U.S. patent application number 15/446831 was filed with the patent office on 2017-11-23 for tetracycline compounds.
The applicant listed for this patent is Tetraphase Pharmaceuticals,Inc.. Invention is credited to Yonghong Deng, Louis Plamondon, Magnus P. Ronn, Cuixiang Sun, Joyce A. Sutcliffe, Xiao-Yi Xiao, Jingye Zhou.
Application Number | 20170334841 15/446831 |
Document ID | / |
Family ID | 43513614 |
Filed Date | 2017-11-23 |
United States Patent
Application |
20170334841 |
Kind Code |
A1 |
Deng; Yonghong ; et
al. |
November 23, 2017 |
TETRACYCLINE COMPOUNDS
Abstract
The present invention is directed to a compound represented by
Structural Formula (I): ##STR00001## or a pharmaceutically
acceptable salt thereof. The variables for Structural Formula (I)
are defined herein. Also described is a pharmaceutical composition
comprising the compound of Structural Formula (I) and its
therapeutic use.
Inventors: |
Deng; Yonghong; (Watertown,
MA) ; Plamondon; Louis; (Quebec, CA) ; Sun;
Cuixiang; (Arlington, MA) ; Xiao; Xiao-Yi;
(Lexington, MA) ; Zhou; Jingye; (Shanghai, CN)
; Sutcliffe; Joyce A.; (Newton, MA) ; Ronn; Magnus
P.; (Melrose, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Tetraphase Pharmaceuticals,Inc. |
Watertown |
MA |
US |
|
|
Family ID: |
43513614 |
Appl. No.: |
15/446831 |
Filed: |
March 1, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13391407 |
May 2, 2012 |
9624166 |
|
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PCT/US2010/047035 |
Aug 27, 2010 |
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15446831 |
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61275507 |
Aug 28, 2009 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07C 2601/02 20170501;
C07C 311/09 20130101; C07D 213/70 20130101; C07D 233/60 20130101;
C07D 233/84 20130101; C07D 211/60 20130101; C07D 307/64 20130101;
C07C 311/44 20130101; C07C 2601/04 20170501; C07D 209/44 20130101;
C07D 213/82 20130101; A61P 31/00 20180101; C07C 2601/08 20170501;
A61P 31/04 20180101; C07D 205/04 20130101; C07C 311/21 20130101;
C07D 209/52 20130101; C07D 223/06 20130101; C07C 2603/46 20170501;
C07C 2601/14 20170501; C07D 295/15 20130101; C07C 311/29 20130101;
C07C 307/10 20130101; C07C 311/13 20130101; C07D 211/54 20130101;
C07C 237/26 20130101; C07D 207/16 20130101; C07C 2601/16 20170501;
C07D 213/74 20130101; C07C 311/08 20130101; C07C 2603/40 20170501;
C07D 207/12 20130101; C07D 231/18 20130101; C07C 2603/44
20170501 |
International
Class: |
C07C 311/08 20060101
C07C311/08; C07D 295/15 20060101 C07D295/15; C07C 237/26 20060101
C07C237/26; C07C 307/10 20060101 C07C307/10; C07C 311/09 20060101
C07C311/09; C07C 311/13 20060101 C07C311/13; C07C 311/21 20060101
C07C311/21; C07C 311/29 20060101 C07C311/29; C07C 311/44 20060101
C07C311/44; C07D 233/84 20060101 C07D233/84; C07D 233/60 20060101
C07D233/60; C07D 231/18 20060101 C07D231/18; C07D 223/06 20060101
C07D223/06; C07D 213/82 20060101 C07D213/82; C07D 213/74 20060101
C07D213/74; C07D 213/70 20060101 C07D213/70; C07D 211/60 20060101
C07D211/60; C07D 211/54 20060101 C07D211/54; C07D 209/52 20060101
C07D209/52; C07D 209/44 20060101 C07D209/44; C07D 207/16 20060101
C07D207/16; C07D 207/12 20060101 C07D207/12; C07D 205/04 20060101
C07D205/04; C07D 307/64 20060101 C07D307/64 |
Claims
1. A compound of Structural Formula I: ##STR00280## or a
pharmaceutically acceptable salt thereof, wherein: X is selected
from --OCF.sub.3 and --OCH.sub.3; Y is selected from hydrogen,
--(C.sub.1-C.sub.7)alkyl, carbocyclyl,
--(C.sub.1-C.sub.4)alkylene-N(R.sup.2)(R.sup.3),
--(C.sub.1-C.sub.4)alkylene-N(R.sup.F)--C(O)--[C(R.sup.5a)(R.sup.5b)].sub-
.0-4-N(R.sup.2)(R.sup.3), --CH.dbd.N--OR.sup.2,
--C(O)--N(R.sup.2)(R.sup.4), --NO.sub.2, --COOH, --OH,
--N.dbd.CH--N(R.sup.2)(R.sup.3), --N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--[C(R.sup.5a)(R.sup.5b)].sub.1-4--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--(C.sub.1-C.sub.6)alkyl,
--N(R.sup.F)--C(O)-heterocyclyl, --N(R.sup.F)--C(O)-carbocyclyl,
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4)
alkylene-N(R.sup.2)(R.sup.3),
--N(R.sup.F)--S(O).sub.m--N(R.sup.2)(R.sup.4) and
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4)alkylene-carbocyclyl;
each R.sup.2 and R.sup.3 are independently selected from hydrogen,
(C.sub.1-C.sub.7)alkyl, --O--(C.sub.1-C.sub.7)alkyl,
--(C.sub.0-C.sub.6) alkylene-carbocyclyl,
--(C.sub.0-C.sub.6)alkylene-heterocyclyl,
--(C.sub.1-C.sub.6)alkylene-O-carbocyclyl,
--(C.sub.1-C.sub.6)alkylene-O-heterocyclyl,
--S(O).sub.m--(C.sub.1-C.sub.6)alkyl,
--(C.sub.0-C.sub.4)alkylene-S(O).sub.m-carbocyclyl, and
--(C.sub.0-C.sub.4)alkylene-S(O).sub.m-heterocyclyl; or R.sup.2 and
R.sup.3, taken together with the nitrogen atom to which they are
bound form a heterocyclyl, wherein the heterocyclyl optionally
comprises 1 to 4 additional heteroatoms independently selected from
N, S and O; each R.sup.4 is independently selected from hydrogen,
(C.sub.1-C.sub.6)alkyl, --O--(C.sub.1-C.sub.6)alkyl,
--(C.sub.0-C.sub.6) alkylene-carbocyclyl, and
--(C.sub.0-C.sub.6)alkylene-heterocyclyl; each R.sup.5a and each
R.sup.5b is independently selected from hydrogen,
(C.sub.1-C.sub.6)alkyl, carbocyclyl, heterocyclyl or a naturally
occurring amino acid side chain moiety, or R.sup.5a and R.sup.5b
taken together with the carbon atom to which they are bound form a
3-7 membered non-aromatic carbocyclyl or a 4-7 membered
non-aromatic heterocyclyl, wherein the heterocyclyl formed by
R.sup.5a and R.sup.5b optionally comprises one to two additional
heteroatoms independently selected from N, S and O; R.sup.F is
selected from hydrogen, (C.sub.1-C.sub.7)alkyl, carbocyclyl or
heteroaryl, wherein: each carbocyclyl or heterocyclyl is optionally
and independently substituted with one or more substituents
independently selected from halo, --(C.sub.1-C.sub.4)alkyl, --OH,
.dbd.O, --O--(C.sub.1-C.sub.4)alkyl,
--(C.sub.1-C.sub.4)alkylene-O--(C.sub.1-C.sub.4)alkyl,
halo-substituted-(C.sub.1-C.sub.4)alkyl,
halo-substituted-O--(C.sub.1-C.sub.4)alkyl,
--C(O)--(C.sub.1-C.sub.4)alkyl,
--C(O)-(fluoro-substituted-(C.sub.1-C.sub.4)alkyl),
--S(O).sub.m--(C.sub.1-C.sub.4)alkyl, --N(R.sup.G)(R.sup.G) and CN;
the heterocyclyl in --N(R.sup.F)--C(O)-heterocyclyl represented by
Y is optionally substituted with carbocyclyl or heterocyclyl in
addition to the substituents described above, wherein the
carbocyclyl or heterocyclyl is optionally substituted with one or
more substituentts independently selected from halo,
(C.sub.1-C.sub.4)alkyl, --OH, .dbd.O, --O--(C.sub.1-C.sub.4)alkyl,
--(C.sub.1-C.sub.4)alkylene-O--(C.sub.1-C.sub.4)alkyl,
halo-substituted-(C.sub.1-C.sub.4)alkyl,
halo-substituted-O--(C.sub.1-C.sub.4)alkyl,
--C(O)--(C.sub.1-C.sub.4)alkyl,
--C(O)-(fluoro-substituted-(C.sub.1-C.sub.4)alkyl),
--S(O).sub.m--(C.sub.1-C.sub.4)alkyl, --N(R.sup.G)(R.sup.G) and CN;
each alkyl is optionally and independently substituted with one or
more substituents independently selected from halo,
--(C.sub.1-C.sub.4)alkyl, --OH, --O--(C.sub.1-C.sub.4)alkyl,
--(C.sub.1-C.sub.4)alkylene-O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl,
--S(O).sub.m--(C.sub.1-C.sub.4)alkyl and --N(R.sup.G)(R.sup.G);
each R.sup.G is hydrogen or (C.sub.1-C.sub.4)alkyl, wherein each
alkyl in the group represented by R.sup.G is optionally and
independently substituted with one or more substituents
independently selected from --(C.sub.1-C.sub.4)alkyl,
(C.sub.3-C.sub.6)cycloalkyl, halo, --OH,
--O--(C.sub.1-C.sub.4)alkyl, and
--(C.sub.1-C.sub.4)alkylene-O--(C.sub.1-C.sub.4)alkyl; and each m
is 1 or 2.
2. The compound of claim 1, wherein: Y is selected from hydrogen,
--(C.sub.1-C.sub.4)alkylene-N(R.sup.2)(R.sup.3),
--CH.dbd.N--OR.sup.2, --C(O)--N(R.sup.2)(R.sup.4), --NO.sub.2,
--COOH, --OH, --N.dbd.CH--N(R.sup.2)(R.sup.3),
--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--[C(R.sup.5a)(R.sup.5b)].sub.1-4--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--(C.sub.1-C.sub.6)alkyl,
--N(R.sup.F)--C(O)-heterocyclyl, --N(R.sup.F)--C(O)-carbocyclyl,
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4)alkylene-N(R.sup.2)(R.sup.3),
--N(R.sup.F)--S(O).sub.m--N(R.sup.2)(R.sup.4), and
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4) alkylene-carbocyclyl,
wherein: each R.sup.2 is independently selected from hydrogen, and
(C.sub.1-C.sub.3)alkyl; each R.sup.3 is independently selected from
hydrogen, (C.sub.1-C.sub.6)alkyl, --O--(C.sub.1-C.sub.6)alkyl,
--(C.sub.0-C.sub.6) alkylene-carbocyclyl,
--(C.sub.0-C.sub.6)alkylene-heterocyclyl,
--S(O).sub.m---(C.sub.1-C.sub.6)alkyl, --S(O).sub.m-carbocyclyl,
and --S(O).sub.m-heterocyclyl; and each R.sup.4 is independently
selected from hydrogen, (C.sub.1-C.sub.6)alkyl,
--O--(C.sub.1-C.sub.6)alkyl, --(C.sub.0-C.sub.6)
alkylene-carbocyclyl, and --(C.sub.0-C.sub.6)alkylene-heterocyclyl;
or R.sup.2 and R.sup.3, taken together with the nitrogen atom to
which they are bound form a heterocyclyl, wherein the heterocylyl
optionally comprises 1 to 4 additional heteroatoms independently
selected from N, S and O; each R.sup.5a and each R.sup.5b is
independently selected from hydrogen, (C.sub.1-C.sub.6)alkyl,
carbocyclyl, heterocyclyl, or a naturally occurring amino acid side
chain moiety, or R.sup.5a and R.sup.5b taken together with the
carbon atom to which they are bound form a 3-7 membered saturated
carbocyclyl or a 4-7 membered saturated heterocyclyl, wherein the
saturated heterocyclyl formed by R.sup.5a and R.sup.5b optionally
comprises one to two additional heteroatoms independently selected
from N, S and O, R.sup.F is hydrogen or (C.sub.1-C.sub.3)alkyl; and
each m is 1 or 2, wherein: each carbocyclyl or heterocyclyl is
optionally and independently substituted with one or more
substituents independently selected from chloro, fluoro,
(C.sub.1-C.sub.4)alkyl, --OH, --O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl,
--C(O)--(C.sub.1-C.sub.4)alkyl,
--C(O)-(fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.G)(R.sup.G); and each alkyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, chloro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
3. The compound of claim 2, wherein: Y is selected from hydrogen,
--N(R.sup.2)(R.sup.3),
--NH--C(O)--(CH.sub.2).sub.1-4--N(R.sup.2)(R.sup.3),
--NH--C(O)-heterocyclyl, --NH--C(O)-carbocyclyl, and
--NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl, wherein: each R.sup.2 is
independently selected from hydrogen, and (C.sub.1-C.sub.3)alkyl;
and each R.sup.3 is independently selected from hydrogen,
(C.sub.1-C.sub.6)alkyl, and
--(C.sub.0-C.sub.6)alkylene-carbocyclyl; or R.sup.2 and R.sup.3
taken together with the nitrogen atom to which they are bound form
a heterocyclyl, wherein the heterocylyl optionally comprises 1 to 4
additional heteroatoms independently selected from N, S and O;
wherein each carbocyclyl or heterocyclyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, (C.sub.1-C.sub.4)alkyl,
--O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.G)(R.sup.G); and each alkyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, --O--(C.sub.1-C.sub.4)alkyl,
and fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
4. The compound of claim 3, wherein: Y is selected from hydrogen,
--NH.sub.2, --NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3), ##STR00281##
--NH--C(O)-phenyl, --NH--C(O)-thienyl and
--NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl, wherein: each R.sup.2 is
independently selected from hydrogen, and (C.sub.1-C.sub.2)alkyl;
and each R.sup.3 is independently selected from hydrogen,
(C.sub.1-C.sub.6)alkyl, and --(C.sub.0-C.sub.1)alkylene-cycloalkyl;
or R.sup.2 and R.sup.3 taken together with the nitrogen atom to
which they are bound form a saturated heterocyclyl; ring A
represents a 4-7 membered saturated heterocyclyl; R.sup.3' is
hydrogen or methyl; each cycloalkyl, phenyl or heterocyclyl is
optionally and independently substituted with one or more
substituents independently selected from fluoro,
(C.sub.1-C.sub.4)alkyl, --O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.2)(R.sup.2); and each alkyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, --O--(C.sub.1-C.sub.4)alkyl,
and fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
5. The compound of claim 2, wherein: X is --OCH.sub.3; Y is
selected from --NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3),
##STR00282## and --NH--S(O).sub.2---(C.sub.1-C.sub.6)alkyl, wherein
the alkyl group in --NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl is
optionally substituted with fluoro; each R.sup.2 is independently
selected from hydrogen and --CH.sub.3; each R.sup.3 is
independently selected from (C.sub.1-C.sub.6)alkyl, and
--(C.sub.0-C.sub.1)alkylene-carbocyclyl; or R.sup.2 and R.sup.3,
taken together with the nitrogen atom to which they are bound form
a saturated heterocyclyl optionally substituted with fluoro; ring A
represents a 4-7 membered saturated heterocyclyl; R.sup.3' is
hydrogen or methyl; each carbocyclyl or heterocyclyl is optionally
and independently substituted with one or more substituents
independently selected from fluoro, (C.sub.1-C.sub.4)alkyl,
--O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.2)(R.sup.2); and each alkyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, --O--(C.sub.1-C.sub.4)alkyl,
and fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
6. The compound of claim 2, wherein: X is --OCF.sub.3; Y is
selected from --NH.sub.2,
--NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3), ##STR00283##
--NH--C(O)-phenyl, and --NH--C(O)-thienyl, wherein: the phenyl in
the group represented by Y is optionally substituted with
--OCH.sub.3 or --N(CH.sub.3).sub.2; each R.sup.2 is independently
selected from hydrogen and --CH.sub.3; each R.sup.3 is
independently selected from (C.sub.1-C.sub.6)alkyl and
--(C.sub.0-C.sub.1)alkylene-carbocyclyl, wherein the
(C.sub.1-C.sub.6)alkyl is optionally substituted with fluoro or
--OCH.sub.3; or R.sup.2 and R.sup.3, taken together with the
nitrogen atom to which they are bound form a saturated heterocyclyl
optionally substituted with fluoro or --OCH.sub.3; ring A
represents a 4-7 membered saturated heterocyclyl; R.sup.3' is
hydrogen or methyl; each carbocyclyl or heterocyclyl is optionally
and independently substituted with one or more substituents
independently selected from fluoro, (C.sub.1-C.sub.4)alkyl,
--O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.G)(R.sup.G); and each alkyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, --O--(C.sub.1-C.sub.4)alkyl,
and fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
7. (canceled)
8. The compound of claim 2, wherein the compound is selected from
any one of compound numbers 101, 102, 103, 104, 105, 106, 107, 109,
112, 114, 115, 120, 123, 125, 126, 127, 128, 130, 131, 132, 133,
135, 136, 142, 144, 145, 146, 147, 148, 152, 154, 169, 170, 173,
209, 210, 212, 213, 214, 217, 218, 220, 221, 222, 224, 226, 230,
234, 239, 243, 244, 245, 246, 247, 248, 250, 251, 252, 257, 258,
259, 260, 261, and 263, or a pharmaceutically acceptable salt
thereof.
9. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier or diluent and a compound of claim 1.
10. A method for treating or preventing an infection or
colonization in a subject comprising administering to the subject
an effective amount of a compound of claim 1.
11. The method of claim 10, wherein the infection is caused by a
Gram-positive organism.
12. The method of claim 11, wherein the Gram-positive organism is
selected from the group consisting of Staphylococcus spp.,
Streptococcus spp., Propionibacterium spp., Enterococcus spp.,
Bacillus spp., Corynebacterium spp., Nocardia spp., Clostridium
spp., Actinobacteria spp., and Listeria spp.
13. The method of claim 10, wherein the infection is caused by a
Gram-negative organism.
14. The method of claim 13, wherein the Gram-negative organism is
selected form the group consisting of Enterobactericeae,
Bacteroidaceae, Vibrionaceae, Pasteurellae, Pseudomonadaceae,
Neisseriaceae, Rickettsiae, Moraxellaceae any species of Proteeae,
Acinetobacter spp., Helicobacter spp., and Campylobacter spp.
15. The method of claim 10, wherein the infection is caused by an
organism selected from the group consisting of rickettsiae,
chlamydiae, Legionella spp. Mycoplasma spp., and any other
intracellular pathogens.
16-22. (canceled)
Description
RELATED APPLICATION
[0001] This application is a continuation of U.S. application Ser.
No. 13/391,407, filed May 2, 2012, which is the U.S. National Stage
of International Application No. PCT/US2010/047035, filed Aug. 27,
2010, which designates the U.S., published in English, which claims
the benefit of U.S. Provisional Application No. 61/275,507, filed
on Aug. 28, 2009. The entire teachings of the above application are
incorporated herein by reference.
BACKGROUND OF THE INVENTION
[0002] The tetracyclines are broad spectrum anti microbial agents
that are widely used in human and veterinary medicine. The total
production of tetracyclines by fermentation or semi synthesis is
measured in the thousands of metric tons per year.
[0003] The widespread use of tetracyclines for therapeutic purposes
has led to the emergence of resistance to these antibiotics, even
among highly susceptible bacterial species. Therefore, there is
need for new tetracycline analogs with improved antibacterial
activities and efficacies against other tetracycline responsive
diseases or disorders.
SUMMARY OF THE INVENTION
[0004] One embodiment of the present invention is directed to a
compound represented by Structural Formula (I):
##STR00002##
[0005] or a pharmaceutically acceptable salt thereof.
[0006] X is selected from --CF.sub.3, --CN, --OCF.sub.3 and
--OCH.sub.3.
[0007] Y is selected from hydrogen, --(C.sub.1-C.sub.7)alkyl,
carbocyclyl, --(C.sub.1-C.sub.4)alkylene-N(R.sup.2)(R.sup.3),
--(C.sub.1-C.sub.4)alkylene-N(R.sup.F)--C(O)--[C(R.sup.5a)(R.sup.5b)].sub-
.0-4 --N(R.sup.2)(R.sup.3), --CH.dbd.N--OR.sup.2,
--C(O)--N(R.sup.2)(R.sup.4), --NO.sub.2, --COOH, --OH,
--N.dbd.CH--N(R.sup.2)(R.sup.3), --N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--[C(R.sup.5a)(R.sup.5b)].sub.1-4--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--(C.sub.1-C.sub.6)alkyl,
--N(R.sup.F)--C(O)-heterocyclyl, --N(R.sup.F)--C(O)-carbocyclyl,
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4)
alkylene-N(R.sup.2)(R.sup.3),
--N(R.sup.F)--S(O).sub.m--N(R.sup.2)(R.sup.4), and
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4)alkylene-carbocyclyl.
[0008] Each R.sup.2 and R.sup.3 are independently selected from
hydrogen, (C.sub.1-C.sub.7)alkyl, --O--(C.sub.1-C.sub.7)alkyl,
--(C.sub.0-C.sub.6) alkylene-carbocyclyl,
--(C.sub.0-C.sub.6)alkylene-heterocyclyl,
--(C.sub.1-C.sub.6)alkylene-O-carbocyclyl,
--(C.sub.1-C.sub.6)alkylene-O-heterocyclyl,
--S(O).sub.m---(C.sub.1-C.sub.6)alkyl,
--(C.sub.0-C.sub.4)alkylene-S(O).sub.m-carbocyclyl, and
--(C.sub.0-C.sub.4)alkylene-S(O).sub.m-heterocyclyl.
[0009] Each R.sup.4 is independently selected from hydrogen,
(C.sub.1-C.sub.6)alkyl, --O--(C.sub.1-C.sub.6)alkyl,
--(C.sub.0-C.sub.6) alkylene-carbocyclyl and
--(C.sub.0-C.sub.6)alkylene-heterocyclyl.
[0010] Each R.sup.5a and each R.sup.5b is independently selected
from hydrogen, (C.sub.1-C.sub.6)alkyl, carbocyclyl, heterocyclyl,
or a naturally occurring amino acid side chain moiety, or
[0011] R.sup.5a and R.sup.5b taken together with the carbon atom to
which they are bound form a 3-7 membered non-aromatic carbocyclyl
or a 4-7 membered non-aromatic heterocyclyl, wherein the
heterocyclyl formed by R.sup.5a and R.sup.5b optionally comprises
one to two additional heteroatoms independently selected from N, S
and O.
[0012] R.sup.F is selected from hydrogen, (C1 C7)alkyl, carbocyclyl
or heteroaryl
[0013] Alternatively, R.sup.2 and R.sup.3, taken together with the
nitrogen atom to which they are bound form a heterocyclyl, wherein
the heterocyclyl optionally comprises 1 to 4 additional heteroatoms
independently selected from N, S and O.
[0014] Each carbocyclyl or heterocyclyl described above (e.g., in
the groups represented by Y, R.sup.2, R.sup.3, R.sup.4, R.sup.5a,
R.sup.5b, --NR.sup.2R.sup.3 or R.sup.5a and R.sup.5b taken
together) is optionally and independently substituted with one or
more substituents independently selected from halo (e.g., chloro or
fluoro), --(C.sub.1-C.sub.4)alkyl, --OH, .dbd.O,
--O--(C.sub.1-C.sub.4)alkyl,
--(C.sub.1-C.sub.4)alkylene-O--(C.sub.1-C.sub.4)alkyl,
halo-substituted-(C.sub.1-C.sub.4)alkyl,
halo-substituted-O-(C.sub.1-C.sub.4)alkyl,
--C(O)--(C.sub.1-C.sub.4)alkyl,
--C(O)-(fluoro-substituted-(C.sub.1-C.sub.4)alkyl),
--S(O).sub.m--(C.sub.1-C.sub.4)alkyl, --N(R.sup.G)(R.sup.G) and
CN.
[0015] The heterocyclyl in --N(R.sup.F)--C(O)-heterocyclyl
represented by Y is optionally substituted with carbocyclyl or
heterocyclyl in addition to the substituents described above,
wherein the carbocyclyl or heterocyclyl is optionally substituted
with one or more substituentts independently selected from halo
(e.g., chloro or fluoro), (C.sub.1-C.sub.4)alkyl, --OH, .dbd.O,
--O--(C.sub.1-C.sub.4)alkyl,
--(C.sub.1-C.sub.4)alkylene-O--(C.sub.1-C.sub.4)alkyl,
halo-substituted-(C.sub.1-C.sub.4)alkyl,
halo-substituted-O-(C.sub.1-C.sub.4)alkyl,
--C(O)--(C.sub.1-C.sub.4)alkyl,
--C(O)-(fluoro-substituted-(C.sub.1-C.sub.4)alkyl),
--S(O).sub.m--(C.sub.1-C.sub.4)alkyl, --N(R.sup.G)(R.sup.G) and
CN.
[0016] Each alkyl described above (e.g., in the groups represented
by Y, R.sup.2, R.sup.3, R.sup.4, R.sup.5a or R.sup.5b) is
optionally and independently substituted with one or more
substituents independently selected from halo (e.g., fluoro or
chloro), --(C.sub.1-C.sub.4)alkyl, --OH,
--O--(C.sub.1-C.sub.4)alkyl,
--(C.sub.1-C.sub.4)alkylene-O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl,
--S(O).sub.m--(C.sub.1-C.sub.4)alkyl and -N(RG)(RG).
[0017] Each R.sup.G is hydrogen or (C.sub.1-C.sub.4)alkyl, wherein
each alkyl in the group represented by R.sup.G is optionally and
independently substituted with one or more substituents
independently selected from --(C.sub.1-C.sub.4)alkyl,
(C.sub.3-C.sub.6)cycloalkyl, halo, --OH,
--O--(C.sub.1-C.sub.4)alkyl, and
--(C.sub.1-C.sub.4)alkylene-O--(C.sub.1-C.sub.4)alkyl.
[0018] Each m is 1 or 2.
[0019] Another embodiment of the present invention is directed to a
pharmaceutical composition comprising a pharmaceutically acceptable
carrier or diluent and a compound represented by Structural Formula
(I) or a pharmaceutically acceptable salt thereof. The
pharmaceutical composition is used in therapy, such as treating an
infection (e.g., a bacterial infection) in a subject.
[0020] Another embodiment of the present invention is a method of
treating an infection (e.g., a bacterial infection) in a subject
comprising administering to the subject an effective amount of a
compound represented by Structural Formula (I) or a
pharmaceutically acceptable salt thereof.
[0021] Another embodiment of the present invention is a method of
preventing an infection (e.g., a bacterial infection) in a subject
comprising administering to the subject an effective amount of a
compound represented by Structural Formula (I) or a
pharmaceutically acceptable salt thereof.
[0022] Another embodiment of the present invention is the use of a
compound represented by Structural Formula (I) or a
pharmaceutically acceptable salt thereof for the manufacture of a
medicament for treating an infection (e.g., a bacterial infection)
in a subject.
[0023] Another embodiment of the present invention is the use of a
compound represented by Structural Formula (I) or a
pharmaceutically acceptable salt thereof for the manufacture of a
medicament for preventing an infection (e.g., a bacterial
infection) in a subject.
[0024] Another embodiment of the present invention is the use of a
compound represented by Structural Formula (I) or a
pharmaceutically acceptable salt thereof in therapy, such as
treating or preventing an infection (e.g., a bacterial infection)
in a subject.
DETAILED DESCRIPTION OF THE INVENTION
[0025] The present invention is directed to a compound represented
by Structural Formula (I) or a pharmaceutically acceptable salt
thereof. Values and alternative values for the variables in
Structural Formula (I) and for each of the embodiments described
herein are defined as the following:
[0026] X is selected from --CF.sub.3, --CN, --OCF.sub.3 and
--OCH.sub.3. Alternatively, X is --CF.sub.3. In another
alternative, X is --CN. In another alternative, X is --OCF.sub.3.
In another alternative, X is --OCH.sub.3.
[0027] Y is selected from hydrogen, --(C.sub.1-C.sub.7)alkyl,
carbocyclyl, --(C.sub.1-C.sub.4)alkylene-N(R.sup.2)(R.sup.3),
--(C.sub.1-C.sub.4)alkylene-N(R.sup.F)--C(O)--[C(R.sup.5a)(R.sup.5b)].sub-
.0-4--N(R.sup.2)(R.sup.3), --CH.dbd.N--OR.sup.2,
--C(O)--N(R.sup.2)(R.sup.4), --NO.sub.2, --COOH, --OH,
--N.dbd.CH--N(R.sup.2)(R.sup.3), --N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--[C(R.sup.5a)(R.sup.5b)].sub.1-4--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--(C.sub.1-C.sub.6)alkyl,
--N(R.sup.F)--C(O)-heterocyclyl, --N(R.sup.F)--C(O)-carbocyclyl,
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4)alkylene-N(R.sup.2)(R.sup.3),
--N(R.sup.F)--S(O).sub.m--N(R.sup.2)(R.sup.4), and
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4)alkylene-carbocyclyl. In
one embodiment, Y is selected from hydrogen,
--(C.sub.1-C.sub.4)alkylene-N(R.sup.2)(R.sup.3),
--CH.dbd.N--OR.sup.2, --C(O)--N(R.sup.2)(R.sup.4), --NO.sub.2,
--COOH, --OH, --N.dbd.CH--N(R.sup.2)(R.sup.3),
--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--[C(R.sup.5a)(R.sup.5b)].sub.1-4--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--(C.sub.1-C.sub.6)alkyl,
--N(R.sup.F)--C(O)-heterocyclyl, --N(R.sup.F)--C(O)-carbocyclyl,
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4)
alkylene-N(R.sup.2)(R.sup.3),
--N(R.sup.F)--S(O).sub.m--N(R.sup.2)(R.sup.4), and
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4)alkylene-carbocyclyl,
wherein R.sup.F in the groups represented by Y is hydrogen or
(C.sub.1-C.sub.3)alkyl. Alternatively, Y is selected from hydrogen,
--N(R.sup.2)(R.sup.3),
--NH--C(O)--(CH.sub.2).sub.1-4--N(R.sup.2)(R.sup.3),
--NH--C(O)-heterocyclyl, --NH--C(O)-carbocyclyl, and
--NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl. In another alternative, Y
is selected from hydrogen, --NH.sub.2,
--NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3),
##STR00003##
--NH--C(O)-phenyl, --NH--C(O)-thienyl and
--NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl, wherein ring A represents
a 4-7 membered saturated heterocyclyl and R.sup.3' is is hydrogen,
(C.sub.1-C.sub.6)alkyl, carbocyclyl or heterocyclyl, wherein the
alkyl, carbocyclyl or heterocyclyl are optionally and independently
substituted with one or more substituents independently selected
from the group described above for Structural Formula (I). In
another alternative, Y is selected from
--NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3),
##STR00004##
and --NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl, wherein ring A and
R.sup.3' are as described above; and the alkyl group in
--NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl is optionally substituted
with fluoro. In another alternative, Y is selected from --NH.sub.2,
--NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3),
##STR00005##
--NH--C(O)-phenyl and --NH--C(O)-thienyl, wherein ring A and
R.sup.3' are as described above; and the phenyl in the
--NH--C(O)-phenyl group is optionally substituted with --OCH.sub.3
or --N(CH.sub.3).sub.2. In another alternative, Y is selected from
--NH.sub.2, --NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3), and
##STR00006##
wherein ring A and R.sup.3' are as described above.
[0028] Each R.sup.2 and R.sup.3 are independently selected from
hydrogen, (C.sub.1-C.sub.7)alkyl, --O--(C.sub.1-C.sub.7)alkyl,
--(C.sub.0-C.sub.6) alkylene-carbocyclyl,
--(C.sub.0-C.sub.6)alkylene-heterocyclyl,
--(C.sub.1-C.sub.6)alkylene-O-carbocyclyl,
--(C.sub.1-C.sub.6)alkylene-O-heterocyclyl,
--S(O).sub.m---(C.sub.1-C.sub.6)alkyl,
--(C.sub.0-C.sub.4)alkylene-S(O).sub.m-carbocyclyl, and
--(C.sub.0-C.sub.4)alkylene-S(O).sub.m-heterocyclyl.
[0029] In one embodiment, each R.sup.2 is independently selected
from hydrogen, and (C.sub.1-C.sub.3)alkyl; and each R.sup.3 is
independently selected from hydrogen, (C.sub.1-C.sub.6)alkyl,
--O--(C.sub.1-C.sub.6)alkyl, --(C.sub.0-C.sub.6)
alkylene-carbocyclyl, --(C.sub.0-C.sub.6)alkylene-heterocyclyl,
--S(O).sub.m--(C.sub.1-C.sub.6)alkyl, --S(O).sub.m-carbocyclyl and
--S(O).sub.m-heterocyclyl; or R.sup.2 and R.sup.3, taken together
with the nitrogen atom to which they are bound form a heterocyclyl,
wherein the heterocyclyl optionally comprises 1 to 4 additional
heteroatoms independently selected from N, S and O.
[0030] Alternatively, each R.sup.2 is independently selected from
hydrogen and (C.sub.1-C.sub.3)alkyl; and each R.sup.3 is
independently selected from hydrogen, (C.sub.1-C.sub.6)alkyl, and
--(C.sub.0-C.sub.6)alkylene-carbocyclyl; or R.sup.2 and R.sup.3
taken together with the nitrogen atom to which they are bound form
a heterocyclyl, wherein the heterocylyl optionally comprises 1 to 4
additional heteroatoms independently selected from N, S and O.
[0031] In another alternative, each R.sup.2 is independently
selected from hydrogen, and (C.sub.1-C.sub.2)alkyl; and each
R.sup.3 is independently selected from hydrogen,
(C.sub.1-C.sub.6)alkyl, and --(C.sub.0-C.sub.1)
alkylene-cycloalkyl; or R.sup.2 and R.sup.3 taken together with the
nitrogen atom to which they are bound form a saturated
heterocyclyl.
[0032] In another alternative, each R.sup.2 is independently
selected from hydrogen and --CH.sub.3; and each R.sup.3 is
independently selected from (C.sub.1-C.sub.6)alkyl, and
--(C.sub.0-C.sub.1)alkylene-carbocyclyl; or R.sup.2 and R.sup.3,
taken together with the nitrogen atom to which they are bound form
a saturated heterocyclyl optionally substituted with fluoro. In
another alternative, each R.sup.2 is independently selected from
hydrogen and --CH.sub.3; and each R.sup.3 is independently selected
from (C.sub.1-C.sub.6)alkyl optionally substituted with fluoro or
--OCH.sub.3, and --(C.sub.0-C.sub.1)alkylene-carbocyclyl; or
R.sup.2 and R.sup.3, taken together with the nitrogen atom to which
they are bound form a saturated heterocyclyl optionally substituted
with fluoro or --OCH.sub.3.
[0033] Each R.sup.4 is independently selected from hydrogen,
(C.sub.1-C.sub.6)alkyl, --O--(C.sub.1-C.sub.6)alkyl,
--(C.sub.0-C.sub.6) alkylene-carbocyclyl, and
--(C.sub.0-C.sub.6)alkylene-heterocyclyl.
[0034] Each R.sup.5a and each R.sup.5b is independently selected
from hydrogen, (C.sub.1-C.sub.6)alkyl, carbocyclyl, heterocyclyl or
a naturally occurring amino acid side chain moiety, or R.sup.5a and
R.sup.5b taken together with the carbon atom to which they are
bound form a 3-7 membered non-aromatic carbocyclyl or a 4-7
membered non-aromatic heterocyclyl, wherein the heterocyclyl formed
by R.sup.5a and R.sup.5b optionally comprises one to two additional
heteroatoms independently selected from N, S and O. In one
embodiment, R.sup.5a and R.sup.5b taken together with the carbon
atom to which they are bound form a 3-7 membered saturated
carbocyclyl or a 4-7 membered saturated heterocyclyl, wherein the
heterocyclyl formed by R.sup.5a and R.sup.5b optionally comprises
one to two additional heteroatoms independently selected from N, S
and O. In one embodiment, each R.sup.5a and each R.sup.5b is
independently hydrogen, (C.sub.1-C.sub.6)alkyl or a naturally
occurring amino acid side chain moiety. In another embodiment,
R.sup.5a and R.sup.5b are both hydrogen.
[0035] Each carbocyclyl or heterocyclyl described above (e.g., in
the groups represented by Y, R.sup.3, R.sup.4, R.sup.5a, R.sup.5b,
--NR.sup.2R.sup.3 or R.sup.5a and R.sup.5b taken together) is
optionally and independently substituted with one or more
substituents independently selected from halo (e.g., chloro or
fluoro), --(C.sub.1-C.sub.4)alkyl, --OH, .dbd.O,
--O--(C.sub.1-C.sub.4)alkyl,
--(C.sub.1-C.sub.4)alkylene-O--(C.sub.1-C.sub.4)alkyl,
halo-substituted-(C.sub.1-C.sub.4)alkyl,
halo-substituted-O--(C.sub.1-C.sub.4)alkyl,
--C(O)--(C.sub.1-C.sub.4)alkyl,
--C(O)-(fluoro-substituted-(C.sub.1-C.sub.4)alkyl),
--S(O).sub.m--(C.sub.1-C.sub.4)alkyl, --N(R.sup.G)(R.sup.G) and CN.
The heterocyclyl in --N(R.sup.F)--C(O)-heterocyclyl represented by
Y is optionally substituted with carbocyclyl or heterocyclyl in
addition to the substituents described above, wherein the
carbocyclyl or heterocyclyl is optionally substituted with one or
more substituents independently selected from chloro, fluoro,
(C.sub.1-C.sub.4)alkyl, --OH, --O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl,
--C(O)--(C.sub.1-C.sub.4)alkyl,
--C(O)-(fluoro-substituted-(C.sub.1-C.sub.4)alkyl), and
--N(R.sup.G)(R.sup.G).
[0036] In one embodiment, each carbocyclyl, or heterocyclyl
described above is optionally and independently substituted with
one or more substituents independently selected from chloro,
fluoro, (C.sub.1-C.sub.4)alkyl, --OH, --O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl,
--C(O)--(C.sub.1-C.sub.4)alkyl,
--C(O)-(fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.2)(R.sup.2).
[0037] Each alkyl described above (e.g., in the groups represented
by Y, R.sup.2, R.sup.3, R.sup.4, R.sup.5a or R.sup.5b) is
optionally and independently substituted with one or more
substituents independently selected from halo (e.g., fluoro or
chloro), --(C.sub.1-C.sub.4)alkyl, --OH,
--O--(C.sub.1-C.sub.4)alkyl,
--(C.sub.1-C.sub.4)alkylene-O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl,
--S(O).sub.m--(C.sub.1-C.sub.4)alkyl and --N(R.sup.G)(R.sup.G).
[0038] In one embodiment, each alkyl described above is optionally
and independently substituted with one or more substituents
independently selected from fluoro, chloro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
[0039] Alternatively, each carbocyclyl (e.g., cycloalkyl or
phenyl), or heterocyclyl descrubed above is optionally and
independently substituted with one or more substituents
independently selected from fluoro, (C.sub.1-C.sub.4)alkyl,
--O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.G)(R.sup.G); and each alkyl described above is optionally
and independently substituted with one or more substituents
independently selected from fluoro, --O--(C.sub.1-C.sub.4)alkyl,
and fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
[0040] Alternatively, each carbocyclyl described above is
(C.sub.3-C.sub.6)cycloalkyl or phenyl; each heterocyclyl described
above is independently selected from azetidinyl, morphinyl,
piperazinyl, piperidinyl, pyrrolidinyl, azepanyl and
octahydrocyclopenta[c]pyrrolyl and thienyl; each of said
carbocyclyl, cycloalkyl, heterocyclyl and phenyl is optionally
substituted as described in the previous paragraph; and each said
alkyl is optionally substituted as described in the previous
paragraph.
[0041] Each R.sup.G is hydrogen or (C.sub.1-C.sub.4)alkyl, wherein
each alkyl in the group represented by R.sup.G is optionally and
independently substituted with one ore more substituents
independently selected from --(C.sub.1-C.sub.4)alkyl,
(C.sub.3-C.sub.6)cycloalkyl, halo, --OH,
--O--(C.sub.1-C.sub.4)alkyl, and
--(C.sub.1-C.sub.4)alkylene-O--(C.sub.1-C.sub.4)alkyl.
[0042] Each m is 1 or 2. In one embodiment, m is 2.
[0043] In a first alternative embodiment, the compound of the
present invention is represented by Structural Formula (I), or a
pharmaceutically acceptable salt thereof, wherein:
[0044] X is selected from --CF.sub.3, --CN, --OCF.sub.3 and
--OCH.sub.3; and
[0045] Y is selected from hydrogen,
--(C.sub.1-C.sub.4)alkylene-N(R.sup.2)(R.sup.3),
--CH.dbd.N--OR.sup.2, --C(O)--N(R.sup.2)(R.sup.4), --NO.sub.2,
--COOH, --OH, --N.dbd.CH--N(R.sup.2)(R.sup.3),
--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--[C(R.sup.5a)(R.sup.5b)].sub.1-4--N(R.sup.2)(R.sup.3),
--N(R.sup.F)--C(O)--(C.sub.1-C.sub.6)alkyl,
--N(R.sup.F)--C(O)-heterocyclyl, --N(R.sup.F)--C(O)-carbocyclyl,
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4)alkylene-N(R.sup.2)(R.sup.3),
--N(R.sup.F)--S(O).sub.m--N(R.sup.2)(R.sup.4), and
--N(R.sup.F)--S(O).sub.m--(C.sub.1-C.sub.4)alkylene-carbocyclyl,
wherein R.sup.F in the groups represented by Y is hydrogen or
(C.sub.1-C.sub.3)alkyl.
[0046] each R.sup.2 is independently selected from hydrogen, and
(C.sub.1-C.sub.3)alkyl;
[0047] each R.sup.3 is independently selected from hydrogen,
(C.sub.1-C.sub.6)alkyl, --O--(C.sub.1-C.sub.6)alkyl,
--(C.sub.0-C.sub.6) alkylene-carbocyclyl,
--(C.sub.0-C.sub.6)alkylene-heterocyclyl,
--S(O).sub.m--(C.sub.1-C.sub.6)alkyl, --S(O).sub.m-carbocyclyl, and
--S(O).sub.m-heterocyclyl; or R.sup.2 and R.sup.3, taken together
with the nitrogen atom to which they are bound form a heterocyclyl,
wherein the heterocylyl optionally comprises 1 to 4 additional
heteroatoms independently selected from N, S and O.
[0048] each R.sup.4 is independently selected from hydrogen,
(C.sub.1-C.sub.6)alkyl, --O--(C.sub.1-C.sub.6)alkyl,
--(C.sub.0-C.sub.6) alkylene-carbocyclyl, and
--(C.sub.0-C.sub.6)alkylene-heterocyclyl; or
[0049] each R.sup.5a and each R.sup.5b is independently selected
from hydrogen, (C.sub.1-C.sub.6)alkyl, carbocyclyl or heterocyclyl
or a naturally occurring amino acid side chain moiety, or
[0050] R.sup.5a and R.sup.5b taken together with the carbon atom to
which they are bound form a 3-7 membered non-aromatic carbocyclyl
or a 4-7 membered non-aromatic heterocyclyl, wherein the
heterocyclyl formed by R.sup.5a and R.sup.5b optionally comprises
one to two additional heteroatoms independently selected from N, S
and O,
[0051] R.sup.F is hydrogen or (C.sub.1-C.sub.3)alkyl; and
[0052] each m is 1 or 2, wherein:
[0053] each carbocyclyl or heterocyclyl is optionally and
independently substituted with one or more substituents
independently selected from chloro, fluoro, (C.sub.1-C.sub.4)alkyl,
--OH, --O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl,
--C(O)--(C.sub.1-C.sub.4)alkyl,
--C(O)-(fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.G)(R.sup.G);
[0054] each alkyl is optionally and independently substituted with
one or more substituents independently selected from fluoro,
chloro, --O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl; and
[0055] each R.sup.G is hydrogen or (C.sub.1-C.sub.3)alkyl, wherein
each alkyl group represented by R.sup.G is optionally and
independently substituted with one or more substituents
independently selected from fluoro, chloro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
[0056] In a second alternative embodiment, the compound of the
present invention is represented by Structural Formula (I), or a
pharmaceutically acceptable salt thereof, wherein:
[0057] X is selected from --CF.sub.3, --CN, --OCF.sub.3 and
--OCH.sub.3.
[0058] Y is selected from hydrogen, --N(R.sup.2)(R.sup.3),
--NH--C(O)--(CH.sub.2).sub.1-4--N(R.sup.2)(R.sup.3),
--NH--C(O)-heterocyclyl, --NH--C(O)-carbocyclyl, and
--NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl, wherein:
[0059] each R.sup.2 is independently selected from hydrogen, and
(C.sub.1-C.sub.3)alkyl; and
[0060] each R.sup.3 is independently selected from hydrogen,
(C.sub.1-C.sub.6)alkyl, and
--(C.sub.0-C.sub.6)alkylene-carbocyclyl; or
[0061] R.sup.2 and R.sup.3 taken together with the nitrogen atom to
which they are bound form a heterocyclyl, wherein the heterocyclyl
optionally comprises 1 to 4 additional heteroatoms independently
selected from N, S and O;
[0062] each carbocyclyl or heterocyclyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, (C.sub.1-C.sub.4)alkyl,
--O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and -N(RG)(RG); and
[0063] each alkyl is optionally and independently substituted with
one or more substituents independently selected from fluoro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl. The remainder of the
variables are as described aobve in the first alternative
embodiment.
[0064] In a third alternative embodiment, the compound of the
present invention is represented by Structural Formula (I), or a
pharmaceutically acceptable salt thereof, wherein:
[0065] X is selected from --CF.sub.3, --CN, --OCF.sub.3 and
--OCH.sub.3.
[0066] Y is selected from hydrogen, --NH.sub.2,
--NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3),
##STR00007##
--NH--C(O)-phenyl, --NH--C(O)-thienyl and
--NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl, wherein:
[0067] each R.sup.2 is independently selected from hydrogen, and
(C.sub.1-C.sub.2)alkyl; and
[0068] each R.sup.3 is independently selected from hydrogen,
(C.sub.1-C.sub.6)alkyl, and --(C.sub.0-C.sub.1)alkylene-cycloalkyl;
or
[0069] R.sup.2 and R.sup.3 taken together with the nitrogen atom to
which they are bound form a saturated heterocyclyl;
[0070] ring A represents a 4-7 membered saturated heterocyclyl;
[0071] R.sup.3' is hydrogen or methyl;
[0072] each cycloalkyl, phenyl or heterocyclyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, (C.sub.1-C.sub.4)alkyl,
--O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.G)(R.sup.G); and
[0073] each alkyl is optionally and independently substituted with
one or more substituents independently selected from fluoro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl. The remainder of the
variables are as described aobve in the second alternative
embodiment.
[0074] In a fourth alternative embodiment, the compound of
Structural Formula (I) is represented by Structural Formula (II),
(III), (IV) or (V):
##STR00008##
[0075] or pharmaceutically acceptable salt thereof, wherein:
[0076] Y is selected from
--NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3), --NH--C(O)-heterocyclyl,
--NH--C(O)-phenyl, and
--NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl;
[0077] each R.sup.2 is independently hydrogen or a
(C.sub.1-C.sub.3)alkyl;
[0078] each R.sup.3 is independently a (C.sub.1-C.sub.6)alkyl or a
--(C.sub.0-C.sub.1)alkylene-carbocyclyl; or
[0079] R.sup.2 and R.sup.3, taken together with the nitrogen atom
to which they are bound form a saturated heterocyclyl. The
saturated heterocyclyl (e.g., azetidinyl, morphinyl, piperazinyl,
piperidinyl, pyrrolidinyl, azepanyl and
octahydrocyclopenta[c]pyrrolyl) is optionally substituted with one
or more substituents independently selected from fluoro,
(C.sub.1-C.sub.4)alkyl, --O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and --NN.sub.2,
--NH--(C.sub.1-C.sub.3)alkyl. The remainder of the variables are as
described above for Structural Formula (I) in the second
alternative embodiment.
[0080] In a fifth alternative embodiment, for compounds represented
by Structural Formulas (II)-(V), Y is:
##STR00009##
wherein ring A represents a 4-7 membered saturated heterocyclyl and
R.sup.3' is hydrogen, (C.sub.1-C.sub.6)alkyl, carbocyclyl or
heterocyclyl, wherein the alkyl, carbocyclyl or heterocyclyl are
optionally substituted with one or more substituents independently
selected from the groups described above for Structural Formula
(I). The remainder of the variables are as described above in the
fourth alternative embodiment. More specifically, R.sup.3' is
hydrogen or (C.sub.1-C.sub.6)alkyl. Even more specifically,
R.sup.3' is hydrogen or methyl. In another more specific
embodiment, ring A is selected from the group consisting of
azetidinyl, morphinyl, piperazinyl, piperidinyl, pyrrolidinyl,
azepanyl and octahydrocyclopenta[c]pyrrolyl, each of which is
optionally substituted with one or more substituents independently
selected from (C.sub.1-C.sub.3)alkyl, --F,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, such as --CF.sub.3,
--CH.sub.2CF.sub.3 or --CH.sub.2CHF.sub.2; and the remainder of the
variables are as described above for the fifth alternative
embodiment. Even more specifically, R.sup.3' is hydrogen or
(C.sub.1-C.sub.6)alkyl (e.g. methyl).
[0081] In a sixth alternative embodiment, for compounds represented
by Structural Formulas (II)-(V), Y is --NH--C(O)-heteroaryl, and
the remainder of the variables are as described in the third
alternative embodiment. More specifically, Y is
--NH--C(O)-thienyl.
[0082] In a seventh alternative embodiment, for compounds
represented by Structural Formulas (II)-(V), Y is
--NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3), R.sup.3 is a
(C.sub.1-C.sub.6)alkyl or a
--(C.sub.0-C.sub.1)alkylene-(C.sub.3-C.sub.6)cycloalkyl; and the
remainder of the variables are as described in the fourth
alternative embodiment.
[0083] In an eighth alternative embodiment, for compounds
represented by Structural Formulas (II)-(V):
[0084] Y is --NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3);
[0085] R.sup.2 is hydrogen or a (C.sub.1-C.sub.3)alkyl;
[0086] R.sup.3 is a (C.sub.1-C.sub.6)alkyl or a
--(C.sub.0-C.sub.1)alkylene-(C.sub.3-C.sub.6)cycloalkyl; or
[0087] R.sup.2 and R.sup.3, taken together with the nitrogen atom
to which they are bound form a saturated heterocyclyl optionally
substituted with one or more substituents independently selected
from the group consisting of (C.sub.1-C.sub.3)alkyl, --F,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, such as --CF.sub.3,
--CH.sub.2CF.sub.3 or --CH.sub.2CHF.sub.2.
[0088] Examples of the heterocyclyl include azetidinyl,
piperidinyl, pyrrolidinyl, azepanyl and
octahydrocyclopenta[c]pyrrolyl, each of the azetidinyl,
piperidinyl, pyrrolidinyl, azepanyl and
octahydrocyclopenta[c]pyrrolyl, each of which is optionally
substituted with one or more substituents independently selected
from the group consisting of (C.sub.1-C.sub.3)alkyl, --F,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, such as --CF.sub.3,
--CH.sub.2CF.sub.3 or --CH.sub.2CHF.sub.2.
[0089] In a ninth alternative embodiment, the compounds of the
present invention are represented by Structural Formula (II):
##STR00010##
[0090] wherein:
[0091] Y is selected from
--NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3),
##STR00011##
and --NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl, wherein the
--NH--S(O).sub.2--(C.sub.1-C.sub.6)alkyl in the group represented
by Y is optionally substituted with fluoro;
[0092] each R.sup.2 is independently selected from hydrogen and
--CH.sub.3; and
[0093] each R.sup.3 is independently selected from
(C.sub.1-C.sub.6)alkyl, and
--(C.sub.0-C.sub.1)alkylene-carbocyclyl; or
[0094] R.sup.2 and R.sup.3, taken together with the nitrogen atom
to which they are bound form a saturated heterocyclyl optionally
substituted with fluoro;
[0095] ring A represents a 4-7 membered saturated heterocyclyl;
[0096] R.sup.3' is hydrogen or methyl;
[0097] each carbocyclyl or heterocyclyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, (C.sub.1-C.sub.4)alkyl,
--O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.G)(R.sup.G);
[0098] each alkyl is optionally and independently substituted with
one or more substituents independently selected from fluoro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl; and
[0099] each R.sup.G is hydrogen or (C.sub.1-C.sub.3)alkyl, wherein
each alkyl group represented by R.sup.G is optionally and
independently substituted with one or more substituents
independently selected from fluoro, chloro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
[0100] Alternatively, for compounds of Structural Formula (II), the
heterocyclyl represented by ring A or --NR.sup.2R.sup.3 is
independently selected from the group consisting of azetidinyl,
morphinyl, piperazinyl, piperidinyl, pyrrolidinyl, azepanyl and
octahydrocyclopenta[c]pyrrolyl, each of which is optionally
substituted with fluoro. The remainder of the variables are as
described above for the ninth alternative embodiment.
[0101] Alternatively, for compounds of Structural Formula (II), the
carbocyclyl in the group represented by R.sup.3 is a
(C.sub.3-C.sub.6)cycloalkyl and the remainder of the variables are
as described in the ninth alternative embodiment.
[0102] In a tenth alternative embodiment, the compounds of the
present invention are represented by Structural Formula (III):
##STR00012##
[0103] wherein:
[0104] Y is selected from --NH.sub.2,
--NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3),
##STR00013##
--NH--C(O)-phenyl, and --NH--C(O)-thienyl, wherein:
[0105] ring A represents a 4-7 membered saturated heterocyclyl;
[0106] R.sup.3' is hydrogen or methyl;
[0107] the phenyl in the group represented by Y is optionally
substituted with --OCH.sub.3 or --N(CH.sub.3).sub.2;
[0108] each R.sup.2 is independently selected from hydrogen and
--CH.sub.3; and
[0109] each R.sup.3 is independently selected from
(C.sub.1-C.sub.6)alkyl, and
--(C.sub.0-C.sub.1)alkylene-carbocyclyl, wherein the
(C.sub.1-C.sub.6)alkyl is optionally substituted with fluoro or
--OCH.sub.3; or
[0110] R.sup.2 and R.sup.3, taken together with the nitrogen atom
to which they are bound form a saturated heterocyclyl optionally
substituted with fluoro or --OCH.sub.3;
[0111] each carbocyclyl or heterocyclyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, (C.sub.1-C.sub.4)alkyl,
--O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.G)(R.sup.G);
[0112] each alkyl is optionally and independently substituted with
one or more substituents independently selected from fluoro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl; and
[0113] each R.sup.G is hydrogen or (C.sub.1-C.sub.3)alkyl, wherein
each alkyl group represented by R.sup.G is optionally and
independently substituted with one or more substituents
independently selected from fluoro, chloro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
[0114] Alternatively, for compounds of Structural Formula (III)
described in the tenth alternative embodiment, the heterocyclyl
represented by ring A or --NR.sup.2R.sup.3 is independently
selected from the group consisting of azetidinyl, piperazinyl,
morpholinyl piperidinyl, pyrrolidinyl, azepanyl and
octahydrocyclopenta[c]pyrrolyl, each of which is optionally
substituted with fluoro or --OCH.sub.3; and the remainder of the
variables are as described in the tenth alternative embodiment.
[0115] In another embodiment, for compounds of Structural Formula
(III) described in the tenth alternative embodiment, the
carbocyclyl group represented by R.sup.3 is a
(C.sub.3-C.sub.6)cycloalkyl, and the remainder of the variables are
as described in the tenth alternative embodiment.
[0116] In a eleventh alternative embodiment, the compounds of the
present invention are represented by Structural Formula (IV):
##STR00014##
[0117] wherein:
[0118] Y is selected from --NH.sub.2,
--NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3), and
##STR00015##
wherein ring A represents a 4-7 membered saturated
heterocyclyl;
[0119] R.sup.3' is hydrogen or methyl;
[0120] each R.sup.2 is independently selected from hydrogen and
--CH.sub.3; and
[0121] each R.sup.3 is independently selected from
(C.sub.1-C.sub.6) alkyl, and
--(C.sub.0-C.sub.1)alkylene-carbocyclyl, wherein the
(C.sub.1-C.sub.6) alkyl is optionally substituted with fluoro or
--OCH.sub.3; or
[0122] R.sup.2 and R.sup.3, taken together with the nitrogen atom
to which they are bound form a saturated heterocyclyl optionally
substituted with fluoro or --OCH.sub.3;
[0123] each carbocyclyl or heterocyclyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, (C.sub.1-C.sub.4)alkyl,
--O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.G)(R.sup.G);
[0124] each alkyl is optionally and independently substituted with
one or more substituents independently selected from fluoro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl; and
[0125] each R.sup.G is hydrogen or (C.sub.1-C.sub.3)alkyl, wherein
each alkyl group represented by R.sup.G is optionally and
independently substituted with one or more substituents
independently selected from fluoro, chloro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
[0126] In one embodiment, for compounds of Structural Formula (IV)
described in the eleventh alternative embodiment, the heterocyclyl
represented by ring A or --NR.sup.2R.sup.3 is independently
selected from the group consisting of azetidinyl, piperidinyl,
pyrrolidinyl, azepanyl and octahydrocyclopenta[c]pyrrolyl, each of
which is optionally substituted with fluoro or --OCH.sub.3. The
remainder of the variables are as described above for the eleventh
alternative embodiment.
[0127] In another embodiment, for compounds of Structural Formula
(IV) described in the eleventh alternative embodiment, the
carbocyclyl in the group represented by R.sup.3 is a
(C.sub.3-C.sub.6)cycloalkyl and the remainder of the variables are
as described in the eleventh alternative embodiment.
[0128] In a twelfth alternative embodiment, the compounds of the
present invention is represented by Structural Formula (V):
##STR00016##
[0129] wherein:
[0130] Y is selected from hydrogen, --NH.sub.2,
--NH--C(O)--CH.sub.2--N(R.sup.2)(R.sup.3), and
##STR00017##
wherein ring A represents a 4-7 membered saturated
heterocyclyl;
[0131] R.sup.3' is hydrogen or methyl;
[0132] each R.sup.2 is independently selected from hydrogen and
--CH.sub.3; and
[0133] each R.sup.3 is independently selected from
(C.sub.1-C.sub.6) alkyl, and
--(C.sub.0-C.sub.1)alkylene-carbocyclyl, wherein the
(C.sub.1-C.sub.6) alkyl is optionally substituted with fluoro or
--OCH.sub.3; or
[0134] R.sup.2 and R.sup.3, taken together with the nitrogen atom
to which they are bound form a saturated heterocyclyl optionally
substituted with fluoro or --OCH.sub.3;
[0135] each carbocyclyl or heterocyclyl is optionally and
independently substituted with one or more substituents
independently selected from fluoro, (C.sub.1-C.sub.4)alkyl,
--O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, and
--N(R.sup.G)(R.sup.G);
[0136] each alkyl is optionally and independently substituted with
one or more substituents independently selected from fluoro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl;
[0137] each R.sup.G is hydrogen or (C.sub.1-C.sub.3)alkyl, wherein
each alkyl group represented by R.sup.G is optionally and
independently substituted with one or more substituents
independently selected from fluoro, chloro,
--O--(C.sub.1-C.sub.4)alkyl, and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
[0138] In one embodiment, for compounds of Structural Formula (V)
described in the twelfth alternative embodiment, the saturated
heterocyclyl represented by ring A or --NR.sup.2R.sup.3 is
independently is selected from the group consisting of azetidinyl,
piperidinyl, pyrrolidinyl, azepanyl and
octahydrocyclopenta[c]pyrrolyl. The remainder of the variables are
as described above for the twelfth alternative embodiment.
[0139] In another embodiment, for compounds of Structural Formula
(V) described in the twelfth alternative embodiment, the
carbocyclyl in the group represented by R.sup.3 is a
(C.sub.3-C.sub.6)cycloalkyl and the remainder of the variables are
as described in the twelfth alternative embodiment.
[0140] Exemplary compounds represented by Structural Formula (I)
are shown in Tables 1-4 below:
TABLE-US-00001 TABLE 1 Exemplary Compounds of Formula II
(X.dbd.OCH3) Table 1. Compound # Y 101 ##STR00018## 101
##STR00019## 102 ##STR00020## 103 ##STR00021## 104 ##STR00022## 105
##STR00023## 106 ##STR00024## 107 ##STR00025## 108 ##STR00026## 109
##STR00027## 110 ##STR00028## 111 ##STR00029## 112 ##STR00030## 113
##STR00031## 114 ##STR00032## 115 ##STR00033## 116 ##STR00034## 117
##STR00035## 118 ##STR00036## 119 ##STR00037## 120 ##STR00038## 121
##STR00039## 122 ##STR00040## 123 ##STR00041## 124 ##STR00042## 125
##STR00043## 126 ##STR00044## 127 ##STR00045## 128 ##STR00046## 129
##STR00047## 130 ##STR00048## 131 ##STR00049## 132 ##STR00050## 133
##STR00051## 134 ##STR00052## 135 ##STR00053## 136 ##STR00054## 137
##STR00055## 138 ##STR00056## 139 ##STR00057## 140 ##STR00058## 141
##STR00059## 142 ##STR00060## 143 ##STR00061## 144 ##STR00062## 145
##STR00063## 146 ##STR00064## 147 ##STR00065## 148 ##STR00066## 149
##STR00067## 150 ##STR00068## 151 ##STR00069## 152 ##STR00070## 153
##STR00071## 154 ##STR00072## 155 ##STR00073## 156 ##STR00074## 157
##STR00075## 158 ##STR00076## 159 ##STR00077## 160 ##STR00078## 161
##STR00079## 162 ##STR00080## 163 ##STR00081## 164 ##STR00082## 165
##STR00083## 166 ##STR00084## 167 ##STR00085## 168 ##STR00086## 169
##STR00087## 170 ##STR00088## 171 ##STR00089## 172 ##STR00090## 173
##STR00091## 174 ##STR00092## 175 ##STR00093## 176 ##STR00094## 177
##STR00095## 178 ##STR00096## 179 ##STR00097## 180 ##STR00098## 181
##STR00099## 182 ##STR00100## 183 ##STR00101## 184 ##STR00102## 185
##STR00103## 186 ##STR00104## 187 ##STR00105## 188 ##STR00106## 189
##STR00107## 190 ##STR00108## 191 ##STR00109## 192 ##STR00110## 193
##STR00111## 194 ##STR00112## 195 ##STR00113## 196 ##STR00114## 197
##STR00115## 198 ##STR00116## 199 ##STR00117## 200 ##STR00118## 201
##STR00119## 202 ##STR00120## 203 ##STR00121## 204 ##STR00122## 205
##STR00123## 206 ##STR00124##
TABLE-US-00002 TABLE 2 Exemplary Compounds of Formula III
(X.dbd.OCF.sub.3). Compound # Y 207 ##STR00125## 208 ##STR00126##
209 ##STR00127## 210 ##STR00128## 211 ##STR00129## 212 ##STR00130##
213 ##STR00131## 214 ##STR00132## 215 ##STR00133## 216 ##STR00134##
217 ##STR00135## 218 ##STR00136## 219 ##STR00137## 220 ##STR00138##
221 ##STR00139## 222 ##STR00140## 223 ##STR00141## 224 ##STR00142##
225 ##STR00143## 226 ##STR00144## 227 ##STR00145## 228 ##STR00146##
229 ##STR00147## 230 ##STR00148## 231 ##STR00149## 232 ##STR00150##
233 ##STR00151## 234 ##STR00152## 235 ##STR00153## 236 ##STR00154##
237 ##STR00155## 238 ##STR00156## 239 ##STR00157## 240 ##STR00158##
241 ##STR00159## 242 ##STR00160## 243 ##STR00161## 244 ##STR00162##
245 ##STR00163## 246 ##STR00164## 247 ##STR00165## 248 ##STR00166##
249 ##STR00167## 250 ##STR00168## 251 ##STR00169## 252 ##STR00170##
253 ##STR00171## 254 ##STR00172## 255 ##STR00173## 256 ##STR00174##
257 ##STR00175## 258 ##STR00176## 259 ##STR00177## 260 ##STR00178##
261 ##STR00179## 262 ##STR00180## 263 ##STR00181## 264 ##STR00182##
265 ##STR00183## 266 ##STR00184## 267 ##STR00185## 268
##STR00186##
TABLE-US-00003 TABLE 3 Exemplary Compounds of Formula IV
(X.dbd.CF.sub.3). Compound # Y 269 ##STR00187## 270 ##STR00188##
271 ##STR00189## 272 ##STR00190## 273 ##STR00191## 274 ##STR00192##
275 ##STR00193## 276 ##STR00194## 277 ##STR00195## 278 ##STR00196##
279 ##STR00197## 280 ##STR00198## 281 ##STR00199## 282 ##STR00200##
283 ##STR00201## 284 ##STR00202## 285 ##STR00203## 286 ##STR00204##
287 ##STR00205## 288 ##STR00206## 289 ##STR00207## 290 ##STR00208##
291 ##STR00209## 292 ##STR00210## 293 ##STR00211## 294 ##STR00212##
295 ##STR00213## 296 ##STR00214## 297 ##STR00215## 298 ##STR00216##
299 ##STR00217## 300 ##STR00218## 301 ##STR00219## 302 ##STR00220##
303 ##STR00221## 304 ##STR00222## 305 ##STR00223## 306 ##STR00224##
307 ##STR00225## 308 ##STR00226## 309 ##STR00227## 310 ##STR00228##
311 ##STR00229##
TABLE-US-00004 TABLE 4 Exemplary Compounds of Formula V (X.dbd.CN).
Compound # Y 312 ##STR00230## 313 ##STR00231## 314 ##STR00232## 315
##STR00233## 316 ##STR00234## 317 ##STR00235## 318 ##STR00236## 319
##STR00237## 320 ##STR00238## 321 ##STR00239## 322 ##STR00240## 323
##STR00241## 324 ##STR00242## 325 ##STR00243## 326 ##STR00244## 327
##STR00245## 328 ##STR00246## 329 ##STR00247## 330 ##STR00248## 331
##STR00249## 332 ##STR00250## 333 ##STR00251## 334 ##STR00252## 335
##STR00253##
Definitions
[0141] "Alkyl" means a saturated aliphatic branched or
straight-chain monovalent hydrocarbon radical having the specified
number of carbon atoms. Thus, "(C.sub.1-C.sub.6) alkyl" means a
radical having from 1-6 carbon atoms in a linear or branched
arrangement. "(C.sub.1-C.sub.6)alkyl" includes methyl, ethyl,
propyl, butyl, pentyl and hexyl.
[0142] "Alkylene" means a saturated aliphatic straight-chain
divalent hydrocarbon radical having the specified number of carbon
atoms. Thus, "(C.sub.1-C.sub.6) alkylene" means a divalent
saturated aliphatic radical having from 1-6 carbon atoms in a
linear arrangement, e.g., --[(CH.sub.2).sub.n], where n is an
integer from 1 to 6. "(C.sub.1-C.sub.6)alkylene" includes
methylene, ethylene, propylene, butylene, pentylene and
hexylene.
[0143] "Aryl" or "aromatic means an aromatic monocyclic or
polycyclic (e.g. bicyclic or tricyclic) carbocyclic ring system. In
one embodiment, "aryl" is a 6-12 membered monocylic or bicyclic
systems. Aryl systems include, but not limited to, phenyl,
naphthalenyl, fluorenyl, indenyl, azulenyl, and anthracenyl.
[0144] "Carbocyclyl" means a cyclic group with only ring carbon
atoms. "Carbocyclyl" includes 3-12 membered saturated or
unsaturated aliphatic cyclic hydrocarbon ring or 6-12 membered
aromatic ring. A carbocyclyl moiety can be monocyclic, fused
bicyclic, bridged bicyclic, spiro bicyclic, or polycyclic.
[0145] Monocyclic carbocyclyls are saturated or unsaturated
aliphatic cyclic hydrocarbon rings or aromatic hydrocarbon rings
having the specified number of carbon atoms. Monocyclic
carbocyclyls include cycloalkyl, cycloalkenyl, cycloalkynyl and
phenyl.
[0146] A fused bicyclic carbocyclyl has two rings which have two
adjacent ring atoms in common. The first ring is a monocyclic
carbocyclyl and the second ring is a monocylic carbocyclyl or a
moncyclic heterocyclyl.
[0147] A spiro bicyclic carbocyclyl has two rings which have only
one ring atom in common. The first ring is a monocyclic carbocyclyl
and the second ring is a monocylic carbocyclyl or a moncyclic
heterocyclyl.
[0148] A bridged bicyclic carbocyclyl has two rings which have
three or more adjacent ring atoms in common. The first ring is a
monocyclic carbocyclyl and the second ring is a monocylic
carbocyclyl or a moncyclic heterocyclyl.
[0149] Polycyclic carbocyclyls have more than two rings (e.g.,
three rings resulting in a tricyclic ring system) and adjacent
rings have at least one ring atom in common. The first ring is a
monocyclic carbocyclyl and the remainder of the ring structures are
monocyclic carbocyclyls or monocyclic heterocyclyls. Polycyclic
ring systems include fused, bridged and spiro ring systems. A fused
polycyclic ring system has at least two rings that have two
adjacent ring atoms in common. A spiro polycyclic ring system has
at least two rings that have only one ring atom in common. A
bridged polycyclic ring system has at least two rings that have
three or more adjacent ring atoms in common.
[0150] "Cycloalkyl" means a saturated aliphatic cyclic hydrocarbon
ring. Thus, "C.sub.3-C.sub.7 cycloalkyl" means a hydrocarbon
radical of a (3-7 membered) saturated aliphatic cyclic hydrocarbon
ring. A C.sub.3-C.sub.7 cycloalkyl includes, but is not limited to
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and
cycloheptyl.
[0151] "Cycloalkene" means an aliphatic cyclic hydrocarbon ring
having one or more double bonds in the ring.
[0152] "Cycloalkyne" means an aliphatic cyclic hydrocarbon ring
having one or more triple bonds in the ring.
[0153] "Heterocyclyl" means a cyclic 4-12 membered saturated or
unsaturated aliphatic ring containing 1, 2, 3, 4 or 5 heteroatoms
independently selected from N, O or S or a heteroaromatic ring.
When one heteroatom is S, it can be optionally mono- or
di-oxygenated (i.e. --S(O)-- or --S(O).sub.2--). The heterocyclyl
can be monocyclic, fused bicyclic, bridged bicyclic, spiro bicyclic
or polycyclic.
[0154] "Saturated heterocyclyl" means an aliphatic heterocyclyl
group without any degree of unsaturation (i.e., no double bond or
triple bond). It can be monocyclic, fused bicyclic, bridged
bicyclic, spiro bicyclic or polycyclic.
[0155] Examples of monocyclic saturated heterocyclyls include, but
are not limited to, azetidine, pyrrolidine, piperidine, piperazine,
azepane, hexahydropyrimidine, tetrahydrofuran, tetrahydropyran,
morpholine, thiomorpholine, thiomorpholine 1,1-dioxide,
tetrahydro-2H-1,2-thiazine, tetrahydro-2H-1,2-thiazine 1,1-dioxide,
isothiazolidine, isothiazolidine 1,1-dioxide. Examples of
heteroaromatic rings include, but are not limited to, furan,
thiophene, pyrrole, imidazole, pyrazole, oxazole, isoxazole,
thiazole, isothiazole, 1,2,3-triazole, 1,2,4-triazole,
1,3,4-oxadiazole, 1,2,5-thiadiazole, 1,2,5-thiadiazole 1-oxide,
1,2,5-thiadiazole 1,1-dioxide, 1,3,4-thiadiazole, pyridine,
pyridine-N-oxide, pyrazine, pyrimidine, pyridazine, 1,2,4-triazine,
1,3,5-triazine, and tetrazole.
[0156] A fused bicyclic heterocyclyl has two rings which have two
adjacent ring atoms in common. The first ring is a monocyclic
heterocyclyl and the second ring is a monocyclic carbocycle (such
as a cycloalkyl or phenyl) or a monocyclic heterocyclyl. For
example, the second ring is a (C.sub.3-C.sub.6)cycloalkyl, such as
cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. Alternatively,
the second ring is phenyl. Examples of fused bicyclic heterocyclyls
include, but are not limited to, octahydrocyclopenta[c]pyrrolyl,
indoline, isoindoline, 2,3-dihydro-1H-benzo[d]imidazole,
2,3-dihydrobenzo[d]oxazole, 2,3-dihydrobenzo[d]thiazole,
octahydrobenzo[d]oxazole, octahydro-1H-benzo[d]imidazole,
octahydrobenzo[d]thiazole, octahydrocyclopenta[c]pyrrole,
3-azabicyclo[3.1.0]hexane, 3-azabicyclo[3.2.0]heptane, indolizine,
indole, isoindole, indazole, benzimidazole, benzthiazole, purine,
quinoline, isoquinoline, cinnoline, phthalazine, quinazoline,
quinoxaline, 1,8-naphthyridine, and pteridine.
[0157] A Spiro bicyclic heterocyclyl has two rings which have only
one ring atom in common. The first ring is a monocyclic
heterocyclyl and the second ring is a monocyclic carbocycle (such
as a cycloalkyl or phenyl) or a monocyclic heterocyclyl. For
example, the second ring is a (C.sub.3-C.sub.6)cycloalkyl.
Alternatively, the second ring is phenyl. Example of spiro bicyclic
heterocyclyl includes, but are not limited to, azaspiro[4.4]nonane,
7-azaspiro[4.4]nonane, azasprio[4.5]decane, 8-azaspiro[4.5]decane,
azaspiro[5.5]undecane, 3-azaspiro[5.5]undecane and
3,9-diazaspiro[5.5]undecane.
[0158] A bridged bicyclic heterocyclyl has two rings which have
three or more adjacent ring atoms in common. The first ring is a
monocyclic heterocyclyl and the other ring is a monocyclic
carbocycle (such as a cycloalkyl or phenyl) or a monocyclic
heterocyclyl. Examples of bridged bicyclic heterocyclyls include,
but are not limited to, azabicyclo[3.3.1]nonane,
3-azabicyclo[3.3.1]nonane, azabicyclo[3.2.1]octane,
3-azabicyclo[3.2.1]octane, 6-azabicyclo[3.2.1]octane and
azabicyclo[2.2.2]octane, 2-azabicyclo[2.2.2]octane.
[0159] Polycyclic heterocyclyls have more than two rings, one of
which is a heterocyclyl (e.g., three rings resulting in a tricyclic
ring system) and adjacent rings having at least one ring atom in
common. Polycyclic ring systems include fused, bridged and spiro
ring systems. A fused polycyclic ring system has at least two rings
that have two adjacent ring atoms in common. A spiro polycyclic
ring system has at least two rings that have only one ring atom in
common. A bridged polycyclic ring system has at least two rings
that have three or more adjacent ring atoms in common.
[0160] Unless otherwise specified, a heterocyclyl or a carbocyclyl
can be optionally substituted with one or more (such as two, three,
four and five) substituents. Unless otherwise specified, suitable
substituents include (C.sub.1-C.sub.4)alkyl, halo, --OH,
(C.sub.1-C.sub.4)alkoxy, (C.sub.1-C.sub.4)alkylthio,
(C.sub.1-C.sub.4)alkylsulfinyl, (C.sub.1-C.sub.4)alkylsulfonyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl,
--N(R.sup.3)(R.sup.4), --CN, halo(C.sub.1-C.sub.4)alkyl, and
halo(C.sub.1-C.sub.4)alkoxy. Oxo (C.dbd.O) is also a suitable
substituent and examples of heterocyclic rings with an oxo
substituent include, but are not limited to, thiomorpholine
1-oxide, thiomorpholine 1,1-dioxide, tetrahydro-2H-1,2-thiazine
1,1-dioxide, and isothiazolidine 1,1-dioxide, pyrrolidin-2-one,
piperidin-2-one, piperazin-2-one, and morpholin-2-one. When the
heterocycle contains a ring nitrogen atom that is not connected to
an adjacent ring atom by a double bond, substituents at such ring
nitrogen atoms are also contemplated. Unless otherwise specified,
suitable substituents for a ring nitrogen atoms include alkyl (such
as -Me, or -Et) and acyl (e.g., --CHO, CH.sub.3CO--, and
CH.sub.3CH.sub.2CO--). In one embodiment, the substituents are
independently selected from the group consisting of --Cl, --F,
(C.sub.1-C.sub.4)alkyl, --OH, --O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, --C(O)--(C.sub.1-C.sub.4
alkyl), --C(O)-(fluoro-substituted-C.sub.1-C.sub.4 alkyl), and
--N(R.sup.2)(R.sup.2).
[0161] "Heteroaryl" or "heteroaromatic ring" means a 5-12 membered
monovalent heteroaromatic monocyclic or bicylic ring radical. A
herteroaryl contains 1, 2, 3 or 4 heteroatoms independently
selected from N, O, and S. Heteroaryls include, but are not limited
to pyrrole, imidazole, pyrazole, oxazole, isoxazole, thiazole,
isothiazole, 1,2,3-triazole, 1,2,4-triazole, 1,3,4-oxadiazole,
1,2,5-thiadiazole, 1,2,5-thiadiazole 1-oxide, 1,2,5-thiadiazole
1,1-dioxide, 1,3,4-thiadiazole, pyridine, pyrazine, pyrimidine,
pyridazine, 1,2,4-triazine, 1,3,5-triazine, and tetrazole. Bicyclic
heteroaryl rings include, but are not limited to, bicyclo[4.4.0]
and bicyclo[4.3.0] fused ring systems such as indolizine, indole,
isoindole, indazole, benzimidazole, benzthiazole, purine,
quinoline, isoquinoline, cinnoline, phthalazine, quinazoline,
quinoxaline, 1,8-naphthyridine, and pteridine.
[0162] "Hetero" refers to the replacement of at least one carbon
atom member in a ring system with at least one heteroatom selected
from N, S, and O. "Hetero" also refers to the replacement of at
least one carbon atom member in a acyclic system. A hetero ring
system or a hetero acyclic system may have 1, 2, 3 or 4 carbon atom
members replaced by a heteroatom.
[0163] "Halogen" used herein refers to fluorine, chlorine, bromine,
or iodine.
[0164] "Alkoxy" means an alkyl radical attached through an oxygen
linking atom. "(C.sub.1-C.sub.6)-alkoxy" includes methoxy, ethoxy,
propoxy, butoxy, pentoxy and hexoxy.
[0165] Haloalkyl and halocycloalkyl include mono, poly, and
perhaloalkyl groups where each halogen is independently selected
from fluorine, chlorine, and bromine. "Halogen" and "halo" are
interchangeably used herein and each refers to fluorine, chlorine,
bromine, or iodine.
[0166] "Fluoro" means --F.
[0167] As used herein, fluoro-substituted-(C.sub.1-C.sub.4)alkyl
means a (C.sub.1-C.sub.4)alkyl substituted with one or more --F
groups. Examples of fluoro-substituted-(C.sub.1-C.sub.4)alkyl
include, but are not limited to, --CF.sub.3, --CH.sub.2CF.sub.3,
--CH.sub.2CF.sub.2H, --CH.sub.2CH.sub.2F and
--CH.sub.2CH.sub.2CF.sub.3.
[0168] As used herein, "amino" include --NH.sub.2, monoalkylamino
and dialkylamino.
[0169] "Naturally occurring amino acid side chain moiety" refers to
any amino acid side chain moiety present in a natural amino
acid.
[0170] Unless otherwise specified, an alkyl group can be optionally
substituted with one or more (such as two, three, four and five)
substituents. Unless otherwise specified, the substituents are
independently selected from the group consisting of halogen (--Cl,
--F or --Br), alkyl, amino, --OH, alkoxy, haloalkyl,
halocycloalkyl, cycloalkl, aryl and heteroaryl. In one embodiment,
the substituents are independently selected from the group
consisting of --Cl, --F, (C.sub.1-C.sub.4)alkyl, --OH,
--O--(C.sub.1-C.sub.4)alkyl,
fluoro-substituted-(C.sub.1-C.sub.4)alkyl, --C(O)--(C.sub.1-C.sub.4
alkyl), --C(O)-(fluoro-substituted-C.sub.1-C.sub.4 alkyl), and
--N(R.sup.2)(R.sup.2). In another embodiment, the substituents are
independently selected from the group consisting of --F, --Cl,
--O--(C.sub.1-C.sub.4)alkyl and
fluoro-substituted-(C.sub.1-C.sub.4)alkyl.
[0171] Another embodiment of the present invention is a
pharmaceutical composition comprising one or more pharmaceutically
acceptable carrier and/or diluent and a compound disclosed herein
or a pharmaceutically acceptable salt thereof.
[0172] "Pharmaceutically acceptable carrier" and "pharmaceutically
acceptable diluent" means non-therapeutic components that are of
sufficient purity and quality for use in the formulation of a
composition of the invention that, when appropriately administered
to an animal or human, typically do not produce an adverse
reaction, and that are used as a vehicle for a drug substance (i.e.
a compound of the present invention).
[0173] Pharmaceutically acceptable salts of the compounds of the
present invention are also included. For example, an acid salt of a
compound of the present invention containing an amine or other
basic group can be obtained by reacting the compound with a
suitable organic or inorganic acid, resulting in pharmaceutically
acceptable anionic salt forms. Examples of anionic salts include
the acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate,
bromide, calcium edetate, camsylate, carbonate, chloride, citrate,
dihydrochloride, edetate, edisylate, estolate, esylate, fumarate,
glyceptate, gluconate, glutamate, glycollylarsanilate,
hexylresorcinate, hydrobromide, hydrochloride, hydroxynaphthoate,
iodide, isethionate, lactate, lactobionate, malate, maleate,
mandelate, mesylate, methylsulfate, mucate, napsylate, nitrate,
pamoate, pantothenate, phosphate/diphosphate, polygalacturonate,
salicylate, stearate, subacetate, succinate, sulfate, tannate,
tartrate, teoclate, tosylate, and triethiodide salts.
[0174] Salts of the compounds of the present invention containing a
carboxylic acid or other acidic functional group can be prepared by
reacting with a suitable base. Such a pharmaceutically acceptable
salt may be made with a base which affords a pharmaceutically
acceptable cation, which includes alkali metal salts (especially
sodium and potassium), alkaline earth metal salts (especially
calcium and magnesium), aluminum salts and ammonium salts, as well
as salts made from physiologically acceptable organic bases such as
trimethylamine, triethylamine, morpholine, pyridine, piperidine,
picoline, dicyclohexylamine, N,N'-dibenzylethylenediamine,
2-hydroxyethylamine, bis-(2-hydroxyethyl)amine,
tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine,
dehydroabietylamine, N,N'-bisdehydroabietylamine, glucamine,
N-methylglucamine, collidine, quinine, quinoline, and basic amino
acids such as lysine and arginine.
[0175] The invention also includes various isomers and mixtures
thereof. Certain of the compounds of the present invention may
exist in various stereoisomeric forms. Stereoisomers are compounds
which differ only in their spatial arrangement. Enantiomers are
pairs of stereoisomers whose mirror images are not superimposable,
most commonly because they contain an asymmetrically substituted
carbon atom that acts as a chiral center. "Enantiomer" means one of
a pair of molecules that are minor images of each other and are not
superimposable. Diastereomers are stereoisomers that are not
related as mirror images, most commonly because they contain two or
more asymmetrically substituted carbon atoms. "R" and "S" represent
the configuration of substituents around one or more chiral carbon
atoms. When a chiral center is not defined as R or S, either a pure
enantiomer or a mixture of both configurations is present.
[0176] "Racemate" or "racemic mixture" means a compound of
equimolar quantities of two enantiomers, wherein such mixtures
exhibit no optical activity; i.e., they do not rotate the plane of
polarized light.
[0177] The compounds of the invention may be prepared as individual
isomers by either isomer-specific synthesis or resolved from an
isomeric mixture. Conventional resolution techniques include
forming the salt of a free base of each isomer of an isomeric pair
using an optically active acid (followed by fractional
crystallization and regeneration of the free base), forming the
salt of the acid form of each isomer of an isomeric pair using an
optically active amine (followed by fractional crystallization and
regeneration of the free acid), forming an ester or amide of each
of the isomers of an isomeric pair using an optically pure acid,
amine or alcohol (followed by chromatographic separation and
removal of the chiral auxiliary), or resolving an isomeric mixture
of either a starting material or a final product using various well
known chromatographic methods.
[0178] When the stereochemistry of a disclosed compound is named or
depicted by structure, the named or depicted stereoisomer is at
least 60%, 70%, 80%, 90%, 99% or 99.9% by weight pure relative to
the other stereoisomers. When a single enantiomer is named or
depicted by structure, the depicted or named enantiomer is at least
60%, 70%, 80%, 90%, 99% or 99.9% by weight optically pure. Percent
optical purity by weight is the ratio of the weight of the
enantiomer that is present divided by the combined weight of the
enantiomer that is present and the weight of its optical
isomer.
[0179] The present invention also provides a method of treating or
preventing a subject with a tetracycline-responsive disease or
disorder comprising administering to the subject an effective
amount of a compound of the present invention or a pharmaceutically
acceptable salt thereof.
[0180] "Tetracycline-responsive disease or disorder" refers to a
disease or disorder that can be treated, prevented, or otherwise
ameliorated by the administration of a tetracycline compound of the
present invention. Tetracycline-responsive disease or disorder
includes infections, cancer, inflammatory disorders, autoimmune
disease, arteriosclerosis, corneal ulceration, emphysema,
arthritis, osteoporosis, osteoarthritis, multiple sclerosis,
osteosarcoma, osteomyelitis, bronchiectasis, chronic pulmonary
obstructive disease, skin and eye diseases, periodontitis,
osteoporosis, rheumatoid arthritis, ulcerative colitis,
prostatitis, tumor growth and invasion, metastasis, diabetes,
diabetic proteinuria, panbronchiolitis; aortic or vascular
aneurysms, skin tissue wounds, dry eye, bone, cartilage
degradation, malaria, senescence, diabetes, vascular stroke,
neurodegenerative disorders, cardiac disease, juvenile diabetes,
acute and chronic bronchitis, sinusitis, and respiratory
infections, including the common cold; Wegener's granulomatosis;
neutrophilic dermatoses and other inflammatory diseases such as
dermatitis herpetiformis, leukocytoclastic vasculitis, bullous
lupus erythematosus, pustular psoriasis, erythema elevatum
diutinum; vitiligo; discoid lupus erythematosus; pyoderma
gangrenosum; pustular psoriasis; blepharitis, or meibomianitis;
Alzheimer's disease; degenerative maculopathy; acute and chronic
gastroenteritis and colitis; acute and chronic cystitis and
urethritis; acute and chronic dermatitis; acute and chronic
conjunctivitis; acute and chronic serositis; uremic pericarditis;
acute and chronic cholecystis; cystic fibrosis, acute and chronic
vaginitis; acute and chronic uveitis; drug reactions; insect bites;
burns and sunburn, bone mass disorder, acute lung injury, chronic
lung disorders, ischemia, stroke or ischemic stroke, skin wound,
aortic or vascular aneurysm, diabetic retinopathy, hemorrhagic
stroke, angiogenesis, and other states for which tetracycline
compounds have been found to be active (see, for example, U.S. Pat.
Nos. 5,789,395; 5,834,450; 6,277,061 and 5,532,227, each of which
is expressly incorporated herein by reference).
[0181] In addition, a method to treat any disease or disease state
that could benefit from modulating the expression and/or function
of nitric oxide, metalloproteases, proinflammatory mediators and
cytokines, reactive oxygen species, components of the immune
response, including chemotaxis, lymphocyte transformation, delayed
hypersensitivity, antibody production, phagocytosis, and oxidative
metabolism of phagocytes. A method to treat any disease or disease
state that could benefit from modulating the expression and/or
function of C-reactive protein, signaling pathways (e.g., FAK
signaling pathway), and/or augment the expression of COX-2 and PGE2
production is covered. A method to treat any disease or disease
state that could benefit from inhibition of neovascularization is
covered.
[0182] Compounds of the invention can be used to prevent or treat
important mammalian and veterinary diseases such as diarrhea,
urinary tract infections, infections of skin and skin structure,
ear, nose and throat infections, wound infection, mastitis and the
like. In addition, methods for treating neoplasms using
tetracycline compounds of the invention are also included (van der
Bozert et al., Cancer Res., 48: 6686-6690 (1988)).
[0183] Infections that can be treated using compounds of the
invention or a pharmaceutically acceptable salt thereof include,
but are not limited to, skin infections, GI infections, urinary
tract infections, genito-urinary infections, respiratory tract
infections, sinuses infections, middle ear infections, systemic
infections, cholera, influenza, bronchitis, acne, malaria, sexually
transmitted disease including syphilis and gonorrhea, Legionnaires'
disease, Lyme disease, Rocky Mountain spotted fever, Q fever,
typhus, bubonic plague, gas gangrene, hospital acquired infections,
leptospirosis, whooping cough, anthrax and infections caused by the
agents responsible for lymphogranuloma venereum, inclusion
conjunctivitis, or psittacosis. Infections can be bacterial,
fungal, parasitic and viral infections (including those which are
resistant to other tetracycline compounds).
[0184] In one embodiment, the infection can be caused bacteria. In
another embodiment, the infection is caused by a Gram-positive
bacteria. In a specific aspect of this embodiment, the infection is
caused by a Gram-positive bacterium selected from Staphylococcus
spp., Streptococcus spp., Propionibacterium spp., Enterococcus
spp., Bacillus spp., Corynebacterium spp., Nocardia spp.,
Clostridium spp., Actinobacteria spp., and Listeria spp.
[0185] In another embodiment, the infection is caused by a
Gram-negative bacterium. In one aspect of this embodiment, the
infection is caused by a proteobacteria (e.g., Betaproteobacteria
and Gammaproteobacteria), including Escherichia coli, Salmonella,
Shigella, other Enterobacteriaceae, Pseudomonas, Moraxella,
Helicobacter, Stenotrophomonas, Bdellovibrio, acetic acid bacteria,
Legionella or alpha-proteobacteria such as Wolbachia. In another
aspect, the infection is caused by a Gram-negative bacteria
selected from cyanobacteria, spirochaetes, green sulfur or green
non-sulfur bacteria. In a specific aspect of this embodiment, the
infection is caused by a Gram-negative bacteria selected from
Enterobactericeae (e.g., E. coli, Klebsiella pneumonia including
those containing extended-spectrum .beta.-lactamases and/or
carbapenemases), Bacteroidaceae (e.g., Bacteroides fragilis),
Vibrionaceae (Vibrio cholerae), Pasteurellae (e.g., Haemophilus
influenza), Pseudomonadaceae (e.g., Pseudomonas aeruginosa),
Neisseriaceae (e.g. Neisseria meningitidis), Rickettsiae,
Moraxellaceae (e.g., Moraxella catarrhalis), any species of
Proteeae, Acinetobacter spp., Helicobacter spp., and Campylobacter
spp.
[0186] In a particular embodiment, the infection is caused by
Gram-negative bacterium selected from the group consisting of
Enterobactericeae (e.g., E. coli, Klebsiella pneumoniae),
Pseudomonas, and Acinetobacter spp.
[0187] In another embodiment, the infection is caused by an
organism selected from the group consisting of K. pneumoniae,
Salmonella, E. hirae, A. baumanii, M. catarrhalis, H. influenzae,
P. aeruginosa, E. faecium, E. coli, S. aureus, and E. faecalis.
[0188] In another embodiment, the infection is caused by an
organism selected from the group consisting of rickettsiae,
chlamydiae, Legionella spp. and Mycoplasma spp.
[0189] In another embodiment, the infection is caused by an
organism resistant to tetracycline or any member of first and
second generation of tetracycline antibiotics (e.g., doxycycline or
minocycline).
[0190] In another embodiment, the infection is caused by an
organism resistant to methicillin.
[0191] In another embodiment, the infection is caused by an
organism resistant to vancomycin.
[0192] In another embodiment, the infection is caused by an
organism resistant to a quinolone or fluoroquinolone.
[0193] In another embodiment, the infection is caused by an
organism resistant to tigecycline.
[0194] In another embodiment, the infection is caused by a
multidrug-resistant pathogen (having intermediate or full
resistance to any two or more antibiotics). In another embodiment
the infection is a Bacillus anthracis infection. "Bacillus
anthracis infection" includes any state, diseases, or disorders
caused or which result from exposure or alleged exposure to
Bacillus anthracis or another member of the Bacillus cereus group
of bacteria. In another embodiment, the infection is caused by
Bacillus anthracis (anthrax), Yersinia pestis (plague), or
Francisella tularensis (tularemia).
[0195] In yet another embodiment, the infection can be caused by
more than one organism described above. Examples of such infections
include, but are not limited to, intra-abdominal infections (often
a mixture of a gram-negative species like E. coli and an anaerobe
like B. fragilis), diabetic foot (various combinations of
Streptococcus, Serratia, Staphylococcus and Enterococcus spp.,
anaerobes (S. E. Dowd, et al., PloS one 2008; 3:e3326) and
respiratory disease (especially in patients that have chronic
infections like cystic fibrosis--e.g., S. aureus plus P. aeruginosa
or H. influenza, atypical pathogens), wounds and abscesses (various
gram-negative and gram-positive bacteria, notably MSSA/MRSA,
coagulase-negative staphylococci, enterococci, Acinetobacter, P.
aeruginosa, E. coli, B. fragilis), and bloodstream infections (13%
were polymicrobial (H. Wisplinghoff, et al., Clin. Infect. Dis.
2004; 39:311-317)).
[0196] In a further embodiment, the tetracycline responsive disease
or disorder is not a bacterial infection. In another embodiment,
the tetracycline compounds of the invention are essentially
non-antibacterial. For example, non-antibacterial compounds of the
invention may have MIC values greater than about 4 .mu.g/ml (as
measured by assays known in the art and/or the assay given in
Example 14. In another embodiment, the tetracycline compounds of
the invention have both antibacterial and non-antibacterial
effects.
[0197] Tetracycline responsive disease or disorder also includes
diseases or disorders associated with inflammatory process
associated states (IPAS). The term "inflammatory process associated
state" includes states in which inflammation or inflammatory
factors (e.g., matrix metalloproteinases (MMPs), nitric oxide (NO),
TNF, interleukins, plasma proteins, cellular defense systems,
cytokines, lipid metabolites, proteases, toxic radicals, adhesion
molecules, etc.) are involved or are present in an area in aberrant
amounts, e.g., in amounts which may be advantageous to alter, e.g.,
to benefit the subject. The inflammatory process is the response of
living tissue to damage. The cause of inflammation may be due to
physical damage, chemical substances, micro-organisms, tissue
necrosis, cancer or other agents. Acute inflammation is
short-lasting, lasting only a few days. If it is longer lasting
however, then it may be referred to as chronic inflammation.
[0198] IPASs include inflammatory disorders. Inflammatory disorders
are generally characterized by heat, redness, swelling, pain and
loss of function. Examples of causes of inflammatory disorders
include, but are not limited to, microbial infections (e.g.,
bacterial and fungal infections), physical agents (e.g., burns,
radiation, and trauma), chemical agents (e.g., toxins and caustic
substances), tissue necrosis and various types of immunologic
reactions.
[0199] Examples of inflammatory disorders can be treated using the
compounds of the invention or a pharmaceutically acceptable salt
thereof include, but are not limited to, osteoarthritis, rheumatoid
arthritis, acute and chronic infections (bacterial and fungal,
including diphtheria and pertussis); acute and chronic bronchitis,
sinusitis, and upper respiratory infections, including the common
cold; acute and chronic gastroenteritis and colitis; inflammatory
bowel disorder; acute and chronic cystitis and urethritis;
vasculitis; sepsis; nephritis; pancreatitis; hepatitis; lupus;
inflammatory skin disorders including, for example, eczema,
dermatitis, psoriasis, pyoderma gangrenosum, acne rosacea, and
acute and chronic dermatitis; acute and chronic conjunctivitis;
acute and chronic serositis (pericarditis, peritonitis, synovitis,
pleuritis and tendinitis); uremic pericarditis; acute and chronic
cholecystis; acute and chronic vaginitis; acute and chronic
uveitis; drug reactions; insect bites; burns (thermal, chemical,
and electrical); and sunburn.
[0200] IPASs also include matrix metalloproteinase associated
states (MMPAS). MMPAS include states characterized by aberrant
amounts of MMPs or MMP activity.
[0201] Examples of matrix metalloproteinase associated states
("MMPAS's") can be treated using compounds of the invention or a
pharmaceutically acceptable salt thereof, include, but are not
limited to, arteriosclerosis, corneal ulceration, emphysema,
osteoarthritis, multiple sclerosis (Liedtke et al., Ann. Neurol.
1998, 44: 35-46; Chandler et al., J. Neuroimmunol. 1997, 72:
155-71), osteosarcoma, osteomyelitis, bronchiectasis, chronic
pulmonary obstructive disease, skin and eye diseases,
periodontitis, osteoporosis, rheumatoid arthritis, ulcerative
colitis, inflammatory disorders, tumor growth and invasion
(Stetler-Stevenson et al., Annu. Rev. Cell Biol. 1993, 9: 541-73;
Tryggvason et al., Biochim. Biophys. Acta 1987, 907: 191-217; Li et
al., Mol. Carcillog. 1998, 22: 84-89)), metastasis, acute lung
injury, stroke, ischemia, diabetes, aortic or vascular aneurysms,
skin tissue wounds, dry eye, bone and cartilage degradation
(Greenwald et al., Bone 1998,22: 33-38; Ryan et al., Curr. Op.
Rheumatol. 1996, 8: 238-247). Other MMPAS include those described
in U.S. Pat. Nos. 5,459,135; 5,321,017; 5,308,839; 5,258,371;
4,935,412; 4,704,383, 4,666,897, and RE 34,656, incorporated herein
by reference in their entirety.
[0202] In a further embodiment, the IPAS includes disorders
described in U.S. Pat. Nos. 5,929,055; and 5,532,227, incorporated
herein by reference in their entirety.
[0203] Tetracycline responsive disease or disorder also includes
diseases or disorders associated with NO associated states. The
term "NO associated states" includes states which involve or are
associated with nitric oxide (NO) or inducible nitric oxide
synthase (iNOS). NO associated state includes states which are
characterized by aberrant amounts of NO and/or iNOS. Preferably,
the NO associated state can be treated by administering
tetracycline compounds of the invention. The disorders, diseases
and states described in U.S. Pat. Nos. 6,231,894; 6,015,804;
5,919,774; and 5,789,395 are also included as NO associated states.
The entire contents of each of these patents are hereby
incorporated herein by reference.
[0204] Examples of diseases or disorders associated with NO
associated states can be treated using the compounds of the present
invention or a pharmaceutically acceptable salt thereof include,
but are not limited to, malaria, senescence, diabetes, vascular
stroke, neurodegenerative disorders (Alzheimer's disease and
Huntington's disease), cardiac disease (reperfusion-associated
injury following infarction), juvenile diabetes, inflammatory
disorders, osteoarthritis, rheumatoid arthritis, acute, recurrent
and chronic infections (bacterial, viral and fungal); acute and
chronic bronchitis, sinusitis, and respiratory infections,
including the common cold; acute and chronic gastroenteritis and
colitis; acute and chronic cystitis and urethritis; acute and
chronic dermatitis; acute and chronic conjunctivitis; acute and
chronic serositis (pericarditis, peritonitis, synovitis, pleuritis
and tendonitis); uremic pericarditis; acute and chronic
cholecystis; cystic fibrosis, acute and chronic vaginitis; acute
and chronic uveitis; drug reactions; insect bites; burns (thermal,
chemical, and electrical); and sunburn.
[0205] In another embodiment, the tetracycline responsive disease
or disorder is cancer. Examples of cancers that can be treated
using the compounds of the invention or a pharmaceutically
acceptable salt thereof include all solid tumors, i.e., carcinomas
e.g., adenocarcinomas, and sarcomas. Adenocarcinomas are carcinomas
derived from glandular tissue or in which the tumor cells form
recognizable glandular structures. Sarcomas broadly include tumors
whose cells are embedded in a fibrillar or homogeneous substance
like embryonic connective tissue. Examples of carcinomas which may
be treated using the methods of the invention include, but are not
limited to, carcinomas of the prostate, breast, ovary, testis,
lung, colon, and breast. The methods of the invention are not
limited to the treatment of these tumor types, but extend to any
solid tumor derived from any organ system. Examples of treatable
cancers include, but are not limited to, colon cancer, bladder
cancer, breast cancer, melanoma, ovarian carcinoma, prostate
carcinoma, lung cancer, and a variety of other cancers as well. The
methods of the invention also cause the inhibition of cancer growth
in adenocarcinomas, such as, for example, those of the prostate,
breast, kidney, ovary, testes, and colon. In one embodiment, the
cancers treated by methods of the invention include those described
in U.S. Pat. Nos. 6,100,248; 5,843,925; 5,837,696; or 5,668,122,
incorporated herein by reference in their entirety.
[0206] Alternatively, the tetracycline compounds may be useful for
preventing or reducing the likelihood of cancer recurrence, for
example, to treat residual cancer following surgical resection or
radiation therapy. The tetracycline compounds useful according to
the invention are especially advantageous as they are substantially
non-toxic compared to other cancer treatments.
[0207] In a further embodiment, the compounds of the invention are
administered in combination with standard cancer therapy, such as,
but not limited to, chemotherapy.
[0208] Examples of tetracycline responsive states can be treated
using the compounds of the invention or a pharmaceutically
acceptable salt thereof also include neurological disorders which
include both neuropsychiatric and neurodegenerative disorders, but
are not limited to, such as Alzheimer's disease, dementias related
to Alzheimer's disease (such as Pick's disease), Parkinson's and
other Lewy diffuse body diseases, senile dementia, Huntington's
disease, Gilles de la Tourette's syndrome, multiple sclerosis,
amyotrophic lateral sclerosis (ALS), progressive supranuclear
palsy, epilepsy, and Creutzfeldt-Jakob disease; autonomic function
disorders such as hypertension and sleep disorders, and
neuropsychiatric disorders, such as depression, schizophrenia,
schizoaffective disorder, Korsakoffs psychosis, mania, anxiety
disorders, or phobic disorders; learning or memory disorders, e.
g., amnesia or age-related memory loss, attention deficit disorder,
dysthymic disorder, major depressive disorder, mania,
obsessive-compulsive disorder, psychoactive substance use
disorders, anxiety, phobias, panic disorder, as well as bipolar
affective disorder, e.g., severe bipolar affective (mood) disorder
(BP-1), bipolar affective neurological disorders, e.g., migraine
and obesity.
[0209] Further neurological disorders include, for example, those
listed in the American Psychiatric Association's Diagnostic and
Statistical manual of Mental Disorders (DSM), the most current
version of which is incorporated herein by reference in its
entirety.
[0210] In another embodiment, the tetracycline responsive disease
or disorder is diabetes. Diabetes that can be treated using the
compounds of the invention or a pharmaceutically acceptable salt
thereof include, but are not limited to, juvenile diabetes,
diabetes mellitus, diabetes type I, or diabetes type II. In a
further embodiment, protein glycosylation is not affected by the
administration of the tetracycline compounds of the invention. In
another embodiment, the tetracycline compound of the invention is
administered in combination with standard diabetic therapies, such
as, but not limited to insulin therapy.
[0211] In another embodiment, the tetracycline responsive disease
or disorder is a bone mass disorder. Bone mass disorders that can
be treated using the compounds of the invention or a
pharmaceutically acceptable salt thereof include disorders where a
subjects bones are disorders and states where the formation, repair
or remodeling of bone is advantageous. For examples bone mass
disorders include osteoporosis (e.g., a decrease in bone strength
and density), bone fractures, bone formation associated with
surgical procedures (e.g., facial reconstruction), osteogenesis
imperfecta (brittle bone disease), hypophosphatasia, Paget's
disease, fibrous dysplasia, osteopetrosis, myeloma bone disease,
and the depletion of calcium in bone, such as that which is related
to primary hyperparathyroidism. Bone mass disorders include all
states in which the formation, repair or remodeling of bone is
advantageous to the subject as well as all other disorders
associated with the bones or skeletal system of a subject which can
be treated with the tetracycline compounds of the invention. In a
further embodiment, the bone mass disorders include those described
in U.S. Pat. Nos. 5,459,135; 5,231,017; 5,998,390; 5,770,588; RE
34,656; 5,308,839; 4,925,833; 3,304,227; and 4,666,897, each of
which is hereby incorporated herein by reference in its
entirety.
[0212] In another embodiment, the tetracycline responsive disease
or disorder is acute lung injury. Acute lung injuries that can be
treated using the compounds of the invention or a pharmaceutically
acceptable salt thereof include adult respiratory distress syndrome
(ARDS), post-pump syndrome (PPS), and trauma. Trauma includes any
injury to living tissue caused by an extrinsic agent or event.
Examples of trauma include, but are not limited to, crush injuries,
contact with a hard surface, or cutting or other damage to the
lungs.
[0213] The tetracycline responsive disease or disorders of the
invention also include chronic lung disorders. Examples of chronic
lung disorders that can be treated using the compounds of the
invention or a pharmaceutically acceptable salt thereof include,
but are not limited, to asthma, cystic fibrosis, chronic
obstructive pulmonary disease (COPD), and emphysema. In a further
embodiment, the acute and/or chronic lung disorders that can be
treated using the compounds of the invention or a pharmaceutically
acceptable salt thereof include those described in U.S. Pat. No.
5,977,091; 6,043,231; 5,523,297; and 5,773,430, each of which is
hereby incorporated herein by reference in its entirety.
[0214] In yet another embodiment, the tetracycline responsive
disease or disorder is ischemia, stroke, or ischemic stroke.
[0215] In a further embodiment, the tetracycline compounds of the
invention or a pharmaceutically acceptable salt thereof can be used
to treat such disorders as described above and in U.S. Pat. Nos.
6,231,894; 5,773,430; 5,919,775 and 5,789,395, incorporated herein
by reference.
[0216] In still a further embodiment, the tetracycline compounds of
the invention or a pharmaceutically acceptable salt thereof can be
used to treat pain, for example, inflammatory, nociceptive or
neuropathic pain. The pain can be either acute or chronic.
[0217] In another embodiment, the tetracycline responsive disease
or disorder is a skin wound. The invention also provides a method
for improving the healing response of the epithelialized tissue
(e.g., skin, mucosae) to acute traumatic injury (e.g., cut, burn,
scrape, etc.). The method includes using a tetracycline compound of
the invention or a pharmaceutically acceptable salt thereof to
improve the capacity of the epithelialized tissue to heal acute
wounds. The method may increase the rate of collagen accumulation
of the healing tissue. The method may also decrease the proteolytic
activity in the epithelialized tissue by decreasing the
collagenolytic and/or gellatinolytic activity of MMPs. In a further
embodiment, the tetracycline compound of the invention or a
pharmaceutically acceptable salt thereof is administered to the
surface of the skin (e. g., topically). In a further embodiment,
the tetracycline compound of the invention or a pharmaceutically
acceptable salt thereof is used to treat a skin wound, and other
such disorders as described in, for example, U.S. Pat. Nos.
5,827,840; 4,704,383; 4,935,412; 5,258,371; 5,308,839, 5,459,135;
5,532,227; and 6,015,804; each of which is incorporated herein by
reference in its entirety.
[0218] In yet another embodiment, the tetracycline responsive
disease or disorder is an aortic or vascular aneurysm in vascular
tissue of a subject (e.g., a subject having or at risk of having an
aortic or vascular aneurysm, etc.). The tetracycline compound or a
pharmaceutically acceptable salt thereof may be effective to reduce
the size of the vascular aneurysm or it may be administered to the
subject prior to the onset of the vascular aneurysm such that the
aneurysm is prevented. In one embodiment, the vascular tissue is an
artery, e.g., the aorta, e.g., the abdominal aorta. In a further
embodiment, the tetracycline compounds of the invention are used to
treat disorders described in U.S. Pat. Nos. 6,043,225 and
5,834,449, incorporated herein by reference in their entirety.
[0219] The compounds of the invention or a pharmaceutically
acceptable salt thereof can be used alone or in combination with
one or more therapeutic agent in the methods of the invention
disclosed herein.
[0220] The language "in combination with" another therapeutic agent
or treatment includes co-administration of the tetracycline
compound and with the other therapeutic agent or treatment as
either a single combination dosage form or as multiple, separate
dosage forms, administration of the tetracycline compound first,
followed by the other therapeutic agent or treatment and
administration of the other therapeutic agent or treatment first,
followed by the tetracycline compound.
[0221] The other therapeutic agent may be any agent that is known
in the art to treat, prevent, or reduce the symptoms of a
tetracycline-responsive disease or disorder. The choice of
additional therapeutic agent(s) is based upon the particular
tetracycline-responsive disease or disorder being treated. Such
choice is within the knowledge of a treating physician.
Furthermore, the other therapeutic agent may be any agent of
benefit to the patient when administered in combination with the
administration of a tetracycline compound.
[0222] As used herein, the term "subject" means a mammal in need of
treatment or prevention, e.g., companion animals (e.g., dogs, cats,
and the like), farm animals (e.g., cows, pigs, horses, sheep, goats
and the like) and laboratory animals (e.g., rats, mice, guinea pigs
and the like). Typically, the subject is a human in need of the
specified treatment.
[0223] As used herein, the term "treating" or `treatment" refers to
obtaining desired pharmacological and/or physiological effect. The
effect can include achieving, partially or substantially, one or
more of the following results: partially or totally reducing the
extent of the disease, disorder or syndrome; ameliorating or
improving a clinical symptom or indicator associated with the
disorder; delaying, inhibiting or decreasing the likelihood of the
progression of the disease, disorder or syndrome.
[0224] As used herein, "preventing" or "prevention" refers to
reducing the likelihood of the onset or development of disease,
disorder or syndrome.
[0225] "Effective amount" means that amount of active compound
agent that elicits the desired biological response in a subject. In
one embodiment, the effective amount of a compound of the invention
is from about 0.01 mg/kg/day to about 1000 mg/kg/day, from about
0.1 mg/kg/day to about 100 mg/kg/day, or from about 0.5 mg/kg/day
to about 50 mg/kg/day.
[0226] The invention further includes the process for making the
composition comprising mixing one or more of the present compounds
and an optional pharmaceutically acceptable carrier; and includes
those compositions resulting from such a process, which process
includes conventional pharmaceutical techniques.
[0227] The compositions of the invention include ocular, oral,
nasal, transdermal, topical with or without occlusion, intravenous
(both bolus and infusion), inhalable, and injection
(intraperitoneally, subcutaneously, intramuscularly,
intratumorally, or parenterally) formulations. The composition may
be in a dosage unit such as a tablet, pill, capsule, powder,
granule, liposome, ion exchange resin, sterile ocular solution, or
ocular delivery device (such as a contact lens and the like
facilitating immediate release, timed release, or sustained
release), parenteral solution or suspension, metered aerosol or
liquid spray, drop, ampoule, auto-injector device, or suppository;
for administration ocularly, orally, intranasally, sublingually,
parenterally, or rectally, or by inhalation or insufflation.
[0228] Compositions of the invention suitable for oral
administration include solid forms such as pills, tablets, caplets,
capsules (each including immediate release, timed release, and
sustained release formulations), granules and powders; and, liquid
forms such as solutions, syrups, elixirs, emulsions, and
suspensions. Forms useful for ocular administration include sterile
solutions or ocular delivery devices. Forms useful for parenteral
administration include sterile solutions, emulsions, and
suspensions.
[0229] The compositions of the invention may be administered in a
form suitable for once-weekly or once-monthly administration. For
example, an insoluble salt of the active compound may be adapted to
provide a depot preparation for intramuscular injection (e.g., a
decanoate salt) or to provide a solution for ophthalmic
administration.
[0230] The dosage form containing the composition of the invention
contains an effective amount of the active ingredient necessary to
provide a therapeutic effect. The composition may contain from
about 5,000 mg to about 0.5 mg (preferably, from about 1,000 mg to
about 0.5 mg) of a compound of the invention or salt form thereof
and may be constituted into any form suitable for the selected mode
of administration. The composition may be administered about 1 to
about 5 times per day. Daily administration or post-periodic dosing
may be employed.
[0231] For oral administration, the composition is preferably in
the form of a tablet or capsule containing, e.g., 500 to 0.5
milligrams of the active compound. Dosages will vary depending on
factors associated with the particular patient being treated (e.g.,
age, weight, diet, and time of administration), the severity of the
condition being treated, the compound being employed, the mode of
administration, and the strength of the preparation.
[0232] The oral composition is preferably formulated as a
homogeneous composition, wherein the active ingredient is dispersed
evenly throughout the mixture, which may be readily subdivided into
dosage units containing equal amounts of a compound of the
invention. Preferably, the compositions are prepared by mixing a
compound of the invention (or pharmaceutically acceptable salt
thereof) with one or more optionally present pharmaceutical
carriers (such as a starch, sugar, diluent, granulating agent,
lubricant, glidant, binding agent, and disintegrating agent), one
or more optionally present inert pharmaceutical excipients (such as
water, glycols, oils, alcohols, flavoring agents, preservatives,
coloring agents, and syrup), one or more optionally present
conventional tableting ingredients (such as corn starch, lactose,
sucrose, sorbitol, talc, stearic acid, magnesium stearate,
dicalcium phosphate, and any of a variety of gums), and an optional
diluent (such as water).
[0233] Binder agents include starch, gelatin, natural sugars (e.g.,
glucose and beta-lactose), corn sweeteners and natural and
synthetic gums (e.g., acacia and tragacanth). Disintegrating agents
include starch, methyl cellulose, agar, and bentonite.
[0234] Tablets and capsules represent an advantageous oral dosage
unit form. Tablets may be sugarcoated or filmcoated using standard
techniques. Tablets may also be coated or otherwise compounded to
provide a prolonged, control-release therapeutic effect. The dosage
form may comprise an inner dosage and an outer dosage component,
wherein the outer component is in the form of an envelope over the
inner component. The two components may further be separated by a
layer which resists disintegration in the stomach (such as an
enteric layer) and permits the inner component to pass intact into
the duodenum or a layer which delays or sustains release. A variety
of enteric and non-enteric layer or coating materials (such as
polymeric acids, shellacs, acetyl alcohol, and cellulose acetate or
combinations thereof) may be used.
[0235] Compounds of the invention may also be administered via a
slow release composition; wherein the composition includes a
compound of the invention and a biodegradable slow release carrier
(e.g., a polymeric carrier) or a pharmaceutically acceptable
non-biodegradable slow release carrier (e.g., an ion exchange
carrier).
[0236] Biodegradable and non-biodegradable slow release carriers
are well known in the art. Biodegradable carriers are used to form
particles or matrices which retain an active agent(s) and which
slowly degrade/dissolve in a suitable environment (e.g., aqueous,
acidic, basic and the like) to release the agent. Such particles
degrade/dissolve in body fluids to release the active compound(s)
therein. The particles are preferably nanoparticles or
nanoemulsions (e.g., in the range of about 1 to 500 nm in diameter,
preferably about 50-200 nm in diameter, and most preferably about
100 nm in diameter). In a process for preparing a slow release
composition, a slow release carrier and a compound of the invention
are first dissolved or dispersed in an organic solvent. The
resulting mixture is added into an aqueous solution containing an
optional surface-active agent(s) to produce an emulsion. The
organic solvent is then evaporated from the emulsion to provide a
colloidal suspension of particles containing the slow release
carrier and the compound of the invention.
[0237] The compound disclosed herein may be incorporated for
administration orally or by injection in a liquid form such as
aqueous solutions, suitably flavored syrups, aqueous or oil
suspensions, flavored emulsions with edible oils such as cottonseed
oil, sesame oil, coconut oil or peanut oil and the like, or in
elixirs or similar pharmaceutical vehicles. Suitable dispersing or
suspending agents for aqueous suspensions, include synthetic and
natural gums such as tragacanth, acacia, alginate, dextran, sodium
carboxymethylcellulose, methylcellulose, polyvinyl-pyrrolidone, and
gelatin. The liquid forms in suitably flavored suspending or
dispersing agents may also include synthetic and natural gums. For
parenteral administration, sterile suspensions and solutions are
desired. Isotonic preparations, which generally contain suitable
preservatives, are employed when intravenous administration is
desired.
[0238] The compounds may be administered parenterally via
injection. A parenteral formulation may consist of the active
ingredient dissolved in or mixed with an appropriate inert liquid
carrier. Acceptable liquid carriers usually comprise aqueous
solvents and other optional ingredients for aiding solubility or
preservation. Such aqueous solvents include sterile water, Ringer's
solution, or an isotonic aqueous saline solution. Other optional
ingredients include vegetable oils (such as peanut oil, cottonseed
oil, and sesame oil), and organic solvents (such as solketal,
glycerol, and formyl). A sterile, non-volatile oil may be employed
as a solvent or suspending agent. The parenteral formulation is
prepared by dissolving or suspending the active ingredient in the
liquid carrier whereby the final dosage unit contains from 0.005 to
10% by weight of the active ingredient. Other additives include
preservatives, isotonizers, solubilizers, stabilizers, and
pain-soothing agents. Injectable suspensions may also be prepared,
in which case appropriate liquid carriers, suspending agents and
the like may be employed.
[0239] Compounds of the invention may be administered intranasally
using a suitable intranasal vehicle.
[0240] In another embodiment, the compounds of this invention may
be administered directly to the lungs by inhalation.
[0241] Compounds of the invention may also be administered
topically or enhanced by using a suitable topical transdermal
vehicle or a transdermal patch.
[0242] For ocular administration, the composition is preferably in
the form of an ophthalmic composition. The ophthalmic compositions
are preferably formulated as eye-drop formulations and filled in
appropriate containers to facilitate administration to the eye, for
example a dropper fitted with a suitable pipette. Preferably, the
compositions are sterile and aqueous based, using purified water.
In addition to the compound of the invention, an ophthalmic
composition may contain one or more of:
[0243] a) a surfactant such as a polyoxyethylene fatty acid ester;
b) a thickening agents such as cellulose, cellulose derivatives,
carboxyvinyl polymers, polyvinyl polymers, and
polyvinylpyrrolidones, typically at a concentration n the range of
about 0.05 to about 5.0% (wt/vol); c) (as an alternative to or in
addition to storing the composition in a container containing
nitrogen and optionally including a free oxygen absorber such as
Fe), an anti-oxidant such as butylated hydroxyanisol, ascorbic
acid, sodium thiosulfate, or butylated hydroxytoluene at a
concentration of about 0.00005 to about 0.1% (wt/vol); d) ethanol
at a concentration of about 0.01 to 0.5% (wt/vol); and e) other
excipients such as an isotonic agent, buffer, preservative, and/or
pH-controlling agent. The pH of the ophthalmic composition is
desirably within the range of 4 to 8.
[0244] In certain embodiments, the composition of this invention
includes one or more additional agents. The other therapeutic agent
may be ay agent that is capable of treating, preventing or reducing
the symptoms of a tetracycline-responsive disease or disorder.
Alternatively, the other therapeutic agent may be any agent of
benefit to a patient when administered in combination with the
tetracycline compound in this invention.
[0245] While this invention has been particularly shown and
described with references to example embodiments thereof, it will
be understood by those skilled in the art that various changes in
form and details may be made therein without departing from the
scope of the invention encompassed by the appended claims.
EXEMPLIFICATION
[0246] The following abbreviations are used in throughout the
application.
[0247] Ac acetyl
[0248] AIBN 2,2'-azobis(2-methylpropionitrile)
[0249] aq aqueous
[0250] Bn benzyl
[0251] Boc tert-butoxycarbonyl
[0252] Bu butyl
[0253] Cb zbenzyloxycarbonyl
[0254] Cy tricyclohexylphosphine
[0255] dba dibenzylideneacetone
[0256] DIBAL H diisobutylaluminum hydride
[0257] DIEA N,N-diisopropylethylamine
[0258] DMAP 4-(dimethylamino)pyridine
[0259] DME 1,2-dimethoxyethane
[0260] DMF N,N-dimethylformamide
[0261] DMPU 1,3-dimethyl -3,4-5,6-tetrahydro-2(1H)-pyrimidone
[0262] DMSO dimethyl sulfoxide
[0263] EDC N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide
[0264] ESI electrospray ionization
[0265] E tethyl
[0266] EtOAc ethyl acetate
[0267] HPLC high performance liquid chromatography
[0268] HOBt 1-hydroxybenzotriazole
[0269] i iso
[0270] IBX 2-iodoxybenzoic acid
[0271] LDA lithium diisopropylamide
[0272] LHMDS lithium bis(trimethylsilyl)amide
[0273] LTMP lithium 2,2,6,6-tetramethylpiperidide
[0274] MeOH methanol
[0275] Ms methanesulfonyl
[0276] MS mass spectrometry
[0277] MTBE methyl tent-butyl ether
[0278] MW molecular weight
[0279] NBS N-bromosuccinimide
[0280] NCS N-chlorosuccinimide
[0281] NMR nuclear magnetic resonance spectrometry
[0282] Ph phenyl
[0283] Pr propyl
[0284] s secondary
[0285] t tertiary
[0286] TMEDA N,N,N'N'-tetramethylethylenediamine
[0287] TBS tert-butyldimethylsilyl
[0288] TEA triethylamine
[0289] Tf trifluoromathanesulfonyl
[0290] TFA trifluoroacetic acid
[0291] TFAA trifluoroacetic anhydride
[0292] THF tetrahydrofuran
[0293] TLC thin layer chromatography
[0294] Ts para-toluenesulfonyl
[0295] TsOH para-toluenesulfonic acid
[0296] Xantphos 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene
Example 1
[0297] Synthesis of Compound 100. Compound 100 was prepared
according to Scheme 1, below.
##STR00254##
[0298] 3,6-dimethoxy-2-methylbenzoic acid (1-2). nBuLi (8.6 mL,
13.7 mmol, 5.0 equiv) was added to a THF solution (5 mL) of
tetramethylpiperidine (2.3 mL, 13.7 mmol, 5.0 equiv) at 0.degree.
C. The reaction was stirred at 0.degree. C. for 30 min. To the
resulting solution was added a THF solution of 2,5-dimothoxybenzoic
acid (1-1, 500 mg, 2.75 mmol) at 0.degree. C. The reaction was
stirred at 0.degree. C. for 2.5 h. MeI (1.0 mL, 16.5 mmol, 6.0
equiv) was added to the reaction mixture dropwise. The reaction was
allowed to warm to 25.degree. C. over 1 h and stirred at 25.degree.
C. for 1 h. NaOH (6 N, 20 mL) was added. The resulting mixture was
extracted with t-butylmethyl ether (20 mL.times.2). The aqueous
layer was acidified with HCl (6 N) to pH 1 and extracted with EtOAc
(20 mL.times.4). The combined EtOAc extracts were dried
(Na.sub.2SO.sub.4) and concentrated to give 350 mg of crude product
1-2.
[0299] Phenyl 3,6-dimethoxy-2-methylbenzoate (1-3). Oxalyl chloride
(0.61 mL, 7.1 mmol, 4.0 equiv) was added to CH.sub.2Cl.sub.2
solution (15 mL, anhydrous) of crude 1-2 (350 mg, 1.79 mmol). DMF
(0.1 mL) was added to the resulting mixture. The reaction was
stirred at 25.degree. C. for 1 h and concentrated. The resulting
solid was re-dissolved in 15 mL of anhydrous CH.sub.2Cl.sub.2.
Phenol (337 mg, 3.58 mmol, 2.0 equiv), DMAP (437 mg, 3.58 mmol, 2.0
equiv), and triethylamine (1.20 mL, 8.95 mmol, 5.0 equiv) were
added to the reaction mixture. The reaction was stirred at
25.degree. C. for 12 h and concentrated. EtOAc and H.sub.2O were
added to the residue. The organic layer was washed with NaOH (1 N),
H2O, and brine, dried (Na.sub.2SO.sub.4), and concentrated. Flash
chromatography on silica gel (20:1 hexanes/EtOAc) yielded 291 mg of
compound 1-3 (39% for 2 steps).
[0300] Phenyl 6-hydroxy-3-methoxy-2-methylbenzoate (1-4). BBr.sub.3
(1.9 mL, 1.0 M, 1.9 mmol, 0.9 equiv) was added to a
CH.sub.2Cl.sub.2 solution (10 mL) of 1-3 (582 mg, 2.14 mmol) at
-78.degree. C. The reaction was stirred from -78.degree. C. to
25.degree. C. for 1.5 h, quenched with saturated NaHCO.sub.3 and
concentrated. EtOAc and H.sub.2O were added to the reaction
mixture. The aqueous layer was extracted with EtOAc. The combined
EtOAc extracts were dried (Na.sub.2SO.sub.4) and concentrated to
yield 480 mg of crude 1-4.
[0301] Phenyl 6-(tert-butoxycarbonyloxy)-3-methoxy-2-methylbenzoate
(1-5). Boc.sub.2O (487 mg, 2.23 mmol, 1.2 equiv) and DMAP (20 mg,
0.16 mmol, 0.1 equiv) were added to a CH.sub.2Cl.sub.2 solution of
crude 1-4 (480 mg). The reaction was stirred at 25.degree. C. for
1.5 h and concentrated. Flash chromatography on silica gel (15:1
hexanes/EtOAc) yielded 530 mg of compound 1-5 (80% for 2
steps).
[0302]
(4aS,11aR,12aS,13S)-3-(benzyloxy)-4a-(tert-butyldimethylsilyloxy)-1-
3-(dimethylamino)-5-hydroxy-10-methoxy-4,6-dioxo-4,4a,6,11,11a,12,12a,13-o-
ctahydrotetraceno[2,3-dfisoxazol-7-yl tert-butyl carbonate (1-7). A
THF solution (8 mL) of 1-5 (520 mg, 1.45 mmol, 2.5 equiv) was added
to a THF solution (8 mL) of LDA (6.50 mL, 10% wt, 4.36 mmol, 7.5
equiv) and TMEDA (1.0 mL, 7.3 mmol, 12.5 equiv) at -78.degree. C.
The reaction was stirred at -78.degree. C. for 5 min. A THF
solution (8 mL) of enone 1-6 (280 mg, 0.58 mmol, 1.0 equiv) was
added to the reaction mixture dropwise. The enone 1-6 was prepared
as described in PCT publication WO 2005/112945 and WO 2007/117639.
The reaction was stirred from -78.degree. C. to 25.degree. C. for 1
h, quenched with saturated NH.sub.4Cl, and extracted with EtOAc.
The combined EtOAc extracts were dried (Na.sub.2SO.sub.4) and
concentrated to yield the crude product. Preparative reverse phase
HPLC purification on a Waters Autopurification system using a
Sunfire Prep C18 OBD column [5 .mu.m, 19.times.50 mm; flow rate, 20
mL/min; Solvent A: H.sub.2O with 0.1% HCO.sub.2H; Solvent B:
CH.sub.3CN with 0.1% HCO.sub.2H; injection volume: 4.0 mL
(CH.sub.3CN); gradient: 80.fwdarw.100% B over 15 min; mass-directed
fraction collection]. Fractions with the desired MW were collected
and concentrated on a RotaVap at 25.degree. C. to remove most of
the acetonitrile. The resulting mostly aqueous solution was
extracted with EtOAc. The combined EtOAc extracts were dried
(Na.sub.2SO.sub.4) and concentrated to give 290 mg of pure 1-7
(67%).
[0303]
(4aS,11aR,12aS,13S)-3-(benzyloxy)-13-(dimethylamino)-4a,5,7-trihydr-
oxy-10-methoxy-11a,12,12a,13-tetrahydrotetraceno[2,3-dfisoxazole-4,6(4aH,1-
1H)-dione (1-8). Aqueous HF (2.4 mL, 48%) and TFA (0.1 mL) were
added to a CH.sub.3CN solution (9 mL) of 1-7 (210 mg, 0.29 mmol) in
a polypropylene tube at 25.degree. C. The reaction was stirred at
25.degree. C. for 18 h. The resulting mixture was poured into an
aqueous solution of K.sub.2HPO.sub.4 (21 g, dissolved in 150 mL
water). The mixture was extracted with EtOAc. The combined EtOAc
extracts were dried (Na.sub.2SO.sub.4) and concentrated to yield 86
mg of crude 1-8.
[0304] Compound 100. Palladium on carbon (10 mg, 10 wt %) was added
to a MeOH/dioxane solution (4 mL/4 mL) of crude 1-8 (86 mg). The
reaction was purged with hydrogen and stirred under H.sub.2
(balloon) at 25.degree. C. for 1 h. The reaction mixture was
filtered through a small Celite plug. The filtrate was concentrated
to yield the crude product. Preparative reverse phase HPLC
purification on a Waters Autopurification system using a Phenomenex
Polymerx.TM. 10.mu. RP-1 100A column [10 .mu.m, 150.times.21.20 mm;
flow rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B:
CH.sub.3CN; injection volume: 4.0 mL (0.05 N HCl/water); gradient:
0.fwdarw.100% B over 15 min; mass-directed fraction collection].
Fractions with the desired MW were collected and freeze-dried to
yield 47 mg of Compound 100 (81% for 2 steps): .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 7.21 (d, J=8.2 Hz, 1H), 6.78 (d, J=8.2 Hz,
1H), 4.05 (s, 1H), 3.78 (s, 3H), 3.08-2.90 (m, 3H), 3.01 (s, 3H),
2.94 (s, 3H), 2.20-2.11 (m, 2H), 1.67-1.56 (m, 1H); MS (ESI) m/z
445.23 (M+H).
Example 2
Synthesis of Compounds of Formula II, wherein Y is NH C(O) CH2
N(R2)(R3)
##STR00255##
[0306] Phenyl 6-(benzyloxy)-3-methoxy-2-methylbenzoate (2-1).
Phenol 1-4 (4.58 g, 17.7 mmol) was dissolved in anhydrous DMF (71
mL) and NaH (1.42 g, 35.5 g, 2 eq) was added. The mixture was
stirred at rt for 30 min. Benzylbromide (4.2 mL, 35.5 mmol, 2 eq)
was then added. After stirring for an overnight at rt, the mixture
was partitioned between EtOAc and H.sub.2O. The organic layer was
further washed with additional H.sub.2O three times and brine once.
The organic phase was dried over Na.sub.2SO.sub.4, filtered and
concentrated with rotavapor. Purification of the residue by flash
chromatography (silica gel, 97:3 hexanes/EtOAc) gave 2-1 (4.09 g)
as white solid:.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.45-7.22
(m, 8H), 7.13 (m, 2H), 6.82 (m, 2H), 5.11 (s, 2H), 3.81 (s, 3H),
2.32 (s, 3H); MS (ESI) m/z 371.2 (M+Na), calcd for
C.sub.22H.sub.20NaO.sub.4 371.14.
[0307] Phenyl 2-(benzyloxy)-5-methoxy-6-methyl-3-nitrobenzoate
(2-2). Solid Cu(NO.sub.3).sub.2.xH.sub.2O (3.006 g, 12.9 mmol, 1.1
eq) was added to a stirred solution of the compound 2-1 (4.09 g,
11.8 mmol) in acetic anhydride (47 mL) at 0.degree. C. The reaction
mixture was stirred at 0.degree. C. for 1 h and rt for 2 h, then
poured into 100 mL ice-water. Stirring was continued for another 1
h. The resulting yellow precipitate was collected by filtration.
Further purification with flash chromatography (silica gel, 95:5
hexanes/EtOAc) yielded compound 2-2 as light yellow solid (3.58 g):
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.50 (s, 1H), 7.47-7.25
(m, 8H), 7.02-7.00 (m, 2H), 5.11 (s, 2H), 3.92 (s, 3H), 2.36 (s,
3H); MS (ESI) m/z 392.2 (M-H), calcd for C.sub.22H.sub.18NO.sub.6
392.12.
[0308] Phenyl 2-(benzyloxy)-5-methoxy-6-methyl-3-aminobenzoate
(2-3). A mixture of nitro 2-2 (3.58 g, 9.11 mmol),
Na.sub.2S.sub.2O.sub.4 (9.33 g, 45.5 mmol, 5 equiv), 136 mL THF,
and 87 mL H.sub.2O was stirred at rt for an overnight. After
removing most of THF with rotavapor, the aqueous solution was
extracted with EtOAc three times. The combined organic layers were
washed with brine, dried over Na.sub.2SO.sub.4, filtered and
concentrated. The crude product 2-3 was used directly in the next
step.
[0309] Phenyl
2-(benzyloxy)-3-(bis(tert-butoxycarbonyl)amino)-5-methoxy-6-methylbenzoat-
e (2-4). Di-tert-butyl dicarbonate (5.04 g, 23.1 mmol, 2.5 eq) and
DMAP (56 mg, 0.46 mmol, 0.05 eq) were added to the solution of 2-3
(3.36 g, 9.24 mmol) in anhydrous DMF (92 mL). The resulting mixture
was stirred at rt for 5 h, and then diluted with EtOAc. The
solution was washed with H.sub.2O three times, brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated. Further purification
with flash chromatography (silica gel, 9:1 hexanes/EtOAc) yielded
compound 2-4 as white solid (4.20 g): .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.34-7.14 (m, 8H), 6.96 (d, J=7.8 Hz, 2H), 6.65
(s, 1H), 4.86 (s, 2H), 3.76 (s, 3H), 2.24 (s, 3H), 1.33 (s, 18H);
MS (ESI) m/z 586.2 (M+Na), calcd for C.sub.32H.sub.37NNaO.sub.8
586.25.
[0310] Intermediate 2-5.
##STR00256##
A solution of nBuLi in hexanes (1.60 M, 5.70 mL, 9.13 mmol, 1.1 eq)
was added dropwise to a solution of .sup.iPr.sub.2NH (1.29 mL, 9.13
mmol, 1.1 eq) and TMEDA (1.49 mL, 9.96 mmol, 1.2 eq) in THF (23 mL)
at -78.degree. C. The resulting solution was stirred at -78.degree.
C. for 1 h whereupon a solution of compound 2-4 (4.68 g, 8.30 mmol,
1 eq) in THF (30 mL) was added dropwise via cannula (colorless to
dark orange-red). After completion of addition, the mixture was
stirred for another 30 min at -78.degree. C., and then cooled to
-100.degree. C. Pre-cooled to -78.degree. C. solution of enone 1-6
(2.0 g, 4.15 mmol, 0.5 eq) in THF (30 mL) was added dropwise via
cannula. The resulting red color mixture was allowed to warm to
-20.degree. C. in 2 h. The reaction was quenched with sat. aq.
NH.sub.4Cl, then extracted with EtOAc three times. The combined
EtOAc extracts were washed with brine, dried (Na.sub.2SO.sub.4),
and concentrated. Purification of the residue by flash
chromatography (silica gel, 95:5 to 85:15 hexanes/EtOAc) gave
desired product 2-5 (3.589 g) as a light yellow foam: .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 16.05 (s, 1H), 7.52-7.26 (m, 10H),
6.85 (s, 1H), 5.35 (s, 2H), 4.97 (d, J=9.8 Hz, 1H), 4.73 (d, J=9.8
Hz, 1H), 3.99 (d, J=10.4 Hz, 1H), 3.80 (s, 3H), 3.36 (dd, J=16.5,
4.9 Hz, 1H), 2.98-2.92 (m, 1H), 2.60-2.40 (m, 8H), 2.36-2.28 (m,
1H), 2.16-2.13 (m, 1H), 1.38 (s, 18H), 0.80 (s, 9H), 0.24 (s, 3H),
0.13 (s, 3H); MS (ESI) m/z 952.59 (M+H), calcd for
C.sub.52H.sub.66N.sub.3O.sub.12Si 952.43.
[0311]
(4aS,11aR,12aS,13S)-8-amino-3,7-bis(benzyloxy)-4a-(tert-butyldimeth-
ylsilyloxy)-13-(dimethylamino)-5-hydroxy-10-methoxy-11a,12,12a,13-tetrahyd-
rotetraceno[2,3-d]isoxazole-4,6(4aH,11H)-dione (2-6). To a solution
of compound 2-5 (468 mg, 0.49 mmol) in anhydrous dioxane (9 mL) was
added solution of HCl in dioxane (4M, 9 mL) at rt. The resulting
mixture was stirred at rt and the reaction was monitored by LC-MS.
The volatiles were evaporated after SM was completely consumed. The
residue was suspended in EtOAc, and washed with sat. NaHCO.sub.3,
brine, dried (Na.sub.2SO.sub.4), and concentrated. Purification of
the residue by flash chromatography (silica gel, 80:20
hexanes/EtOAc) gave desired product 2-6 (422 mg) as a light yellow
foam: .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 16.14 (s, 1H),
7.52-7.26 (m, 10H), 6.48 (s, 1H), 5.35 (s, 2H), 4.88 (d, J=9.8 Hz,
1H), 4.81 (d, J=9.8 Hz, 1H), 3.84 (br s, 1H), 3.74 (s, 3H), 3.22
(dd, J=16.5, 4.9 Hz, 1H), 2.98-2.88 (m, 1H), 2.60-2.40 (m, 8H),
2.31-2.20 (m, 1H), 2.16-2.10 (m, 1H), 0.82 (s, 9H), 0.27 (s, 3H),
0.13 (s, 3H); MS (ESI) m/z 752.3 (M+H), calcd for
C.sub.42H.sub.50N.sub.3O.sub.8Si 752.33.
[0312] The choice of amine NHR.sup.2R.sup.3 used to convert 2-6 to
2-7 varied depending upon the desired final product.
[0313] Compound 101. 2-6 (29 mg, 0.039 mmol) was dissolved in THF
(1.5 mL).
[0314] bromoacetyl bromide (4.0 .mu.L, 0.046 mmol) was added. After
stirred at rt for 30 min, pyrrolidine (16.0 .mu.L, 0.19 mmol) was
added. Stirring was continued at rt. The reaction mixture was
poured into brine after SM was completely consumed and extracted
with EtOAc three times. The combined EtOAc layers were dried with
Na.sub.2SO.sub.4 and concentrated to give a crude 2-7 wherein
R.sup.2 and R.sup.3 are taken together to from pyrrolidine, which
was used directly for the next step without purification. In a
plastic vial, 2-7 was dissolved in CH.sub.3CN (1 mL). Aqueous HF
(48%, 0.25 mL) was added. After stirred at rt for 16 h, the
reaction mixture was poured into aqueous solution (12.5 mL) of
K.sub.2HPO.sub.4 (1.75 g). The resulting mixture was extracted
three times with EtOAc. The combined organic phases were washed
with brine, dried, concentrated to give crude product. The above
crude was dissolved in 0.5 N HCl in MeOH (155 .mu.L, 2 eq). The
excess volatiles were evaporated. The pre-formed HCl salt was
re-dissolved in MeOH (2.0 mL) and to the resulting solution was
added palladium on carbon (10% wt, 9.0 mg, 30% w/w). The reaction
flask was briefly evacuated and re-filled with hydrogen. The
reaction mixture was stirred at rt and monitored by LC-MS. After SM
was consumed, the mixture was filtered through a small pad of
Celite. The filtrate was concentrated to give crude, which was
purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HC1/water; Solvent B: CH3CN; injection volume: 4.0 mL (0.05
N HCl/water); gradient: 10.fwdarw.50% B over 10 min; mass-directed
fraction collection]. Fractions with the desired MW, eluting at
5.14-5.60 min, were collected and freeze-dried to give Compound 101
as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.11
(s, 1H), 4.32 (s, 2H), 4.09 (s, 1H), 3.81-3.75 (m, 5H), 3.30-3.15
(m, 3H), 3.10-2.90 (m, 8H), 2.25-2.00 (m, 6H), 1.68-1.54 (m, 1H);
MS (ESI) m/z 571.2 (M+H), calcd for C.sub.28H.sub.35N.sub.4O.sub.9
571.23.
[0315] Compound 102. Compound 102 was obtained by the procedure of
Compound 101 employing azetidine as NR.sup.2R.sup.3. Crude product
was purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu., RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 10.fwdarw.40% B over 10 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 5.36-5.80 min, were collected and freeze-dried to give
Compound 102 as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.08 (s, 1H), 4.42-4.34 (m, 4H), 4.25-4.18 (m, 2H), 4.10
(s, 1H), 3.77 (s, 3H), 3.28-3.20 (m, 1H), 3.08-2.92 (m, 8H),
2.70-2.62 (m, 1H), 2.56-2.44 (m, 1H), 2.22-2.06 (m, 2H), 1.65-1.56
(m, 1H); MS (ESI) m/z 557.2 (M+H), calcd for
C.sub.27H.sub.33N.sub.4O.sub.9 557.22.
[0316] Compound 103. Compound 103 was obtained by the procedure of
Compound 101 employing piperidine as NR.sup.2R.sup.3. Crude product
was purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu., RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 10.fwdarw.30% B over 10 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 7.12-7.80 min, were collected and freeze-dried to give
Compound 103 as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.11 (s, 1H), 4.19 (s, 2H), 4.09 (s, 1H), 3.79 (s, 3H),
3.65-3.62 (m, 2H), 3.28-3.20 (m, 1H), 3.18-2.90 (m, 10H), 2.25-2.08
(m, 2H), 2.00-1.80 (m, 6H), 1.65-1.56 (m, 1H); MS (ESI) m/z 585.2
(M+H), calcd for C.sub.29H.sub.37N.sub.4O.sub.9 585.25.
[0317] Compound 104. Compound 104 was obtained by the procedure of
Compound 101 employing N-ethyl methylamine as NR.sup.2R.sup.3.
Crude product was purified by HPLC on a Waters Autopurification
system using a Phenomenex Polymerx.TM. 10.mu., RP-.gamma. 100 R
column [30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent
A: 0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0
mL (0.05 N HCl/water); gradient: 10.fwdarw.30% B over 10 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 5.80-6.30 min, were collected and freeze-dried to give
Compound 104 as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.12 (s, 1H), 4.30 (d, J=16.0 Hz, 1H), 4.19 (d, J=16.0 Hz,
1H), 4.10 (s, 1H), 3.80 (s, 3H), 3.45-3.20 (m, 3H), 3.10-2.90 (m,
11H), 2.25-2.08 (m, 2H), 1.65-1.56 (m, 1H), 1.40 (t, J=7.3 Hz, 3H);
MS (ESI) m/z 559.2 (M+H), calcd for C.sub.27H.sub.35N.sub.4O.sub.9
559.23.
[0318] Compound 105. Compound 105 was obtained by the procedure of
Compound 101 employing dimethylamine as NR.sup.2R.sup.3. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH3CN; injection volume: 4.0 mL (0.05
N HCl/water); gradient: 10.fwdarw.30% B over 10 min; mass-directed
fraction collection]. Fractions with the desired MW, eluting at
5.76-6.22 min, were collected and freeze-dried to give Compound 105
as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.12
(s, 1H), 4.24 (s, 2H), 4.09 (s, 1H), 3.80 (s, 3H), 3.30-3.25 (m,
1H), 3.10-2.90 (m, 14 H), 2.25-2.08 (m, 2H), 1.65-1.56 (m, 1H); MS
(ESI) m/z 545.2 (M+H), calcd for C.sub.26H.sub.33N.sub.4O.sub.9
545.22.
[0319] Compound 106. Compound 106 was obtained by the procedure of
Compound 101 employing isobutylamine as NR.sup.2R.sup.3. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 10.fwdarw.30% B over 10 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 7.36-8.05 min, were collected and freeze-dried to give
Compound 106 as a yellow solid: iH NMR (400 MHz, CD.sub.3OD)
.delta. 8.13 (s, 1H), 4.09 (s, 3H), 3.79 (s, 3H), 3.30-3.25 (m,
1H), 3.10-2.90 (m, 10H), 2.25-2.05 (m, 3H), 1.65-1.56 (m, 1H), 1.07
(d, J=6.9 Hz, 6H); MS (ESI) m/z 573.3 (M+H), calcd for
C.sub.28H.sub.37N.sub.4O.sub.9 573.25.
[0320] Compound 107. Compound 107 was obtained by the procedure of
Compound 101 employing isopropylamine as NR.sup.2R.sup.3. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu., RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 10.fwdarw.30% B over 10 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 6.10-6.54 min, were collected and freeze-dried to give
Compound 107 as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.13 (s, 1H), 4.09 (s, 3H), 3.79 (s, 3H), 3.45-3.55 (m,
1H), 3.30-3.25 (m, 1H), 3.10-2.90 (m, 8H), 2.25-2.10 (m, 2H),
1.65-1.56 (m, 1H), 1.38 (d, J=6.4 Hz, 6H); MS (ESI) m/z 559.3
(M+H), calcd for C.sub.27H.sub.35N.sub.4O.sub.9 559.23.
[0321] Compound 108. Compound 108 was obtained by the procedure of
Compound 101 employing cyclopropylamine as NR.sup.2R.sup.3. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 10.fwdarw.30% B over 10 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 5.82-6.50 min, were collected and freeze-dried to give
Compound 108 as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.13 (s, 1H), 4.09 (s, 3H), 3.79 (s, 3H), 3.45-3.55 (m,
1H), 3.30-3.25 (m, 1H), 3.10-2.90 (m, 8H), 2.25-2.10 (m, 2H),
1.65-1.56 (m, 1H), 1.38 (d, J=6.4 Hz, 6H); MS (ESI) m/z 557.3
(M+H), calcd for C.sub.27H.sub.33N.sub.4O.sub.9 557.22.
[0322] Compound 109. Compound 109 was obtained by the procedure of
Compound 101 employing tert-butylamine as NR.sup.2R.sup.3. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 10.fwdarw.30% B over 10 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 6.46-6.92 min, were collected and freeze-dried to give
Compound 109 as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.14 (s, 1H), 4.08 (s, 3H), 3.78 (s, 3H), 3.30-3.25 (m,
1H), 3.10-2.90 (m, 8H), 2.25-2.10 (m, 2H), 1.65-1.56 (m, 1H), 1.42
(s, 9H); MS (ESI) m/z 573.2 (M+H), calcd for
C.sub.28H.sub.37N.sub.4O.sub.9 573.25.
[0323] Compound 110. Compound 110 was obtained from 2-5 by HF
treatment and hydrogenation according to procedures used in the
preparation of Compound 101. Crude product was purified by HPLC on
a Waters Autopurification system using a Phenomenex Polymerx.TM.
10.mu. RP-.gamma. 100 R column [30.times.21.20 mm, 10 micron; flow
rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B:
CH.sub.3CN; injection volume: 4.0 mL (0.05 N HCl/water); gradient:
10.fwdarw.30% B over 10 min; mass-directed fraction collection].
Fractions with the desired MW, eluting at 4.88-5.78 min, were
collected and freeze-dried to give Compound 110 as a yellow solid:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.27 (s, 1H), 4.09 (s,
1H), 3.83 (s, 3H), 3.30-3.25 (m, 1H), 3.10-2.90 (m, 8H), 2.25-2.10
(m, 2H), 1.65-1.56 (m, 1H); MS (ESI) m/z 460.2 (M+H), calcd for
C.sub.22H.sub.26N.sub.3O.sub.8 460.16.
[0324] Compound 111. Compound 111 was obtained by the procedure of
Compound 101, substituting bromoacetyl bromide and pyrrolidine with
3,3,-dimethylbutyryl chloride. Crude product was purified by HPLC
on a Waters Autopurification system using a Phenomenex Polymerx.TM.
10.mu. RP-.gamma. 100 R column [30.times.21.20 mm, 10 micron; flow
rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B:
CH.sub.3CN; injection volume: 4.0 mL (0.05 N HCl/water); gradient:
10.fwdarw.30% B over 10 min; mass-directed fraction collection].
Fractions with the desired MW, eluting at 12.40-13.97 min, were
collected and freeze-dried to give Compound 111 as a yellow solid:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.99 (s, 1H), 4.07 (s,
1H), 3.79 (s, 3H), 3.30-3.25 (m, 1H), 3.10-2.90 (m, 8H), 2.34 (s,
2H), 2.25-2.10 (m, 2H), 1.65-1.56 (m, 1H), 1.10 (s, 9H); MS (ESI)
m/z 558.0 (M+H), calcd for C.sub.28H.sub.36N.sub.3O.sub.9
558.24.
[0325] Compound 112. Compound 112 was obtained by the procedure of
Compound 101 employing 3-fluoroazetidine as NR.sup.2R.sup.3. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N TFA/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 10.fwdarw.30% B over 20 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 12.35-13.50 min, were collected and freeze-dried to give
Compound 112 as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.10 (s, 1H), 5.51-5.37 (m, 1H), 4.70-4.39 (m, 6H), 4.05
(s, 1H), 3.79 (s, 3H), 3.30-3.25 (m, 1H), 3.10-2.90 (m, 8H),
2.25-2.10 (m, 2H), 1.65-1.56 (m, 1H); MS (ESI) m/z 575.2 (M+H),
calcd for C.sub.27H.sub.32FN.sub.4O.sub.9 575.21.
[0326] Compound 113. Compound 113 was obtained by the procedure of
Compound 101, substituting bromoacetyl bromide and pyrrolidine with
4,4,-dimethylpentyryl chloride. Crude product was purified by HPLC
on a Waters Autopurification system using a Phenomenex Polymerx.TM.
10.mu. RP-.gamma. 100 R column [30.times.21.20 mm, 10 micron; flow
rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B:
CH.sub.3CN; injection volume: 4.0 mL (0.05 N HCl/water); gradient:
15.fwdarw.60% B over 10 min; mass-directed fraction collection].
Fractions with the desired MW, eluting at 9.06-9.74 min, were
collected and freeze-dried to give Compound 113 as a yellow solid:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.02 (s, 1H), 4.07 (s,
1H), 3.78 (s, 3H), 3.30-3.25 (m, 1H), 3.10-2.90 (m, 8H), 2.48-2.44
(m, 2H), 2.25-2.10 (m, 2H), 1.65-1.56 (m, 3H), 0.96 (s, 9H); MS
(ESI) m/z 572.4 (M+H), calcd for C.sub.29H.sub.38N.sub.3O.sub.9
572.25.
[0327] Compound 114. Compound 114 was obtained by the procedure of
Compound 101 employing (R)-3-fluoropyrrolidine as NR.sup.2R.sup.3.
Crude product was purified by HPLC on a Waters Autopurification
system using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R
column [30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent
A: 0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0
mL (0.05 N HCl/water); gradient: 5.fwdarw.25% B over 12 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 9.05-11.05 min, were collected and freeze-dried to give
Compound 114 as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.13 (s, 1H), 5.55-5.42 (m, 1H), 4.45-4.41 (m, 2H),
4.15-3.95 (m, 3H), 3.79 (s, 3H), 3.50-3.40 (m, 2H), 3.30-3.25 (m,
1H), 3.10-2.90 (m, 8H), 2.50-2.35 (m, 2H), 2.25-2.10 (m, 2H),
1.65-1.56 (m, 1H); MS (ESI) m/z 589.3 (M+H), calcd for
C.sub.29H.sub.34FN.sub.4O.sub.9 589.22.
[0328] Compound 115. Compound 115 was obtained by the procedure of
Compound 101 employing (S)-3-fluoropyrrolidine as NR.sup.2R.sup.3.
Crude product was purified by HPLC on a Waters Autopurification
system using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R
column [30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent
A: 0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0
mL (0.05 N HCl/water); gradient: 5.fwdarw.30% B over 12 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 8.05-9.85 min, were collected and freeze-dried to give
Compound 115 as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.12 (s, 1H), 5.55-5.42 (m, 1H), 4.46-4.42 (m, 2H),
4.15-3.95 (m, 3H), 3.79 (s, 3H), 3.50-3.40 (m, 2H), 3.30-3.25 (m,
1H), 3.10-2.90 (m, 8H), 2.55-2.40 (m, 2H), 2.25-2.10 (m, 2H),
1.65-1.56 (m, 1H); MS (ESI) m/z 589.3 (M+H), calcd for
C.sub.29H.sub.34FN.sub.4O.sub.9 589.22.
[0329] Compound 116. Compound 116 was obtained by the procedure of
Compound 101 employing O-tert-butyl hydroxylamine as
NR.sup.2R.sup.3. .sup.1HNMR (400 MHz, CD.sub.3OD) .delta. 8.12 (s,
1H), 4.34 (s, 2H), 4.15 (s, 1H), 3.80 (s, 3H), 2.85-3.50 (m, 3H),
3.07 (s, 3H), 2.99 (s, 3H), 2.22-2.28 (m, 1H), 2.07-2.16 (m, 1H),
1.55-1.65 (m, 1H), 1.47 (s, 9H); MS (ESI) m/z 589.1 (M+H), calcd
for C.sub.28H.sub.37N.sub.4O.sub.10 589.25.
[0330] Compound 117. Compound 117 was obtained by the procedure of
Compound 101 substituting bromoacetyl bromide and an amine with
tert-butoxyacetyl chloride. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.31 (s, 1H), 4.08 (s, 1H), 4.07 (s, 2H), 3.80 (s, 3H),
3.24 (dd, J=7.3, 16.0 Hz, 1H), 2.90-3.10 (m, 2H), 3.05 (s, 3H),
2.97 (s, 3H), 2.15-2.22 (m, 1H), 2.05-2.21 (m, 1H), 1.55-1.65 (m,
1H), 1.32 (s, 9H); MS (ESI) m/z 574.2 (M+H), calcd for
C.sub.28H.sub.37N.sub.3O.sub.10 574.24.
[0331] Compound 118. Compound 118 was prepared similarly to
Compound 101 using ethylisopropylamine as NR.sup.2R.sup.3.
.sup.1HNMR (400 MHz, CD.sub.3OD) .delta. 8.13 (s, 1H), 4.32 (s,
J=17.5 Hz, 1H), 4.08 (s, 1H), 4.06 (d, J=17.5 Hz, 1H), 3.78 (s,
3H), 3.75-3.85 (m, 2H), 2.80-3.50 (m, 10H), 2.10-2.22 (m, 2H),
1.50-1.70 (m, 1H), 1.35-1.45 (m, 9H); MS (ESI) m/z 587.1 (M+H),
calcd for C.sub.29H.sub.39N.sub.4O.sub.9 587.27.
[0332] Compound 119. Compound 119 was prepared similarly to
Compound 101 using imidazole as NR.sup.2R.sup.3. .sup.1HNMR (400
MHz, CD.sub.3OD) .delta. 9.02 (s, 1H), 8.07 (s, 1H), 7.68 (s, 1H),
7.63 (s, 1H), 5.34 (s, 2H), 4.07 (s, 1H), 3.76 (s, 3H), 2.80-3.50
(m, 9H), 2.10-2.22 (m, 2H), 1.55-1.70 (m, 1H); MS (ESI) m/z 568.1
(M+H), calcd for C.sub.27H.sub.30N.sub.5O.sub.9 568.21.
Example 3
[0333] Synthesis of Certain Compounds of Formula II, wherein Y is
NH C(O) CH2 NH(R3), or NH C(O) heterocyclyl. Scheme 3 depicts the
synthesis of compounds of Formula II, wherein Y is NH C(O)
heterocyclyl or NH C(O) CH2 NH(R3) where R3 is not hydrogen.
##STR00257##
[0334] Compound 120. Compound 110 (40 mg crude, .about.0.08 mmol, 1
equiv) was dissolved in CH.sub.3CN and 3 drops of DMPU. Then
bromoacetyl bromide (24 mg, 0.12 mmol, 1.5 equiv) and
Na.sub.2CO.sub.3 (51 mg, 0.48 mmol, 6.0 equiv) were added into the
reaction mixture and stirred at rt for 1 hr. Ethylamine (0.80 mmol,
10.0 equiv) was added into the reaction mixture and stirred for 1
h. HCl/MeOH (, 1 mL 4 N) was added at 0.degree. C. and the solution
was stirred for 5 min, concentrated in vacuo, and purified by
reverse phase preparative HPLC to yield the desired Compound 120 as
a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.11 (s
1H), 4.10-4.09 (m, 3H), 3.79 (s, 3H), 3.23-2.95 (m, 11H), 2.25-2.10
(m, 2H), 1.69-1.53 (m, 1H), 1.37 (t, J=7.2 Hz, 3H); MS (ESI) m/z
545.2 (M+H).
[0335] Compound 121. Compound 121 was prepared similarly to
Compound 120, substituting ethylamine with 2-fluoroethylamine:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.08 (s 1H), 4.68 (t,
J=4.4 Hz, 1H), 4.11 (s, 2H), 4.01 (s, 1H), 3.74 (s, 3H), 3.49 (t,
J=4.8 Hz, 1H), 3.42 (t, J=4.8 Hz, 1H), 3.25-2.87 (m, 10H),
2.15-2.03 (m, 2H), 1.61-1.49 (m, 1H); MS (ESI) m/z 563.2 (M+H).
[0336] Compound 122. Compound 122 was prepared similarly to
Compound 120, substituting ethylamine with 2,2-difluoroethylamine:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.15 (s 1H), 6.36 (t,
J=44 Hz, 1H), 4.21 (s, 2H), 4.09 (s, 1H), 3.80 (s, 3H), 3.73-3.65
(m, 2H), 3.13-2.95 (m, 9H), 2.23-2.09 (m, 2H), 1.69-1.57 (m, 1H);
MS (ESI) m/z 581.1 (M+H).
[0337] Compound 123. Compound 123 was prepared similarly to
Compound 120, substituting ethylamine with n-propylamine: .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 8.14 (s 1H), 4.09 (s, 3H), 3.81
(s, 3H), 3.30-3.25 (m, 2H), 3.10-2.95 (m, 9H), 2.23-2.08 (m, 2H),
1.83-1.73 (m, 2H), 1.68-1.57 (m, 1H), 1.08 (t, J=7.2 Hz, 3H); MS
(ESI) m/z 559.2 (M+H).
[0338] Compound 124. Compound 124 was prepared similarly to
Compound 120, substituting ethylamine with 2-methoxyethylamine:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.13 (s, 1H), 4.13 (s,
2H), 4.05-3.93 (m, 1H), 3.80 (s, 3H), 3.75-3.65 (m, 2H), 3.44 (s,
3H), 3.36-3.23 (m, 3H), 3.10-2.90 (m, 8H), 2.25-2.05 (m, 2H),
1.68-1.59 (m, 1H); MS (ESI) m/z 575.2 (M+H).
[0339] Compound 125. Compound 125 was prepared similarly to
Compound 120, substituting ethylamine with n-hexylamine: .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 8.12 (s 1H), 4.06 (s, 3H), 3.80
(s, 3H), 3.12-2.91 (m, 11H), 2.19-2.08 (m, 2H), 1.75-1.55 (m, 3H),
1.45-1.32 (m, 6H), 0.92 (t, J=7.2 Hz, 3H); MS (ESI) m/z 601.3
(M+H).
[0340] Compound 126. Compound 126 was prepared similarly to
Compound 120, substituting ethylamine with cyclopropylmethylamine:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.13 (s 1H), 4.13 (s,
2H), 4.10 (s, 1H), 3.80 (s, 3H), 3.05-2.91 (m, 11H), 2.25-2.10 (m,
2H), 1.68-1.56 (m, 1H), 1.19-1.10 (m, 1H), 0.78-0.73 (m, 2H),
0.48-0.43 (m, 2H); MS (ESI) m/z 571.2 (M+H).
[0341] Compound 127. Compound 127 was prepared similarly to
Compound 120, substituting ethylamine with cyclobutylmethylamine:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33 (s 1H), 4.28 (s,
1H), 4.25 (s, 2H), 3.99 (s, 3H), 3.37-3.14 (m, 11H), 2.97-2.86 (m,
1H), 2.45-2.06 (m, 8H), 1.87-1.75 (m, 1H); MS (ESI) m/z 585.3
(M+H).
[0342] Compound 128. Compound 128 was prepared similarly to
Compound 120, substituting ethylamine with cyclopentylmethylamine
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.34 (s 1H), 4.29 (s,
3H), 4.07 (s, 3H), 3.30-3.10 (m, 11H), 2.50-2.30 (m, 3H), 2.18-2.10
(m, 2H), 1.95-1.78 (m, 4H), 1.55-1.46 (m, 3H); MS (ESI) m/z 599.3
(M+H).
[0343] Compound 129. Compound 129 was prepared similarly to
Compound 120, substituting ethylamine with cyclohexylmethylamine:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.14 (s 1H), 4.09 (s,
3H), 3.80 (s, 3H), 3.05-2.95 (m, 11H), 2.23-2.08 (m, 2H), 1.88-1.55
(m, 8H), 1.40-1.21 (m, 4H); MS (ESI) m/z 613.3 (M+H).
[0344] Compound 130. Compound 130 was prepared similarly to
Compound 120, substituting ethylamine with neopentylamine: .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 8.13 (s 1H), 4.11-4.10 (m, 3H),
3.80 (s, 3H), 3.26-3.20 (m, 1H), 3.08-2.95 (m, 10H), 2.25-2.06 (m,
2H), 1.66-1.54 (m, 1H), 1.12 (s, 9H); MS (ESI) m/z 587.3 (M+H).
[0345] Compound 131. Compound 131 was prepared similarly to
Compound 120, substituting ethylamine with cyclobutylamine: .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 8.11 (s 1H), 4.07 (s, 1H), 3.95
(s, 2H), 3.88-3.80 (m, 1H), 3.77 (s, 3H), 3.25-3.20 (m, 1H),
3.06-2.92 (m, 8H), 2.38-2.06 (m, 6H), 1.95-1.84 (m, 2H), 1.63-1.54
(m, 1H); MS (ESI) m/z 571.2 (M+H).
[0346] Compound 132. Compound 132 was prepared similarly to
Compound 120, substituting ethylamine with cyclopentylamine:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.15 (s 1H), 4.10 (s,
3H), 3.84 (s, 3H), 3.68-3.64 (m, 1H), 3.26-3.22 (m, 1H), 3.08-2.95
(m, 8H), 2.25-2.10 (m, 4H), 1.92-1.60 (m, 7H); MS (ESI) m/z 585.3
(M+H).
[0347] Compound 133. Compound 133 was prepared similarly to
Compound 120, substituting ethylamine with cyclohexylamine: .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 8.12 (s 1H), 4.08 (s, 2H), 4.07
(s, 1H), 3.86 (s, 3H), 3.22-2.90 (m, 10H), 2.22-2.08 (m, 4H),
1.95-1.85 (m, 2H), 1.68-1.55 (m, 2H), 1.46-1.30 (m, 5H); MS (ESI)
m/z 599.3 (M+H).
[0348] Compound 134. Compound 134 was prepared similarly to
Compound 120, substituting ethylamine with aniline: .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 8.22 (s 1H), 7.56-7.30 (m, 5H), 4.34
(s, 2H), 4.09 (s, 1H), 3.86 (s, 3H), 3.14-2.95 (m, 9H), 2.25-2.10
(m, 2H), 1.68-1.58 (m, 1H); MS (ESI) m/z 593.2 (M+H).
[0349] Compound 135. Compound 135 was prepared similarly to
Compound 120, substituting ethylamine with N-methylpropylamine:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.11 (s 1H), 4.32 (d,
J=16.0 Hz, 1H), 4.20 (d, J=16.0 Hz, 1H), 4.10 (s, 1H), 3.80 (s,
3H), 3.27-2.90 (m, 14H), 2.25-2.10 (m, 2H), 1.88-1.79 (m, 2H),
1.67-1.57 (m, 1H), 1.05 (t, J=7.2 Hz, 3H); MS (ESI) m/z 573.3
(M+H).
[0350] Compound 136. Compound 136 was prepared similarly to
Compound 120, substituting ethylamine with diethylamine: .sup.1H
NMR (400 MHz, CD.sub.3OD) .delta. 8.10 (s 1H), 4.22 (s, 2H), 4.05
(s, 1H), 3.79 (s, 3H), 3.46-3.22 (m, 5H), 3.07-2.90 (m, 8H),
2.20-2.07 (m, 2H), 1.66-1.57 (m, 1H), 1.38 (t, J=7.2 Hz, 6H); MS
(ESI) m/z 573.2 (M+H).
[0351] Compound 137. Compound 137 was prepared similarly to
Compound 120, substituting ethylamine with
3-azabicyclo[3,3,0]octane: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.07 (d, J=9.2 Hz, 1H), 4.25-4.16 (m, 2H), 4.15-4.05 (m,
1H), 4.02 (s, 1H), 3.95-3.80 (m, 1H), 3.74 (s, 3H), 3.63-3.54 (m,
1H), 3.43-3.35 (m, 1H), 3.07-2.85 (m, 9H), 2.75-2.60 (m, 2H),
2.15-2.02 (m, 2H), 1.65-1.50 (m, 7H); MS (ESI) m/z 611.3 (M+H).
[0352] Compound 138. Compound 138 was prepared similarly to
Compound 120, substituting ethylamine with isoindoline: .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 8.16 (s 1H), 7.42 (s, 4H), 5.10-5.00
(m, 2H), 4.73-4.62 (m, 2H), 4.55 (s, 2H), 4.07 (s, 1H), 3.79 (s,
3H), 3.25-3.17 (m, 1H), 3.07-2.90 (m, 8H), 2.22-2.08 (m, 2H),
1.67-1.55 (m, 1H); MS (ESI) m/z 619.2 (M+H).
[0353] Compound 139. Compound 139 was prepared similarly to
Compound 120, substituting bromoacetyl bromide/amine with
3-dimethylaminobenzoyl chloride: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.16 (s 1H), 8.02 (d, J=8.0 Hz, 1H), 7.91 (s, 1H),
7.85-7.82 (m, 1H), 7.71-7.67 (m, 1H), 4.01 (s, 1H), 3.73 (s, 3H),
3.28 (s, 6H), 3.20-3.16 (m, 1H), 3.01-2.85 (m, 8H), 2.20-1.98 (m,
2H), 1.57-1.48 (m, 1H), MS (ESI) m/z 607.1 (M+H).
[0354] Compound 140. Compound 140 was prepared similarly to
Compound 120, substituting bromoacetyl bromide/amine with nicotinyl
chloride: .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.41 (s 1H),
9.14 (d, J=8.4 Hz, 1H), 9.06 (d, J=5.6 Hz, 1H), 8.23 (d, J=2.0 Hz,
1H), 8.04 (s, 1H), 4.01 (s, 1H), 3.84 (s, 3H), 3.20-3.16 (m, 1H),
3.08-2.95 (m, 8H), 2.25-2.12 (m, 2H); 1.70-1.58 (m, 1H), MS (ESI)
m/z 565.1 (M+H).
Example 4
[0355] Synthesis of Certain Compounds of Formula II, wherein Y is
--NH--C(O)-(saturated heterocyclyl). Scheme 4 depicts the synthesis
of other compounds of Formula II, wherein Y is
--NH--C(O)-(saturated heterocyclyl).
##STR00258##
[0356] In Scheme 4, "PG" represents a protecting group and R.sup.A
represents hydrogen, (C.sub.1-C.sub.5)alkyl, --(C.sub.0-C.sub.5)
alkylene-carbocyclyl, or --(C.sub.0-C.sub.5)alkylene-heterocyclyl.
For all of the compounds made by Scheme 4 and described below,
R.sup.Z is hydrogen and R.sup.X and R.sup.Y are taken together with
the carbon and nitrogen atoms to which they are respectively bound
to form an optionally substituted 4-7 membered saturated
heterocyclyl. It will be readily apparent to those of skill in the
art, however, that this Scheme 4 will also be useful to synthesize
compounds where R.sup.X, R.sup.Y and R.sup.Z are R.sup.2, R.sup.5b
and R.sup.5b, respectively, as defined in structural formula
(I).
[0357] Compound 141.
##STR00259##
[0358] A solution of (S)-(-)-N-(trifluoroacetyl)prolyl chloride in
DCM (0.1 M, 383 .mu.L, 0.038 mmol, 1.2 equiv) was added to a
solution of aniline 2-6 (24 mg, 0.032 mmol, 1.0 equiv) in THF (2
mL). The resulting light orange solution was stirred at rt for 5
min, diluted with brine (10 mL). The resulting mixture was
extracted with EtOAc (2.times.15 mL). The combined organic extracts
were dried (Na.sub.2SO.sub.4), filtered and concentrated to yield
the crude product 4-1-1: MS (ESI) m/z 945.38 (M+H). Aqueous HF
(48-50%, 0.2 mL) was added to a solution of the above crude product
4-1-1 in acetonitrile (0.5 mL) in a polypropylene reaction vessel
at 23.degree. C. The resulting mixture was stirred vigorously at
23.degree. C. overnight and poured into aqueous K.sub.2HPO.sub.4
(2.5 g dissolved in 20 mL water). The resulting mixture was
extracted with EtOAc (30 mL, then 2.times.10 mL). The combined
organic extracts were dried over anhydrous sodium sulfate,
filtered, and concentrated. The residue was used directly in the
next step without further purification.
[0359] The above crude product was dissolved in MeOH (2 mL) and
dioxane (1 mL). Pd--C (10 wt %, 6 mg) was added in one portion at
23.degree. C. The reaction vessel was sealed and purged with
hydrogen by briefly evacuating the flask followed by flushing with
hydrogen gas (1 atm). After stirring at 23.degree. C. for 4 hrs,
more Pd--C (10 wt %, 5 mg) was added. The resulting mixture was
stirred for 2 h and filtered through a small Celite pad. The
filtrate was concentrated. The residue was purified by a
preparative reverse phase HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100A column [10
.mu.m, 150.times.21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05 N
HCl/water; Solvent B: CH.sub.3CN; injection volume: 3.0 mL (0.05 N
HCl/water and MeCN, 1:1); gradient: 15.fwdarw.60% B over 10 min;
mass-directed fraction collection]. Fractions containing the
desired product, eluting at 7.3-8.0 min, were collected and
freeze-dried to yield Compound 141 (11.9 mg). The product was
re-purified by a preparative reverse phase HPLC on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu.
RP-.gamma. 100A column [10 .mu.m, 150.times.21.20 mm; flow rate, 20
mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH.sub.3CN;
injection volume: 3.0 mL (0.05 N HCl/water and MeCN, 1:1);
gradient: 15.fwdarw.60% B over 20 min; mass-directed fraction
collection]. Fractions containing the desired product, eluting at
11.4-11.8 min, were collected and freeze-dried to yield Compound
141 (8.4 mg, 38% over 3 steps): .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.03 (s, 1H), 4.81-4.78 (m, 1H), 4.07 (s, 1H), 3.85 (t,
J=6.4 Hz, 2H), 3.78 (s, 3H), 3.27-3.21 (m, 1H), 3.03-2.92 (m, 8H),
2.40-2.32 (m, 1H), 2.20-2.03 (m, 5H), 1.66-1.56 (m, 1H); MS (ESI)
m/z 653.31 (M+H).
[0360] Compound 142.
##STR00260##
[0361] K.sub.2CO.sub.3 (14.7 mg, 0.106 mmol, 1.6 equiv) was added
in one portion to a solution of crude product 4-1-1 (0.067 mmol,
1.0 equiv) in a mixture of MeOH (2 mL), THF (0.5 mL) and water (0.3
mL). The resulting orange reaction mixture was stirred at rt
overnight. More K.sub.2CO.sub.3 (9 mg, 0.065 mmol, 0.98 equiv), and
the reaction mixture was stirred at rt overnight. The reaction
mixture was then diluted with brine (20 mL) and extracted with
EtOAc (50 mL, then 20 mL). The combined organic extracts were dried
over anhydrous sodium sulfate, filtered, and concentrated. The
residue was purified by a Preparative reverse phase HPLC on a
Waters Autopurification system using a Sunfire Prep C18 OBD column
[5 .mu.m, 19.times.50 mm; flow rate, 20 mL/min; Solvent A: H.sub.2O
with 0.1% HCO.sub.2H; Solvent B: CH.sub.3CN with 0.1% HCO.sub.2H;
injection volume: 4.0 mL (CH.sub.3CN); gradient: 10.fwdarw.100% B
over 10 min; mass-directed fraction collection]. Fractions with the
desired MW, were collected and concentrated on a RotaVap at rt to
remove most of the acetonitrile. The resulting mostly aqueous
solution was neutralized with pH 7 buffer and extracted with EtOAc.
The combined EtOAc extracts were dried (Na.sub.2SO.sub.4) and
concentrated to the desired product 4-3-1 (19.3 mg, 34% over 2
steps): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 15.63 (br s, 1H),
11.94 (br s, 1H), 8.67 (s, 1H), 7.51-7.48 (m, 2H), 7.42-7.32 (m,
3H), 7.28-7.22 (m, 3H), 7.18-7.16 (m, 2H), 5.38 (s, 2H), 5.22, 4.52
(ABq, J=11.6 Hz, 2H), 4.60 (d, J=6.7 Hz, 1H), 4.01 (d, J=9.8 Hz,
1H), 3.55 (s, 3H), 3.51-3.23 (m, 3H), 2.95-2.88 (m, 1H), 2.64-2.43
(m, 8H), 2.24-1.89 (m, 5H), 1.59-1.51 (m, 1H), 0.80 (s, 9H), 0.24
(s, 3H), 0.14 (s, 3H); MS (ESI) m/z 849.36 (M+H).
[0362] Aqueous HF (48-50%, 0.2 mL) was added to a solution of
Compound 4-3-1 (9 mg, 0.011 mmol, 1.0 equiv) in acetonitrile (0.5
mL) in a polypropylene reaction vessel at 23.degree. C. The
resulting mixture was stirred vigorously at 23.degree. C. overnight
and poured into aqueous K.sub.2HPO.sub.4 (2.5 g dissolved in 20 mL
water). The resulting mixture was extracted with EtOAc (30 mL, then
20 mL). The combined organic extracts were dried over anhydrous
sodium sulfate, filtered, and concentrated. The residue was used
directly in the next step without further purification.
[0363] The above crude product was dissolved in MeOH (2 mL) and
dioxane (0.5 mL). Pd--C (10 wt %, 5 mg) was added in one portion at
23.degree. C. The reaction vessel was sealed and purged with
hydrogen by briefly evacuating the flask followed by flushing with
hydrogen gas (1 atm). After stirring at 23.degree. C. for 25 min,
the resulting mixture was filtered through a small Celite pad. The
filtrate was concentrated. The residue was purified by a
preparative reverse phase HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100A column [10
.mu.m, 150.times.21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05 N
HCl/water; Solvent B: CH.sub.3CN; injection volume: 3.0 mL (0.05 N
HCl/water and MeCN, 1:1); gradient: 0.fwdarw.30% B over 10 min;
mass-directed fraction collection]. Fractions containing the
desired product, eluting at 7.5-8.7 min, were collected and
freeze-dried to yield Compound 142 (1.73 mg, some product was
spilled during the purification): .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.03 (s, 1H), 4.56 (t, J=6.8 Hz, 1H), 4.07 (s, 1H), 3.79
(s, 3H), 3.49-3.24 (m, 3H), 3.03-2.93 (m, 8H), 2.60-2.53 (m, 1H),
2.19-2.08 (m, 5H), 1.67-1.57 (m, 1H); MS (ESI) m/z 557.27
(M+H).
[0364] Compound 145.
##STR00261##
[0365] K.sub.2CO.sub.3 (43.9 mg, 0.318 mmol, 3.0 equiv) was added
in one portion to a solution of crude product 4-1-1 (0.106 mmol,
1.0 equiv) in a mixture of MeOH (2 mL), THF (0.5 mL) and water (0.3
mL). The resulting brownish reaction mixture was stirred at rt for
3 h, and diluted with saturated aqueous ammonium chloride and pH 7
phosphate buffer (1:1, 30 mL) and extracted with EtOAc (2.times.50
mL). The combined organic extracts were dried over anhydrous sodium
sulfate, filtered, and concentrated. The crude product 4-3-1 was
used directly in the next reaction. HCHO (47 .mu.L, 0.636 mmol, 6.0
equiv), acetic acid (18 .mu.L, 0.318 mmol, 3.0 equiv) and sodium
triacetoxyborohydride (45 mg, 0.212 mmol, 2.0 equiv) were added
sequentially to a solution of the above crude product 4-3-1 in
1,2-dichloroethane (3 mL) at 23.degree. C. After stirring for 35
min, the reaction mixture was quenched by the addition of saturated
aqueous sodium bicarbonate and pH 7 phosphate buffer (1:1, 30 mL)
and extracted with EtOAc (2.times.50 mL). The combined organic
extracts were dried over anhydrous sodium sulfate, filtered, and
concentrated. The residue was purified by a Preparative reverse
phase HPLC on a Waters Autopurification system using a Sunfire Prep
C18 OBD column [5 .mu.m, 19.times.50 mm; flow rate, 20 mL/min;
Solvent A: H.sub.2O with 0.1% HCO.sub.2H; Solvent B: CH.sub.3CN
with 0.1% HCO.sub.2H; injection volume: 4.0 mL (CH.sub.3CN);
gradient: 20.fwdarw.75% B over 20 min; mass-directed fraction
collection]. Fractions with the desired MW, eluting at 12.7-13.6
min, were collected and concentrated on a RotaVap at rt to remove
most of the acetonitrile. The resulting mostly aqueous solution was
neutralized with pH 7 phosphate buffer and extracted with EtOAc.
The combined EtOAc extracts were dried (Na.sub.2SO.sub.4) and
concentrated to the desired product 4-4-1 (44.5 mg, 48% over 3
steps): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 16.02 (br s, 1H),
10.09 (br s, 1H), 8.45 (s, 1H), 7.50-7.48 (m, 4H), 7.40-7.30 (m,
6H), 5.35 (s, 2H), 4.91, 4.80 (ABq, J=10.4 Hz, 2H), 3.99 (d, J=10.4
Hz, 1H), 3.87 (s, 3H), 3.32 (dd, J=4.9, 15.9 Hz, 1H), 3.05-2.91 (m,
3H), 2.56-2.41 (m, 8H), 2.36-2.23 (m, 5H), 2.14 (d, J=14.6 Hz, 1H),
1.95-1.89 (m, 1H), 1.78-1.68 (m, 1H), 1.62-1.53 (m, 1H), 1.39-1.33
(m, 1H), 0.81 (s, 9H), 0.26 (s, 3H), 0.12 (s, 3H); MS (ESI) m/z
863.56 (M+H).
[0366] Aqueous HF (48-50%, 0.3 mL) was added to a solution of
Compound 4-4-1 (44.5 mg, 0.051 mmol, 1.0 equiv) in acetonitrile
(0.8 mL) in a polypropylene reaction vessel at 23.degree. C. The
resulting mixture was stirred vigorously at 23.degree. C. overnight
and poured into aqueous K.sub.2HPO.sub.4 (3.6 g dissolved in 30 mL
water). The resulting mixture was extracted with EtOAc (2.times.25
mL). The combined organic extracts were dried over anhydrous sodium
sulfate, filtered, and concentrated. The residue was used directly
in the next step without further purification.
[0367] The above crude product was dissolved in MeOH (2 mL) and
EtOAc (1 mL). Pd--C (10 wt %, 22 mg) was added in one portion at
23.degree. C. The reaction vessel was sealed and purged with
hydrogen by briefly evacuating the flask followed by flushing with
hydrogen gas (1 atm). After stirring at 23.degree. C. for 1 h 20
min, more Pd--C (10 wt %, 10 mg) was added. The resulting mixture
was stirred under hydrogen for 2 h. Then s solution of HCl in MeOH
(0.5 N, 204 .mu.L, 2.0 equiv) was added. The resulting reaction
mixture was stirred under hydrogen for 45 min, and filtered through
a small Celite pad. The filtrate was concentrated. The residue was
purified by a preparative reverse phase HPLC on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu.
RP-.gamma. 100A column [10 .mu.m, 150.times.21.20 mm; flow rate, 20
mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH3CN; injection
volume: 3.0 mL (0.05 N HCl/water and MeCN, 1:1); gradient:
0.fwdarw.35% B over 10 min; mass-directed fraction collection].
Fractions containing the desired product, eluting at 7.3-8.2 min,
were collected and freeze-dried to yield Compound 145 (15.0 mg, 46%
yield over 2 steps): .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.02
(s, 1H), 4.36 (t, J=7.8 Hz, 1H), 4.07 (s, 1H), 3.79 (s, 3H),
3.78-3.75 (m, 1H), 3.28-3.24 (m, 2H), 3.03-2.93 (m, 11H), 2.73-2.67
(m, 1H), 2.30-2.10 (m, 5H), 1.67-1.57 (m, 1H); MS (ESI) m/z 571.36
(M+H).
[0368] Compound 143.
##STR00262##
[0369] DMF (0.5 mL) was added to a mixture of 2-6 (50 mg, 0.066
mmol, 1.0 equiv), pipecolinic acid (22 mg, 0.13 mmol, 2.0 equiv),
EDCI (25 mg, 0.13 mmol, 2.0 equiv) and HOBt (9 mg, 0.066 mmol, 1.0
equiv). The resulting red solution was stirred at rt overnight, and
purified by a Preparative reverse phase HPLC on a Waters
Autopurification system using a Sunfire Prep C18 OBD column [5
.mu.m, 19.times.50 mm; flow rate, 20 mL/min; Solvent A: H.sub.2O
with 0.1% HCO.sub.2H; Solvent B: CH.sub.3CN with 0.1% HCO.sub.2H;
injection volume: 4.0 mL (CH.sub.3CN); gradient: 10.fwdarw.100% B
over 10 min; mass-directed fraction collection]. Fractions with the
desired MW, eluting at 5.8-7.2 min, were collected and freeze-dried
to give the desired product 4-1-3 as a mixture of diastereomers
(25.9 mg, 45%): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 15.92 (s,
0.33H), 15.89 (s, 0.66H), 8.00 (s, 0.33H), 7.84 (s, 0.66 H),
7.50-7.48 (m, 2H), 7.41-7.29 (m, 8H), 5.36-5.34 (m, 2H), 4.90-4.73
(m, 2H), 4.00-3.97 (m, 1H), 3.85 (s, 1H), 3.72 (s, 2H), 3.52-3.23
(m, 3H), 2.95-2.82 (m, 2H), 2.61-2.43 (m, 8H), 2.34-2.12 (m, 3H),
1.88-1.38 (m, 5H), 0.83-0.80 (m, 9H), 0.26-0.24 (m, 3H), 0.15-0.13
(m, 3H); MS (ESI) m/z 863.47 (M+H).
[0370] Aqueous HF (48-50%, 0.2 mL) was added to a solution of the
above product 4-1-3 (13 mg, 0.015 mmol, 1.0 equiv) in acetonitrile
(0.5 mL) in a polypropylene reaction vessel at 23.degree. C. The
resulting mixture was stirred vigorously at 23.degree. C. overnight
and poured into aqueous K.sub.2HPO.sub.4 (2.5 g dissolved in 20 mL
water). The resulting mixture was extracted with EtOAc (40 mL, then
2.times.15 mL). The combined organic extracts were dried over
anhydrous sodium sulfate, filtered, and concentrated. The residue
was used directly in the next step without further
purification.
[0371] The above crude product was dissolved in MeOH (2 mL) and
dioxane (0.5 mL). Pd--C (10 wt %, 5 mg) was added in one portion at
23.degree. C. The reaction vessel was sealed and purged with
hydrogen by briefly evacuating the flask followed by flushing with
hydrogen gas (1 atm). After stirring at 23.degree. C. for 1 h 30
min, the reaction mixture was filtered through a small Celite pad.
The filtrate was concentrated. The residue was purified by a
preparative reverse phase HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100A column [10
.mu.m, 150.times.21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05 N
HCl/water; Solvent B: CH.sub.3CN; injection volume: 3.0 mL (0.05 N
HCl/water and MeCN, 1:1); gradient: 0.fwdarw.35% B over 10 min;
mass-directed fraction collection]. Fractions containing the
desired product, eluting at 8.5-9.3 min, were collected and
freeze-dried to yield Compound 143 (2.3 mg, 24% yield over 2
steps): .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.03 (s, 1H),
4.11-4.06 (m, 1H), 4.08 (s, 1H), 3.79 (s, 3H), 3.48-3.43 (m, 1H),
3.28-3.24 (m, 1H), 3.08-2.94 (m, 9H), 2.37-2.33 (m, 1H), 2.20-2.10
(m, 2H), 2.03-1.69 (m, 5H), 1.67-1.57 (m, 1H); MS (ESI) m/z 571.25
(M+H).
[0372] Compound 146.
##STR00263##
[0373] HCHO (6.7 .mu.L, 0.09 mmol, 6.0 equiv), acetic acid (5.2
.mu.L, 0.09 mmol, 6.0 equiv) and sodium triacetoxyborohydride (9.5
mg, 0.045 mmol, 3.0 equiv) were added sequentially to a solution of
Compound 4-1-3 in 1,2-dichloroethane (1 mL) at 23.degree. C. After
stirring for 2 h, the reaction mixture was quenched by the addition
of saturated aqueous sodium bicarbonate and brine (1:1, 10 mL) and
extracted with DCM (20 mL, then 10 mL). The combined organic
extracts were dried over anhydrous sodium sulfate, filtered, and
concentrated. The residue was purified by a Preparative reverse
phase HPLC on a Waters Autopurification system using a Sunfire Prep
C18 OBD column [5 .mu.m, 19.times.50 mm; flow rate, 20 mL/min;
Solvent A: H.sub.2O with 0.1% HCO.sub.2H; Solvent B: CH.sub.3CN
with 0.1% HCO.sub.2H; injection volume: 4.0 mL (CH.sub.3CN);
gradient: 10.fwdarw.100% B over 10 min; mass-directed fraction
collection]. Fractions with the desired MW, eluting at 7.1-7.6 min,
were collected and freeze-dried to afford the desired product 4-4-2
(6.8 mg, 51%): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 15.83 (s,
1H), 8.11-7.84 (m, 1H), 7.43-7.18 (m, 10H), 5.30 (s, 2H), 5.00-4.67
(m, 2H), 3.95-3.94 (m, 1H), 3.78-3.71 (m, 4H), 3.51-3.47 (m, 1H),
3.27-3.08 (m, 2H), 2.91-2.88 (m, 2H), 2.71-2.39 (m, 12H), 2.27-1.37
(m, 9H), 0.75 (s, 9H), 0.19 (m, 3H), 0.07-0.05 (m, 3H); MS (ESI)
m/z 877.44 (M+H).
[0374] Aqueous HF (48-50%, 0.2 mL) was added to a solution of the
above product 4-4-2 (6.8 mg, 0.0078 mmol, 1.0 equiv) in
acetonitrile (0.5 mL) in a polypropylene reaction vessel at
23.degree. C. The resulting mixture was stirred vigorously at
23.degree. C. overnight and poured into aqueous K.sub.2HPO.sub.4
(2.5 g dissolved in 20 mL water). The resulting mixture was
extracted with EtOAc (30 mL, then 20 mL). The combined organic
extracts were dried over anhydrous sodium sulfate, filtered, and
concentrated. The residue was used directly in the next step
without further purification.
[0375] The above crude product was dissolved in MeOH (2 mL) and
dioxane (0.5 mL). Pd--C (10 wt %, 2 mg) was added in one portion at
23.degree. C. The reaction vessel was sealed and purged with
hydrogen by briefly evacuating the flask followed by flushing with
hydrogen gas (1 atm). After stirring at 23.degree. C. for 2 h, the
reaction mixture was filtered through a small Celite pad. The
filtrate was concentrated. The residue was purified by a
preparative reverse phase HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100A column [10
.mu.m, 150.times.21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05 N
HCl/water; Solvent B: CH.sub.3CN; injection volume: 3.0 mL (0.05 N
HCl/water and MeCN, 1:1); gradient: 0.fwdarw.35% B over 10 min;
mass-directed fraction collection]. Fractions containing the
desired product, eluting at 8.2-8.6 min, were collected and
freeze-dried to yield Compound 146 (0.98 mg, 1:1 diastereomers):
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.03 (s, 0.5H), 8.01 (s,
0.5H), 4.08 (s, 1H), 4.08-4.03 (m, 1H), 3.80 (s, 3H), 3.58-3.55 (m,
1H), 3.26-3.24 (m, 1H), 3.20-3.12 (m, 1H), 3.03-2.92 (m, 11H),
2.34-2.31 (m, 1H), 2.20-2.10 (m, 2H), 2.03-1.1.97 (m, 2H),
1.92-1.81 (m, 2H), 1.67-1.61 (m, 2H); Fractions containing the
desired product, eluting at 8.6-9.1 min, were collected and
freeze-dried to yield Compound 4-5-2 (1.16 mg, single enantiomer):
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.01 (s, 1H), 4.08 (s,
1H), 4.08-4.04 (m, 1H), 3.80 (s, 3H), 3.58-3.55 (m, 1H), 3.28-3.24
(m, 1H), 3.20-3.12 (m, 1H), 3.03-2.92 (m, 11H), 2.35-2.32 (m, 1H),
2.21-2.10 (m, 2H), 2.00-1.1.97 (m, 2H), 1.92-1.82 (m, 2H),
1.67-1.61 (m, 2H); MS (ESI) m/z 585.27 (M+H).
[0376] Compounds 147 and 148 were prepared similarly to Compound
146 using enatiomerically pure pipecolinic acids.
[0377] Compound 147. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.01
(s, 1H), 4.10 (s, 1H), 4.12-4.08 (m, 1H), 3.80 (s, 3H), 3.58-3.55
(m, 1H), 3.29-3.24 (m, 1H), 3.22-3.15 (m, 1H), 3.04-2.93 (m, 11H),
2.34-2.31 (m, 1H), 2.23-2.18 (m, 1H), 2.12 (dd, J=13.7, 16.0 Hz,
1H), 1.99-1.97 (m, 2H), 1.92-1.78 (m, 2H), 1.72-1.56 (m, 2H); MS
(ESI) m/z 585.52 (M+H).
[0378] Compound 148. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.00
(s, 1H), 4.09 (s, 1H), 4.09-4.06 (m, 1H), 3.80 (s, 3H), 3.58-3.55
(m, 1H), 3.29-3.24 (m, 1H), 3.22-3.15 (m, 1H), 3.04-2.92 (m, 11H),
2.35-2.32 (m, 1H), 2.23-2.17 (m, 1H), 2.13 (dd, J=13.7, 16.0 Hz,
1H), 2.00-1.96 (m, 2H), 1.92-1.78 (m, 2H), 1.72-1.57 (m, 2H); MS
(ESI) m/z 585.56 (M+H).
[0379] Compound 144.
##STR00264##
[0380] A solution of 1-methylazepane-2-carbonyl chloride (HCl salt)
in DCM (0.4 M, 120 .mu.L, 0.048 mmol, 1.2 equiv) was added to a
solution of aniline 2-6 (30 mg, 0.040 mmol, 1.0 equiv) in THF (1
mL). The resulting light orange solution was stirred at rt for 10
min, and more acid chloride (0.4 M/DCM, 120 .mu.L, 0.048 mmol, 1.2
equiv) was added. The resulting mixture was stirred for 30 min and
diluted with brine and pH 7 phosphate buffer (1:1, 20 mL). The
resulting mixture was extracted with EtOAc (50 mL). The organic
phase was separated, dried (Na.sub.2SO.sub.4), filtered and
concentrated. The residue was purified by a Preparative reverse
phase HPLC on a Waters Autopurification system using a Sunfire Prep
C18 OBD column [5 .mu.m, 19.times.50 mm; flow rate, 20 mL/min;
Solvent A: H.sub.2O with 0.1% HCO.sub.2H; Solvent B: CH.sub.3CN
with 0.1% HCO.sub.2H; injection volume: 4.0 mL (CH.sub.3CN);
gradient: 20.fwdarw.100% B over 8 min; mass-directed fraction
collection]. Fractions with the desired MW, eluting at 6.3-9.0 min,
were collected and freeze-dried to afford the desired product 4-1-4
(20 mg, 84%): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 15.96 (s,
1H), 8.44 (s, 1H), 7.50-7.44 (m, 3H), 7.39-7.29 (m, 7H), 5.35 (s,
2H), 4.89, 4.80 (ABq, J=10.4 Hz, 2H), 3.98 4.80 (d, J=10.4 Hz, 1H),
3.86 (s, 3H), 3.32 (dd, J=4.9, 16.5 Hz, 1H), 3.09 (br s, 1H),
2.99-2.92 (m, 1H), 2.80-2.70 (m, 2H), 2.57-2.41 (m, 8H), 2.35-2.13
(m, 5H), 2.02-1.85 (m, 2H), 1.63-1.41 (m, 6H), 0.81 (s, 9H), 0.27
(s, 3H), 0.12 (s, 3H); MS (ESI) m/z 891.74 (M+H).
[0381] Aqueous HF (48-50%, 0.3 mL) was added to a solution of the
above product 4-1-4 (20 mg, 0.022 mmol, 1.0 equiv) in acetonitrile
(0.6 mL) in a polypropylene reaction vessel at 23.degree. C. The
resulting mixture was stirred vigorously at 23.degree. C. overnight
and poured into aqueous K.sub.2HPO.sub.4 (3.6 g dissolved in 25 mL
water). The resulting mixture was extracted with EtOAc (30 mL, then
20 mL). The combined organic extracts were dried over anhydrous
sodium sulfate, filtered, and concentrated. The residue was used
directly in the next step without further purification.
[0382] The above crude product was dissolved in MeOH (1 mL) and
HCl/MeOH (0.5 N, 88 .mu.L, 2.0 equiv). Pd--C (10 wt %, 8 mg) was
added in one portion at 23.degree. C. The reaction vessel was
sealed and purged with hydrogen by briefly evacuating the flask
followed by flushing with hydrogen gas (1 atm). After stirring at
23.degree. C. for 2 h 25 min, the reaction mixture was filtered
through a small Celite pad. The filtrate was concentrated. The
residue was purified by a preparative reverse phase HPLC on a
Waters Autopurification system using a Phenomenex Polymerx.TM.
10.mu. RP-.gamma. 100A column [10 .mu.m, 150.times.21.20 mm; flow
rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B:
CH.sub.3CN; injection volume: 3.0 mL (0.05 N HCl/water and MeCN,
1:1); gradient: 5.fwdarw.35% B over 10 min; mass-directed fraction
collection]. Fractions containing the desired product, eluting at
8.0-8.4 min, were collected and freeze-dried to yield Compound 144
(2.52 mg, 17% over 2 steps): .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.95 (s, 1H), 4.28 (dd, J=3.7, 6.4 Hz, 1H), 4.08 (s, 1H),
3.80 (s, 3H), 3.50 (t, J=5.0 Hz, 2H), 3.29-3.24 (m, 1H), 3.08-2.94
(m, 11H), 2.29-2.10 (m, 4H), 2.02-1.98 (m, 2H), 1.86-1.82 (m, 2H),
1.74-1.70 (m, 2H), 1.67-1.57 (m, 1H); MS (ESI) m/z 599.48
(M+H).
Example 5
[0383] Synthesis of Certain Compounds of Formula II, wherein Y is
--NH--C(O)-(saturated heterocyclyl). Scheme 5 depicts the synthesis
of still other compounds of Formula II, wherein Y is
--NH--C(O)-(saturated heterocyclyl).
##STR00265##
[0384] In Scheme 5, R.sup.A represents hydrogen,
(C.sub.1-C.sub.5)alkyl, --(C.sub.0-C.sub.5) alkylene-carbocyclyl,
or --(C.sub.0-C.sub.5)alkylene-heterocyclyl. For all of the
compounds made by Scheme 5 and described below, R.sup.Z is hydrogen
and R.sup.X and R.sup.Y are taken together with the carbon and
nitrogen atoms to which they are respectively bound to form an
optionally substituted 4-7 membered saturated heterocyclyl. It will
be readily apparent to those of skill in the art, however, that
this Scheme 5 will also be useful to synthesize compounds where
R.sup.X, R.sup.Y and R.sup.Z are R.sup.2, R.sup.5a and R.sup.5b,
respectively, as defined in structural formula (I).
[0385] Compound 149.
##STR00266##
[0386] A solution of (R)-N-(carbobenzoxy)prolyl chloride in DCM
(1.0 M, 69 .mu.L, 0.069 mmol, 1.2 equiv) was added to a solution of
aniline 2-6 (43.5 mg, 0.058 mmol, 1.0 equiv) in THF (1 mL). The
resulting light orange solution was stirred at rt for 3 h, diluted
with brine (20 mL). The resulting mixture was extracted with DCM
(2.times.25 mL). The combined organic extracts were dried
(Na.sub.2SO.sub.4), filtered and concentrated to yield the crude
product 5-1-1: MS (ESI) m/z 983.58 (M+H).
[0387] Aqueous HF (48-50%, 0.2 mL) was added to a solution of the
above crude product 5-1-1 in acetonitrile (0.5 mL) in a
polypropylene reaction vessel at 23.degree. C. The resulting
mixture was stirred vigorously at 23.degree. C. overnight and
poured into aqueous K.sub.2HPO.sub.4 (3.6 g dissolved in 30 mL
water). The resulting mixture was extracted with EtOAc (40 mL, then
2.times.15 mL). The combined organic extracts were dried over
anhydrous sodium sulfate, filtered, and concentrated. The residue
was used directly in the next step without further
purification.
[0388] The above crude product was dissolved in MeOH (2 mL) and
dioxane (0.4 mL). Pd--C (10 wt %, 20 mg) was added in one portion
at 23.degree. C. The reaction vessel was sealed and purged with
hydrogen by briefly evacuating the flask followed by flushing with
hydrogen gas (1 atm). After stirring at 23.degree. C. for 3 h, the
resulting reaction mixture was filtered through a small Celite pad.
The filtrate was concentrated. The residue was purified by a
preparative reverse phase HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100A column [10
.mu.m, 150.times.21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05 N
HCl/water; Solvent B: CH.sub.3CN; injection volume: 3.0 mL (0.05 N
HCl/water and MeCN, 1:1); gradient: 0.fwdarw.35% B over 10 min;
mass-directed fraction collection]. Fractions containing the
desired product, eluting at 7.5-8.4 min, were collected and
freeze-dried to yield Compound 149 (7.9 mg, 49% yield over 3
steps): .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.03 (s, 1H),
4.58 (dd, J=6.9, 8.7 Hz, 1H), 4.10 (s, 1H), 3.79 (s, 3H), 3.50-3.39
(m, 2H), 3.28-3.23 (m, 1H), 3.04-2.95 (m, 8H), 2.62-2.53 (m, 1H),
2.23-2.03 (m, 5H), 1.67-1.57 (m, 1H); MS (ESI) m/z 557.33
(M+H).
[0389] Compound 151. HCHO (14.4 .mu.L, 0.052 mmol, 6.0 equiv), TEA
(6.1 .mu.L, 0.044 mmol, 3.0 equiv) and sodium triacetoxyborohydride
(9.2 mg, 0.044 mmol, 3.0 equiv) were added sequentially to a
solution of Compound 149 (9.1 mg, 0.014 mmol, 1.0 equiv) in DMF
(0.3 mL) at 23.degree. C. After stirring for 15 min, the reaction
mixture was diluted with 0.05 N HCl/water, and purified by a
preparative reverse phase HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10 .mu. RP-.gamma. 100A column [10
.mu.m, 150.times.21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05 N
HCl/water; Solvent B: CH.sub.3CN; injection volume: 3.0 mL (0.05 N
HCl/water and MeCN, 1:1); gradient: 0.fwdarw.35% B over 10 min;
mass-directed fraction collection]. Fractions containing the
desired product, eluting at 8.0-8.8 min, were collected and
freeze-dried to yield Compound 5-3-1 mixed with a byproduct (MS
(ESI) m/z 601.35 (M+H)). The product was dissolved in MeOH (0.5
mL). Then conc. HCl (0.5 mL) was added. The resulting mixture was
stirred at rt for 40 min, and concentrated. The residue was diluted
with MeCN (1 mL) and freeze-dried. The residue was re-purified by a
preparative reverse phase HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100A column (same
method as before) to afford the desired product Compound 151(2.69
mg, 29% yield): .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.03 (s,
1H), 4.37 (t, J=7.8 Hz, 1H), 4.08 (s, 1H), 3.80 (s, 3H), 3.78-3.76
(m, 1H), 3.28-3.24 (m, 2H), 3.03-2.94 (m, 11H), 2.75-2.68 (m, 1H),
2.30-2.07 (m, 5H), 1.67-1.57 (m, 1H); MS (ESI) m/z 571.55
(M+H).
[0390] Compound 150. Compound 150 was prepared similarly to
Compound 149. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.12 (s,
1H), 5.28 (dd, J=7.3, 9.2 Hz, 1H), 4.21-4.14 (m, 1H), 4.09 (s, 1H),
4.07-4.00 (m, 1H), 3.81 (s, 3H), 3.28-3.24 (m, 1H), 3.04-2.90 (m,
9H), 2.71-2.62 (m, 1H), 2.23-2.09 (m, 2H), 1.66-1.57 (m, 1H); MS
(ESI) m/z 543.23 (M+H).
[0391] Compound 152. Compound 152 was prepared similarly to
Compound 151. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.10 (s,
1H), 5.18 (t, J=9.2 Hz, 1H), 4.21-4.04 (m, 3H), 3.80 (s, 3H),
3.26-3.25 (m, 1H), 3.05-2.87 (m, 12H), 2.69-2.59 (m, 1H), 2.22-2.10
(m, 2H), 1.68-1.57 (m, 1H); MS (ESI) m/z 557.35 (M+H).
Example 6
Synthesis of Certain Compounds of Formula II, wherein Y is
--NH--S(O).sub.m--(C.sub.1-C.sub.6 alkyl),
--NH--S(O).sub.m--(C.sub.1-C.sub.4 alkylene)-N(R.sup.2)(R.sup.3),
--NH--S(O).sub.m--N(R.sup.2)(R.sup.4),
--NH--S(O).sub.m-heterocyclyl, --NH--S(O).sub.m-carbocyclyl, and
--NH--S(O).sub.m--(C.sub.1-C.sub.4) alkylene-carbocyclyl
##STR00267##
[0393] In Scheme 6, R.sup.B represents --(C.sub.1-C.sub.6 alkyl),
--(C.sub.1-C.sub.4 alkylene)-N(R.sup.2)(R.sup.3),
--N(R.sup.2)(R.sup.4), -heterocyclyl, -carbocyclyl, or
--(C.sub.1-C.sub.4) alkylene-carbocyclyl.
[0394] Compound 153. Compound 2-6 (28 mg, 0.038 mmol) was dissolved
in CH.sub.2Cl.sub.2 (0.2 mL). Benzenesulfonyl chloride (14.4 .mu.L,
0.11 mmol, 3 eq) and pyridine (15.3 .mu.L, 0.19 mmol, 5 eq) were
added. The resulting mixture was stirred at rt and the reaction was
monitored by LC-MS. The reaction mixture was diluted with EtOAc
after SM was completely consumed. The organic solution was washed
with water, 1N HCl and brine, dried with Na.sub.2SO.sub.4 and
concentrated to give crude 6-1, where R is benzene (6-1-1).
Preparative reverse phase HPLC purification on a Waters
Autopurification system using a Sunfire Prep C18 OBD column [5
.mu.m, 19.times.50 mm; flow rate, 20 mL/min; Solvent A: H.sub.2O
with 0.1% HCO.sub.2H; Solvent B: CH.sub.3CN with 0.1% HCO.sub.2H;
injection volume: 4.0 mL (CH.sub.3CN); gradient: 80.fwdarw.100% B
over 15 min; mass-directed fraction collection]. Fractions with the
desired MW, eluting at 7.50-8.95 min, were collected and
concentrated on a RotaVap at rt to remove most of the acetonitrile.
The resulting mostly aqueous solution was extracted with EtOAc. The
combined EtOAc extracts were dried (Na.sub.2SO.sub.4) and
concentrated to give pure 6-1-1.
[0395] In a plastic vial, 6-1-1 was dissolved in CH.sub.3CN (1 mL).
Aqueous HF (48%, 0.25 mL) was added. After stirred at rt for 16 h,
the reaction mixture was poured into aqueous solution (12.5 mL) of
K.sub.2HPO.sub.4 (1.75 g). The resulting mixture was extracted
three times with EtOAc. The combined organic phases were washed
with brine, dried, concentrated to give crude product 6-2-1.
[0396] The crude 6-2-1 was dissolved in 0.5 N HCl in MeOH (155
.mu.L, 2 eq). The excess volatiles were evaporated. The pre-formed
HCl salt was re-dissolved in MeOH (2.0 mL) and to the resulting
solution was added palladium on carbon (10% wt, 9.0 mg, 30% w/w).
The reaction flask was briefly evacuated and re-filled with
hydrogen. The reaction mixture was stirred at rt and monitored by
LC-MS. After SM was consumed, the mixture was filtered through a
small pad of Celite. The filtrate was concentrated to give crude
Comopund 153, which was purified by HPLC on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu.
RP-.gamma. 100 R column [30.times.21.20 mm, 10 micron; flow rate,
20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH.sub.3CN;
injection volume: 4.0 mL (0.05 N HCl/water); gradient:
15.fwdarw.60% B over 15 min; mass-directed fraction collection].
Fractions with the desired MW, eluting at 13.50-14.65 min, were
collected and freeze-dried to give product as a yellow solid:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.78 (d, J=7.8 Hz, 2H),
7.56 (t, J=7.8 Hz, 1H), 7.45 (t, J=7.8 Hz, 2H), 7.38 (s, 1H), 4.02
(s, 1H), 3.76 (s, 3H), 3.30-3.15 (m, 1H), 3.10-2.90 (m, 8H),
2.18-2.01 (m, 2H), 1.62-1.52 (m, 1H); MS (ESI) m/z 600.2 (M+H),
calcd for C.sub.28H.sub.30N.sub.3O.sub.10S 600.16.
[0397] Compound 154. Compound 154 was obtained by the procedure of
Compound 153 employing methanesulfonyl chloride. Crude product was
purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 8.24-9.25 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.38 (s, 1H), 4.08 (s, 1H), 3.79 (s, 3H), 3.30-3.15 (m,
1H), 3.10-2.90 (m, 11H), 2.20-2.05 (m, 2H), 1.66-1.55 (m, 1H); MS
(ESI) m/z 538.5 (M+H), calcd for C.sub.23H.sub.28N.sub.3O.sub.10S
538.14.
[0398] Compound 155. Compound 155 was obtained by the procedure of
Compound 153 employing trifluoromethanesulfonic anhydride. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu., RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 20.fwdarw.70% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 7.70-8.58 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.28 (s, 1H), 4.08 (s, 1H), 3.79 (s, 3H), 3.30-3.15 (m,
1H), 3.10-2.90 (m, 8H), 2.20-2.10 (m, 2H), 1.66-1.55 (m, 1H); MS
(ESI) m/z 592.4 (M+H), calcd for
C.sub.23H.sub.25F.sub.3N.sub.3O.sub.10S 592.11.
[0399] Compound 156. Compound 156 was obtained by the procedure of
Compound 153 employing 3-methoxybenzenesulfonyl chloride. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 11.85-12.95 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.40-7.30 (m, 4H), 7.10 (br s, 1H), 4.08 (s, 1H), 3.76 (s,
6H), 3.25-3.10 (m, 1H), 3.00-2.85 (m, 8H), 2.18-2.00 (m, 2H),
1.66-1.55 (m, 1H); MS (ESI) m/z 630.4 (M+H), calcd for
C.sub.29H.sub.32N.sub.3O.sub.11S 630.17.
[0400] Compound 157. Compound 157 was obtained by the procedure of
Compound 153 employing 2-fluorobenzenesulfonyl chloride. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu., RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 11.84-12.74 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.77 (t, J=7.3 Hz, 1H), 7.65-7.56 (m 1H), 7.33 (s, 1H),
7.26-7.20 (m, 2H), 4.06 (s, 1H), 3.74 (s, 3H), 3.22-3.12 (m, 1H),
3.06-2.85 (m, 8H), 2.20-2.00 (m, 2H), 1.66-1.55 (m, 1H); MS (ESI)
m/z 618.4 (M+H), calcd for C.sub.28H.sub.29FN.sub.3O.sub.10S
618.15.
[0401] Compound 158. Compound 158 was obtained by the procedure of
Compound 153 employing p-nitrobenzenesulfonyl chloride. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 11.15-12.05 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.88 (d, J=7.8 Hz, 2H), 7.44-7.39 (m 3H), 4.09 (s, 1H),
3.79 (s, 3H), 3.22-3.12 (m, 1H), 3.06-2.85 (m, 8H), 2.20-2.00 (m,
2H), 1.66-1.55 (m, 1H); MS (ESI) m/z 615.4 (M+H), calcd for
C.sub.28H.sub.31N.sub.4O.sub.10S 615.17.
[0402] Compound 159. Compound 159 was obtained by the procedure of
Compound 153 employing 3-fluorobenzenesulfonyl chloride. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 14.00-15.10 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.60-7.43 (m, 3H), 7.37 (s, 1H), 7.32 (t, J=7.8 Hz, 1H),
4.07 (s, 1H), 3.78 (s, 3H), 3.22-3.12 (m, 1H), 3.08-2.88 (m, 8H),
2.20-2.00 (m, 2H), 1.62-1.52 (m, 1H); MS (ESI) m/z 618.4 (M+H),
calcd for C.sub.28H.sub.29FN.sub.3O.sub.10S 618.15.
[0403] Compound 160. Compound 160 was obtained by the procedure of
Compound 153 employing 4-methylbenzenesulfonyl chloride. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 13.80-15.30 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.63 (d, J=7.8 Hz, 2H), 7.35 (s, 1H), 7.23 (d, J=7.8 Hz,
2H), 4.06 (s, 1H), 3.74 (s, 3H), 3.25-3.10 (m, 1H), 3.06-2.85 (m,
8H), 2.34 (s, 3H), 2.18-2.00 (m, 2H), 1.60-1.50 (m, 1H); MS (ESI)
m/z 614.5 (M+H), calcd for C.sub.29H.sub.32N.sub.3O.sub.10S
614.17.
[0404] Compound 161. Compound 161 was obtained by the procedure of
Compound 153 employing 4-methoxybenzenesulfonyl chloride. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 13.20-15.00 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.69 (d, J=7.8 Hz, 2H), 7.36 (s, 1H), 6.93 (d, J=7.8 Hz,
2H), 4.07 (s, 1H), 3.80 (s, 3H), 3.76 (s, 3H), 3.25-3.12 (m, 1H),
3.08-2.85 (m, 8H), 2.18-2.00 (m, 2H), 1.61-1.52 (m, 1H); MS (ESI)
m/z 630.4 (M+H), calcd for C.sub.29H.sub.32N.sub.3O.sub.11S
630.17.
[0405] Compound 162. Compound 162 was obtained by the procedure of
Compound 153 employing 2-nitrobenzenesulfonyl chloride. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 13.00-14.50 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.62 (d, J=7.8 Hz, 1H), 7.55 (t, J=7.8 Hz, 1H), 7.31 (m,
2H), 7.14 (t, J=7.8 Hz, 1H), 4.08 (s, 1H), 3.78 (s, 3H), 3.22-3.12
(m, 1H), 3.06-2.85 (m, 8H), 2.22-2.00 (m, 2H), 1.61-1.51 (m, 1H);
MS (ESI) m/z 615.4 (M+H), calcd for
C.sub.28H.sub.31N.sub.4O.sub.10S 615.17.
[0406] Compound 163. Compound 163 was obtained by the procedure of
Compound 153 employing 3-nitrobenzenesulfonyl chloride. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 9.60-11.00 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.90 (d, J=7.8 Hz, 1H), 7.83 (s, 1H), 7.67-7.60 (m, 2H),
7.40 (s, 1H), 4.09 (s, 1H), 3.80 (s, 3H), 3.22-3.12 (m, 1H),
3.06-2.85 (m, 8H), 2.22-2.00 (m, 2H), 1.61-1.51 (m, 1H); MS (ESI)
m/z 615.4 (M+H), calcd for C.sub.28H.sub.31N.sub.4O.sub.10S
615.17.
[0407] Compound 164. Compound 164 was obtained by the procedure of
Compound 153 employing 1-methylimidazole-4-sulfonyl chloride. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.50% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 9.85-11.00 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 8.77 (s, 1H), 7.99 (s, 1H), 7.36 (s, 1H), 4.08 (s, 1H),
3.88 (s, 3H), 3.81 (s, 3H), 3.30-3.22 (m, 1H), 3.06-2.85 (m, 8H),
2.22-2.00 (m, 2H), 1.61-1.51 (m, 1H); MS (ESI) m/z 604.4 (M+H),
calcd for C.sub.26H.sub.30N.sub.5O.sub.10S 604.16.
[0408] Compound 165. Compound 165 was obtained by the procedure of
Compound 153 employing 1-methyl-1H-pyrazole-3-sulfonyl chloride.
Crude product was purified by HPLC on a Waters Autopurification
system using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R
column [30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent
A: 0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0
mL (0.05 N HCl/water); gradient: 15.fwdarw.50% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 11.85-13.00 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.64 (d, J=2.3 Hz, 1H), 7.45 (s, 1H), 6.62 (d, J=2.3 Hz,
1H), 4.07 (s, 1H), 3.88 (s, 3H), 3.77 (s, 3H), 3.22-3.14 (m, 1H),
3.06-2.85 (m, 8H), 2.22-2.00 (m, 2H), 1.63-1.53 (m, 1H); MS (ESI)
m/z 604.4 (M+H), calcd for C.sub.26H.sub.30N.sub.5O.sub.10S
604.16.
[0409] Compound 166. Compound 166 was obtained by the procedure of
Compound 153 employing isobutanesulfonyl chloride. Crude product
was purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 12.80-14.00 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.35 (s, 1H), 4.08 (s, 1H), 3.78 (s, 3H), 3.30-3.20 (m,
1H), 3.08-2.90 (m, 10H), 2.30-2.10 (m, 3H), 1.66-1.55 (m, 1H), 1.05
(d, J=6.8 Hz, 6H); MS (ESI) m/z 580.5 (M+H), calcd for
C.sub.26H.sub.34N.sub.3O.sub.10S 580.19.
[0410] Compound 167. Compound 167 was obtained by the procedure of
Compound 153 employing furan-2-sulfonyl chloride. Crude product was
purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 12.55-13.60 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.69 (s, 1H), 7.33 (s, 1H), 6.96 (d, J=3.6 Hz, 1H), 6.51
(t, J=1.8 Hz, 1H), 4.06 (s, 1H), 3.77 (s, 3H), 3.25-3.12 (m, 1H),
3.08-2.85 (m, 8H), 2.18-2.00 (m, 2H), 1.61-1.52 (m, 1H); MS (ESI)
m/z 590.4 (M+H), calcd for C.sub.26H.sub.28N.sub.3O.sub.11S
590.14.
[0411] Compound 168. Compound 168 was obtained by the procedure of
Compound 153 employing phenylmethanesulfonyl chloride. Crude
product was purified by HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 13.90-15.50 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.38-7.34 (m, 2H), 7.28-7.26 (m, 3H), 7.13 (s, 1H), 4.45
(s, 2H), 4.07 (s, 1H), 3.68 (s, 3H), 3.25-3.12 (m, 1H), 3.08-2.85
(m, 8H), 2.18-2.00 (m, 2H), 1.61-1.52 (m, 1H); MS (ESI) m/z 614.5
(M+H), calcd for C.sub.29H.sub.32N.sub.3O.sub.10S 614.17.
[0412] Compound 169. Compound 169 was obtained by the procedure of
Compound 153 employing ethanesulfonyl chloride. Crude product was
purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu., RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.50% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 12.05-13.20 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.36 (s, 1H), 4.08 (s, 1H), 3.78 (s, 3H), 3.30-3.20 (m,
1H), 3.10-2.85 (m, 10H), 2.22-2.05 (m, 2H), 1.65-1.55 (m, 1H), 1.36
(t, J=7.3 Hz, 3H); MS (ESI) m/z 552.4 (M+H), calcd for
C.sub.24H.sub.30N.sub.3O.sub.10S 552.16.
[0413] Compound 170. Compound 170 was obtained by the procedure of
Compound 153 employing 1-propanesulfonyl chloride. Crude product
was purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 11.75-12.80 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.35 (s, 1H), 4.08 (s, 1H), 3.78 (s, 3H), 3.30-3.20 (m,
1H), 3.10-2.85 (m, 10H), 2.22-2.05 (m, 2H), 1.88-1.82 (m, 2H),
1.65-1.55 (m, 1H), 1.00 (t, J=7.3 Hz, 3H); MS (ESI) m/z 566.5
(M+H), calcd for C.sub.25H.sub.32N.sub.3O.sub.10S 566.17.
[0414] Compound 171. Compound 171 was obtained by the procedure of
Compound 153 employing 1-butanesulfonyl chloride. Crude product was
purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 13.05-13.95 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.36 (s, 1H), 4.07 (s, 1H), 3.78 (s, 3H), 3.30-3.20 (m,
1H), 3.10-2.85 (m, 10H), 2.22-2.05 (m, 2H), 1.83-1.78 (m, 2H),
1.65-1.55 (m, 1H), 1.44-1.38 (m, 2H), 0.90 (t, J=7.3 Hz, 3H); MS
(ESI) m/z 580.5 (M+H), calcd for C.sub.26H.sub.34N.sub.3O.sub.10S
580.19.
[0415] Compound 172. Compound 172 was obtained by the procedure of
Compound 153 employing 1-hexanesulfonyl chloride. Crude product was
purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.65% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 14.85-15.30 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.37 (s, 1H), 4.08 (s, 1H), 3.79 (s, 3H), 3.30-3.20 (m,
1H), 3.10-2.85 (m, 10H), 2.22-2.05 (m, 2H), 1.85-1.78 (m, 2H),
1.65-1.55 (m, 1H), 1.44-1.25 (m, 6H), 0.88 (t, J=7.3 Hz, 3H); MS
(ESI) m/z 608.5 (M+H), calcd for C.sub.28H.sub.38N.sub.3O.sub.10S
608.22.
[0416] Compound 173. Compound 173 was obtained by the procedure of
Compound 153 employing 2,2,2-trifluoroethanesulfonyl chloride.
Crude product was purified by HPLC on a Waters Autopurification
system using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R
column [30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent
A: 0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0
mL (0.05 N HCl/water); gradient: 15.fwdarw.50% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 14.10-15.20 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.29 (s, 1H), 4.18 (q, J=9.6 Hz, 2H), 4.08 (s, 1H), 3.79
(s, 3H), 3.30-3.20 (m, 1H), 3.10-2.85 (m, 8H), 2.22-2.05 (m, 2H),
1.65-1.55 (m, 1H); MS (ESI) m/z 606.4 (M+H), calcd for
C.sub.24H.sub.27F.sub.3N.sub.3O.sub.10S 606.13.
[0417] Compound 174. Compound 174 was obtained by the procedure of
Compound 153 employing dimethylsulfamoyl chloride. Crude product
was purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 20.fwdarw.60% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 10.00-11.10 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.41 (s, 1H), 4.07 (s, 1H), 3.78 (s, 3H), 3.30-3.20 (m,
1H), 3.10-2.85 (m, 8H), 2.77 (s, 6H), 2.22-2.05 (m, 2H), 1.65-1.55
(m, 1H); MS (ESI) m/z 567.4 (M+H), calcd for
C.sub.24H.sub.31N.sub.4O.sub.10S 567.17.
[0418] Compound 175. Compound 175 was obtained by the procedure of
Compound 153 employing DMF as solvent. Crude product was purified
by HPLC on a Waters Autopurification system using a Phenomenex
Polymerx.TM. 10.mu. RP-.gamma. 100 R column [30.times.21.20 mm, 10
micron; flow rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent
B: CH.sub.3CN; injection volume: 4.0 mL (0.05 N HCl/water);
gradient: 5.fwdarw.30% B over 15 min; mass-directed fraction
collection]. Fractions with the desired MW, eluting at 11.90-13.05
min, were collected and freeze-dried to give product as a yellow
solid: .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.53 (s, 1H), 7.35
(s, 1H), 4.10 (s, 1H), 3.85 (s, 3H), 3.43 (s, 3H), 3.32 (s, 3H),
3.30-3.20 (m, 1H), 3.10-2.85 (m, 8H), 2.26-2.10 (m, 2H), 1.65-1.55
(m, 1H); MS (ESI) m/z 515.4 (M+H), calcd for
C.sub.25H.sub.31N.sub.4O.sub.8 515.21.
[0419] Compound 176. Compound 176 (9.0 mg, 0.0015 mmol) was
dissolved in MeOH (2.0 mL) and to the resulting solution was added
palladium on carbon (10% wt, 5.0 mg). The reaction flask was
briefly evacuated and re-filled with hydrogen. Formaldehyde (0.1
mL, 1.34 mmol) was then added. The reaction mixture was stirred at
rt and monitored by LC-MS. After SM was consumed, the mixture was
filtered through a small pad of Celite. The filtrate was
concentrated to give the crude product, which was purified by HPLC
on a Waters Autopurification system using a Phenomenex Polymerx.TM.
10.mu. RP-.gamma. 100 R column [30.times.21.20 mm, 10 micron; flow
rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B:
CH.sub.3CN; injection volume: 4.0 mL (0.05 N HCl/water); gradient:
15.fwdarw.60% B over 15 min; mass-directed fraction collection].
Fractions with the desired MW, eluting at 12.60-13.25 min, were
collected and freeze-dried to give product 6-3-24 as a yellow
solid: .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.66 (s, 1H),
7.58-7.45 (m, 3H), 7.40 (s, 1H), 4.06 (s, 1H), 3.79 (s, 3H),
3.25-3.15 (m, 1H), 3.12 (s, 6H), 3.06-2.90 (m, 8H), 2.26-2.10 (m,
2H), 1.65-1.55 (m, 1H) MS (ESI) m/z 643.5 (M+H), calcd for
C.sub.30H.sub.35N.sub.4O.sub.8S 643.20.
[0420] Compound 177. Compound 177 was obtained by the procedure of
Compound 153 employing pyridine-2-sulfonyl chloride: .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 8.60 (d, J=4.6 Hz, 1H), 7.95-8.10 (m,
2H), 7.65-7.75 (m, 1H), 7.48 (s, 1H), 4.04 (s, 1H), 3.79 (s, 3H),
2.75-3.50 (m, 9H), 2.00-2.20 (m, 2H), 1.52-1.70 (m, 1H); MS (ESI)
m/z 601.1 (M+H), calcd for C.sub.27H.sub.29N.sub.4O.sub.10S
601.16.
[0421] Compound 178. Compound 178 was obtained by the procedure of
Compound 153 employing pyridine-3-sulfonyl chloride: .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 7.35 (s, 1H), 4.08 (s, 1H), 3.80 (s,
3H), 2.80-3.90 (m, 14H), 1.90-2.40 (m, 5H), 1.55-1.80 (m, 2H); MS
(ESI) m/z 607.3 (M+H), calcd for C.sub.27H.sub.35N.sub.4O.sub.10S
607.21.
[0422] Compound 179.
##STR00268##
[0423] To a solution of 2-6 (38 mg, 0.05 mmol) in dichloroethane (1
mL) was added pyridine (6.1 .mu.L, 0.075 mmol, 1.5 equiv) and
2-chloroethanesulfonyl chloride (6.9 .mu.L, 0.65 mmol, 1.3 equiv).
After 18 h, the solution was heated to 45.degree. C. for 2 h, and
then was cooled to ambient temperature. Additional pyridine (12.2
.mu.L, 0.15 mmol, 3 equiv) and 2-chloroethanesulfonyl chloride
(15.7 .mu.L, 0.15 mmol, 3 equiv) were added and the reaction was
heated to 45.degree. C. for 21 h. Pyrrolidine (41 .mu.L, 0.500
mmol, 10 equiv) was added to the reaction mixture, and the reaction
was heated to 45.degree. C. After four hours, the reaction was
cooled, additional pyrrolidine was added (82 .mu.L, 1.00 mmol, 20
equiv), and the reaction was heated to 45.degree. C. for 24 h. The
reaction mixture was poured into aqueous pH 7 phosphate buffer (3
mL) and brine (2 mL), and the mixture was extracted with EtOAc
(2.times.10 mL). The combined organic extracts were dried
(Na.sub.2SO.sub.4), filtered, and concentrated to yield an oil.
[0424] Preparative reverse phase HPLC purification of this crude
mixture was performed on a Waters Autopurification system using a
Sunfire Prep C18 OBD column [5 .mu.m, 19.times.50 mm; flow rate, 20
mL/min; Solvent A: H.sub.2O with 0.1% HCO.sub.2H; Solvent B:
CH.sub.3CN with 0.1% HCO.sub.2H; injection volume: 4.times. 3.6-4.2
mL (CH3CN); gradient: 88.fwdarw.100% B over 12 min; mass-directed
fraction collection]. Fractions with the desired MW, eluting at
3.0-3.4 and 12.6-13.2 min, were collected and lyophilized to give
5.3 mg of 6-1-27a (12%) and 2.8 mg of 6-1-27b (6%),
respectively.
[0425] 6-1-27a: .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 15.9 (s,
1H), 8.18 (s, 1H), 7.52-7.30 (m, 11H), 5.41-5.31 (m, 2H), 4.95 (d,
J=10.4 Hz, 1H), 4.82 (d, J=10.4 Hz, 1H), 3.92 (d, J=10.4 Hz, 1H),
3.86 (s, 3H), 3.37-3.30 (m, 3H), 3.21-3.09 (m, 2H), 3.05-2.94 (m,
1H), 2.88-2.74 (m, 4H), 2.62-2.52 (m, 1H), 2.55-2.42 (m, 6H),
2.38-2.30 (m, 1H), 2.20-2.14 (m, 1H), 1.91-1.78 (m, 4H), 0.84 (s,
9H), 0.30 (s, 3H), 0.15 (s, 3H); MS (ESI) m/z 913.69 (M+H).
[0426] To a solution of 6-1-27a (5.3 mg, 0.0058 mmol, 1 equiv) in a
plastic vial in acetonitrile (1.0 mL) was added an aqueous solution
of hydrogen fluoride (50%, 250 .mu.L). After 20 h, the reaction
solution was diluted with an aqueous solution of K.sub.2HPO.sub.4
(3.2 g in 30 mL) and extracted with EtOAc (2.times.30 mL). The
combined organic layers were dried (Na.sub.2SO.sub.4), filtered,
and concentrated to yield the crude product. Palladium on carbon
(10%, 7 mg) was added to a solution of this crude oil in
dioxane:methanol (1:1, 1 mL). The flask was fitted with a septum
and evacuated and back-filled three times with hydrogen gas.
Hydrogen gas was bubbled through the reaction solution for three
minutes, and the reaction mixture was stirred under an atmosphere
(balloon) of hydrogen gas for 2 h. The reaction mixture was
filtered through Celite to remove the palladium catalyst and
concentrated under reduced pressure. Preparative reverse phase HPLC
purification of the resulting oil was performed on a Waters
Autopurification system using a Polymerx.TM. 10.mu. RP-.gamma. 100
R column [30.times.21.20 mm, 10 micron, solvent A: 0.05N HCl in
water, solvent B: CH.sub.3CN; injection volume: 4.8 mL (0.05N HCl
in water); gradient elution with 20.fwdarw.80% B over 15 min, then
held at 100% for 5 min; mass-directed fraction collection].
Fractions with the desired MW, eluting at 7.45-8.5 min, were
collected and freeze-dried to yield 0.65 mg of Compound 179 (18%):
.sup.1H NMR (400 MHz, CD.sub.3OD) 8 7.31 (s, 1H), 4.09 (s, 1H),
3.81-3.65 (m, 6H), 3.60-3.55 (m, 1H), 3.21-2.90 (m, 14H), 2.25-2.00
(m, 5H), 1.72-1.55 (m, 1H); MS (ESI) m/z 621.38 (M+H).
Example 7
Syntheis of Compounds of Formula II, wherein Y is --NH(R.sup.3),
and R.sup.3 is Selected from --(C.sub.1-C.sub.6)alkyl,
--(C.sub.1-C.sub.6) alkylene-carbocyclyl, and
--(C.sub.1-C.sub.6)alkylene-heterocyclyl
##STR00269##
[0428] In Scheme 7,--CH.sub.2--R.sup.C represents R.sup.3.
[0429] Compound 180. Compound 2-6 (20 mg, 0.026 mmol) was dissolved
in 1,2-dichloroethane (1.0 mL). Propionaldehyde (2.9 .mu.L, 0.040
mmol, 1.5 eq) was added, followed by acetic acid (7.6 .mu.L, 0.13
mmol, 5 eq). After stirred at rt for 1 h, sodium
triacetoxyborohydride (16.9 mg, 0.080 mmol, 3 eq) was added.
Stirring was continued for another 2 h. The reaction mixture was
poured into pH=7 buffer and sat. NaHCO.sub.3 solution, extracted
three times with EtOAc. The combined organic extracts were washed
with brine, dried over Na.sub.2SO.sub.4, and concentrated to give
crude 7-1, wherein --CH.sub.2-R.sup.A is n-propyl (7-1-1), which
was used directly for the next step without purification.
[0430] In a plastic vial, 7-1-1 was dissolved in CH.sub.3CN (1 mL).
Aqueous HF (48%, 0.25 mL) was added. After stirred at rt for 16 h,
the reaction mixture was poured into aqueous solution (12.5 mL) of
K.sub.2HPO.sub.4 (1.75 g). The resulting mixture was extracted
three times with CH.sub.2Cl.sub.2. The combined organic phases were
washed with brine, dried, concentrated to give crude 7-2, wherein
--CH.sub.2--R.sup.A is n-propyl (7-2-1).
[0431] The crude 7-2-1 was dissolved in 0.5 N HCl in MeOH (105
.mu.L, 2 eq). The excess volatiles were evaporated. The pre-formed
HCl salt was re-dissolved in MeOH (2.0 mL) and to the resulting
solution was added palladium on carbon (10% wt, 7.0 mg, 30% w/w).
The reaction flask was briefly evacuated and re-filled with
hydrogen. The reaction mixture was stirred at rt and monitored by
LC-MS. After SM was consumed, the mixture was filtered through a
small pad of Celite. The filtrate was concentrated to give the
crude product, which was purified by HPLC on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu.
RP-.gamma. 100 R column [30.times.21.20 mm, 10 micron; flow rate,
20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH.sub.3CN;
injection volume: 4.0 mL (0.05 N HCl/water); gradient:
15.fwdarw.60% B over 15 min; mass-directed fraction collection].
Fractions with the desired MW, eluting at 6.80-7.70 min, were
collected and freeze-dried to give Compound 180 as a yellow solid:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.30 (s, 1H), 4.10 (s,
1H), 3.86 (s, 3H), 3.40-3.25 (m, 3H), 3.10-2.90 (m, 8H), 2.26-2.16
(m, 2H), 1.85-1.75 (m, 2H), 1.62-1.52 (m, 1H), 1.06 (t, J=7.3 Hz,
3H); MS (ESI) m/z 502.4 (M+H), calcd for
C.sub.25H.sub.32N.sub.3O.sub.8 502.21.
[0432] Compound 181. Compound 181 was obtained by the procedure of
Compound 180 employing isovaleraldehyde. Crude product was purified
by HPLC on a Waters Autopurification system using a Phenomenex
Polymerx.TM. 10.mu. RP-.gamma. 100 R column [30.times.21.20 mm, 10
micron; flow rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent
B: CH.sub.3CN; injection volume: 4.0 mL (0.05 N HCl/water);
gradient: 15.fwdarw.50% B over 15 min; mass-directed fraction
collection]. Fractions with the desired MW, eluting at 10.40-11.75
min, were collected and freeze-dried to give product as a yellow
solid: .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.38 (s, 1H), 4.10
(s, 1H), 3.87 (s, 3H), 3.50-3.40 (m, 2H), 3.35-3.30 (m, 1H),
3.10-2.90 (m, 8H), 2.26-2.16 (m, 2H), 1.80-1.58 (m, 4H), 0.98 (d,
J=6.4 Hz, 6H); MS (ESI) m/z 530.4 (M+H), calcd for
C.sub.27H.sub.36N.sub.3O.sub.8 530.24.
[0433] Compound 182. Compound 182 was obtained by the procedure of
Compound 180 employing 2-methylbutyraldehyde. Crude product was
purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.50% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 11.80-12.90 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.37 (s, 1H), 4.10 (s, 1H), 3.86 (s, 3H), 3.40-3.20 (m,
3H), 3.10-2.90 (m, 8H), 2.26-2.16 (m, 2H), 1.95-1.85 (m, 1H),
1.70-1.55 (m, 2H), 1.40-1.30 (m, 1H), 1.12 (d, J=6.4 Hz, 3H), 0.98
(t, J=7.3 Hz, 3H); MS (ESI) m/z 530.4 (M+H), calcd for
C.sub.27H.sub.36N.sub.3O.sub.8 530.24.
[0434] Compound 183. Compound 183 was obtained by the procedure of
Compound 180 employing cyclopropanecarboxaldehyde. Crude product
was purified by HPLC on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100 R column
[30.times.21.20 mm, 10 micron; flow rate, 20 mL/min; Solvent A:
0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 4.0 mL
(0.05 N HCl/water); gradient: 15.fwdarw.50% B over 15 min;
mass-directed fraction collection]. Fractions with the desired MW,
eluting at 10.40-11.75 min, were collected and freeze-dried to give
product as a yellow solid: .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.37 (s, 1H), 4.10 (s, 1H), 3.87 (s, 3H), 3.35-3.30 (m,
3H), 3.10-2.90 (m, 8H), 2.26-2.16 (m, 2H), 1.70-1.58 (m, 1H),
1.25-1.16 (m, 1H), 0.76-0.70 (m, 2H), 0.46-0.42 (m, 2H); MS (ESI)
m/z 514.4 (M+H), calcd for C.sub.26H.sub.32N.sub.3O.sub.8
514.21.
[0435] Compound 184. Compound 2-6 (32 mg, 0.042 mmol) was dissolved
in 1,2-dichloroethane (1.0 mL). N-(tert-butoxycarbonyl)-L-prolinal
(12 .mu.L, 0.064 mmol, 1.5 eq) was added, followed by acetic acid
(12 .mu.L, 0.21 mmol, 5 eq). After stirred at rt for 1 h, sodium
triacetoxyborohydride (27 mg, 0.13 mmol, 3 eq) was added. Stirring
was continued for another 2 h. The reaction mixture was poured into
pH=7 buffer and sat. NaHCO3 solution, extracted three times with
EtOAc. The combined organic extracts were washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated to give crude 7-1, wherein
R.sup.A is pyrrolidin-2-yl (7-1-5), which was used directly for the
next step without purification.
[0436] Compound 7-1-5 obtained above was dissolved in anhydrous
dioxane (1 mL). Solution of HCl in dioxane (4M, 1 mL) was added at
rt. The resulting mixture was stirred at rt and the reaction was
monitored by LC-MS. The volatiles were evaporated after SM was
completely consumed. The residue was suspended in EtOAc, and washed
with sat. NaHCO.sub.3, brine, dried (Na.sub.2SO.sub.4), and
concentrated. Preparative reverse phase HPLC purification on a
Waters Autopurification system using a Sunfire Prep C18 OBD column
[5 .mu.m, 19.times.50 mm; flow rate, 20 mL/min; Solvent A: H.sub.2O
with 0.1% HCO.sub.2H; Solvent B: CH.sub.3CN with 0.1% HCO.sub.2H;
injection volume: 4.0 mL (CH.sub.3CN); gradient: 0.fwdarw.100% B
over 10 min; mass-directed fraction collection]. Fractions with the
desired MW, eluting at 7.08-8.00 min, were collected and
concentrated on a RotaVap at rt to remove most of the acetonitrile.
The resulting mostly aqueous solution was extracted with EtOAc. The
combined EtOAc extracts were dried (Na.sub.2SO.sub.4) and
concentrated.
[0437] The residue was dissolved in CH.sub.3CN (1 mL) in a plastic
vial. Aqueous HF (48%, 0.25 mL) was added. After stirred at rt for
16 h, the reaction mixture was poured into aqueous solution (12.5
mL) of K.sub.2HPO.sub.4 (1.75 g). The resulting mixture was
extracted three times with EtOAc. The combined organic phases were
washed with brine, dried, concentrated to give crude product 7-2,
wherein RA is pyrrolidin-2-yl (7-2-5).
[0438] The crude 7-2-5 was dissolved in 0.5 N HCl in MeOH (105
.mu.L, 3 eq). The excess volatiles were evaporated. The pre-formed
HCl salt was re-dissolved in MeOH (2.0 mL) and to the resulting
solution was added palladium on carbon (10% wt, 7.0 mg, 30% w/w).
The reaction flask was briefly evacuated and re-filled with
hydrogen. The reaction mixture was stirred at rt and monitored by
LC-MS. After SM was consumed, the mixture was filtered through a
small pad of Celite. The filtrate was concentrated to give crude
Compound 184, which was purified by HPLC on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu.
RP-.gamma. 100 R column [30.times.21.20 mm, 10 micron; flow rate,
20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH.sub.3CN;
injection volume: 4.0 mL (0.05 N HCl/water); gradient: 10-40% B
over 15 min; mass-directed fraction collection]. Fractions with the
desired MW, eluting at 9.95-10.05 min, were collected and
freeze-dried to give product as a yellow solid: .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 7.30 (s, 1H), 4.10 (s, 1H), 3.86 (s, 3H),
3.40-3.25 (m, 3H), 3.10-2.90 (m, 8H), 2.26-2.16 (m, 2H), 1.85-1.75
(m, 2H), 1.62-1.52 (m, 1H), 1.06 (t, J=7.3 Hz, 3H); MS (ESI) m/z
543.4 (M+H), calcd for C.sub.27H.sub.35N.sub.4O.sub.8 543.24.
[0439] Compound 185. Compound 185 was obtained by the procedure of
Compound 180 employing 3,3-dimethylbutyraldehyde. .sup.1H NMR (400
MHz, CD.sub.3OD) 87.24 (s, 1H), 4.09 (s, 1H), 3.85 (s, 3H),
3.35-3.45 (m, 2H), 2.95-3.10 (m, 9H), 2.12-2.25 (m, 2H), 1.65-1.72
(m, 3H), 0.98 (s, 9H); MS (ESI) m/z 544.4 (M+H), calcd for
C.sub.28H.sub.38N.sub.3O.sub.8 544.23.
[0440] Compound 186. Compound 186 was obtained by the procedure of
Compound 180 employing tert-butyl-N(Cbz)CH.sub.2CHO. .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 6.95 (s, 1H), 4.07 (s, 1H), 3.81 (s,
3H), 3.64 (t, J=6.5 Hz, 2H), 3.20 (dd, J=3.9, 14.2 Hz, 1H), 3.03
(s, 3H), 2.95 (s, 3H), 2.90-3.10 (m, 4H), 2.15-2.20 (m, 1H),
2.01-2.13 (m, 1H), 1.55-1.65 (m, 1H), 1.41 (s, 9H); MS (ESI) m/z
559.2 (M+H), calcd for C.sub.28H.sub.39N.sub.4O.sub.8 559.28.
Example 8
Synthesis of Compounds of Formula II, wherein Y is
--(C.sub.1-C.sub.4)alkylene-N(R.sup.2)(R.sup.3),
--CH.dbd.N--OR.sup.2, or --C(O)--N(R.sup.2)(R.sup.4)
##STR00270##
[0442] Phenyl 2-(benzyloxy)-3-bromo-5-methoxy-6-methylbenzoate
(8-2). A solution of Br.sub.2 (0.328 mL, 6.4 mmol, 1.1 equiv) in
HOAc (2 mL) was added to a solution of phenol 1-4 (1.50 g, 5.8
mmol, 1.0 equiv) in HOAc (10 mL) dropwise at 10.degree. C. The
resulting red solution was then stirred at rt for 50 min, and more
Br.sub.2 (30 .mu.L, 0.58 mmol, 0.1 equiv) in HOAc (0.2 mL) was
added. After stirring at rt for 1 h, the reaction mixture was
poured onto ice-water (80 mL), diluted with EtOAc (150 mL). The
organic phase was separated, washed with water (4.times.75 mL). The
resulting organic phase was dried over magnesium sulfate, filtered,
and concentrated to afford a brownish solid. The crude product was
used directly for the next reaction. K.sub.2CO.sub.3 (1.60 g, 11.6
mmol, 2.0 equiv) was added to a solution of the above product in
acetone (30 mL). Then BnBr (1.03 mL, 8.7 mmol, 1.5 equiv) was
added. The resulting reaction mixture was stirred at rt for 20 h
and heated at 50.degree. C. for 45 min. The reaction was then
cooled to rt and filtered. The filtrate was concentrated, and the
residue was diluted with EtOAc and water. The organic phase was
separated, washed with brine. The resulting organic phase was dried
over magnesium sulfate, filtered, and concentrated. The residue was
purified by flash chromatography (2-4% EtOAc/Hexanes) to give 8-2
(1.79 g, 72% over 2 steps): .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.51-7.49 (m, 2H), 7.39-7.33 (m, 5H), 7.26-7.22 (m, 1H),
7.10 (s, 1H), 7.07-7.05 (m, 2H), 5.08 (s, 2H), 3.85 (s, 3H), 2.26
(s, 3H); MS (ESI) m/z 425.26, 427.32 (M-H).
[0443] Phenyl 2-(benzyloxy)-3-formyl-5-methoxy-6-methylbenzoate
(8-3). To a solution of 8-2 (76 mg, 0.178 mmol, 1.0 equiv) in
anhydrous THF (1 mL) was added a solution of i-PrMgCl.LiCl in THF
(1.2 M, 252 .mu.L, 0.303 mmol, 1.7 equiv) dropwise at 0.degree. C.
under a N.sub.2 atmosphere. The resulting reaction mixture was
stirred at rt for 1 h 20 min. DMF (47 .mu.L, 0.606 mmol, 3.0 equiv)
was then added. The reaction was stirred for 5 min at rt, and
quenched by saturated aqueous NH.sub.4Cl. The resulting mixture was
extracted twice with EtOAc (30 mL). The organic phase was
separated, dried over sodium sulfate, filtered, and concentrated.
The residue was purified by flash chromatography (2-5%
EtOAc/Hexanes) to give 8-3 (55.5 mg, 83%): .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 10.14 (s, 1H), 7.41-7.34 (m, 7H), 7.33 (s, 1H),
7.28-7.25 (m, 1H), 7.13-7.11 (m, 2H), 5.12 (s, 2H), 3.89 (s, 3H),
2.39 (s, 3H); MS (ESI) m/z 375.33 (M-H).
[0444] Phenyl
2-(benzyloxy)-3-((tert-butylimino)methyl)-5-methoxy-6-methylbenzoate
(8-4). Tert-butyl amine (78 .mu.L, 0.738 mmol, 5.0 equiv) was added
to a solution of 8-3 (55.5 mg, 0.147 mmol, 1.0 equiv) in toluene (1
mL). The resulting reaction mixture was stirred at rt overnight,
and then diluted with toluene (5 mL), concentrated, and dried under
high vacuum. The crude product 8-4 was used directly for the next
reaction. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.63 (s, 1H),
7.64 (s, 1H), 7.50-7.44 (m, 7H), 7.38-7.34 (m, 1H), 7.24-7.22 (m,
2H), 5.09 (s, 2H), 4.02 (s, 3H), 2.45 (s, 3H), 1.34 (s, 9H); MS
(ESI) m/z 432.39 (M+H).
[0445]
(4aS,11aR,12aS,13S)-3,7-bis(benzyloxy)-4a-(tert-butyldimethylsilylo-
xy)-13-(dimethylamino)-5-hydroxy-10-methoxy-4,6-dioxo-4,4a,6,11,11a,12,12a-
,13-octahydrotetraceno[2,3-d]isoxazole-8-carbaldehyde (8-5). A
solution of the above crude imine 8-4 in THF (1.5 mL) was added to
a solution of LDA (1.8 M, 90 .mu.L, 0.162 mmol, 1.1 equiv) and
TMEDA (24 .mu.L, 0.162 mmol, 1.1 equiv) in THF (1 mL) dropwise via
a cannula at -78.degree. C. The resulting red solution was stirred
at that temperature for 5 min. A solution of enone 1-6 (57 mg,
0.118 mmol, 0.8 equiv) in THF (2 mL) was added very slowly. The
resulting light yellow solution was stirred at -78.degree. C. for 5
min. A solution of LHMDS in THF (1.0 M, 147 .mu.L, 0.147 mmol, 1.0
equiv) was added. The resulting reaction mixture was then allowed
to warm up to -20.degree. C. over 40 min. Saturated aqueous
NH.sub.4Cl was added. The resulting mixture was stirred at rt for 5
min, and extracted with EtOAc (30 mL). The organic phase was
separated, dried over sodium sulfate, filtered, and concentrated.
The residue was purified by a Preparative reverse phase HPLC on a
Waters Autopurification system using a Sunfire Prep C18 OBD column
[5 .mu.m, 19.times.50 mm; flow rate, 20 mL/min; Solvent A: H.sub.2O
with 0.1% HCO.sub.2H; Solvent B: CH.sub.3CN with 0.1% HCO.sub.2H;
injection volume: 4.0 mL (CH.sub.3CN); gradient: 10.fwdarw.100% B
over 10 min; mass-directed fraction collection]. Fractions with the
desired MW of the imine product and aldehyde product were
collected, allowed to stand in the hood overnight, and concentrated
to afford the desired aldehyde product 8-5 (62.4 mg, 69%): .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 15.86 (br s, 1H), 10.12 (s, 1H),
7.50-7.48 (m, 2H), 7.39-7.29 (m, 9H), 5.36 (s, 2H), 5.02, 4.93
(ABq, J=11.0 Hz, 2H), 3.97 (d, J=11.0 Hz, 1H), 3.88 (s, 3H), 3.42
(dd, J=4.9, 16.5 Hz, 1H), 3.04-2.96 (m, 1H), 2.59-2.56 (m, 1H),
2.52-2.44 (m, 7H), 2.37 (t, J=15.9 Hz, 1H), 2.18 (d, J=14.6 Hz,
1H), 0.82 (s, 9H), 0.28 (s, 3H), 0.14 (s, 3H); MS (ESI) m/z 765.52
(M+H).
[0446] Compound 190. A solution of methylamine in THF (2.0 M, 122
.mu.L, 0.24 mmol, 6.0 equiv), acetic acid (14 .mu.L, 0.24 mmol, 6.0
equiv) and sodium triacetoxyborohydride (17 mg, 0.08 mmol, 2.0
equiv) were added sequentially to a solution of compound 8-5 (31
mg, 0.040 mmol, 1.0 equiv) in 1,2-dichloroethane (1 mL) at
23.degree. C. After stirring for 3 h, the reaction mixture was
quenched by the addition of saturated aqueous sodium bicarbonate (2
mL) and pH 7 phosphate buffer (10 mL) and extracted with
dichloromethane (2.times.25 mL). The combined organic extracts were
dried over anhydrous sodium sulfate, filtered, and concentrated.
The residue was used directly in the next reaction.
[0447] Aqueous HF (48-50%, 0.3 mL) was added to a solution of the
above crude product in acetonitrile (0.6 mL) in a polypropylene
reaction vessel at 23.degree. C. The resulting mixture was stirred
vigorously at 23.degree. C. overnight and poured into aqueous
K.sub.2HPO.sub.4 (3.6 g dissolved in 25 mL water). The resulting
mixture was extracted with EtOAc (30 mL, then 20 mL). The combined
organic extracts were dried over anhydrous sodium sulfate,
filtered, and concentrated. The residue was used directly in the
next step without further purification.
[0448] The above crude product was dissolved in MeOH (1 mL) and
HCl/MeOH (0.5 N, 80 .mu.L, 2.0 equiv). Pd--C (10 wt %, 10 mg) was
added in one portion at 23.degree. C. The reaction vessel was
sealed and purged with hydrogen by briefly evacuating the flask
followed by flushing with hydrogen gas (1 atm). After stirring at
23.degree. C. for 40 min, the reaction mixture was filtered through
a small Celite pad. The filtrate was concentrated. The residue was
purified by a preparative reverse phase HPLC on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu.
RP-.gamma. 100A column [10 .mu.m, 150.times.21.20 mm; flow rate, 20
mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH.sub.3CN;
injection volume: 3.0 mL (0.05 N HCl/water and MeCN, 1:1);
gradient: 15.fwdarw.35% B over 15 min; mass-directed fraction
collection]. Fractions containing the desired product, eluting at
5.2-7.1 min, were collected and freeze-dried to yield Compound 190
(11.6 mg, 52% over 3 steps): .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.39 (s, 1H), 4.25 (s, 2H), 4.11 (s, 1H), 3.84 (s, 3H),
3.32-3.27 (m, 1H), 3.04-2.96 (m, 8H), 2.73 (s, 3H), 2.24-2.20 (m,
1H), 2.14 (dd, J=13.7, 16.0 Hz, 1H), 1.66-1.57 (m, 1H); MS (ESI)
m/z 488.35 (M+H).
[0449] The following compounds were prepared similarly to Compound
190 coupling the appropriate amine NHR.sup.2R.sup.3 to 8-5.
[0450] Compound 191. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.43
(s, 1H), 4.28 (s, 2H), 4.11 (s, 1H), 3.85 (s, 3H), 3.33-3.28 (m,
1H), 3.05-2.97 (m, 8H), 2.92 (d, J=7.3 Hz, 2H), 2.25-2.06 (m, 3H),
1.67-1.58 (m, 1H), 1.05 (d, J=6.4 Hz, 6H); MS (ESI) m/z 530.31
(M+H).
[0451] Compound 192. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.42
(s, 1H), 4.33 (s, 2H), 4.10 (s, 1H), 3.85 (s, 3H), 3.34-3.29 (m,
1H), 3.04-2.96 (m, 8H), 2.87 (s, 2H), 2.24-2.13 (m, 2H), 1.68-1.58
(m, 1H), 1.05 (s, 9H); MS (ESI) m/z 544.55 (M+H).
[0452] Compound 193. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.46
(s, 1H), 4.64-4.60 (m, 1H), 4.18 (d, J=12.8 Hz, 1H), 4.12 (s, 1H),
3.86 (s, 3H), 3.33-3.30 (m, 1H), 3.16-2.97 (m, 10H), 2.84 (s, 3H),
2.30-2.12 (m, 3H), 1.67-1.58 (m, 1H), 1.12-1.05 (m, 6H); MS (ESI)
m/z 544.34 (M+H).
[0453] Compound 194. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.54
(s, 1H), 4.63-4.58 (m, 1H), 4.30 (d, J=12.8 Hz, 1H), 4.11 (s, 1H),
3.86 (s, 3H), 3.34-3.27 (m, 1H), 3.11 (dd, J=1.8, 13.7 Hz, 1H),
3.04-2.96 (m, 12H), 2.24-2.11 (m, 2H), 1.67-1.58 (m, 1H), 1.07 (d,
J=5.5 Hz, 9H); MS (ESI) m/z 558.35 (M+H).
[0454] Compound 195. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.43
(s, 1H), 4.37 (s, 2H), 4.11 (s, 1H), 3.85 (s, 3H), 3.33-3.28 (m,
1H), 3.04-2.96 (m, 8H), 2.90 (s, 3H), 2.89 (s, 3H), 2.24-2.21 (m,
1H), 2.14 (dd, J=14.2, 16.5 Hz, 1H), 1.66-1.56 (m, 1H); MS (ESI)
m/z 502.37 (M+H).
[0455] Compound 196. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.42
(s, 1H), 4.18-4.27 (m, 2H), 4.09 (s, 1H), 3.86 (s, 3H), 3.78-3.90
(m, 2H), 2.95-3.40 (m, 7H), 2.10-2.25 (m, 2H), 1.55-1.70 (m, 1H),
1.48 (s, 9H); MS (ESI) m/z 530.3 (M+H).
[0456] Compound 197.
##STR00271##
[0457] Aqueous HF (48-50%, 0.2 mL) was added to a solution of
aldehyde 8-5 (24 mg, 0.031 mmol, 1.0 equiv) in acetonitrile (0.5
mL) in a polypropylene reaction vessel at 23.degree. C. The
resulting mixture was stirred vigorously at 23.degree. C. overnight
and poured into aqueous K.sub.2HPO.sub.4 (3.6 g dissolved in 25 mL
water). The resulting mixture was extracted with EtOAc (30 mL, then
20 mL). The combined organic extracts were dried over anhydrous
sodium sulfate, filtered, and concentrated. The residue was used
directly in the next step without further purification.
[0458] The above crude product was dissolved in MeOH (1 mL) and
HCl/MeOH (0.5 N, 61 .mu.L, 1.0 equiv). Pd--C (10 wt %, 5 mg) was
added in one portion at 23.degree. C. The reaction vessel was
sealed and purged with hydrogen by briefly evacuating the flask
followed by flushing with hydrogen gas (1 atm). After stirring at
23.degree. C. for 30 min, the reaction mixture was filtered through
a small Celite pad. The filtrate was concentrated. The residue was
purified by a preparative reverse phase HPLC on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu.
RP-.gamma. 100A column [10 .mu.m, 150.times.21.20 mm; flow rate, 20
mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH3CN; injection
volume: 3.0 mL (0.05 N HCl/water and MeCN, 1:1); gradient:
20.fwdarw.50% B over 15 min; mass-directed fraction collection].
Fractions containing the desired product, eluting at 11.0-12.1 min,
were collected and freeze-dried to yield compound 8-12 (4.3 mg, 29%
over 2 steps): .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.38 (s,
1H), 5.66 (s, 1H), 4.07 (s, 1H), 3.79 (s, 3H), 3.32-3.29 (m, 1H),
3.03-2.93 (m, 8H), 2.20-2.10 (m, 2H), 1.66-1.57 (m, 1H); MS (ESI)
m/z 473.29 (M+H).
[0459] Compound 197. NH.sub.2OH.HCl (1.3 mg, 0.018 mmol, 1.0 equiv)
was added to a solution of aldehyde 8-12 (4.3 mg, 0.009 mmol, 1.0
equiv) in MeOH (0.5 mL). The resulting reaction mixture was stirred
at rt for 1 h 30 min, and concentrated. The residue was purified by
a preparative reverse phase HPLC on a Waters Autopurification
system using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100A
column [10 .mu.m, 150.times.21.20 mm; flow rate, 20 mL/min; Solvent
A: 0.05 N HCl/water; Solvent B: CH.sub.3CN; injection volume: 3.0
mL (0.05 N HCl/water and MeCN, 1:1); gradient: 20.fwdarw.50% B over
15 min; mass-directed fraction collection]. Fractions containing
the desired product, eluting at 9.5-10.8 min, were collected and
freeze-dried to yield Compound 197 (2.0 mg, 45%): .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
8.41 (s, 1H), 7.60 (s, 1H), 4.07 (s, 1H), 3.81 (s, 3H), 3.32-3.30
(m, 1H), 3.03-2.93 (m, 8H), 2.20-2.11 (m, 2H), 1.67-1.57 (m, 1H);
MS (ESI) m/z 488.29 (M+H).
[0460] Compound 198. Compound 198 was prepared similarly to
compound 197: .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.40 (s,
1H), 7.59 (s, 1H), 4.08 (s, 1H), 3.94 (s, 3H), 3.81 (s, 3H),
3.32-3.26 (m, 1H), 3.04-2.93 (m, 8H), 2.23-2.10 (m, 2H), 1.67-1.57
(m, 1H); MS (ESI) m/z 502.34 (M+H).
Compound 189
##STR00272##
[0462] A solution of sulfamic acid (11 mg, 0.113 mmol, 1.4 equiv)
in water (0.35 mL) was added dropwise to a solution of aldehyde 8-5
(61.8 mg, 0.081 mmol, 1.0 equiv) in MeCN (7 mL) at 0.degree. C.
Then a solution of NaOClO in water (0.35 mL, 0.113 mmol, 1.4 equiv)
was added dropwise. The resulting reaction mixture was stirred at
0.degree. C. for 15 min, and diluted with water (5 mL). The
resulting mixture was stirred at rt for 5 min and extracted with
EtOAc (50 mL). The organic phase was separated, dried over sodium
sulfate, filtered and concentrated. The residue was purified by a
Preparative reverse phase HPLC on a Waters Autopurification system
using a Sunfire Prep C18 OBD column [5 .mu.m, 19.times.50 mm; flow
rate, 20 mL/min; Solvent A: H.sub.2O with 0.1% HCO.sub.2H; Solvent
B: CH.sub.3CN with 0.1% HCO.sub.2H; injection volume: 4.0 mL
(CH.sub.3CN); gradient: 50.fwdarw.100% B over 10 min; mass-directed
fraction collection]. Fractions with the desired MW, eluting at
9.0-10.6 min, were collected and concentrated to give the desired
product 8-6 (36.9 mg, 58%): .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 15.83 (s, 1H), 7.72 (s, 1H), 7.51-7.49 (m, 2H), 7.40-7.33
(m, 8H), 5.36 (s, 2H), 5.05, 4.95(ABq, J=9.8 Hz, 2H), 3.96 (d,
J=11.0 Hz, 1H), 3.90 (s, 3H), 3.42 (dd, J=4.9, 17.1 Hz, 1H),
3.04-2.97 (m, 1H), 2.61-2.58 (m, 1H), 2.53-2.45 (m, 7H), 2.37 (t,
J=16.5 Hz, 1H), 2.19 (d, J=14.6 Hz, 1H), 0.82 (s, 9H), 0.28 (s,
3H), 0.14 (s, 3H); MS (ESI) m/z 781.51 (M+H).
[0463] Compound 189. Aqueous HF (48-50%, 0.2 mL) was added to a
solution of aldehyde 8-6 (18 mg, 0.023 mmol, 1.0 equiv) in
acetonitrile (0.5 mL) in a polypropylene reaction vessel at
23.degree. C. The resulting mixture was stirred vigorously at
23.degree. C. overnight and poured into aqueous K.sub.2HPO.sub.4
(2.4 g dissolved in 25 mL water). The resulting mixture was
extracted with EtOAc (40 mL). The organic phase was dried over
anhydrous sodium sulfate, filtered, and concentrated. The residue
was used directly in the next step without further
purification.
[0464] The above crude product was dissolved in MeOH (2 mL) and
dioxane (2 mL). Pd--C (10 wt %, 8 mg) was added in one portion at
23.degree. C. The reaction vessel was sealed and purged with
hydrogen by briefly evacuating the flask followed by flushing with
hydrogen gas (1 atm). After stirring at 23.degree. C. for 40 min,
the reaction mixture was filtered through a small Celite pad. The
filtrate was concentrated. The residue was purified by a
preparative reverse phase HPLC on a Waters Autopurification system
using a Phenomenex Polymerx.TM. 10.mu. RP-.gamma. 100A column [10
.mu.m, 150.times.21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05 N
HCl/water; Solvent B: CH.sub.3CN; injection volume: 3.0 mL (0.05 N
HCl/water); gradient: 20.fwdarw.60% B over 15 min; mass-directed
fraction collection]. Fractions containing the desired product,
eluting at 6.6-8.0 min, were collected and freeze-dried to yield
Compound 189 (6.5 mg, 58% over 2 steps): .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.67 (s, 1H), 4.07 (s, 1H), 3.83 (s, 3H),
3.33-3.30 (m, 1H), 3.03-2.92 (m, 8H), 2.20-2.10 (m, 2H), 1.65-1.55
(m, 1H); MS (ESI) m/z 489.28 (M+H).
[0465] Compound 187.
##STR00273##
[0466] Neopentyl amine (9.2 .mu.L, 0.039 mmol, 2.0 equiv) was added
to a solution of 8-6 (15.3 mg, 0.020 mmol, 1.0 equiv), EDCI (7.5
mg, 0.039 mmol, 2.0 equiv) and HOBt (1.3 mg, 0.010 mmol, 0.5 equiv)
in DMF (0.5 mL). The resulting reaction mixture was stirred at rt
overnight, and purified by a Preparative reverse phase HPLC on a
Waters Autopurification system using a Sunfire Prep C18 OBD column
[5 .mu.m, 19.times.50 mm; flow rate, 20 mL/min; Solvent A: H.sub.2O
with 0.1% HCO.sub.2H; Solvent B: CH.sub.3CN with 0.1% HCO.sub.2H;
injection volume: 4.0 mL (CH.sub.3CN); gradient: 90.fwdarw.100% B
over 10 min; mass-directed fraction collection]. Fractions with the
desired MW, eluting at 5.4-6.6 min, were collected and concentrated
to give the desired amide 8-7-1 (11 mg, 66%): .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 16.01 (s, 1H), 9.53 (t, J=6.1 Hz, 1H), 7.83 (s,
1H), 7.50-7.48 (m, 2H), 7.40-7.32 (m, 8H), 5.35 (s, 2H), 4.87 (s,
2H), 3.97 (d, J=11.0 Hz, 1H), 3.90 (s, 3H), 3.39 (dd, J=4.9, 17.1
Hz, 1H), 3.13-3.04 (m, 2H), 3.00-2.93 (m, 1H), 2.58-2.54 (m, 1H),
2.49 (s, 6H), 2.47-2.42 (m, 1H), 2.36 (t, J=16.5 Hz, 1H), 2.17 (d,
J=14.6 Hz, 1H), 0.81 (s, 9H), 0.73 (s, 9H), 0.27 (s, 3H), 0.12 (s,
3H); MS (ESI) m/z 850.72 (M+H).
[0467] Aqueous HF (48-50%, 0.2 mL) was added to a solution of amide
(11 mg, 0.013 mmol, 1.0 equiv) in acetonitrile (0.5 mL) in a
polypropylene reaction vessel at 23.degree. C. The resulting
mixture was stirred vigorously at 23.degree. C. overnight and
poured into aqueous K.sub.2HPO.sub.4 (2.4 g dissolved in 25 mL
water). The resulting mixture was extracted with EtOAc (50 mL). The
organic phase was dried over anhydrous sodium sulfate, filtered,
and concentrated. The residue was used directly in the next step
without further purification.
[0468] The above crude product was dissolved in MeOH (1.5 mL) and
HCl/MeOH (0.5 N, 26 .mu.L, 1.0 equiv). Pd--C (10 wt %, 3.5 mg) was
added in one portion at 23.degree. C. The reaction vessel was
sealed and purged with hydrogen by briefly evacuating the flask
followed by flushing with hydrogen gas (1 atm). After stirring at
23.degree. C. for 30 min, the reaction mixture was filtered through
a small Celite pad. The filtrate was concentrated. The residue was
purified by a preparative reverse phase HPLC on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu.
RP-.gamma. 100A column [10 .mu.m, 150.times.21.20 mm; flow rate, 20
mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH.sub.3CN;
injection volume: 3.0 mL (0.05 N HCl/water); gradient:
25.fwdarw.60% B over 15 min; mass-directed fraction collection].
Fractions containing the desired product, eluting at 11.0-12.6 min,
were collected and freeze-dried to yield Compound 187 (3.0 mg, 39%
over 2 steps): .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.86 (s,
1H), 4.09 (s, 1H), 3.84 (s, 3H), 3.35-3.30 (m, 1H), 3.03-2.96 (m,
8H), 2.23-2.15 (m, 2H), 1.68-1.58 (m, 1H), 0.99 (s, 9H); MS (ESI)
m/z 558.38 (M+H).
[0469] Compound 188. Compound 188 was prepared Compound 187 using
dimethylaminoethylamine in place of neopentyl amine. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.89 (s, 1H), 4.11 (s, 1H), 3.85 (s,
3H), 3.85-3.82 (m, 2H), 3.41 (t, J=6.0 Hz, 2H), 3.37-3.32 (m, 1H),
3.04-2.96 (m, 14H), 2.23-2.15 (m, 2H), 1.68-1.58 (m, 1H); MS (ESI)
m/z 559.44 (M+H).
Example 9
Compounds of Formula II wherein Y is
--(CH.sub.2).sub.3--N(R.sup.2)(R.sup.3)
##STR00274##
[0471]
(4aS,11aR,12aS,13S)-3,7-bis(benzyloxy)-8-bromo-4a-(tert-butyldimeth-
ylsilyloxy)-13-(dimethylamino)-5-hydroxy-10-methoxy-11a,12,12a,13-tetrahyd-
rotetraceno[2,3-d]isoxazole-4,6(4aH,11H)-dione (9-1). n-BuLi (5.24
mL, 1.6 M/hexanes, 8.38 mmol, 2.0 equiv) was added dropwise to a
solution of diisopropylamine (1.18 mL, 8.38 mmol, 2.0 equiv) and
TMEDA (1.26 mL, 8.38 mmol, 2.0 equiv) in THF (40 mL) at -78 C. The
reaction solution was stirred at -78 C for 30 min. A solution of
ester 8-2 (1.79 g, 4.19 mmol, 1.0 equiv) in THF (20 mL) was added
via a cannula over 20 min. The resulting deep red solution was
stirred at -78 C for 20 min and was then cooled to -100 C. A
solution of enone 1-6 (1.62 g, 3.35 mmol, 0.8 equiv) in THF (20 mL)
was added to the reaction mixture via a cannula. The reaction
mixture was allowed to warm to -30 C over 1 h 30 min, quenched by a
mixture of saturated aqueous NH.sub.4Cl (100 mL). The resulting
mixture was extracted with EtOAc (200 mL, then 50 mL). The combined
EtOAc extracts were dried (sodium sulfate), filtered and
concentrated. The residue was purified by flash chromatography
(9:1:1 Hexanes/EtOAc/DCM) to give compound 9-1 (1.27 g, 46%):
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 16.00 (s, 1H), 7.59-7.57
(m, 2H), 7.52-7.50 (m, 2H), 7.41-7.32 (m, 6H), 7.26 (s, 1H), 5.37
(s, 2H), 5.00, 4.92 (ABq, J=9.2 Hz, 2H), 3.98 (d, J=10.4 Hz, 1H),
3.85 (s, 3H), 3.32 (dd, J=4.9, 15.9 Hz, 1H), 2.98-2.91 (m, 1H),
2.58-2.43 (m, 8H), 2.29 (t, J=15.9 Hz, 1H), 2.15 (d, J=14.0 Hz,
1H), 0.82 (s, 9H), 0.28 (s, 3H), 0.14 (s, 3H); MS (ESI) m/z 815.59,
817.59 (M+H).
[0472]
(4aS,11aR,12aS,13S)-3,7-bis(benzyloxy)-4a-(tert-butyldimethylsilylo-
xy)-8-((Z)-3-(tert-butyldimethylsilyloxy)allyl)-13-(dimethylamino)-5-hydro-
xy-10-methoxy-11a,12,12a,13-tetrahydrotetraceno[2,3-dfisoxazole-4,6(4aH,11-
H)-dione (9-3). A reaction vessel charged with bromide 9-1 (321 mg,
0.394 mmol, 1.0 equiv), Pd(OAc).sub.2 (17.7 mg, 0.079 mmol, 0.2
equiv) and P(o-Tol)3 (36 mg, 0.118 mmol, 0.3 eq) was vacuumed/back
flushed with N.sub.2 several times. Then DMF (2 mL), TEA (275
.mu.L, 1.97 mmol, 5.0 equiv) and allyl dimethyl-tert-butylsilyl
ether (168 .mu.L, 0.789 mmol, 2.0 equiv) were added under N2. The
reaction was then sealed and heated at 80.degree. C. for 6 h. The
reaction mixture was cooled to rt, diluted with MeCN, and purified
by a Preparative reverse phase HPLC on a Waters Autopurification
system using a Sunfire Prep C18 OBD column [5 .mu.m, 19.times.50
mm; flow rate, 20 mL/min; Solvent A: H.sub.2O with 0.1% HCO.sub.2H;
Solvent B: CH.sub.3CN with 0.1% HCO.sub.2H; injection volume: 4.0
mL (CH.sub.3CN); gradient: 90.fwdarw.100% B over 7 min;
mass-directed fraction collection]. Fractions with the desired MW
were collected and concentrated to afford the desired products 9-3
(containing small amount of the isomer) (212.9 mg, 60%): .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 16.07 (s, 1H), 7.42-7.34 (m, 4H),
7.31-7.22 (m, 7H), 6.87 (s, 1H), 6.23 (d, J=6.1 Hz, 1H), 5.27 (s,
2H), 4.75, 4.69 (ABq, J=10.0 Hz, 2H), 3.92 (d, J=10.4 Hz, 1H), 3.72
(s, 3H), 3.44-3.14 (m, 3H), 2.89-2.82 (m, 1H), 2.48-2.18 (m, 9H),
2.05 (d, J=14.6 Hz, 1H), 0.83 (s, 9H), 0.73 (s, 9H), 0.19 (s, 3H),
0.05 (s, 6H), 0.04 (s, 3H); MS (ESI) m/z 907.87 (M+H).
[0473]
3-((4aS,11aR,12aS,13S)-3,7-bis(benzyloxy)-4a-(tert-butyldimethylsil-
yloxy)-13-(dimethylamino)-5-hydroxy-10-methoxy-4,6-dioxo-4,4a,6,11,11a,12,-
12a,13-octahydrotetraceno[2,3-d]isoxazol-8-yl)propanal (9-4). Water
(0.5 mL) was added to a solution of 9-3 (162.4 mg, 0.179 mmol, 1.0
equiv). Then TFA (0.5 mL) was added dropwise at 0.degree. C. The
resulting reaction mixture was then stirred at rt for 2 h and
cooled to 0 C. Saturated sodium bicarbonate (8 mL) was added
slowly. The resulting mixture was then extracted with EtOAc (40 mL,
then 10 mL). The combined organic phase was dried over anhydrous
sodium sulfate, filtered and concentrated. The crude product 9-4
was used directly for the next step.
[0474] Compound 199.
##STR00275##
[0475] Pyrrolidine (19 .mu.L, 0.22 mmol, 5.0 equiv), acetic acid
(13 .mu.L, 0.22 mmol, 5.0 equiv) and sodium triacetoxyborohydride
(28 mg, 0.13 mmol, 3.0 equiv) were added sequentially to one fourth
of the above product 9-4 (0.045 mmol, 1.0 equiv) in
1,2-dichloroethane (1 mL) at 23.degree. C. After stirring for
overnight, the reaction mixture was quenched by the addition of
saturated aqueous sodium bicarbonate (5 mL) and pH 7 phosphate
buffer (10 mL) and extracted with dichloromethane (3.times.15 mL).
The combined organic extracts were dried over anhydrous sodium
sulfate, filtered, and concentrated. The residue was purified by a
Preparative reverse phase HPLC on a Waters Autopurification system
using a Sunfire Prep C18 OBD column [5 .mu.m, 19.times.50 mm; flow
rate, 20 mL/min; Solvent A: H.sub.2O with 0.1% HCO.sub.2H; Solvent
B: CH.sub.3CN with 0.1% HCO.sub.2H; injection volume: 4.0 mL
(CH.sub.3CN); gradient: 10.fwdarw.100% B over 10 min; mass-directed
fraction collection]. Fractions with the desired MW, eluting at
6.0-7.7 min, were collected and freeze-dried to afford the desired
products 9-5-1 (16 mg, 42% for 2 steps): .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 16.12 (br s, 1H), 7.50-7.48 (m, 2H), 7.44-7.42
(m, 2H), 7.39-7.30 (m, 6H), 6.90 (s, 1H), 5.35 (s, 2H), 4.80, 4.77
(ABq, J=10.4 Hz, 2H), 3.99 (d, J=10.4 Hz, 1H), 3.82 (s, 3H), 3.31
(dd, J=4.9, 15.9 Hz, 1H), 2.99-2.92 (m, 1H), 2.70-2.42 (m, 16H),
2.31 (t, J=15.3 Hz, 1H), 2.14 (d, J=14.6 Hz, 1H), 1.88-1.77 (m,
6H), 0.81 (s, 9H), 0.27 (s, 3H), 0.12 (s, 3H); MS (ESI) m/z 848.69
(M+H).
[0476] Aqueous HF (48-50%, 0.3 mL) was added to a solution of amine
9-5-1 (16 mg, 0.019 mmol, 1.0 equiv) in acetonitrile (0.6 mL) in a
polypropylene reaction vessel at 23.degree. C. The resulting
mixture was stirred vigorously at 23.degree. C. overnight and
poured into aqueous K.sub.2HPO.sub.4 (3.6 g dissolved in 25 mL
water). The resulting mixture was extracted with EtOAc (3.times.15
mL). The combined organic phase was dried over anhydrous sodium
sulfate, filtered, and concentrated. The residue was used directly
in the next step without further purification.
[0477] The above crude product was dissolved in MeOH (1.5 mL) and
HCl/MeOH (0.5 N, 76 .mu.L, 2.0 equiv). Pd--C (10 wt %, 4.3 mg) was
added in one portion at 23.degree. C. The reaction vessel was
sealed and purged with hydrogen by briefly evacuating the flask
followed by flushing with hydrogen gas (1 atm). After stirring at
23.degree. C. for 1 h, the reaction mixture was filtered through a
small Celite pad. The filtrate was concentrated. The residue was
purified by a preparative reverse phase HPLC on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu.
RP-.gamma. 100A column [10 .mu.m, 150.times.21.20 mm; flow rate, 20
mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH.sub.3CN;
injection volume: 3.0 mL (0.05 N HCl/water); gradient:
10.fwdarw.35% B over 15 min; mass-directed fraction collection].
Fractions containing the desired product, eluting at 11.6-13.5 min,
were collected and freeze-dried to yield Compound 199 (4.8 mg, 40%
over 2 steps): .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.19 (s,
1H), 4.08 (s, 1H), 3.80 (s, 3H), 3.69-3.63 (m, 2H), 3.28-3.21 (m,
3H), 3.08-2.93 (m, 10H), 2.77 (t, J=7.8 Hz, 2H), 2.22-2.00 (m, 8H),
1.65-1.55 (m, 1H); MS (ESI) m/z 556.51 (M+H).
[0478] Compounds 200-205 were prepared similarly to Compound 199,
using the appropriate amine NR.sup.2R.sup.3.
[0479] Compound 200. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.18
(s, 1H), 4.08 (s, 1H), 3.80 (s, 3H), 3.28-3.24 (m, 1H), 3.09-2.90
(m, 12H), 2.78 (t, J=7.3 Hz, 2H), 2.21-1.97 (m, 4H), 1.66-1.56 (m,
1H), 1.31 (t, J=7.3 Hz, 3H); MS (ESI) m/z 530.42 (M+H).
[0480] Compound 201. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.18
(s, 1H), 4.73 (dt, J=4.6, 47.2 Hz, 2H), 4.08 (s, 1H), 3.80 (s, 3H),
3.39 (dt, J=4.6, 26.6 Hz, 2H), 3.28-3.23 (m, 1H), 3.12-2.93 (m,
10H), 2.78 (t, J=7.8 Hz, 2H), 2.21-2.01 (m, 4H), 1.66-1.55 (m, 1H);
MS (ESI) m/z 548.47 (M+H).
[0481] Compound 202. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.18
(s, 1H), 6.30 (tt, J=2.8, 54.0 Hz, 1H), 4.08 (s, 1H), 3.80 (s, 3H),
3.56 (dt, J=3.2, 15.6 Hz, 2H), 3.28-3.23 (m, 1H), 3.17-3.13 (m,
2H), 3.04-2.93 (m, 8H), 2.78 (t, J=7.3 Hz, 2H), 2.22-2.02 (m, 4H),
1.65-1.55 (m, 1H); MS (ESI) m/z 566.49 (M+H).
[0482] Compound 203. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.18
(s, 1H), 4.08 (s, 1H), 4.04 (q, J=9.2 Hz, 1H), 3.80 (s, 3H),
3.28-3.23 (m, 1H), 3.21-3.17 (m, 2H), 3.04-2.93 (m, 8H), 2.78 (t,
J=7.3 Hz, 2H), 2.20-2.05 (m, 4H), 1.65-1.55 (m, 1H); MS (ESI) m/z
584.48 (M+H).
[0483] Compound 204. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.19
(s, 1H), 4.07 (s, 1H), 3.79 (s, 3H), 3.27-3.08 (m, 5H), 3.03-2.84
(m, 8H), 2.84 (s, 3H), 2.76 (t, J=7.3 Hz, 2H), 2.20-2.05 (m, 4H),
1.65-1.55 (m, 1H), 1.32 (t, J=7.3 Hz, 3H); MS (ESI) m/z 544.42
(M+H).
[0484] Compound 205. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.18
(s, 1H), 4.26-4.20 (m, 2H), 4.08-4.03 (m, 3H), 3.80 (s, 3H),
3.28-3.20 (m, 3H), 3.04-2.93 (m, 8H), 2.73 (t, J=7.3 Hz, 2H),
2.62-2.55 (m, 1H), 2.44-2.40 (m, 1H), 2.21-2.18 (m, 1H), 2.13-2.06
(m, 1H), 1.92-1.85 (m, 2H), 1.65-1.55 (m, 1H); MS (ESI) m/z 542.44
(M+H).
[0485]
(4aS,11aR,12aS,13S)-3,7-bis(benzyloxy)-4a-(tert-butyldimethylsilylo-
xy)-13-(dimethylamino)-5,8-dihydroxy-10-methoxy-11a,12,12a,13-tetrahydrote-
traceno[2,3-dfisoxazole-4,6(4aH,11H)-dione (9-7). 9-1 (46 mg, 0.056
mmol, 1.0 equiv) was dissolved in tetrahydrofurane (2 mL). Turbo
Grignard reagent (isopropylmagnesium chloride with lithium
chloride, 1.2 M, 113 .mu.L, 0.135 mmol) was added dropwise at
-78.degree. C. The mixture was warmed up to -20.degree. C. and
stirred for 1 h. Oxygen was then passed through the reaction
mixture slowly for 30 min. The reaction was then quenched by adding
5 mL ammonium chloride solution following addition of ethyl
acetate. The mixture was washed with H.sub.2O (10 mL) and
concentrated to give crude 9-7, which was purified by HPLC on a
Sunfire column to yield 4.0 mg yellow solid.
[0486] Compound 206. Aqueous HF (0.3 mL, 48-50%) was added to a
CH.sub.3CN solution (1.0 mL) of 9-7 in a plastic vial at 25 C. The
reaction was stirred at 25 C for 18 hrs. The resulting mixture was
poured into an aqueous solution (10 mL) of K.sub.2HPO.sub.4 (2 g).
The solution was extracted with EtOAc (3.times.15 mL). The combined
EtOAc extracts were dried over sodium sulfate and concentrated to
give the crude intermediate.
[0487] 10% Pd--C (20 mg) was added to a dioxane/MeOH solution (2
mL, 1:1) of the above crude intermediate. The reaction mixture was
stirred under H.sub.2 (balloon) at 25 C for 2 hrs and filtered
through a pad of Celite. The filtrate was concentrated to give the
crude product 144 mg. The crude product was purified by HPLC on a
Polymerx.TM. 10.mu., RP-.gamma. 100 R column [30.times.21.20 mm, 10
micron, solvent A: 0.05 N HCl, solvent B: CH.sub.3CN, sample in 2.0
mL (0.05 N HCl), gradient elution with 0.fwdarw.70% B over 15 min,
mass-directed fraction collection] to yield Compound 206 as a
yellow solid (1.64 mg, 6.3%, 3 steps): .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 6.82 (s, 1H), 4.05 (s, 1H), 3.74 (s, 3H), 3.17
(dd, J=15.6, 4.6 Hz, 1H), 3.02 (s, 3H), 2.96 (s, 3H), 3.01-2.89 (m,
2H), 2.19-2.11 (m, 1H), 2.09-2.00 (m, 1H), 1.65-1.53 (m, 1H); MS
(ESI) m/z 461.22 (M+H).
Example 10
Synthesis of Compounds of Formula III, wherein Y is
--NH--C(O)--CH.sub.2--NR.sup.2R.sup.3,
--NH--C(O)--(C.sub.1-C.sub.6)alkyl, --NH--C(O)-heterocyclyl, or
--NH--C(O)-carbocyclyl
##STR00276##
[0489] In Scheme 10, Z represents --(C.sub.1-C.sub.6)alkyl,
-heterocyclyl, or -carbocyclyl. Compounds 207-209 are compounds of
the invention, as well as being used as intermediates to produce
additional compounds of the invention.
[0490] Phenyl 2-methoxy-5-(trifluoromethoxy)benzoate (10-2). To
commerically availabe 2-methoxy-5-(trifluoromethoxy)benzoic acid
(10-1) (4.73 g, 18.91 mmol, 1 equiv) in dry DCM at room temperature
was added anhydrous DMF (5 drops) and oxalyl chloride (4.87 mL,
56.71 mmol, 3.0 equiv) dropwise. The reaction was stirred at room
temperature for 1 hr (until gas evolution ceased). The reaction
solution was concentrated in vacuo to yield the crude acid chloride
as a yellow oil, which was re-dissolved in dry DCM, and added with
PhOH (2.67 g, 28.37 mmol, 1.5 equiv), DIEA (9.88 mL, 56.72 mmol,
3.0 equiv) and DMAP (0.23 g, 1.88 mmol, 0.1 equiv). The reaction
was stirred at room temperature for overnight, quenched with
saturated sodium bicarbonate aqueous solution (100 mL), and
extracted with DCM (100 mL.times.3). The combined DCM extracts were
dried over sodium sulfate and concentrated in vacuo. Flash column
chromatography with 0% and 5% EtOAc/hexanes gave the desired phenyl
ester 10-2 as a pale oil (6.09 g, quantitative): R.sub.f 0.20 (10%
EtOAc/hexanes); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.87 (d,
J=2.4 Hz, 1H), 7.36-7.43 (m, 3H), 7.17-7.30 (m, 3H), 7.02 (d, J=9.2
Hz, 1H), 3.94 (s, 3H).
[0491] Phenyl 6-methoxy-2-methyl-3-(trifluoromethoxy)benzoate
(10-3). 2,2,6,6-Tetramethylpiperidine (TMP) (3.97 mL, 23.38 mmol,
1.2 equiv) was dissolved in anhydrous THF (50 mL) and cooled to
-78.degree. C. nBuLi (9.35 mL, 2.5 M in hexane, 23.38 mmol, 1.2
equiv) was added dropwise. The yellow solution was stirred at
0.degree. C. for 10 min and cooled to -78.degree. C. Anhydrous
N,N,N',N'-tetramethylethylenediamine (TMEDA) (4.38 mL, 29.22 mmol,
1.5 equiv) was added, followed by dropwise addition of phenyl ester
10-2 (6.09 g, 19.50 mmol, in 50 mL anhydrous THF, 1.0 equiv) over a
period of 15 min. The resulting deep yellow-brown solution was
stirred at -78.degree. C. for 30 min. Methyl iodide (1.82 mL, 29.23
mmol, 1.5 equiv) was added. The deep-brown solution was stirred
from -78.degree. C. to room temperature for 2 hrs and at room
temperature for 1 hr. HOAc (6 mL) and water (100 mL) were added
sequentially to quench the reaction. The bright orange solution was
concentrated in vacuo and extracted with EtOAc (200 mL.times.1).
The EtOAc extract was washed with 1 N aqueous HCl (50 mL.times.2)
and brine (50 mL.times.1), dried over sodium sulfate, and
concentrated in vacuo. Flash column chromatography with 0% and 5%
EtOAc/hexanes gave the desired phenyl ester 10-3 as a pale solid
(2.92 g, 46%): R.sub.f 0.30 (10% EtOAc/hexanes); .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.40-7.45 (m, 2H), 7.20-7.30 (m, 4H), 6.81
(d, J=8.5 Hz, 1H), 3.88 (s, 3H), 2.37 (s, 3H).
[0492] Phenyl 6-hydroxy-2-methyl-3-(trifluoromethoxy)benzoate
(10-4). Methyl ether 10-3 (500 mg, 1.53 mmol, 1 equiv) was
dissolved in anhydrous DCM (10 mL) and cooled to -78.degree. C.
BBr.sub.3 (1.60 mL, 1.0 M/DCM, 1.60 mmol, 1.02 equiv) was added
dropwise. The deep-orange solution was stirred from -78.degree. C.
to -10.degree. C. for 4 hrs. The reaction was quenched with
saturated sodium bicarbonate aqueous solution (100 mL) and
extracted with DCM (50 mL.times.3). The combined DCM extracts were
dried over sodium sulfate and concentrated in vacuo to yield the
crude phenol 10-4 as a yellow oil (410 mg, 86%): R.sub.f 0.55 (10%
EtOAc/hexanes); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 10.86 (s,
1H), 7.42-7.50 (m, 2H), 7.30-7.40 (m, 2H), 7.19 (d, J=7.9 Hz, 2H),
6.91 (d, J=9.2 Hz, 1H), 2.64 (s, 3H).
[0493] Phenyl
6-(tert-butoxycarbonyloxy)-2-methyl-3-(trifluoromethoxy)benzoate
(10-5). Crude phenol 10-4 (410 mg, 1.31 mmol, 1 equiv) was
dissolved in anhydrous DCM (10 mL). (Boc).sub.2O (430 mg, 1.97
mmol, 1.5 equiv), DIEA (0.46 mL, 2.64 mmol, 2 equiv), and DMAP (16
mg, 0.13 mmol, 0.1 equiv) were added sequentially at room
temperature. The reaction was stirred at room temperature for 1 hr,
quenched with saturated aqueous sodium bicarbonate (50 mL), and
extracted with DCM (50 mL.times.3). The combined DCM extracts were
dried over sodium sulfate and concentrated in vacuo. Flash column
chromatography with 0% and 2% EtOAc/hexanes gave the desired
product 10-5 as a white solid (540 mg, quantitative): R.sub.f 0.60
(10% EtOAc/hexanes); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.41-7.48 (m, 2H), 7.38 (d, J=10.3 Hz, 1H), 7.22-7.32 (m, 3H), 7.18
(d, J=9.2 Hz, 1H), 2.45 (s, 3H), 1.45 (s, 9H).
[0494]
(4aS,11aR,12aS,13S)-3-(benzyloxy)-4a-(tert-butyldimethylsilyloxy)-1-
3-(dimethylamino)-5-hydroxy-4,6-dioxo-10-(trifluoromethoxy)-4,4a,6,11,11a,-
12,12a,13-octahydrotetraceno[2,3-d]isoxazol-7-yl tert-butyl
carbonate (10-6). To diisopropylamine (0.062 mL, 0.44 mmol, 2.2
equiv) in THF (5 mL) at -78.degree. C. was added with nBuLi
dropwise (0.27 mL, 1.6 M/hexane, 0.44 mmol, 2.2 equiv). The pale
solution was brought to 0.degree. C., stirred at that temperature
for 10 min, and cooled to -78.degree. C. TMEDA (0.075 mL, 0.50
mmol, 2.5 equiv) was added, followed by dropwise addition of
compound 10-5 (91 mg, 0.22 mmol, 1.1 equiv, in 5 mL THF) over a
period of 10 min. The resulting deep-red solution was stirred at
-78.degree. C. for 15 min. Enone 1-6 (97 mg, 0.20 mmol, 1 equiv, in
5 mL THF) was added dropwise over a period of 2 min. The resulting
yellow solution was stirred from -78.degree. C. to -10.degree. C.
over a period of 1 hr, quenched with aqueous saturated ammonium
chloride (50 mL), and extracted with EtOAc (50 mL.times.3). The
extracts were combined, dried over sodium sulfate, and concentrated
in vacuo. Preparative HPLC purification on a C-18 column (mobile
phase: A-0.1% formic acid/water, B-0.1% formic acid/acetonitrile;
gradient: 80% B to 100% B over 10 min) yielded the desired product
10-6-1 as a yellow solid (90 mg, 56%): R.sub.f 0.40 (10%
EtOAc/hexane); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 15.48 (s,
1H), 7.36 (d, J=7.2 Hz, 2H), 7.29-7.20 (m, 4H), 6.94 (d, J=7.2 Hz,
1H), 5.22 (s, 2H), 3.81 (d, J=10.8 Hz, 1H), 3.15 (dd, J=16.0, 4.8
Hz, 1H), 2.94-2.85 (m, 1H), 2.44-2.31 (m, 9H), 2.03 (d, J=14.4 Hz,
1H), 1.42 (s, 9H), 0.78(s, 9H), 0.15(s, 3H), 0.08(s, 3H); MS (ESI)
m/z 801.3 (M+H).
[0495] (4aS,11aR,12aS,13S)-tert-butyl
3-(benzyloxy)-4a-(tert-butyldimethylsilyloxy)-13-(dimethylamino)-5,7-dihy-
droxy-4,6-dioxo-10-(trifluoromethoxy)-4,4a,6,11,11a,12,12a,13-octahydrotet-
raceno[2,3-dfisoxazole-8-carboxylate (10-6-2 The title compound
(10-6-2) was also isolated from the preparation of 10-6-1: MS (ESI)
m/z 801.3 (M+H).
[0496]
(4aS,11aR,12aS,13S)-3-(benzyloxy)-13-(dimethylamino)-4a,5,7-trihydr-
oxy-10-(trifluoromethoxy)-11a,12,12a,13-tetrahydrotetraceno[2,3-dfisoxazol-
e-4,6(4aH,11H)-dione (10-7). 10-6 (90 mg, 0.11 mmol) was dissolved
in acetonitrile (2 mL). HF (1 mL, 40% in water) was added. The
yellow solution was stirred at rt for overnight. The reaction
solution was then slowly added into K.sub.2HPO.sub.4 (4.3 g) in
water (20 mL) with rapid stirring. The mixture was extracted with
DCM (10 mL.times.3). The combined extracts were dried over sodium
sulfate and concentrated in vacuo to yield the crude product 10-7
as a yellow solid (76 mg).
[0497]
(4S,4aS,5aR,12aS)-4-(dimethylamino)-3,10,12,12a-tetrahydroxy-1,11-d-
ioxo-7-(trifluoromethoxy)-1,4,4a,5,5a,6,11,12a-octahydrotetracene-2-carbox-
amide (Compound 207). 10-7 (crude, 0.11 mmol, 1 equiv) was
dissolved 1,4-dioxane/methanol (5 mL, 1:4 v/v). 10% Pd--C (47 mg,
0.022 mmol Pd, 0.2 equiv) was added. The mixture was purged by
bubbling hydrogen through for 5 min and rapidly stirred under 1 atm
hydrogen atmosphere at rt for 1 hr. The catalyst was filtered off
with a small Celite pad and washed with more methanol (2
mL.times.3). The filtrate was concentrated in vacuo. Preparative
HPLC purification on a Polymerx.TM. column yielded the desired
product Compound 207 as a yellow solid (72 mg, quantitative):
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.45 (dd, J=9.2, 0.9 Hz,
1H), 6.91 (d, J=9.2 Hz, 1H), 4.11 (s, 1H), 3.19-2.96 (m, 9H),
2.36-2.30 (m 1H), 2.25-2.20 (m, 1H), 1.69-1.63 (m, 1H); MS (ESI)
m/z 499.4 (M+H).
[0498]
(4S,4aS,5aR,12aS)-4-(dimethylamino)-3,10,12,12a-tetrahydroxy-9-nitr-
o-1,11-dioxo-7-(trifluoromethoxy)-1,4,4a,5,5a,6,11,12a-octahydrotetracene--
2-carboxamide (Compound 208). Compound 207 (70 mg, 0.14 mmol, 1
equiv) was dissolved in cold (0.degree.) sulfuric acid (4 mL). A
solution of HNO.sub.3 in H.sub.2SO.sub.4 (0.42 mL, 0.5 M, 0.21
mmol, prepared prior to use by mixing 70% aqueous nitric acid with
concentrated sulfuric acid) was added dropwise at 0.degree. C. The
deep-red solution was stirred at 0.degree. C. for 1 hr. The
resulting yellow solution was added dropwise into diethyl ether
(200 mL) with rapid stirring. The yellow solid was collected,
washed with more diethyl ether (5 mL.times.4), and dried in vacuo
to yield the crude product Compound 208 as a yellow solid, which
was used directly in the next step.
[0499]
(4S,4aS,5aR,12aS)-9-amino-4-(dimethylamino)-3,10,12,12a-tetrahydrox-
y-1,11-dioxo-7-(trifluoromethoxy)-1,4,4a,5,5a,6,11,12a-octahydrotetracene--
2-carboxamide (Compound 209). Compound 208 (0.10 mmol, crude) was
dissolved in 0.1 N HCl/methanol (5 mL). 10% Pd--C (54 mg, 0.02 mmol
Pd, 0.2 equiv) was added. The mixture was purged by bubbling
hydrogen through for 5 min and rapidly stirred under 1 atm hydrogen
atmosphere at rt for 1 hr. The catalyst was filtered off with a
small Celite pad and washed with more methanol (5 mL.times.4). The
filtrate was concentrated in vacuo. Preparative HPLC purification
on a Polymerx.TM. column yielded the desired aniline Compound 209
as a brown solid (13 mg, 22%): .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.60 (s, 1H), 4.12 (s, 1H), 3.21-2.93 (m, 9H), 2.38-2.32
(m, 1H), 2.25-2.22 (m, 1H), 1.69-1.59 (m, 1H); MS (ESI) m/z
514.1(M+H).
[0500] Compound 210.
[0501] Compound 209 (4 mg, 0.0068 mmol, 1 equiv) was dissolved in
acetonitrile/DMPU (1 mL, 1:3 v/v). Sodium carbonate (4 mg, 0.04
mmol, 6 equiv) and bromoacetyl bromide/acetonitrile (0.041 mL, 0.2
M, 0.0082 mmol, 1.2 equiv, prepared prior to use by dissolving
bromoacetyl bromide in anhydrous acetonitrile) was added at rt. The
reaction was stirred at rt for 10 min. LC/MS analysis indicated
complete consumption of the starting aniline 6. Azetidine (0.0092
mL, 0.14 mmol, 20 equiv) was added. The reaction was stirred at rt
for 1 hr. LC/MS analysis indicated complete consumption of the
intermediate bromide. The reaction mixture was then added dropwise
into 5 mM HCl/diethyl ether (50 mL) with rapid stirring. The solid
was collected on a small Celite pad, washed with more 5 mM
HCl/diethyl ether (5 mL.times.4), and eluted with 0.1 N HCl/water
(4 mL). The yellow eluent was then directly injected into a
preparative HPLC system equipped with a Polymerx.TM. column for
purification. The desired product Compound 210 was obtained as a
yellow solid after freeze-drying (3.2 mg, 2 HCl, 69%): .sup.1H NMR
(400 MHz, CD.sub.3OD) .delta. 8.40 (s, 1H), 4.30-4.40 (m, 4H),
4.10-4.23 (m, 2H), 4.09 (s, 1H), 2.90-3.50 (m, 3H), 3.03 (s, 3H),
2.94 (s, 3H), 2.58-2.70 (m, 1H), 2.42-2.52 (m, 1H), 2.28-2.36 (m,
1H), 2.18-2.25 (m, 1H), 1.58-1.70 (m, 1H); MS (ESI) m/z 611.3
(M+H).
[0502] The following compounds were prepared similarly to Compound
210 substituting the appropriate amine (NHR.sup.2R.sup.3).
[0503] Compound 211. .sup.1HNMR (400 MHz, CD.sub.3OD): .delta. 8.45
(s, 1H), 4.23 (s, 2H), 4.11-4.09 (m, 3H), 3.19-2.93 (m, 9H),
2.35-2.26 (m, 1H), 2.25-2.17 (m, 1H), 1.68-157 (m, 1H); MS (ESI)
m/z 653.1(M+H).
[0504] Compound 212. .sup.1 HNMR (400 MHz, CD.sub.3OD): .delta.
8.39 (s, 1H), 5.53-5.35 (m, 1H), 4.58-4.42 (m, 6H), 4.10 (s, 1H),
3.15-2.93 (m, 9H), 2.35-2.25 (m, 1H), 2.23-2.18 (m, 1H), 1.37-158
(m, 1H); MS (ESI) m/z 629.1(M+H).
[0505] Compound 213. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.43
(s, 1H) , 4.11-4.10 (m, 3H), 3.18-2.96 (m,11H), 2.36-2.21 (m, 2H),
1.75-1.63 (m, 3H), 1.49-1.43 (m, 2H), 1.02 (t, J=7.2 Hz, 3H); MS
(ESI) m/z 627.2 (M+H).
[0506] Compound 214. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.47
(s, 1 H, 4.14 (s, 3H), 3.22-3.00 (m,11H), 2.39-2.26 (m, 2H),
1.79-1.77 (m, 2H), 1.69-1.67 (m, 1H), 1.46-1.45 (m, 4H), 1.02 (t,
J=7.2 Hz, 3H); MS (ESI) m/z 641.2(M+H).
[0507] Compound 215. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.46
(s, 1H), 4.12 (s, 3H), 3.21-2.98 (m,11H), 2.38-2.31 (m, 2H),
1.79-1.75 (m, 2H), 1.69-1.66 (m, 1H), 1.48-1.39 (m, 6H), 0.98 (t,
J=7.2 Hz, 3H); MS (ESI) m/z 655.2(M+H).
[0508] Compound 216. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.43
(s, 1H) , 4.10(s, 2H), 4.08 (s, 1H), 3.17-2.94 (m,11H), 2.35-2.27
(m, 1H), 2.25-2.18 (m, 1H), 1.64-1.61 (m, 1H), 1.10 (m, 9H); MS
(ESI) m/z 641.1(M+H).
[0509] Compound 217. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.34
(s, 1H), 4.04 (s, 2H), 4.01 (s, 1H), 3.11-2.87 (m,11H), 2.26-2.13
(m, 2H), 1.60-1.53 (m, 1H), 1.18-1.05 (m, 1H), 0.67-0.63 (m, 2H),
0.37-0.33 (m, 2H); MS (ESI) m/z 625.2(M+H).
[0510] Compound 218. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.44
(s, 1H), 4.35-4.20 (m, 2H), 4.13 (s, 1H), 3.43-3.33 (q, J=7.2 Hz,
2H), 3.22-2.98 (m, 12H), 2.37-2.24 (m, 2H), 2.22-2.09 (m, 1H),
1.45-1.42 (t, J=7.2 Hz, 3H); MS (ESI) m/z 613.2(M+H).
[0511] Compound 219. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.49
(s, 1H), 7.44 (s, 4H), 5.06 (brs, 2H), 4.73 (brs, 2H), 4.63 (s,
2H), 4.14 (s, 1H), 3.21-2.98 (m, 9H), 2.38-2.27 (m, 2H), 1.70-1.67
(m, 1H); MS (ESI) m/z 673.2(M+H).
[0512] Compound 220. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.44
(s, 1H), 4.10 (s, 3H), 3.50-3.47(m, 1H), 3.18-2.96 (m, 9H),
2.36-2.22 (m, 2H), 1.65-1.63 (m, 1H), 1.38 (t, J=6.8 Hz, 6H); MS
(ESI) m/z 613.2(M+H).
[0513] Compound 221. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.44
(s, 1H), 4.73-4.68 (m, 1H), 4.45-4.37 (m, 5H), 4.15 (s, 1H),
4.14-4.10 (m, 1H), 3.40 (s, 3H), 3.22-2.92 (m, 9H), 2.40-2.32 (m,
2H), 1.70-1.67 (m, 1H); MS (ESI) m/z 641.3(M+H).
[0514] Compound 222. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.35
(s, 1H), 4.81 (t, J=4.4 Hz, 1H), 4.64 (t, J=4.4 Hz, 1H), 4.09 (s,
2H), 4.00 (s, 1H), 3.45 (t, J=4.4 Hz, 1H), 3.39 (t, J=4.4 Hz, 1H),
3.09-2.79 (m, 9H), 2.27-2.09 (m, 2H), 1.62-1.58(m, 1H); MS (ESI)
m/z 617.2(M+H).
[0515] Compound 223. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.46
(s, 1H), 4.12 (s, 3H), 3.32-2.97 (m, 11H), 2.38-2.27 (m, 2H),
1.99-1.90 (s, 3H), 1.74-1.64 (m, 5H), 1.33 (m, 2H); MS (ESI) m/z
653.1(M+H).
[0516] Compound 224. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.34
(s, 1H), 4.02 (s, 1H), 4.00 (s, 2H), 3.10 (q, J=7.2 Hz, 2H),
3.10-2.87 (m, 9H), 2.28-2.15 (m, 2H),1.57-1.46 (m, 1H), 1.27 (t,
J=7.2 Hz, 3H); MS (ESI) m/z 599.1(M+H).
[0517] Compound 225. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.43
(s, 1H), 4.35-4.20 (m, 2H), 4.11 (s, 1H), 3.38-2.94 (m, 14H),
2.35-2.21(m, 2H), 1.89-1.79 (m, 2H), 1.68-1.63 (m, 1H), 1.09 (t,
J=7.2 Hz, 3H); MS (ESI) m/z 627.1 (M+H).
[0518] Compound 226. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.35
(s, 1H), 4.04 (s, 2H), 4.00 (s, 1H), 3.61 (t, J=4.8 Hz, 2H), 3.38
(s, 3H), 3.21 (t, J=4.8 Hz, 2H), 3.15-2.86 (m, 9H), 2.18-2.09 (m,
2H), 1.61-1.52 (m, 1H); MS (ESI) m/z 629.1(M+H).
[0519] Compound 227. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.34
(s, 1H), 5.47-5.32 (m, 1H), 4.34 (s, 2H), 4.02 (s, 1H), 3.98-3.82
(m, 2H), 3.3.48-3.32 (m, 2H), 3.11-2.87 (m, 9H), 2.47-2.36 (m, 2H),
2.34-2.19 (m, 2H), 1.58-1.50(m, 1H); MS (ESI) m/z 643.1(M+H).
[0520] Compound 228. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.30
(s, 1H), 8.06 (s, 1H), 7.94 (d, J=4.8 Hz, 1H), 7.73-7.67 (m, 2H),
4.17 (s, 2H), 4.02 (s, 1H), 3.24-2.86 (m, 9H), 2.25-2.12 (m, 2H),
1.59-1.49(m, 1H); MS (ESI) m/z 648.1(M+H).
[0521] Compound 229. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.35
(s, 1H), 8.02 (d, J=7.2 Hz, 2H), 6.84 (d, J=7.2 Hz, 2H), 5.17 (s,
2H),4.09 (s, 1H), 3.28-2.936 (m, 9H), 2.32-2.19 (m, 2H),
1.66-1.56(m, 1H); MS (ESI) m/z 648.1(M+H).
[0522] Compound 230. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.43
(s, 1H), 4.07 (s, 3H), 3.20-2.94 (m, 11H), 2.35-2.29 (m, 1H),
2.25-2.17 (m, 1H),1.93-1.79 (m, 2H), 1.78-1.58 (m, 1H), 1.05 (t,
J=7.2 Hz, 3H); MS (ESI) m/z 613.2 (M+H).
[0523] Compound 231. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.42
(s, 1H), 4.12 (s, 3H), 3.19-2.87 (m, 11H), 2.38-2.19 (m, 2H),
1.90-1.61 (m, 6H), 1.38-1.19 (m, 4H), 1.15-1.01 (m, 2H); MS (ESI)
m/z 667.2 (M+H).
[0524] Compound 232. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.35
(s, 1H), 4.01 (s, 3H), 3.14-2.73 (m, 11H), 2.26-2.13 (m, 2H),
2.03-2.01 (m, 1H), 1.60-1.51 (m, 1H), 0.99-0.92 (t, J=6.8 Hz, 6H);
MS (ESI) m/z 627.2 (M+H).
[0525] Compound 233. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.34
(s, 1H), 4.12 (s, 2H), 4.01 (s, 1H), 3.56-3.53 (m, 2H), 3.11-2.73
(m, 11H), 2.27-2.13 (m, 2H), 1.89-1.76 (m, 5H), 1.60-1.57 (m, 2H);
MS (ESI) m/z 639.2 (M+H).
[0526] Compound 234. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.35
(s, 1H), 4.09 (s, 1H), 4.02 (s, 2H), 3.89-3.70 (m, 1H), 3.11-2.86
(m, 9H), 2.27-2.02 (m, 6H), 1.91-1.71 (m, 2H), 1.60-1.40 (m, 1H);
MS (ESI) m/z 625.3 (M+H).
[0527] Compound 235. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.44
(s, 1H), 4.31 (s, 2H), 4.12 (s, 1H), 4.12-3.78 (m, 4H), 3.78-3.44
(m, 4H), 3.22-2.90 (m, 9H), 2.38-2.23 (m, 2H), 1.71-1.61 (m, 1H);
MS (ESI) m/z 641.1 (M+H).
[0528] Compound 236. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.42
(s, 1H), 4.27 (s, 2H), 4.11 (s, 1H), 3.40-3.31 (m, 4H), 3.21-2.96
(m, 9H), 2.36-2.23 (m, 2H), 1.69-1.60 (m, 1H), 1.40 (t, J=7.2 Hz,
6H); MS (ESI) m/z 627.2 (M+H).
[0529] Compound 237. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.46
(s, 1H), 4.11 (s, 3H), 3.31-2.94 (m, 10H), 2.40-2.20 (m, 2H),
2.20-2.00 (m, 2H), 1.94-1.82 (m, 2H), 1.68-1.56 (m, 1H), 1.48-1.32
(m, 4H), 1.28-1.16 (m, 2H); MS (ESI) m/z 653.3 (M+H).
[0530] Compound 238. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.41
(s, 1H), 7.46-7.26 (m, 2H), 7.25-7.11 (m, 3H), 4.16 (s, 2H), 4.01
(s, 1H), 3.15-2.78 (m, 9H), 2.26-2.11 (m, 2H), 1.58-1.49 (m, 1H);
MS (ESI) m/z 647.0 (M+H).
[0531] Compound 239. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.43
(s, 1H), 4.21 (s, 2H), 4.11 (s, 1H), 3.18-2.84 (m, 9H), 2.90-2.84
(m, 1H), 2.35-2.22 (m, 2H), 1.69-1.59 (m, 1H), 0.95-0.93 (m, 4H);
MS (ESI) m/z 611.4(M+H).
[0532] Compound 240. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.43
(s, 1H), 6.38 (tt, J=60, 1.6 Hz, 1H), 4.21 (s, 2H), 4.12 (s, 1H),
3.68 (td, J=7.2, 1.6 Hz, 2H), 3.18-2.90 (m, 9H), 2.36-2.18 (m, 2H),
1.69-1.58 (m, 1H); MS (ESI) m/z 635.1 (M+H).
[0533] Compound 241. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.46
(s, 1H), 5.56-5.43 (m, 1H), 4.45 (s, 2H), 4.12 (s, 1H), 4.13-3.90
(m, 2H), 3.58-3.41 (m, 2H), 3.20-2.90 (m, 9H), 2.38-2.23 (m, 4H),
1.71-1.61 (m, 1H); MS (ESI) m/z 643.2 (M+H).
[0534] Compound 242. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
8.35-8.33 (m, 1H), 4.22-4.18 (m, 2H), 4.02 (s, 1H), 3.92-3.87 (m,
1H), 3.39-3.28 (m, 1H), 3.09-2.86 (m, 11H), 2.74-2.60 (m, 2H),
2.27-2.12 (m, 2H), 1.71-1.45 (m, 7H); MS (ESI) m/z 665.2 (M+H).
[0535] Compound 243. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.44
(s, 1H), 4.12 (s, 1H), 4.08 (s, 2H), 3.18-2.90 (m, 11H), 2.78-2.66
(m, 1H), 2.38-2.15 (m, 4H), 2.08-1.80 (m, 3H), 1.71-1.54 (m, 1H);
MS (ESI) m/z 639.2 (M+H).
[0536] Compound 244. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.38
(s, 1H), 4.20 (s, 2H), 4.02 (s, 1H), 3.18-2.82 (m, 15H), 2.32-2.11
(m, 2H), 1.66-1.52 (m, 1H); MS (ESI) m/z 599.1 (M+H).
[0537] Compound 245. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.46
(s, 1H), 4.09 (s, 3H), 2.90-3.30 (m, 3H), 3.03 (s, 3H), 2.94 (s,
3H), 2.29-2.37 (m, 1H), 2.18-2.25 (m,1H), 1.58-1.70 (m, 1H), 1.41
(s, 9H); MS (ESI) m/z 627.4 (M+H).
[0538] Compound 246. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.44
(s, 1H), 4.33 (s, 2H), 4.10 (s, 1H), 3.75-3.85 (m, 2H), 2.90-3.50
(m, 7H), 3.03 (s, 3H), 2.94 (s, 3H), 2.28-2.37 (m, 1H), 2.00-2.25
(m, 5H), 1.58-1.70 (m, 1H); MS (ESI) m/z 625.1 (M+H).
[0539] Compound 263. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.46
(s, 1H), 4.12 (s, 1H), 4.11 (s, 2H), 3.70-3.61 (m, 1H), 3.22-2.93
(m, 9H), 2.38-2.10 (m, 4H), 1.92-1.80 (m, 2H), 1.76-1.56 (m, 5H);
MS (ESI) m/z 639.2 (M+H).
[0540] The following compounds were prepared similarly to Compound
210 substituting bromoacetyl bromide and an amine
(NHR.sup.2R.sup.3) with an acid chloride (appropriately protected
if needed) or a carboxylic acid (appropriately protected if needed)
and a coupling reagent such as HATU.
[0541] Compound 247. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.34
(s, 1H), 7.88 (d, J=7.6 Hz, 2H), 7.55-7.45 (m, 3H), 4.03 (s, 1H),
3.13-2.88 (m, 9H), 2.33-2.28 (m, 1H), 2.19-2.13 (m, 1H), 1.64-1.53
(m, 1H); MS (ESI) m/z 611.4(M+H).
[0542] Compound 248. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.37
(s, 1H), 7.50-7.41 (m, 3H), 7.16 (dd, J=8.0 Hz, 1.6 Hz, 1H), 4.09
(s, 1H), 3.86 (s, 3H),3.18-2.97 (m, 9H), 2.37-2.29 (m, 1H),
2.24-2.20 (m, 1H), 1.69-1.62 (m, 1H); MS (ESI) m/z 648.2(M+H).
[0543] Compound 249. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.24
(s, 1H), 8.16 (s, 1H), 8.12 (d, J=8.0 Hz, 1H), 7.83 (d, J=8.0 Hz,
1H), 7.66 (t, J=8.0 Hz, 1H), 4.01 (s, 1H), 3.12-2.73 (m, 9H), 2.26
(m, 1H), 2.16-2.12 (m, 1H), 1.63-1.52 (m, 1H); MS (ESI) m/z
686.1(M+H).
[0544] Compound 250. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.29
(s, 1H), 7.88 (d, J=3.6 Hz, 1H), 7.77 (d, J=4.8 Hz, 1H), 7.21 (t,
J=4.4 Hz, 1H), 4.10 (s, 1H), 3.19-2.96 (m, 9H), 2.38-2.30 (m, 1H),
2.28-2.20 (m, 1H), 1.70-1.60 (m, 1H); MS (ESI) m/z 624.1(M+H).
[0545] Compound 251. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.29
(s, 1H), 7.98 (d, J=8.4 Hz, 2H), 7.44 (d, J=8.8 Hz, 2H), 4.03 (s,
1H), 3.18 (s, 6H), 3.11-2.87 (m, 9H), 2.25-2.20 (m, 1H), 2.16-2.13
(m, 1H), 1.58-1.55 (m, 1H); MS (ESI) m/z 661.1(M+H).
[0546] Compound 252. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.25
(s, 1H), 8.17 (s, 1H), 8.04 (d, J=7.6 Hz, 1H), 7.85 (dd, J=7.2 Hz,
1.6 Hz,1H), 7.70 (t, J=8.0 Hz, 1H), 4.04 (s, 1H), 3.20 (s, 6H),
3.12-2.88 (m, 9H), 2.26 (m, 1H), 2.18-2.15 (m, 1H), 1.58-1.55 (m,
1H); MS (ESI) m/z 661.1(M+H).
[0547] Compound 253. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.26
(s, 1H), 8.10 (d, J=7.6 Hz, 1H), 7.95 (d, J=8.0 Hz, 1H), 7.80 (t,
J=7.6Hz, 1H), 7.65 (t, J=7.6 Hz, 1H), 4.03 (s, 1H), 3.26 (s, 6H),
3.14-2.87 (m, 9H), 2.29-2.25 (m, 1H), 2.13-2.10 (m, 1H), 1.63-1.56
(m, 1H); MS (ESI) m/z 661.1(M+H).
[0548] Compound 254. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.38
(s, 1H), 9.08-9.04 (m, 2H), 8.37 (s, 1H), 8.22 (t, J=7.6 Hz, 1H),
4.13 (s, 1H), 3.22-2.98 (m, 9H), 2.36 (m, 1H), 2.13-2.10 (m, 1H),
1.70-1.63 (m, 1H); MS (ESI) m/z 619.0(M+H).
[0549] Compound 255. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 9.07
(d, J=5.6 Hz, 2H), 8.49 (d, J=5.6 Hz, 2H), 8.37 (s, 1H), 4.12 (s,
1H), 3.22-2.97 (m, 9H), 2.37 (m, 1H), 2.29-2.23 (m, 1H), 1.70-1.63
(m, 1H); MS (ESI) m/z 619.0(M+H).
[0550] Compound 256. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
8.72-8.70 (m, 2H), 8.23 (d, J=7.2 Hz, 1H), 8.06 (t, J=7.6 Hz,1H),
7.65 (dd, J=10.8, 6.8 Hz, 1H), 4.12 (s, 1H), 3.19-2.97 (m, 9H),
2.32-2.25 (m, 1H), 2.26-2.23 (m, 1H), 1.67-1.64 (m, 1H); MS (ESI)
m/z 619.0(M+H).
[0551] Compound 257. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33
(s, 1H), 4.09-4.08 (m, 2H), 3.45-3.42 (m, 1H), 3.16-2.93 (m, 10H),
2.34-2.18 (m, 3H), 1.95-1.88 (m, 2H), 1.88-1.60 (m, 4H); MS (ESI)
m/z 625.1 (M+H)
[0552] Compound 258. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.44
(s, 1H), 5.20 (t, J=9.2 Hz, 1H), 4.19-4.07 (m, 3H), 3.19-2.92 (m,
12H), 2.66-2.61 (m, 2H), 2.33 (m, 1H), 2.22-2.03 (m, 1H),1.66-1.63
(m, 1H); MS (ESI) m/z 611.1 (M+H)
[0553] Compound 259. .sup.iH NMR (400 MHz, CD.sub.3OD) .delta. 8.34
(s, 1H), 4.39 (t, J=7.6 Hz, 1H), 4.10 (s, 1H), 3.76-3.74 (m, 1H),
3.27-2.93 (m, 12H), 2.70-2.63 (m, 1H), 2.34-2.15 (m, 6H), 1.63-1.60
(m, 1H); MS (ESI) m/z 625.1(M+H).
[0554] Compound 260. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.36
(s, 1H), 5.20 (t, J=8.0 Hz, 1H), 4.09-4.07 (m, 1H), 4.02 (s, 1H),
3.96-3.73 (m, 1H), 3.21-2.72 (m, 11H), 2.59-2.13 (m, 2H), 1.56-1.54
(m, 1H); MS (ESI) m/z 597.1 (M+H).
[0555] Compound 261. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33
(s, 1H), 4.57 (t, J=7.6 Hz, 1H), 4.09 (s, 1H), 3.47-3.36 (m, 2H),
3.18-2.93 (m, 9H), 2.54-2.52 (m, 1H), 2.34-2.31 (m,1H),2.27-2.07
(m, 4H),1.63-1.58 (m,1H); MS (ESI) m/z 611.1 (M+H).
[0556] Compound 262. Compound 262 was prepared from 10-6-2 via
aqueous HF treatment followed by hydrogenation similarly to
Compound 210: .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.00 (s,
1H), 4.09 (s, 1H), 2.80-3.50 (m, 9H), 2.32-2.42 (m, 1H), 2.19-2.27
(m, 1H), 1.58-1.70 (m, 1H); MS (ESI) m/z 543.3 (M+H).
Example 11
Synthesis of Compounds of Formula III, wherein Y is
NR.sup.2R.sup.3
##STR00277##
[0558] In Scheme 11, "R.sup.D" represents C.sub.1-C.sub.6 alkyl;
and "R.sup.E" represents C.sub.1-C.sub.6 alkyl or phenyl.
Phenyl 2-hydroxy-6-methyl-3-nitro-5-(trifluoromethoxy)benzoate
(11-1)
[0559] Compound 10-4 (12.2 mmol) was dissolved in
1,2-dichloroethane (18 mL) and H.sub.2O (18 mL). Tetrabutylammonium
bromide (197 mg, 0.61 mmol, 0.05 eq) was added, followed by slow
addition of nitric acid (70%, 1.56 mL, 24.4 mmol, 2.0 eq) in order
to maintain temperature below 25.degree. C. After stirred at rt for
20 h, the reaction mixture was diluted with dichloromethane and
layers were separated. The organic layer was further washed with
H.sub.2O, brine, dried (Na.sub.2SO.sub.4) and concentrated to give
compound 11-1 as yellow solid, which was pure enough to use
directly for the next step.
[0560] Phenyl
2-(benzyloxy)-6-methyl-3-nitro-5-(trifluoromethoxy)benzoate (11-2).
11-1 obtained from the above (12.2 mmol) was dissolved in acetone
(25 mL). Potassium iodide (101 mg, 0.61 mmol, 0.05 eq),
K.sub.2CO.sub.3 (3.37 g, 24.4 mmol, 2 eq), and benzylbromide (1.74
mL, 14.6 mmol, 1.2 eq) were added. The resulting mixture was heated
to reflux for 4 h. After cooled to rt, the solution was filtered
through a bed of Celite. The solid cake was further washed with
three portions of EtOAc. The combined organic solution was
concentrated. The residue was dissolved in EtOAc and washed with
H.sub.2O, brine, dried (Na.sub.2SO.sub.4) and concentrated to give
compound 11-2 as off-white solid, which was used directly for the
next step without further purification.
[0561] Phenyl
3-amino-2-(benzyloxy)-6-methyl-5-(trifluoromethoxy)benzoate (11-3).
11-2 prepared from the last step (12.2 mmol) was dissolve in THF
(73 mL). Solution of Na.sub.2S.sub.2O.sub.4 (12.49 g, 61.0 mmol, 5
equiv) in H.sub.2O (49 mL) was added at 0.degree. C. After stirred
at rt for 16 h, layers were separated. The aqueous layer was
further extracted with EtOAc three times. The combined organic
layers were washed with brine, dried (Na.sub.2SO.sub.4) and
concentrated. Further purification with flash chromatography
(silica gel, 85:15 hexanes/EtOAc) yielded compound 11-3 as
off-white solid (3.45 g): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.44-7.34 (m, 7H), 7.28-7.25 (m, 1H), 7.14-7.12 (m, 2H), 6.73 (s,
1H), 5.01 (s, 2H), 3.86 (br s, 2H), 2.29 (s, 3H); MS (ESI) m/z
416.3 (M-H), calcd for C.sub.22H.sub.17F.sub.3NO.sub.4 416.12.
[0562] Phenyl
2-(benzyloxy)-3-(bis(tert-butoxycarbonyl)amino)-6-methyl-5-(trifluorometh-
oxy)benzoate (11-4). Di-tert-butyl dicarbonate (4.51 g, 20.66 mmol,
2.5 eq) and DMAP (50 mg, 0.41 mmol, 0.05 eq) were added to the
solution of compound 11-3 (3.45 g, 8.26 mmol, 1 eq) in anhydrous
DMF (33 mL). The resulting mixture was stirred at rt and the
reaction was monitored by LC-MS. After SM was completely consumed,
the reaction was diluted with EtOAc. The solution was washed with
H.sub.2O three times, brine, dried over Na.sub.2SO.sub.4, filtered
and concentrated. Further purification with flash chromatography
(silica gel, 95:5 hexanes/EtOAc) yielded compound 11-4 as wax-like
solid (4.12 g):.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.40-7.31
(m, 7H), 7.28-7.25 (m, 1H), 7.15 (s, 1H), 6.98 (d, J=7.8 Hz, 1H),
4.94 (s, 2H), 2.40 (s, 3H), 1.37 (s, 18H); MS (ESI) m/z 616.4
(M-H), calcd for C.sub.32H.sub.33F.sub.3NO.sub.8 616.22.
[0563] Intermediate 11-5. 11-5 was prepared from compound 11-4 and
eone 1-6 under similar conditions used in the preparation of
compound 2-5 (see Example 2). The crude product was purified by
flash chromatography (silica gel, 95:5 to 85:15 hexanes/EtOAc) gave
desired product as light yellow foam: .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 15.88 (s, 1H), 7.52-7.30 (m, 11H), 5.35 (s,
2H), 4.98 (d, J=9.8 Hz, 1H), 4.79 (d, J=9.8 Hz, 1H), 3.96 (d,
J=10.4 Hz, 1H), 3.26 (dd, J=16.5, 4.9 Hz, 1H), 3.01-2.93 (m, 1H),
2.60-2.40 (m, 9H), 2.16-2.13 (m, 1H), 1.34 (s, 18H), 0.81 (s, 9H),
0.26 (s, 3H), 0.13 (s, 3H); MS (ESI) m/z 1006.7 (M+H), calcd for
C.sub.52H.sub.63F.sub.3N.sub.3O.sub.12Si 1006.41.
[0564]
(4aS,11aR,12aS,13S)-8-amino-3,7-bis(benzyloxy)-4a-(tert-butyldimeth-
ylsilyloxy)-13-(dimethylamino)-5-hydroxy-10-(trifluoromethoxy)-11a,12,12a,-
13-tetrahydrotetraceno[2,3-d]isoxazole-4,6(4aH,11H)-dione (11-6).
11-6 was prepared from 11-5 under similar conditions used in the
preparation of compound 2-6 (see Example 2). Purification of the
crude product by flash chromatography (silica gel, 85:15
hexanes/EtOAc) gave desired product as bright yellow foam: .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 16.02 (s, 1H), 7.52-7.26 (m,
10H), 6.77 (s, 1H), 5.36 (s, 2H), 4.90 (s, 2H), 3.97 (br s, 3H),
3.12 (dd, J=16.5, 4.9 Hz, 1H), 2.98-2.88 (m, 1H), 2.60-2.40 (m,
9H), 2.16-2.10 (m, 1H), 0.83 (s, 9H), 0.28 (s, 3H), 0.14 (s, 3H);
MS (ESI) m/z 806.5 (M+H), calcd for
C.sub.42H.sub.47F.sub.3N.sub.3O.sub.8Si 806.30.
[0565] Compound 264. Compound 264 was prepared by compound 11-6 and
propionaldehyde by similar procedures used in the preparation of
Compound 180.The crude product was purified by HPLC on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu.,
RP-.gamma. 100 R column [30.times.21.20 mm, 10 micron; flow rate,
20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH3CN; injection
volume: 4.0 mL (0.05 N HCl/water); gradient: 15.fwdarw.60% B over
15 min; mass-directed fraction collection]. Fractions with the
desired MW, eluting at 16.95-18.25 min, were collected and
freeze-dried to give product as a yellow solid: .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 7.36 (s, 1H), 4.12 (s, 1H), 3.36-3.25 (m,
2H), 3.20-2.90 (m, 9H), 2.39-2.21 (m, 2H), 1.78-1.58 (m, 3H), 1.04
(t, J=7.3 Hz, 3H); MS (ESI) m/z 556.4 (M+H), calcd for
C.sub.25H.sub.29F.sub.3N.sub.3O.sub.8 556.18.
[0566] Compound 265. Compound 265 was prepared from 11-6 and
isovaleraldehyde by similar procedures used in the preparation of
Compound 180. The crude product was purified by HPLC on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu.
RP-.gamma. 100 R column [30.times.21.20 mm, 10 micron; flow rate,
20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH.sub.3CN;
injection volume: 4.0 mL (0.05 N HCl/water); gradient:
15.fwdarw.60% B over 15 min; mass-directed fraction collection].
Fractions with the desired MW, eluting at 16.75-18.30 min, were
collected and freeze-dried to give product as a yellow solid:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.32 (s, 1H), 4.11 (s,
1H), 3.48-3.30 (m, 2H), 3.20-2.90 (m, 9H), 2.38-2.21 (m, 2H),
1.80-1.58 (m, 4H), 0.98 (d, J=6.4 Hz, 6H); MS (ESI) m/z 584.4
(M+H), calcd for C.sub.27H.sub.33F.sub.3N.sub.3O.sub.8 584.21.
[0567] Compound 266. Compound 266 was prepared from 11-6 and
ethanesulfonyl chloride by similar procedures used in the
preparation of Compound 153. The crude product was purified by HPLC
on a Waters Autopurification system using a Phenomenex Polymerx.TM.
10.mu. RP-.gamma. 100 R column [30.times.21.20 mm, 10 micron; flow
rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B:
CH.sub.3CN; injection volume: 4.0 mL (0.05 N HCl/water); gradient:
20.fwdarw.60% B over 15 min; mass-directed fraction collection].
Fractions with the desired MW, eluting at 11.10-12.00 min, were
collected and freeze-dried to give product as a yellow solid:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.62 (s, 1H), 4.09 (s,
1H), 3.18-2.85 (m, 11H), 2.36-2.19 (m, 2H), 1.69-1.59 (m, 1H), 1.35
(t, J=7.3 Hz, 3H); MS (ESI) m/z 606.3 (M+H), calcd for
C.sub.24H.sub.27F.sub.3N.sub.3O.sub.10S 606.13.
[0568] Compound 267. Compound 267 was prepared from compound 11-6
and isobutanesulfonyl chloride by similar procedures used in the
preparation of Compound 153. The crude product was purified by HPLC
on a Waters Autopurification system using a Phenomenex Polymerx.TM.
10.mu. RP-.gamma. 100 R column [30.times.21.20 mm, 10 micron; flow
rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B:
CH.sub.3CN; injection volume: 4.0 mL (0.05 N HCl/water); gradient:
20.fwdarw.60% B over 15 min; mass-directed fraction collection].
Fractions with the desired MW, eluting at 13.40-14.50 min, were
collected and freeze-dried to give product as a yellow solid:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.61 (s, 1H), 4.10 (s,
1H), 3.18-2.90 (m, 11H), 2.28-2.18 (m, 3H), 1.69-1.59 (m, 1H), 1.05
(d, J=6.8 Hz, 6H); MS (ESI) m/z 634.4 (M+H), calcd for
C.sub.26H.sub.31F.sub.3N.sub.3O.sub.10S 634.16.
[0569] Compound 268. Compound 268 was prepared from compound 11-6
and benzenesulfonyl chloride by similar procedures used in the
preparation of Compound 153. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.80 (d, J=8.0 Hz, 2H), 7.63 (s, 1H), 7.58-7.56 (m, 1H),
7.47 (t, J=8.0 Hz, 2H), 4.14 (s, 1H), 3.12-2.90 (m, 9H), 2.29-2.12
(m, 2H), 1.65-1.53 (m, 1H); MS (ESI) m/z 654.1 (M+H).
Example 12
Synthesis of Compounds of Formula IV
##STR00278##
[0571] In Scheme 12, "PG" represents a protecting group; "R.sup.F"
represents benzyl or BOC; "R.sup.F'" represents benzyl or hydrogen;
and R.sup.G represents optionally substituted heterocyclyl.
[0572] 1-methoxy-4-(trifluoromethyObenzene (12-2). To an acetone
solution of 4-(Trifluoromethyl)phenol (12-1, 125 g, 0.71 mol) was
added potassium carbonate (137 g, 0.99 mol, 1.4 equiv) and MeI (125
mL, 1.99 mol, 2.8 equiv). The reaction was stirred at 25.degree. C.
overnight and concentrated. The resulting mixture was diluted with
500 mL of H.sub.2O and extracted with t-butylmethyl ether. The
combined extracts were dried (Na.sub.2SO.sub.4) and concentrated to
give 110 g of crude 12-2: .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.48 (d, J=8.5 Hz, 2H), 6.89 (d, J=8.5 Hz, 2H), 3.78 (s, 3H).
[0573] 2-methoxy-5-(trifluoromethyObenzoic acid (12-3). nBuLi (276
mL, 2.5 M, 0.69 mol, 1.1 equiv) and KOtBu (77 g, 0.69 mol, 1.1
equiv) were added to a THF solution (1 L) of crude 12-2 (110 g) at
-78.degree. C. The reaction was stirred at -78.degree. C. for 4 h.
Dry CO.sub.2 was bubbled into the solution at -78.degree. C. for 1
h. The reaction was allowed to warm to 25.degree. C. over 0.5 h.
NaOH (6N, 300 mL) and H.sub.2O were added to the resulting reaction
mixture. The mixture was extracted with t-butylmethyl ether (400
mL.times.2). The aqueous layer was acidified with HCl (6N) to pH 1
and extracted with EtOAc (300 mL.times.4). The combined EtOAc
extracts were concentrated and re-dissolved in CH.sub.2Cl.sub.2.
The solution was dried (Na.sub.2SO.sub.4), filtered through a pad
of Celite and concentrated. Flash chromatography on silica gel (1:1
PE/EtOAc) yielded 82 g of compound 12-3 (53% for 2 steps): .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 8.45 (s, 1H), 7.81 (d, J=8.0 Hz,
1H), 7.16 (d, J=8.0 Hz, 1H), 4.13 (s, 3H); MS (ESI) m/z 221.0
(M+H).
[0574] 6-methoxy-2-methyl-3-(trifluoromethyl)benzoic acid (12-4).
To a THF solution of 2,2,6,6-tetramethylpiperidine (312 mL, 1.85
mol, 5 equiv) was added nBuLi (740 mL, 2.5 M, 1.85 mol, 5 equiv).
The reaction was stirred at 0.degree. C. for 45 min. To the
reaction was added a THF solution (800 mL) of crude 12-3 (82 g,
0.37 mol) at 0.degree. C. The reaction was stirred at 0.degree. C.
for 4 h. The reaction was cooled to -78.degree. C. and added
dropwise to a THF solution (200 mL) of MeI (200 mL, 3.73 mol, 10
equiv) at -78.degree. C. The reaction was allowed to warm to
25.degree. C. over 1 h. The reaction was quenched with HCl (6 N)
and extracted with EtOAc. The combined EtOAc extracts were dried
(Na.sub.2SO.sub.4) and concentrated to give the crude product. The
crude product was purified by HPLC. Yielded 45g of compound 12-4
(52%): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.64 (d, J=8.0 Hz,
1H), 6.81 (d, J=8.0 Hz, 1H), 3.90 (s, 3H), 2.48 (s, 3H); MS (ESI)
m/z 235.1 (M+H).
[0575] Phenyl 6-methoxy-2-methyl-3-(trifluoromethyl)benzoate
(12-5). Oxalyl chloride (66 mL, 0.77 mol, 4 equiv) was added to a
CH.sub.2Cl.sub.2 solution (500 mL, anhydrous) of compound 12-4 (45
g, 0.192 mol). DMF (0.5 mL) was added to the resulting mixture. The
reaction was stirred at 25.degree. C. for 1 h and concentrated. The
resulting solid was re-dissolved in 500 mL of anhydrous
CH.sub.2Cl.sub.2. Phenol (36.1 g, 0.38 mol, 2 equiv), DMAP (4.6 g,
38 mmol, 0.2 equiv), and triethylamine (105 mL, 0.77 mol, 4 equiv)
were added to the reaction mixture. The reaction was stirred at
25.degree. C. for 12 h and concentrated. EtOAc and H.sub.2O were
added to the residue. The organic layer was washed with NaOH (1 N),
H.sub.2O, and brine, dried (Na.sub.2SO.sub.4), and concentrated to
give 52.0 g of crude 12-5: .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.65-7.75 (m, 1H), 7.40-7.50 (m, 3H), 7.20-7.35 (m, 2H),
6.85-6.90 (m, 1H), 3.95 (s, 3H), 2.54 (s, 3H).
[0576] Phenyl 6-hydroxy-2-methyl-3-(trifluoromethyl)benzoate
(12-6). BBr.sub.3 (95 g, 0.38 mol, 2 equiv) was added to a
CH.sub.2Cl.sub.2 solution (500 mL) of crude 12-5 (52.0 g,) at
-78.degree. C. The reaction was stirred from -78.degree. C. to
25.degree. C. for 1.5 h, quenched with saturated NaHCO.sub.3 and
concentrated. EtOAc and H.sub.2O were added to the reaction
mixture. The aqueous layer was extracted with EtOAc. The combined
EtOAc extracts were dried (Na.sub.2SO.sub.4) and concentrated to
yield 51 g of crude 12-6.
[0577] Phenyl 6-(benzyloxy)-2-methyl-3-(trifluoromethyl)benzoate
(12-7-1). NaH (466 mg, 60% in oil, 11.6 mmol, 1.2 equiv) was added
to a DMF solution (30 mL) of crude 12-6 (3 g). The reaction was
stirred at 25.degree. C. for 0.5 h. To the resulting reaction was
added BnBr (1.6 mL, 13.6 mmol, 1.4 equiv) and stirred at 25.degree.
C. for 12 h. The resulting mixture was quenched with H.sub.2O and
extracted with EtOAc. The combined EtOAc extracts were washed with
NaOH (1N), H.sub.2O, and brine, dried (Na.sub.2SO.sub.4), and
concentrated to give the crude product. Flash chromatography on
silica gel (40:1 hexanes/EtOAc) yielded 3.0 g of compound 12-7-1
(81% 2 steps) as a light yellow solid: .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.60-7.70 (m, 1H), 7.15-50 (m, 8H), 7.03-7.10
(m, 2H), 6.85-6.95 (m, 1H), 5.19 (s, 2H), 2.52 (s, 3H).
[0578] Phenyl
6-(tert-butoxycarbonyloxy)-2-methyl-3-(trifluoromethyl)benzoate
(12-7-2). Boc.sub.2O (82.8 g, 0.38 mol, 2 equiv), DMAP (4.6 g, 38
mmol, 0.2 equiv) were added to a CH.sub.2Cl.sub.2 solution (150 mL)
of crude 12-6 (51 g). The reaction was stirred at 25.degree. C. for
3 h. The resulting mixture was quenched with H.sub.2O and extracted
with EtOAc.
[0579] The combined EtOAc extracts were washed with brine, dried
(Na.sub.2SO.sub.4), and concentrated to give the crude product.
Flash chromatography on silica gel (30:1 PE/EtOAc) yielded 40.0 g
of compound 12-7-2 (53% 2 steps) as a white solid: .sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 7.76 (d, J=8.0 Hz, 1H), 7.45-7.41 (m, 2H),
7.30-7.24 (m, 4H), 2.58 (s, 3H), 1.46 (s, 9H); MS (ESI) m/z 397.2
(M+H).
[0580]
(4aS,11aR,12aS,13S)-3,7-bis(benzyloxy)-4a-(tert-butyldimethylsilylo-
xy)-13-(dimethylamino)-5-hydroxy-10-(trifluoromethyl)-11a,12,12a,13-tetrah-
ydrotetraceno[2,3-d]isoxazole-4,6(4aH,11H)-dione (12-8-1). A THF
solution (3 mL) of 12-7-1 (100 mg, 0.26 mmol, 1.5 equiv) and enone
1-6 (84 mg, 0.17 mmol) was added TMEDA (0.23 mL, 1.56 mmol, 9.2
equiv) at -78.degree. C. LDA (2.2 mL, 10% wt suspension, 1.5 mmol,
8.8 equiv) was added to the reaction at -78.degree. C. The reaction
was stirred at -78.degree. C. for 30 min and allowed to warm to
25.degree. C. over 1 h, quenched with saturated NH.sub.4Cl, and
extracted with EtOAc. The combined EtOAc extracts were dried
(Na.sub.2SO.sub.4) and concentrated to yield the crude product.
[0581] Preparative reverse phase HPLC purification on a Waters
Autopurification system using a Sunfire Prep C18 OBD column [5
.mu.m, 19.times.50 mm; flow rate, 20 mL/min; Solvent A: H.sub.2O
with 0.1% HCO.sub.2H; Solvent B: CH.sub.3CN with 0.1% HCO.sub.2H;
injection volume: 4.0 mL (CH.sub.3CN); gradient: 80.fwdarw.100% B
over 15 min; mass-directed fraction collection]. Fractions with the
desired MW, eluting at 7.8-9.2 min, were collected and concentrated
on a RotaVap at 25.degree. C. to remove most of the acetonitrile.
The resulting mostly aqueous solution was extracted with EtOAc. The
combined EtOAc extracts were dried (Na.sub.2SO.sub.4) and
concentrated to give 35 mg of pure 12-8-1 (26%).
[0582]
(4aS,11aR,12aS,13S)-3-(benzyloxy)-4a-(tert-butyldimethylsilyloxy)-1-
3-(dimethylamino)-5,7-dihydroxy-10-(trifluoromethyl)-11a,12,12a,13-tetrahy-
drotetraceno[2,3-dfisoxazole-4,6(4aH,11H)-dione (12-8-2). A THF
solution (3 mL) of 12-7-2 (500 mg, 1.26 mmol, 3 equiv) and enone
(203 mg, 0.42 mmol) was added TMEDA (0.75 mL, 3.78 mmol, 30 equiv)
at -78.degree. C. LDA (2.52 mL, 1M, 2.52 mmol, 6 equiv) was added
to the reaction at -78.degree. C. The reaction was stirred at
-78.degree. C. for 30 min and allowed to warm to 25.degree. C. over
1 h, quenched with saturated NH.sub.4Cl, and extracted with EtOAc.
The combined EtOAc extracts were dried (Na.sub.2SO.sub.4) and
concentrated to yield the crude product. Preparative reverse phase
HPLC purification on a Waters Autopurification system using a
Sunfire Prep C18 OBD column and freeze-dried to give 247 mg of pure
12-8-2 (75%): .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 13.95 (s,
1H), 12.15 (s, 1H), 7.64 (d, J=9.2 Hz, 1H), 7.43(d, J=7.2 Hz, 2H),
7.34-7.28 (m, 3H), 6.84 (d, J=8.8 Hz, 1H), 5.30 (s, 2H), 3.82 (d,
J=10.4 Hz, 1H), 3.31-3.25 (m, 1H), 3.05-2.90 (m, 1H), 2.71-2.62 (m,
1H); 2.43 (s, 8H), 0.80 (s, 9H), 0.22 (s, 3H), 0.08 (s, 3H) MS
(ESI) m/z 785.3 (M+H).
[0583]
(4S,4aS,5aR,12aS)-4-(dimethylamino)-3,10,12,12a-tetrahydroxy-1,11-d-
ioxo-7-(trifluoromethyl)-1,4,4a,5,5a,6,11,12a-octahydrotetracene-2-carboxa-
mide (Compound 269). Aqueous HF (1 mL, 48%) was added to a
CH.sub.3CN solution (4 mL) of 12-8 (35 mg, 0.045 mmol) in a
polypropylene tube at 25.degree. C. The reaction was stirred at
25.degree. C. for 18 h. The resulting mixture was poured into an
aqueous solution of K.sub.2HPO.sub.4 (5 g, dissolved in 30 mL
water). The mixture was extracted with EtOAc. The combined EtOAc
extracts were dried (Na.sub.2SO.sub.4) and concentrated to yield 35
mg of crude intermediate.
[0584] Palladium on carbon (10 mg, 10 wt %) was added to a HCl/MeOH
solution (0.5N, 2 mL) of the above crude intermediate (35 mg). The
reaction was purged with hydrogen and stirred under H.sub.2
(balloon) at 25.degree. C. for 4 h. The reaction mixture was
filtered through a small Celite plug. The filtrate was concentrated
to yield the crude product. Preparative reverse phase HPLC
purification on a Waters Autopurification system using a Phenomenex
Polymerx.TM. 10.mu. RP-1 100A column [10 .mu.m, 150.times.21.20 mm;
flow rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B:
CH.sub.3CN; injection volume: 4.0 mL (0.05 N HCl/water); gradient:
0.fwdarw.50% B over 7 min, 50.fwdarw.100% over 3 min, and 100% over
5 min; mass-directed fraction collection]. Fractions with the
desired MW, eluting at 6.4-8.2 min, were collected and freeze-dried
to yield 6 mg of Compound 269 (28% for 2 steps): .sup.1H NMR (400
MHz, CD.sub.3OD) .delta. 7.76 (d, J=8.8 Hz, 1H), 6.95 (d, J=8.8 Hz,
1H), 4.10 (s, 1H), 3.25-2.93 (m, 3H), 3.04 (s, 3H), 2.95 (s, 3H),
2.60-2.48 (m, 1), 2.24-2.17 (m, 1H), 1.70-1.58 (m, 1H); MS (ESI)
m/z 483.21 (M+H).
[0585] Alternatively, Compound 269 was prepared from 12-8-2.
Aqueous HF (10 mL, 40%) was added to a THF solution (10 mL) of
12-8-2 (247 mg, 0.43 mmol) in a polypropylene tube at 25.degree. C.
The reaction was stirred at 25.degree. C. overnight. The resulting
mixture was poured into an aqueous solution of K.sub.2HPO.sub.4 (20
g, dissolved in 100 mL water). The mixture was extracted with
EtOAc. The combined EtOAc extracts were dried (Na.sub.2SO.sub.4)
and concentrated to yield 250 mg of the crude intermediate.
[0586] Palladium on carbon (150 mg, 10 wt %) was added to a
HCl/MeOH solution (0.5N, 6 mL) of the above crude intermediate (250
mg). The reaction was purged with hydrogen and stirred under
H.sub.2 (balloon) at 25.degree. C. for 1 h. The reaction mixture
was filtered through a small Celite plug. The filtrate was
concentrated to yield the crude product. Preparative reverse phase
HPLC purification on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-1 100A column and freeze-dried to
yield 105 mg of Compound 269 (51% for 2 steps).
[0587]
(4S,4aS,5aR,12aS)-9-amino-4-(dimethylamino)-3,10,12,12a-tetrahydrox-
y-1,11-dioxo-7-(trifluoromethyl)-1,4,4a,5,5a,6,11,12a-octahydrotetracene-2-
-carboxamide (Compound 270). A mixture of HNO.sub.3 (1 .mu.L, 69%)
and H.sub.2SO.sub.4 (0.2 mL) was added to a H.sub.2SO.sub.4
solution (0.5 mL) of Compound 269 (6 mg, 0.012 mmol) at 0.degree.
C. The reaction was stirred at 0.degree. C. for 30 min. The
resulting mixture was added dropwise to vigorously stirred diethyl
ether (20 mL). The suspension was filtered through a small Celite
pad and washed several times with more diethyl ether. The Celite
pad was then eluted with MeOH until the eluent became colorless.
The yellow MeOH eluent was collected and concentrated under reduced
pressure to afford the crude intermediate.
[0588] Palladium on carbon (2 mg, 10 wt %) was added to a MeOH
solution (1 mL) of the above crude intermediate. The reaction was
purged with hydrogen and stirred under H.sub.2 (balloon) at
25.degree. C. for 2 h. The catalyst was filtered off with a small
Celite pad.
[0589] The filtrate was concentrated to yield the crude product.
Preparative reverse phase HPLC purification on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu. RP-1
100A column [10 .mu.m, 150.times.21.20 mm; flow rate, 20 mL/min;
Solvent A: 0.05 N HCl; Solvent B: CH.sub.3CN; injection volume: 4.0
mL (0.05 N HCl/water); gradient: 0.fwdarw.50% B over 7 min,
50.fwdarw.400% over 3 min, and 100% over 5 min; mass-directed
fraction collection]. Fractions with the desired MW, eluting at
10-12 min, were collected and freeze-dried to yield 3 mg of pure
Compound 270 (49% for 2 steps): .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.75 (s, 1H), 4.12 (s, 1H), 3.25-2.92 (m, 3H), 3.03 (s,
3H), 2.95 (s, 3H), 2.64-2.54 (m, 1H), 2.27-2.20 (m, 1H), 1.71-1.60
(m, 1H); MS (ESI) m/z 498.22 (M+H).
[0590] Compound 271. Anhydrous Na.sub.2CO.sub.3 (16 mg, 0.15 mmol,
6.3 equiv) was added to an anhydrous DMPU/acetonitrile (150
.mu.L/50 .mu.L) solution of Compound 270 (11 mg, 0.024 mmol).
Bromoacetyl bromide (2.5 .mu.L, 0.029 mmol, 1.2 equiv) was added to
the mixture. The reaction was stirred at 25.degree. C. for 10 min.
Pyrrolidine (19 .mu.L, 0.24 mmol, 10 equiv) was added to the
reaction mixture. The reaction was stirred at 25.degree. C. for 2
h. The reaction mixture was concentrated and acidified with HCl
(0.5 N in MeOH, 0.7 mL). The resulting mixture was added dropwise
to vigorously stirred diethyl ether (10 mL). The suspension was
filtered through a small Celite pad and washed several times with
more diethyl ether. The Celite pad was then eluted with MeOH until
the eluent became colorless. The yellow MeOH eluent was collected
and concentrated under reduced pressure to afford the crude
product. Preparative reverse phase HPLC purification on a Waters
Autopurification system using a Phenomenex Polymerx.TM. 10.mu. RP-1
100A column [10 .mu.m, 150.times.21.20 mm; flow rate, 20 mL/min;
Solvent A: 0.05 N HCl; Solvent B: CH.sub.3CN; injection volume: 2.0
mL (0.05 N HCl/water); gradient: 0.fwdarw.50% B over 30 min;
mass-directed fraction collection]. Fractions with the desired MW
were collected and freeze-dried to yield 2.0 mg of pure Compound
271: .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.70 (s, 1H), 4.33
(s, 2H), 4.11 (s, 1H), 3.84-3.75 (m, 2H), 3.30-2.90 (m, 5H), 3.03
(s, 3H), 2.95 (s, 3H), 2.60-2.50 (m, 1H), 2.26-2.00 (m, 5H),
1.71-1.58 (m, 1H); MS (ESI) m/z 609.26 (M+H).
[0591] Compounds 271-304 were prepared similarly substituting the
appropriate amine NHR.sup.3R.sup.3.
[0592] Compound 272. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.72
(s, 1H), 4.11 (s, 1H), 4.09 (s, 2H), 3.22-2.90 (m, 3H), 3.03 (s,
3H), 2.92 (s, 3H), 2.59-2.49 (m, 1H), 2.25-2.18 (m, 1H), 1.70-1.59
(m, 1H), 1.42 (s, 9H); MS (ESI) m/z 611.25 (M+H).
[0593] Compound 273. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.69
(s, 1H), 4.25 (s, 2H), 4.11 (s, 1H), 3.25-2.88 (m, 3H), 3.03 (s,
3H), 3.01 (s, 6H), 2.95 (s, 3H), 2.59-2.49 (m, 1H), 2.25-2.17 (m,
1H), 1.70-1.58 (m, 1H); MS (ESI) m/z 583.23 (M+H).
[0594] Compound 274. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.70
(s, 1H), 4.22 (s, 2H), 4.12 (s, 1H), 3.23-2.92 (m, 3H), 3.05 (s,
3H), 2.96 (s, 3H), 2.91-2.84 (m, 1H), 2.60-2.49 (m, 1H), 2.27-2.19
(m, 1H), 1.71-1.59 (m, 1H), 1.00-0.92 (m, 4H); MS (ESI) m/z 595.24
(M+H).
[0595] Compound 275. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.71
(s, 1H), 4.13 (s, 1H), 4.11 (s, 2H), 3.70-3.60 (m, 1H), 3.22-2.93
(m, 3H), 3.05 (s, 3H), 2.95 (s, 3H), 2.59-2.49 (m, 1H), 2.26-2.10
(m, 3H), 1.90-1.79 (m, 2H), 1.78-1.60 (m, 5H); MS (ESI) m/z 623.29
(M+H).
[0596] Compound 276. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.72
(s, 1H), 4.12 (s, 3H), 3.25-2.94 (m, 3H), 3.05 (s, 3H), 2.97 (s,
2H), 2.95 (s, 3H), 2.60-2.49 (m, 1H), 2.25-2.20 (m, 1H), 1.71-1.59
(m, 1H), 1.12 (s, 9H); MS (ESI) m/z 625.31 (M+H).
[0597] Compound 277. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.72
(s, 1H), 4.12 (s, 1H), 4.10 (s, 2H), 3.25-2.92 (m, 5H), 3.04 (s,
3H), 2.95 (s, 3H), 2.60-2.49 (m, 1H), 2.26-2.18 (m, 1H), 2.12-2.03
(m, 1H), 1.71-1.60 (m, 1H), 1.07 (d, J=6.7 Hz, 6H); MS (ESI) m/z
611.28 (M+H).
[0598] Compound 278. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.72
(s, 1H), 4.13 (s, 2H), 4.16 (s, 1H), 3.23-2.93 (m, 5H), 3.04 (s,
3H), 2.96 (s, 3H), 2.60-2.50 (m, 1H), 2.25-2.19 (m, 1H), 1.71-1.60
(m, 1H), 1.20-1.09 (m, 1H), 0.78-0.73 (m, 1H), 0.48-0.41 (m, 1H);
MS (ESI) m/z 609.24 (M+H).
[0599] Compound 279. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.65
(s, 1H), 4.41-4.34 (m, 2H), 4.35 (s, 2H), 4.24-4.17 (m, 2H), 4.11
(s, 1H), 3.22-2.92 (m, 3H), 3.04 (s, 3H), 2.95 (s, 3H), 2.70-2.60
(m, 1H), 2.59-2.42 (m, 2H), 2.25-2.18 (m, 1H), 1.70-1.59 (m, 1H);
MS (ESI) m/z 595.21 (M+H).
[0600] Compound 280. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.71
(s, 1H), 4.11 (s, 1H), 4.09 (s, 2H), 3.24-2.94 (m, 3H), 3.05 (s,
3H), 2.96 (s, 3H), 2.80 (s, 3H), 2.60-2.50 (m, 1H), 2.26-2.19 (m,
1H), 1.71-1.60 (m, 1H); MS (ESI) m/z 569.20 (M+H).
[0601] Compound 281. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.68
(s, 1H), 4.10 (s, 3H), 3.20-2.93 (m, 10H), 2.56-2.47 (m, 1H),
2.25-2.10 (m, 3H), 1.92-1.85 (m, 2H), 1.73-1.59 (m, 2H), 1.45-1.33
(m, 4H), 1.30-1.16 (m, 1H); MS (ESI) m/z 637.1 (M+H).
[0602] Compound 282. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.67
(s, 1H), 4.30 (d, J=16.4 Hz, 1H), 4.19 (d, J=15.6 Hz, 1H), 4.10 (s,
1H), 3.21-2.93 (m, 14H), 2.56-2.48 (m, 1H), 2.22-2.19 (m, 1H),
1.84-1.75 (m, 2H), 1.67-1.57 (m, 1H), 1.02 (t, J=7.2 Hz, 3H); MS
(ESI) m/z 611.1 (M+H).
[0603] Compound 283. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.66
(d, J=8.4 Hz, 1H), 4.27 (d, J=18.8 Hz, 2H), 4.10 (s, 1H), 3.98-3.94
(m, 1H), 3.47-3.43 (m, 1H), 3.18-2.85 (m, 12H), 2.76-2.71 (m, 2H),
2.54-2.47 (m, 1H), 2.23-2.16 (m, 1H), 1.85-1.53 (m, 6H); MS (ESI)
m/z 649.1 (M+H).
[0604] Compound 284. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.65
(s, 1H), 4.07 (s, 2H), 4.03 (s, 1H), 3.97-3.90 (m, 2H), 3.12-2.86
(m, 9H), 2.49-2.42 (m, 1H), 2.15-2.11 (m, 1H), 1.57-1.50 (m, 1H),;
MS (ESI) m/z 637.1 (M+H).
[0605] Compound 285. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.72
(s, 1H), 5.50 (d, J=52.0 Hz, 1H), 4.44 (s, 2H), 4.14 (s, 1H),
4.04-3.95 (m, 2H), 3.54-3.47 (m, 2H), 3.23-2.97 (m, 9H), 2.60-2.23
(m, 4H), 1.71-1.62 (m, 1H); MS (ESI) m/z 627.0 (M+H).
[0606] Compound 286. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.70
(s, 1H), 4.11 (s, 1H), 4.09 (s, 2H), 3.51-3.44 (m, 1H), 3.19-2.94
(m, 9H), 2.56-2.49 (m, 1H), 2.24-2.19 (m, 1H), 1.68-1.58 (m, 1H),
1.38 (t, J=6.4 Hz, 6H); MS (ESI) m/z 597.2 (M+H).
[0607] Compound 287. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.69
(s, 1H), 4.10 (s, 1H), 4.07 (s, 2H), 3.21-2.94 (m, 11H), 2.56-2.50
(m, 1H), 2.25-2.19 (m, 1H), 1.85-1.60 (m, 7H), 1.37-1.22 (m, 3H),
1.12-1.03 (m, 2H); MS (ESI) m/z 651.3 (M+H).
[0608] Compound 288. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.71
(s, 1H), 4.32 (s, 2H), 4.14 (s, 1H), 4.10-3.87 (m, 4H), 3.65-3.58
(m, 2H), 3.43-3.36 (m, 2H), 3.23-2.97 (m, 9H), 2.60-2.52 (m, 1H),
2.27-2.23 (m, 1H), 1.70-1.64 (m, 1H); MS (ESI) m/z 625.3 (M+H).
[0609] Compound 289. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.72
(s, 1H), 4.14 (s, 1H), 4.13 (s, 2H), 3.19-2.90 (m, 11H), 2.56-2.45
(m, 1H), 2.23-2.15 (m, 2H), 1.92-1.85 (m, 2H), 1.70-1.65 (m, 5H),
1.31-1.22 (m, 2H); MS (ESI) m/z 637.2 (M+H).
[0610] Compound 290. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.67
(s, 1H), 5.45 (d, J=52.0 Hz, 1H), 4.40 (s, 2H), 4.09 (s, 1H),
4.00-3.90 (m, 1H), 3.45-3.37 (m, 1H), 3.20-2.92 (m, 11H), 2.53-2.33
(m, 3H), 2.23-2.15 (m, 1H), 1.65-1.55 (m, 1H); MS (ESI) m/z 627.2
(M+H).
[0611] Compound 291. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.58
(s, 1H), 5.37 (d, J=56.8 Hz, 1H), 4.36 (m, 4H), 4.02 (s, 1H),
3.17-2.73 (m, 11H), 2.48-2.41 (m, 1H), 2.15-2.11 (m, 1H), 1.60-1.50
(m, 1H); MS (ESI) m/z 613.1 (M+H).
[0612] Compound 292. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.68
(s, 1H), 4.15 (s, 1H), 4.11 (s, 2H), 3.21-2.98 (m, 11H), 2.56-2.48
(m, 1H), 2.26-2.19 (m, 1H), 1.82-1.73 (m, 2H), 1.68-1.49 (m, 1H),
1.02 (t, J=7.2 Hz, 3H); MS (ESI) m/z 597.2 (M+H).
[0613] Compound 293. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.61
(s, 1H), 4.03 (s, 1H), 3.90 (s, 2H), 3.80-3.72 (m, 1H), 3.15-2.87
(m, 9H), 2.48-2.40 (m, 1H), 2.37-2.11 (m, 5H), 1.90-1.81 (m, 2H),
1.62-1.52 (m, 1H); MS (ESI) m/z 609.2 (M+H).
[0614] Compound 294. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.65
(s, 1H), 4.10 (s, 3H), 3.57 (t, J=5.2 Hz, 2H), 3.48 (s, 3H),
3.30-3.25 (m, 2H), 3.18-2.92 (m, 9H), 2.55-2.45 (m, 1H), 2.23-2.15
(m, 1H), 1.65-1.55 (m, 1H); MS (ESI) m/z 613.2 (M+H).
[0615] Compound 295. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.16
(s, 1H), 4.33 (d, J=15.6 Hz, 1H), 4.21 (d, J=16.4 Hz, 1H), 4.11 (s,
1H), 3.24-2.93 (m, 14H), 2.58-2.49 (m, 1H), 2.28-2.20 (m, 1H),
1.70-1.58 (m, 1H), 1.40 (t, J=7.4 Hz, 3H); MS (ESI) m/z 597.0
(M+H).
[0616] Compound 296. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.57
(s, 1H), 4.19 (s, 2H), 4.05 (s, 1H), 3.35-3.23 (m, 4H), 3.13-2.82
(m, 9H), 2.48-2.39 (s, 1H), 2.28-2.11 (s, 1H), 1.60-1.51 (m, 1H),
1.29 (t, J=7.2 Hz, 6H); MS (ESI) m/z 611.1 (M+H).
[0617] Compound 297. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.67
(s, 1H), 4.21 (s, 2H), 4.12 (s, 1H), 3.64-3.61 (m, 2H), 3.19-2.91
(m, 11H), 2.56-2.46 (m, 1H), 2.25-2.17 (m, 1H), 1.97-1.79 (m, 5H),
1.69-1.50 (m, 2H); MS (ESI) m/z 623.2 (M+H).
[0618] Compound 298. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.72
(s, 1H), 7.42 (s, 4H), 5.10-5.00 (m, 2H), 4.73-4.62 (m, 2H), 4.57
(s, 2H), 4.13 (s, 1H), 3.21-2.90 (m, 9H), 2.56-1.95 (m, 1H),
2.25-2.20 (m, 1H), 1.68-1.58 (m, 1H); MS (ESI) m/z 657.1 (M+H).
[0619] Compound 299. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.68
(s, 1H), 4.12 (s, 1H), 4.09 (s, 2H), 3.22-2.91 (m, 11H), 2.56-2.47
(m, 1H), 2.26-2.17 (m, 1H), 1.69-1.58 (m, 1H), 1.37 (t, J=7.2 Hz,
3H); MS (ESI) m/z 583.1 (M+H).
[0620] Compound 300. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.70
(s, 1H), 6.36 (tt, J=53.5, 3.2 Hz, 1H), 4.23 (s, 2H), 4.13 (s, 1H),
3.69 (td, J=15.3, 2.8 Hz, 2H), 3.23-2.91 (m, 9H), 2.58-2.50 (m,
1H), 2.25-2.19 (m, 1H), 1.70-1.60 (m, 1H); MS (ESI) m/z 619.0
(M+H).
[0621] Compound 301. iH NMR (400 MHz, CD.sub.3OD) .delta. 8.79 (s,
1H), 4.32 (s, 1H), 4.32 (s, 2H), 3.37-3.15 (m, 11H), 2.97-2.89 (m,
1H), 2.78-2.68 (m, 1H), 2.47-2.35 (m, 3H), 2.27-2.18 (m,1H),
2.16-2.03 (m, 3H), 1.90-1.79 (m,1H); MS (ESI) m/z 623.2 (M+H).
[0622] Compound 302. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.86
(s, 1H), 4.90-4.80 (m, 1H), 4.62-4.50 (m, 5H), 4.32 (s, 1H),
4.30-4.20 (m, 1H), 3.56(s, 3H), 3.40-3.12 (m, 9H), 2.80-2.68 (m,
1H), 2.48-2.39 (m, 1H), 1.90-1.79 (m,1H); MS (ESI) m/z 625.2
(M+H).
[0623] Compound 303. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.73
(s, 1H), 4.75 (tt, J=52.0, 4.8 Hz, 2H), 4.21 (s, 2H), 4.15 (s, 1H),
3.55 (tt, J=26.8, 4.4 Hz, 2H), 3.23-2.87 (m, 9H), 2.60-2.50 (m,
1H), 2.30-2.20 (m, 1H), 1.68-1.64 (m, 1H); MS (ESI) m/z 601.1
(M+H).
[0624] Compound 304. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.79
(s, 1H), 7.49-7.45 (m, 2H), 7.30-7.28 (m, 3H), 4.71 (s, 2H), 4.11
(s, 1H), 3.18-2.95 (m, 9H), 2.60-2.49 (m, 1H), 2.27-2.18 (m, 1H),
1.70-1.61 (m, 1H); MS (ESI) m/z 631.0 (M+H).
[0625] Compounds 307-311 were prepared similarly to 271
substituting bromoacetyl bromide and an amine with an acid chloride
(appropriately protected if needed) or a carboxylic acid
(appropriately protected if needed) and a coupling reagent such as
HATU.
[0626] Compound 307. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.71
(s, 1H), 5.32-5.28 (m, 1H), 4.21-4.17 (m, 1H), 4.14 (s, 1H),
4.07-4.00(m, 1H), 3.25-2.92 (m, 10H), 2.70-2.61 (m, 1H), 2.59-2.49
(m, 1H), 2.28-2.20 (m, 1H), 1.70-1.60 (m,1H); MS (ESI) m/z 581.1
(M+H).
[0627] Compound 308. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.54
(s, 1H), 4.07 (s, 2H), 3.46-3.38 (m, 1H), 3.16-2.90 (m, 10H),
2.53-2.43 (m, 1H), 2.33-2.15 (m, 2H), 1.96-1.86 (m, 2H), 1.77-1.55
(m, 4H); MS (ESI) m/z 609.1 (M+H).
[0628] Compound 309. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.59
(s, 1H), 5.17-5.12 (m, 1H), 4.11-3.98 (m, 3H), 3.13-2.85 (m, 12H),
2.59-2.42 (m, 3H), 2.18-2.11 (m, 1H), 1.61-1.50 (m,1H); MS (ESI)
m/z 595.1 (M+H).
[0629] Compound 310. .sup.iH NMR (400 MHz, CD.sub.3OD) .delta. 8.59
(s, 1H), 4.62-4.57 (m, 1H), 4.12 (s, 1H), 3.51-3.36 (m, 2H),
3.25-2.93 (m, 3H), 3.05 (s, 3H), 2.96 (s, 3H), 2.62-2.49 (m, 2H),
2.25-2.08 (m, 4H), 1.71-1.58 (m, 1H); MS (ESI) m/z 595.25
(M+H).
[0630] Compound 311. .sup.iH NMR (400 MHz, CD.sub.3OD) .delta. 8.61
(s, 1H), 4.42-4.37 (m, 1H), 4.13 (s, 1H), 3.82-3.75 (m, 2H),
3.23-2.92 (m, 3H), 3.04 (s, 3H), 3.01 (s, 3H), 2.95 (s, 3H),
2.78-2.62 (m, 1H), 2.62-2.51 (m, 1H), 2.32-2.04 (m, 4H), 1.71-1.60
(m, 1H); MS (ESI) m/z 609.26 (M+H).
[0631] Compound 305. To a solution of Compound 270 (15 mg, 0.024
mmol, 1 equiv) in THF(1 ml) was added sodium carbonate (20 mg, 0.19
mmol, 8 equiv), 3-fluorobenzenesulfonyl chloride (0.036 mmol, 1.5
equiv). The reaction was stirred at rt for 6 h, LC/MS analysis
indicated complete consumption of the starting aniline 12. The
reaction mixture was then added dropwise into 4 N HCl/MeOH (1 ml)
at 0.degree. C. with rapid stirring. The reaction mixture was
concentrated and the residue purified by HPLC. The desired product
Compound 305 was obtained as a yellow solid after freeze-drying:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.92 (s, 1H), 7.61-7.50
(m, 3H), 7.38-7.32 (m, 1H), 4.10 (s, 1H), 3.19-2.90 (m, 9H),
2.55-2.45 (m, 1H), 2.24-2.46 (m, 1H), 1.68-1.55 (m,1H); MS (ESI)
m/z 656.0 (M+H).
[0632] Compound 306. Compound 306 was prepared similarly to
Compound 305 using 1-methyl-1H-pyrazole-3-sulfonyl chloride:
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.82 (s, 1H), 8.06 (s,
1H), 7.93 (s, 1H), 4.15 (s, 1H), 3.91 (s, 3H), 3.96-2.95 (m, 9H),
2.59-2.49 (m, 1H), 2.28-2.20 (m, 1H), 1.70-1.60 (m,1H); MS (ESI)
m/z 642.1 (M+H).
Example 13
Synthesis of Compounds of Formula V
##STR00279##
[0634] In Scheme 13, "R.sup.F" represents benzyl or BOC; and
"R.sup.F" represents benzyl or hydrogen.
[0635] Ethyl 3-bromo-6-methoxy-2-methylbenzoate (13-2). To a
methylene chloride solution (100 mL) of Ethyl
2-methoxy-6-methylbenzoate (13-1, 5 g, 25.8 mmol, TCI M1120) was
added bromine (1.65 mL, 32.2 mmol, 1.25 equiv) at 0.degree. C. The
reaction was stirred at 0.degree. C. to 25.degree. C. for 12 h and
quenched with sodium thiosulfate solution (5%, 20 mL) and stirred
for 20 min. The resulting mixture was extracted with methylene
chloride. The combined methylene chloride extracts were dried
(Na.sub.2SO.sub.4) and concentrated to give 7.2 g of 13-2: .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.47 (d, J=9.2 Hz, 1H), 6.63 (d,
J=9.2 Hz, 1H), 4.38 (q, J=7.3 Hz, 2H), 3.78 (s, 3H), 2.30 (s, 3H),
1.35 (t, J=7.3 Hz, 3H).
[0636] Ethyl 3-bromo-6-hydroxy-2-methylbenzoate (13-3). BBr3 (31
mL, 1.0 M, 30.96 mmol, 1.2 equiv) was added to a methylene chloride
solution (100 mL) of crude 13-2 (7.2 g) at -78.degree. C. The
reaction was allowed to warm to 25.degree. C. over 2 h. The
reaction was stirred from -78.degree. C. to 25.degree. C. for 2 h,
quenched with saturated NaHCO3 and concentrated. EtOAc and H.sub.2O
were added to the reaction mixture. The aqueous layer was extracted
with EtOAc. The combined EtOAc extracts were dried
(Na.sub.2SO.sub.4) and concentrated to yield 7.1 g of 13-3: .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 10.87 (br s, 1H), 7.55 (d, J=9.2
Hz, 1H), 4.43 (q, J=7.3 Hz, 2H), 2.63 (s, 3H), 1.42 (t, J=7.3 Hz,
3H).
[0637] Ethyl 6-(benzyloxy)-3-bromo-2-methylbenzoate (13-4). To a
DMF solution (50 mL) of crude 13-3 (7.1 g) was added NaH (1.24 g,
60% in oil, 30.96 mmol, 1.2 equiv) at 0.degree. C. The reaction was
stirred at 0.degree. C. for 1 h. To the reaction mixture was added
BnBr (4.3 mL, 36.1 mmol, 1.4 equiv) at 0.degree. C. The reaction
was stirred at 25.degree. C. for 12 h. The reaction was quenched
with NH.sub.4Cl and extracted with EtOAc. The combined EtOAc
extracts were washed with H.sub.2O, brine, dried (Na.sub.2SO.sub.4)
and concentrated. Flash chromatography on silica gel (30:1
hexanes/EtOAc) yielded 8.0 g of compound 13-4 (89% for 3 steps):
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.44 (d, J=9.2 Hz, 1H),
7.25-7.40 (m, 5H), 6.66 (d, J=9.2 Hz, 1H), 5.06 (s, 2H), 4.36 (q,
J=7.3 Hz, 2H), 2.32 (s, 3H), 1.32 (t, J=7.3 Hz, 3H).
[0638] Ethyl 6-(benzyloxy)-3-cyano-2-methylbenzoate (13-5). CuCN
(770 mg, 8.6 mmol, 3 equiv) was added to a DMF solution (15 mL,
anhydrous) of 13-4 (1.0 g, 2.87 mmol) in a sealed tube. The
reaction was stirred at 200.degree. C. for 18 min and cooled. The
resulting mixture was quenched with H.sub.2O and extracted with
EtOAc. The combined EtOAc extracts were washed with H.sub.2O,
brine, dried (Na.sub.2SO.sub.4) and concentrated. Flash
chromatography on silica gel (10:1 hexanes/EtOAc) yielded 670 mg of
compound 13-5 (80%):.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.55
(d, J=8.5 Hz, 1H), 7.26-7.40 (m, 5H), 6.83 (d, J=8.5 Hz, 1H), 5.14
(s, 2H), 4.37 (q, J=7.0 Hz, 2H), 2.46 (s, 3H), 1.30 (t, J=7.0 Hz,
3H).
[0639] 6-(benzyloxy)-3-cyano-2-methylbenzoic acid (13-6). NaOH
solution (5 mL, 3 N) was added to a MeOH/THF solution (5 mL/5 mL)
of 13-5 (518 mg, 1.75 mmol) at 55.degree. C. The reaction was
stirred from 55.degree. C. for 15 h and concentrated. The resulting
mixture was acidified with HCl (6 N) to pH 1 and extracted with
EtOAc. The combined EtOAc extracts were dried (Na.sub.2SO.sub.4)
and concentrated to yield 530 mg of crude 13-6.
[0640] 6-(benzyloxy)-3-cyano-2-methylbenzoic acid (13-7-1). Oxalyl
chloride (0.75 mL, 8.75 mmol, 5 equiv) was added to a
CH.sub.2Cl.sub.2 solution (5 mL, anhydrous) of crude 13-6 (530 mg).
DMF (0.1 mL) was added to the resulting mixture. The reaction was
stirred at 25.degree. C. for 1 h and concentrated. The resulting
solid was re-dissolved in 5 mL of anhydrous CH.sub.2Cl.sub.2.
Phenol (330 mg, 3.5 mmol, 2 equiv), DMAP (213 mg, 1.75 mmol, 1
equiv), and triethylamine (1.2 mL, 8.75 mmol, 5 equiv) were added
to the reaction mixture. The reaction was stirred at 25.degree. C.
for 12 h and concentrated. EtOAc and H.sub.2O were added to the
residue. The organic layer was washed with NaOH (1 N), H.sub.2O,
and brine, dried (Na.sub.2SO.sub.4), and concentrated. Flash
chromatography on silica gel (10:1 hexanes/EtOAc) yielded 400 mg of
compound 13-7-1 (67% for 2 steps): .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.65 (d, J=9.2 Hz, 1H), 7.25-7.45 (m, 8H), 7.06
(d, J=10.2 Hz, 2H), 6.93 (d, J=9.2 Hz, 1H), 5.20 (s, 2H), 2.61 (s,
3H).
[0641] Phenyl 6-(tert-butoxycarbonyloxy)-3-cyano-2-methylbenzoate
(13-7-2). To a solution of compound 13-7-1 (5 g, 0.014 mol) in 1, 4
dioxane/MeOH (25 mL/25 mL) was added Pd/C (1.1 g, 22%), the mixture
was purged by bubbling hydrogen through for 5 min and rapidly
stirred under 1 atm hydrogen atmosphere at rt for 2 hrs. The
catalyst was filtered off with a small Celite pad and washed with
more methanol (2 mL.times.3). The filtrate was concentrated to
yield the crude intermediate. Di-tert-butyl dicarbonate (3.3 g,
0.015 mmol, 1.05 equiv), DMAP (20 mg, cat.) were added to a
solution of the above intermediate in DCM (100 mL). The resulting
mixture was stirred for 1 h at rt and concentrated. The residue was
purified by flash-column chromatography to afford the desired
product 13-7-2 as a white solid (3.2 g, 62%, two steps): .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.77 (d, J=8.8 Hz, 1H), 7.47 (m,
2H), 7.33 (m, 2H), 7.30 (m, 1H), 7.25 (d, J=7.6 Hz, 1H), 2.71 (s,
3H), 1.47 (s, 9H); MS (ESI) m/z 354.37 (M+H).
[0642]
(4aS,11aR,12aS,13S)-3,7-bis(benzyloxy)-4a-(tert-butyldimethylsilylo-
xy)-13-(dimethylamino)-5-hydroxy-4,6-dioxo-4,4a,6,11,11a,12,12a,13-octahyd-
rotetraceno[2,3-d]isoxazole-10-carbonitrile (13-8-1). A THF
solution (5 mL) of 13-7-1 (400 mg, 1.17 mmol, 1.5 equiv) and enone
1-6 (378 mg, 0.78 mmol) was added TMEDA (1.1 mL, 7.02 mmol, 6.0
equiv) at -78.degree. C. LHMDS (4.7 mL, 1 M, 4.66 mmol, 4.0 equiv)
was added to the reaction at -78.degree. C. The reaction was
stirred at -78.degree. C. for 30 min and allowed to warm to
25.degree. C. over 1 h, quenched with saturated NH.sub.4Cl, and
extracted with EtOAc. The combined EtOAc extracts were dried
(Na.sub.2SO.sub.4) and concentrated to yield the crude product.
[0643] Preparative reverse phase HPLC purification on a Waters
Autopurification system using a Sunfire Prep C18 OBD column [5
.mu.m, 19.times.50 mm; flow rate, 20 mL/min; Solvent A: H.sub.2O
with 0.1% HCO.sub.2H; Solvent B: CH.sub.3CN with 0.1% HCO.sub.2H;
injection volume: 4.0 mL (CH3CN); gradient: 80.fwdarw.100% B over
15 min; mass-directed fraction collection]. Fractions with the
desired MW, eluting at 5.9-7.0 min, were collected and concentrated
on a RotaVap at 25.degree. C. to remove most of the acetonitrile.
The resulting mostly aqueous solution was extracted with EtOAc. The
combined EtOAc extracts were dried (Na.sub.2SO.sub.4) and
concentrated to give 153 mg of pure 13-8-1 (27%).
[0644]
(4aS,11aR,12aS,13S)-3-(benzyloxy)-4a-(tert-butyldimethylsilyloxy)-1-
0-cyano-13-(dimethylamino)-5-hydroxy-4,6-dioxo-4,4a,6,11,11a,12,12a,13-oct-
ahydrotetraceno[2,3-dfisoxazol-7-yltert-butyl carbonate (13-8-2).
To a solution of compound 13-7-2 (402 mg, 1.139 mmol, 2 equiv) and
enone 1-6 (282 mg, 0.58 mmol, 1 equiv) was added TMEDA (0.55 mL,
3.5 mmol, 6 equiv) at -78.degree. C. LHMDS (3.3 mL, 1 M, 3.3 mmol,
5.6 equiv) was added to the reaction mixture at -78.degree. C. The
reaction was stirred at -78.degree. C. for 30 min and allowed to
warm to 25.degree. C. over 1 h, quenched with saturated NH.sub.4Cl,
and extracted with EtOAc. The combined extracts were dried with
Na.sub.2SO.sub.4 and concentrated to yield the crude product, which
was purified by flash-column chromatography to afford the product
13-8-2 as a yellow solid (0.14 g, 32%): .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 15.41 (s, 1H), 7.85 (d, J=4.0 Hz, 1H),
7.63-7.32 (m, 5H), 7.26-7.20 (m, 1H), 5.44 (s, 2H), 4.01 (d, J=10.8
Hz,1H), 3.48 (dd, J=16.0 Hz, 4.4 Hz, 1H), 3.21-3.18 (m, 1H),
2.94-2.81 (m, 1H), 2.69-2.63 (m, 1H), 2.71-2.55 (m, 7H), 2.26 (d,
J=14.4 Hz, 1H),1.62 (s, 9H), 0.93 (s, 9H), 0.33 (s, 3H), 0.19 (s,
3H); MS (ESI) m/z 742.37 (M+H).
[0645] Compound 312. Aqueous HF (1 mL, 48%) was added to a
CH.sub.3CN solution (4 mL) of 13-8-1 (35 mg, 0.045 mmol) in a
polypropylene tube at 25.degree. C. The reaction was stirred at
25.degree. C. for 18 h. The resulting mixture was poured into an
aqueous solution of K.sub.2HPO.sub.4 (5 g, dissolved in 30 mL
water). The mixture was extracted with EtOAc. The combined EtOAc
extracts were dried (Na.sub.2SO.sub.4) and concentrated to yield
120 mg of the crude intermediate.
[0646] Palladium on carbon (50 mg, 10 wt %) was added to a HCl/MeOH
solution (0.5N, 2 mL) of the above intermediate (120 mg). The
reaction was purged with hydrogen and stirred under H.sub.2
(balloon) at 25.degree. C. for 3 h. The reaction mixture was
filtered through a small Celite plug. The filtrate was
concentrated. Preparative reverse phase HPLC purification on a
Waters Autopurification system using a Phenomenex Polymerx.TM.
10.mu. RP-1 100A column [10 .mu.m, 150.times.21.20 mm; flow rate,
20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B: CH.sub.3CN;
injection volume: 4.0 mL (0.05 N HCl/water); gradient: 0.fwdarw.70%
B over 7 min, 70.fwdarw.100% over 3 min, and 100% over 5 min;
mass-directed fraction collection]. Fractions with the desired MW
were collected and freeze-dried to yield 28 mg of Compound 312 (30%
for 2 steps): .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.76 (d,
J=9.0 Hz, 1H), 6.97 (d, J=9.0 Hz, 1H), 4.10 (s, 1H), 3.26-2.91 (m,
3), 3.04 (s, 3H), 2.96 (s, 3H), 2.71-2.62 (m, 1H), 2.27-2.20 (m,
1H), 1.72-1.62 (m, 1H); MS (ESI) m/z 410.22 (M+H).
[0647] Alternatively, Compound 312 was prepared from 13-8-2.
Compound 13-8-2 (140 mg, 0.19 mmol) was dissolved in THF (5 mL),
and HF (5 mL, 40% in water) was added. The resultant yellow
solution was stirred at rt overnight. The reaction solution was
then slowly added into a solution of K.sub.2HPO.sub.4 (3.1 g) in
water (20 mL) with rapid stirring. The mixture was extracted with
DCM (10 mL.times.3). The combine extracts were dried over sodium
sulfate and concentrated in vacuo to yield the crude
intermediate.
[0648] To a HCl/MeOH solution (0.5 N, 5 mL) of the above crude
intermediate was added Pd/C (30 mg, 20%). The mixture was purged by
bubbling hydrogen through for 5 min and rapidly stirred under 1 atm
hydrogen atmosphere at rt for 1 hr. The catalyst was filtered off
with a small Celite pad and washed with more methanol (5
mL.times.3). The filtrate was concentrated in vacuo. Preparative
HPLC purification on a Polymerx.TM. column yielded the desired
product Compound 312 as a yellow solid (56 mg, 68% two steps).
[0649] Compound 313. A mixture of HNO.sub.3 (27 .mu.L, 69%) and
H.sub.2SO.sub.4 (0.4 mL) was added to a H.sub.2SO.sub.4 solution (1
mL) of Compound 312 (150 mg, 0.34 mmol) at 0.degree. C. The
reaction was stirred at 0.degree. C. for 30 min. The resulting
mixture was added dropwise to vigorously stirred diethyl ether (20
mL). The suspension was filtered through a small Celite pad and
washed several times with more diethyl ether. The Celite pad was
then eluted with MeOH until the eluent became colorless. The yellow
MeOH eluent was collected and concentrated under reduced
pressure.
[0650] The resulting mixture was re-dissolved in MeOH (15 mL). The
Palladium on carbon (20 mg, 10 wt %) was added to the reaction. The
reaction mixture was stirred under H.sub.2 at 25.degree. C. for 30
min. The reaction mixture was filtered through a small Celite plug.
The filtrate was concentrated. Preparative reverse phase HPLC
purification on a Waters Autopurification system using a Phenomenex
Polymerx.TM. 10.mu. RP-1 100A column [10 .mu.m, 150.times.21.20 mm;
flow rate, 20 mL/min; Solvent A: 0.05 N HCl/water; Solvent B:
CH.sub.3CN; injection volume: 4.0 mL (0.05 N HCl/water); gradient:
0.fwdarw.50% B over 7 min, 50.fwdarw.100% over 3 min, and 100% over
5 min; mass-directed fraction collection]. Fractions with the
desired MW were collected and freeze-dried to yield 62 mg of
Compound 313 (40% for 2 steps): .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 7.58 (s, 1H), 4.10 (s, 1H), 3.25-2.93 (m, 3H), 3.03 (s,
3H), 2.93 (s, 3H), 2.75-2.60 (m, 1H), 2.28-2.20 (m, 1H), 1.71-1.60
(m, 1H); MS (ESI) m/z 455.23 (M+H).
[0651] Compound 314. Anhydrous Na.sub.2CO.sub.3 (16 mg, 0.15 mmol,
6.3 equiv) was added to an anhydrous DMPU/acetonitrile (150
.mu.L/50 .mu.L) solution of Compound 313 (11 mg, 0.024 mmol).
Bromoacetyl bromide (2.5 .mu.L, 0.029 mmol, 1.2 equiv) was added to
the mixture. The reaction was stirred at 25.degree. C. for 10 min.
Pyrrolidine (19 .mu.L, 0.24 mmol, 10 equiv) was added to the
reaction mixture. The reaction was stirred at 25.degree. C. for 2
h. The reaction mixture was concentrated and acidified with HCl
(0.5 N in MeOH, 0.7 mL). The resulting mixture was added dropwise
to vigorously stirred diethyl ether (10 mL). The suspension was
filtered through a small Celite pad and washed several times with
more diethyl ether. The Celite pad was then eluted with MeOH until
the eluent became colorless. The yellow MeOH eluent was collected
and concentrated under reduced pressure. Preparative reverse phase
HPLC purification on a Waters Autopurification system using a
Phenomenex Polymerx.TM. 10.mu. RP-1 100A column [10 .mu.m,
150.times.21.20 mm; flow rate, 20 mL/min; Solvent A: 0.05 N HCl;
Solvent B: CH.sub.3CN; injection volume: 2.0 mL (0.05 N HCl/water);
gradient: 0.fwdarw.50% B over 30 min; mass-directed fraction
collection]. Fractions with the desired MW were collected and
freeze-dried to yield 4.3 mg of pure 13-11-1: .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 8.62 (s, 1H), 4.34 (s, 1H), 4.12 (s, 1H),
3.3.84-3.76 (m, 2H), 3.30-2.92 (m, 5H), 3.04 (s, 3H), 2.95 (s, 3H),
2.70-2.58 (m, 1H), 2.30-2.01 (m, 5H), 1.72-1.61 (m, 1H); MS (ESI)
m/z 566.27 (M+H).
[0652] Compounds 315-333 were prepared similarly employing the
appropriate amine (NHR.sup.2R.sup.3).
[0653] Compound 315. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.64
(s, 1H), 4.12 (s,1H), 4.09 (s, 2H), 3.30-2.92 (m, 3H), 3.05 (s,
3H), 2.95 (s, 3H), 2.71-2.58 (m, 1H), 2.30-2.22 (m, 1H), 1.73-1.62
(m, 1H), 1.42 (s, 9H); MS (ESI) m/z 568.19 (M+H).
[0654] Compound 316. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.63
(s, 1H), 4.26 (s, 1H), 4.13 (s, 1H), 3.30-2.94 (m, 3H), 3.05 (s,
3H), 3.02 (s, 6H), 2.95 (s, 3H), 2.71-2.60 (m, 1H), 2.30-2.23 (m,
1H), 1.73-1.62 (m, 1H); MS (ESI) m/z 540.24 (M+H).
[0655] Compound 317. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.65
(s, 1H), 4.12 (s, 1H), 4.10 (s, 2H), 3.20-2.92 (m, 5H), 3.04 (s,
3H), 2.95 (s, 3H), 2.71-2.62 (m, 1H), 2.30-2.22 (m, 1H), 2.13-2.24
(m, 1H), 1.73-1.63 (m, 1H), 1.08 (d, J=6.7 Hz, 6H); MS (ESI) m/z
568.21 (M+H).
[0656] Compound 318. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.53
(s, 1H), 4.11 (s, 2H), 4.04 (s, 1H), 3.20-2.75 (m, 10H), 2.58-2.52
(m, 1H), 2.19-2.15 (m, 1H),1.62-1.56 (m, 1H), 0.86-0.80 (m, 4H); MS
(ESI) m/z 552.1 (M+H).
[0657] Compound 319. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.53
(s, 1H), 4.05 (s, 1H), 3.90 (s, 2H), 3.20-2.85 (m, 11H), 2.62-2.48
(m, 1H), 2.19-2.15 (m, 1H), 1.58-1.53 (m, 1H), 1.27 (t, J=7.2 Hz,
3H); MS (ESI) m/z 540.0 (M+H).
[0658] Compound 320. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.64
(s, 1H), 4.15 (s, 1H), 4.11 (s, 2H), 3.30-2.96 (m, 11H), 2.72-2.60
(m, 1H), 2.31-2.28 (m, 1H), 1.88-1.62 (m, 3H), 1.06 (t, J=7.2 Hz,
3H); MS (ESI) m/z 553.9 (M+H).
[0659] Compound 321. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.56
(s, 1H), 4.04 (s, 1H), 4.00 (s, 2H), 3.15-2.85 (m, 11H), 2.62-2.55
(m, 1H), 2.21-2.16 (m, 1H), 1.80-1.50 (m, 8H), 1.30-1.10 (m, 4H);
MS (ESI) m/z 608.0 (M+H).
[0660] Compound 322. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.54
(s, 1H), 4.02 (s, 2H), 3.89 (s, 1H), 3.80-3.70 (m, 1H), 3.18-2.85
(m, 9H), 2.60-2.50 (m, 1H), 2.32-2.10 (m, 5H), 1.90-1.80 (m, 2H),
1.67-1.52 (m, 1H); MS (ESI) m/z 566.1 (M+H).
[0661] Compound 323. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.55
(s, 1H), 4.04 (s, 1H), 4.02 (s, 2H), 3.60-3.50 (m, 1H), 3.18-2.78
(m, 9H), 2.60-2.50 (m, 1H), 2.21-2.14 (m, 1H), 2.07 (br s, 2H),
1.76 (br s, 2H), 1.61 (br s, 5H); MS (ESI) m/z 580.1 (M+H).
[0662] Compound 324. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.63
(s, 1H), 4.12 (s, 1H), 4.10 (s, 2H), 3.25-2.93 (m, 10H), 2.69-2.59
(m, 1H), 2.28-2.20 (m, 1H), 2.15-2.12 (m, 2H), 1.96-1.85 (m, 2H),
1.75-1.65 (m, 2H), 1.45-1.31 (m, 5H); MS (ESI) m/z 594.1 (M+H).
[0663] Compound 325. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.62
(s, 1H), 4.29 (d, J=16.0 Hz, 1H), 4.18 (d, J=16.0 Hz, 1H), 4.12 (s,
1H), 3.28-2.92 (m, 14H), 2.70-2.58 (m, 1H), 2.28-2.20 (m, 1H),
1.72-1.53 (m, 1H), 1.38 (t, J=7.2 Hz, 3H); MS (ESI) m/z 554.0
(M+H).
[0664] Compound 326. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.60
(s, 1H), 4.24 (s, 2H), 4.11 (s, 1H), 3.38-3.28 (m, 4H), 3.25-2.98
(m, 9H), 2.68-2.58 (m, 1H), 2.28-2.20 (m, 1H), 1.71-1.62 (m, 1H),
1.36 (t, J=7.2 Hz, 6H); MS (ESI) m/z 568.0 (M+H).
[0665] Compound 327. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.62
(s, 1H), 4.33 (d, J=16.0 Hz, 1H), 4.21 (d, J=16.0 Hz, 1H), 4.14 (s,
1H), 3.30-2.88 (m, 14H), 2.70-2.60 (m, 1H), 2.30-2.22 (m, 1H),
1.90-1.77 (m, 2H), 1.75-1.60 (m, 1H), 1.04 (t, J=7.6 Hz, 3 H); MS
(ESI) m/z 568.0 (M+H).
[0666] Compound 328. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.51
(s, 1H), 4.30 (br s, 4H), 4.15 (dd, J=9.6, 18.8 Hz, 2H), 4.08 (s,
1H), 3.30-2.88 (m, 9H), 2.64-2.50 (m, 2H), 2.42 (br s, 1H),
2.24-2.15 (m, 1H), 1.70-1.50 (m, 1H); MS (ESI) m/z 552.0 (M+H).
[0667] Compound 329. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.57
(s, 1H), 4.07 (s, 1H), 4.04 (s, 2H), 3.46-3.41 (m, 1H), 3.23-2.82
(m, 9H), 2.62-2.55 (m, 1H), 2.25-2.20 (m, 1H), 1.65-1.52 (m, 1H),
1.35 (d, J=6.4 Hz, 6H); MS (ESI) m/z 554.0 (M+H).
[0668] Compound 330. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.54
(s, 1H), 5.50-5.38 (m, 1H), 4.35 (br s, 2H), 4.04 (s, 1H),
4.00-3.71 (m, 2H), 3.50-3.28 (m, 2H), 3.29-2.82 (m, 9H), 2.60-2.24
(m, 4H), 1.66-1.50 (m, 1H); MS (ESI) m/z 584.0 (M+H).
Example 14
Antibacterial Activity
[0669] The antibacterial activities for the compounds of the
invention were studied according to the following protocols.
Minimum Inhibitory Concentration (MIC) Assay
[0670] MICs were determined according to the Clinical and
Laboratory Standards
[0671] Institute (CLSI) guidances (e.g., CLSI. Performance
standards for antimicrobial susceptibility testing; nineteenth
information supplement. CLSI document M100-S19, CLSI, 940 West
Valley Road, Suite 1400, Wayne, Pa. 19087-1898, USA, 2009).
Briefly, frozen bacterial strains were thawed and subcultured onto
Mueller Hinton Broth (MHB) or other appropriate media
(Streptococcus requires blood and Haemophilus requires hemin and
NAD). Following incubation overnight, the strains were subcultured
onto Mueller Hinton Agar and again incubated overnight. Colonies
were observed for appropriate colony morphology and lack of
contamination. Isolated colonies were selected to prepare a
starting inoculum equivalent to a 0.5 McFarland standard. The
starting inoculum was diluted 1:125 using MHB for further use. Test
compounds were prepared by dilution in sterile water to a final
concentration of 5.128 mg/mL. Antibiotics (stored frozen, thawed
and used within 3 hours of thawing) and compounds were further
diluted to the desired working concentrations.
[0672] The assays were run as follows. Fifty .mu.L of MHB was added
to wells 2-12 of a 96-well plate. One hundred .mu.L of
appropriately diluted antibiotics was added to well 1. Fifty .mu.L
of antibiotics was removed from well 1 and added to well 2 and the
contents of well 2 mixed by pipetting up and down five times. Fifty
.mu.L of the mixture in well 2 was removed and added to well 3 and
mixed as above. Serial dilutions were continued in the same manner
through well 12. Fifty .mu.L was removed from well 12 so that all
contained 50 .mu.L. Fifty .mu.L of the working inoculum was then
added to all test wells. A growth control well was prepared by
adding 50 .mu.L of working inoculum and 50 .mu.L of MHB to an empty
well. The plates were then incubated at 37.degree. C. overnight,
removed from the incubator and each well was read on a plate
reading mirror. The lowest concentration (MIC) of test compound
that inhibited the growth of the bacteria was recorded.
Example
TABLE-US-00005 [0673] 1 2 3 4 5 6 7 8 9 10 11 12 [Abt] 32 16 8 4 2
1 0.5 0.25 0.125 0.06 0.03 0.015 grow - - - - - + + + + + + + [abt]
= antibiotic concentration in the well Grow = bacterial growth
(cloudiness)
[0674] Interpretation: MIC=2 .mu.g/mL
Protocol for Determining Inoculum Concentration (Viable Count)
[0675] Ninety .mu.l of sterile 0.9% NaCl was pipetted into wells
2-6 of a 96-well microtiter plate. Fifty 50 .mu.l of the inoculum
was pipetted into well 1. Ten .mu.L from was removed from well 1
and added it to well 2 followed by mixing. Ten .mu.L was removed
from well two and mixed with the contents of well 3 and so on
creating serial dilutions through well 6. Ten .mu.L was removed
from each well and spotted onto an appropriate agar plate. The
plate was placed into an incubator overnight. The colonies in spots
that contain distinct colonies were counted. Viable count was
calculated by multiplying the number of colonies by the dilution
factor.
TABLE-US-00006 Spot from Well 1 2 3 4 5 6 Dilution 10.sup.2
10.sup.3 10.sup.4 10.sup.5 10.sup.6 10.sup.7 Factor
Bacterial Strains
[0676] Fifteen bacterial strains, listed below, were examined in
minimum inhibitory concentration (MIC) assays.
TABLE-US-00007 ID Organism Source Resistance Comments Gram Rx SA100
S. aureus ATCC 13709 MSSA Smith strain positive SA101 S. aureus
ATCC 29213 MSSA control positive SA158 S. aureus MR, SK75 tetK
(efflux) positive SA161 S. aureus Micromyx, LLC tet(M) ribosomal
positive protection EF103 E. faecalis ATCC 29212 tet-I/R control
positive EF159 E. faecalis MR, DS160 tet(M) (rib cip-R, ery-I
positive protect) SP106 S. pneumoniae ATCC 49619 wt control
positive SP160 S. pneumoniae MR, 54 tet(M) (rib pen-R, ery-R
positive protect) EC107 E. coli ATCC 25922 wt control negative
EC155 E. coli MR, 10 tet(A) (efflux) negative KP109 K. pneumoniae
ATCC 13883 wt negative KP153 K. pneumoniae MR, 1 tet(A) (efflux)
cip-R, gen-R negative EC108 E. cloacae ATCC 13047 wt negative AB110
A. baumanii ATCC 19606 wt negative PA111 P. aeruginosa ATCC 27853
wt control negative MSSA = methicillin susceptible S. aureus wt =
wild type ATCC = American Type Culture Collection MR = Marilyn
Roberts, University of Washington tet = tetracycline tet-I/R =
tetracycline intermediate/resistant mechanism no specified cip =
ciprofloxacin R = resistant gen = gentamicin ery = erythromycin pen
= penicillin
Results
[0677] Values of minimum inhibition concentration (MIC) for the
compounds of the invention are provided in Table 5.Table 5. MIC
Values for Compounds of the Invention Compared to Sancycline,
Minocycline and Tigecycline. A=lower than or equal to lowest MIC
among three control compounds; B=greater than lowest MIC among
three control compounds, but lower than highest MIC among three
control compounds; C=greater than MIC of all three control
compounds.
TABLE-US-00008 SA161 SA101 SA100 MRSA, SA158 EF103 EF159 SP106
SP160 EC107 EC155 AB110 PA111 EC108 KP109 KP153 Cmpd 29213 13709
tetM tetK 29212 tetM 49619 tetM 25922 tetA 19606 27853 13047 13883
tetA 100 B B B B B B B C B B B C B B B 101 B B A B B B A B B B C B
B B B 102 B B B B B B B A B B C A B B B 103 B B B B B B A A B B B C
B B B 104 B B B B B B A A B B B C B B B 105 B B B B B B A A B B B B
B B B 106 A B B B A B A A B B B B B B B 107 B B B B B B A A B B C B
B B B 108 B B C B B B B B B B C C B B B 109 B B B B B B A A B B C C
B B B 110 C C C C B C C C C C C C C C C 111 C C B B B B C B C C C C
C C C 112 B B B B B B A A B B C C B B B 113 C C B B B B C B C C C C
C C C 114 B A B B B B A A B B C C B B B 115 B B A B B B A A B B B C
B B B 116 B B B B B B C B C C C C C C C 117 C B B B B B C B C C C C
C C C 118 C C B B B B B B B B C C C B B 119 C C C C B B C B C C C C
C C C 120 B B B B B B A B B B C A B B B 121 B B B C B B B B B C C C
B B C 122 C B B C B B B B C C C C C C C 123 B B B B B B A B B B C C
B B B 124 B B B C B B B B B B C C B B B 125 B B B B B B B B B A A B
B B B 126 B A B B A B A A B B C B B B B 127 B B B B B B A A B B B C
B B B 128 B B B B A B A B B B A C B B B 129 B B B B B B B B B B B C
B B B 130 B B B B B B A A B B A C B B B 131 B B B B B B A A B B C C
B B B 132 B B B B A B A A B B B C B B B 133 B B B B B B A A B B B C
B B B 134 C C B B B B C B C C C C C C C 135 B B B B A B A A B B A C
B B B 136 B B B B A B A A B B B C B B B 137 B B B B B B B B B B B C
B B B 138 B B B B B B B B B B B C C B B 139 B B B B B B C C C C C C
C C C 140 C C B C B B C B C C C C C C C 141 C C B B B B C B B C C C
C C C 142 B B B B B B A B B B C B B B B 143 B B B B B B B B B B C C
B B B 144 C B B B B B B A B B C C B B B 145 B B A B B A A A B B B B
B B B 146 A A A A A A A A B B A B A B B 147 A B A B B A A A B B B B
A B B 148 B B A B B A A A B B A C B B B 149 C C B B B B C B B B C C
B B B 150 B B B B B B B B B C C C B B B 151 C B B B B B B B B B C C
B B B 152 B B B B B B A A B B C C B B B 153 B B B B B B C B B C C C
C C C 154 A A A B A B A B B C C C B B C 155 C C C C C C C C C C C C
C C C 156 C B B B B B B B C C C C C C C 157 B B B B B B B B C C C C
C C C 158 C B B B B B B B B C C C C C C 159 C B B B B B B B B C C C
C C C 160 B B B B B B B B C C C C C C C 161 B B B B B B B B C C C C
C C C 162 B C B B B B B B C C C C C C C 163 C C B B B B B B B C C C
C C C 164 C C C C B B C C C C C C C C C 165 B B B C B B C B C C C C
C C C 166 B B B B B B B B B C C C C B C 167 B B B B B B C B B C C C
C C C 168 B B B B B B B B B C C C C C C 169 B A B B B B A B B C C C
B B C 170 B B B B B B A B B C C C C C C 171 B B B B B B B B B C C C
C C C 172 C B B B B B B B C C C C C C C 173 B B B B B B A B B C C C
C C C 174 B B B B B B B B B C C C C C C 175 B B B C B B B B B C C C
B B C 176 B B B B B B B B C C C C C C C 177 C B B B B B C B C C C C
C C C 178 C C C C C C C C C C C C C C C 179 C C B B B B B B B B C C
B B B 180 C C C C C C C C C C C C C C C 181 C C C C C C C C C C C C
C C C 182 C C C C C C C C C C C C C C C 183 C C C C C C C C C C C C
C C C 184 C C C C C C C C C C C C C C C 185 C C C C C C C C C C C C
C C C 186 C C C C C C C C C C C C C C C 187 B B B B B B C B C C C C
C C C 188 C C C C C C C C C C C C C C C 189 C C C C C C C C C C C C
C C C 190 C C C C C C C B C C C C C C C 191 C B B B B B B B B C C C
C B B 192 B B B B B B B B B B C C B B B 193 C C B B B B C B B B C C
C C B 194 C C B B B B C B B B C C C B B 195 C B B B B B B B B C C C
B B C 196 C C C C C B C B C C C C C C C 197 B B B B B B B B B C C C
C B C 198 B B B B B B C B C C C C C C C 199 C C C C B C C B C C C C
C C C 200 C C C C C C C C C C C C C C C 201 C C C C B C C B C C C C
C C C 202 C C C B B B C B C C C C C C C 203 C C B B B B C B C C C C
C C C 204 C C C C B C C B C C C C C C C 205 C C C C C C C C C C C C
C C C 206 C C C C C C C C C C C C C C C 207 B B B B B B B B B B B C
C B B 208 C C C C C C C C C C C C C C C 209 A A B B B B B B B B B B
B B B 210 A A A B A A A A B B B B B B B 211 C B B C B B C B C C C C
C C C 212 A A B B A B A B B C B C B B C 213 B B B B A B A A B B A A
B B B 214 B B B B B B B B B B A C B B B 215 B B B B B B B B B B B C
B B B 216 C C B B B B C B B B C C C B B 217 B B B B B B A A B B B B
B B B 218 A A A B A A A A B B B B B B B 219 B B B B B B C B C C C C
C C C 220 A A A B A A A A B B A B B B B 221 B B B B B B A A B C B C
B B C 222 B B C B B B A B B C C C C B C 223 B B B B B B B B B B B C
B B B 224 A A B B A A A A B B B B B B B 225 B B B B B B B B B B B C
B B B 226 B B B B B B A A B B C C B B B 227 B B B B B B B B B C C C
C C C 228 C C C C B C B B C C C C C C C 229 C C C C C C C C C C C C
C C C 230 B B B B A B A A B B B C B B B 231 B B B B B B C B B B C C
B B B 232 B B B B B B B B B B B C B B B 233 B B B B B B B B B B B C
B B B 234 B B B B B B A B B B B C B B B 235 B B B B B B B B C C C C
C C C 236 B B B B B B B B B B B C B B B 237 B B B B B B B B B B B C
B B B 238 B B B B B B C C C C C C C C C 239 B B B B B B A B B C B C
B B C 240 B B B B B B B B B C C C C C C 241 B B B B B B B B B C C C
C C C 242 B B B B B B B B B B B C C B B 243 B A B B B B B B B B A B
B B B 244 A A A B A A A A B B A B B B NT 245 B B B B A B A A B B B
C B B B 246 B B B B B B A A B B B C B B B 247 B B A B B B C C C C C
C C C C 248 B B A B B B C C C C C C C C C 249 B C B B C B C C C C C
C C C C 250 B B A B B B C C C C C C C C C 251 B A A B B B C C C C C
C C C C 252 B B A B B B C C C C C C C C C 253 B B B B B B C C C C C
C C C C 254 B B B C B B C B C C C C C C C 255 C B B C B B C B C C C
C C C C 256 B B B B B B C C C C C C C C C 257 B A B B A A A A B B C
B B B B 258 A A A B A A A A A B A B B B B 259 B B B B B A B A B B B
C B B B 260 B A B B A A A B B B C A B B B 261 A A B B A A A A B B C
B B B B 262 C C C C C C C C C C C C C C C 263 B B B B B B A A B B A
C B B B 264 B B B B B B C C C C C C C C C 265 C B B B B B C C C C C
C C C C 266 B B B B B B B B B C C C C B C 267 B B B B B B B B B C C
C C C C 268 C C B B B B C C C C C C C C C 269 B B B B B B C B B B B
C C B B 270 A A B B B B A B B B B B B B B 271 B B B B B B A A B B C
C B B B 272 B B B B B B A A B B C C B B B 273 B B B B B B A A B B C
C B B B 274 C B B B B B B B B C C C B B C 275 B B B B B B A B B B C
B B B B 276 C B B B B B B C B B C C B B B 277 C B B B B B B A B B C
B B B B 278 C C B C B B A B B B C C C B B 279 B B B B A B A A B B C
B B B B 280 C B B C B B B B B C C C C B C 281 B B B B B B B B B B A
C B B B 282 B A B B B B B B B B B C B B B 283 B B B B B B B B B B A
C B B B 284 C B B C B B C B C C C C C C C 285 B B B B B B B B B C C
C C B C 286 A A B B A B B B B B B B B B B 287 B B B B B B C B B B B
C B B B 288 C B B B B B B B C C C C C C C 289 B B B B B B B B B B B
C B B B 290 B B B B B B B B B C C C C B C 291 B B B B B B A B B C C
C C B C 292 A A B B A B A A B B B B B B B 293 B A B B A B A A B B A
B B B B 294 A A B C B B A B B B C C B B C 295 A A B B A A B B B B A
B B B B 296 B B B B B B A B B B B C B B B 297 B B B B B B B B B B B
C B B B 298 B B B B B B C B B C C C C C C 299 B A B B A B A A B C C
A B B C 300 C B B C B B C B B C C C C C C 301 B B B B B B B B B B B
C B B B 302 B A B B A B A A B C C C B B C 303 B B B C B B A B B C C
C C B C 304 C B B B B B C C C C C C C C C 305 C C B B B B C C C C C
C C C C 306 C C C C C C C B C C C C C C C 307 B B B B B B A B B C C
B B B C 308 B B B B A A A A B B C B B B B 309 A A A B A A A A B B B
C B B B 310 B B B B B B B B B C C C B B C 311 A B A B A A A A B B A
C B B B 312 A A B B B B A B B C A B B B B 313 B B C C B C C C B C C
C B B C 314 C B B B B B B B B C C C B C C 315 C C B B B B B B B C C
C C C C 316 C B B C B B B B B C C C C C C 317 C B B C B B B B B C C
C B C C 318 C C C C B C B B C C C C C C C 319 C C C C B B C B C C C
C C C C 320 C C C C B B B B B C C C C C C 321 C C C B B B B B B B C
C C B B 322 C C C C B B B B B C C C C C C 323 B B B B B B B B B C C
C B B C 324 C B B B B B B B B C C C C C C 325 C B C B B B A B B C C
C B B C 326 C B C B B B B B B C C C B B C 327 C B B B B B B B B C C
C B B C 328 C C B C B B B B B C C C C C C 329 C C C C B B C B C C C
C C C C 330 C C C C C C C C C C C C C C C 331 C B B B B B B B B C C
C C C C 332 C B C B B B A B B C C C C B B 333 C C C C C C C C C C C
C C C C 334 B B C B B B A B B C C C B B C 335 B B B B B B A B B C C
C B B C Mino- 0.06 0.06 8 0.03 1 16 0.016 2 0.5 8 0.06 16 2 1 8
cycline San- 0.5 1 NT 4 8 8 0.25 8 8 32 0.25 >32 8 8 32 cycline
Tige- 0.06 0.06 0.125 0.06 0.03 0.06 0.016 0.016 0.03 0.5 0.25 8
0.25 0.125 1 cycline
Example 15
In Vivo Activities
[0678] Murine Systemic Infection Model with Intraperitoneal
Challenge of S. aureus
[0679] In the model, CD-1 female mice (18-20 grams) were injected
intraperitoneally with S. aureus ATCC 13709 (Smith)
(1-2.times.10.sup.6/mouse) mixed with 5% hog gastric mucin, a
bacterial inoculum previously established through virulence studies
required to achieve 90-100% lethality within 24-48 hours.
Typically, six mice were treated intravenously per dose group one
hour post-challenge with either compound 101, compound 105,
compound 145, compound 153, compound 166, and compound 218,
tetracycline or tigecycline at doses ranging from 0.05-10 mg/kg.
After 48 hours, percent survival was calculated and the dose
(mg/kg) effecting 50% survival, the protective dose 50%
(PD.sub.50), was reported along as calculated by Probit
analysis.
Results
[0680] The PD.sub.50 values of compound 101, compound 105, compound
145, compound 153, compound 166, and compound 218 and comparators
tigecycline and tetracycline in the S. aureus ATCC 13709 mouse
septicemia model are described in Table 6.
TABLE-US-00009 PD.sub.50 Cmpd # mg/kg, i.v., SA Smith Septicemia
218 1.4 153 >10 166 4.3 145 0.35 101 0.36 105 0.25 Tigecycline
0.97 Tetracycline 1.0
* * * * *