtained from Example 92
Structure MS (M-1) P(.alpha.) P(.beta.) P(.gamma.) ##STR00286##
528.0 -6.71 -6.82 (d) -21.43 (t) -11.35 -11.47 (d) ##STR00287##
544.0 -6.25 (bs) -21.45 (bs) -11.44 -11.56 (d) ##STR00288## 575.7
-8.86 -9.00 (d) -22.95 (t) -11.81 -11.94 (d) ##STR00289## 545.9
-9.41 -9.44 (d) -23.04 (t) -12.00 -12.13 (d) ##STR00290## 552.1
-10.32 -10.44 (d) -23.26 (t) -11.84 -11.96 (d) ##STR00291## 508.4
-8.30 (bs) -22.72 (bs) -11.51 -11.63 (d) ##STR00292## 550.1 -9.17
-9.29 (d) -23.04 (t) -11.97 -12.09 (d)
Example 93
Preparation of Compounds (98a-e) and (99a)
[0654] Dry nucleoside (0.05 mmol) was dissolved in a mixture of
PO(OMe).sub.3 (0.7 mL) and pyridine (0.3 mL). The mixture was
evaporated in vacuum for 15 mins. at 42.degree. C., than cooled
down to R.T. N-Methylimidazole (0.009 mL, 0.11 mmol) was added
followed by POCl.sub.3 (9 .mu.l, 0.11 mmol). The mixture was kept
at R.T. for 20-40 mins. The reaction was controlled by LCMS and
monitored by the appearance of the corresponding nucleoside
5'-monophosphate. After completion of the reaction,
tetrabutylammonium salt of pyrophosphate (150 mg) was added,
followed by DMF (0.5 mL) to get a homogeneous solution. After 1.5 h
at ambient temperature, the reaction was diluted with water (10 mL)
and loaded on the column HiLoad 16/10 with Q Sepharose High
Performance. Separation was done in a linear gradient of NaCl from
0 to 1N in 50 mM TRIS-buffer (pH7.5). The triphosphate (98a-e) was
eluted at 75-80% B. The corresponding fractions were concentrated.
Desalting was achieved by RP HPLC on Synergy 4 micron Hydro-RP
column (Phenominex). A linear gradient of methanol from 0 to 30% in
50 mM triethylammonium acetate buffer (pH 7.5) was used for
elution. The corresponding fractions were combined, concentrated
and lyophilized 3 times to remove excess of buffer.
TABLE-US-00005 TABLE 5 Compounds obtained from Example 93 Structure
MS (M-1) P(.alpha.) P(.beta.) P(.gamma.) ##STR00293## 538.0 -5.21
-5.33 (d) -20.56 (t) -11.09 -11.20 (t) ##STR00294## 556.2 -10.85
(bs) -23.11 (bs) -11.76 -11.88 (d) ##STR00295## 540.4 -8.86 (bs)
-23.84 (t) -11.68 -11.80(d) ##STR00296## 536.0 -9.35 -9.47 (d)
-23.05 (t) -11.60 -11.72 (d) ##STR00297## 545.9 -10.54 -10.66
-23.26 -11.80 -11.93 (d) ##STR00298## 357.2 1.42 (s) NA NA
Example 94
Preparation of Compound (100a)
##STR00299##
[0656] Preparation of (100-2): To an ice-cold solution of 100-1 (22
mg; 0.055 mmol) in acetonitrile (0.5 mL) was added TMSBr (80 .mu.L;
10 equiv.). The resulting mixture was stirred at R.T. for 1 h. The
mixture was concentrated, and the residue was partitioned between
water and diethyl ether. The aqueous layer was washed with
Et.sub.2O, neutralized with triethylammonium bicarbonate buffer and
lyophilized to yield the triethylammonium salt of 100-2.
[0657] Preparation of (100a): 100-2 was rendered anhydrous by
coevaporating with pyridine and toluene. Anhydrous 100-2 was
dissolved in HMPA (1 mL) and 1,1-carbonyldiimidazole (32 mg; 0.2
mmol) was added. The mixture was stirred at R.T. for 6 h. A
solution of tetrabutylammonium pyrophosphate (0.22 g; .about.0.2
mmol) in DMF (2 mL) was added. The mixture was stirred overnight at
R.T. The mixture was diluted with triethylammonium acetate buffer
and purified by RP-HPLC with a gradient 0-60% B (A: 50 mM aqueous
TEAA, B: 50 mM TEAA in MeOH) and repurified by RP-HPLC with a
gradient 0-30% B to give 100a. .sup.31P-NMR (D.sub.2O): .delta.
3.22 (d, 1P), -8.21 (br, 1P), -22.91 (br, 1P). MS: m/z=528
(M-1).
Example 95
Preparation of Compound (100b)
##STR00300##
[0659] Preparation of (100-4): 100-4 was prepared from 100-3 (54
mg; 0.13 mmol) in acetonitrile (1.3 mL) with TMSBr (0.18 mL) using
a similar procedure as described for the preparation of 100-2.
[0660] Preparation of (100b): 100b was prepared from 100-4 in HMPA
(2 mL) with CDI (84 mg) and tetrabutylammonium pyrophosphate (0.5
g) in DMF (2 mL) using a similar procedure as described for the
preparation of 100a. .sup.31P-NMR (D.sub.2O): .delta. 17.90 (d,
1P), -9.00 (d, 1P), -22.91 (t, 1P). MS: m/z=530 (M-1).
Example 96
Preparation of Compound (100c)
##STR00301##
[0662] Preparation of (100-6): 100-6 was prepared from 100-5 (40
mg; 0.09 mmol) in acetonitrile (1 mL) with TMSBr (0.1 mL) using a
similar procedure as described for the preparation of 100-2.
[0663] Preparation of (100c): 100c was prepared from 100-6 in HMPA
(1.5 mL) with CDI (50 mg) and tetrabutylammonium pyrophosphate (0.3
g) using a similar procedure as described for the preparation of
100a. .sup.31P-NMR (D.sub.2O): .delta. -7.13 (br, 1P), -10.14 (d,
1P), -22.84 (br, 1P). .sup.19F-NMR (D.sub.2O): .delta. -117.53 (dd,
1F), -197.8 (m, 1F). MS: m/z=545.5 (M-1).
Example 97
Preparation of Compounds (100d) and (100e)
##STR00302##
[0665] Preparation of (100-8): To an ice-cold solution of
diastereomers 100-7 (35 mg; 0.08 mmol) in acetonitrile (1 mL) was
added TMSBr (0.1 mL; 10 equiv.). The resulting mixture was stirred
overnight at R.T. and then concentrated. The residue was
partitioned between water and CH.sub.2Cl.sub.2. The aqueous layer
was washed with CH.sub.2Cl.sub.2, neutralized with triethylammonium
bicarbonate buffer and lyophilized to yield the triethylammonium
salt of 100-8.
[0666] Preparation of (100d) and (100e): 100-8 was rendered
anhydrous by coevaporating with pyridine and toluene. Anhydrous
100-8 was dissolved in DMF (1.5 mL) and CDI (54 mg; 0.3 mmol) was
added. The mixture was stirred at R.T. for 7 h. A solution of
tetrabutylammonium pyrophosphate (0.3 g; .about.0.3 mmol) in DMF (4
mL) was added. The mixture was stirred at R.T for 3 days. The
mixture was diluted with triethylammonium acetate buffer. Two
consecutive RP-HPLC purifications with a gradient 0-60% B (A: 50 mM
aqueous TEAA, B: 50 mM TEAA in MeOH) and 0-40% B gave 100d and 100e
as single diastereomers. 100d: .sup.31P-NMR (D.sub.2O): .delta.
4.28 (dd, 1P), -6.37 (d, 1P), -22.36 (t, 1P). MS: m/z=548.1 (M-1).
100e: .sup.31P-NMR (D.sub.2O): .delta. 4.13 (dd, 1P), -6.38 (d,
1P), -22.46 (t, 1P). MS: m/z=548.1 (M-1).
Example 98
Preparation of Compound (101a)
##STR00303##
[0668] Preparation of (101-1): To a solution of 59-4 (1.5 g, 2.39
mmol) in anhydrous DCM (100 mL) was added Dess-Martin periodinane
(5.2 g, 11.95 mmol) at 0.degree. C. under nitrogen. The mixture was
stirred at R.T. for 5 h. The mixture was poured into NaHCO.sub.3
and Na.sub.2S.sub.2O.sub.3 aq. Solution. The organic layer was
washed with brine, dried over with anhydrous Na.sub.2SO.sub.4, and
concentrated to dryness to give the crude 101-1 (1.5 g) as a white
solid, which was used for the next step without further
purification.
[0669] Preparation of (101-2): To a mixture of
bromo(isobutyl)triphenylphosphorane (4.8 g, 12.03 mmol) in
anhydrous THF (8 mL) was added t-BuOK (11.2 mL, 11.2 mmol) at
0.degree. C. under nitrogen. The mixture was stirred at R.T. for 1
h. A solution of 101-1 (1.0 g, 1.6 mmol) in anhydrous THF (4 mL)
was added dropwise at 0.degree. C. The mixture was stirred at R.T.
for 3 h. The reaction was quenched with a NH.sub.4Cl aq. solution
and extracted with DCM. The organic layer was dried and
concentrated to give a residue, which was purified by silica gel
column chromatography (5% EtOAc in PE) to give 101-2 (793 mg,
74.4%) as a white solid.
[0670] Preparation of (101-3): To a solution of 101-2 (364 mg,
0.547 mmol) in anhydrous CH.sub.3CN (6 mL) were added TPSCl (414
mg, 1.37 mmol), DMAP (167 mg, 1.37 mmol) and NEt.sub.3 (138 mg,
1.37 mmol) at R.T. The mixture was stirred at R.T. for 2 h.
NH.sub.4H (6 mL) was added, and the mixture was stirred for another
1 h. The mixture was diluted with DCM and washed with a NaHCO.sub.3
aq. solution. The organic layer was separated and concentrated to
give a residue, which was purified by silica gel column
chromatography (2% MeOH in DCM) to give 101-3 (347 mg, 95.0%) as
white solid.
[0671] Preparation of (101a): To a solution of 27-3 (347 mg, 0.52
mmol) in MeOH (10 mL) was added NH.sub.4F (1.5 g) at R.T. The
reaction mixture was refluxed for 12 h, and then filtered. The
filtrate was concentrated in vacuo, and the residue was purified by
silica gel column chromatography (10% MeOH in DCM) to give 101a (87
mg, 53%) as a white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz)
.delta. 8.11 (d, J=7.6 Hz, 1H), 6.03 (dd, J=1.2, 17.6 Hz, 1H), 5.88
(d, J=7.2 Hz, 1H), 6.03 (dd, J=1.6, 11.6 Hz, 1H), 5.39 (d, J=10.8
Hz, 1H), 4.88 (dd, J=3.2, 60.0 Hz, 1H), 4.41 (dd, J=4.8, 24.4 Hz,
1H), 3.70 (d, J=12.4 Hz, 1H), 3.57 (d, J=12.0 Hz, 1H), 3.08-3.14
(m, 1H), 0.94-0.98 (m, 6H). ESI-MS: m/z 626.9 [2M+H].sup.+.
Example 99
Preparation of Compound (102a)
##STR00304##
[0673] Preparation of (102-1): To a solution of 101-2 (1.0 g, 1.5
mmol) in MeOH (20 mL) was added NH.sub.4F (6 g) at R.T., and the
mixture was refluxed overnight. After cooling to R.T., the mixture
was filtered, and the filtrate was concentrated. The residue was
purified by silica gel column chromatography (8% MeOH in DCM) to
give 102-1 (400 mg, 85%) as a white solid.
[0674] Preparation of (102-2): To a solution of 102-1 (400 mg, 1.27
mmol) in MeOH (10 mL) was added Pd/C (400 mg) at R.T. The mixture
was stirred at R.T. under a balloon of H.sub.2 for 1.5 h. The
mixture was filtered, and the filtrate was concentrated in vacuo to
give 102-2 (400 mg, 99%) as a white solid.
[0675] Preparation of (102-3): To a solution of 102-2 (400 mg, 1.26
mmol) in anhydrous DMF (5 mL) were added imidazole (968 mg, 14.2
mmol), and TBSCl (1.5 g, 10.0 mmol) at R.T. The mixture was stirred
at 50.degree. C. overnight. The mixture was diluted with DCM and
washed with a NaHCO.sub.3 aq. solution. The organic layer was dried
and concentrated. The residue was purified by silica gel column
chromatography (10% EA in PE) to give 102-3 (676 mg, 98%) as a
white solid.
[0676] Preparation of (102-4): To a solution of 102-3 (676 mg, 1.24
mmol) in anhydrous CH.sub.3CN (6 mL) were added TPSCl (941 mg,
13.11 mmol), DMAP (379 mg, 3.11 mmol) and NEt.sub.3 (314 mg, 3.11
mmol) at R.T. The reaction was stirred at R.T. for 3 h. NH.sub.4OH
(1 mL) was added, and the reaction was stirred for 4 h. The mixture
was diluted with DCM and washed with a NaHCO.sub.3 solution. The
organic layer was dried and concentrated. The residue was purified
by silica gel column chromatography (2% MeOH in DCM) to give 102-4
(450 mg, 67%) as a white solid.
[0677] Preparation of (102a): To a solution of 102-4 (450 mg, 0.83
mmol) in MeOH (10 mL) was added NH.sub.4F (2 g) at R.T. The
reaction mixture was refluxed overnight. After cooling to R.T., the
mixture was filtered, and the filtrate was concentrated. The
residue was purified by silica gel column chromatography (8% MeOH
in DCM) to give 102a (166.6 mg, 64%) as a white solid. .sup.1H NMR
(CD.sub.3OD, 400 MHz) .delta. 8.09 (d, J=7.6 Hz, 1H), 6.07 (d,
J=3.6 Hz, 1H), 6.05 (d, J=2.8 Hz, 1H), 5.89 (d, J=7.6 Hz, 1H), 5.03
(dd, J=5.2, 57.2 Hz, 1H), 4.41 (dd, J=4.2, 17.2 Hz, 1H), 3.74 (d,
J=12.0 Hz, 1H), 3.54 (d, J=12.0 Hz, 1H), 1.23-1.78 (m, 5H), 0.90
(d, J=6.4 Hz, 6H). ESI-MS: m/z 631.1 [2M+H].sup.+.
Example 100
Preparation of Compound (103a)
##STR00305##
[0679] Preparation of (103-2): 103-1 (3.8 g, 6.9 mmol) in 80% AcOH
aq. was stirred at 50.degree. C. for 4 h. The mixture was
concentrated to give a residue, which was purified by silica gel
column chromatography (5% MeOH in DCM) to give the uridine
derivative (1.5 g, 78.2%) as a white solid. To a solution of the
uridine derivative (1.5 g, 5.4 mmol) in Py (10 mL) was added
Ac.sub.2O (1.38 g, 13.5 mmol) at R.T. The mixture was stirred at
R.T. for 12 h. The mixture was concentrated to give a residue,
which was purified by silica gel column chromatography (20% EA in
PE) to give 103-2 (1.3 g, 68%) as a white solid.
[0680] Preparation of (103-3): To a solution of
N-(5-fluoro-2-hydroxy-1,2-dihydropyrimidin-4-yl)benzamide (0.5 g,
2.1 mmol) in anhydrous PhCl (5 mL) was added ammonium sulfate (6
mg, 0.043 mmol), followed by HMDS (0.7 g, 4.3 mmol). The mixture
was heated to 130.degree. C. for 8 h. The mixture was concentrated
under vacuum to 2 mL, and then cooled to 0.degree. C. TMSOTf (310
mg, 1.4 mmol) was then added. After stirring for 10 min at
0.degree. C., 103-2 (150 mg, 0.4 mmol) in PhCl (5 mL) was added.
The mixture was stirred at 130.degree. C. for 10 h. The mixture was
concentrated, and the residue was re-dissolved in DCM (10 mL),
washed with water (5 mL) and saturated NaHCO.sub.3. The organic
layer was dried over Na.sub.2SO.sub.4, evaporated to dryness and
the crude product was purified by silica gel column chromatography
(60% PE in EA) to give 103-3 (30 mg, 16%) as a white solid.
[0681] Preparation of (103a): A solution of 103-3 (150 mg, 0.34
mmol) in NH.sub.3/MeOH (10 mL) was stirred at R.T. for 3 h. The
mixture was concentrated, and the residue was purified by HPLC
separation (0.1% HCOOH in water and MeCN) to give 103a (60 mg, 60%)
as a white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 8.28
(d, J=6.8 Hz, 1H), 6.10 (dd, J=2.0, 15.2 Hz, 1H), 4.99-5.15 (m,
1H), 4.62-4.65 (m, 1H), 4.49-4.55 (m, 2H), 3.89 (dd, J=1.6, 12.0
Hz, 1H), 3.75 (dd, J=1.2, 12.0 Hz, 1H). ESI-MS: m/z 613.1
[2M+Na].sup.+.
Example 101
Preparation of Compound (104a)
##STR00306##
[0683] Preparation of (104-1): 103-3 (150 mg, 0.31 mmol) was
dissolved in 80% aqueous acetic acid (3 mL). The solution was
heated to reflux for 2 h. The mixture was cooled to ambient
temperature and diluted with water (5 mL), neutralized to pH>7
with saturated NaHCO.sub.3 and extracted with EA. The organic layer
was dried and evaporated to dryness. The residue was purified by
silica gel column chromatography (50% EA in PE) to give 104-1 (80
mg, 70%) as a white solid.
[0684] Preparation of (104a): 104-1 (80 mg, 0.22 mmol) in saturated
NH.sub.3/MeOH (10 mL) was stirred at R.T. for 3 h. The mixture was
concentrated, and the residue was purified by silica gel column
chromatography (5% MeOH in DCM) to give 104a (40 mg, 60%) as a
white solid. .sup.1H NMR (CD.sub.3OD, 400 MHz) .delta. 58.30 (d,
J=6.8 Hz, 1H), 6.18 (dd, J=4.0, 14.0 Hz, 1H), 5.13-5.65 (m, 1H),
4.52-4.56 (m, 1H), 3.980-3.95 (m, 2H), 3.76 (s, 3H). ESI-MS: nm/z
319.1 [M+Na].sup.+.
Example 102
Preparation of Compound (105a)
##STR00307##
[0686] Preparation of (105-2): To a solution of triethylammonium
bis(isopropyloxycarbonyloxymethyl)phosphate (0.065 mmol, prepared
from 22 mg of bis(POC)phosphate and Et.sub.3N) in THF was added
105-1 (31 mg; 0.05 mmol). The resulting mixture evaporated, and the
residue was rendered anhydrous by coevaporation with pyridine,
followed by toluene. The anhydrous evaporated residue was dissolved
TH (1 mL) and cooled in an ice-bath. To the solution was added
diisopropylethyl amine (35 .mu.L; 4 equiv), followed by BOP-Cl (25
mg; 2 equiv) and 3-nitro-1,2,4-triazole (11 mg; 2 equiv). The
mixture was stirred at 0.degree. C. for 90 min. The mixture was
diluted with CH.sub.2Cl.sub.2, washed with saturated aq.
NaHCO.sub.3 and brine, and dried with Na.sub.2SO.sub.4. The
evaporated residue was purified on silica (10 g column) with a
CH.sub.2Cl.sub.2/i-PrOH solvent system (3-10% gradient) to give
105-2 (13 mg, 28%).
[0687] Preparation of (105a): A solution of 105-2 (13 mg; 0.014
mmol) in 80% aq. HCOOH (2 mL) was stirred at R. T. for 3 h. The
mixture was evaporated and then coevaporated with toluene. The
product was purified on silica (10 g column) with a
CH.sub.2Cl.sub.2/MeOH solvent system (4-15% gradient) to give 105a
(7 mg, 78%). .sup.1H-NMR (DMSO-d.sub.6): .delta. 7.52 (d, 1H),
7.28, 7.24 (2br s, 2H) 5.92 (dd, 1H), 5.74 (d, 1H), 5.69 (d, 1H),
5.62 (d, 4H), 4.97 (ddd, 1H), 4.82 (m, 2H), 4.38 (dt, 1H), 4.07 (m,
2H), 1.23 (m, 12H), 1.04 (m, 1H), 0.37 (n, 4H). .sup.31P-NMR
(DMSO-d.sub.6): .delta. -4.51. .sup.19F-NMR (DMSO-d.sub.6): .delta.
-199.23 (dt). MS: m/z=598.4 (M+1).
Example 103
Preparation of Compound (106a)
##STR00308##
[0689] Preparation of (106-1): 106-1 (15 mg; 30% yield) was
prepared in the same manner from 43-5 (32 mg; 0.057 mmol) and
bis(POC)phosphate (24 mg) with DIPEA (40 .mu.L), BopCl (29 mg) and
3-nitro-1,2,4-triazole (13 mg) as 105-2 from 105-1.
[0690] Preparation of (106a): 106-1 (15 mg) was converted in formic
acid to 106a (8 mg; 78% yield) in the same manner as 105-2 to 105a.
.sup.1H-NMR (DMSO-d.sub.6): .delta. 7.55 (d, 1H), 7.32, 7.27 (2 br
s, 2H) 6.06 (dd, 1H), 5.84 (d, 1H), 5.73 (d, 1H), 5.61 (d, 4H),
5.08 (ddd, 1H), 4.83 (m, 2H), 4.36 (m, 1H), 4.21 (dd, H), 4.16 (dd,
1H), 3.56 (d, 1H), 3.49 (d, 1H), 3.28 (s, 3H), 1.25, 1.24 (2 d,
12H). .sup.31P-NMR (DMSO-d.sub.6): .delta. -4.45. MS: m/z=602.4
(M+1).
Example 104
Preparation of Compound (107a)
##STR00309##
[0692] Preparation of (107-1): 107-1 (30 mg; 30% yield) was
prepared in the same manner from 40-10 (65 mg; 0.115 mmol) and
bis(POC)phosphate (49 mg) with DIPEA (80 .mu.L), BopCl (58 mg) and
3-nitro-1,2,4-triazole (26 mg) as 105-2 from 105-1.
[0693] Preparation of (106a): 107-1 (30 mg) was converted in formic
acid to 107a (15 mg; 73% yield) in the same manner as 105-2 to
105a. .sup.1H-NMR (DMSO-d.sub.6): .delta. 7.60 (d, 1H), 7.36, 7.32
(2 br s, 2H) 6.02 (m, 2H), 5.74 (d, 1H), 5.62 (m, 4H), 5.17 (ddd,
1H), 4.99 (dq, 1H), 4.83 (m, 2H), 4.61 (m, 1H), 4.19 (m, 2H), 1.40
(dd, 3H), 1.24, 1.23 (2 d, 12H). .sup.31P-NMR (DMSO-d.sub.6):
.delta. -4.52. .sup.19F-NMR (DMSO-d.sub.6): .delta. -185.92 (m,
1F), -200.48 (d, 1F). MS: m/z=604.3 (M+1).
Example 105
Preparation of Compound (108a)
##STR00310##
[0695] To a solution of 4'-ethyl-2'-fluorocytidine (50 mg, 0.183
mmol) in DMF (1 mL) were added DCC (113 mg, 0.55 mmol), isobutyric
acid (48.5 .mu.l, 0.55 mmol) and DMAP (22 mg, 0.183 mmol). The
mixture was stirred at R.T. overnight. The mixture was filtered,
and the filtrate was concentrated with a rotary evaporator until
half of its original volume was achieved. EA was added to the
mixture. The mixture was washed with water, followed by brine. The
mixture was dried over anhydrous Na.sub.2SO.sub.4 and concentrated
in vacuo to give a residue, which was purified by silica gel with
DCM/MeOH=95:5 to give 108a (40.8 mg, 54%) as a white solid. .sup.1H
NMR (DMSO-d6, 400 MHz) .delta. 7.67 (d, J=7.2 Hz, 1H), 7.34 (br s,
2H), 5.85, 5.8 (2d, J=21.2, 22 Hz, 1H), 5.72 (d, J=7.6 Hz, 1H),
5.55-5.41 (m, 2H), 4.1 (q, 2H), 2.68-2.52 (m, 2H), 1.77-1.64 (m,
2H), 1.13, 1.14 (2s, 2.times.3H), 1.09-1.07 (m, 6H), 0.96 (t, J=7.6
Hz, 3H); MS/z 414 (M-H.sup.+), 829 (2M+H.sup.+).
Example 106
Preparation of Compound (109a)
##STR00311##
[0697] 3',5'-diacetylnucleoside (36 mg, 1 mmol) was dissolved in
methanol saturated with NH.sub.4OH and kept overnight at R.T. The
solvent was evaporated, and the product isolated by column
chromatography in gradient of methanol in DCM from 0 to 15% on a 10
g Biotage cartridge. The product was 109a obtained (20 mg, 73%).
.sup.1H-NMR (DMSO-d.sub.6): .delta. 11.4 (s, 1H), 11.84-11.82 (d,
1H); 6.10-6.05 (m, 1H), 5.95-5.83 (d, 1H), 5.71 (s, 1H), 5.65-5.63
(d, 1H), 5.37-3.36 (t, 1H), 5.26-5.20 (t, 1H), 5.11-5.07 (t, 1H),
4.56-4.55 (m, 1H), 4.46-4.33 (m, 2H), 3.58-3.56 (m, 2H). MS 277.2
(M-H).
Example 107
Preparation of Compound (110a)
##STR00312##
[0699] Preparation of (110-): To a solution of 70a (6.55 g, 2.1
mmol) and the benzoyl protected base moiety (2.3 g, 5.3 mmol) in
PhCl (50 mL) was added TMSOTf (3.6 g, 16.1 mmol). After addition,
the mixture was heated to 140.degree. C. for 8 h. The mixture was
cooled to R.T., and evaporated to give a residue. The residue was
re-dissolved in DCM and washed with saturated NaHCO.sub.3 and
brine. The organic layer was dried and concentrated to give a
residue, which was purified by silica gel column (40% EA in PE) to
give 110-1 (300 mg, 10%) as a white solid.
[0700] Preparation of (110a): 110-1 (300 mg, 0.55 mmol) in 80%
aqueous acetic acid (5 mL) was heated to reflux for 2 h. The
mixture was cooled to ambient temperature and diluted with water (5
mL), and then extracted with EA. The organic layer was washed with
saturated NaHCO.sub.3 and brine. The mixture was dried and
concentrated to give a residue, which was purified by silica gel
column (10% EA in PE) to give the protected uridine derivative (180
mg, 70%) as a white solid. The protected uridine derivative (180
mg, 0.4 mmol) in saturated NH3/MeOH (10 mL) was stirred at R.T. for
3 h. The mixture was concentrated to give a residue, which was
purified by preparative HPLC (0.1% HCOOH in water and MeCN) to give
110a (80 mg, 60%) as a white solid. .sup.1H NMR (CD.sub.3OD, 400
MHz) .delta. 8.31 (d, J=6.8 Hz, 1H), 6.17 (dd, J=4.0, 14.0 Hz, 1H),
5.13-5.27 (m, 1H), 4.52-4.56 (m, 1H), 3.92 (dd, J=12.0, 58.8 Hz,
2H). ESI-TOF-MS: m/z 334.7 [M+Na].sup.+.
Example 108
RSV Antiviral Assays
[0701] CPE reduction assays are performed as described by Sidwell
and Huffman et al., Appl Microbiol. (1971) 22(5):797-801 with
slight modifications. HEp-2 cells (ATCC) at a concentration of 6000
cell/well are infected with RSV Long strain (ATCC) at a
multiplicity of infection (m.o.i.) of 0.01, and each of the test
compounds are provided to duplicate wells at final concentrations
starting from 100 .mu.M using 1/3 stepwise dilutions. For each
compound, two wells are set aside as uninfected, untreated cell
controls (CC), and two wells per test compound receive virus only
as a control for virus replication (VC). The assay is stopped after
6 days, before all of the cells in the virus-infected untreated
control wells exhibited signs of virus cytopathology (giant cell
formation, syncytia). At the end of the incubation, 20 .mu.l of
cell counting kit-8 reagent (CCK-8, Dojindo Molecular Technologies,
Inc.) are added to each well. After 4 hour incubation, the
absorbance is measured in each well according to manufacturer's
instruction, and the 50% effective concentration (EC.sub.50) is
calculated by using regression analysis, based on the mean O.D. at
each concentration of compound.
[0702] RT-PCR based assays were performed in HEp-2 cells (ATCC:
CCL-23) at a concentration of 20000 cell/well were plated in 96
well plates and incubated overnight. Each of the test compounds
were 1/3 serially diluted and dosed to HEp-2 cells in duplicates.
The highest final concentration for each compound was 100 uM. After
24 hour compound pre-incubation, RSV A2 (ATCC: VR-1540) at MOI of
0.1 was added. Two wells per compound were set aside as uninfected,
untreated cell controls (CC), and four wells per test compound
received virus only as a control for virus replication (VC). The
assay was stopped 4 days after virus infection and conditioned
media was removed for viral RNA isolation. The quantities of the
RSV virus were measured by real-time PCR using a set of RSV
specific primers and probe. The data was analyzed with Prism
software with EC50 defined as drug concentration that reduced the
viral load 50% from the viral control (VC).
[0703] Standard RSV polymerase assays were conducted in the
presence of 3 .mu.L extract of RSV-infected cells in a reaction
buffer containing 50 mM tris-acetate pH 8, 120 mM K-acetate, 4.5 mM
MgCl.sub.2, 5% glycerol, 2 mM EDTA, 50 ug/mL BSA, and 3 mM DTT.
Varying concentration of test compounds were used to initiate RNA
synthesis for 120 mins at 30.degree. C., and radioactive 33P GTP
(15 uCi) was used as tracer. The reaction was stopped by adding 50
mM EDTA, and RNA samples were purified through G-50 size exclusion
spin columns and phenol-chloroform extraction. The radio-labeled
RNA products were resolved by electrophoresis on a 6%
polyacrylamide TBE gel, and visualized and quantitated after being
exposed on a phosphorImager screen. Polymerase inhibition
experiments (IC.sub.50) were conducted the same way in the presence
of increasing concentration of test compounds.
[0704] Compounds of Formula (I), Formula (II) and Formula (III) are
active in the assay as noted in Tables 6 and 7. In Table 6, `A`
indicates an EC.sub.50<2 .mu.M, `B` indicates an EC.sub.50 of
.gtoreq.2 .mu.M and <10 .mu.M and `C` indicates an
EC.sub.50.gtoreq.10 .mu.M and <50 .mu.M. In Table 7, `A`
indicates an EC.sub.50<1 .mu.M, `B` indicates an EC.sub.50 of
.gtoreq.1 .mu.M and <10 .mu.M and `C` indicates an
EC.sub.50.gtoreq.10 .mu.M and <100 .mu.M.
TABLE-US-00006 TABLE 6 Activity of compounds as determined by RSV
polymerase assay No. EC.sub.50 No. EC.sub.50 No. EC.sub.50 No.
EC.sub.50 No. EC.sub.50 35a A 36i B 56c A 97b A 97g A 36a A 36j B
56da A 97c A 98b A 36c A 56a B 56e A 97d A 98c A 36e A 56a B 97a
A
TABLE-US-00007 TABLE 7 Activity of compounds as determined by
RT-PCR assay No. EC.sub.50 No. EC.sub.50 No. EC.sub.50 No.
EC.sub.50 No. EC.sub.50 No. EC.sub.50 1a C 14a A 28a B 48a B 81a B
106a C 2a C 20a B 30a A 50a A 82a A 108a B 3a A 21a A 31a B 52a A
83a B -- -- 4a C 22a C 33a A 58a C 85a A -- -- 7a A 23a A 39a B 69a
A 86a A -- -- 9a C 25a C 41a B 71a A 87a A -- -- 11a B 26a B 46a B
73a C 92a C -- -- 13a C 27a B 45a C 76a A 105a C -- --
Example 109
Influenza Antiviral Assay
[0705] Human lung carcinoma A549 cells (ATCC, Manassas, Va.) were
plated at a density of 5.times.10.sup.4 cells/mL (5.times.10.sup.3
cells/well) in assay media (Ham's F12 media supplemented with 0.3%
FBS, 1% penicillin/streptomycin (all Mediatech, Manassas, Va.) and
1% DMSO (Sigma-Aldrich, St Louis, Mo.)) in black 96-well plates.
After 24 hours, serially diluted test compounds were added to cells
and incubated for an additional 24 hours. Cells were infected with
250 IU/well of Influenza strain A/WSN/33 (H1N1) (Virapur, San Diego
Calif.) and incubated for 20 hours at 37.degree. C., 5% CO.sub.2.
The cell culture supernatant was aspirated off and 50 .mu.L of 25
.mu.M 2'-(4-Methylumbelliferyl)-a-D-N-acetylneuraminic acid
(Sigma-Aldrich) dissolved in 33 mM MES, pH 6.5 (Emerald Biosystems,
Bainbridge Island, Wash.) was added to the cells. After incubation
for 45 mins at 30.degree. C., reactions were stopped by addition of
150 .mu.L stop solution (100 mM glycine, pH 10.5, 25% ethanol, all
Sigma-Aldrich). Fluorescence was measured with excitation and
emission filters of 355 and 460 nm, respectively, on a Victor X3
multi-label plate reader (Perkin Elmer, Waltham, Mass.).
Cytotoxicity of uninfected parallel cultures was determined by
addition of 100 .mu.L of CellTiter-Glo.RTM. reagent (Promega,
Madison, Wis.), and incubation for 10 mins at R.T. Luminescence was
measured on a Victor X3 multi-label plate reader.
[0706] Compounds of Formula (I), Formula (II) and Formula (III) are
active in the assay as noted in Table 8, where `A` indicates an
EC.sub.50<20 .mu.M, `B` indicates an EC.sub.50 of .gtoreq.20
.mu.M and <100 .mu.M and `C` indicates an EC.sub.50.gtoreq.100
.mu.M and <250 .mu.M.
TABLE-US-00008 TABLE 8 Activity of compounds No. % Inhibition 1a C
2a C 3a C 4a C 6a C 7a C 9a C 12a C 16a C 17a C 18a C 20a C 21a C
22a C 23a C 25a A 26a C 27a B 28a C 30a C 31a C 39a B
Example 110
Influenza Pol Assay
[0707] Recombinant influenza polymerase trimer is obtained as
described (Aggarwal S. et al., PLoS ONE 2010). Standard RNA
polymerization assays are conducted in the presence of 0.15 uM
enzyme, 1.5 uM 50-mer oligonucleotide template, 400 uM AG primer
and varying concentration of the test compounds are incubated
together for 40 minutes at 30.degree. C. Radioactive 33P GTP are
used as the tracer and the radio-labeled RNA products are resolved
by electrophoresis on a 15% polyacrylamide TBE gel, and is
visualized and quantitated after being exposed on a phosphorImager
screen. Polymerase inhibition experiments (IC.sub.50) are conducted
the same way in the presence of increasing concentration of test
compounds.
[0708] Although the foregoing has been described in some detail by
way of illustrations and examples for purposes of clarity and
understanding, it will be understood by those of skill in the art
that numerous and various modifications can be made without
departing from the spirit of the present disclosure. Therefore, it
should be clearly understood that the forms disclosed herein are
illustrative only and are not intended to limit the scope of the
present disclosure, but rather to also cover all modification and
alternatives coming with the true scope and spirit of the
invention.
* * * * *