U.S. patent application number 13/829095 was filed with the patent office on 2014-01-23 for inhibitors of histone deacetylase and prodrugs thereof.
This patent application is currently assigned to METHYLGENE INC.. The applicant listed for this patent is MethylGene Inc.. Invention is credited to Alain Ajamian, Robert Deziel.
Application Number | 20140024608 13/829095 |
Document ID | / |
Family ID | 49947050 |
Filed Date | 2014-01-23 |
United States Patent
Application |
20140024608 |
Kind Code |
A1 |
Deziel; Robert ; et
al. |
January 23, 2014 |
INHIBITORS OF HISTONE DEACETYLASE AND PRODRUGS THEREOF
Abstract
The invention relates to the inhibition of histone deacetylase.
The invention provides compounds, prodrugs thereof, and methods for
inhibiting histone deacetylase enzymatic activity. The invention
also provides compositions and methods for treating cell
proliferative diseases and conditions.
Inventors: |
Deziel; Robert;
(Mount-Royal, CA) ; Ajamian; Alain; (Montreal,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MethylGene Inc. |
Montreal |
|
CA |
|
|
Assignee: |
METHYLGENE INC.
Montreal
CA
|
Family ID: |
49947050 |
Appl. No.: |
13/829095 |
Filed: |
March 14, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11959204 |
Dec 18, 2007 |
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13829095 |
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Current U.S.
Class: |
514/25 ; 514/311;
514/352; 514/44A; 514/443; 514/445; 514/603; 514/604; 536/17.6;
546/172; 546/312; 549/55; 549/65; 564/87; 564/92 |
Current CPC
Class: |
C07C 317/44 20130101;
C07D 213/76 20130101; C07C 259/06 20130101; C07D 213/56 20130101;
C07H 15/203 20130101; A61K 31/47 20130101; C07C 311/21 20130101;
C07D 333/62 20130101; A61K 31/18 20130101; A61P 31/10 20180101;
C07D 317/60 20130101; C07D 215/36 20130101; C07D 271/12 20130101;
C07C 275/42 20130101; A61K 31/7088 20130101; C07D 307/46 20130101;
A61K 31/381 20130101; A61K 31/44 20130101; C07D 333/34 20130101;
A61P 35/00 20180101; C07C 323/56 20130101; A61K 31/7028 20130101;
C07D 285/135 20130101 |
Class at
Publication: |
514/25 ; 549/55;
514/443; 514/44.A; 564/87; 514/603; 564/92; 514/604; 549/65;
514/445; 546/172; 514/311; 546/312; 514/352; 536/17.6 |
International
Class: |
A61K 31/7088 20060101
A61K031/7088; A61K 31/381 20060101 A61K031/381; C07C 311/21
20060101 C07C311/21; A61K 31/18 20060101 A61K031/18; A61K 31/7028
20060101 A61K031/7028; C07D 215/36 20060101 C07D215/36; A61K 31/47
20060101 A61K031/47; C07D 213/76 20060101 C07D213/76; A61K 31/44
20060101 A61K031/44; C07H 15/203 20060101 C07H015/203; C07D 333/62
20060101 C07D333/62; C07D 333/34 20060101 C07D333/34 |
Claims
1.-34. (canceled)
35. A compound selected from the group consisting of
N-hydroxy-2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetamide;
2-(4-(4-(benzo[c][1,2,5]oxadiazol-5-yl)but-3-ynyl)phenyl)-N-hydroxyacetam-
ide; N-Hydroxy-2-(3-(4-phenylbutyl)phenyl)acetamide;
N-hydroxy-2-(4-(4-(4-(trifluoromethyl)phenyl)butyl)phenyl)acetamide;
2-(4-(4-(1H-indol-5-yl)butyl)phenyl)-N-hydroxyacetamide;
N-hydroxy-2-(4-(4-(3-(trifluoromethyl)phenyl)butyl)phenyl)acetamide;
N-hydroxy-2-(4-(4-(2-(trifluoromethyl)phenyl)butyl)phenyl)acetamide;
N-hydroxy-2-(4-(4-(imidazo[1,2-a]pyridin-6-yl)butyl)phenyl)acetamide;
2-(4-(4-(benzo[d][1,3]dioxol-5-yl)butyl)phenyl)-N-hydroxyacetamide;
N-hydroxy-2-(4-(5-phenylpentyl)phenyl)acetamide;
N-hydroxy-2-(4-(4-(pyridin-4-yl)butyl)phenyl)acetamide;
N-hydroxy-2-(4-(4-(pyridin-3-yl)butyl)phenyl)acetamide;
N-hydroxy-2-(4-(4-(pyridin-2-yl)butyl)phenyl)acetamide;
N-hydroxy-2-(4-(3-phenylpropyl)phenyl)acetamide; and
N-hydroxy-4-(5-phenylpentyl)benzamide; and pharmaceutically
acceptable salts thereof.
36. A compound selected from the group consisting of ##STR00340##
##STR00341## ##STR00342## and pharmaceutically acceptable salts
thereof.
37. A composition comprising a compound of claim 35 or a
pharmaceutically acceptable salt thereof.
38. A composition comprising a compound of claim 36 or a
pharmaceutically acceptable salt thereof.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of priority of U.S.
Application No. 60/870,768, filed Dec. 19, 2006.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] This invention relates to the inhibition of histone
deacetylase. More particularly, the invention relates to compounds
and prodrugs thereof that inhibit, histone deacetylase enzymatic
activity. The invention also relates to methods of inhibiting
histone deacetylase enzymatic activity.
[0004] 2. Summary of the Related Art
[0005] In eukaryotic cells, nuclear DNA associates with histones to
form a compact complex called chromatin. The histones constitute a
family of basic proteins which are generally highly conserved
across eukaryotic species. The core histones, termed H2A, H.sub.2B,
H3, and H4, associate to form a protein core. DNA winds around this
protein core, with the basic amino acids of the histones
interacting with the negatively charged phosphate groups of the
DNA. Approximately 146 base pairs of DNA wrap around a histone core
to make up a nucleosome particle, the repeating structural motif of
chromatin.
[0006] Csordas, Biochem. J., 286: 23-38 (1990) teaches that
histones are subject to posttranslational acetylation of the
e-amino groups of N-terminal lysine residues, a reaction that is
catalyzed by histone acetyl transferase (HAT1). Acetylation
neutralizes the positive charge of the lysine side chain, and is
thought to impact chromatin structure. Indeed, Taunton et al.,
Science, 272: 408-411 (1996), teaches that access of transcription
factors to chromatin templates is enhanced by histone
hyperacetylation. Taunton et al. further teaches that an enrichment
in underacetylated histone H4 has been found in transcriptionally
silent regions of the genome.
[0007] Histone acetylation is a reversible modification, with
deacetylation being catalyzed by a family of enzymes termed histone
deacetylases (HDACs). Grozinger et al., Proc. Natl. Acad. Sci. USA,
96: 4868-4873 (1999), teaches that HDACs may be divided into two
classes, the first represented by yeast Rpd3-like proteins, and the
second represented by yeast Hda1-like proteins.
[0008] Histone deacetylases play an important role in gene
regulation in mammalian cells. Gray and Ekstrom, Expr. Cell. Res.
262: 75-83 (2001); Zhou et al., Proc. Natl. Acad. Sci. USA 98:
10572-10577 (2001); Kao et al. J. Biol. Chem. 277: 187-193 (2002)
and Gao et al. J. Biol. Chem. 277: 25748-25755 (2002) teach that
there are 11 members of the histone deacetylase (HDAC) family.
Another family of deacetylases involved in gene expression is the
Sir2 family. Gray and Ekstrom, supra, teach that there are seven
members of the Sir2 family in humans.
[0009] Class I human histone deacetylases include HDAC1, HDAC2,
HDAC3 and HDAC8. The Class I enzymes are expressed in a wide
variety of tissues and are reported to be localized in the nucleus.
Class II human histone deacetylases include HDAC4, HDAC5, HDAC6,
HDAC7, HDAC9 and HDAC10. The Class II enzymes have been described
as limited in tissue distribution and they can shuttle between the
nucleus and the cytoplasm. The Class II enzymes are further divided
into Class IIa (HDAC4, HDAC5, HDAC7 and HDAC9) and Class IIb (HDAC6
and HDAC10). Recent classifications place HDAC11 in a class of its
own.
[0010] Richon et al., Proc. Natl. Acad. Sci. USA, 95: 3003-3007
(1998), discloses that HDAC activity is inhibited by trichostatin A
(TSA), a natural product isolated from Streptomyces hygroscopicus,
and by a synthetic compound, suberoylanilide hydroxamic acid
(SAHA). Yoshida and Beppu, Exper. Cell Res., 177: 122-131 (1988),
teaches that TSA causes arrest of rat fibroblasts at the G.sub.1
and G.sub.2 phases of the cell cycle, implicating HDAC in cell
cycle regulation. Indeed, Finnin et al., Nature, 401: 188-193
(1999), teaches that TSA and SAHA inhibit cell growth, induce
terminal differentiation, and prevent the formation of tumors in
mice.
[0011] U.S. Pat. No. RE39850, incorporated herein by reference,
discloses compounds that inhibit HDAC activity for intervening in
cell cycle regulation and therapeutic potential in the treatment of
cell proliferative diseases or conditions. However, these compounds
can exhibit poor bioavailability, thereby limiting their
therapeutic potential. It is therefore desirable to also prepare
bioavailable analogs of such active compounds.
SUMMARY OF THE INVENTION
[0012] The invention provides compounds, prodrugs and methods for
treating diseases or conditions ameliorated by modulating HDAC
activity, such as cell proliferative diseases, or fungal infection,
by administering such prodrugs. In particular, the invention
provides prodrug inhibitors of histone deacetylase enzymatic
activity. These prodrugs are cleavable (e.g., hydrolysable) in a
mammalian cell, a plant cell or fungal pathogen cell. Thus, also
included within the scope of the present invention are products of
the cleaved prodrugs, which include novel hydroxamate based
compounds. For the purpose of clarity, a "prodrug compound" or
"prodrug" of the present invention is intended to mean a
non-cleaved compound as defined by Formula (1), (2) and (3). A
"cleavage product" of the prodrug is intended to mean a prodrug
compound from which the prodrug moiety has been removed.
[0013] In a first aspect, therefore, the invention provides
prodrugs of inhibitors of histone deacetylase, the prodrugs having
the formula (1):
Cy-L.sup.1-Ar-Y.sup.1--C(O)--N(R.sup.x)--Z (1)
and pharmaceutically acceptable salts thereof, wherein [0014] Cy is
--H, cycloalkyl, aryl, heteroaryl, or heterocyclyl, any of which
may be optionally substituted; [0015] L.sup.1 is
--(CH.sub.2).sub.m--W--, where m is 0, 1, 2, 3, or 4, and W is
selected from the group consisting of --C(O)NH--, --S(O).sub.2NH--,
--NHC(O)--, --NHS(O).sub.2--, and --NH--C(O)--NH--; [0016] Ar is
arylene, wherein said arylene optionally may be additionally
substituted and optionally may be fused to an aryl or heteroaryl
ring, or to a saturated or partially unsaturated cycloalkyl or
heterocyclic ring, any of which may be optionally substituted;
[0017] Y.sup.1 is a chemical bond or a straight- or branched-chain
saturated alkylene, wherein said alkylene may be optionally
substituted; [0018] Z is --R.sup.20, --O--R.sup.20, --R.sup.21,
or
##STR00001##
[0018] wherein --R.sup.20 is selected from the group consisting of
--C(O)--R.sup.10, --C(O)O--R.sup.10, --R.sup.11,
--CH(R.sup.12)--O--C(O)--R.sup.10,
--C(O)--C[(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13),
--S(O.sub.2)R.sup.10, --P(O)(OR.sup.10)(OR.sup.10),
--C(O)--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.10,
--C(O)--O--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.10 and
--C(O)--(CH.sub.2).sub.n--C(O)OR.sup.10,
--C(O)--(CH.sub.2).sub.1-4--C(OH)(COOR.sup.10)--(CH.sub.2).sub.1-4--COOR.-
sup.10, --C(O)-[C(R.sup.14)(R.sup.14)].sub.1-4--P(O)(OH)(OH),
--C(O)--(CH.sub.2).sub.1-4--N(R.sup.14)--C[.dbd.N(R.sup.10')]-N(R.sup.10'-
)(R.sup.10'),
--C(O)--(CH.sub.2)--CH(OH)--(CH.sub.2)--N(CH.sub.3)(CH.sub.3),
--C(O)--CH(NH.sub.2)--(CH.sub.2).sub.1-6--COOH (preferably
--C(O)--CH(NH.sub.2)--(CH.sub.2)--COOH), provided that the N to
which Z is bound is not directly bonded to two O atoms; and further
provided that (a) when Z is --R.sup.20 then R.sup.x is --OH, and
(b) when Z is --OR.sup.20 then R.sup.x is --H; [0019] R.sup.x is H
or --OH; [0020] or [0021] R.sup.x is absent and R.sup.20 forms an
optionally substituted heterocyclic ring with the N to which it is
attached; [0022] n is 0, 1, 2, 3, or 4, preferably 1, 2, 3, or 4;
[0023] each R.sup.10 is independently selected from the group
consisting of hydrogen, optionally substituted C.sub.1-C.sub.20
alkyl, optionally substituted C.sub.2-C.sub.20 alkenyl, optionally
substituted C.sub.2-C.sub.20 alkynyl, optionally substituted
C.sub.1-C.sub.20 alkoxycarbonyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, optionally substituted heteroaryl, optionally substituted
cycloalkylalkyl, optionally substituted heterocycloalkylalkyl,
optionally substituted arylalkyl, optionally substituted
heteroarylalkyl, optionally substituted cycloalkylalkenyl,
optionally substituted heterocycloalkylalkenyl, optionally
substituted arylalkenyl, optionally substituted heteroarylalkenyl,
optionally substituted cycloalkylalkynyl, optionally substituted
heterocycloalkylalkynyl, optionally substituted arylalkynyl,
optionally substituted heteroarylalkynyl, optionally substituted
alkylcycloalkyl, optionally substituted alkylheterocycloalkyl,
optionally substituted alkylaryl, optionally substituted
alkylheteroaryl, optionally substituted alkenylcycloalkyl,
optionally substituted alkenylheterocycloalkyl, optionally
substituted alkenylaryl, optionally substituted alkenylheteroaryl,
optionally substituted alkynylcycloalkyl, optionally substituted
alkynylheterocycloalkyl, optionally substituted alkynylary,
optionally substituted alkynylheteroaryl, a sugar residue, and an
amino acid residue (preferably bonded through the carboxy terminus
of the amino acid); [0024] each R.sup.10' is independently hydrogen
or C.sub.1-6alkyl, or [0025] R.sup.10 and R.sup.10' together with
the carbon atom to which they are attached form an optionally
substituted spirocycloalkyl; [0026] R.sup.21 is a sugar or -amino
acid-R.sup.13, wherein R.sup.13 is covalently bound to the
N-terminus; [0027] R.sup.11 is selected from the group consisting
of hydrogen, optionally substituted heterocycloalkyl, optionally
substituted aryl, and optionally substituted heteroaryl; [0028]
R.sup.12 is selected from hydrogen or alkyl; and [0029] R.sup.13 is
selected from the group consisting of hydrogen,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-N(R.sup.10')(R.s-
up.10')C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-aryl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-heteroaryl,
--C(O)-aryl, --C(O)-heteroaryl, an amino protecting group, and
R.sup.10; and [0030] each R.sup.14 is independently selected from
the group consisting of H, C.sub.1-C.sub.6alkyl and cycloalkyl, or
two R.sup.14, together with the atom to which they are attached,
form a cycloalkyl.
[0031] In a second embodiment, the invention provides prodrugs of
inhibitors of histone deacetylase, the prodrugs having the formula
(2):
Cy-L.sup.2-Ar-Y.sup.2--C(O)N(R.sup.x)--Z (2)
and pharmaceutically acceptable salts thereof, wherein [0032] Cy is
H or is cycloalkyl, aryl, heteroaryl, or heterocyclyl, any of which
may be optionally substituted, provided that Cy is not a
(spirocycloalkyl)heterocyclyl; [0033] L.sup.2 is C.sub.1-C.sub.6
saturated alkylene, C.sub.2-C.sub.6 alkenylene or C.sub.2-C.sub.6
alkynylene, wherein the alkylene or alkenylene optionally may be
substituted, and wherein one or two of the carbon atoms of the
alkylene is optionally replaced by a heteroatomic moiety
independently selected from the group consisting of O; NR', R'
being alkyl, acyl, or hydrogen; S; S(O); or S(O).sub.2; [0034] Ar
is arylene, wherein said arylene optionally may be additionally
substituted and optionally may be fused to an aryl or heteroaryl
ring, or to a saturated or partially unsaturated cycloalkyl or
heterocyclic ring, any of which may be optionally substituted;
[0035] Y.sup.2 is a chemical bond or a straight- or branched-chain
saturated alkylene, which may be optionally substituted, provided
that the alkylene is not substituted with a substituent of the
formula --C(O)R wherein R comprises an .alpha.-amino acyl moiety;
[0036] R.sup.x is H or --OH; [0037] Z is --R.sup.20, --O--R.sup.20,
--R.sup.21, or
##STR00002##
[0037] wherein --R.sup.20 is selected from the group consisting of
--C(O)--R.sup.10, --C(O)O--R.sup.10, --R.sup.11,
--CH(R.sup.12)--O--C(O)--R.sup.10,
--C(O)--C[(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13),
--S(O.sub.2)R.sup.10, --P(O)(OR.sup.10)(OR.sup.10),
--C(O)--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.10,
--C(O)--(CH.sub.2).sub.1-4--C(OH)(COOR.sup.10)--(CH.sub.2).sub.1-4--COOR.-
sup.10, --C(O)-[C(R.sup.14)(R.sup.14)].sub.1-4--P(O)(OH)(OH),
--C(O)--(CH.sub.2).sub.1-4--N(R.sup.14)--C[.dbd.N(R.sup.10')]-N(R.sup.10'-
)(R.sup.10'),
--C(O)--(CH.sub.2)--CH(OH)--(CH.sub.2)--N(CH.sub.3)(CH.sub.3),
--C(O)--CH(NH.sub.2)--(CH.sub.2).sub.1-6--COOH (preferably
--C(O)--CH(NH.sub.2)--(CH.sub.2)--COOH),
--C(O)--O--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.10 and
--C(O)--(CH.sub.2).sub.n--C(O)OR.sup.10, provided that the N to
which Z is bound is not directly bonded to two O atoms; and further
provided that (a) when Z is --R.sup.20 then R.sup.x is --OH, and
(b) when Z is --OR.sup.20 then R.sup.x is --H; [0038] or [0039]
R.sup.x is absent and R.sup.20 forms an optionally substituted
heterocyclic ring with the N to which it is attached; [0040] n is
0, 1, 2, 3, or 4, preferably 1, 2, 3, or 4; [0041] each R.sup.10 is
independently selected from the group consisting of hydrogen,
optionally substituted C.sub.1-C.sub.20 alkyl, optionally
substituted C.sub.2-C.sub.20 alkenyl, optionally substituted
C.sub.2-C.sub.20 alkynyl, optionally substituted C.sub.1-C.sub.20
alkoxycarbonyl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, optionally substituted
cycloalkylalkyl, optionally substituted heterocycloalkylalkyl,
optionally substituted arylalkyl, optionally substituted
heteroarylalkyl, optionally substituted cycloalkylalkenyl,
optionally substituted heterocycloalkylalkenyl, optionally
substituted arylalkenyl, optionally substituted heteroarylalkenyl,
optionally substituted cycloalkylalkynyl, optionally substituted
heterocycloalkylalkynyl, optionally substituted arylalkynyl,
optionally substituted heteroarylalkynyl, optionally substituted
alkylcycloalkyl, optionally substituted alkylheterocycloalkyl,
optionally substituted alkylaryl, optionally substituted
alkylheteroaryl, optionally substituted alkenylcycloalkyl,
optionally substituted alkenylheterocycloalkyl, optionally
substituted alkenylaryl, optionally substituted alkenylheteroaryl,
optionally substituted alkynylcycloalkyl, optionally substituted
alkynylheterocycloalkyl, optionally substituted alkynylary,
optionally substituted alkynylheteroaryl, a sugar residue, and an
amino acid residue (preferably bonded through the carboxy terminus
of the amino acid); [0042] each R.sup.10' is independently hydrogen
or C.sub.1-6alkyl, or [0043] R.sup.10 and R.sup.10' together with
the carbon atom to which they are attached form an optionally
substituted spirocycloalkyl; [0044] R.sup.21 is a sugar or -amino
acid-R.sup.13, wherein R.sup.13 is covalently bound to the
N-terminus; [0045] R.sup.11 is selected from the group consisting
of hydrogen, optionally substituted heterocycloalkyl, optionally
substituted aryl, and optionally substituted heteroaryl; [0046]
R.sup.12 is selected from hydrogen or alkyl; and [0047] R.sup.13 is
selected from the group consisting of hydrogen,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-N(R.sup.10')(R.s-
up.10'),
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-aryl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-heteroaryl,
--C(O)-aryl, --C(O)-heteroaryl, an amino protecting group, and
R.sup.10; and [0048] each R.sup.14 is independently selected from
the group consisting of H, C.sub.1-C.sub.6alkyl and cycloalkyl, or
two R.sup.14, together with the atom to which they are attached,
form a cycloalkyl.
[0049] In a third embodiment, the invention provides prodrugs of
inhibitors of histone deacetylase, the prodrugs having the formula
(3):
Cy-L.sup.3-Ar-Y.sup.3--C(O)N(R.sup.x)--Z (3)
[0050] and pharmaceutically acceptable salts thereof, wherein
[0051] Cy is --H, cycloalkyl, aryl, heteroaryl, or heterocyclyl,
any of which may be optionally substituted, provided that Cy is not
a (spirocycloalkyl)heterocyclyl; [0052] L.sup.3 is selected from
the group consisting of [0053] (a) --(CH.sub.2).sub.m--W--, where m
is 0, 1, 2, 3, or 4, and W is selected from the group consisting of
--C(O)NH--, --S(O).sub.2NH--, --NHC(O)--, --NHS(O).sub.2--, and
--NH--C(O)--NH--; and [0054] (b) C.sub.1-C.sub.6 alkylene or
C.sub.2-C.sub.6 alkenylene, wherein the alkylene or alkenylene
optionally may be substituted, and wherein one of the carbon atoms
of the alkylene optionally may be replaced by O; NR', R' being
alkyl, acyl, or hydrogen; S; S(O); or S(O).sub.2; [0055] Ar is
arylene, wherein said arylene optionally may be additionally
substituted and optionally may be fused to an aryl or heteroaryl
ring, or to a saturated or partially unsaturated cycloalkyl or
heterocyclic ring, any of which may be optionally substituted; and
[0056] Y.sup.3 is C.sub.2 alkenylene or C.sub.2 alkynylene, wherein
one or both carbon atoms of the alkenylene optionally may be
substituted with alkyl, aryl, alkaryl, or aralkyl; [0057] R.sup.x
is H or --OH; [0058] Z is --R.sup.20, --O--R.sup.20, --R.sup.21,
or
##STR00003##
[0058] wherein --R.sup.20 is selected from the group consisting of
--C(O)--R.sup.10, --C(O)O--R.sup.10, --R.sup.11,
--CH(R.sup.12)--O--C(O)--R.sup.10,
--C(O)--C[(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13),
--S(O.sub.2)R.sup.10--C(O)--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.-
10,
--C(O)--(CH.sub.2).sub.1-4--C(OH)(COOR.sup.10)--(CH.sub.2).sub.1-4--CO-
OR.sup.10, --C(O)-[C(R.sup.14)(R.sup.14)].sub.1-4--P(O)(OH)(OH),
--C(O)--(CH.sub.2).sub.1-4--N(R.sup.14)--C[.dbd.N(R.sup.10')]-N(R.sup.10'-
)(R.sup.10'),
--C(O)--(CH.sub.2)--CH(OH)--(CH.sub.2)--N(CH.sub.3)(CH.sub.3),
--C(O)--CH(NH.sub.2)--(CH.sub.2).sub.1-6--COOH (preferably
--C(O)--CH(NH.sub.2)--(CH.sub.2)--COOH),
--C(O)--O--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.10 and
--C(O)--(CH.sub.2).sub.n--C(O)OR.sup.10, provided that the N to
which Z is bound is not directly bonded to two O atoms; and further
provided that (a) when Z is --R.sup.20 then R.sup.x is --OH, and
(b) when Z is --OR.sup.20 then R.sup.x is --H; [0059] or [0060]
R.sup.x is absent and R.sup.20 forms an optionally substituted
heterocyclic ring with the N to which it is attached; [0061] n is
0, 1, 2, or 4, preferably 1, 2, 3, or 4; [0062] each R.sup.10 is
independently selected from the group consisting of hydrogen,
optionally substituted C.sub.1-C.sub.20 alkyl, optionally
substituted C.sub.2-C.sub.20 alkenyl, optionally substituted
C.sub.2-C.sub.20 alkynyl, optionally substituted C.sub.1-C.sub.20
alkoxycarbonyl, optionally substituted cycloalkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl,
optionally substituted heteroaryl, optionally substituted
cycloalkylalkyl, optionally substituted heterocycloalkylalkyl,
optionally substituted arylalkyl, optionally substituted
heteroarylalkyl, optionally substituted cycloalkylalkenyl,
optionally substituted heterocycloalkylalkenyl, optionally
substituted arylalkenyl, optionally substituted heteroarylalkenyl,
optionally substituted cycloalkylalkynyl, optionally substituted
heterocycloalkylalkynyl, optionally substituted arylalkynyl,
optionally substituted heteroarylalkynyl, optionally substituted
alkylcycloalkyl, optionally substituted alkylheterocycloalkyl,
optionally substituted alkylaryl, optionally substituted
alkylheteroaryl, optionally substituted alkenylcycloalkyl,
optionally substituted alkenylheterocycloalkyl, optionally
substituted alkenylaryl, optionally substituted alkenylheteroaryl,
optionally substituted alkynylcycloalkyl, optionally substituted
alkynylheterocycloalkyl, optionally substituted alkynylary,
optionally substituted alkynylheteroaryl, a sugar residue, and an
amino acid residue (preferably bonded through the carboxy terminus
of the amino acid); [0063] each R.sup.10' is independently hydrogen
or C.sub.1-6alkyl, or [0064] R.sup.10 and R.sup.10' together with
the carbon atom to which they are attached form an optionally
substituted spirocycloalkyl; [0065] R.sup.21 is a sugar or -amino
acid-R.sup.13, wherein R.sup.13 is covalently bound to the
N-terminus; [0066] R.sup.11 is selected from the group consisting
of hydrogen, optionally substituted heterocycloalkyl, optionally
substituted aryl, and optionally substituted heteroaryl; [0067]
R.sup.12 is selected from hydrogen or alkyl; and [0068] R.sup.13 is
selected from the group consisting of hydrogen,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-N(R.sup.10')(R.s-
up.10')C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-aryl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-heteroaryl,
--C(O)-aryl, --C(O)-heteroaryl, an amino protecting group, and
R.sup.10; and [0069] each R.sup.14 is independently selected from
the group consisting of H, C.sub.1-C.sub.6alkyl and cycloalkyl, or
two R.sup.14, together with the atom to which they are attached,
form a cycloalkyl.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0070] The invention provides prodrugs, cleavage products thereof,
and methods for inhibiting histone deacetylase enzymatic activity.
The invention also provides compositions and methods for treating
diseases or conditions ameliorated by modulating HDAC activity,
such as cell proliferative diseases and conditions, and fungal
infection. The patent and scientific literature referred to herein
establishes knowledge that is available to those with skill in the
art. The issued patents, applications, and references that are
cited herein are hereby incorporated by reference to the same
extent as if each was specifically and individually indicated to be
incorporated by reference. In the case of inconsistencies, the
present disclosure will prevail.
[0071] For purposes of the present invention, the following
definitions will be used:
[0072] Unless otherwise indicated by context, the terms "histone
deacetylase" and "HDAC" are intended to refer to any one of a
family of enzymes that remove acetyl groups from the
.epsilon.-amino groups of lysine residues at the N-terminus of a
histone. Unless otherwise indicated by context, the term "histone"
is meant to refer to any histone protein, including H1, H2A, H2B,
H3, H4, and H5, from any species. Preferred histone deacetylases
include class I and class II enzymes. Preferably the histone
deacetylase is a human HDAC, including, but not limited to, HDAC-1,
HDAC-2, HDAC-3, HDAC-4, HDAC-5, HDAC-6, HDAC-7, HDAC-8, HDAC-9,
HDAC-10, and HDAC-11. In some other preferred embodiments, the
histone deacetylase is derived from a protozoal or fungal source.
Preferred fungi include, but are not limited to Saccharomyces
cerevisiae, Candida spp. (such as C. albicans, C. glabrata, C.
tropicalis, C. parapsilosis, C. krusei, C. lusitaniae, C.
dubliniensis), Aspergillus spp. (such as A. fumigatus, A. flavus,
A. niger, A. terreus), Fusarium spp., Paecilomyces lilacinus,
Rhizopus arrhizus and Coccidioides immitis. In certain preferred
embodiments, the histone deacetylase is a fungal HDAC including,
but not limited to Rpd3, Hos1, Hos2, Hda1, Hos3, Sir2, Hst, and
homologs thereof. In preferred embodiments, a cleavage product of a
prodrug compound of the present invention shows synergistic
activity with an antifungal agent against a fungal species,
preferably at concentrations of inhibitor not toxic to mammalian
cells. Preferably such antifungal agents are azole antifungal
agents (a large number of active antifungal agents have an azole
functionality as part of their structure; such an antifungal agent
is generally referred to as an "antifungal azole", an "azole
antifungal agent" or an "azole"). Such combinations, and
compositions thereof, can be used to selectively treat fungal
infection.
[0073] The term "antifungal agent" is intended to mean a substance
capable of inhibiting or preventing the growth, viability and/or
reproduction of a fungal cell. Preferable antifungal agents are
those capable of preventing or treating a fungal infection in an
animal or plant. A preferable antifungal agent is a broad spectrum
antifungal agent. However, an antifungal agent can also be specific
to one or more particular species of fungus.
[0074] Preferred antifungal agents are ergosterol synthesis
inhibitors, and include, but are not limited to azoles and
phenpropimorph. Other antifungal agents include, but are not
limited to terbinafine. Preferred azoles include imidazoles and
triazoles. Further preferred antifungal agents include, but are not
limited to, ketoconazole, itroconazole, fluconazole, voriconazole,
posaconazole, ravuconazole and miconazole. Like azoles,
phenpropimorph is an ergosterol synthesis inhibitor, but acts on
the ergosterol reductase (ERG24) step of the synthesis pathway.
Terbinafine, is also an ergosterol inhibitor, but acts on the
squalene eposidase (ERG1) step.
[0075] The term "histone deacetylase inhibitor" or "inhibitor of
histone deacetylase" is used to identify a compound having a
structure as defined herein, which is capable of interacting with a
histone deacetylase and inhibiting its enzymatic activity.
Inhibiting histone deacetylase enzymatic activity means reducing
the ability of a histone deacetylase to remove an acetyl group from
a histone. In some preferred embodiments, such reduction of histone
deacetylase activity is at least about 50%, more preferably at
least about 75%, and still more preferably at least about 90%. In
other preferred embodiments, histone deacetylase activity is
reduced by at least 95% and more preferably by at least 99%.
[0076] Preferably, such inhibition is specific, i.e., the histone
deacetylase inhibitor reduces the ability of a histone deacetylase
to remove an acetyl group from a histone at a concentration that is
lower than the concentration of the inhibitor that is required to
produce another, unrelated biological effect. Preferably, the
concentration of the inhibitor required for histone deacetylase
inhibitory activity is at least 2-fold lower, more preferably at
least 5-fold lower, even more preferably at least 10-fold lower,
and most preferably at least 20-fold lower than the concentration
required to produce an unrelated biological effect. In certain
preferred embodiments of the present invention, cleavage (e.g.,
hydrolysis) of the prodrug releases a compound (a cleavage (e.g.,
hydrolyzation) product) which is an inhibitor of histone
deacetylase that is more active against a fungal histone
deacetylase than against a mammalian histone deacetylase. In
certain preferred embodiments of the present invention, the
inhibitor of histone deacetylase is specific for a fungal histone
deacetylase.
[0077] The terms "treating", "treatment", or the like, as used
herein covers the treatment of a disease-state in an animal and
includes at least one of: (i) preventing the disease-state from
occurring, in particular, when such animal is predisposed to the
disease-state but has not yet developed symptoms of having it; (ii)
inhibiting the disease-state, i.e., partially or completely
arresting its development; (iii) relieving the disease-state, i.e.,
causing regression of symptoms of the disease-state, or
ameliorating a symptom of the disease; and (iv) reversal or
regression of the disease-state, preferably eliminating or curing
of the disease. In a preferred embodiment the terms "treating",
"treatment", or the like, covers the treatment of a disease-state
in an animal and includes at least one of (ii), (iii) and (iv)
above. In a preferred embodiment of the present invention the
animal is a mammal, preferably a primate, more preferably a human.
As is known in the art, adjustments for systemic versus localized
delivery, age, body weight, general health, sex, diet, time of
administration, drug interaction and the severity of the condition
may be necessary, and will be ascertainable with routine
experimentation by one of ordinary skill in the art.
[0078] For simplicity, chemical moieties are defined and referred
to throughout primarily as univalent chemical moieties (e.g.,
alkyl, aryl, etc.). Nevertheless, such terms are also used to
convey corresponding multivalent moieties under the appropriate
structural circumstances clear to those skilled in the art. For
example, while an "alkyl" moiety generally refers to a monovalent
radical (e.g. CH.sub.3--CH.sub.2--), in certain circumstances a
bivalent linking moiety can be "alkyl," in which case those skilled
in the art will understand the alkyl to be a divalent radical
(e.g., --CH.sub.2--CH.sub.2--), which is equivalent to the term
"alkylene." (Similarly, in circumstances in which a divalent moiety
is required and is stated as being "aryl," those skilled in the art
will understand that the term "aryl" refers to the corresponding
divalent moiety, arylene). All atoms are understood to have their
normal number of valences for bond formation (i.e., 4 for carbon, 3
for N, 2 for O, and 2, 4, or 6 for S, depending on the oxidation
state of the S). On occasion a moiety may be defined, for example,
as (A).sub.a-B--, wherein a is 0 or 1. In such instances, when a is
0 the moiety is B- and when a is 1 the moiety is A-B--.
[0079] For simplicity, reference to a "C.sub.n-C.sub.m,"
heterocyclyl or "C.sub.n-C.sub.m," heteroaryl means a heterocyclyl
or heteroaryl having from "n" to "m" annular atoms, where "n" and
"m" are integers. Thus, for example, a C.sub.5-C.sub.6-heterocyclyl
is a 5- or 6-membered ring having at least one heteroatom, and
includes pyrrolidinyl (C.sub.5) and piperidinyl (C.sub.6);
C.sub.6-heteroaryl includes, for example, pyridyl and
pyrimidyl.
[0080] The term "hydrocarbyl" refers to a straight, branched, or
cyclic alkyl, alkenyl, or alkynyl, each as defined herein. A
"C.sub.0" hydrocarbyl is used to refer to a covalent bond. Thus,
"C.sub.0-C.sub.3-hydrocarbyl" includes a covalent bond, methyl,
ethyl, ethenyl, ethynyl, propyl, propenyl, propynyl, and
cyclopropyl.
[0081] The term "alkyl" is intended to mean a straight or branched
chain aliphatic group having from 1 to 12 carbon atoms, preferably
1-8 carbon atoms, and more preferably 1-6 carbon atoms. Other
preferred alkyl groups have from 2 to 12 carbon atoms, preferably
2-8 carbon atoms and more preferably 2-6 carbon atoms. Preferred
alkyl groups include, without limitation, methyl, ethyl, propyl,
isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and
hexyl. A "C.sub.0" alkyl (as in "C.sub.0-C.sub.3-alkyl") is a
covalent bond.
[0082] The term "alkenyl" is intended to mean an unsaturated
straight or branched chain aliphatic group with one or more
carbon-carbon double bonds, having from 2 to 12 carbon atoms,
preferably 2-8 carbon atoms, and more preferably 2-6 carbon atoms.
Preferred alkenyl groups include, without limitation, ethenyl,
propenyl, butenyl, pentenyl, and hexenyl.
[0083] The term "alkynyl" is intended to mean an unsaturated
straight or branched chain aliphatic group with one or more
carbon-carbon triple bonds, having from 2 to 12 carbon atoms,
preferably 2-8 carbon atoms, and more preferably 2-6 carbon atoms.
Preferred alkynyl groups include, without limitation, ethynyl,
propynyl, butynyl, pentynyl, and hexynyl.
[0084] The terms "alkylene," "alkenylene," or "alkynylene" as used
herein are intended to mean an alkyl, alkenyl, or alkynyl group,
respectively, as defined hereinabove, that is positioned between
and serves to connect two other chemical groups. Preferred alkylene
groups include, without limitation, methylene, ethylene, propylene,
and butylene. Preferred alkenylene groups include, without
limitation, ethenylene, propenylene, and butenylene. Preferred
alkynylene groups include, without limitation, ethynylene,
propynylene, and butynylene.
[0085] The term "cycloalkyl" is intended to mean a saturated or
unsaturated mono-, bi, tri- or poly-cyclic hydrocarbon group having
about 3 to 15 carbons, preferably having 3 to 12 carbons,
preferably 3 to 8 carbons, and more preferably 3 to 6 carbons. In
certain preferred embodiments, the cycloalkyl group is fused to an
aryl, heteroaryl or heterocyclic group. Preferred cycloalkyl groups
include, without limitation, cyclopenten-2-enone,
cyclopenten-2-enol, cyclohex-2-enone, cyclohex-2-enol, cyclopropyl,
cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl,
cycloheptyl, and cyclooctyl.
[0086] The term "heteroalkyl" is intended to mean a saturated or
unsaturated, straight or branched chain aliphatic group, wherein
one or more carbon atoms in the chain are independently replaced by
a heteroatom selected from the group consisting of O, S(O).sub.0-2,
N and N(R.sup.33).
[0087] The term "aryl" is intended to mean a mono-, bi-, tri- or
polycyclic C.sub.6-C.sub.14 aromatic moiety, preferably comprising
one to three aromatic rings. Preferably, the aryl group is a
C.sub.6-C.sub.10 aryl group, more preferably a C.sub.6 aryl group.
Preferred aryl groups include, without limitation, phenyl,
naphthyl, anthracenyl, and fluorenyl.
[0088] The terms "aralkyl" or "arylalkyl" is intended to mean a
group comprising an aryl group covalently linked to an alkyl group.
If an aralkyl group is described as "optionally substituted", it is
intended that either or both of the aryl and alkyl moieties may
independently be optionally substituted or unsubstituted.
Preferably, the aralkyl group is
(C.sub.1-C.sub.6)alk(C.sub.6-C.sub.10)aryl, including, without
limitation, benzyl, phenethyl, and naphthylmethyl. For simplicity,
when written as "arylalkyl" this term, and terms related thereto,
is intended to indicate the order of groups in a compound as
"aryl-alkyl". Similarly, "alkyl-aryl" is intended to indicate the
order of the groups in a compound as "alkyl-aryl".
[0089] The terms "heterocyclyl", "heterocyclic" or "heterocycle"
are intended to mean a group which is a mono-, bi-, or polycyclic
structure having from about 3 to about 14 atoms, wherein one or
more atoms are independently selected from the group consisting of
N, O, and S. The ring structure may be saturated, unsaturated or
partially unsaturated. In certain preferred embodiments, the
heterocyclic group is non-aromatic. In a bicyclic or polycyclic
structure, one or more rings may be aromatic; for example one ring
of a bicyclic heterocycle or one or two rings of a tricyclic
heterocycle may be aromatic, as in indan and 9,10-dihydro
anthracene. Preferred heterocyclic groups include, without
limitation, epoxy, aziridinyl, tetrahydrofuranyl, pyrrolidinyl,
piperidinyl, piperazinyl, thiazolidinyl, oxazolidinyl,
oxazolidinonyl, and morpholino. In certain preferred embodiments,
the heterocyclic group is fused to an aryl, heteroaryl, or
cycloalkyl group. Examples of such fused heterocycles include,
without limitation, tetrahydroquinoline and dihydrobenzofuran.
Specifically excluded from the scope of this term are compounds
where an annular O or S atom is adjacent to another O or S
atom.
[0090] In certain preferred embodiments, the heterocyclic group is
a heteroaryl group. As used herein, the term "heteroaryl" is
intended to mean a mono-, bi-, tri- or polycyclic group having 5 to
14 ring atoms, preferably 5, 6, 9, or 10 ring atoms; having 6, 10,
or 14 pi electrons shared in a cyclic array; and having, in
addition to carbon atoms, between one or more heteroatoms
independently selected from the group consisting of N, O, and S.
For example, a heteroaryl group may be pyrimidinyl, pyridinyl,
benzimidazolyl, thienyl, benzothiazolyl, benzofuranyl and
indolinyl. Preferred heteroaryl groups include, without limitation,
thienyl, benzothienyl, furyl, benzofuryl, dibenzofuryl, pyrrolyl,
imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, indolyl,
quinolyl, isoquinolyl, quinoxalinyl, tetrazolyl, oxazolyl,
thiazolyl, and isoxazolyl.
[0091] The terms "arylene," "heteroarylene," or "heterocyclylene"
are intended to mean an aryl, heteroaryl, or heterocyclyl group,
respectively, as defined hereinabove, that is positioned between
and serves to connect two other chemical groups.
[0092] Preferred heterocyclyls and heteroaryls include, but are not
limited to, acridinyl, azocinyl, benzimidazolyl, benzofuranyl,
benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl,
benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl,
benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl,
chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl,
2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran,
furanyl, furyl, furazanyl, imidazolidinyl, imidazolinyl,
imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl,
indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl, isoindazolyl,
isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl,
methylenedioxyphenyl, morpholinyl, naphthyridinyl,
octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl,
1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl,
oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl,
phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl,
phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl,
4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl,
pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole,
pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl,
pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl,
quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,
tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,
tetrazolyl, 6H-1,2,5-thiadiazinyl, thiadiazolyl (e.g.,
1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl,
1,3,4-thiadiazolyl), thianthrenyl, thiazolyl, thienyl,
thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl,
triazinyl, triazolyl (e.g., 1,2,3-triazolyl, 1,2,4-triazolyl,
1,2,5-triazolyl, 1,3,4-triazolyl), and xanthenyl.
[0093] Aromatic polycycles include, but are not limited to,
bicyclic and tricyclic fused ring systems, including for example
naphthyl.
[0094] Non-aromatic polycycles include, but are not limited to,
bicyclic and tricyclic fused ring systems where each ring can be
4-9 membered and each ring can containing zero, 1 or more double
and/or triple bonds. Suitable examples of non-aromatic polycycles
include, but are not limited to, decalin, octahydroindene,
perhydrobenzocycloheptene and perhydrobenzo-M-azulene.
[0095] Polyheteroaryl groups include bicyclic and tricyclic fused
rings systems where each ring can independently be 5 or 6 membered
and contain one or more heteroatom, for example, 1, 2, 3 or 4
heteroatoms, independently chosen from O, N and S such that the
fused ring system is aromatic. Suitable examples of polyheteroaryl
ring systems include quinoline, isoquinoline, pyridopyrazine,
pyrrolopyridine, furopyridine, indole, benzofuran, benzothiofuran,
benzindole, benzoxazole, pyrroloquinoline, and the like.
[0096] Non-aromatic polyheterocyclic groups include but are not
limited to bicyclic and tricyclic ring systems where each ring can
be 4-9 membered, contain one or more heteratom, for example 1, 2, 3
or 4 heteratoms, independently chosen from O, N and S, and contain
zero, or one or more C--C double or triple bonds. Suitable examples
of non-aromatic polyheterocycles include but are not limited to,
hexitol, cis-perhydro-cyclohepta[b]pyridinyl,
decahydro-benzo[f][1,4]oxazepinyl, 2,8-dioxabicyclo[3.3.0]octane,
hexahydro-thieno[3,2-b]thiophene, perhydropyrrolo[3,2-b]pyrrole,
perhydronaphthyridine, perhydrop-1H-dicyclopenta[b,e]pyran.
[0097] Mixed aryl and non-aryl polyheterocycle groups include but
are not limited to bicyclic and tricyclic fused ring systems where
each ring can be 4-9 membered, contain one or more heteroatom
independently chosen from O, N and S and at least one of the rings
must be aromatic. Suitable examples of mixed aryl and non-aryl
polyheteorcycles include 2,3-dihydroindole,
1,2,3,4-tetrahydroquinoline,
5,11-dihydro-10H-dibenz[b,e][1,4]diazepine,
5H-dibenzo[b,e][1,4]diazepine,
1,2-dihydropyrrolo[3,4-b][1,5]benzodiazepine,
1,5-dihydropyrido[2,3-b][1,4]diazepin-4-one,
1,2,3,4,6,11-hexhydro-benzo[b]pyrido[2,3-e][1,4]diazepine-5-one,
methylenedioxyphenyl, bis-methylenedioxyphenyl,
1,2,3,4-tetrahydronaphthalene, dibenzosuberane dihydroanthracene
and 9H-fluorene.
[0098] As employed herein, and unless stated otherwise, when a
moiety (e.g., alkyl, heteroalkyl, cycloalkyl, aryl, heteroaryl,
heterocyclyl, etc.) is described as "optionally substituted" it is
meant that the group optionally has from one to four, preferably
from one to three, more preferably one or two, non-hydrogen
substituents. Suitable substituents include, without limitation,
halo, hydroxy, oxo (e.g., an annular --CH-- substituted with oxo is
--C(O)--) nitro, halohydrocarbyl, hydrocarbyl, alkyl, cycloalkyl,
heterocyclyl, aryl, heteroaryl, aralkyl, alkoxy, aryloxy, amino,
acylamino, alkylcarbamoyl, arylcarbamoyl, aminoalkyl, acyl,
carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl,
alkanesulfonamido, arenesulfonamido, aralkylsulfonamido,
alkylcarbonyl, acyloxy, cyano, and ureido groups. Preferred
substituents, which are themselves not further substituted (unless
expressly stated otherwise) are: [0099] (a) halo, cyano, oxo,
carboxy, formyl, nitro, amino, amidino, guanidino, [0100] (b)
C.sub.1-C.sub.5 alkyl or alkenyl or arylalkyl imino, carbamoyl,
azido, carboxamido, mercapto, hydroxy, hydroxyalkyl, alkylaryl,
arylalkyl, C.sub.1-C.sub.8 alkyl, C.sub.1-C.sub.8 alkenyl,
C.sub.1-C.sub.8 alkoxy, C.sub.1-C.sub.8 alkoxycarbonyl,
aryloxycarbonyl, C.sub.2-C.sub.8 acyl, C.sub.2-C.sub.8 acylamino,
C.sub.1-C.sub.8 alkylthio, arylalkylthio, arylthio, C.sub.1-C.sub.8
alkylsulfinyl, arylalkylsulfinyl, arylsulfinyl, C.sub.1-C.sub.8
alkylsulfonyl, arylalkylsulfonyl, arylsulfonyl, C.sub.0-C.sub.6
N-alkyl carbamoyl, C.sub.2-C.sub.15 N,N-dialkylcarbamoyl,
C.sub.3-C.sub.7 cycloalkyl, aroyl, aryloxy, arylalkyl ether, aryl,
aryl fused to a cycloalkyl or heterocycle or another aryl ring,
C.sub.3-C.sub.7 heterocycle, C.sub.5-C.sub.15 heteroaryl or any of
these rings fused or spiro-fused to a cycloalkyl, heterocyclyl, or
aryl, wherein each of the foregoing is further optionally
substituted with one more moieties listed in (a), above; and [0101]
(c) --(CR.sup.32R.sup.33a).sub.s--NR.sup.30R.sup.31, wherein s is
from 0 (in which case the nitrogen is directly bonded to the moiety
that is substituted) to 6, R.sup.32 and R.sup.33a are each
independently hydrogen, halo, hydroxyl or C.sub.1-C.sub.4alkyl, and
R.sup.30 and R.sup.31 are each independently hydrogen, cyano, oxo,
hydroxyl, --C.sub.1-C.sub.8 alkyl, C.sub.1-C.sub.8 heteroalkyl,
C.sub.1-C.sub.8 alkenyl, carboxamido, C.sub.1-C.sub.3
alkyl-carboxamido, carboxamido-C.sub.1-C.sub.3 alkyl, amidino,
C.sub.2-C.sub.8hydroxyalkyl, C.sub.1-C.sub.3 alkylaryl,
aryl-C.sub.1-C.sub.3 alkyl, C.sub.1-C.sub.3 alkylheteroaryl,
heteroaryl-C.sub.1-C.sub.3 alkyl, C.sub.1-C.sub.3
alkylheterocyclyl, heterocyclyl-C.sub.1-C.sub.3 alkyl
C.sub.1-C.sub.3 alkylcycloalkyl, cycloalkyl-C.sub.1-C.sub.3 alkyl,
C.sub.2-C.sub.8 alkoxy, C.sub.2-C.sub.8
alkoxy-C.sub.1-C.sub.4alkyl, C.sub.1-C.sub.8 alkoxycarbonyl,
aryloxycarbonyl, aryl-C.sub.1-C.sub.3 alkoxycarbonyl,
heteroaryloxycarbonyl, heteroaryl-C.sub.1-C.sub.3 alkoxycarbonyl,
C.sub.1-C.sub.8 acyl, C.sub.0-C.sub.8 alkyl-carbonyl,
aryl-C.sub.0-C.sub.8 alkyl-carbonyl, heteroaryl-C.sub.0-C.sub.8
alkyl-carbonyl, cycloalkyl-C.sub.0-C.sub.8 alkyl-carbonyl,
C.sub.0-C.sub.8 alkyl-NH-carbonyl, aryl-C.sub.0-C.sub.8
alkyl-NH-carbonyl, heteroaryl-C.sub.0-C.sub.8 alkyl-NH-carbonyl,
cycloalkyl-C.sub.0-C.sub.8 alkyl-NH-carbonyl, C.sub.0-C.sub.8
alkyl-O-carbonyl, aryl-C.sub.0-C.sub.8 alkyl-O-carbonyl,
heteroaryl-C.sub.0-C.sub.8 alkyl-O-carbonyl,
cycloalkyl-C.sub.0-C.sub.8 alkyl-O-carbonyl, C.sub.1-C.sub.8
alkylsulfonyl, arylalkylsulfonyl, arylsulfonyl,
heteroarylalkylsulfonyl, heteroarylsulfonyl, C.sub.1-C.sub.8
alkyl-NH-sulfonyl, arylalkyl-NH-sulfonyl, aryl-NH-sulfonyl,
heteroarylalkyl-NH-sulfonyl, heteroaryl-NH-sulfonyl aroyl, aryl,
cycloalkyl, heterocyclyl, heteroaryl, aryl-C.sub.1-C.sub.3 alkyl-,
cycloalkyl-C.sub.1-C.sub.3 alkyl-, heterocyclyl-C.sub.1-C.sub.3
alkyl-, heteroaryl-C.sub.1-C.sub.3 alkyl-, or protecting group,
wherein each of the foregoing is further optionally substituted
with one more moieties listed in (a), above; or
[0102] R.sup.30 and R.sup.31 taken together with the N to which
they are attached form a heterocyclyl or heteroaryl, each of which
is optionally substituted with from 1 to 3 substituents selected
from the group consisting of (a) above, a protecting group, and
(X.sup.30--Y.sup.31--), wherein said heterocyclyl may also be
bridged (forming a bicyclic moiety with a methylene, ethylene or
propylene bridge); wherein
[0103] X.sup.30 is selected from the group consisting of
C.sub.1-C.sub.8alkyl, C.sub.2-C.sub.8alkenyl-,
C.sub.2-C.sub.8alkynyl-,
--C.sub.0-C.sub.3alkyl-C.sub.2-C.sub.8alkenyl-C.sub.0-C.sub.3alkyl,
C.sub.0-C.sub.3alkyl-C.sub.2-C.sub.8alkynyl-C.sub.0-C.sub.3alkyl,
C.sub.0-C.sub.3alkyl-O--C.sub.0-C.sub.3alkyl-,
HO--C.sub.0-C.sub.3alkyl-,
C.sub.0-C.sub.4alkyl-N(R.sup.30)--C.sub.0-C.sub.3alkyl-,
N(R.sup.30)(R.sup.31)--C.sub.0-C.sub.3alkyl-,
N(R.sup.30)(R.sup.31)--C.sub.0-C.sub.3alkenyl-,
N(R.sup.30)(R.sup.31)--C.sub.0-C.sub.3alkynyl-,
(N(R.sup.30)(R.sup.31)).sub.2--C.dbd.N--,
C.sub.0-C.sub.3alkyl-S(O).sub.0-2--C.sub.0-C.sub.3alkyl-,
CF.sub.3--C.sub.0-C.sub.3alkyl-, C.sub.1-C.sub.8heteroalkyl, aryl,
cycloalkyl, heterocyclyl, heteroaryl, aryl-C.sub.1-C.sub.3alkyl-,
cycloalkyl-C.sub.1-C.sub.3alkyl-,
heterocyclyl-C.sub.1-C.sub.3alkyl-,
heteroaryl-C.sub.1-C.sub.3alkyl-,
N(R.sup.30)(R.sup.31)-heterocyclyl-C.sub.1-C.sub.3alkyl-, wherein
the aryl, cycloalkyl, heteroaryl and heterocycyl are optionally
substituted with from 1 to 3 substituents from (a); and Y.sup.31 is
selected from the group consisting of a direct bond, --O--,
--N(R.sup.30)--, --C(O)--, --O--C(O)--, --C(O)--O--,
--N(R.sup.30)--C(O)--, --C(O)--N(R.sup.30)--,
--N(R.sup.30)--C(S)--, --C(S)--N(R.sup.30)--,
--N(R.sup.30)--C(O)--N(R.sup.31)--,
--N(R.sup.30)--C(NR.sup.30)--N(R.sup.31)--,
--N(R.sup.30)--C(NR.sup.31)--, --C(NR.sup.31)--N(R.sup.30),
--N(R.sup.30)--C(S)--N(R.sup.31)--, --N(R.sup.30)--C(O)--O--,
--O--C(O)--N(R.sup.31)--, --N(R.sup.30)--C(S)--O--,
--O--C(S)--N(R.sup.31)--, --S(O).sub.0-2--,
--SO.sub.2N(R.sup.31)--, --N(R.sup.31)--SO.sub.2-- and
--N(R.sup.30)--SO.sub.2N(R.sup.31)--.
[0104] As a non-limiting example, substituted phenyls include
2-fluorophenyl, 3,4-dichlorophenyl, 3-chloro-4-fluoro-phenyl,
2-fluoro-3-propylphenyl. As another non-limiting example,
substituted n-octyls include 2,4-dimethyl-5-ethyl-octyl and
3-cyclopentyl-octyl. Included within this definition are methylenes
(--CH.sub.2--) substituted with oxygen to form carbonyl --CO--.
[0105] When there are two optional substituents bonded to adjacent
atoms of a ring structure, such as for example phenyl, thiophenyl,
or pyridinyl, the substituents, together with the atoms to which
they are bonded, optionally form a 5- or 6-membered cycloalkyl or
heterocycle having 1, 2, or 3 annular heteroatoms.
[0106] In a preferred embodiment, hydrocarbyl, alkyl, alkenyl,
alkynyl, heteroalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl,
aromatic polycycle, non-aromatic polycycle, polyheteroaryl,
non-aromatic polyheterocyclic and mixed aryl and non-aryl
polyheterocycle groups are unsubstituted.
[0107] In other preferred embodiments, hydrocarbyl, alkyl, alkenyl,
alkynyl, heteroalkyl, cycloalkyl, heterocyclic, aryl, heteroaryl,
aromatic polycycle, non-aromatic polycycle, polyheteroaryl,
non-aromatic polyheterocyclic and mixed aryl and non-aryl
polyheterocycle groups are substituted with from 1 to 3
independently selected substituents.
[0108] Preferred substituents on alkyl groups include, but are not
limited to, hydroxyl, halogen (e.g., a single halogen substituent
or multiple halo substituents; in the latter case, groups such as
CF.sub.3 or an alkyl group bearing more than one Cl), cyano, nitro,
alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle,
aryl, --OR.sup.u, --SR.sup.u, --S(.dbd.O)R.sup.y,
--S(.dbd.O).sub.2R.sup.y, --P(.dbd.O).sub.2R.sup.y,
--S(.dbd.O).sub.2OR.sup.y, --P(.dbd.O).sub.2OR.sup.y,
--NR.sup.vR.sup.w, --NR.sup.vS(.dbd.O).sub.2R.sup.y,
--NR.sup.vP(.dbd.O).sub.2R.sup.y, --S(.dbd.O).sub.2NR.sup.vR.sup.w,
--P(.dbd.O).sub.2NR.sup.vR.sup.w, --C(.dbd.O)OR.sup.y,
--C(.dbd.O)R.sup.u, --C(.dbd.O)NR.sup.vR.sup.w,
--OC(.dbd.O)R.sup.u, --OC(.dbd.O)NR.sup.vR.sup.w,
--NR.sup.vC(.dbd.O)OR.sup.y, --NR.sup.xxC(.dbd.O)NR.sup.vR.sup.w,
--NR.sup.xxS(.dbd.O).sub.2NR.sup.vR.sup.w,
--NR.sup.xxP(.dbd.O).sub.2NR.sup.vR.sup.w,
--NR.sup.vC(.dbd.O)R.sup.u or --NR.sup.vP(.dbd.O).sub.2R.sup.y,
wherein R.sup.u is hydrogen, alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, heterocycle or aryl; R.sup.v, R.sup.w and
R.sup.xx are independently hydrogen, alkyl, cycloalkyl, heterocycle
or aryl, or said R.sup.v and R.sup.w together with the N to which
they are bonded optionally form a heterocycle; and R.sup.y is
alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle or
aryl. In the aforementioned exemplary substituents, groups such as
alkyl, cycloalkyl, alkenyl, alkynyl, cycloalkenyl, heterocycle and
aryl can themselves be optionally substituted.
[0109] Preferred substituents on alkenyl and alkynyl groups
include, but are not limited to, alkyl or substituted alkyl, as
well as those groups recited as preferred alkyl substituents.
[0110] Preferred substituents on cycloalkyl groups include, but are
not limited to, nitro, cyano, alkyl or substituted alkyl, as well
as those groups recited about as preferred alkyl substituents.
Other preferred substituents include, but are not limited to,
spiro-attached or fused cyclic substituents, preferably
spiro-attached cycloalkyl, spiro-attached cycloalkenyl,
spiro-attached heterocycle (excluding heteroaryl), fused
cycloalkyl, fused cycloalkenyl, fused heterocycle, or fused aryl,
where the aforementioned cycloalkyl, cycloalkenyl, heterocycle and
aryl substituents can themselves be optionally substituted.
[0111] Preferred substituents on cycloalkenyl groups include, but
are not limited to, nitro, cyano, alkyl or substituted alkyl, as
well as those groups recited as preferred alkyl substituents. Other
preferred substituents include, but are not limited to,
spiro-attached or fused cyclic substituents, especially
spiro-attached cycloalkyl, spiro-attached cycloalkenyl,
spiro-attached heterocycle (excluding heteroaryl), fused
cycloalkyl, fused cycloalkenyl, fused heterocycle, or fused aryl,
where the aforementioned cycloalkyl, cycloalkenyl, heterocycle and
aryl substituents can themselves be optionally substituted.
[0112] Preferred substituents on aryl groups include, but are not
limited to, nitro, cycloalkyl or substituted cycloalkyl,
cycloalkenyl or substituted cycloalkenyl, cyano, alkyl or
substituted alkyl, as well as those groups recited above as
preferred alkyl substituents. Other preferred substituents include,
but are not limited to, fused cyclic groups, especially fused
cycloalkyl, fused cycloalkenyl, fused heterocycle, or fused aryl,
where the aforementioned cycloalkyl, cylcoalkenyl, heterocycle and
aryl substituents can themselves be optionally substituted. Still
other preferred substituents on aryl groups (phenyl, as a
non-limiting example) include, but are not limited to, haloalkyl
and those groups recited as preferred alkyl substituents.
[0113] Preferred substituents on heterocylic groups include, but
are not limited to, cycloalkyl, substituted cycloalkyl,
cycloalkenyl, substituted cycloalkenyl, nitro, oxo (i.e., .dbd.O),
cyano, alkyl, substituted alkyl, as well as those groups recited as
preferred alkyl substituents. Other preferred substituents on
heterocyclic groups include, but are not limited to, spiro-attached
or fused cylic substituents at any available point or points of
attachment, more preferably spiro-attached cycloalkyl,
spiro-attached cycloalkenyl, spiro-attached heterocycle (excluding
heteroaryl), fused cycloalkyl, fused cycloakenyl, fused heterocycle
and fused aryl, where the aforementioned cycloalkyl, cycloalkenyl,
heterocycle and aryl substituents can themselves be optionally
substituted.
[0114] In a preferred embodiment, a heterocyclic group is
substituted on carbon, nitrogen and/or sulfur at one or more
positions. Preferred substituents on nitrogen include, but are not
limited to N-oxide, alkyl, aryl, aralkyl, alkylcarbonyl,
alkylsulfonyl, arylcarbonyl, arylsulfonyl, alkoxycarbonyl, or
aralkoxycarbonyl. Preferred substituents on sulfur include, but are
not limited to, oxo and C.sub.1-6alkyl. In certain preferred
embodiments, nitrogen and sulfur heteroatoms may independently be
optionally oxidized and nitrogen heteroatoms may independently be
optionally quaternized.
[0115] Especially preferred substituents on alkyl groups include
halogen and hydroxy.
[0116] Especially preferred substituents on ring groups, such as
aryl, heteroaryl, cycloalkyl and heterocyclyl, include halogen,
alkoxy and alkyl.
[0117] Preferred substituents on aromatic polycycles include, but
are not limited to, oxo, C.sub.1-C.sub.6alkyl, cycloalkylalkyl
(e.g. cyclopropylmethyl), oxyalkyl, halo, nitro, amino, alkylamino,
aminoalkyl, alkyl ketones, nitrile, carboxyalkyl, alkylsulfonyl,
arylsulfonyl, aminosulfonyl and OR.sup.aa, such as alkoxy, wherein
R.sup.aa is selected from the group consisting of H,
C.sub.1-C.sub.6alkyl, C.sub.4-C.sub.9cycloalkyl,
C.sub.4-C.sub.9heterocycloalkyl, aryl, heteroaryl, arylalkyl,
heteroarylalkyl and (CH.sub.2).sub.0-6Z.sup.aR.sup.bb, wherein
Z.sup.a is selected from the group consisting of O, NR.sup.cc, S
and S(O), and R.sup.bb is selected from the group consisting of H,
C.sub.1-C.sub.6alkyl, C.sub.4-C.sub.9cycloalkyl,
C.sub.4-C.sub.9heterocycloalkyl,
C.sub.4-C.sub.9heterocycloalkylalkyl, aryl, mixed aryl and non-aryl
polycycle, heteroaryl, arylalkyl, (e.g. benzyl), and
heteroarylalkyl (e.g. pyridylmethyl); and R.sup.cc is selected from
the group consisting of H, C.sub.1-C.sub.6alkyl,
C.sub.4-C.sub.9cycloalkyl, C.sub.4-C.sub.9heterocycloalkyl, aryl,
heteroaryl, arylalkyl (e.g. benzyl), heteroarylalkyl (e.g.
pyridylmethyl) and amino acyl.
[0118] Preferred substituents on non-aromatic polycycles include,
but are not limited to, oxo, C.sub.3-C.sub.9cycloalkyl, such as
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.
Unless otherwise noted, non-aromatic polycycle substituents include
both unsubstituted cycloalkyl groups and cycloalkyl groups that are
substituted by one or more suitable substituents, including but not
limited to, C.sub.1-C.sub.6alkyl, oxo, halo, hydroxy, aminoalkyl,
oxyalkyl, alkylamino and OR.sup.aa, such as alkoxy. Preferred
substituents for such cycloalkyl groups include halo, hydroxy,
alkoxy, oxyalkyl, alkylamino and aminoalkyl.
[0119] Preferred substituents on carbon atoms of polyheteroaryl
groups include but are not limited to, straight and branched
optionally substituted C.sub.1-C.sub.6alkyl, unsaturation (i.e.,
there are one or more double or triple C--C bonds), acyl, oxo,
cycloalkyl, halo, oxyalkyl, alkylamino, aminoalkyl, acylamino,
OR.sup.aa (for example alkoxy), and a substituent of the formula
--O--(CH.sub.2CH.dbd.CH(CH.sub.3)(CH.sub.2)).sub.1-3H. Examples of
suitable straight and branched C.sub.1-C.sub.6alkyl substituents
include but are not limited to methyl, ethyl, n-propyl, 2-propyl,
n-butyl, sec-butyl, t-butyl and the like. Preferred substituents
include halo, hydroxy, alkoxy, oxyalkyl, alkylamino and aminoalkyl.
Preferably substitutions on nitrogen atoms include, for example by
N-oxide or R.sup.cc. Preferred substituents on nitrogen atoms
include H, C.sub.1-C.sub.4alkyl, acyl, aminoacyl and sulfonyl.
Preferably sulfur atoms are unsubstituted. Preferred substituents
on sulfur atoms include but are not limited to oxo and lower
alkyl.
[0120] Preferred substituents on carbon atoms of non-aromatic
polyheterocyclic groups include but are not limited to straight and
branched optionally substituted C.sub.1-C.sub.6alkyl, unsaturation
(i.e., there are one or more double or triple C--C bonds), acyl,
oxo, cycloalkyl, halo, oxyalkyl, alkylamino, aminoalkyl, acylamino
and OR.sup.aa, for example alkoxy. Examples of suitable straight
and branched C.sub.1-C.sub.6alkyl substituents include but are not
limited to methyl, ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl,
t-butyl and the like. Preferred substituents include halo, hydroxy,
alkoxy, oxyalkyl, alkylamino and aminoalkyl. Preferably
substitutions on nitrogen atoms include, for example, N-oxide or
R.sup.cc. Preferred N substituents include H, C.sub.1-C.sub.4
alkyl, acyl, aminoacyl and sulfonyl. Preferably, sulfur atoms are
unsubstituted. Preferred S substituents include oxo and lower
alkyl.
[0121] Preferred substituents on mixed aryl and non-aryl
polyheterocycle groups include, but are not limited to, nitro or as
described above for non-aromatic polycycle groups. Preferred
substituents on carbon atoms include, but are not limited to,
--N--OH, .dbd.N--OH, optionally substituted alkyl, unsaturation
(i.e., there are one or more double or triple C--C bonds), oxo,
acyl, cycloalkyl, halo, oxyalkyl, alkylamino, aminoalkyl, acylamino
and OR.sup.aa, for example alkoxy. Preferably substitutions on
nitrogen atoms include, for example, N-oxide or R.sup.cc. Preferred
N substituents include H, C.sub.1-4alkyl, acyl aminoacyl and
sulfonyl. Preferably sulfur atoms are unsubstituted. Preferred S
substituents include oxo and lower alkyl.
[0122] A "halohydrocarbyl" is a hydrocarbyl moiety in which from
one to all hydrogens have been replaced with one or more halo.
[0123] The term "halogen" or "halo" is intended to mean chlorine,
bromine, fluorine, or iodine. As herein employed, the term "acyl"
refers to an alkylcarbonyl or arylcarbonyl substituent. The term
"acylamino" refers to an amide group attached at the nitrogen atom
(i.e., R--CO--NH--). The term "carbamoyl" refers to an amide group
attached at the carbonyl carbon atom (i.e., NH.sub.2--CO--). The
nitrogen atom of an acylamino or carbamoyl substituent is
additionally optionally substituted. The term "sulfonamido" refers
to a sulfonamide substituent attached by either the sulfur or the
nitrogen atom. The term "amino" is meant to include NH.sub.2,
alkylamino, arylamino, and cyclic amino groups. The term "ureido"
as employed herein refers to a substituted or unsubstituted urea
moiety.
[0124] The term "radical" is intended to mean a chemical moiety
comprising one or more unpaired electrons.
[0125] Where optional substituents are chosen from "one or more"
groups it is to be understood that this definition includes all
substituents being chosen from one of the specified groups or the
substituents being chosen from two or more of the specified
groups.
[0126] In addition, substituents on cyclic moieties (i.e.,
cycloalkyl, heterocyclyl, aryl, heteroaryl) include 5-6 membered
mono- and 9-14 membered bi-cyclic moieties fused to the parent
cyclic moiety to form a bi- or tri-cyclic fused ring system.
Substituents on cyclic moieties also include 5-6 membered mono- and
9-14 membered bi-cyclic moieties attached to the parent cyclic
moiety by a covalent bond to form a bi- or tri-cyclic bi-ring
system. For example, an optionally substituted phenyl includes, but
is not limited to, the following:
##STR00004##
[0127] An "unsubstituted" moiety (e.g., unsubstituted cycloalkyl,
unsubstituted heteroaryl, etc.) means that moiety as defined above
that does not have an optional substituent. Thus, for example,
"unsubstituted aryl" does not include phenyl substituted with a
halo.
[0128] As used herein, "an amino protecting group" refers to any
functional group commonly used to protect an .alpha.-amino group.
Suitable amino protecting groups include, but are not limited to,
t-butyloxycarbonyl, isoamyloxycarbonyl, o-nitrophenylsulfenyl,
fluoroenylmethyloxycarbonyl, o-nitropyridinylsulfenyl and
biphenylproploxycarbonyl.
[0129] An "amino acid residue" refers to any residue of a natural
or unnatural amino acid, non-limiting examples of which are
residues of alanine, arginine, asparagine, aspartic acid, cysteine,
homocysteine, glutamine, glutamic acid, isoleucine, norleucine,
glycine, phenylglycine, leucine, histidine, methionine, lysine,
phenylalanine, homophenylalanine, ornithine, praline, serine,
homoserine, valine, norvaline, threonine, tryptophane, tyrosine and
the like. With the exception of glycine, all amino acids may be in
the D-, L- or D,L-form.
[0130] The term "radical" is intended to mean a chemical moiety
comprising one or more unpaired electrons.
[0131] Some compounds of the invention may have one or more chiral
centers and/or geometric isomeric centers (E- and Z-isomers), and
it is to be understood that the invention encompasses all such
optical, diastereoisomers and geometric isomers. The invention also
comprises all tautomeric forms of the compounds disclosed
herein.
[0132] All of the compounds in this application were named using
Chemdraw Ultra version 9 or 10, which are available through
Cambridgesoft.co, 100 Cambridge Park Drive, Cambridge, Mass.
02140.
Compounds
[0133] In a first aspect, the invention provides prodrugs of
inhibitors of histone deacetylase, the prodrugs having the formula
(1):
Cy-L.sup.1-Ar-Y.sup.1--C(O)--N(R.sup.x)--Z (1)
[0134] and pharmaceutically acceptable salts thereof, wherein
[0135] Cy is --H, cycloalkyl, aryl, heteroaryl, or heterocyclyl,
any of which may be optionally substituted;
[0136] L.sup.1 is --(CH.sub.2).sub.m--W--, where m is 0, 1, 2, 3,
or 4, and W is selected from the group consisting of --C(O)NH--,
--S(O).sub.2NH--, --NHC(O)--, --NHS(O).sub.2--, and
--NH--C(O)--NH--;
[0137] Ar is arylene, wherein said arylene optionally may be
additionally substituted and optionally may be fused to an aryl or
heteroaryl ring, or to a saturated or partially unsaturated
cycloalkyl or heterocyclic ring, any of which may be optionally
substituted;
[0138] Y.sup.1 is a chemical bond or a straight- or branched-chain
saturated alkylene, wherein said alkylene may be optionally
substituted;
[0139] R.sup.x is H or --OH; [0140] Z is --R.sup.20, --O--R.sup.20,
--R.sup.21, or
##STR00005##
[0140] wherein --R.sup.20 is selected from the group consisting of
--C(O)--R.sup.10, --C(O)O--R.sup.10, --R.sup.11,
--CH(R.sup.12)--O--C(O)--R.sup.10,
--C(O)--C[(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13),
--S(O.sub.2)R.sup.10--C(O)--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.-
10,
--C(O)--(CH.sub.2).sub.1-4--C(OH)(COOR.sup.10)--(CH.sub.2).sub.1-4--CO-
OR.sup.10, --C(O)-[C(R.sup.14)(R.sup.14)].sub.14--P(O)(OH)(OH),
--C(O)--(CH.sub.2).sub.1-4--N(R.sup.14)--C[.dbd.N(R.sup.10')]-N(R.sup.10'-
)(R.sup.10'),
--C(O)--(CH.sub.2)--CH(OH)--(CH.sub.2)--N(CH.sub.3)(CH.sub.3),
--C(O)--CH(NH.sub.2)--(CH.sub.2).sub.1-6--COOH (preferably
--C(O)--CH(NH.sub.2)--(CH.sub.2)--COOH),
--C(O)--O--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.10 and
--C(O)--(CH.sub.2).sub.n--C(O)OR.sup.10, provided that the N to
which Z is bound is not directly bonded to two O atoms and further
provided that (a) when Z is --R.sup.20 then R.sup.x is --OH, and
(b) when Z is --OR.sup.20 then R.sup.x is --H; or
[0141] R.sup.x is absent and R.sup.20 forms an optionally
substituted heterocyclic ring with the N to which it is
attached;
[0142] n is 0, 1, 2, 3, or 4, preferably 1, 2, 3, or 4;
[0143] each R.sup.10 is independently selected from the group
consisting of hydrogen, optionally substituted C.sub.1-C.sub.20
alkyl, optionally substituted C.sub.2-C.sub.20 alkenyl, optionally
substituted C.sub.2-C.sub.20 alkynyl, optionally substituted
C.sub.1-C.sub.20 alkoxycarbonyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, optionally substituted heteroaryl, optionally substituted
cycloalkylalkyl, optionally substituted heterocycloalkylalkyl,
optionally substituted arylalkyl, optionally substituted
heteroarylalkyl, optionally substituted cycloalkylalkenyl,
optionally substituted heterocycloalkylalkenyl, optionally
substituted arylalkenyl, optionally substituted heteroarylalkenyl,
optionally substituted cycloalkylalkynyl, optionally substituted
heterocycloalkylalkynyl, optionally substituted arylalkynyl,
optionally substituted heteroarylalkynyl, optionally substituted
alkylcycloalkyl, optionally substituted alkylheterocycloalkyl,
optionally substituted alkylaryl, optionally substituted
alkylheteroaryl, optionally substituted alkenylcycloalkyl,
optionally substituted alkenylheterocycloalkyl, optionally
substituted alkenylaryl, optionally substituted alkenylheteroaryl,
optionally substituted alkynylcycloalkyl, optionally substituted
alkynylheterocycloalkyl, optionally substituted alkynylary,
optionally substituted alkynylheteroaryl, a sugar residue, and an
amino acid residue (preferably bonded through the carboxy terminus
of the amino acid);
[0144] each R.sup.10' is independently hydrogen or C.sub.1-6alkyl,
or
[0145] R.sup.10 and R.sup.10' together with the carbon atom to
which they are attached form an optionally substituted
spirocycloalkyl;
[0146] R.sup.21 is a sugar or -amino acid-R.sup.13, wherein
R.sup.13 is covalently bound to the N-terminus;
[0147] R.sup.11 is selected from the group consisting of hydrogen,
optionally substituted heterocycloalkyl, optionally substituted
aryl, and optionally substituted heteroaryl;
[0148] R.sup.12 is selected from hydrogen or alkyl; and
[0149] R.sup.13 is selected from the group consisting of hydrogen,
--C(O)--CH[N(R.sup.10')(R.sup.10')]--C.sub.1-C.sub.6alkyl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-N(R.sup.10')(R.s-
up.10'),
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-aryl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-heteroaryl,
--C(O)-aryl, --C(O)-heteroaryl, an amino protecting group, and
R.sup.10; and
[0150] each R.sup.14 is independently selected from the group
consisting of H, C.sub.1-C.sub.6alkyl and cycloalkyl, or two
R.sup.14, together with the atom to which they are attached, form a
cycloalkyl.
[0151] In certain preferred embodiments, Cy is C.sub.6-C.sub.14
aryl, more preferably C.sub.6-C.sub.10 aryl, and most preferably
phenyl or naphthyl, any of which may be optionally substituted. In
certain other preferred embodiments, Cy is heteroaryl. In some
preferred embodiments, the heteroaryl group is selected from the
group consisting of thienyl, benzothienyl, furyl, benzofuryl,
quinolyl, isoquinolyl, and thiazolyl, any of which may be
optionally substituted. In certain particularly preferred
embodiments, Cy is selected from the group consisting of phenyl,
naphthyl, thienyl, benzothienyl, and quinolyl, any of which may be
optionally substituted. In certain other preferred embodiments, Cy
is phenyl, pyridine or indole, more preferably phenyl or indole. In
certain preferred embodiments, Cy is substituted with one or more
substituents selected from the group consisting of trihaloalkyl
(preferably trifluoroalkyl), halogen, CN, amidine, sulfone,
alkylsulfone, imidate and alkylimidate. In certain preferred
embodiments, Cy is phenyl substituted with one or more substituents
selected from the group consisting of trihaloalkyl (preferably
trifluoroalkyl), halogen, CN, amidine, sulfone, alkylsulfone,
imidate and alkylimidate, preferably selected from the group
consisting of trihaloalkyl (preferably trifluoroalkyl) and
halogen.
[0152] L.sup.1 is --(CH.sub.2).sub.m--W--, where m is 0, 1, 2, 3,
or 4, and W is selected from the group consisting of --C(O)NH--,
--S(O).sub.2NH--, --NHC(O)--, --NHS(O).sub.2--, and
--NH--C(O)--NH--. Preferably, m is 0, 1, or 2, more preferably 0 or
1.
[0153] Preferably, Ar is C.sub.6-C.sub.14 arylene, more preferably
C.sub.6-C.sub.10 arylene, any of which may be additionally
substituted. In certain preferred embodiments, Ar is phenylene,
preferably 4-phenylene. In some preferred embodiments, the
phenylene is fused to an aryl or heteroaryl ring, or to a saturated
or partially unsaturated cycloalkyl or heterocyclic ring, any of
which groups also may be optionally substituted.
[0154] Y.sup.1 is a chemical bond or is a straight- or
branched-chain alkylene, which may be optionally substituted. In
some preferred embodiments, Y.sup.1 is a chemical bond, and the
group --C(O)NH--Z is directly attached to Ar. In some other
preferred embodiments, Y.sup.1 is alkylene, preferably saturated
alkylene. Preferably, the saturated alkylene is C.sub.1-C.sub.8
alkylene, more preferably C.sub.1-C.sub.6 alkylene, still more
preferably C.sub.1-C.sub.3 alkylene, and yet still more preferably
C.sub.1-C.sub.2 alkylene, any of which may be optionally
substituted. In some particularly preferred embodiments, Y.sup.1 is
methylene.
[0155] Substituted alkyl, aryl, heterocyclyl, and heteroaryl groups
have one or more, preferably between one and about three, more
preferably one or two substituents, which are preferably selected
from the group consisting of C.sub.1-C.sub.6 alkyl, preferably
C.sub.1-C.sub.4 alkyl; halo, preferably Cl, Br, or F; haloalkyl,
preferably (halo).sub.1-5(C.sub.1-C.sub.6)alkyl, more preferably
(halo).sub.1-5(C.sub.1-C.sub.3)alkyl, and most preferably CF.sub.3;
C.sub.1-C.sub.6 alkoxy, preferably methoxy, ethoxy, or benzyloxy;
C.sub.6-C.sub.10 aryloxy, preferably phenoxy; C.sub.1-C.sub.6
alkoxycarbonyl, preferably C.sub.1-C.sub.3 alkoxycarbonyl, most
preferably carbomethoxy or carboethoxy; C.sub.6-C.sub.10 aryl,
preferably phenyl; (C.sub.6-C.sub.10)ar(C.sub.1-C.sub.6)alkyl,
preferably (C.sub.6-C.sub.10)ar(C.sub.1-C.sub.3)alkyl, more
preferably benzyl, naphthylmethyl or phenethyl;
hydroxy(C.sub.1-C.sub.6)alkyl, preferably
hydroxy(C.sub.1-C.sub.3)alkyl, more preferably hydroxymethyl;
amino(C.sub.1-C.sub.6)alkyl, preferably
amino(C.sub.1-C.sub.3)alkyl, more preferably aminomethyl;
(C.sub.1-C.sub.6)alkylamino, preferably methylamino, ethylamino, or
propylamino; di-(C.sub.1-C.sub.6)alkylamino, preferably
dimethylamino or diethylamino; (C.sub.1-C.sub.6)alkylcarbamoyl,
preferably methylcarbamoyl, dimethylcarbamoyl, or benzylcarbamoyl;
(C.sub.6-C.sub.10)arylcarbamoyl, preferably phenylcarbamoyl;
(C.sub.1-C.sub.6)alkaneacylamino, preferably acetylamino;
(C.sub.6-C.sub.10)areneacylamino, preferably benzoylamino;
(C.sub.1-C.sub.6)alkanesulfonyl, preferably methanesulfonyl;
(C.sub.1-C.sub.6)alkanesulfonamido, preferably methanesulfonamido;
(C.sub.6-C.sub.10)arenesulfonyl, preferably benzenesulfonyl or
toluenesulfonyl; (C.sub.6-C.sub.10)arenesulfonamido, preferably
benzenesulfonyl or toluenesulfonyl;
(C.sub.6-C.sub.10)ar(C.sub.1-C.sub.6)alkylsulfonamido, preferably
benzylsulfonamido; C.sub.1-C.sub.6 alkylcarbonyl, preferably
C.sub.1-C.sub.3 alkylcarbonyl, more preferably acetyl;
(C.sub.1-C.sub.6)acyloxy, preferably acetoxy; cyano; amino;
carboxy; hydroxy; ureido; and nitro. One or more carbon atoms of an
alkyl, cycloalkyl, or heterocyclyl group may also be optionally
substituted with an oxo group.
[0156] In some particularly preferred embodiments, Cy is a phenyl,
naphthyl, thienyl, benzothienyl, or quinolyl moiety which is
unsubstituted or is substituted by one or two substituents
independently selected from the group consisting of C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 haloalkyl, C.sub.6-C.sub.10 aryl,
(C.sub.6-C.sub.10)ar(C.sub.1-C.sub.6)alkyl, halo, nitro, hydroxy,
C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkoxycarbonyl, carboxy,
and amino
[0157] In some preferred embodiments, Z is -O--C(O)--R.sup.10,
--O--C(O)-[C(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13) or
--OR.sup.11.
[0158] In some preferred embodiments, the amino acid is an L-amino
acid.
[0159] In certain preferred embodiments, the sugar residue is a
saccharide selected from the group consisting of glucose,
galactose, mannose, gulose, idose, talose, allose, altrose,
fructose, rhamnose, ribose and xylose.
[0160] In a second embodiment, the invention provides prodrugs of
inhibitors of histone deacetylase, the prodrugs represented by
formula (2):
Cy-L.sup.2-Ar-Y.sup.2--C(O)N(R.sup.x)--Z (2)
[0161] and pharmaceutically acceptable salts thereof, wherein
[0162] Cy is H, cycloalkyl, aryl, heteroaryl, or heterocyclyl, any
of which may be optionally substituted, provided that Cy is not a
(spirocycloalkyl)heterocyclyl;
[0163] L.sup.2 is C.sub.1-C.sub.6 saturated alkylene or
C.sub.2-C.sub.6 alkenylene, wherein the alkylene or alkenylene
optionally may be substituted, and wherein one or two of the carbon
atoms of the alkylene is optionally replaced by a heteroatomic
moiety independently selected from the group consisting of 0; NR',
R' being alkyl, acyl, or hydrogen; S; S(O); or S(O).sub.2;
[0164] Ar is arylene, wherein said arylene optionally may be
additionally substituted and optionally may be fused to an aryl or
heteroaryl ring, or to a saturated or partially unsaturated
cycloalkyl or heterocyclic ring, any of which may be optionally
substituted; and
[0165] Y.sup.2 is a chemical bond or a straight- or branched-chain
saturated alkylene, which may be optionally substituted, provided
that the alkylene is not substituted with a substituent of the
formula --C(O)R wherein R comprises an .alpha.-amino acyl
moiety;
[0166] R.sup.x is H or --OH;
[0167] Z is --R.sup.20, --O--R.sup.20, --R.sup.21, or
##STR00006##
wherein --R.sup.20 is selected from the group consisting of
--C(O)--R.sup.10, --C(O)O--R.sup.10, --R.sup.11,
--CH(R.sup.12)--O--C(O)--R.sup.10,
--C(O)--C[(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13),
--S(O.sub.2)R.sup.10--C(O)--(CH.sub.2).sub.1-4--C(OH)(COOR.sup.10)--(CH.s-
ub.2).sub.14--COOR.sup.10,
--C(O)-[C(R.sup.14)(R.sup.14)].sub.1-4--P(O)(OH)(OH),
--C(O)--(CH.sub.2).sub.1-4--N(R.sup.14)--C[.dbd.N(R.sup.10')]-N(R.sup.10'-
)(R.sup.10'),
--C(O)--(CH.sub.2)--CH(OH)--(CH.sub.2)--N(CH.sub.3)(CH.sub.3),
--C(O)--CH(NH.sub.2)--(CH.sub.2).sub.1-6--COOH (preferably
--C(O)--CH(NH.sub.2)--(CH.sub.2)--COOH),
--C(O)--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.10,
--C(O)--O--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.10 and
--C(O)--(CH.sub.2).sub.n--C(O)OR.sup.10, provided that the N to
which Z is bound is not directly bonded to two O atoms; and further
provided that (a) when Z is --R.sup.20 then R.sup.x is --OH, and
(b) when Z is --OR.sup.20 then R.sup.x is --H; or
[0168] R.sup.x is absent and R.sup.20 forms an optionally
substituted heterocyclic ring with the N to which it is
attached;
[0169] n is 0, 1, 2, 3, or 4, preferably 1, 2, 3, or 4;
[0170] each R.sup.10 is independently selected from the group
consisting of hydrogen, optionally substituted C.sub.1-C.sub.20
alkyl, optionally substituted C.sub.2-C.sub.20 alkenyl, optionally
substituted C.sub.2-C.sub.20 alkynyl, optionally substituted
C.sub.1-C.sub.20 alkoxycarbonyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, optionally substituted heteroaryl, optionally substituted
cycloalkylalkyl, optionally substituted heterocycloalkylalkyl,
optionally substituted arylalkyl, optionally substituted
heteroarylalkyl, optionally substituted cycloalkylalkenyl,
optionally substituted heterocycloalkylalkenyl, optionally
substituted arylalkenyl, optionally substituted heteroarylalkenyl,
optionally substituted cycloalkylalkynyl, optionally substituted
heterocycloalkylalkynyl, optionally substituted arylalkynyl,
optionally substituted heteroarylalkynyl, optionally substituted
alkylcycloalkyl, optionally substituted alkylheterocycloalkyl,
optionally substituted alkylaryl, optionally substituted
alkylheteroaryl, optionally substituted alkenylcycloalkyl,
optionally substituted alkenylheterocycloalkyl, optionally
substituted alkenylaryl, optionally substituted alkenylheteroaryl,
optionally substituted alkynylcycloalkyl, optionally substituted
alkynylheterocycloalkyl, optionally substituted alkynylary,
optionally substituted alkynylheteroaryl, a sugar residue, and an
amino acid residue (preferably bonded through the carboxy terminus
of the amino acid);
[0171] each R.sup.10' is independently hydrogen or C.sub.1-6alkyl,
or
[0172] R.sup.10 and R.sup.10' together with the carbon atom to
which they are attached form an optionally substituted
spirocycloalkyl;
[0173] R.sup.21 is a sugar or -amino acid-R.sup.13, wherein
R.sup.13 is covalently bound to the N-terminus;
[0174] R.sup.11 is selected from the group consisting of hydrogen,
optionally substituted heterocycloalkyl, optionally substituted
aryl, and optionally substituted heteroaryl;
[0175] R.sup.12 is selected from hydrogen or alkyl; and
[0176] R.sup.13 is selected from the group consisting of hydrogen,
--C(O)--CH[N(R.sup.10')(R.sup.10')]C.sub.1-C.sub.6alkyl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-N(R.sup.10')(R.s-
up.10')C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-aryl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]--C.sub.1-C.sub.6alkyl-heteroaryl,
--C(O)-aryl, --C(O)-heteroaryl, an amino protecting group, and
R.sup.10; and
[0177] each R.sup.14 is independently selected from the group
consisting of H, C.sub.1-C.sub.6alkyl and cycloalkyl, or two
R.sup.14, together with the atom to which they are attached, form a
cycloalkyl.
[0178] Preferred substituents Cy, Ar, and Z according to this
aspect of the invention are as defined above for the first
embodiment. Preferred substituents of Y.sup.2 are as defined above
for Y.sup.1. In some preferred embodiments, L.sup.2 is saturated
C.sub.1-C.sub.8 alkylene, more preferably C.sub.1-C.sub.6 alkylene,
still more preferably C.sub.1-C.sub.4 alkylene, any of which groups
may be optionally substituted. In some other preferred embodiments,
L.sup.2 is C.sub.2-C.sub.8 alkenylene, more preferably
C.sub.2-C.sub.6 alkenylene, and still more preferably
C.sub.2-C.sub.4 alkenylene, any of which groups may be optionally
substituted. The alkylene or alkenylene group may be substituted at
one or more carbon positions with a substituent preferably selected
from the list of preferred substituents recited above. More
preferably, L.sup.2 is substituted at one or two positions with a
substituent independently selected from the group consisting of
C.sub.1-C.sub.6 alkyl, C.sub.6-C.sub.10 aryl, amino, oxo, hydroxy,
C.sub.1-C.sub.4 alkoxy, and C.sub.6-C.sub.10 aryloxy. In some
particularly preferred embodiments, the alkylene or alkenylene
group is substituted with one or two oxo or hydroxy groups.
[0179] In some preferred embodiments, L.sup.1 is C.sub.1-C.sub.6
saturated alkylene, wherein on of the carbon atoms of the saturated
alkylene is replaced by a heteroatom moiety selected from the group
consisting of O; NR', R' being alkyl, acyl, or hydrogen; S; S(O);
or S(O).sub.2. Preferably, the carbon atom adjacent to Cy is
replaced by a heteroatom moiety. In some particularly preferred
embodiments, L.sup.1 is selected from the group consisting of
--S--(CH.sub.2).sub.2--; --S(O)--(CH.sub.2).sub.2--,
--S(O).sub.2--(CH.sub.2).sub.2--, --S--(CH.sub.2).sub.3--,
--S(O)--(CH.sub.2).sub.3--, and
--S(O).sub.2--(CH.sub.2).sub.3--.
[0180] In some preferred embodiments, Z is --O--C(O)--R.sup.10,
--O--C(O)-[C(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13) or
--OR.sup.11. Even more preferred embodiments of compound (2)
are:
##STR00007##
[0181] In a third embodiment, the invention provides prodrugs of
inhibitors of histone deacetylase, the prodrugs represented by
formula (3):
Cy-L.sup.3-Ar-Y.sup.3--C(O)N(R.sup.x)--Z (3)
[0182] and pharmaceutically acceptable salts thereof, wherein
[0183] Cy is --H, cycloalkyl, aryl, heteroaryl, or heterocyclyl,
any of which may be optionally substituted, provided that Cy is not
a (spirocycloalkyl)heterocyclyl;
[0184] L.sup.3 is selected from the group consisting of
[0185] (a) --(CH.sub.2).sub.m--W--, where m is 0, 1, 2, 3, or 4,
and W is selected from the group consisting of --C(O)NH--,
--S(O).sub.2NH--, --NHC(O)--, --NHS(O).sub.2--, and
--NH--C(O)--NH--; and
[0186] (b) C.sub.1-C.sub.6 alkylene or C.sub.2-C.sub.6 alkenylene,
wherein the alkylene or alkenylene optionally may be substituted,
and wherein one of the carbon atoms of the alkylene optionally may
be replaced by O; NR', R' being alkyl, acyl, or hydrogen; S; S(O);
or S(O).sub.2;
[0187] Ar is arylene, wherein said arylene optionally may be
additionally substituted and optionally may be fused to an aryl or
heteroaryl ring, or to a saturated or partially unsaturated
cycloalkyl or heterocyclic ring, any of which may be optionally
substituted; and
[0188] Y.sup.3 is C.sub.2 alkenylene or C.sub.2 alkynylene, wherein
one or both carbon atoms of the alkenylene optionally may be
substituted with alkyl, aryl, alkaryl, or aralkyl;
[0189] R.sup.x is H or --OH;
[0190] Z is --R.sup.20, --O--R.sup.20, --R.sup.21, or
##STR00008##
wherein --R.sup.20 is selected from the group consisting of
--C(O)--R.sup.10, --C(O)O--R.sup.10, --R.sup.11,
--CH(R.sup.12)--O--C(O)--R.sup.10,
--C(O)--(CH.sub.2).sub.1-4--C(OH)(COOR.sup.10)--(CH.sub.2).sub.1-4--COOR.-
sup.10, --C(O)-[C(R.sup.14)(R.sup.14)].sub.1-4--P(O)(OH)(OH),
--C(O)--(CH.sub.2).sub.1-4--N(R.sup.14)--C[.dbd.N(R.sup.10')]-N(R.sup.10'-
)(R.sup.10')--C(O)--(CH.sub.2)--CH(OH)--(CH.sub.2)--N(CH.sub.3)(CH.sub.3),
--C(O)--CH(NH.sub.2)--(CH.sub.2).sub.1-6--COOH (preferably
--C(O)--CH(NH.sub.2)--(CH.sub.2)--COOH),
--C(O)--C[(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13),
--S(O.sub.2)R.sup.10, --P(O)(OR.sup.10)(OR.sup.10),
--C(O)--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.10,
--C(O)--O--(CH.sub.2).sub.n--CH(OH)--CH.sub.2--O--R.sup.10 and
--C(O)--(CH.sub.2).sub.n--C(O)OR.sup.10, provided that the N to
which Z is bound is not directly bonded to two O atoms; and further
provided that (a) when Z is --R.sup.20 then R.sup.x is --OH, and
(b) when Z is --OR.sup.20 then R.sup.x is --H; or
[0191] R.sup.x is absent and R.sup.20 forms an optionally
substituted heterocyclic ring with the N to which it is
attached;
[0192] n is 0, 1, 2, 3, or 4, preferably 1, 2, 3, or 4;
[0193] each R.sup.10 is independently selected from the group
consisting of hydrogen, optionally substituted C.sub.1-C.sub.20
alkyl, optionally substituted C.sub.2-C.sub.20 alkenyl, optionally
substituted C.sub.2-C.sub.20 alkynyl, optionally substituted
C.sub.1-C.sub.20 alkoxycarbonyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, optionally substituted heteroaryl, optionally substituted
cycloalkylalkyl, optionally substituted heterocycloalkylalkyl,
optionally substituted arylalkyl, optionally substituted
heteroarylalkyl, optionally substituted cycloalkylalkenyl,
optionally substituted heterocycloalkylalkenyl, optionally
substituted arylalkenyl, optionally substituted heteroarylalkenyl,
optionally substituted cycloalkylalkynyl, optionally substituted
heterocycloalkylalkynyl, optionally substituted arylalkynl,
optionally substituted heteroarylalkynyl, optionally substituted
alkylcycloalkyl, optionally substituted alkylheterocycloalkyl,
optionally substituted alkylaryl, optionally substituted
alkylheteroaryl, optionally substituted alkenylcycloalkyl,
optionally substituted alkenylheterocycloalkyl, optionally
substituted alkenylaryl, optionally substituted alkenylheteroaryl,
optionally substituted alkynylcycloalkyl, optionally substituted
alkynylheterocycloalkyl, optionally substituted alkynylary,
optionally substituted alkynylheteroaryl, a sugar residue, and an
amino acid residue (preferably bonded through the carboxy terminus
of the amino acid);
[0194] each R.sup.10' is independently hydrogen or C.sub.1-6alkyl,
or
[0195] R.sup.10 and R.sup.10' together with the carbon atom to
which they are attached form an optionally substituted
spirocycloalkyl;
[0196] R.sup.21 is a sugar or -amino acid-R.sup.13, wherein
R.sup.13 is covalently bound to the N-terminus;
[0197] R.sup.11 is selected from the group consisting of hydrogen,
optionally substituted heterocycloalkyl, optionally substituted
aryl, and optionally substituted heteroaryl;
[0198] R.sup.12 is selected from hydrogen or alkyl; and
[0199] R.sup.13 is selected from the group consisting of hydrogen,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-N(R.sup.10)(R.su-
p.10'),
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-aryl,
--C(O)--CH[N(R.sup.10')(R.sup.10')]-C.sub.1-C.sub.6alkyl-heteroaryl,
--C(O)-aryl, --C(O)-heteroaryl, an amino protecting group, and
R.sup.10; and
[0200] each R.sup.14 is independently selected from the group
consisting of H, C.sub.1-C.sub.6alkyl and cycloalkyl, or two
R.sup.14, together with the atom to which they are attached, form a
cycloalkyl.
[0201] Preferred substituents Cy, Ar, and Z according to this
aspect of the invention are as defined above for the first
embodiment. Preferred substituents L.sup.3 are as defined above for
L.sup.1 or L.sup.2.
[0202] Preferably, Y.sup.3 is C.sub.2 alkenylene or C.sub.2
alkynylene, wherein one or both carbon atoms of the alkenylene
optionally may be substituted with C.sub.1-C.sub.6 alkyl,
C.sub.6-C.sub.10 aryl, (C.sub.1-C.sub.6)alk(C.sub.6-C.sub.10)aryl,
or (C.sub.6-C.sub.10)ar(C.sub.1-C.sub.6)alkyl. More preferably,
Y.sup.3 is C.sub.2 alkenylene or C.sub.2 alkynylene, wherein one or
both carbon atoms of the alkenylene optionally may be substituted
with C.sub.1-C.sub.4 alkyl, C.sub.6-C.sub.10 aryl,
(C.sub.1-C.sub.4)alk(C.sub.6-C.sub.10)aryl, or
(C.sub.6-C.sub.10)ar(C.sub.1-C.sub.4)alkyl. Still more preferably,
Y.sup.3 is selected from the group consisting of --C.ident.C--,
--CH.dbd.CH--, --C(CH.sub.3).dbd.CH--, and
--CH.dbd.C(CH.sub.3)--.
[0203] In a preferred embodiment of the compounds of formulae (1),
(2), and (3), Z is selected from the group consisting of
--O--C(O)--(CH.sub.2).sub.1-4--C(OH)(COOR.sup.10)--(CH.sub.2).sub.14--COO-
R.sup.10, --O--C(O)-[C(R.sup.14)(R.sup.14)].sub.1-4--P(O)(OH)(OH),
--O--C(O)--(CH.sub.2).sub.14--N(R.sup.14)--C[.dbd.N(R.sup.10')]-N(R.sup.1-
0')(R.sup.10'),
--O--C(O)--(CH.sub.2)--CH(OH)--(CH.sub.2)--N(CH.sub.3)(CH.sub.3),
--O--C(O)--CH(NH.sub.2)--(CH.sub.2).sub.1-6--COOH, preferably
--O--C(O)--CH(NH.sub.2)--(CH.sub.2)--COOH.
[0204] In a preferred embodiment of the compounds of formulae (1),
(2), and (3), Z is selected from the group consisting of
--O--C(O)--(CH.sub.2)--C(OH)(COOH)--(CH.sub.2)--COOH,
--O--C(O)--CH.sub.2--P(O)(OH)(OH),
--O--C(O)--(CH.sub.2)--N(CH.sub.3)--C(.dbd.NH)--NH.sub.2,
--O--C(O)--(CH.sub.2)--CH(OH)--(CH.sub.2)--N(CH.sub.3)(CH.sub.3),
--O--C(O)--CH(NH.sub.2)--(CH.sub.2)--COOH.
[0205] In some preferred embodiments, Z is --O--C(O)--R.sup.10,
--O--C--(O)-[C(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13) or
--OR.sup.11.
[0206] In some preferred embodiments of the prodrugs of inhibitors
of histone deacetylase, of formulae (1), (2), and (3), Z is
--O--R.sup.20 wherein R.sup.20 is
--C(O)--CR.sup.10R.sup.10'--NH(R.sup.13), R.sup.13 and R.sup.10'
are H, and R.sup.10 is C.sub.1-C.sub.6-alkyl or an amino acid side
chain, or R.sup.10 and R.sup.10' together with the carbon to which
they are linked form C.sub.3-C.sub.6 cycloalkyl.
[0207] Naturally-occurring or non-naturally occurring amino acids
are used to prepare the prodrugs of the invention. In particular,
standard amino acids suitable as a prodrug moiety include valine,
leucine, isoleucine, methionine, phenylalanine, asparagine,
glutamic acid, glutamine, histidine, lysine, arginine, aspartic
acid, glycine, alanine, serine, threonine, tyrosine, tryptophan,
cysteine and proline. Particularly preferred are L-amino acids.
Optionally an included amino acid is an .alpha.-, .beta.-, or
.gamma.-amino acid. Also, naturally-occurring, non-standard amino
acids can be utilized in the compositions and methods of the
invention. For example, in addition to the standard naturally
occurring amino acids commonly found in proteins, naturally
occurring amino acids also illustratively include 4-hydroxyproline,
gamma.-carboxyglutamic acid, selenocysteine, desmosine,
6-N-methyllysine, epsilon.-N,N,N-trimethyllysine,
3-methylhistidine, O-phosphoserine, 5-hydroxylysine,
epsilon.-N-acetyllysine, omega.-N-methylarginine, N-acetylserine,
gamma-aminobutyric acid, citrulline, ornithine, azaserine,
homocysteine, beta.-cyanoalanine and S-adenosylmethionine.
Non-naturally occurring amino acids include phenyl glycine,
meta-tyrosine, para-amino phenylalanine, 3-(3-pyridyl)-L-alanine-,
4-(trifluoromethyl)-D-phenylalanine, and the like.
[0208] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (1), (2)
and (3) as defined above, except that R.sup.20 of Z is described in
U.S. Pat. No. 4,443,435 (incorporated by reference in its entirety)
as comprising --CH(R.sup.130)--X--C(O)--R.sup.131 wherein
[0209] X is O, S, or NR.sup.132;
[0210] R.sup.131 is [0211] (a) straight or branched chain alkyl
having from 1 to 20 carbon atoms especially methyl, ethyl,
isopropyl, t-butyl, pentyl or hexyl; [0212] (b) aryl having from 6
to 10 carbon atoms especially phenyl, substituted penyl or
naphthalene; [0213] (c) cycloalkyl having from 3 to 8 carbon atoms
especially cyclopentyl, or cyclohexyl; [0214] (d) alkenyl having
from 2-20 carbon atoms especially C.sub.2-6 alkenyl such as vinyl,
allyl, or butenyl; [0215] (e) cycloalkenyl having from 5 to 8
carbon atoms especially cyclopentenyl or cyclohexenyl; [0216] (f)
alkynyl having from 2 to 20 carbon atoms especially C.sub.2-6
alkynyl for example, ethynyl, propynyl or hexynyl; [0217] (g)
aralkyl, alkaryl, aralkenyl, aralkynyl, alkenylaryl or alkynylaryl
wherein alkyl, aryl, alkenyl and alkynyl are as previously defined;
[0218] (h) loweralkoxycarbonyl especially C.sub.1-6 alkoxycarbonyl
such as methoxycarbonyl, ethoxycarbonyl, t-butoxycarbonyl and
cyclopentoxycarbonyl; [0219] (i) carboxyalkyl or alkanoyloxyalkyl
especially carboxy-C.sub.1-6 alkyl such as formyloxymethyl and
formyloxypropyl; or C.sub.1-6 (alkylcarboxyalkyl) such as
acetoxymethyl, n-propanoyloxyethyl and pentanoyloxybutyl; [0220]
(j) saturated or unsaturated monoheterocyclic or polyheterocyclic,
or fused heterocyclic, either directly bonded to the carbonyl
function or linked thereto via an alkylene bridge, containing from
1 to 3 of any one or more of the heteroatoms N, S or O in each
heterocyclic ring thereof and each such ring being from 3- to
8-membered; and [0221] (k) mono- or polysubstituted derivatives of
the above, each of said substituents being selected from the group
consisting of lower alkyl; lower alkoxy; lower alkanoyl; lower
alkanoyloxy; halo especially bromo, chloro, or fluoro;
haloloweralkyl especially fluoro, chloro or bromoloweralkyl such as
trifluoromethyl and 1-chloropropyl; cyano; carbethoxy;
loweralkylthio, especially C.sub.1-6 loweralkylthio such as
methylthio, ethylthio and n-propylthio; nitro; carboxyl; amino;
loweralkylamino especially C.sub.1-6 alkylamino, for example,
methylamino, ethylamino and n-butylamino; diloweralkylamino
especially di(C.sub.1-6 loweralkyl)amino such as N,N-dimethylamino,
N,N-diethylamino and N,N-dihexylamino; carbamyl; loweralkylcarbamyl
especially C.sub.1-6 alkylcarbamyl such as methylcarbamyl and ethyl
carbamoyl; and R.sup.133--X--C(O)-phenyl-, wherein R.sup.133 is
hydrogen or alkyl having from 1 to 10 carbons;
[0222] R.sup.130 is hydrogen, (b) R.sup.131, lower alkanoyl, cyano,
haloloweralkyl, carbamyl, loweralkylcarbamyl, or
diloweralkylcarbamyl, --CH.sub.2ONO.sub.2, or
--CH.sub.2OCOR.sup.131;
[0223] R.sup.132 is hydrogen or lower alkyl; and further wherein
R.sup.131 and R.sup.130 may be taken together to form a ring
cyclizing moiety selected from the group consisting of:
##STR00009##
[0224] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (1), (2)
and (3) as defined above, except that R.sup.20 of Z is described in
U.S. Pat. No. 6,407,235 (incorporated by reference in its entirety)
as comprising:
[0225] a) --C(O)(CH.sub.2).sub.mC(O)OR.sup.40, wherein m is 1, 2, 3
or 4, [0226] b)
##STR00010##
[0226] wherein R.sup.41 is --N(R.sup.42)(R.sup.43) and R.sup.42 and
R.sup.43 are hydrogen or lower alkyl, or is a five or six member
heterocyclyl or heteroaryl optionally substituted by lower alkyl,
or
[0227] c)
--C(O)(CH.sub.2)NHC(O)(CH.sub.2)N(R.sup.42)(R.sup.43).
[0228] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (1), (2)
and (3) as defined above, except that R.sup.20 of Z is described in
U.S. Pat. No. 6,545,131 (incorporated by reference in its entirety)
as comprising:
CO--(CH.dbd.CH).sub.n1--(CH.sub.2).sub.n2--Ar--NH.sub.2,
--CO--(CH.sub.2).sub.n2--(CH.dbd.CH).sub.n1--Ar--NH.sub.2,
CO--(CH.sub.2).sub.n2--(CH.dbd.CH).sub.n1--CO--NH--Ar--NH.sub.2 and
CO--(CH.dbd.CH).sub.n1--(CH.sub.2).sub.n2--CO--NH--Ar--NH.sub.2 and
substituted variations thereof, where n1 and n2 are from 0 to 5, Ar
is a substituted or unsubstituted aryl group. In some preferred
embodiments, Z is CO--(CH.sub.2).sub.n3--NH.sub.2, where n3 is from
0 to 15, preferably 3-15, and also preferably 6-12. Particularly
preferred substituent groups within this class are 6-aminohexanoyl,
7-aminoheptanoyl, 8-aminooctanoyl, 9-aminononanoyl,
10-aminodecanoyl, 11-aminoundecanoyl, and 12-aminododecanoyl. These
substituents are generally synthesized from the corresponding amino
acids, 6-aminohexanoic acid, and so forth. The amino acids are
N-terminal protected by standard methods, for example Boc
protection. Dicyclohexylcarbodiimide (DCCI)-promoted coupling of
the N-terminal protected substituent to thapsigargin, followed by
standard deprotection reactions produces primary amine-containing
thapsigargin analogs.
[0229] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (1), (2)
and (3) as defined above, except that R.sup.20 of Z is described in
U.S. Pat. No. 7,115,573 (incorporated by reference in its entirety)
as comprising: [0230] (1) an oligopeptide of the formula
(AA).sub.n-AA.sup.3-AA.sup.2-AA.sup.1, wherein: each AA
independently represents an amino acid, n is 0 or 1, and when n is
1, then (AA).sub.n is AA.sup.4 which represents any amino acid,
AA.sup.3 represents isoleucine, AA.sup.2 represents any amino acid,
and AA.sup.1 represents any amino acid, [0231] (2) a stabilizing
group, and [0232] (3) optionally, a linker group not cleavable by a
trouase, such as TOP (described in greater detail below)
[0233] wherein the oligopeptide is directly linked to the
stabilizing group at a first attachment site of the oligopeptide
and the oligopeptide is directly linked to the therapeutic agent or
indirectly linked through the linker group to the therapeutic agent
at a second attachment site of the oligopeptide,
[0234] wherein the stabilizing group hinders cleavage of the
compound by enzymes present in whole blood, and
[0235] wherein the compound is cleavable by an enzyme associated
with the target cell, the enzyme associated with the target cell
being other than TOP (Thimet oligopeptidase). The compound
preferably includes an oligopeptide that is resistant to cleavage
by a trouase, particularly TOP, i.e., resistant to cleavage under
physiological conditions. The optionally present linker group that
is not cleavable by a trouase is not cleavable under physiological
conditions.
[0236] The typical orientation of these portions of the prodrug is
as follows: (stabilizing group)-(oligopeptide)-(optional linker
group)-(therapeutic agent).
[0237] Direct linkage of two portions of the prodrug means a
covalent bond exists between the two portions. The stabilizing
group and the oligopeptide are therefore directly linked via a
covalent chemical bond at the first attachment site of the
oligopeptide, typically the N-terminus of the oligopeptide. When
the oligopeptide and the therapeutic agent are directly linked then
they are covalently bound to one another at the second attachment
site of the oligopeptide. The second attachment site of the
oligopeptide is typically the C-terminus of the oligopeptide, but
may be elsewhere on the oligopeptide.
[0238] Indirect linkage of two portions of the prodrug means each
of the two portions is covalently bound to a linker group. In an
alternative embodiment, the prodrug has indirect linkage of the
oligopeptide to the therapeutic agent. Thus, typically, the
oligopeptide is covalently bound to the linker group which, in
turn, is covalently bound to the therapeutic agent.
[0239] In an alternative embodiment, the orientation of the prodrug
may be reversed so that a stabilizing group is attached to the
oligopeptide at the C-terminus and the therapeutic agent is
directly or indirectly linked to the N-terminus of the
oligopeptide. Thus, in an alternative embodiment, the first
attachment site of the oligopeptide may be the C-terminus of the
oligopeptide and the second attachment site by the oligopeptide may
be the N-terminus of the oligopeptide. The linker group may
optimally be present between the therapeutic agent and the
oligopeptide. The alternative embodiment of the prodrug of the
invention functions in the same manner as does the primary
embodiment.
[0240] The stabilizing group typically protects the prodrug from
cleavage by proteinases and peptidases present in blood, blood
serum, and normal tissue. Particularly, since the stabilizing group
caps the N-terminus of the oligopeptide, and is therefore sometimes
referred to as an N-cap or N-block, it serves to ward against
peptidases to which the prodrug may otherwise be susceptible. A
stabilizing group that hinders cleavage of the oligopeptide by
enzymes present in whole blood is chosen from the following: [0241]
(1) other than an amino acid, or [0242] (2) an amino acid that is
either (i) a non-genetically-encoded amino acid or (ii) aspartic
acid or glutamic acid attached to the N-terminus of the
oligopeptide at the .beta.-carboxyl group of aspartic acid or the
.gamma.-carboxyl group of glutamic acid.
[0243] For example, dicarboxylic (or a higher order carboxylic)
acid or a pharmaceutically acceptable salt thereof may be used as a
stabilizing group. Since chemical radicals having more than two
carboxylic acids are also acceptable as part of the prodrug, the
end group having dicarboxylic (or higher order carboxylic) acids is
an exemplary N-cap. The N-cap may thus be a monoamide derivative of
a chemical radical containing two or more carboxylic acids where
the amide is attached onto the amino terminus of the peptide and
the remaining carboxylic acids are free and uncoupled. For this
purpose, the N-cap is preferably succinic acid, adipic acid,
glutaric acid, or phthalic acid, with succinic acid and adipic acid
being most preferred. Other examples of useful N-caps in the
prodrug compound of the invention include diglycolic acid, fumaric
acid, naphthalene dicarboxylic acid, pyroglutamic acid, acetic
acid, 1- or 2-, naphthylcarboxylic acid, 1,8-naphthyl dicarboxylic
acid, aconitic acid, carboxycinnamic acid, triazole dicarboxylic
acid, gluconic acid, 4-carboxyphenyl boronic acid, a
(PEG).sub.n-analog such as polyethylene glycolic acid, butane
disulfonic acid, maleic acid, nipecotic acid, and isonipecotic
acid.
[0244] Further, a non-genetically encoded amino acid such as one of
the following may also be used as the stabilizing group:
.beta.-Alanine, Thiazolidine-4-carboxylic acid, 2-Thienylalanine,
2-Naphthylalanine, D-Alanine, D-Leucine, D-Methionine,
D-Phenylalanine, 3-Amino-3-phenylpropionic acid,
.gamma.-Aminobutyric acid, 3-amino-4,4-diphenylbutyric acid,
Tetrahydroisoquinoline-3-carboxylic acid, 4-Aminomethylbenzoic
acid, and Aminoisobutyric acid.
[0245] A linker group between the oligopeptide and the therapeutic
agent may be advantageous for reasons such as the following: 1. As
a spacer for steric considerations in order to facilitate enzymatic
release of the AA.sup.1 amino acid or other enzymatic activation
steps. 2. To provide an appropriate attachment chemistry between
the therapeutic agent and the oligopeptide. 3. To improve the
synthetic process of making the prodrug conjugate (e.g., by
pre-derivitizing the therapeutic agent or oligopeptide with the
linker group before conjugation to enhance yield or specificity.)
4. To improve physical properties of the prodrug. 5. To provide an
additional mechanism for intracellular release of the drug.
[0246] Linker structures are dictated by the required
functionality. Examples of potential linker chemistries are
hydrazide, ester, ether, and sulfhydryl Amino caproic acid is an
example of a bifunctional linker group. When amino caproic acid is
used as part of the linker group, it is not counted as an amino
acid in the numbering scheme of the oligopeptide.
[0247] The oligopeptide moiety is linked at a first attachment site
of the oligopeptide to a stabilizing group that hinders cleavage of
the oligopeptide by enzymes present in whole blood, and directly or
indirectly linked to a therapeutic agent at a second attachment
site of the oligopeptide. The linkage of the oligopeptide to the
therapeutic agent and the stabilizing group may be performed in any
order or concurrently. The resulting conjugate is tested for
cleavability by TOP. Test compounds resistant to cleavage by TOP
are selected. The resulting conjugate may also be tested for
stability in whole blood. Test compounds stable in whole blood are
selected.
[0248] The combination of oligopeptide, stabilizing group, and
optional linker of U.S. Pat. No. 7,115,573 is further described in
US 2002-0142955, also incorporated herein by reference.
[0249] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (1), (2)
and (3) as defined above, except that R.sup.20 of Z is described in
US 2004-0019017 A1 (incorporated by reference in its entirety and
which describes caspase inhibitor prodrugs), as comprising:
##STR00011##
[0250] wherein R.sup.51 is a saturated or unsaturated,
straight-chain or branched, substituted or unsubstituted alkyl of 2
to 30, preferably 2 to 24, carbon atoms;
[0251] R.sup.52 is H or a phospholipid head group, preferably
choline;
[0252] X is a direct covalent bond or a group C(O)LR.sup.53 wherein
L is a saturated or unsaturated, straight-chain or branched,
substituted or unsubstituted alkyl having from 2 to 15 carbon
atoms, which optionally includes cyclic elements, and is optionally
interrupted by one or more atoms selected from the group consisting
of oxygen, sulfur and N(R.sup.54); R.sup.53 is selected from the
group consisting of O, S and N(R.sup.54), wherein R.sup.54 is H or
a saturated or unsaturated alkyl having 1 to 6 carbon atoms.
[0253] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (1), (2)
and (3) as defined above, except that R.sup.20 of Z is the Y moiety
described in U.S. Pat. No. 7,115,573 (incorporated by reference in
its entirety).
[0254] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (1), (2)
and (3) as defined above, except that R.sup.20 of Z is described in
US 2006-0166903 A1 (incorporated by reference in its entirety, as
comprising-X-L-O--P(O)(O.sup.-)--O--CH.sub.2--CH.sub.2--N(CH.sub.3).sub.3-
.sup.+, wherein X and L are as described in US 2006-0166903A1.
[0255] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (1), (2)
and (3) as defined above, except Z is one of the cleavable prodrug
moieties described in U.S. Pat. No. 6,855,702, US 2005-0137141, and
US 2006-0135594, all hereby incorporated by reference in their
entirety.
[0256] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (1), (2)
and (3) as defined above, wherein
[0257] Cy is optionally substituted aryl, preferably optionally
substituted phenyl;
[0258] Ar is optionally substituted aryl, preferably optionally
substituted phenyl;
[0259] R.sup.x is H or OH; and
[0260] Z is --O--R.sup.20 or R.sup.21.
[0261] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (1), (2)
and (3) as defined above, wherein
[0262] Cy is optionally substituted aryl, preferably optionally
substituted phenyl;
[0263] Ar is optionally substituted aryl, preferably optionally
substituted phenyl;
[0264] Rx is H or OH; and
[0265] Z is --O--R.sup.20 or R.sup.21, wherein
[0266] R.sup.20 is
--C(O)--C[(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13) or
--C(O)--R.sup.10.
[0267] In a preferred embodiment of the present invention, the
prodrugs of inhibitors of histone deacetylase comprise those of
formula (2).
[0268] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (2) as
defined above, wherein
[0269] Cy is optionally substituted aryl, preferably optionally
substituted phenyl;
[0270] Ar is optionally substituted aryl, preferably optionally
substituted phenyl;
[0271] Rx is H or OH; and
[0272] Z is --O--R.sup.20 or R.sup.21.
[0273] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (2) as
defined above, wherein
[0274] Cy is optionally substituted aryl, preferably optionally
substituted phenyl;
[0275] Ar is optionally substituted aryl, preferably optionally
substituted phenyl;
[0276] Rx is H or OH; and
[0277] Z is --O--R.sup.20 or R.sup.21, wherein
[0278] R.sup.20 is
--C(O)--C[(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13) or
--C(O)--R.sup.10.
[0279] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (2) as
defined above, wherein [0280] Cy is optionally substituted aryl,
preferably optionally substituted phenyl, wherein the substituents
are preferably selected from the group consisting of --CF.sub.3,
halo, heterocyclyl and fused heterocyclyl; [0281] L.sup.2 is
saturated C.sub.3alkyl or C.sub.4alkyl, preferably unsubstituted;
[0282] Ar is optionally substituted aryl, preferably optionally
substituted phenyl; [0283] Y.sup.2 is C.sub.1alkyl or C.sub.2alkyl,
preferably C.sub.1alkyl, optionally substituted; [0284] Rx is H or
OH, preferably H; [0285] Z is --O--R.sup.20 or R.sup.21; [0286]
R.sup.20 is --C(O)--C[(R.sup.10)(R.sup.10')].sub.1-4--NH(R) or
--C(O)--R.sup.10; [0287] each R.sup.10 is independently selected
from the group consisting of H, optionally substituted alkyl,
optionally substituted -alkylphenyl, optionally substituted
-alkylheteroaryl and optionally substituted heteroaryl; [0288] each
R.sup.10' is independently H or alkyl; or [0289] R.sup.10 and
R.sup.10' together with the atom to which they are attached form a
C.sub.3 or C.sub.4spirocycloalkyl, preferably a
C.sub.3spirocycloalkyl; [0290] R.sup.13 is selected from the group
consisting of H,
--C(O)--CH[N(R.sup.10)(R.sup.10')]-C.sub.1-C.sub.6alkyl-N(R.sup.10)(R.sup-
.10'), --C(O)-heteroaryl, --C(O)-aryl,
--C(O)--CH[N(R.sup.10)(R.sup.10')]-C.sub.1-C.sub.6alkyl,
--C(O)--CH[N(R.sup.10)(R.sup.10')]-C.sub.1-C.sub.6alkyl-aryl and
--C(O)--CH[N(R.sup.10)(R.sup.10')]-C.sub.1-C.sub.6alkyl-heteroaryl;
and [0291] R.sup.21 is amino acid-R.sup.13 (preferably the amino
acid is lysine or arginine).
[0292] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (2) as
defined above, wherein [0293] Cy is optionally substituted aryl,
preferably optionally substituted phenyl, wherein the substituents
are preferably selected from the group consisting of --CF.sub.3,
halo, heterocyclyl and fused heterocyclyl; [0294] L.sup.2 is
saturated C.sub.3alkyl or C.sub.4alkyl, preferably unsubstituted;
[0295] Ar is optionally substituted aryl, preferably optionally
substituted phenyl; [0296] Y.sup.2 is C.sub.1alkyl or C.sub.2alkyl,
preferably C.sub.1alkyl, optionally substituted; [0297] Rx is H or
OH, preferably H; [0298] Z is --O--R.sup.20; [0299] R.sup.20 is
--C(O)--C[(R.sup.10)(R.sup.10')].sub.1-4--NH(R.sup.13), preferably
--C(O)--C[(R.sup.10)(R.sup.10')].sub.1-2--NH(R.sup.13), more
preferably --C(O)--C[(R.sup.10)(R.sup.10')]--NH(R.sup.13); [0300]
each R.sup.10 is independently selected from the group consisting
of H, optionally substituted alkyl and optionally substituted
-alkylphenyl; [0301] each R.sup.10' is independently H or alkyl; or
[0302] R.sup.10 and R.sup.10' together with the atom to which they
are attached form a C.sub.3 or C.sub.4spirocycloalkyl, preferably a
C.sub.3spirocycloalkyl; and [0303] R.sup.13 is H.
[0304] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (2) as
defined above, wherein [0305] Cy is optionally substituted aryl,
preferably optionally substituted phenyl, wherein the substituents
are preferably selected from the group consisting of --CF.sub.3,
halo, heterocyclyl and fused heterocyclyl; [0306] L.sup.2 is
saturated C.sub.3alkyl or C.sub.4alkyl, preferably unsubstituted;
[0307] Ar is optionally substituted aryl, preferably optionally
substituted phenyl; [0308] Y.sup.2 is C.sub.1alkyl or C.sub.2alkyl,
preferably C.sub.1alkyl, optionally substituted; [0309] Rx is H or
OH, preferably H; [0310] Z is R.sup.21; [0311] R.sup.21 is amino
acid-R.sup.13 (preferably the amino acid is lysine or arginine);
and [0312] R.sup.13 is H.
[0313] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (2) as
defined above, wherein [0314] Cy is optionally substituted aryl,
preferably optionally substituted phenyl, wherein the substituents
are preferably selected from the group consisting of --CF.sub.3,
halo, heterocyclyl and fused heterocyclyl; [0315] L.sup.2 is
saturated C.sub.3alkyl or C.sub.4alkyl, preferably unsubstituted;
[0316] Ar is optionally substituted aryl, preferably optionally
substituted phenyl; [0317] Y.sup.2 is C.sub.1alkyl or C.sub.2alkyl,
preferably C.sub.1alkyl, optionally substituted; [0318] Rx is H or
OH, preferably H; [0319] Z is --O--R.sup.20; [0320] R.sup.20 is
--C(O)--R.sup.10; and [0321] R.sup.10 is selected from the group
consisting of optionally substituted alkyl, optionally substituted
-alkylphenyl, optionally substituted -alkylheteroaryl and
optionally substituted heteroaryl.
[0322] In other embodiments, the prodrugs of inhibitors of histone
deacetylase of the invention comprise those of formulae (2) as
defined above, wherein [0323] Cy is optionally substituted aryl,
preferably optionally substituted phenyl, wherein the substituents
are preferably selected from the group consisting of --CF.sub.3,
halo, heterocyclyl and fused heterocyclyl; [0324] L.sup.2 is
saturated C.sub.3alkyl or C.sub.4alkyl, preferably unsubstituted;
[0325] Ar is optionally substituted aryl, preferably optionally
substituted phenyl; [0326] Y.sup.2 is C.sub.1alkyl or C.sub.2alkyl,
preferably C.sub.1alkyl, optionally substituted; [0327] Rx is H or
OH, preferably H; [0328] Z is --O--R.sup.20; [0329] R.sup.20 is
--C(O)--C[(R.sup.10)(R.sup.10')]-NH(R.sup.13), wherein R.sup.10 and
R.sup.10' together with the atom to which they are attached form a
C.sub.3 or C.sub.4spirocycloalkyl, preferably a
C.sub.3spirocycloalkyl; and [0330] R.sup.13 is selected from the
group consisting of H,
--C(O)--CH[N(R.sup.10)(R.sup.10')]-C.sub.1-C.sub.6alkyl-N(R.sup.10)(R.sup-
.10'), --C(O)-heteroaryl, --C(O)-aryl,
--C(O)--CH[N(R.sup.10)(R.sup.10')]-C.sub.1-C.sub.6alkyl,
--C(O)--CH[N(R.sup.10)(R.sup.10')]-C.sub.1-C.sub.6alkyl-aryl and
--C(O)--CH[N(R.sup.10)(R.sup.10')]-C.sub.1-C.sub.6alkyl-heteroaryl,
wherein R.sup.10 and R.sup.10' are each independently selected from
H and C.sub.1-C.sub.6alkyl, preferably H.
[0331] Preferred prodrugs of the invention include those in Table
A:
TABLE-US-00001 TABLE A ##STR00012## ##STR00013## ##STR00014##
##STR00015## ##STR00016## ##STR00017## ##STR00018## ##STR00019##
##STR00020## ##STR00021## ##STR00022## ##STR00023## ##STR00024##
##STR00025## ##STR00026## ##STR00027## ##STR00028## ##STR00029##
##STR00030## ##STR00031## ##STR00032## ##STR00033## ##STR00034##
##STR00035## ##STR00036## ##STR00037##
[0332] Preferred prodrug compounds of the invention are cleavable
(e.g., hydrolysable) in mammalian and/or fungal pathogen cells into
compounds (cleavage products) in which Z in formulae (1), (2), and
(3) is --OH. Such cleavage products are active histone deacetylase
inhibitors. Thus, according to another aspect, the invention
provides compounds of formulae (1), (2), and (3) as defined above
(and pharmaceutically acceptable salts thereof) with the exception
that Z is --OH. Among the preferred cleavage compounds are those
with structure:
##STR00038##
[0333] Preferred cleavage products of the prodrug compounds of the
invention include those in Table A in which Z is --OH.
[0334] All compounds of the invention, whether prodrug or
corresponding cleavage product, can be racemic or
diastereomerically or enantiomerically enriched. In addition,
compounds of the invention, whether prodrug or corresponding
cleavage product, can be in the form of a hydrate, solvate,
pharmaceutically acceptable salt, and/or complex.
Synthesis
[0335] Compounds of formula Cy-L.sup.1-Ar-Y.sup.1--C(O)--NH--O--H,
wherein L.sup.1 is --S(O).sub.2NH--, preferably may be prepared
according to the synthetic routes depicted in Schemes 1-5.
Accordingly, in certain preferred embodiments, compounds I are
preferably prepared according to the general synthetic route
depicted in Scheme 1. Thus, a sulfonyl chloride (II) is treated
with an amine (III) in a solvent such as methylene chloride in the
presence of an organic base such as triethylamine Treatment of the
crude product with a base such as sodium methoxide in an alcoholic
solvent such as methanol effects cleavage of any dialkylated
material and affords the sulfonamide (IV). Hydrolysis of the ester
function in IV can be effected by treatment with a hydroxide base,
such as lithium hydroxide, in a solvent mixture such as
tetrahydrofuran and methanol to afford the corresponding acid
(V).
##STR00039##
[0336] In some embodiments, conversion of the acid V to the
hydroxamic acid I may be accomplished by coupling V with a
protected hydroxylamine, such as tetrahydropyranylhydroxylamine
(NH.sub.2OTHP), to afford the protected hydroxamate VI, followed by
acidic hydrolysis of VI to provide the hydroxamic acid I. The
coupling reaction is preferably accomplished with the coupling
reagent dicyclohexylcarbodiimide (DCC) in a solvent such as
methylene chloride (Method A) or with the coupling reagent
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide in presence of
N-hydroxy benzotriazole in an aprotic solvent such as
dimethylformamide (Method D). Other coupling reagents are known in
the art and may also be used for this reaction. Hydrolysis of VI is
preferably effected by treatment with an organic acid such as
camphorsulfonic acid in a protic solvent such as methanol.
[0337] Alternatively, in some other embodiments, acid V is
converted to the corresponding acid chloride, preferably by
treatment with oxalic chloride, followed by the addition of a
protected hydroxylamine such as O-trimethylsilylhydroxylamine in a
solvent such as methylene chloride, which then provides the
hydroxylamine I upon workup (Method C).
[0338] In still other embodiments, the ester IV is preferably
treated with hydroxylamine in a solvent such as methanol in the
presence of a base such as sodium methoxide to furnish the
hydroxylamine I directly (Method B).
##STR00040##
[0339] Compounds of formula X and XIV preferably are prepared
according to the general procedure outlined in Scheme 2. Thus, an
aminoaryl halide (VII) is treated with a sulfonyl chloride in
presence of a base such as triethylamine, followed by treatment
with an alkoxide base, to furnish the sulfonamide VIII. One of
skill in the art will recognize that reverse sulfonamide analogs
can be readily prepared by an analogous procedure, treating a
haloarenesulfonyl halide with an arylamine
[0340] Compound VIII is coupled with a terminal acetylene or
olefinic compound in the presence of a palladium catalyst such as
tetrakis(triphenylphosphine)palladium(0) in a solvent such as
pyrrolidine to afford IX.
[0341] Oxidation of the compound of formula IX (X=CH.sub.2OH),
followed by homologation of the resulting aldehyde using a Wittig
type reagent such as carbethoxymethylenetriphenylphosphorane in a
solvent such as acetonitrile, gives the compound of formula XI.
Basic hydrolysis of XI, such as by treatment with lithium hydroxide
in a mixture of THF and water, provides the acid XII. Hydrogenation
of XII may preferably be performed over a palladium catalyst such
as Pd/C in a protic solvent such as methanol to afford the
saturated acid XIII Coupling of the acid XIII with an O-protected
hydroxylamine such as O-tetrahydropyranylhydroxylamine is effected
by treatment with a coupling reagent such as
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide in the presence of
N-hydroxybenzotriazole (HOBT), or N,N-dicyclohexylcarbodiimide
(DCC), in a solvent such as DMF, followed by deprotection to
furnish the compound of general formula XIV.
[0342] The acid IX, wherein X=COOH, may be coupled directly with an
O-protected hydroxylamine such as O-tetrahydropyranylhydroxylamine,
followed by deprotection of the hydroxy protecting group to furnish
the hydroxamic acid X.
[0343] Compounds of formula Cy-L.sup.1-Ar--Y.sup.1--C(O)--NH--O--H,
wherein L.sup.1 is --C(O)NH--, preferably may be prepared according
to the synthetic routes analogous to those depicted in Schemes 1-2,
substituting acid chloride starting materials for the sulfonyl
chloride starting materials in those Schemes.
##STR00041##
[0344] Compounds of the formula
Cy-L.sup.2-Ar-Y.sup.2--C(O)--NH--O--H are preferably prepared
according to the synthetic routes outlined in Schemes 3-5.
Accordingly, in certain preferred embodiments, compounds of
formulae XIX and XXI (L.sup.2=--C(O)--CH.dbd.CH-- or
--C(O)--CH.sub.2CH.sub.2--) preferably are prepared according to
the route described in Scheme 3. Thus, a substituted aryl
acetophenone (XV) is treated with an aryl aldehyde (XVI) in a
protic solvent such as methanol in the presence of a base such as
sodium methoxide to afford the enone XVII.
[0345] The acid substituent of XVII (R=H) is coupled with an
O-protected hydroxylamine such as O-tetrahydropyranylhydroxylamine
(R.sub.1=tetrahydropyranyl) to afford the
O-protected-N-hydroxybenzamide XVIII. The coupling reaction is
preferably performed by treating the acid and hydroxylamine with
dicyclohexylcarbodiimide in a solvent such as methylene chloride or
with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide in the presence
of N-hydroxybenzotriazole in a solvent such as dimethylformamide.
Other coupling reagents are known in the art and may also be used
in this reaction. O-Deprotection is accomplished by treatment of
XVIII with an acid such as camphorsulfonic acid in a solvent such
as methanol to afford the hydroxamic acid XIX
(L.sup.2=--C(O)--CH.dbd.CH--).
[0346] Saturated compounds of formula XXI
(L.sup.2=--C(O)--CH.sub.2CH.sub.2--) are preferably prepared by
hydrogenation of XVII (R=Me) over a palladium catalyst, such as 10%
Pd/C, in a solvent such as methanol-tetrahydrofuran. Basic
hydrolysis of the resultant product XIX with lithium hydroxide,
followed by N-hydroxy amide formation and acid hydrolysis as
described above, then affords the hydroxamic acid XXI.
##STR00042##
[0347] Compounds of formula XXVI (L.sup.2=--(CH.sub.2).sub.o+2--)
are preferably prepared by the general procedures described in
Schemes 4 and 5. Thus, in some embodiments, a terminal olefin
(XXII) is coupled with an aryl halide (XXIII) in the presence of a
catalytic amount of a palladium source, such as palladium acetate
or tris(dibenzylideneacetone)dipalladium(0), a phosphine, such as
triphenylphosphine, and a base, such as triethylamine, in a solvent
such as acetonitrile to afford the coupled product XXIV.
Hydrogenation, followed by N-hydroxyamide formation and acid
hydrolysis, as described above, affords the hydroxamic acid
XXVI.
##STR00043##
[0348] Alternatively, in some other embodiments, a phosphonium salt
of formula XXVII is treated with an aryl aldehyde of formula XXVIII
in the presence of base, such as lithium hexamethyldisilazide, in a
solvent, such as tetrahydrofuran, to produce the compound XXIV.
Hydrogenation, followed by N-hydroxyamide formation and acidic
hydrolysis, then affords the compounds XXVI.
##STR00044##
[0349] Compounds of formula Cy-L-Ar-Y--C(O)--NH--Z, wherein L is
L.sup.1 or L.sup.2, Y is Y.sup.1 or Y.sup.2, and Z is anilinyl or
pyridyl, are preferably prepared according to synthetic routes
outlined in Scheme 6. An acid of formula Cy-L-Ar-Y--C(O)--OH
(XXIX), prepared by one of the methods shown in Schemes 1-5, is
converted to the corresponding acid chloride XXX according to
standard methods, e.g., by treatment with sodium hydride and oxalyl
chloride. Treatment of XXX with 2-aminopyridine and a tertiary base
such as N-methylmorpholine, preferably in dichloromethane at
reduced temperature, then affords the pyridyl amide XXXI. In a
similar fashion, the acid chloride XXX may be treated with
1,2-phenylenediamine to afford the anilinyl amide XXXII.
Alternatively, the acid chloride XXX may be treated with a
mono-protected 1,2-phenylenediamine, such as
2-(t-BOC-amino)aniline, followed by deprotection, to afford
XXXII.
[0350] In another alternative procedure, the acid XXIX may be
activated by treatment with carbonyldiimidazole (CDI), followed by
treatment with 1,2-phenylenediamine and trifluoroacetic acid to
afford the anilinyl amide XXXII.
##STR00045##
[0351] Compounds of formula XXXVIII (L.sup.2=--C(O)-alkylene-)
preferably are prepared according to the general procedure depicted
in Scheme 7. Thus, Aldol condensation of ketone XXXIII (R.sub.1=H
or alkyl) with aldehyde XXXIV affords the adduct XXXV. The adduct
XXXV may be directly converted to the corresponding hydroxamic acid
XXXVI, or may first undergo hydrogenation to afford the saturated
compound XXVII and then be converted to the hydroxamic acid
XXXVIII.
##STR00046##
[0352] Compounds of formula Cy-L.sup.2-Ar-Y.sup.2--C(O)--NH--O--H,
wherein one of the carbon atoms in L.sup.2 is replaced with S,
S(O), or S(O).sub.2 preferably are prepared according to the
general procedure outlined in Scheme 8. Thus, thiol XXXIX is added
to olefin XL to produce XLI. The reaction is preferably conducted
in the presence of a radical initiator such as
2,2'-azobisisobutyronitrile (AIBN) or
1,1'-azobis(cyclohexanecarbonitrile) (VAZO.TM.). Sulfide oxidation,
preferably by treatment with m-chloroperbenzoic acid (mCPBA),
affords the corresponding sulfone, which is conveniently isolated
after conversion to the methyl ester by treatment with
diazomethane. Ester hydrolysis then affords the acid XLII, which is
converted to the hydroxamic acid XLIII according to any of the
procedures described above. The sulfide XLI also may be converted
directly to the corresponding hydroxamic acid XLIV, which then may
be selectively oxidized to the sulfoxide XLV, for example, by
treatment with hydrogen peroxide and tellurium dioxide.
[0353] Alternatively, compounds of
Cy-L.sup.2-Ar-Y.sup.2--C(O)--NH--O--H can be prepared according to
Scheme 9. In Scheme 9, haloaryl acetic acid XLVI is esterified, by,
for example, treatment with HCl in dioxane in the presence of an
alcohol such as methanol, to afford acetate XLVII. Paladium
coupling of acetate XLVII with alkyne XLVIII with, for example
(Ph.sub.3P).sub.4Pd in DME and diethylamine in the presence of Cut,
produces XLVIX, which is subsequently reduced under H.sub.2 and,
for example, Pd/C in methanol, to afford XLVX. N-hydroxyamide
formation and acid hydrolysis, as described above, then leads to
XLVXI.
##STR00047##
[0354] Compounds of formulas (1)-(3) can be prepared as depicted in
Scheme 10. XLVXI is treated with an amino acid XLVXII under
standard peptide coupling conditions, such as, for example, EDC and
HOBt in DMF, to afford the protected acetamide XLVXIII, which is
subsequently deprotected to yield the prodrug XLVXIV.
##STR00048##
[0355] Other compounds of formula (1)-(3) can be prepared by
methods known by those skilled in the art. Examples of such methods
can be found in U.S. Pat. Nos. 4,443,435; 6,407,235; 6,545,131;
6,855,702; 7,115,573; United States Patent Application Nos. US
2002-0142955, US 2004-0019017, US 2005-0137141, US 2006-0135594, US
2006-0166903 and international publication WO 2005/097747, all of
which are incorporated herein by reference.
Pharmaceutical Compositions
[0356] In a second aspect, the invention provides pharmaceutical
compositions comprising a prodrug of an inhibitor of histone
deacetylase represented by any one of formulae (1)-(3) and a
pharmaceutically acceptable carrier, excipient, or diluent.
Compounds of the invention (whether a prodrug or a hydrolzyation
product) or compositions thereof may be formulated by any method
well known in the art and may be prepared for administration by any
route, including, without limitation, parenteral, oral, sublingual,
transdermal, topical, intranasal, intratracheal, or intrarectal. In
certain preferred embodiments, compounds of the invention (whether
a prodrug or a hydrolzyation product) or compositions thereof are
administered intravenously in a hospital setting. In certain other
preferred embodiments, administration may preferably be by the oral
route.
[0357] The characteristics of the carrier will depend on the route
of administration. As used herein, the term "pharmaceutically
acceptable" means a non-toxic material that is compatible with a
biological system such as a cell, cell culture, tissue, or
organism, and that does not interfere with the effectiveness of the
biological activity of the active ingredient(s). Thus, compositions
according to the invention may contain, in addition to the
inhibitor, diluents, fillers, salts, buffers, stabilizers,
solubilizers, and other materials well known in the art. The
preparation of pharmaceutically acceptable formulations is
described in, e.g., Remington's Pharmaceutical Sciences, 18th
Edition, ed. A. Gennaro, Mack Publishing Co., Easton, Pa.,
1990.
Inhibition of Histone Deacetylase
[0358] In a third aspect, the invention provides a method of
inhibiting histone deacetylase in a cell, comprising contacting a
cell in which inhibition of histone deacetylase is desired with a
prodrug of an inhibitor of histone deacetylase according to any of
formulas (1)-(3).
[0359] Measurement of the enzymatic activity of a histone
deacetylase can be achieved using known methodologies. For example,
Yoshida et al., J. Biol. Chem., 265: 17174-17179 (1990), describes
the assessment of histone deacetylase enzymatic activity by the
detection of acetylated histones in trichostatin A treated cells.
Taunton et al., Science, 272: 408-411 (1996), similarly describes
methods to measure histone deacetylase enzymatic activity using
endogenous and recombinant HDAC-1. Both of these references are
hereby incorporated by reference in their entirety.
[0360] In some preferred embodiments, the histone deacetylase
inhibitor interacts with and reduces the activity of all histone
deacetylases in the cell. In some other preferred embodiments
according to this aspect of the invention, the histone deacetylase
inhibitor interacts with and reduces the activity of fewer than all
histone deacetylases in the cell. In certain preferred embodiments,
the inhibitor interacts with and reduces the activity of one
histone deacetylase (e.g., HDAC-1), but does not interact with or
reduce the activities of other histone deacetylases (e.g., HDAC-2,
HDAC-3, HDAC-4, HDAC-5, HDAC-6, HDAC-7, HDAC-8, HDAC-9, HDAC-10,
and HDAC-11). As discussed below, certain particularly preferred
histone deacetylase inhibitors are those that interact with and
reduce the enzymatic activity of a histone deacetylase that is
involved in tumorigenesis. Certain other preferred histone
deacetylase inhibitors interact with and reduce the enzymatic
activity of a fungal histone deacetylase.
[0361] Preferably, the method according to the third aspect of the
invention causes an inhibition of cell proliferation of the
contacted cells. The phrase "inhibiting cell proliferation" is used
to denote an ability of an inhibitor of histone deacetylase to
retard the growth of cells contacted with the inhibitor as compared
to cells not contacted. An assessment of cell proliferation can be
made by counting contacted and non-contacted cells using a Coulter
Cell Counter (Coulter, Miami, Fla.) or a hemacytometer, or other
appropriate method (which may depend on the cell type being
counted) known to those of skill in the art. Where the cells are in
a solid growth (e.g., a solid tumor or organ), such an assessment
of cell proliferation can be made by measuring the growth with
calipers and comparing the size of the growth of contacted cells
with non-contacted cells.
[0362] Preferably, growth of cells contacted with the prodrug of
the inhibitor is retarded by at least 50% as compared to growth of
non-contacted cells. More preferably, cell proliferation is
inhibited by 100% (i.e., the contacted cells do not increase in
number). Most preferably, the phrase "inhibiting cell
proliferation" includes a reduction in the number or size of
contacted cells, as compared to non-contacted cells. Thus, a
cleavage (e.g., hydrolyzation) product of a prodrug of an inhibitor
of histone deacetylase according to the invention that inhibits
cell proliferation in a contacted cell may induce the contacted
cell to undergo growth retardation, to undergo growth arrest, to
undergo programmed cell death (i.e., to apoptose), or to undergo
necrotic cell death.
[0363] The cell proliferation inhibiting ability of the histone
deacetylase inhibitors according to the invention allows the
synchronization of a population of asynchronously growing cells.
For example, the hydrolzyation products of the prodrugs of histone
deacetylase inhibitors of the invention may be used to arrest a
population of non-neoplastic cells grown in vitro in the G1 or G2
phase of the cell cycle. Such synchronization allows, for example,
the identification of gene and/or gene products expressed during
the G1 or G2 phase of the cell cycle. Such a synchronization of
cultured cells may also be useful for testing the efficacy of a new
transfection protocol, where transfection efficiency varies and is
dependent upon the particular cell cycle phase of the cell to be
transfected. Use of the prodrugs of histone deacetylase inhibitors
of the invention allows the synchronization of a population of
cells, thereby aiding detection of enhanced transfection
efficiency.
[0364] In some preferred embodiments, the contacted cell is a
neoplastic cell. The term "neoplastic cell" is used to denote a
cell that shows aberrant cell growth. Preferably, the aberrant cell
growth of a neoplastic cell is increased cell growth. A neoplastic
cell may be a hyperplastic cell, a cell that shows a lack of
contact inhibition of growth in vitro, a benign tumor cell that is
incapable of metastasis in vivo, or a cancer cell that is capable
of metastasis in vivo and that may recur after attempted removal.
The term "tumorigenesis" is used to denote the induction of cell
proliferation that leads to the development of a neoplastic growth.
In some embodiments, the cleavage product of a prodrug of a histone
deacetylase inhibitor of the invention induces cell differentiation
in the contacted cell. Thus, a neoplastic cell, when contacted with
a prodrug of an inhibitor of histone deacetylase of the invention
may be induced to differentiate, resulting in the production of a
daughter cell that is phylogenetic ally more advanced than the
contacted cell. In certain other preferred embodiments, the
contacted cell is a fungal cell.
[0365] In some preferred embodiments, the contacted cell is in an
animal Thus, the invention provides a method for treating a cell
proliferative disease or condition in an animal, or treating a
fungal infection, comprising administering to an animal in need of
such treatment a therapeutically effective amount of a prodrug of a
histone deacetylase inhibitor of the invention. Preferably, the
animal is a mammal, more preferably a domesticated mammal. Most
preferably, the animal is a human.
[0366] The term "cell proliferative disease or condition" is meant
to refer to any condition characterized by aberrant cell growth,
preferably abnormally increased cellular proliferation. Examples of
such cell proliferative diseases or conditions include, but are not
limited to, cancer, restenosis, and psoriasis. In particularly
preferred embodiments, the invention provides a method for
inhibiting neoplastic cell proliferation in an animal comprising
administering to an animal having at least one neoplastic cell
present in its body a therapeutically effective amount of a prodrug
of a histone deacetylase inhibitor of the invention.
[0367] It is contemplated that some cleavage products of the
prodrugs of the invention have inhibitory activity against a
histone deacetylase from a protozoal source. Thus, the invention
also provides a method for treating or preventing a protozoal
disease or infection, comprising administering to an animal in need
of such treatment a therapeutically effective amount of a prodrug
of a histone deacetylase inhibitor of the invention. Preferably the
animal is a mammal, more preferably a human. Preferably, the
histone deacetylase inhibitor used according to this embodiment of
the invention inhibits a protozoal histone deacetylase to a greater
extent than it inhibits mammalian histone deacetylases,
particularly human histone deacetylases.
[0368] The present invention further provides a method for treating
a fungal disease or infection comprising administering to an animal
in need of such treatment a therapeutically effective amount of a
prodrug of a histone deacetylase inhibitor of the invention.
Preferably the animal is a mammal, more preferably a human.
Preferably, the histone deacetylase inhibitor used according to
this embodiment of the invention inhibits a fungal histone
deacetylase to a greater extent than it inhibits mammalian histone
deacetylases, particularly human histone deacetylases.
[0369] The term "therapeutically effective amount" is meant to
denote a dosage sufficient to cause inhibition of histone
deacetylase activity in the cells of the subject, or a dosage
sufficient to inhibit cell proliferation or to induce cell
differentiation in the subject. Administration may be by any route,
including, without limitation, parenteral, oral, sublingual,
transdermal, topical, intranasal, intratracheal, or intrarectal. In
certain particularly preferred embodiments, prodrugs of the
invention are administered intravenously in a hospital setting. In
certain other preferred embodiments, administration may preferably
be by the oral route.
[0370] When administered systemically, the prodrug of an histone
deacetylase inhibitor is preferably administered at a sufficient
dosage to attain a blood level of the inhibitor from about 0.01
.mu.M to about 100 .mu.M, more preferably from about 0.05 .mu.M to
about 50 .mu.M, still more preferably from about 0.1 .mu.M to about
25 .mu.M, and still yet more preferably from about 0.5 .mu.M to
about 25 .mu.M. For localized administration, much lower
concentrations than this may be effective, and much higher
concentrations may be tolerated. One of skill in the art will
appreciate that the dosage of histone deacetylase inhibitor
necessary to produce a therapeutic effect may vary considerably
depending on the tissue, organ, or the particular animal or patient
to be treated.
[0371] In certain preferred embodiments of the third aspect of the
invention, the method further comprises contacting the cell with an
antisense oligonucleotide that inhibits the expression of a histone
deacetylase. The combined use of a nucleic acid level inhibitor
(i.e., antisense oligonucleotide) and a protein level inhibitor
(i.e., inhibitor of histone deacetylase enzyme activity) results in
an improved inhibitory effect, thereby reducing the amounts of the
inhibitors required to obtain a given inhibitory effect as compared
to the amounts necessary when either is used individually.
Antisense oligonucleotides according to this aspect of the
invention, when directed to mammalian HDAC, are complementary to
regions of RNA or double-stranded DNA that encode HDAC-1, HDAC-2,
HDAC-3, HDAC-4, HDAC-5, HDAC-6, HDAC7, HDAC-8, HDAC-9, HDAC-10
and/or HDAC-11.
[0372] For purposes of the invention, the term "oligonucleotide"
includes polymers of two or more deoxyribonucleosides,
ribonucleosides, or 2'-O-substituted ribonucleoside residues, or
any combination thereof. Preferably, such oligonucleotides have
from about 6 to about 100 nucleoside residues, more preferably from
about 8 to about 50 nucleoside residues, and most preferably from
about 12 to about 30 nucleoside residues. The nucleoside residues
may be coupled to each other by any of the numerous known
internucleoside linkages. Such internucleoside linkages include
without limitation phosphorothioate, phosphorodithioate,
alkylphosphonate, alkylphosphonothioate, phosphotriester,
phosphoramidate, siloxane, carbonate, carboxymethylester,
acetamidate, carbamate, thioether, bridged phosphoramidate, bridged
methylene phosphonate, bridged phosphorothioate and sulfone
internucleoside linkages. In certain preferred embodiments, these
internucleoside linkages may be phosphodiester, phosphotriester,
phosphorothioate, or phosphoramidate linkages, or combinations
thereof. The term oligonucleotide also encompasses such polymers
having chemically modified bases or sugars and/or having additional
substituents, including without limitation lipophilic groups,
intercalating agents, diamines and adamantane. For purposes of the
invention the term "2'-O-substituted" means substitution of the 2'
position of the pentose moiety with an --O-lower alkyl group
containing 1-6 saturated or unsaturated carbon atoms, or with an
--O-aryl or allyl group having 2-6 carbon atoms, wherein such
alkyl, aryl or allyl group may be unsubstituted or may be
substituted, e.g., with halo, hydroxy, trifluoromethyl, cyano,
nitro, acyl, acyloxy, alkoxy, carboxyl, carbalkoxyl, or amino
groups; or such 2' substitution may be with a hydroxy group (to
produce a ribonucleoside), an amino or a halo group, but not with a
2'-H group. The term "oligonucleotide" also encompasses linked
nucleic acid and peptide nucleic acid.
[0373] Particularly preferred antisense oligonucleotides utilized
in this aspect of the invention include chimeric oligonucleotides
and hybrid oligonucleotides.
[0374] For purposes of the invention, a "chimeric oligonucleotide"
refers to an oligonucleotide having more than one type of
internucleoside linkage. One preferred example of such a chimeric
oligonucleotide is a chimeric oligonucleotide comprising a
phosphorothioate, phosphodiester or phosphorodithioate region,
preferably comprising from about 2 to about 12 nucleotides, and an
alkylphosphonate or alkylphosphonothioate region (see e.g.,
Pederson et al. U.S. Pat. Nos. 5,635,377 and 5,366,878).
Preferably, such chimeric oligonucleotides contain at least three
consecutive internucleoside linkages selected from phosphodiester
and phosphorothioate linkages, or combinations thereof.
[0375] For purposes of the invention, a "hybrid oligonucleotide"
refers to an oligonucleotide having more than one type of
nucleoside. One preferred example of such a hybrid oligonucleotide
comprises a ribonucleotide or 2'-O-substituted ribonucleotide
region, preferably comprising from about 2 to about 12
2'-O-substituted nucleotides, and a deoxyribonucleotide region.
Preferably, such a hybrid oligonucleotide will contain at least
three consecutive deoxyribonucleosides and will also contain
ribonucleosides, 2'-O-substituted ribonucleosides, or combinations
thereof (see e.g., Metelev and Agrawal, U.S. Pat. No.
5,652,355).
[0376] The exact nucleotide sequence and chemical structure of an
antisense oligonucleotide utilized in the invention can be varied,
so long as the oligonucleotide retains its ability to inhibit
expression of the gene of interest. This is readily determined by
testing whether the particular antisense oligonucleotide is active
by quantitating the mRNA encoding a product of the gene, or in a
Western blotting analysis assay for the product of the gene, or in
an activity assay for an enzymatically active gene product, or in a
soft agar growth assay, or in a reporter gene construct assay, or
an in vivo tumor growth assay, all of which are described in detail
in this specification or in Ramchandani et al. (1997) Proc. Natl.
Acad. Sci. USA 94: 684-689.
[0377] Antisense oligonucleotides utilized in the invention may
conveniently be synthesized on a suitable solid support using well
known chemical approaches, including H-phosphonate chemistry,
phosphoramidite chemistry, or a combination of H-phosphonate
chemistry and phosphoramidite chemistry (i.e., H-phosphonate
chemistry for some cycles and phosphoramidite chemistry for other
cycles). Suitable solid supports include any of the standard solid
supports used for solid phase oligonucleotide synthesis, such as
controlled-pore glass (CPG) (see, e.g., Pon, R.T. (1993) Methods in
Molec. Biol. 20: 465-496).
[0378] Particularly, preferred oligonucleotides have nucleotide
sequences of from about 13 to about 35 nucleotides which include
the nucleotide sequences shown in Tables 1-3. Yet additional
particularly preferred oligonucleotides have nucleotide sequences
of from about 15 to about 26 nucleotides of the nucleotide
sequences shown in Tables 1-3.
TABLE-US-00002 TABLE 1 SEQ ID TARGET NO. SEQUENCE (**) 1 5'-GAG ACA
GCA GCA CCA GCG GG-3' 17-36 2 5'-ATG ACC GAG TGG GAG ACA GC-3'
21-49 3 5'-GGA TGA CCG AGT GGG AGA CA-3' 31-50 4 5'-CAG GAT GAC CGA
GTG GGA GA-3' 33-52 5 5'-TGT GTT CTC AGG ATG ACC GA-3' 41-60 6
5'-GAG TGA CAG AGA CGC TCA GG-3' 62-81 7 5'-TTC TGG CTT CTC CTC CTT
GG-3' 1504-1523 8 5'-CTT GAC CTC CTC CTT GAC CC-3' 1531-1550 9
5'-GGA AGC CAG AGC TGG AGA GG-3' 1565-1584 10 5'-GAA ACG TGA GGG
ACT CAG CA-3' 1585-1604 11 5'-CCG TCG TAG TAG TAA CAG ACT 138-160
TT-3' 12 5'-TGT CCA TAA TAG TAA TTT CCA A-3' 166-187 13 5'-CAG CAA
ATT ATG AGT CAT GCG GAT 211-236 TC-3' (**) target reference
numbering is in accordance with HDAC-1, GenBank Accession Number
U50079.
TABLE-US-00003 TABLE 2 SEQ ID TARGET NO. SEQUENCE (***) 14 5'-CTC
CTT GAC TGT ACG CCA TG-3' 1-20 15 5'-TGC TGC TGC TGC TGC TGC CG-3'
121-141 16 5'-CCT CCT GCT GCT GCT GCT GC-3' 132-152 17 5'-CCG TCG
TAG TAG TAG CAG ACT TT- 138-160 3' 18 5'-TGT CCA TAA TAA TAA TTT
CCA A-3' 166-187 19 5'-CAG CAA GTT ATG GGT CAT GCG GAT 211-236
TC-3' 20 5'-GGT TCC TTT GGT ATC TGT TT-3' 1605-1625 (***) target
reference numbering is in accordance with HDAC-2, GenBank Accession
Number U31814.
TABLE-US-00004 TABLE 3 SEQ ID TARGET NO. SEQUENCE (***) 21 5'-GCT
GCC TGC CGT GCC CAC CC-3' 514-533 (***) target reference numbering
is in accordance with HDAC-4
[0379] The following examples are intended to further illustrate
certain preferred embodiments of the invention, and are not
intended to limit the scope of the invention.
EXAMPLES
Preparation of Amines
Methyl-3-aminophenylacetate (1)
##STR00049##
[0381] To a solution of 3-aminophenylacetic acid (3 g, 19.85 mmol)
in methanol (50 mL) at room temperature was added HCl conc. (37%,
7.5 mL). The mixture was stirred 6 h at room temperature then
treated with a saturated aqueous solution of NaHCO.sub.3. The
solvent was removed under reduced pressure then the aqueous phase
was extracted several times with CH.sub.2Cl.sub.2. The combined
organic extracts were dried over (MgSO.sub.4) and evaporated. The
crude mixture was purified by flash chromatography using
hexane/AcOEt (1:1) yielding 1 as a yellow oil (3.06 g, 79%).
[0382] .sup.1H NMR: (300 MHz, CDCl.sub.3): .delta. 7.10 (t, J=8 Hz,
1H), 6.68-6.58 (m, 3H), 3.69-3.65 (m, 5H), 3.53 (s, 2H).
Methyl-4-aminophenyl benzoate (2)
##STR00050##
[0384] To a solution of 4-aminobenzoic acid (10 g, 72.92 mmol) in
methanol (200 mL) at room temperature was added HCl conc. (37%, 25
mL). The solution mixture was heated overnight at 70.degree. C.
Once the solution was clear (completed) the reaction was treated
with a saturated aqueous solution of NaHCO.sub.3 and
Na.sub.2CO.sub.3 powder until pH 9. The solvent was then evaporated
under reduced pressure and the aqueous phase was extracted several
times with AcOEt. The combined organic extracts were dried over
(MgSO.sub.4) and evaporated. The crude product 2 (9.30 g 85%) was
obtained as a beige solid and was clean enough to use without
further purification.
[0385] .sup.1H NMR: (300 MHz, CDCl.sub.3): .delta. 7.85 (d, J=8 Hz,
2H), 6.63 (d, J=8 Hz, 2H), 4.04 (broad s. 2H), 3.85 (s. 3H).
Methyl-4-aminophenylacetate (3)
##STR00051##
[0387] To a solution of 4-aminophenylacetic acid (10 g, 66 2 mmol)
in methanol (150 mL) at room temperature was added HCl conc. (37%
25 mL). The mixture became yellow and was stirred overnight. The
reaction mixture was then quenched with a saturated aqueous
solution of NaHCO.sub.3. The methanol was evaporated under reduced
pressure and the aqueous layer was extracted several times with
AcOEt. The combined organic extracts were dried over (MgSO.sub.4)
and evaporated. The crude residue was purified by flash
chromatography using hexane/AcOEt (4:1) as solvent mixture yielding
3 as a yellow oil (9.44 g, 74%).
[0388] .sup.1H NMR: (300 MHz, CDCl.sub.3): .delta. 7.05 (d, J=10
Hz, 2H), 6.65 (d, J=10 Hz, 2H), 3.65 (s, 3H), 3.63 (broad s, 2H),
3.51 (s, 2H).
Example 1
2-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-N-hydroxy-acetamide
(4)
##STR00052##
[0389] Step 1:
Methyl-2-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-acetate
(5)
##STR00053##
[0391] To a solution of 3 (500 mg, 2.56 mmol), in CH.sub.2Cl.sub.2
(8 mL) at room temperature were added Et.sub.3N (712 .mu..mu.L,
5.12 mmol) followed by 2-benzothiophenesulfonyl chloride (712 mg,
3.07 mmol). The mixture was stirred overnight at room temperature
then quenched with a saturated aqueous solution of NaHCO.sub.3. The
phases were separated and the aqueous layer was extracted several
times with CH.sub.2Cl.sub.2. The combined organic extracts were
dried over (MgSO.sub.4) and evaporated. The mixture of the mono and
bis alkylated products were dissolved in methanol (.about.8 mL) and
NaOMe was added (691 mg, 12.8 mmol). The resulting mixture was
heated at 60.degree. C. for 30 min the HCl 1N was added until pH 2.
Then a saturated aqueous solution of NaHCO.sub.3 was added until pH
7-8. The solvent was evaporated under reduced pressure then the
aqueous layer was extracted several times with CH.sub.2Cl.sub.2.
The combined organic extracts were dried over (MgSO.sub.4) and
evaporated. The residue was purified by flash chromatography using
toluene/AcOEt 7:3 as solvent mixture and a second flash
chromatography using CH.sub.2Cl.sub.2/acetone 98:2 as solvent
yielding the title compound 5 as yellowish powder (487 mg,
53%).
[0392] .sup.1H NMR: (300 MHz, CDCl.sub.3): .delta. 7.80 (d, J=8 Hz,
2H), 7.75 (s, 1H), 7.44 (m, 2H), 7.14 (m, 4H), 6.79 (broad s, 1H)
3.67 (s, 3H), 3.56 (s, 2H)
##STR00054##
Step 2: 2-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-acetic acid
(6)
[0393] To a solution of 5 from step 1 (451 mg, 1.25 mmol) in a
solvent mixture of THF (20 mL) and H.sub.2O (20 mL) at room
temperature was added LiOH (524 mg, 12.5 mmol). The mixture was
stirred for 2 h at room temperature and then was treated with a
saturated aqueous solution of NH.sub.4Cl. The resulting solution
was extracted several times with AcOEt. The combined organic
extracts were dried over (MgSO.sub.4). The crude residue was then
purified by flash chromatography using CH.sub.2Cl.sub.2/MeOH (9:1)
as solvent mixture yielding the title compound 6 as white solid
(404 mg, 93%).
[0394] .sup.1H NMR: (300 MHz, DMSO-d.sub.6): .delta. 8.03 (d, J=8
Hz, 1H), 7.97 (d, J=7 Hz, 1H), 7.92 (s, 1H), 7.50-7.45 (m, 2H),
7.13-7.06 (m, 4H), 3.44 (s, 2H).
Step 3:
2-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-N-hydroxy-acetamid-
e (4)
##STR00055##
[0395] Method A:
[0396] To a solution of 6 (150 mg, 0.432 mmol) in a solvent mixture
of CH.sub.2Cl.sub.2 (10 mL) and THF (5 mL) was added at room
temperature 1,3-dicyclohexylcarbodiimide (DCC, 116 mg, 0.563 mmol).
The reaction mixture was stirred 30 min at room temperature then
NH.sub.2OTHP (76 mg, 0.650 mmol) and dimethylaminopyridine (DMAP, 5
mg) were added. The solution was stirred over night at room
temperature and the solvents were evaporated under reduced
pressure. The crude material was purified by flash chromatography
using CH.sub.2Cl.sub.2/MeOH (9:1) as solvent. The residue was
dissolved in MeOH (-10 mL) and 10-camphorsulfonic acid (CSA, 100
mg, 0.432 mmol) was added. The mixture was stirred at room
temperature overnight then treated with a saturated aqueous
solution of NaHCO.sub.3. The solvent was evaporated under reduced
pressure and the aqueous phase was extracted several times with
CH.sub.2Cl.sub.2 (3.times.) and AcOEt (3.times.). The combined
organic extracts were dried over (MgSO.sub.4) and evaporated. The
crude product was purified by preparative high pressure liquid
chromatography on reversed phase silica gel using a gradient of
water/CH.sub.3CN (10-65%) yielding the title compound 4 as
yellowish solid (70 mg, 45%).
[0397] .sup.1H NMR (300 MHz, CD.sub.3OD): .delta. 7.92-7.88 (m,
2H), 7.80 (s, 1H), 7.50-7.45 (m, 2H), 7.23-7.16 (m, 4H) 3.35 (s,
2H).
[0398] Except where otherwise indicated, the following compounds
were prepared by procedures analogous to those described in Example
1, but substituting the sulfonyl chloride indicated for
2-benzothiophenesulfonyl chloride in step 1.
Example 2
2-[4-(2-nitrobenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide
(7)
##STR00056##
[0399] Sulfonyl chloride: 2-nitrobenzenesulfonyl chloride
[0400] Yield: Step 1: 82%
[0401] Yield: Step 2: 99%
[0402] Yield: Step 3: 19%
[0403] .sup.1H NMR (300 MHz, DMSO-d.sub.6); .delta. 10.59 (s, 1H);
8.78 (s, 1H); 7.94 (s, 2H), 7.81 (s, 2H), 7.20-7.02 (m, 4H); 3.13
(s, 2H).
Example 3
2-[4-(2.5-dichlorobenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide
(8)
##STR00057##
[0404] Sulfonyl chloride: 2.5-Dichlorobenzenesulfonyl chloride
[0405] Yield: Step 1: 66%
[0406] Yield: Step 2: 96%
[0407] Yield: Step 3: 66%
[0408] .sup.1H NMR (300 MHz, DMSO-d.sub.6); .delta. 10.68 (s, 1H),
8.88 (s, 1H), 7.95 (s, 1H), 7.67 (s, 2H); 7.13 (d, 2H, J=8 Hz 7.02
(d, 2H, J=8 Hz 3.16 (s, 2H)
Example 4
2-[4-(4-methylbenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide
(9)
##STR00058##
[0409] Sulfonyl chloride: 4-methylbenzenesulfonyl chloride
Step 1: Yield 100%
Step 2:
2-[4-(4-methylbenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide
(9)
Method B:
[0410] To a solution of
methyl-2-[4-(4-methylbenzenesulfonylamino)]phenylacetate (459 mg,
1.44 mmol) in methanol (10 mL), at room temperature were added
hydroxylamine hydrochloride (200 mg, 2.88 mmol) followed by sodium
methoxide (389 mg, 7.19 mmol). The resulting mixture was heated
overnight at 60.degree. C. then treated with HCl (1N) until pH 2.
The solvent was evaporated under reduced pressure then the aqueous
phase was extracted several times with CH.sub.2Cl.sub.2. The
combined organic extracts were dried over (MgSO.sub.4) then
evaporated. The crude mixture was purified by flash chromatography
using CH.sub.2Cl.sub.2/MeOH (9:1) as solvent mixture yielding the
title compound 9 (244 mg, 53%) as a white powder.
[0411] .sup.1H NMR (300 MHz, acetone-d.sub.6); .delta. 7.68 (d, J=8
Hz, 2H); 7.29 (d, J=8 Hz, 2H), 7.15 (br. s, 4H), 3.33 (s, 2H,
CH.sub.2), 2.33 (s, 3H, CH.sub.3).
[0412] The following compounds were prepared following procedures
analogous to those described in Example 1, step 1, and Example 4,
step 2 (Method B), but substituting the sulfonyl chloride indicated
for 2-benzothiophenesulfonyl chloride in step 1.
Example 5
2-[4-(3-trifluoromethylbenzenesulfonylamino)-phenyl]-N-hydroxy
acetamide (10)
##STR00059##
[0413] Sulfonyl chloride: 3-trifluoromethylbenzenesulfonyl
chloride
[0414] Yield: Step 1: 70%
[0415] Yield: Step 2: 49%
[0416] .sup.1H NMR (300 MHz, acetone-d.sub.6); .delta.=8.09 (s,
1H), 8.05 (d, 1H, J=8 Hz), 7.95 (d, 1H, J=8 Hz); 7.77 (t, 1H, J=8
Hz); 7.21 (d, 2H, J=8 Hz), 7.13 (d, 2H, J=8 Hz); 3.35 (s, 2H,
CH.sub.2)
Example 6
2-[4-(tert-butylsulfonylamino)-phenyl]-N-hydroxy-acetamide (11)
##STR00060##
[0417] Sulfonyl chloride: 4-tert-butylsulfonyl chloride
[0418] Yield: Step 1: 76%
[0419] Yield: Step 2: 40%
[0420] .sup.1H NMR (300 MHz, acetone-d.sub.6); .delta. 7.75 (d, 2H,
J=9 Hz), 7.56 (d, 2H, J=9 Hz); 7.17 (s, 4H); 3.34 (s, 2H), 1.29 (s,
9H)
[0421] The following compound was prepared following procedures
analogous to those described in Example 1, steps 1-2, substituting
the sulfonyl chloride indicated for 2-benzothiophenesulfonyl
chloride in step 1, followed by hydroxamic acid formation using
Method C
Example 7
2-[2-(naphthylsulfonylamino)-phenyl]-N-hydroxy-acetamide (12)
##STR00061##
[0423] Sulfonyl chloride: 2-naphthylsulfonyl chloride
[0424] Yield: Step 1: 100%
[0425] Yield: Step 2: 100%
Step 3: 2-[2-(naphthylsulfonylamino)-phenyl]-N-hydroxy-acetamide
(12)
Method C:
[0426] To a solution of 2-[2-(naphthylsulfonylamino)]-phenylacetic
acid (191 mg, 0.563 mmol) in CH.sub.2Cl.sub.2 (20 mL) at room
temperature were added DMF (5 drop) followed by (COCl).sub.2 (250
.mu.L, 2.81 mmol). The mixture became yellow and solidification
appeared. The reaction was stirred 90 min at room temperature then
(COCl).sub.2 was added until no bubbling (.about.1 mL). Then the
solvents were evaporated under reduced pressure. The crude material
was dissolved in CH.sub.2Cl.sub.2 and TMSONH.sub.2 (3 mL) was added
to the solution. The reaction was exothermic and the resulting
mixture was stirred 2 h at room temperature then treated with HCl
(1N) until pH 2. The phases were separated and the aqueous layer
was extracted several times with CH.sub.2Cl.sub.2. The combined
organic extracts were dried over (MgSO.sub.4) then evaporated. The
crude compound was purified 3 times by flash chromatography using
CH.sub.2Cl.sub.2/MeOH (9:1) as solvent mixture then another
purification using preparative high pressure liquid chromatography
using reversed phase chromatography with a gradient of
water/CH.sub.3CN (10-70%) yielding the title compound 12 as a white
powder (29 mg, 15%).
[0427] .sup.1H NMR (300 MHz, acetone-d.sub.6); .delta. 9.13 (s,
1H), 8.42 (s, 1H), 8.08-7.97 (m, 3H), 7.82 (dd, 1H, J=9 Hz, 1.5
Hz), 7.70-7.63 (m, 2H), 7.21-7.14 (m, 4H), 3.50 (s, 2H)
[0428] The following compound was prepared following procedures
analogous to those described in Example 1, steps 1-2, substituting
the indicated sulfonyl chloride and amine indicated for
2-benzothiophenesulfonyl chloride and 3 in step 1, followed by
hydroxamic acid formation using Method D.
Example 8
N-hydroxy-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-benzamide
(13)
##STR00062##
[0429] Sulfonyl chloride: 2-Benzothiophenesulfonyl chloride
[0430] Amine: Methyl-4-aminobenzoate (2)
[0431] Yield: Step 1: 80%
[0432] Yield: Step 2: 69%
Step 3:
N-hydroxy-[4-benzo[b]thiophene-2-sulfonylamino)-phenyl]-benzamide
(13)
Method D:
[0433] To a solution of
2-[4-benzo[b]thiophene-2-sulfonylamino]benzoic acid (300 mg, 0.90
mmol) in DMF (20 mL) at room temperature were added
1-(3-dimethyl-aminopropyl)-3-ethylcarbodiimide hydrochloride (EDC,
207 mg, 1.08 mmol), and 1-Hydroxybenzotriazole hydrate (HOBT, 182
mg, 1.35 mmol). The mixture was stirred 20 min. at room temperature
then NH.sub.2OTHP (158 mg, 1.35 mmol) was added. The resulting
mixture was heated at 50.degree. C. for 24 h then stirred at room
temperature for 24 h. The DMF solvent was evaporated under reduced
pressure and the residue was dissolved in CH.sub.2Cl.sub.2 and
washed with brine or a saturated aqueous solution of NaHCO.sub.3.
The combined organic extracts were dried over (MgSO.sub.4) then
condensed. The crude compound was purified by flash chromatography
using CH.sub.2Cl.sub.2/MeOH (9:1) as solvent mixture. The residue
was then dissolved in methanol (20 mL) then 10-camphorsulfonic acid
(CSA, 100 mg, 0.45 mmol) was added. The mixture was stirred 2 h at
room temperature then the solvents were evaporated under reduced
pressure at room temperature to avoid thermal decomposition. The
crude was purified by flash chromatography using
CH.sub.2Cl.sub.2/MeOH (9:1) as solvent mixture. A second
purification was performed using a preparative high pressure liquid
chromatography using a gradient of water/CH.sub.3CN (10-85%) as
solvent giving the title compound 13 as a red solid (212 mg,
68%).
[0434] .sup.1H NMR (300 MHz, acetone-d.sub.6); .delta. 10.69 (s,
1H), 9.70 (s, 1H); 8.01-7.97 (m, 3H), 7.77 (d, 2H, J=9 Hz);
7.55-7.39 (m, 4H).
Example 9
2-[3-benzo[b]thiopene-2-sulfonylamino)-phenyl]N-hydroxy-acetamide
(14)
##STR00063##
[0435] Sulfonyl chloride: 2-Benzothiophenesulfonyl chloride
[0436] Amine: Methyl-3-aminophenyl acetate (1)
[0437] Yield: Step 1: 88%
[0438] Yield: Step 2: 89%
[0439] Yield: Step 3: 32%
[0440] .sup.1H NMR (300 MHz, Acetoned.sub.6); .delta. 10.20 (s,
1H), 8.33 (s, 1H), 7.99-7.95 (m, 3H), 7.53-7.43 (m, 2H), 7.35 (s,
1H), 7.21-7.17 (m, 2H), 7.06-7.03 (m, 1H), 3.38 (s, 2H)
Example 10
2-[4-(3,4-dichlorobenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide
(15)
##STR00064##
[0441] Sulfonyl chloride: 3.4-Dichlorobenzenesulfonyl chloride
[0442] Yield: Step 1: 80%
[0443] Yield: Step 2: 67%
[0444] Yield: Step 3: 81%
[0445] .sup.1H NMR (300 MHz, acetone-d.sub.6); .delta. 10.12 (s,
1H), 9.15 (s, 1H), 7.92 (s, 1H), 7.74-7.71 (m, 2H), 7.23 (d, 2H,
J=9 Hz), 7.14 (d, 2H, J=9 Hz), 3.36 (s, 2H)
Example 11
2-[4-(2-Thiophenesulfonylamino)-phenyl]-N-hydroxy-acetamide
(16)
##STR00065##
[0446] Sulfonyl chloride: 2-Thiophenesulfonyl chloride
[0447] Yield: Step 1: 84%
[0448] Yield: Step 2: 83%
[0449] Yield: Step 3: 9%
[0450] .sup.1H NMR (300 MHz, acetone-d.sub.6); .delta. 7.78 (s,
1H), 7.53 (s, 1H), 7.21 (s, 4H), 7.09 (s, 1H), 3.37 (s, 2H)
Example 12
2-[4-(3-nitrobenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide
(17)
##STR00066##
[0451] Sulfonyl chloride: 3-Nitrobenzenesulfonyl chloride
[0452] Yield: Step 1: 47%
[0453] Yield: Step 2: 34%
[0454] Yield: Step 3: 16%
[0455] .sup.1H NMR (300 MHz, acetone-d.sub.6); .delta. 9.31 (s,
1H), 8.59 (s, 1H), 8.45 (d, 1H, J=8 Hz), 8.16 (d, 1H, J=8 Hz), 7.85
(t, 1H, J=8 Hz), 7.20-7.14 (m, 4H), 3.35 (s, 2H)
Example 13
2-[4-(8-quinolinesulfonylamino)-phenyl]-N-hydroxy-acetamide
(18)
##STR00067##
[0456] Sulfonyl chloride: 8-quinolinesulfonyl chloride
[0457] Yield: Step 1: 83%
[0458] Yield: Step 2: 78%
[0459] Yield: Step 3: 42%
[0460] .sup.1H NMR (300 MHz, acetone-d.sub.6); .delta. 9.17 (s,
1H), 8.50 (d, 1H, J=8 Hz), 8.33 (d, 1H, J=8 Hz), 8.21 (d, 1H, J=8
Hz), 7.71-7.68 (m, 3H), 7.05 (broad s., 4H), 3.22 (s, 2H)
Example 14
2-[4-(4-bromobenzenesulfonylamino)-phenyl]-N-hydroxy-acetamide
(19)
##STR00068##
[0461] Sulfonyl chloride: 4-Bromobenzenesulfonyl chloride
[0462] Yield: Step 1: 80%
[0463] Yield: Step 2: 81%
[0464] Yield: Step 3: 48%
[0465] .sup.1H NMR (300 MHz, acetone-d.sub.6); .delta. 9.17 (s,
1H), 7.72 (s, 4H), 7.19-7.14 (m, 4H), 3.35 (s, 2H)
Example 15
N-Hydroxy-5-[3-benzenesulfonylamino)-phenyl]-pentanamide (26)
##STR00069##
[0466] Step 1: 3-(benzenesulfonylamino)-phenyl iodide (21)
[0467] To a solution of 3-iodoaniline (5 g, 22 8 mmol), in
CH.sub.2Cl.sub.2 (100 mL), were added at room temperature Et.sub.3N
(6.97 mL) followed by benzenesulfonyl chloride (5.84 mL). The
mixture was stirred 4 h then a white precipitate was formed. A
saturated aqueous solution of NaHCO.sub.3 was added and the phases
were separated. The aqueous layer was extracted several times with
CH.sub.2Cl.sub.2 and the combined extracts were dried over
(MgSO.sub.4) then evaporated. The crude mixture was dissolved in
MeOH (100 mL) and NaOMe (6 g), was added and the mixture was heated
1 h at 60.degree. C. The solution became clear with time and HCl
(1N) was added. The solvent was evaporated under reduced pressure
then the aqueous phase was extracted several times with
CH.sub.2Cl.sub.2. The combined organic extracts were dried over
(MgSO.sub.4) and evaporated. The crude material was purified by
flash chromatography using (100% CH.sub.2Cl.sub.2) as solvent
yielding the title compound 21 (7.68 g, 94%) as yellow solid.
[0468] .sup.1H NMR: (300 MHz, CDCl.sub.3): .delta. 7.82-7.78 (m,
2H), 7.60-7.55 (m, 1H), 7.50-7.42 (m, 4H), 7.10-7.06 (m, 1H), 6.96
(t, J=8 Hz, 1H), 6.87 (broad s, 1H).
Step 2: 3-(benzenesulfonylamino)-phenyl-propargylic alcohol
(22)
[0469] To a solution of 21 (500 mg, 1.39 mmol) in pyrrolidine (5
mL) at room temperature was added Pd(PPh.sub.3).sub.4 (80 mg, 0.069
mmol), followed by CuI (26 mg, 0.139 mmol). The mixture was stirred
until complete dissolution. Propargylic alcohol (162 .mu.L, 2.78
mmol) was added and stirred 6 h at room temperature. Then the
solution was treated with a saturated aqueous solution of
NH.sub.4Cl and extracted several times with AcOEt. The combined
organic extracts were dried over (MgSO.sub.4) then evaporated. The
residue was purified by flash chromatography using hexane/AcOEt
(1:1) as solvent mixture yielding 22 (395 mg, 99%) as yellow
solid.
[0470] .sup.1H NMR: (300 MHz, CDCl.sub.3): .delta. 7.79-7.76 (m,
2H), 7.55-7.52 (m, 1H), 7.45 (t, J=8 Hz, 2H), 7.19-7.15 (m, 3H),
7.07-7.03 (m, 1H), 4.47 (s, 2H).
Step 3: 5-[3-(benzenesulfonylamino)-phenyl]-4-yn-2-pentenoate
(23)
[0471] To a solution of 22 (2.75 g, 9.58 mmol) in CH.sub.3CN (150
mL) at room temperature were added 4-methylmorpholine N-oxide (NMO,
1.68 g, 14.37 mmol) followed by tetrapropylammonium perruthenate
(TPAP, 336 mg, 0.958 mmol). The mixture was stirred at room
temperature 3 h, and then filtrated through a Celite pad with a
fritted glass funnel. To the filtrate
carbethoxymethylenetriphenyl-phosphorane (6.66 g, 19.16 mmol) was
added and the resulting solution was stirred 3 h at room
temperature. The solvent was evaporated and the residue was
dissolved in CH.sub.2Cl.sub.2 and washed with a saturated aqueous
solution of NH.sub.4Cl. The aqueous layer was extracted several
times with CH.sub.2Cl.sub.2 then the combined organic extract were
dried over (MgSO.sub.4) and evaporated. The crude material was
purified by flash chromatography using hexane/AcOEt (1:1) as
solvent mixture giving 23 (1.21 g, 36%) as yellow oil.
[0472] .sup.1H NMR: (300 MHz, CDCl.sub.3): .delta. 7.81 (d, J=8 Hz,
2H), 7.56-7.43 (m, 3H), 7.26-7.21 (m, 3H), 7.13-7.11 (m, 1H), 6.93
(d, J=16 Hz, 1H), 6.29 (d, J=16 Hz, 1H), 4.24 (q, J=7 Hz, 2H), 1.31
(t, J=7 Hz, 3H).
Step 4: 5-[3-(benzenesulfonylamino)-phenyl]-4-yn-2-pentenic acid
(24)
[0473] To a solution of 23 (888 mg, 2.50 mmol) in a solvent mixture
of THF (10 mL) and water (10 mL) at room temperature was added LiOH
(1.04 g, 25.01 mmol). The resulting mixture was heated 2 h at
60.degree. C. and treated with HCl (1N) until pH 2. The phases were
separated and the aqueous layer was extracted several times with
AcOEt. The combined organic extracts were dried over (MgSO.sub.4)
then evaporated. The crude residue was purified by flash
chromatography using CH.sub.2Cl.sub.2/MeOH (9:1) as solvent mixture
yielding 24 (712 mg, 88%), as white solid.
[0474] .sup.1H NMR: (300 MHz, DMSO-d.sub.6): .delta. 7.78-7.76 (m,
2H), 7.75-7.53 (m, 3H), 7.33-7.27 (m, 1H), 7.19-7.16 (m, 3H), 6.89
(d, J=16 Hz, 1H), 6.33 (d, J=16 Hz, 1H).
Step 5: 5-[3-(benzenesulfonylamino)-phenyl]-pentanoic acid (25)
[0475] To a solution 24 (100 mg, 0.306 mmol), in MeOH (6 mL) at
room temperature was added a solution of Pd/C (10%, 20 mg, 1 mL
MeOH). The reaction mixture was degassed and purged several times
with H.sub.2 gas with a final pressure of 60 psi. The mixture was
stirred 2 h at room temperature then the resulting solution was
filtrated over a silica gel pad with a fritted glass funnel. The
solvent was evaporated yielding 25 (68 mg, 96%) and it was used
directly for the next step without further purification.
[0476] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta. 7.81-7.78
(m, 2H), 7.56-7.46 (m, 3H), 7.11-7.01 (m, 3H), 6.87 (d, J=8 Hz,
1H), 2.49 (broad s, 2H), 2.25 (broad s, 2H), 1.52 (broad s, 4H)
Step 6: N-Hydroxy-5-[3-benzenesulfonylamino)-phenyl]-pentanamide
(26)
[0477] To a solution of 25 (100 mg, 300 mmol) in DMF (10 mL) at
room temperature were added
1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (EDC,
69 mg, 0.320 mmol), and 1-hydroxybenzotriazole hydrate (HOBT, 61
mg, 0.45 mmol). The mixture was stirred 20 min. at room temperature
then NH.sub.2OTHP (53 mg, 0.45 mmol) was added. The resulting
mixture was heated overnight at 50.degree. C. The DMF solvent was
evaporated under reduced pressure and the residue was dissolved in
CH.sub.2Cl.sub.2 and washed with brine or a saturated aqueous
solution of NaHCO.sub.3. The combined organic extracts were dried
over (MgSO.sub.4) then evaporated. The crude compound was purified
by flash chromatography using hexane/acetone (7:3) as solvent
mixture. The residue was then dissolved in MeOH (20 mL) then
10-camphorsulfonic acid (CSA, 35 mg, 150 mmol) was added. The
mixture was stirred 2 h at room temperature then the solvents were
evaporated under reduced pressure at room temperature to avoid
thermal decomposition. The crude mixture was purified by flash
chromatography using CH.sub.2Cl.sub.2/MeOH (9:1) as solvent mixture
giving 26 as a yellowish solid (62 mg, 60%).
[0478] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta.=7.80-7.78
(m, 2H), 7.56-7.52 (m, 3H), 7.13-6.89 (m, 4H), 2.52 (broad s, 2H),
2.10 (broad s, 2H), 1.53 (broad s, 4H)
Example 16
N-Hydroxy-5-[4-(benzenesulfonylamino)-phenyl]-4-yn-2-pentanamide
(32)
##STR00070##
[0479] Step 1: 4-(benzenesulfonylamino)-phenyl iodide (28)
[0480] Compound 28 was prepared using the procedure described in
Example 15, step 1, but substituting 4-iodoaniline for
3-iodoaniline.
[0481] Yield: 97%
[0482] .sup.1H NMR: (300 MHz, CDCl.sub.3): .delta. 9.15 (broad s,
1H), 7.82 (d, J=8 Hz, 2H), 7.68-7.51 (m, 5H), 7.05 (d, J=8 Hz,
2H).
Step 2: 4-(benzenesulfonylamino)-phenyl-propargylic alcohol
(29)
[0483] Compound 29 was prepared using the procedure described in
Example 15, step 2 but substituting compound 21 for compound
28.
[0484] Yield: 61%
[0485] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta. 7.83-7.80
(m, 2H), 7.62-7.51 (m, 3H), 7.30 (d, J=8 Hz, 2H), 7.21 (d, J=8 Hz,
2H), 4.36 (s, 2H), 2.80 (broad s, 2H).
Step 3: 5-[4-(benzenesulfonylamino)-phenyl]-4-yn-2-pentenoate
(30)
[0486] Compound 30 was prepared using the procedure described in
Example 15, step 3 but substituting compound 22 for compound 29
[0487] Yield: 16%
[0488] .sup.1H NMR: (300 MHz, CDCl.sub.3): .delta. 7.81-7.78 (m,
2H), 7.59-7.43 (m, 3H), 7.34 (d, J=8 Hz, 2H), 7.05 (d, J=8 Hz, 2H),
6.93 (d, J=16 Hz, 1H), 6.26 (d, J=16 Hz, 1H), 4.23 (q, J=7 Hz, 2H),
1.30 (t, J=7 Hz, 3H).
Step 4: 5-[4-(benzenesulfonylamino)-phenyl]-4-yn-2-pentenic acid
(31)
[0489] Compound 31 was prepared using the procedure described in
Example 15 step 4 but substituting compound 23 for compound 30
[0490] Yield: 92%
[0491] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta. 7.87-7.84
(m, 2H), 7.62 (m, 3H), 7.42 (d, J=8 Hz, 2H), 7.28 (d, J=8 Hz, 2H),
6.94 (d, J=16 Hz, 1H), 6.29 (d, J=16 Hz, 1H).
Step 5:
N-hydroxy-5-[4-(benzenesulfonylamino)-phenyl]-4-yn-2-pentanamide
(32)
[0492] Compound 32 was prepared using the procedure described in
Example 15 step 6 but substituting compound 25 for compound 31
[0493] Yield: 78%
[0494] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta. 7.84 (broad
s, 2H), 7.60-7.55 (m, 3H), 7.38-7.30 (m, 4H), 6.84 (d, J=16 Hz,
1H), 6.40 (d, J=16 Hz, 1H).
Example 17
N-Hydroxy-5-[4-benzenesulfonylamino)-phenyl]-pentanamide (34)
Step 1: 5-[4-(benzenesulfonylamino)-phenyl]-pentanoic acid (33)
[0495] Compound 33 was prepared using the procedure described in
Example 15 step 5 but substituting compound 24 for compound 31.
[0496] Yield: 100%
[0497] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta.=7.78-7.75
(m, 2H), 7.56-7.46 (m, 3H), 7.16-7.05 (m, 4H), 2.52 (broad s, 2H),
2.29-2.25 (m, 2H), 1.56 (broad s, 4H).
Step 2: N-Hydroxy-5-[4-benzenesulfonylamino)-phenyl]-pentanamide
(34)
[0498] Compound 34 was prepared using the procedure described in
Example 15 step 6 but substituting compound 25 for compound 33.
[0499] Yield: 62%
[0500] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta. 7.78-7.75
(m, 2H), 7.59-7.51 (m, 3H), 7.09 (broad s, 4H), 2.85 (broad s, 1H),
2.53 (broad s, 2H), 2.05 (broad s, 2H), 1.56 (broad s, 4H).
Example 18
N-Hydroxy-3-[4-(benzenesulfonylamino)-phenyl]-2-propenamide
(36)
##STR00071##
[0501] Step 1: 3-[4-(benzenesulfonylamino)-phenyl]-2-propenoic acid
(35)
[0502] To a solution of 28 (500 mg, 1.39 mmol), in DMF (10 mL) at
room temperature were added
tris(dibenzylideneacetone)dipalladium(0) (Pd.sub.2(dba).sub.3; 38
mg, 1.67 mmol), tri-o-tolylphosphine (P(o-tol).sub.3, 25 mg, 0.83
mmol), Et.sub.3N (483 .mu..mu.L, 3.48 mmol) and finally acrylic
acid (84 .mu..mu.L, 1.67 mmol). The resulting solution was degassed
and purged several times with N.sub.2 then heated overnight at
100.degree. C. The solution was filtrated through a Celite pad with
a fritted glass funnel then the filtrate was evaporated. The
residue was purified by flash chromatography using
CH.sub.2Cl.sub.2/MeOH (95:5) as solvent mixture yielding the title
compound 35 (415 mg, 99%) as yellowish solid.
[0503] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta. 7.88-7.85
(m, 2H), 7.62-7.55 (m, 6H), 7.29 (d, J=9 Hz, 2H), 6.41 (d, J=16 Hz,
1H), 2.95 (s, 1H), 2.79 (s, 1H).
Step 2: N-Hydroxy-3-[4-(benzenesulfonylamino)-phenyl]-2-propenamide
(36)
[0504] To a solution of 35 (200 mg, 0.660 mmol) in DMF (10 mL) at
room temperature were added
1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (EDCI,
151 mg, 0.79 mmol), and 1-Hydroxybenzotriazole hydrate (HOBT, 134
mg, 0.99 mmol). The mixture was stirred 20 min. at room temperature
then NH.sub.2OTHP (116 mg, 0.99 mmol) was added. The resulting
mixture was heated at 50.degree. C. for 24 h then the DMF solvent
was evaporated under reduced pressure and the residue was dissolved
in CH.sub.2Cl.sub.2, washed with a saturated aqueous solution of
NaHCO.sub.3. The combined organic extracts were dried over
(MgSO.sub.4) then condensed. The crude compound was purified by
flash chromatography using Hexane/acetone (7:3) as solvent mixture.
The residue was then dissolved in MeOH (10 mL) then
10-camphorsulfonic acid (CSA, 77 mg, 0.33 mmol) was added. The
mixture was stirred 2 h at room temperature then the solvents were
evaporated under reduced pressure at room temperature to avoid
thermal decomposition. The crude product was purified by flash
chromatography using CH.sub.2Cl.sub.2/MeOH (9:1) as solvent mixture
giving compound 36 (116 mg, 55%) as a orange solid.
[0505] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta. 7.85-7.83
(m, 2H), 7.64-7.47 (m, 6H), 7.26 (d, J=8 Hz, 2H), 6.48 (m, 1H),
2.82 (s, 1H), 2.79 (s, 1H).
Example 19
N-Hydroxy-3-[4-(benzenesulfonylamino)-phenyl]-2-propanamide
(38)
Step 1: 3-[4-(benzenesulfonylamino)-phenyl]-2-propionic acid
(37)
[0506] To a solution of 35 (350 mg, 1.16 mmol) in MeOH (15 mL) at
room temperature was added a solution of Pd/C 10% (50 mg. in MeOH
.about.3 mL). Then the resulting solution was purged several times
with H.sub.2 with a final pressure of 60 psi. The solution was
stirred 4 h then filtrated through a Celite pad with a fritted
glass funnel. The filtrate was evaporated and the residue compound
37 was pure enough to use for the next step without further
purification.
[0507] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta. 8.92 (broad
s, 1H), 7.79-7.76 (m, 2H), 7.60-7.47 (m, 3H), 7.12 (s, 4H), 3.32
(s, 1H), 2.81 (t, J=8 Hz, 2H), 2.53 (t, J=8 Hz, 2H).
Step 2: N-Hydroxy-3-[4-(benzenesulfonylamino)-phenyl]-2-propanamide
(38)
[0508] To a solution of 37 (1.16 mmol) in DMF (10 mL) at room
temperature were added
1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (EDC,
266 mg, 1.39 mmol), and 1-Hydroxybenzotriazole hydrate (HOBT, 235
mg, 1.74 mmol). The mixture was stirred 20 min. at room temperature
then NH.sub.2OTHP (204 mg, 1.74 mmol) was added. The resulting
mixture was heated at 50.degree. C. for 24 h then the DMF solvent
was condensed under reduced pressure and the residue was dissolved
in CH.sub.2Cl.sub.2, washed with a saturated aqueous solution of
NaHCO.sub.3. The combined organic extracts were dried over
(MgSO.sub.4) then evaporated. The crude compound was purified by
flash chromatography using Hexane/acetone (7:3) as solvent mixture.
The residue was then dissolved in MeOH (10 mL) then
10-camphorsulfonic acid (CSA, 135 mg, 0.58 mmol) was added. The
mixture was stirred 2 h at room temperature then the solvents were
evaporated under reduced pressure at room temperature to avoid
thermal decomposition. The crude was purified by flash
chromatography using CH.sub.2Cl.sub.2/MeOH (9:1) as solvent mixture
giving the title compound 38 (237 mg, 64%, for the last 3 steps) as
a yellow solid.
[0509] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta. 8.91 (broad
s, 1H), 7.78-7.76 (m, 2H), 7.57-7.51 (m, 3H), 7.10 (broad s, 4H),
2.82 (broad s, 2H), 2.34 (broad s, 2H), 1.07 (s, 1H), 0.85 (s,
1H).
Example 20
N-Hydroxy-4-[4-(benzenesulfonylamino)-phenyl]-butanamide (42)
##STR00072##
[0510] Step 1: Methyl-4-(4-aminophenyl)-butanoate (40)
[0511] To a solution of 4-(4-aminophenyl)-butyric acid (5 g, 27.90
mmol) in MeOH (100 mL) at room temperature was added HCl conc. (37%
15 mL). The resulting mixture was stirred overnight at 50.degree.
C. then treated with a saturated aqueous solution NaHCO.sub.3 and
Na.sub.2CO.sub.3 solid until pH 9. The solvent was evaporated under
reduced pressure then the aqueous phase was extracted several times
with CH.sub.2Cl.sub.2. The crude material was purified by flash
chromatography using CH.sub.2Cl.sub.2/MeOH as solvent mixture
yielding 40 (4.93 g, 91%) as orange solid.
[0512] .sup.1H NMR: (300 MHz, acetone-d.sub.6): .delta. 6.89 (d,
J=8 Hz, 2H), 6.59 (d, J=8 Hz, 2H), 4.40 (broad s, 1H), 3.60 (s,
3H), 2.48 (t, J=7 Hz, 2H), 2.28 (t, J=7 Hz, 2H), 1.82 (qt, J=7 Hz,
2H).
Step 2: 4-[4-(benzenesulfonylamino)-phenyl]-butyric acid (41)
[0513] To a solution of 40 (500 mg, 2.59 mmol) in CH.sub.2Cl.sub.2
at room temperature were added Et.sub.3N (901 .mu..mu.L, 6.48 mmol)
followed by benzenesulfonyl chloride (661 .mu.L, 5.18 mmol). The
mixture was stirred overnight at room temperature then treated with
a saturated aqueous solution of NH.sub.4Cl. The phases were
separated and the organic layer was extracted several times with
CH.sub.2Cl.sub.2. The combined organic extracts were dried over
(MgSO.sub.4) then evaporated under reduced pressure. The residue
was dissolved in a solvent mixture of THF (25 mL) and water (25 mL)
then LiOH (1.08 g, 25.9 mmol) was added. The mixture was heated at
50.degree. C. for 1 h then treated with HCl (1N) until pH2. The
phases were separated and the aqueous layer was extracted several
times with AcOEt. The combined organic extracts were dried over
(MgSO.sub.4) then evaporated. The crude was purified by flash
chromatography using CH.sub.2Cl.sub.2/MeOH (95:5) as solvent
mixture yielding 41 (800 mg, 96%) as a white solid
[0514] .sup.1H NMR: (300 MHz, CDCl.sub.3): .delta. 8.82 (1H, s
broad), 7.77-7.74 (2H, m), 7.55-50 (1H, m), 7.44-7.39 (2H, m),
7.05-6.97 (4H, m), 2.58 (2H, t, J=7 Hz), 2.31 (2H, t, J=7 Hz), 2.17
(1H, s), 1.94-1.84 (2H, m).
Step 3: N-Hydroxy-4-[4-(benzenesulfonylamino)-phenyl]-butanamide
(42)
[0515] To a solution 41 (800 mg, 2.59 mmol) in DMF (20 mL) at room
temperature were added
1-(3-Dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride (EDC,
593 mg, 3.12 mmol), and 1-Hydroxybenzotriazole hydrate (HOBT, 524
mg, 3.89 mmol). The mixture was stirred 20 min. at room temperature
then NH.sub.2OTHP (455 mg, 3.89 mmol) was added. The resulting
mixture was heated at 50.degree. C. for 24 h then the DMF solvent
was evaporated under reduced pressure and the residue was dissolved
in CH.sub.2Cl.sub.2, washed with a saturated aqueous solution of
NaHCO.sub.3. The combined organic extracts were dried over
(MgSO.sub.4) then evaporated. The crude compound was purified by
flash chromatography using Hexane/acetone (7:3) as solvent mixture.
The residue was then dissolved in MeOH (30 mL) then
10-camphorsulfonic acid (CSA, 300 mg, 1.30 mmol) was added. The
mixture was stirred 2 h at 50.degree. C. then the solvents were
condensed under reduced pressure at room temperature to avoid
thermal decomposition. The crude was purified by flash
chromatography using CH.sub.2Cl.sub.2/MeOH (9:1) as solvent mixture
giving the title compound 42 (115 mg, 13%) as a yellowish
solid.
[0516] .sup.1H NMR: (300 MHz, CDCl.sub.3): .delta. 7.79-7.76 (m,
2H), 7.61-7.48 (m, 3H), 7.13-7.05 (m, 4H), 2.83 (broad s, 1H), 2.53
(t, J=7 Hz, 2H), 2.14-2.04 (m, 2H), 1.83 (t, J=7 Hz, 2H).
Example 21
N-Hydroxy-4-(3-oxo-3-phenylpropenyl)-benzamide (45)
##STR00073##
[0517] Step 1: 4-(3-oxo-3-phenylpropenyl)-benzoic acid (43)
[0518] Sodium methoxide (1.8 g, 33 3 mmol) was added to a stirred
suspension of 4-carboxybenzaldehyde (2.5 g, 16.6 mmol) and
acetophenone (2.0 g uL, 16 6 mmol) in methanol (50 mL) at room
temperature. The mixture was stirred at room temperature for 16
hours, and half of the volume of methanol was removed under reduced
pressure. The mixture was poured into HCI 1M (50 mL) (until pH=2)
and ethyl acetate was added. The separated aqueous layer was
extracted with ethyl acetate (3.times.30 mL) dried (MgSO.sub.4
anh.), filtered and evaporated. The residue was triturated with
dichloromethane-hexanes (1:1) to afford 3 g of 43 (72% yield).
[0519] .sup.1H NMR (300 MHz, CDCl.sub.3); .delta. 7.50-7.87 (m,
7H), 8.04 (d, 2H, J=8 Hz), 8.16 (d, 2H, J=8 Hz)
Step 2:
4-(3-oxo-3-phenylpropenyl)-N--(O-tetrahydropyranyl)-benzamide
(44)
[0520] The carboxylic acid 43 (260 mg, 1.0 mmol) was dissolved in
anhydrous CH.sub.2Cl.sub.2 (10 mL) and DCC (256 mg, 1.2 mmol)
followed by NH.sub.2OTHP (145 mg, 1.2 mmol) were added. The mixture
was allowed to stir at room temperature for 2 h. Added NH.sub.4Cl
sat. and extracted with EtOAc. The organic layer was dried over
MgSO.sub.4, filtered and the solvent was evaporated under vacuum.
(Purification by column chromatography using 1%
MeOH/CH.sub.2Cl.sub.2 give the title compound which was used
directly in the next step.
Step 3: N-Hydroxy-4-(3-oxo-3-phenylpropenyl)-benzamide (45)
[0521] The protected hydroxamic acid 44 (234 mg, 0.67 mmol) was
dissolved in MeOH (7 mL) then CSA (31 mg, 0.13 mmol) was added. The
mixture was allowed to stir at reflux for 2 hours or until the
reaction was complete by TLC. Added HCl 1N, extracted with EtOAc,
dried the organic layer over anhydrous MgSO.sub.4 and filtered. The
solvent was evaporated under vacuum. Purification by column
chromatography using 5% MeOH/CH.sub.2Cl.sub.2, gave the title
compound.
[0522] .sup.1H NMR (300 MHz, DMSO-d.sub.6), .delta. 7.53-8.20 (m,
11H); 9.12 (br. s, 1H); 11.35 (br. s, 1H)
Example 22
N-Hydroxy-4-(3-oxo-3-phenylpropyl)-benzamide (50)
##STR00074##
[0523] Step 1: Methyl-4-(3-oxo-3-phenylpropenyl)-benzoate (46)
[0524] To 4-carbomethoxybenzaldehyde (79 mg, 0.48 mmol) and
acetophenone (56 .mu..mu.L, 0.48 mmol) in anhydrous methanol (1.6
mL), was added neat sodium methoxide (26 mg, 0.48 mmol). The
mixture was stirred at room temperature overnight then heated to
reflux for 1 hour, cooled down to room temperature and added HCl 1N
and EtOAc. The layers were separated and the organic layer dried
over anhydrous MgSO.sub.4 and filtered. The solvent was evaporated
under vacuum to afford a yellow solid, which was recrystallized
from acetonitrile/water to give a pale yellow crystalline
solid.
[0525] .sup.1H NMR (300 MHz, CDCl.sub.3); .delta. 3.95 (s, 3H),
7.50-8.12 (m, 11H)
Step 2: Methyl-4-(3-oxo-3-phenylpropyl)-benzoate (47)
[0526] The aromatic enone 46 (321 mg, 1.20 mmol) was dissolved in
anhydrous THF (6 mL) and anhydrous MeOH (6 ml). Added 2 small
scoops of Pd 10% on activated C, placed under an atmosphere of
hydrogen and allowed to stir for 2 hours at room temperature.
Purged with nitrogen, filtered through Celite and removed solvent
by evaporation under vacuum. The benzylic alcohol is reoxidized to
the ketone by the following procedure. The crude was taken back in
anhydrous CH.sub.2Cl.sub.2 (10 mL), with 3 .ANG. molecular sieves,
TPAP (1 scoop) was added followed by NMO (212 mg, 1.8 mmol).
Stirred at room temperature for 30 minutes and filtered through a
plug of silica gel. Solvent was evaporated under vacuum and
purified by column chromatography using 10% EtOAc/Hexane.
[0527] .sup.1H NMR (300 MHz, CDCl.sub.3); .delta. 3.14 (t, 2H),
3.34 (t, 2H), 3.90 (s, 3H), 7.30-7.60 (m, 6H), 7.92-7.99 (m,
4H).
Step 3: 4-(3-oxo-3-phenylpropyl)-benzoic acid (48)
[0528] To a solution of methyl ester 47 (195 mg, 0.73 mmol) in
water/THF (1:1, 0.07M) was added LiOH (46 mg, 1.1 mmol). The
resulting solution was stirred overnight at room temperature or
until no starting material was detected by TLC. HCl 1N was added
and the solution was extracted with EtOAc and the organic layer was
dried over anhydrous MgSO.sub.4. Filtration and evaporation of the
solvent under vacuum followed by purification by column
chromatography using 10% MeOH/CH.sub.2Cl.sub.2, gave the title
compound.
[0529] .sup.1H NMR (300 MHz, CDCl.sub.3); .delta. 3.16 (t, 2H),
3.36 (t, 2H), 7.33-7.60 (m, 5H), 7.93-8.06 (m, 4H).
Step 4: N-hydroxy-4-(3-oxo-3-phenylpropyl)-benzamide (50)
[0530] Following the procedure described in Example 21, Steps 2-3,
but substituting compound 48 for carboxylic acid 4, the title
compound was obtained.
[0531] .sup.1H NMR (300 MHz, DMSO-d.sub.6); .delta. 2.97 (t, 2H),
3.38 (t, 2H), 7.34 (d, 2H, J=8 Hz), 7.45-7.70 (m, 5H), 7.96 (dd,
2H, J=8 Hz, 1 Hz), 11.14 (br. s, 1H)
Example 23
N-Hydroxy-4-(3-oxo-3-phenyl-1-hydroxypropyl)-benzamide (53)
##STR00075##
[0532] Step 1: 4-Carboxy-N--(O-tetrahydropyranyl)-benzamide
(51)
[0533] Hydroxylamine-O-THP (3.9 g, 33 2 mmol) was added to a
suspension of 4-formylbenzoic acid (4.2 g, 27.7 mmol) and DCC (6.8
g, 33 2 mmol) in dichloromethane (200 mL). The mixture was stirred
at room temperature overnight and quenched with saturated ammonium
chloride. The separated aqueous layer was extracted with ethyl
acetate (3.times.100 ml) and the combined organic layers were
washed with brine, dried (MgSO.sub.4 anh), filtered and evaporated.
Flash chromatography of the residue (10% methanol in
CH.sub.2Cl.sub.2), afforded (51).
[0534] .sup.1H NMR (300 MHz, CDCl.sub.3); .delta. ppm. 10.04 (s,
1H), 8.95 (s, 1H), 7.99 (d, 2H, J=7.0 Hz), 7.93 (d, 2H, J=7.0 Hz),
5.1 (s, 1H), 3.60 (m, 2H), 1.60 (m, 6H)
Step 2:
4-(3-oxo-3-phenyl-1-hydroxypropyl)-N--(O-tetrahydropyranyl)-benzam-
ide (52)
[0535] n-BuLi (1.4M/hexane, 1.6 mL, 2.2 mmol) was added to a
0.degree. C. solution of diisopropylamine (337 .mu.L, 2.4 mmol) in
anhydrous THF (15 mL). Stirred at 0.degree. C. 10 minutes, then
cooled to -78.degree. C. Added acetophenone, then stirred 30
minutes at -78.degree. C. Cannulated into a -78.degree. C. solution
of the aldehyde 9 (50 mg, 2.0 mmol) in anhydrous THF (10 mL).
Stirred 3 hours at -78.degree. C., then added NH.sub.4Cl. Warmed to
room temperature, extracted with EtOAc, dried over MgSO.sub.4,
filtered and evaporated solvent under vacuum. Purification by HPLC
CH.sub.3CN: H.sub.2O: TFA 0.1%; 10-95% gave the title compound
52.
Step 3: N-Hydroxy-4-(3-oxo-3-phenyl-1-hydroxypropyl)-benzamide
(53)
[0536] Following the same procedure as described in Example 21,
Step 3, but substituting compound 52 for compound 44, the title
compound was obtained.
[0537] .sup.1H NMR (300 MHz, DMSO-d.sub.6); .delta. 3.20 (dd, 1H,
J=4 Hz, J=16 Hz), 3.42 (dd, 1H=16 Hz, 8 Hz), 5.20 (m, 1H),
7.44-8.18 (m, 9H), 11.15 (br. s, 1H), 11.32 (br. s, 1H)
Example 24
N-Hydroxy-4-(3-phenylpropyl)-benzamide (56)
##STR00076##
[0538] Step 1: 4-(3-phenylpropenyl)-benzoic
acid/4-(3-phenyl-2-propenyl)-benzoic acid (54)
[0539] Allylbenzene (255 .mu..mu.L, 1.9 mmol), 4-bromobenzoic acid
(523 mg, 2 6 mmol), Et.sub.3N (0.91 mL, 6.5 mmol), Palladium (II)
Acetate (16 mg, 0.052 mmol), triphenylphosphine (60 mg, 0.21 mmol)
and acetonitrile (5 mL) were stirred at reflux overnight in a round
bottom flask. Added HCl 1N, extracted with EtOAc, dried the organic
layer on anhydrous MgSO.sub.4, filtered, evaporated solvent under
vacuum. Purified by column chromatography using 10%
MeOH/CH.sub.2Cl.sub.2 yielded 90 mg (14%) of mixture of two
regioisomers 54. The mixture was then submitted for hydrogenation
without further characterization.
Step 2: 4-(3-phenylpropyl)-benzoic acid (55)
[0540] A mixture of regioisomeric olefins 54 (100 mg, 0.42 mmol)
and Pd 10% on C (10 mg) in methanol (4 mL) was vigorously stirred
under H.sub.2 atmosphere (14 psi). The mixture was stirred for 2
hours at room temperature, filtered through Celite and evaporated
to afford 55 as an oil. Flash chromatography of the residue gave 55
(88 mg, 88%).
[0541] .sup.1H NMR (300 MHz, CDCl.sub.3); .delta. ppm 8.10 (d, 2H,
J=8.0 Hz), 7.35 (m, 7H), 2.73 (m, 4H), 2.00 (m, 2H)
Step 3: N-Hydroxy-4-(3-phenylpropyl)-benzamide (56)
[0542] Following the same procedure as described in Example 21,
Steps 2-3, but substituting compound 55 for compound 43, the title
compound was obtained as a beige solid. (24 mg, 26% yield)
[0543] .sup.1H NMR (300 MHz, CD.sub.3OD); .delta. (ppm) 7.63 (d,
2H, J=8.0 Hz); 7.38-7.05 (m, 7H), 2.63 (m, 4H), 1.91 (m, 2H)
Example 25
N-Hydroxy-4-(4-phenylbutyl)-benzamide (61)
##STR00077##
[0544] Step 1: 4-(1-butenyl-4-phenyl)-benzoic
acid/4-(2-butenyl-4-phenyl)-benzoic acid (57/58)
[0545] Under nitrogen atmosphere in a 25 mL round bottomed flask
were mixed: 4-phenyl-1-butene (568 .mu.L, 3.8 mmol), 4-bromobenzoic
acid (634 mg, 3 2 mmol), tris(dibenzylideneacetone)dipalladium(0)
(87 mg, 0.1 mmol), tri-o-tolylphosphine (58 mg, 0.2 mmol),
triethylamine (1.1 mL, 7.9 mmol) in N,N-dimethylformamide (7 mL,
0.5 M solution). The mixture was stirred for 22 hours at
100.degree. C. Then, the resulting suspension was cooled to room
temperature, filtered through Celite and rinsed with ethyl acetate.
The filtrate was acidified with 1N HCl, the phases were separated
and the aqueous layer was extracted with ethyl acetate. The
combined organic layers were washed with water, brine, dried over
MgSO.sub.4, filtered and concentrated. The resulting solid was
triturated with hexane:dichloromethane (9:1) to give 367 mg (46%)
of beige solid 57/58.
[0546] .sup.1H NMR (300 MHz, (CD.sub.3).sub.2CO): .delta. (ppm)
2.50-2.60 (m, 2H), 2.80 (t, 2H, J=9.0 Hz), 6.40-6.50 (m, 2H),
7.12-7.35 (m, 5H), 7.41 (d, 2H, J=9.0 Hz), 7.92 (d, 2H, J=9.0
Hz).
Step 2: 4-(4-phenylbutyl)-benzoic acid (59)
[0547] Following the procedure described in Example 24, Step 2, but
substituting compound 57/58 for compounds 54, the title compound
was obtained as a white solid in 92% yield.
[0548] .sup.1H NMR (300 MHz, CD.sub.3OD); .delta. (ppm) 1.60-1.75
(m, 4H), 2.65 (t, 2H, J=9.0 Hz), 2.72 (t, 2H, J=9.0 Hz), 7.12-7.30
(m, 5H), 7.33 (d, 2H, J=9.0 Hz), 7.96 (d, 2H, J=9.0 Hz)
Step 3: 4-(4-phenylbutyl)-N--(O-tetrahydropyranyl)-benzamide
(60)
[0549] Under nitrogen atmosphere in a 25 mL round bottomed flask,
to 4-(4-phenylbutyl)benzoic acid 59 (341 mg, 1.3 mmol) in 5 mL of
N,N-dimethylformamide (0.3 M solution) was added the
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (308
mg, 1.6 mmol) and the 1-hydroxybenzotriazole hydrate (272 mg, 2.0
mmol) at room temperature. The mixture was stirred for 30 minutes
then, the 2-(tetrahydropyranyl)hydroxylamine (235 mg, 2.0 mmol) was
added and the mixture was stirred for 4 days. The
N,N-dimethylformamide was removed under vacuum, the resulting oil
was dissolved in ethyl acetate, washed with water and brine, dried
over MgSO.sub.4, filtered and concentrated to give 95% yield of
crude title compound 60.
[0550] .sup.1H NMR (300 MHz, CD.sub.3OD); .delta. (ppm) 1.50-1.75
(m, 10H), 2.65 (t, 2H, J=9.0 Hz), 2.72 (t, 2H, J=9.0 Hz), 3.51 (d,
1H, J=15 Hz), 4.05 (t, 1H, J=15 Hz), 5.05 (s, 1H), 7.10-7.35 (m,
7H), 7.75 (d, 2H, J=9.0 Hz), 10.60 (s, 1H)
Step 4: N-Hydroxy-4-(4-phenylbutyl)-benzamide (61)
[0551] Under nitrogen atmosphere, to the crude oil in a 25 mL round
bottomed flask, were added 5 mL of methyl alcohol (0.3 M solution)
and camphorsulfonic acid (333 mg, 1.4 mmol). The mixture was
stirred for 2 hours at room temperature. The methyl alcohol was
removed under vacuum without heating and the resulting oil was
purified by flash chromatography eluting methyl alcohol and
dichloromethane (1:19). The solid was with hexane:dichloromethane
(9:1) to give 212 mg (59%) of beige solid 61.
[0552] .sup.1H NMR (300 MHz, (CD.sub.3).sub.2CO): .delta. 1.66 (m,
4H), 2.65 (t, 2H, J=7.2 Hz), 2.70 (t, 2H, J=7.1 Hz), 7.15-7.31 (m,
7H), 7.75 (d, 2H, J=7.8 Hz), 8.18 (broad s, 1H), 10.68 (broad s,
1H).
[0553] .sup.13C NMR (75.46 MHz, (CD.sub.3).sub.2CO): .delta. 31.6
(t), 31.8 (t), 36.1 (t), 36.2 (t), 2.times.126.4 (d), 127.8 (d),
2.times.129.1 (d), 2.times.129.2 (d), 2.times.129.3 (d), 130.6 (s),
143.3 (s), 147.3 (s), 165.9 (s).
Example 26
N-Hydroxy-3-(3-phenylpropyl)-benzamide (64)
Step 1: 3-(3-phenylpropenyl)-benzoic acid (62)
[0554] Following the same procedure as described in Example 24,
step 1, but substituting
[0555] 4-bromobenzoic acid for 3-bromobenzoic acid, the title
compound was obtained as mixture of olefins. The mixture was
submitted to the next step without purification.
[0556] .sup.1H NMR (300 MHz, CDCl.sub.3); .delta. (ppm); 3.6 (dd,
2H, CH.sub.2); 6.4 (dd, 2H, vinylic); 7.0-7.5 (m, 8H, CHAr); 8.0
(s, 1H, CHAr)
Step 2: 3-(3-phenylpropyl)-benzoic acid (63)
[0557] Following the same procedure as described in Example 24,
Step 2, but substituting compound 62 for compound 54, the title
compound was obtained in 52% yield and submitted to the next step
without further purification.
[0558] .sup.1H NMR (300 MHz, CDCl.sub.3); .delta. (ppm); 2.0 (m,
2H, CH.sub.2); 2.7 (m, 4H, 2CH.sub.2); 7.0-7.4 (m, 8H, CHAr); 8.0
(s, 1H, CHAr)
Step 3: N-Hydroxy-3-(3-phenylpropyl)-benzamide (64)
[0559] Following the procedure described in Example 25, Step 3-4,
but substituting compound 63 for compound 59, the title compound
was obtained. Purification by flash chromatography using
CH.sub.2Cl.sub.2: MeOH (9.5:0.5) gave compound 64 in 20% yield.
[0560] .sup.1H NMR (300 MHz, DMSO-d.sub.6); .delta. 1.8 (m, 2H,
CH.sub.2); 2.8 (m, 4H, CH.sub.2); 7.0-7.4 (m, 7H, CHAr); 7.6 (s,
CHAr); 9.0 (s, NH); 11.2 (s, OH)
Example 27
N-Hydroxy-3-(2-phenylethyl)-benzamide (68)
##STR00078##
[0561] Step 1: 3-(2-phenylethenyl)-benzoic acid (65/66)
[0562] A 1.0 M solution of lithium bis(trimethylsilyl) amide (3.3
mL, 3.3 mmol) in THF was added to a stirred suspension of
benzyltriphenylphosphonium bromide (1.44 g, 3.6 mmol) in THF (35
mL) at 0.degree. C. The resulting orange solution was added via
cannula to a mixture of 3-carboxybenzaldehyde (500 mg, 3.3 mmol)
and lithium bis(trimethylsilyl)amide (3.3 mL, 3.3 mmol) in THF (10
mL). The mixture was stirred overnight at room temperature. A 1N
solution of HCl (75 mL) and ethyl acetate (75 mL) were added and
the separated aqueous layer was extracted with ethyl acetate
(3.times.50 mL), dried (MgSO.sub.4 anh.) filtered and evaporated.
The residue was purified by HPLC (10:95 CH.sub.3CN: H.sub.2O, TFA
0.1%) to afford 130 mg of the title compound (17%)
[0563] .sup.1H NMR (300 MHz, CDCl.sub.3); .delta. (ppm) (1:1) E:Z
mixture 8.22 (s, 1H), 7.98 (s, 1H), 7.90-7.10 (m, 16H), 6.70 (d,
1H, J=15.0 Hz), 6.62 (d, 1H, J=15.0 Hz)
Step 2: 3-(2-phenylethyl)-benzoic acid (67)
[0564] Following the same procedure as described in Example 24,
Step 2, but substituting compounds 65/66 for compound 54, the title
compound was obtained quantitatively.
[0565] .sup.1H NMR (300 MHz, CDCl.sub.3); .delta. (ppm) 2.98 (m,
4H); 7.30 (m, 7H); 7.99 (m, 2H)
Step 3: N-Hydroxy-3-(2-phenylethyl)-benzamide (68)
[0566] Following the same procedure as described in Example 25,
Step 3 and 4, but substituting compound 67 for compound 59, the
title compound was obtained in 22% yield.
[0567] .sup.1H NMR (300 MHz, DMSO-d.sub.6); .delta. (ppm) 2.82 (s,
4H); 7.03-7.08 (m, 8H); 7.62 (s, 1H); 8.98 (br. s, 1H); 11.15 (br.
s, 1H)
Example 28
N-Hydroxy-4-(2-thiophenyl)-ethyl benzamide (70)
##STR00079##
[0568] Step 1: 4-(2-thiophenyl)-ethyl benzoic acid (69)
[0569] According to the published procedure (Gareau et al., Tet.
Lett., 1994, 1837), under nitrogen atmosphere in a 50 mL round
bottomed flask containing 4-vinylbenzoic acid (1.0 g, 6.75 mmoles)
in 10 mL of benzene (0.7 M) was added benzenethiol (797 .mu.L, 7.76
mmoles) followed by VAZO.TM. (Aldrich Chemical Company, 495 mg,
2.02 mmoles). The mixture was stirred for 12 hours at reflux. The
resulting solution was cooled at room temperature and the solvent
was evaporated under vacuo. The solid was purified by trituration
using hexane and dichloromethane to afford 1.94 g (85%) of white
solid.
[0570] .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. 3.01 (t, 2H,
J=8.4 Hz), 3.28 (dd, 2H, J=7.2, 7.8 Hz), 7.21 (tt, 1H, J=1.2, 7.2
Hz), 7.34 (t, 2H, J=8.1 Hz), 7.38-7.43 (m, 1H), 7.41 (d, 2H, J=8.4
Hz), 7.97 (d, 2H, J=8.1 Hz).
Step 2: N-Hydroxy-4-(2-thiophenyl)-ethyl benzamide (70)
[0571] Under nitrogen atmosphere in a 50 mL round bottomed flask
containing 4-(2-thiophenyl)-ethyl benzoic acid (600 mg, 2.32
mmoles) in 12 mL of N,N-dimethylformamide (0.2 M) was added
1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (579
mg, 3.02 mmoles) and 1-hydroxybenzotriazole hydrate (377 mg, 2.79
mmoles) at room temperature. The mixture was stirred 30 minutes
then, hydroxylamine hydrochloride (242 mg, 3.48 mmoles) and
triethylamine (971 .mu.L, 6.97 mmoles) was added and the mixture
was stirred for 12 hours at 50.degree. C. The N,N-dimethylformamide
was removed under vacuo and the resulting oil was dissolved in
ethyl acetate, washed with water, saturated sodium hydrogen
carbonate solution, water and brine. The organic layer was dried
over anhydrous magnesium sulfate, filtered and concentrated under
vacuo. The crude solid was purified by trituration using hexane and
dichloromethane to afford 450 mg (71%) of a beige solid.
[0572] RP-HPLC (Hewlett-Packard 1100, column C18 HP 4.6.times.250
mm, flow 1 mL/min, 10-95% CH.sub.3CN/H.sub.2O in 42 min with 0.1%
TFA); Purity: 95.8% (220 nm), 93.2% (254 nm).
[0573] .sup.1H NMR (300.072 MHz, (CD.sub.3).sub.2CO): .delta. 2.98
(t, 2H, J=7.2 Hz), 3.26 (dd, 2H, J=6.6, 8.4 Hz), 7.21 (tt, 1H,
J=1.5, 6.9 Hz), 7.31-7.42 (m, 6H), 7.77 (d, 2H, J=9.3 Hz), 8.08
(broad s, 1H), 10.69 (broad s, 1H).
[0574] .sup.13C NMR (75.46 MHz, (CD.sub.3).sub.2CO): .delta. 34.8
(t), 35.9 (t), 126.7 (d), 127.9 (d), 2.times.129.6 (d),
2.times.129.7 (d), 2.times.129.9 (d), 131.3 (s), 137.3 (s), 145.0
(s).
[0575] Elemental Analysis; Calc for
C.sub.15H.sub.15O.sub.2NS.times.0.1H.sub.2O: % C=75.31, % H=7.14, %
N=5.17. Found: % C=75.2.+-.0.1, % H=7.41.+-.0.07, %
N=5.17.+-.0.01.
N-Hydroxy-4-(2-benzenesulfonyl)-ethyl benzamide (73)
Step 1: 4-(2-benzenesulfonyl)-ethyl benzoic acid (72)
[0576] Under nitrogen atmosphere in a 100 mL round bottomed flask
containing 4-(2-thiophenyl)-ethyl benzoic acid (69) (600 mg, 2.32
mmoles) in 20 mL of dichloromethane (0.1 M) at 0.degree. C. was
added portionwise 3-chloroperbenzoic acid (Aldrich Chemical Co.,
57-86% pure solid by, 2 g, 6.97 mmoles), as described by Nicolaou
et al., J. Am. Chem. Soc., 114: 8897 (1992). The mixture was
allowed to reach room temperature and was stirred for 1 hour.
Dimethyl sulfide (5 mL) was added, the mixture was diluted in
dichloromethane and washed 3 times with water. The organic layer
was dried over anhydrous magnesium sulfate, filtered and the
solvent were evaporated in vacuo to afford 3 g of white solid. This
mixture of 3-chlorobenzoic acid and the desired
4-(2-benzenesulfonyl)-ethyl benzoic acid was placed in a 125 mL
Erlenmeyer flask, dissolved in 30 mL of dichloromethane and treated
with an excess of freshly prepared diazomethane solution in diethyl
ether (0.35 M). Nitrogen was bubbled to removed the excess of
diazomethane and solvents were evaporated under vacuum. The
resulting solid was purified by flash chromatography, eluting with
20% ethyl acetate: 80% hexane to afford 341.6 mg (48%) of the
corresponding ester. Saponification of this ester was done using
the same procedure as described in Example 1, step 2, to afford
312.4 mg (96%) of 4-(2-benzenesulfonyl)-ethyl benzoic acid (72)
[0577] .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. 3.06-3.11 (m,
2H), 3.56-3.61 (m, 2H), 7.37 (d, 2H, J=8.4 Hz), 7.67 (tt, 2H,
J=1.5, 7.2 Hz), 7.76 (tt, 1H, J=1.2, 7.5 Hz), 7.93 (d, 2H, J=8.7
Hz), 7.97 (dd, 2H, J=1.8, 6.9 Hz).
Step 2: N-Hydroxy-4-(2-benzenesulfonyl)-ethyl benzamide (73)
[0578] Following the procedure described for
N-hydroxy-4-(2-thiophenyl)-ethyl benzamide, but substituting
4-(2-benzenesulfonyl)-ethyl benzoic acid for 4-(2-thiophenyl)-ethyl
benzoic acid, the title compound was obtained as a beige solid.
[0579] RP-HPLC (Hewlett-Packard 1100, column C18 HP 4.6.times.250
mm, flow 1 mL/min, 10-95% CH.sub.3CN/H.sub.2O in 42 min with 0.1%
TFA); Purity: 98.8% (220 nm), 97.6% (254 nm).
[0580] .sup.1H NMR (300.072 MHz, (CD.sub.3).sub.2CO): .delta. 2.98
(t, 2H, J=7.2 Hz), 3.26 (dd, 2H, J=6.6, 8.4 Hz), 7.21 (tt, 1H,
J=1.5, 6.9 Hz), 7.31-7.42 (m, 6H), 7.77 (d, 2H, J=9.3 Hz), 8.08
(broad s, 1H), 10.69 (broad s, 1H).
[0581] .sup.13C NMR (75.46 MHz, (CD.sub.3).sub.2CO): .delta. 25.2
(t), 34.3 (t), 55.6 (t), 128.0 (d), 2.times.128.8 (d), 129.4 (d),
2.times.130.2 (d), 131.1 (s), 134.5 (d), 140.7 (s), 145.5 (s),
165.8 (s).
N-Hydroxy-4-(2-benzenesulfoxide)-ethyl benzamide (71)
[0582] According to the procedure described by Van Der Borght et
al., J. Org. Chem., 65: 288 (2000), under anitrogen atmosphere in a
10 mL round bottomed flask containing
N-hydroxy-4-(2-thiophenyl)-ethyl benzamide (70) (50 mg, 0.18 mmol)
in 2 mL of methanol (0.1 M) was added tellurium dioxide (3 mg,
0.018 mmol) followed by a solution 35% in water of hydrogen
peroxide (32 .mu.L, 0.36 mmol). The mixture was stirred for five
days and then brine was added. The aqueous layer was extracted 3
times with ethyl acetate and the combined organic layers were dried
over anhydrous magnesium sulfate, filtered and the solvent were
evaporated under vacuo. The resulting solid (43.3 mg) was purified
by trituration using acetonitrile to afford 10 mg (20%) of beige
solid.
[0583] RP-HPLC (Hewlett-Packard 1100, column C18 HP 4.6.times.250
mm, flow 1 mL/min, 10-95% CH.sub.3CN/H.sub.2O in 42 min with 0.1%
TFA); Purity: 98.8% (220 nm), 97.9% (254 nm).
[0584] .sup.1H NMR (300.072 MHz, (CD.sub.3).sub.2CO): .delta.
2.76-2.91 (m, 1H), 3.00-3.29 (m, 3H), 7.34 (d, 2H, J=8.4 Hz),
7.55-7.62 (m, 3H), 7.70 (dd, 2H, J=1.5, 8.1 Hz), 7.76 (d, 2H, J=8.1
Hz), 8.08 (broad s, 1H), 10.70 (broad s, 1H).
[0585] .sup.13C NMR (75.46 MHz, (CD.sub.3).sub.2CO): .delta. 28.3
(t), 57.8 (t), 2.times.124.8 (d), 128.0 (d), 2.times.129.6 (d),
2.times.130.0 (d), 131.5 (d), 144.1 (s), 145.7 (s).
Example 29
N-Hydroxy-3-[4-(3-phenylpropyl)-phenyl]-propanamide (77)
##STR00080##
[0586] Step 1: 3-(4-bromophenyl)-propanoic acid (74)
[0587] Under nitrogen atmosphere in a 250 mL round bottomed flask
containing 4-bromocinnamic acid (5.0 g, 22 mmoles) in 45 mL of
N,N-dimethylformamide (0.5 M) was added benzenesulfonylhydrazide
(7.6 g, 44 mmoles). The mixture was stirred at reflux for 12 hours.
The solution was cooled at room temperature, aqueous saturated
ammonium chloride was added and the aqueous layer was extracted
with ethyl acetate 3 times. Combined organic layers were washed
with water and brine, dried over anhydrous magnesium sulfate,
filtered and concentrated under vacuo. The resulting solid was
purified by flash chromatography eluting with 5% methanol:95%
dichloromethane to afford 3.66 g (73%) of beige solid.
[0588] .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. 2.66 (t, 2H,
J=7.5 Hz), 2.91 (d, 2H, J=7.5 Hz), 7.08 (d, 2H, J=8.4 Hz), 7.41 (d,
2H, J=8.4 Hz).
Step 2: N-Hydroxy-3-(4-bromophenyl)-propanamide (75)
[0589] Following a procedure analogous to that described for the
preparation of 70, 1.54 g (39%) of the title compound was
obtained.
[0590] .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. 2.39 (t, 2H,
J=7.8 Hz), 2.89 (d, 2H, J=7.2 Hz), 7.18 (d, 2H, J=8.1 Hz), 7.42 (d,
2H, J=8.7 Hz), 8.18 (broad s, 1H), 9.98 (broad s, 1H).
Step 3: N-Hydroxy-3-[4-(3-phenyl-1-propenyl)-phenyl]-propanamide
and N-Hydroxy-3-[4-(3-phenyl-2-propenyl)-phenyl]-propanamide
(76)
[0591] Following a procedure analogous to that described in Example
25, step 1, but substituting
N-hydroxy-3-(4-bromophenyl)-propanamide (75) (250 mg, 1.02 mmol)
for 4-bromobenzoic acid and allyl benzene (163 .mu.L, 1.2 mmol) for
4-phenyl-1-butene, to yield 155.4 mg (54%) of the mixed title
compounds.
[0592] .sup.1H NMR (300 MHz, CDCl.sub.3): .delta. 2.39 (m, 2H),
2.88 (t, 2H, J=8.4 Hz), 3.51 (t, 2H, J=8.1 Hz), 6.32-6.53 (m, 2H),
7.14-7.44 (m, 9H), 8.60 (broad s, 1H), 10.04 (broad s, 1H).
Step 4: N-Hydroxy-3-[4-(3-phenylpropyl)-phenyl]-propanamide
(77)
[0593] Following a procedure analogous to that described in Example
24, step 2, but substituting the mixture of
N-hydroxy-3-[4-(3-phenyl-1-propenyl)-phenyl]-propanamide and
N-hydroxy-3-[4-(3-phenyl-2-propenyl)-phenyl]-propanamide (155 mg,
0.55 mmol) for olefins 54, 155.4 mg (99%) of the title compound was
obtained.
[0594] RP-HPLC: (Hewlett-Packard 1100, column C18 HP 4.6.times.250
mm, flow 1 mL/min, 10-95% CH.sub.3CN/H.sub.2O in 42 min with 0.1%
TFA); Purity: 99.9% (220 nm) (2 peaks but same compound proven by
LCMS, 99.9% (254 nm). .sup.1H NMR (300.072 MHz,
(CD.sub.3).sub.2CO): .delta. 1.91 (quintuplet, 2H, J=8.1 Hz), 2.38
(t, 2H, J=7.8 Hz), 2.61 (q, 4H, J=9.6 Hz), 2.87 (t, 2H, J=7.2 Hz),
7.12-7.29 (m, 9H), 8.42 (broad s, 1H), 10.01 (broad s, 1H).
[0595] .sup.13C NMR (75.46 MHz, (CD.sub.3).sub.2CO): .delta. 26.3
(t), 28.7 (t), 29.8 (t), 30.3 (t), 30.7 (t), 121.1 (d),
3.times.123.7 (d), 3.times.123.8 (d), 133.9 (s), 133.4 (s), 137.8
(s), 164.9 (s).
[0596] Elemental Analysis; Calc for
C.sub.18H.sub.21O.sub.2N.times.0.1H.sub.2O: % C=75.81, % H=7.49, %
N=4.91. Found: % C=75.7.+-.0.3, % H=7.54.+-.0.02, %
N=4.85.+-.0.03.
Example 30
##STR00081##
[0597] Step 1: Ethyl 3-(4-nitrophenyl),2-isopropyl propanoate
(78)
[0598] To a precooled solution of diisopropylamine (34 7 mmol) in
THF (30 mL) under nitrogen was added dropwise a 1.0 M solution of
n-butyllithium (33.3 mmol). The resulting light yellow solution was
stirred at -78.degree. C. over 30 minutes and transferred via
canula to a precooled (-78.degree. C.) solution of ethyl
isovalerate (34.7 mmol) in THF (50 mL). The mixture was stirred at
-78.degree. C. over 1 hour and a 4-nitrobenzyl bromide (13.9 mmol)
solution in THF (20 mL) at room temperature was transferred
dropwise via canula to the enolate solution which turned deep red.
The mixture was stirred over 15 minutes and the reaction was
quenched with aqueous saturated ammonium chloride solution
(NH.sub.4Cl). The mixture was allowed to warm to room temperature
over 1 hour and turned brown upon warming. It was poured into a
large volume of saturated NH.sub.4Cl solution and the layers were
separated. The aqueous layer was extracted twice with diethyl ether
and the combined organic layers were washed with brine, dried over
magnesium sulfate and concentrated in vacuo. The residue was
purified by flash chromatography on silica gel using ethyl acetate
and hexanes (10:90) as the eluent, yielding 73% of the pure title
compound 78 as a light yellow oil.
Step 2: Ethyl 3-(4-aminophenyl),2-isopropyl propanoate (79):
[0599] To a hydrogen flushed (vacuum/H.sub.2, 3 times) solution of
1 (1.88 mmol) in methanol (10 mL) was added 10% palladium on
charcoal (0.018 mmol) previously quenched with methanol in a
separate flask. The black heterogeneous resulting mixture was
stirred at room temperature under hydrogen atmosphere (1 atm) over
20 hours. The hydrogen was then evacuated by vacuum and replaced
with air. Then, the mixture was filtered through celite, rinsing
with methanol while making sure the pad never gets dry. The
filtrate was concentrated to a red oil. The residue was purified by
flash chromatography on silica gel using ethyl acetate and hexanes
(30:70) as the eluent, yielding 73% of the pure title compound 79
as a light red oil.
Steps 3-5: (81)
[0600] Compound 79 was coupled with benzenesulfonyl chloride in the
presence of triethylamine according to the procedure described in
Example 1, step 1, to afford the sulfonamide 80. Ester hydrolysis
and coupling with hydroxylamine were then accomplished as described
in Example 28 to afford the hydroxamic acid 81.
[0601] .sup.1H NMR: (Acetone-d.sub.6) .delta. (ppm): 9.76 (bs, 1H),
8.83 (bs, 1H), 7.74 (d, J=8.2 Hz, 2H), 7.59-7.49 (m, 3H), 7.04 (s,
4H), 2.83-2.73 (m, 3H), 1.83 (sext, J=6.9 Hz, 1H), 1.00 (d, J=6.9
Hz, 3H), 0.93 (d, J=6.9 Hz, 3H).
[0602] HRMS: 344.1195 (M.sup.+-H.sub.2O) (calc.);
344.1200.+-.0.0010 (found).
Example 31
##STR00082##
[0604] Compound 82 was obtained in good yield from commercially
available bromoaminopyridine through a palladium catalyzed coupling
with tert-butyl acrylate. Treatment of 82 with
4-phenylbenzenesulfonyl chloride afforded a mixture of sulfonamide
84 and bis-sulfonamide 83, which was converted to 84 upon
chromatographic isolation followed by basic methanolysis. Acidic
cleavage of the t-butyl ester was effected by treatment of 84 with
aqueous formic acid and a tert-butyl cation scavenger to afford the
acrylic acid 85 in quantitative yield. Finally, coupling of 85 with
o-phenylenediamine in the presence of
benzotriazol-1-yloxytris(dimethylamino)phosphonium
hexafluorophosphate (BOP) afforded the anilide 86.
[0605] Data for 86:
[0606] .sup.1H NMR: (300.07 MHz; CD.sub.3OD): .delta. (ppm): 8.23
(d, J=1.9, 1H); 8.03 (bd, J=8.5; 2H); 7.96 (dd, J=1.9, 9.1; 1H);
7.76 (bd, J=8.5, 2H); 7.63 (dd, J=1.4, 8.2); 7.53 (J=15.5; 1H),
7.48-7.36 (m, 3H); 7.29 (d, J=9.1, 1H) 7.18 (dd, J=1.4, 8.0, 1H);
7.03 (dt, J=1.4, 7.8, 1H); 6.86 (d, J=1.4, 7.9, 1H) 6.76 (d,
J=15.6, 1H) 6.75-6.69 (m, 1H); 4.85 (bs, 4H).
[0607] .sup.13C NMR: (75.5 MHz; CD.sub.3OD) .delta. (ppm): 166.4;
154.7; 146.9; 146.2; 143.1; 141.1; 140.6; 138.6; 137.9; 130.1;
129.5; 128.8; 128.5; 128.3; 126.7; 125.6; 125.0; 122.1; 120.8;
119.5; 118.6; 114.9
[0608] MS: calc for C.sub.26H.sub.22O.sub.3N.sub.4S: 470.556.
found: 471.5 for [M+H] (low resolution MS).
[0609] By procedures analogous to those described in Examples 1-31
above, the following compounds were synthesized:
##STR00083##
[0610] .sup.1H NMR: (300 MHz, CD.sub.3OD): .delta.=7.76-7.74 (1H,
m), 7.58-7.48 (4H, m), 7.22 (2H, d, J=7.5 Hz), 7.10 (1H, t, J=5.1
Hz), 6.41 (1H, d broad, J=14.7 Hz).
##STR00084##
[0611] .sup.1H NMR: (300 MHz, CD.sub.3OD): .delta.=7.79 (2H, d,
J=8.1 Hz), 7.56-7.46 (5H, m), 7.17 (2H, d, J=8.1 Hz), 6.39 (1H, d,
J=15.9 Hz).
[0612] Analysis:
C.sub.15H.sub.13N.sub.2O.sub.4SCl.times.0.1H.sub.2O, .times.0.3 TFA
Found: C=48.26%, H=3.58%, N=6.97%, S=7.86%. Calc.: C=48.19%,
H=3.50%, N=7.20%, S=8.25%.
##STR00085##
[0613] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.85 (1H, s
br), 10.70 (1H, s br), 8.99 (1H, s), 8.37 (2H, d, J=9 Hz), 8.01
(2H, d, J=9 Hz), 7.44 (2H, d, J=8.7 Hz), 7.33 (1H, d, J=15.3 Hz),
7.12 (2H, d, J=8.4 Hz), 6.31 (1H, d, J=15.9 Hz).
[0614] Analysis: C.sub.15H.sub.13N.sub.3O.sub.6S.times.0.4H.sub.2O,
.times.0.3 TFA Found: C=46.39%, H=3.49%, N=10.44%, S=7.92%. Calc.:
C=46.29%, H=3.51%, N=10.38%, S=7.92%.
##STR00086##
[0615] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.70 (1H, s
br), 10.33 (1H, s br), 8.99 (1H, s br), 7.44-7.26 (5H, m), 7.12
(2H, d, J=8.7 Hz), 7.06 (1H, d, J=8.4 Hz), 6.30 (1H, d, J=16.2 Hz),
3.78 (3H, s), 3.75 (3H, s)
[0616] Analysis: C.sub.17H.sub.18N.sub.2O.sub.6S.times.0.2H.sub.2O
Found: C=53.56%, H=5.03%, N=7.71%, S=8.01%. Calc.: C=53.45%,
H=4.86%, N=7.33%, S=8.39%.
##STR00087##
[0617] .sup.1H NMR: (CD.sub.3OD) .delta. (ppm): 7.78 (d, J=7.1 Hz,
1H), 7.56-7.45 (m, 3H), 7.24 (d, J=8.5 Hz, 2H), 7.12 (d, J=8.8 Hz,
2H), 7.06 (s, 1H), 2.00 (d, J=1.4 Hz, 3H).
[0618] .sup.13C NMR: (CD.sub.3OD) .delta. (ppm): 135.2, 132.9,
128.1, 127.7, 125.5, 124.6, 124.1, 122.3, 116.8, 115.6, 8.4.
##STR00088##
[0619] .sup.1H NMR: (Acetone-d.sub.6) .delta. (ppm): 9.86 (bs, 1H),
8.86 (bs, 1H), 7.83 (bs, 1H), 7.76 (d, J=6.7 Hz, 1H), 7.62-7.48 (m,
3H), 7.10-7.03 (m, 4H), 2.87-2.79 (m, 3H), 2.56-2.39 (m, 2H), 1.05
(d, J=6.6 Hz, 3H).
[0620] HRMS: 334.0987 (calc.); 334.0991.+-.0.0010 (found).
##STR00089##
[0621] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.94 (1H, s
broad), 10.65 (1H, s broad), 8.95 (1H, s Broad), 8.73-8.71 (1H, m),
8.24-8.21 (2H, m), 8.05 (1H, m), 7.74-7.63 (3H, m), 7.33-7.23 (2H,
m), 7.06-7.04 (2H, m), 6.24 (1H, d, J=15.3).
[0622] Analysis: C.sub.19H.sub.16N.sub.2O.sub.4S.times.0.5H.sub.2O
Found: C=60.31%, H=4.58%, N=7.43%. Calc.: C=60.46%, H=4.54%,
N=7.42%.
##STR00090##
[0623] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.65 (2H, s
broad), 8.48 (1H, s), 8.15-8.08 (2H, m), 8.00 (1H, d, J=7.5 Hz),
7.77 (1H, d, J=9 Hz), 7.70-7.62 (2H, m), 7.39 (2H, d, J=8.4 Hz),
7.28 (1H, d, J=15.6 Hz), 7.15 (2H, d, J=8.4 Hz), 6.26 (1H, d,
J=15.6 Hz).
[0624] Analysis: C.sub.19H.sub.16N.sub.2O.sub.4S.times.0.2H.sub.2O,
.times.0.5 TFA Found: C=56.01%, H=3.94%, N=6.60%, S=7.41%. Calc.:
C=55.99%, H=3.97%, N=6.53%, S=7.47%.
##STR00091##
[0625] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.91 (1H, s),
10.69 (1H, s br), 8.06-7.98 (3H, m), 7.57-7.46 (4H, m), 7.34 (1H,
d, J=15.9 Hz), 7.21 (2H, d, J=8.4 Hz), 6.33 (1H, d, J=15.9 Hz).
##STR00092##
[0626] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=8.69-8.8 (1H,
m), 8.02-8.01 (2H, m), 7.61-7.59 (1H, m), 7.52-7.43 (3H, m), 7.25
(2H, d, J=7.5 Hz), 6.37 (1H, d, J=15.9 Hz).
[0627] Analysis: C.sub.14H.sub.13N.sub.3O.sub.4S.times.0.9 TFA
Found: C=45.36%, H=3.51%, N=9.77%, S=7.09%. Calc.: C=44.97%,
H=3.32%, N=9.96%, S=7.60%.
##STR00093##
[0628] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.91 (1H, s),
10.62 (1H, s br), 8.45 (1H, 8.1 Hz), 8.36 (1H, d, J=8.7 Hz), 8.25
(1H, d, J=6.9 Hz), 7.65-7.59 (2H, m), 7.37-7.34 (2H, m), 7.29-7.23
(2H, m), 7.06 (2H, d, J=8.7 Hz), 6.25 (1H, d, J=15.9 Hz) 2.80 (6H,
s).
##STR00094##
[0629] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.82 (1H, s
br), 9.95 (1H, s br), 9.12 (1H, s br), 7.70 (4H, s), 7.46 (1H, d,
J=15.9 Hz), 6.79 (1H, d, J=8.7 Hz), 6.68 (1H, s), 6.56-6.51 (2H,
m), 3.65 (3H, s), 3.62 (3H, s).
##STR00095##
[0630] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.63 (1H, s),
10.36 (1H, s br), 9.13-9.12 (1H, m), 8.93 (1H, s br), 8.51 (1H, d,
J=8.1 Hz), 8.40 (1H, d, J=7.2 Hz), 8.28 (1H, d, J=8.4 Hz),
7.75-7.70 (2H, m), 7.30-720 (3H, m), 7.09 (2H, d, J=8.4 Hz) 6.21
(1H, d, J=15.9 Hz).
[0631] Analysis: C.sub.18H.sub.15N.sub.3O.sub.4S.times.1.1H.sub.2O
Found: C=55.72%, H=4.45%, N=10.64%, S=6.93%. Calc.: C=55.55%,
H=4.45%, N=10.80%, S=8.24%.
##STR00096##
[0632] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.72 (1H, s
br), 10.07 (1H, s), 7.53-7.51 (2H, m), 7.43-7.34 (4H, m), 7.26-7.19
(4H, m), 6.38 (1H, d, J=15.6 Hz), 4.51 (2H, s).
[0633] Analysis: C.sub.16H.sub.16N.sub.2O.sub.4S.times.0.4 TFA
Found: C=53.60%, H=4.46%, N=7.36%, S=7.81%. Calc.: C=53.38%,
H=4.37%, N=7.41%,0=20.32%, S=8.48%, F=6.03%.
##STR00097##
[0634] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.63 (1H, s
br), 10.56 (1H, s), 8.67 (1H, s), 8.29 (1H, d, J=6.9 Hz), 7.89-7.85
(2H, m), 7.75 (1H, d, J=8.4 Hz), 7.59 (1H, t, J=7.2 Hz), 7.47-7.38
(3H, m), 7.27 (1H, d, J=15.6 Hz), 7.15 (2H, d, J=8.7 Hz), 6.25 (1H,
d, J=15.9 Hz).
##STR00098##
[0635] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.72 (2H, s),
8.98 (1H, s br), 7.97 (4H, s), 7.55 (2H, s), 7.45 (2H, d, J=8.7
Hz), 7.33 (1H, d, J=15.9 Hz), 7.13 (2H, d, J=8.7 Hz), 6.32 (1H, d,
J=15.9 Hz).
##STR00099##
[0636] 1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.75 (2H, m),
7.65-7.64 (1H, m), 7.53-7.45 (4H, m), 7.35 (1H, d, J=16.2 Hz), 7.29
(1H, d, J=3.9 Hz), 7.20 (2H, d, J=8.7 Hz), 7.12 (1H, t, J=3.6 Hz),
6.34 (1H, d, J=15.6 Hz).
[0637] Analysis:
C.sub.17H.sub.14N.sub.2O.sub.4S.sub.3.times.0.1H.sub.2O, .times.1.0
TFA Found: C=43.83%, H=3.26%, N=5.73%, S=18.15%. Calc.: C=43.69%,
H=2.93%, N=5.36%, S=18.42%.
##STR00100##
[0638] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.72 (1H, s),
8.91 (1H, d, J=1.8 Hz), 8.80-8.78 (1H, m), 8.13 (1H, d, J=7.8 Hz),
7.63-7.59 (1H, m), 7.46 (2H, d, J=8.7 Hz), 7.33 (1H, d, J=15.6 Hz),
7.14 (2H, d, J=8.7 Hz), 6.32 (1H, d, J=15.9 Hz).
##STR00101##
[0639] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.54 (1H, s),
7.73 (2H, d, J=8.4 Hz), 7.58 (2H, d, 8.4 Hz), 7.43 (2H, d, J=8.4
Hz), 7.32 (1H, d, J=15.6 Hz), 7.15 (2H, d, J=8.4 Hz), 6.30 (1H, d,
J=15.9 Hz), 1.25 (9H, s).
[0640] Analysis: C.sub.19H.sub.22N.sub.2O.sub.4S.times.0.3H.sub.2O,
0.6 TFA Found: C=54.17%, H=5.25%, N=6.32%, S=6.85%. Calc.:
C=54.12%, H=5.22%, N=6.25%, S=7.15%.
##STR00102##
[0641] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=11.02 (1H, s),
10.70 (1H, s), 8.99 (1H, s br), 8.03 (1H, d, J=1.8 Hz), 7.76-7.67
(2H, m), 7.45 (2H, d, J=8.1 Hz), 7.33 (1H, d, J=15.6 Hz), 7.13 (2H,
d, J=8.4 Hz), 6.31 (1H, d, J=16.2 Hz).
[0642] Analysis:
C.sub.15H.sub.12N.sub.2O.sub.4SCl.sub.2.times.0.3H.sub.2O Found:
C=45.96%, H=3.11%, N=7.21%, S=8.06%. Calc.: C=45.89%, H=3.23%,
N=7.13%, S=8.17%.
##STR00103##
[0643] .sup.1H NMR: (300 MHz, Acetone d.sub.6): .delta.=8.81 (1H,
d, J=8.4 Hz), 8.34 (2H, d, J=7.2 Hz), 8.20 (1H, d, J=8.1 Hz), 8.05
(1H, d, J=7.5 Hz), 7.75-7.59 (4H, m), 7.53-7.41 (4H, m), 7.23-7.07
(4H, m), 6.89-6.86 (2H, m), 6.75 (1H, d, J=15.3 Hz).
[0644] Analysis: C.sub.25H.sub.21N.sub.3O.sub.3S.times.0.4H.sub.2O,
0.6 TFA Found: C=60.68%, H=4.36%, N=8.11%, S=6.15%. Calc.:
C=60.62%, H=4.35%, N=8.09%, S=6.18%.
##STR00104##
[0645] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.7 (1H, s
br), 10.45 (1H, s br), 8.96 (1H, s br), 7.64 (2H, d, J=8.1 Hz),
7.38 (2H, d, J=8.4 Hz), 7.32-7.29 (3H, m), 7.09 (2H, d, J=8.4 Hz),
6.29 (1H, d, J=16.2 Hz), 2.30 (3H, s).
[0646] Analysis: C.sub.16H.sub.16N.sub.2O.sub.4S.times.1.6H.sub.2O,
.times.1.6 TFA Found: C=42.26%, H=3.62%, N=5.45%, S=6.09%. Calc.:
C=42.42%, H=3.86%, N=5.15%, S=5.9%.
##STR00105##
[0647] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.71 (1H, s),
10.67 (1H, s), 9.00 (1H, s br), 7.96 (1H, d, J=2.4 Hz), 7.85 (1H,
d, J=8.4 Hz), 7.69 (1H, dd, J=8.4 Hz and 2.1 Hz), 7.47 (2H, d,
J=8.4 Hz) 7.35 (1H, d, J=15.9 Hz), 7.13 (2H, d, J=8.7 Hz), 6.33
(1H, d, J=15.9 Hz).
[0648] Analysis:
C.sub.15H.sub.12N.sub.2O.sub.4SCl.sub.2.times.0.3H.sub.2O,
.times.0.3 AcOEt Found: C=46.30%, H=3.27%, N=6.56%, S=7.57%. Calc.:
C=46.43%, H=3.61%, N=6.68%, S=7.65%.
##STR00106##
[0649] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=1.65 (1H, s
br), 10.45 (1H, s br), 8.96 (1H, s br), 7.42 (2H, d, J=8.1 Hz),
7.31 (1H, d, J=15.6 Hz), 7.22 (2H, s), 7.01 (2H, d, J=8.1 Hz), 6.30
(1H, d, J=15.9 Hz), 4.24-4.16 (2H, m), 2.93-2.84 (1H, m), 1.18-1.14
(18H, m).
[0650] Analysis: C.sub.24H.sub.32N.sub.2O.sub.4S.times.1.10H.sub.2O
Found: C=62.14%, H=7.17%, N=6.20%, S=6.71%. Calc.: C=62.07%,
H=7.42%, N=6.03%, S=6.9%.
##STR00107##
[0651] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=11.18 (1H, s
br), 10.69 (2H, m), 7.83-7.82 (1H, m), 7.68 (1H, m), 7.43 (2H, d,
J=8.1 Hz), 7.32 (1H, d, J=15.3 Hz), 7.13 (2H, d, J=8.1 Hz), 6.31
(1H, d, J=15.9 Hz).
[0652] Analysis:
C.sub.15H.sub.12N.sub.2O.sub.5SCl.sub.2.times.0.2H.sub.2O,
.times.0.2 TFA Found: C=43.14%, H=3.04%, N=6.54%, S=7.19%. Calc.:
C=43.05%, H=2.96%, N=6.52%, S=7.46%.
##STR00108##
[0653] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.70 (1H, s),
10.65 (1H, s), 9.01 (1H, s br), 7.91 (2H, d, J=8.4 Hz), 7.56 (2H,
d, J=8.4 Hz), 7.45 (2H, d, J=8.1 Hz), 7.33 (1H, d, J=15.6 Hz), 7.13
(2H, d, J=8.1 Hz), 6.31 (1H, J=15.6 Hz).
[0654] Analysis: C.sub.16H.sub.13N.sub.2O.sub.5SF.sub.3.times.0.2
TFA Found: C=46.43%, H=3.33%, N=6.22%, S=7.25%. Calc.: C=46.33%,
H=3.13%, N=6.59%, S=7.54%.
##STR00109##
[0655] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.66 (1H, s
br), 10.37 (1H, s br), 8.56 (1H, s br), 7.69 (2H, d, J=8.7 Hz),
7.39 (2H, d, J=8.1 Hz), 7.30 (1H, d, J=16.2 Hz), 7.10-7.03 (4H, m),
6.27 (1H, d, J=15.9 Hz), 3.77 (3H, s).
[0656] Analysis: C.sub.16H.sub.16N.sub.2O.sub.5S.times.0.7H.sub.2O
Found: C=53.32%, H=5.05%, N=7.98%, S=7.78%. Calc.: C=53.24%,
H=4.86%, N=7.76%, S=8.88%.
##STR00110##
[0657] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.70 (1H, s),
10.66 (1H, s), 8.99 (1H, s), 8.06-7.98 (3H, m), 7.84-7.79 (1H, m),
7.45 (2H, d, J=8.4 Hz), 7.33 (1H, d, J=15.6 Hz), 7.12 (2H, d, J=8.7
Hz), 6.32 (1H, d, J=15.9 Hz).
[0658] Analysis: C.sub.16H.sub.13F.sub.3N.sub.2O.sub.4S Found:
C=49.64%, H=3.30%, N=7.18%. Calc.: C=49.74%, H=3.39%, N=7.25%
##STR00111##
[0659] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.69 (1H, s,
br), 10.47 (1H, s, br), 8.98 (1H, s, br), 7.62 (1H, s), 7.58-7.56
(1H, m), 7.44-7.41 (4H, m), 7.32 (1H, d, J=16.2 Hz), 7.11 (2H, d,
J=8.1 Hz), 6.30 (1H, d, J=15.6 Hz), 2.34 (3H, s).
[0660] Analysis: C.sub.16H.sub.16N.sub.2O.sub.4S.times.0.3 TFA
Found: C=54.64%, H=4.75%, N=7.92%. Calc.: C=54.66%, H=4.59%,
N=7.82%
##STR00112##
[0661] .sup.1H NMR: (300 MHz, MeOD d.sub.4): 7.62-6.61 (m, 13H);
3.81 (broad s, 3H, OCH.sub.3), 3.80 (broad s, 3H, OCH.sub.3),3.26
(broad s, 4H, NH).
[0662] .sup.13C NMR: (75 MHz, MeOD d.sub.4): 167.0 (C.dbd.O);
154.4; 150.5; 143.1; 141.9; 141.0; 132.5; 132.3; 129.9; 128.2;
126.7; 125.2; 122.4; 121.8; 120.8; 119.6; 118.7; 111.9; 110.9; 56.6
(2C, OCH.sub.3).
[0663] Combustion analysis: Calc: 60.91% C, 5.11% H, 9.27% N, 7.07%
S.
[0664] Found: 60.40% C, 5.21% H, 9.16% N, 6.47% S.
[0665] HRMS: Calc: 453.1358. Found: 453.1351.
##STR00113##
[0666] .sup.1H NMR: (Acetone-d.sub.6): .delta. (ppm): 9.25 (bs,
1H), 8.77 (bs, 1H), 7.79 (d, J=8.5 Hz, 2H), 7.61-7.51 (m, 5H),
7.36-7.28 (m, 3H), 6.99-6.93 (m, 1H), 6.86-6.82 (m, 2H), 6.68-6.62
(m, 1H), 4.63 (bs, 2H).
[0667] HRMS: 449.1773 (calc.): 449.1767.+-.0.0013 (found).
##STR00114##
[0668] .sup.1H NMR: (300 MHz, MeOD d.sub.4): 8.00-6.56 (m, 13H);
3.77 (broad s, 3H, OCH.sub.3), 3.74 (broad s, 3H, OCH.sub.3), 3.33
(broad s, 2H, NH), 3.00 (broad s, 1H, NH), 2.88 (broad s, 1H,
NH).
[0669] .sup.13C NMR: (75 MHz, MeOD d.sub.4): 166.2 (C.dbd.O);
150.7; 148.5; 143.2; 141.7; 140.6; 140.5; 131.9; 129.2; 128.9;
128.4; 126.7; 124.9; 119.5; 118.6; 116.4; 113.2; 108.9; 56.6
(OCH.sub.3); 56.4 (OCH.sub.3).
[0670] MS: Calc: 453.1358. Found: 453.1351.
##STR00115##
[0671] .sup.1H NMR: (CD.sub.3OD) .delta. (ppm): 7.68 (d, J=8.2 Hz,
2H), 7.55 (d, J=15.9 Hz, 1H), 7.47 (d, J=8.5 Hz, 2H), 7.30 (d,
J=8.0 Hz, 2H), 7.19-7.12 (m, 3H), 7.03 (t, J=7.1 Hz, 1H), 6.86 (d,
J=8.0 Hz, 1H), 6.75-6.69 (m, 2H), 2.37 (s, 3H).
[0672] HRMS: 407.1304 (calc.): 407.1293.+-.0.0012 (found).
##STR00116##
[0673] .sup.1H NMR: (300 MHz, DMSO-d.sub.6) .delta. 10.6 (s, OH); 9
(s, NH); 7.1-7.8 (m, 14H, CH Ar); 6.2 (d, 1H, J=15 Hz)
##STR00117##
[0674] .sup.1H NMR: (300 MHz, MeODd.sub.4): 7.31-6.62 (m, 11H);
3.72 (broad s, 3H); 3.70 (broad s, 3H); 2.91 (t, 2H; J=7.1 Hz);
2.65 (broad t, 2H, J=7.4 Hz)
[0675] .sup.13C NMR: (75 MHz, MeODd.sub.4): 173.9; 154.0; 150.3;
143.4; 138.6; 137.4; 132.6; 130.2; 128.4; 127.4; 124.6; 123.1;
122.3; 119.3; 118.1; 111.7; 110.9; 56.5 (2C); 38.8; 32.2.
[0676] HRMS: calc: 455.1515. Found: 455.1521.
##STR00118##
[0677] .sup.1H NMR: (300 MHz, DMSO d.sub.6): 7.77 (d, 2H, J=8.8
Hz); 7.51 (d, 2H, J=8.5 Hz); 7.34 (d, 2H, J=8.8 Hz); 7.18 (d, 2H,
J=8.5 Hz); 7.11 (d, 2H, 8.8 Hz); 6.94 (t, 1H, J=7.4 Hz); 6.77
(broad d, 2H, J=7.9 Hz); 6.6 (t, 1H, J=7.4 Hz), 4.95 (broad s, 1H),
3.83 (s, 3H).
[0678] .sup.13C NMR: (75 MHz, DMSO d.sub.6): 162.5; 141.5; 139.2;
138.8; 130.9; 130.2; 128.9; 128.6; 125.7; 124.7; 119.4; 116.2;
115.9; 114.5; 55.6.
[0679] HRMS: Calc: 423.1253. Found: 423.1235.
##STR00119##
[0680] .sup.1H NMR: (300 MHz, DMSO-d.sub.6) .delta. 7.1-7.8 (m,
14H, CH Ar); 6.8-6.9 (m, 4H, CH Ar); 6.3 (d, 1H, J=15 Hz)
Example 32
##STR00120##
[0682] Sulfonamide 124 was prepared by condensation of
4-iodoaniline with benzenesulfonyl chloride. Compound 125 was
quantitatively furnished by a Pd--Cu catalyzed coupling reaction of
124 with propargyl alcohol in basic solvent. Primary alcohol 125
was oxidized to the corresponding carboxylic acid 127 in two steps,
including Dess-Martin periodinane oxidation to afford aldehyde 126,
followed by treatment with sodium chlorite in buffered aqueous
media in the presence of a chlorine scavenger. Acid 127 was
derivatized to the hydroxamic acid 128 by treatment with
hydroxylamine hydrochloride and the coupling reagent EDC in the
presence of N-hydroxybenzotriazole in basic, aprotic media.
Compound 129 was prepared by coupling acid 130 with
o-phenylenediamine as described in Example 31 for compound 86.
[0683] Data for 128:
[0684] .sup.1H NMR: (300.07 MHz; acetone-d.sub.6) .delta. (ppm):
9.4 (bs, 2H); 7.93 (dd, J=1.9, 6.6; 2H); 7.82 (dd, J=1.9, 6.6; 2H);
7.68 (dd, J=1.4, 8.2; 2H); 7.48-741 (m, 5H); 7.35-7.32 (m, 2H);
2.90 (bs, 1H)
[0685] .sup.13C NMR: (75.5 MHz; acetone-d.sub.6) .delta. (ppm):
153.5; 147.2; 141.3; 140.3; 139.5; 134.6; 130.1; 129.5; 128.8;
128.6; 128.3; 120.8; 116.5; 87.7; 81.0.
[0686] MS: calc for C.sub.21H.sub.16O.sub.4N.sub.2S: 392.438.
found: 393.4 for [M+H] (low resolution MS).
[0687] Data for 129:
[0688] .sup.1H NMR: (300.07 MHz; acetone-d.sub.6) .delta. (ppm):
9.43 (bs, 1H); 8.02 (d, J=8.5 Hz; 2H); 7.93 (d, J=8.5 Hz; 2H); 7.90
(d, J=8.5 Hz; 2H); 7.65 (d, J=8.5 Hz; 2H); 7.47-7.34 (m, 7H);
7.21-7.17 (m, 2H); 2.80 (bs, 3H)
[0689] .sup.13C NMR: (75.5 MHz; acetone-d.sub.6) .delta. (ppm):
167.2; 158.6; 146.3; 141.3; 140.9; 139.8; 139.5; 134.2; 131.0;
129.9; 129.8; 129.3; 128.7; 128.6; 128.4; 128.0; 126.8; 125.1;
122.7; 122.6; 120.1
[0690] MS: calc for C.sub.27H.sub.21O.sub.3N.sub.3S: 467.552.
found: 468.5 for [M+H] (low resolution MS).
Example 33
##STR00121##
[0692] Benzylic alcohol 130 was prepared in 53% yield by addition
of 2-lithiofuran to styrene oxide. After protection of the
resulting hydroxyl group with tert-butyldimethylsilyl chloride, the
lithiated species of compound 131 was treated with DMF to afford
the formyl derivative 132. Wadsworth-Horner-Emmons olefination was
effected by treatment of 132 with the sodium enolate of
trimethylphosphonoacetate to afford the key intermediate 133 in 90%
overall yield for the last three steps. Saponification of the
methyl ester with LiOH yielded the acid 134, which in turn was
converted into its hydroxamic acid form 135 by conventional
activation with HOBt/EDC, followed by reaction with hydroxylamine
Fluoride-promoted cleavage of silylated ether gave alcohol 136 in
67% yield.
[0693] Data for 136:
[0694] .sup.1H NMR: (300.07 MHz; acetone-d6) .delta. (ppm): 9.35
(bs, 1H); 7.40-7.15 (m; 6H); 6.56 (d, J=2.9 Hz, 1H); 6.24 (d,
J=15.3 Hz, 1H); 4.96 (t, J=6.2 Hz, 1H); 3.00 (d, J=6.2 Hz, 2H)
[0695] .sup.13C NMR: (75.5 MHz; CD.sub.3OD) .delta. (ppm): 166.6;
156.6; 151.3; 145.2; 129.3; 128.5; 126.9; 116.2; 114.5; 111.0;
73.6; 39.1
Example 34
##STR00122##
[0697] Unsaturated ketoacid 138 was obtained from ester 133 in 73%
overall yield after three consecutive steps, including
saponification (LiOH/H.sub.2O/MeOH/THF), desilylation (TBAF/THF),
and oxidation of benzylic alcohol 137 using Dess-Martin
periodinane. Anilide 139 was obtained by BOP-mediated condensation
of compound 138 with o-phenylenediamine in 83% yield.
[0698] Regioselective hydrogenation of the acrylate moiety in 133
was accomplished by treatment with NaBH.sub.4 in the presence of
NiCl.sub.2, to afford the propionate 140 in high yield. Ketoacid
142 was then obtained in 31% overall yield from 140 by an identical
procedure to that followed in the synthesis of 138 from 133. With
compound 142 in hand, anilide 144 was obtained as described above
(BOP/o-phenylendiamine). The low yield was due to a difficult
purification process. To avoid oxime formation, hydroxamic acid 143
was synthesized from 142 in 73% overall yield over two steps,
including BOP-mediated coupling with
N,O-bistrimethylsilylhydroxylamine, followed by cleavage of
silylated groups under acidic conditions (citric acid/MeOH).
[0699] Data for 139:
[0700] .sup.1H NMR: (300.07 MHz; CDCl.sub.3) .delta. (ppm):
8.02-7.42 (series of multiplets, 7H); 7.34 (bs, 1H); 7.06 (m, 1H);
6.80 (d, J=7.8; 1H); 6.79 (d, J=8.1; 1H); 6.54 (d, J=3.0 Hz, 1H);
6.38 (m, 1H); 6.34 (d, J=3.0 Hz, 1H); 4.37 (s, 2H); 3.90 (bs,
2H)
[0701] .sup.13C NMR: (75.5 MHz; CDCl.sub.3) .delta. (ppm): 194.5;
164.4; 150.9; 150.8; 150.5; 140.5; 135.9; 133.7; 128.7; 128.5;
126.9; 125.0; 124.4; 119.4; 118.0; 117.5; 115.7; 111.3; 38.5
[0702] Data for 143:
[0703] .sup.1H NMR: (300.07 MHz; CDCl.sub.3) .delta. (ppm): 8.99
(bs, 1H); 8.09-7.42 (series of multiplets, 5H); 6.09 (d, J=3.0 Hz,
1H); 6.00 (d, J=3.0 Hz, 1H); 4.35 (s, 2H); 2.95 (t, J=6.60 Hz, 2H);
2.50 (t, J=3.0 Hz, 1H).
[0704] .sup.13C NMR: (75.5 MHz; CDCl.sub.3) .delta. (ppm): 196.2;
162.8; 153.2; 146.8; 134.9; 133.7; 128.7; 128.5; 109.3; 107.1;
38.2; 31.7; 24.2
[0705] Data for 144:
[0706] .sup.1H NMR: (300.07 MHz; CDCl.sub.3) .delta. (ppm):
7.99-7.42 (series of multiplets, 5H); 7.36 (bs, 1H); 7.02 (d,
J=7.8, 2H); 6.73 (d, J=7.8 Hz, 2H); 6.13 (d, J=3.0 Hz, 1H); 6.04
(d, J=3.0 Hz, 1H); 4.30 (s, 2H); 3.70 (bs, 2H); 3.03 (t, J=6.9 Hz,
2H); 2.69 (t, J=6.9 Hz, 2H).
[0707] .sup.13C NMR: (75.5 MHz; CDCl.sub.3) .delta. (ppm): 195.4;
170.7; 153.6; 147.1; 140.9; 136.1; 133.5; 128.7; 128.5; 127.1;
125.7; 124.0; 119.2; 117.8; 109.1; 107.2; 38.4; 35.7; 24.7
Example 35
General Procedure for Synthesis of Urea Compounds
##STR00123##
[0709] To a solution of isocyanate (1.5 mmol) in 15 mL of anhydrous
dichloromethane, was added a solution of 4-anilinylmethylacrylate
(1.5 mmol) in dichloromethane (10 mL). The mixture was stirred at
room temperature for 15 hours. After addition of ammonium chloride
solution the new mixture was extracted from dichloromethane. The
organic layers were combined and washed with ammonium chloride
solution, water, brine and dried over magnesium sulfate. The crude
was then flashed over silica gel using CH.sub.2Cl.sub.2: MeOH
(9.5:0.5) as eluent.
[0710] The following compounds were synthesized according to the
general procedure:
##STR00124##
[0711] .sup.1H NMR: (300 MHz, DMSO-d.sub.6) .delta. 7.5-7.7 (m, 4H,
CH Ar); 7.5 (d, 2H, J=6.6 Hz); 7.3 (d, 2H, J=6.6 Hz); 6.3 (d, 1H,
J=15 Hz)
##STR00125##
[0712] .sup.1H NMR: (300 MHz, DMSO-d.sub.6) .delta. 7.5-8.2 (m, 7H,
CH Ar); 7.5 (d, 2H, J=6.6 Hz); 7.3 (d, 2H, J=6.6 Hz); 6.3 (d, 1H,
J=15 Hz)
##STR00126##
[0713] .sup.1H NMR: (300 MHz, DMSO-d.sub.6) .delta. 7.5-7.7 (m, 3H,
CH Ar); 7.5 (d, 2H, J=6.6 Hz); 7.3 (d, 2H, J=6.6 Hz); 6.3 (d, 1H,
J=15 Hz)
Example 36
[0714] The following additional compounds were prepared by
procedures analogous to those described in the foregoing
Examples:
##STR00127##
[0715] .sup.1H NMR (300.072 MHz, (CD.sub.3).sub.2CO): .delta. 1.99
(m, 2H), 2.79 (t, 2H, J=7.2 Hz), 3.21 (dd, 2H, J=6.8, 7.8 Hz), 7.27
(d, 2H, J=8.1 Hz), 7.65 (t, 2H, J=7.8 Hz), 7.72-7.77 (m, 3H), 7.90
(d, 2H, J=7.2 Hz), 10.77 (broad s, 1H).
[0716] .sup.13C NMR (75.46 MHz, (CD.sub.3).sub.2CO): .delta. 25.2
(t), 34.3 (t), 55.6 (t), 128.0 (d), 2.times.128.8 (d), 129.4 (d),
2.times.130.2 (d), 131.1 (s), 134.5 (d), 140.7 (s), 145.5 (s),
165.8 (s).
##STR00128##
[0717] .sup.1H NMR (300.072 MHz, (CD.sub.3).sub.2CO): .delta.
1.66-1.88 (m, 4H), 2.71 (t, 2H, J=6.3 Hz), 4.34 (d, 1H, J=303 Hz),
4.87 (m, 1H), 7.27 (d, 2H, J=7.8 Hz), 7.44-7.48 (m, 2H), 7.52 (dd,
1H, J=1.5, 9.4 Hz), 7.73 (d, 2H, J=7.8 Hz), 7.83 (s, 1H), 7.83-7.88
(m, 3H), 8.16 (broad s, 1H), 10.67 (broad s, 1H).
[0718] .sup.13C NMR (75.46 MHz, (CD.sub.3).sub.2CO): .delta. 28.3
(t), 36.2 (t), 39.8 (t), 74.0 (d), 125.0 (d), 125.3 (d), 126.2 (d),
126.7 (d), 2.times.127.8 (d), 128.4 (d), 128.5 (d), 128.6 (d),
2.times.129.3 (d), 130.6 (s), 133.7 (s), 134.3 (s), 144.7 (s),
147.4 (s), 165.9 (s).
##STR00129##
[0719] .sup.1H NMR: (300 MHz, DMSO-d6) .delta. 11.2 (s, OH); 9 (s,
NH); 7.6-7.8 (m, 4H, CH Ar); 7-7.4 (m, 5H, CH Ar); 2.8 (m, 4H,
CH.sub.2).
##STR00130##
[0720] .sup.1H NMR: (300 MHz, DMSO-d.sub.6) .delta. 11.2 (s, 1H);
9.0 (s, 1H); 7.7 (m, 6H); 7.34 (m, 5H).
##STR00131##
[0721] .sup.1H NMR: (300 MHz, DMSO-d.sub.6) .delta. 11.2 (s, OH); 9
(s, NH); 7.6-7.8 (m, 4H, CH Ar); 7-6.8 (m, 4H, CH Ar); 2.9 (s, 6H,
2CH.sub.3); 2.8 (m, 4H, CH.sub.2).
##STR00132##
[0722] .sup.1H NMR (300.072 MHz, (CD.sub.3).sub.2CO): .delta. 1.38
(quintuplet, 2H, J=7.5 Hz), 1.60-1.72 (m, 4H), 2.60 (t, 2H, J=7.8
Hz), 2.67 (t, 2H, J=7.5 Hz), 7.15-7.31 (m, 7H), 7.75 (d, 2H, J=8.1
Hz), 8.11 (broad s, 1H), 10.68 (broad s, 1H).
[0723] .sup.13C NMR (75.46 MHz, (CD.sub.3).sub.2CO): .delta. 31.8
(t), 32.1 (t), 36.2 (t), 36.4 (t), 126.4 (d), 127.8 (d),
2.times.129.0 (d), 2.times.129.2 (d), 2.times.129.3 (d), 143.3
(s).
##STR00133##
[0724] .sup.1H NMR (300.072 MHz, (CD.sub.3).sub.2CO): .delta. 1.63
(m, 4H, J=4.5 Hz), 2.37 (t, 2H, J=7.8 Hz), 2.57-2.66 (m, 4H), 2.86
(t, 2H, J=7.5 Hz), 7.10-7.28 (m, 9H), 8.01 (broad s, 1H), 9.98
(broad s, 1H).
[0725] .sup.13C NMR (75.46 MHz, (CD.sub.3).sub.2CO): .delta. 31.0
(t), 2.times.31.9 (t), 35.1 (t), 35.8 (t), 36.2 (t), 126.4 (d),
2.times.129.0 (d), 2.times.129.1 (d), 2.times.129.1 (d), 129.2 (d),
138.8 (s), 141.2 (s), 143.4 (s), 164.1 (s).
##STR00134##
[0726] .sup.1H NMR (300.072 MHz, (CD.sub.3).sub.2CO): .delta.
1.83-1.98 (m, 4H), 2.08-2.14 (m, 2H), 2.56-2.67 (m, 6H), 7.12-7.30
(m, 9H), 9.98 (broad s, 1H).
##STR00135##
[0727] .sup.1H NMR (300.072 MHz, (CD.sub.3).sub.2CO): .delta.
1.60-1.68 (m, 4H), 1.87 (quintuplet, 2H, J=7.5 Hz), 2.03-2.14 (m,
2H), 2.55-2.67 (m, 6H), 7.09-7.28 (m, 9H).
##STR00136##
[0728] .sup.1H NMR (300.072 MHz, (CD.sub.3).sub.2CO): .delta. 2.37
(t, 2H, J=7.2 Hz), 2.78-2.89 (m, 6H), 7.13-7.29 (m, 9H), 7.84
(broad s, 1H), 9.90 (broad s, 1H).
[0729] .sup.13C NMR (75.46 MHz, (CD.sub.3).sub.2CO): .delta. 31.6
(t), 35.1 (t), 38.2 (t), 38.6 (t), 2.times.126.6 (d), 2.times.129.1
(d), 2.times.129.2 (d), 2.times.129.3 (d), 139.4 (s), 140.4 (s),
142.8 (s), 170.1 (s).
##STR00137##
[0730] .sup.1H NMR (300.072 MHz, (CD.sub.3).sub.2CO): .delta. 1.96
(quintuplet, 2H, J=6.0 Hz), 2.69 (t, 2H, J=8.0 Hz), 3.19 (dd, 2H,
J=6.0, 9.0 Hz), 3.38 (s, 2H), 7.09 (d, 2H, J=7.5 Hz), 7.21 (d, 2H,
J=7.5 Hz), 7.66 (t, 2H, J=8.1 Hz), 7.747 (t, 1H, J=6.9 Hz), 7.90
(d, 2H, J=6.6 Hz), 10.08 (broad s, 1H).
[0731] .sup.13C NMR (75.46 MHz, (CD.sub.3).sub.2CO): .delta. 25.5
(t), 34.1 (t), 39.9 (t), 55.7 (t), 2.times.128.8 (d), 130.0 (d),
2.times.130.2 (d), 134.4 (s), 139.9 (s), 140.7 (s), 168.5 (s).
##STR00138##
[0732] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta. 7.77 (broad s,
4H); 7.57 (d, 1H, J=15.7 Hz); 7.35 (d, 1H, J=6.9 Hz); 7.03-6.94 (m,
6H); 6.76 (d, 1H, J=7.1 Hz); 6.59 (d, 1H, J=6.9 Hz); 4.98 (broad s,
2H); 2.19 (s, 3H).
[0733] .sup.13C NMR: (75 MHz, DMSO d.sub.6): .delta. 162.9; 141.6;
139.8; 139.0; 137.6; 134.8; 133.6; 129.6; 128.1; 127.3; 125.9;
125.4; 124.7; 123.2; 120.7; 116.2; 115.9; 20.3.
##STR00139##
[0734] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta. 7.91-7.81 (m,
4H); 7.63-7.58 (m, 5H); 7.48-7.43 (m, 2H); 7.39-7.33 (m, 2H); 7.24
(d, 2H, J=8.5 Hz); 6.97 (dd, 2H, J=9.9, 7.1 Hz); 6.79 (d, 1H, J=7.7
Hz) 6.61 (dd, 1H, J=7.7, 7.1 Hz); 5.01 (broad s, 2H).
[0735] .sup.13C NMR: (75 MHz, DMSO d.sub.6): .delta. 162.9; 141.9;
141.6; 139.8; 139.2; 137.6; 136.9; 135.8; 128.9; 128.3; 127.4;
127.3; 127.2; 126.3; 126.0; 125.5; 124.8; 123.2; 120.4; 116.2;
115.9.
##STR00140##
[0736] .sup.1H NMR: (300 MHz, MeODd.sub.4): .delta. 7.74-7.54 (m,
5H); 7.07-6.96 (m, 4H); 6.55 (d, 1H, J=15.7); 2.25 (s, 3H).
[0737] .sup.13C NMR: (75 MHz, MeODd.sub.4): .delta. 163.5; 141.6;
140.4; 139.5; 136.1; 135.9; 130.6; 129.0; 128.8; 123.1; 121.7;
20.8
##STR00141##
[0738] .sup.1H NMR: (300 MHz, MeODd.sub.4): .delta. 7.83-7.19 (m,
14H); 6.56 (d, 1H, J=15.7 Hz).
[0739] .sup.13C NMR: (75 MHz, MeODd.sub.4): .delta. 165.4; 141.6;
141.4; 140.5; 139.5; 139.0; 137.9; 129.8; 129.2; 128.7; 128.6;
128.2; 127.6; 122.7; 121.7.
Example 37
##STR00142##
[0741] To a solution of carboxylic acid 163 (131 mg, 0.36 mmol),
prepared according to procedures described above, in 6 mL of dry
DMF was added Et.sub.3N (190 .mu.l, 1.37 mmol), followed by the
addition of solid BOP (259 mg; 0.59 mmol). The reaction mixture was
stirred for 10 min. at room temperature and then solid
5-amino-1,3,4-thiadiazole-2-thiol (58 mg, 0.43 mmol) was added.
After being stirred for 12 h, the mixture was diluted with methanol
and concentrated under vacuum. Upon dilution with
CH.sub.2Cl.sub.2/MeOH, crystallization of 164 (150 mg, 87%) from
the crude oil took place.
[0742] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta. 7.85 (broad s,
5H); 7.04-6.58 (m, 4H); 3.69 (s, 3H); 3.67 (s, 3H); 3.38 (broad s,
3H).
[0743] .sup.13C NMR: (75 MHz, DMSO d.sub.6): 163.3; 161.7; 158.7;
148.7; 146.2; 142.0; 140.7; 137.9; 130.1; 128.7; 127.5; 121.4;
113.7; 112.0; 106.6; 55.5; 55.4.
[0744] Following this general procedure, the following thiadiazole
derivatives were prepared from the corresponding carboxylic
acids:
##STR00143##
[0745] .sup.1H NMR: (300 MHz, DMSO-d.sub.6); .delta. (ppm):
7.89-7.72 (series of multiplets, 7H); 7.50-7.05 (series of
multiplets, 6H); 3.32 (broad singlet, 3H)
[0746] .sup.13C NMR: (75 MHz, DMSO-d6); d (ppm): 162.6; 162.3;
144.5; 138.3; 138.3; 138.2; 132.5; 130.1; 129.7; 129.1; 128.6;
127.6; 127.3; 127.1; 120.9; 118.7; 116.8.
[0747] MS: calc. for M+H, 493.6. obs. for M+H, 496.3
##STR00144##
[0748] .sup.1H NMR: (300 MHz, DMSO d.sub.6): 7.87-7.72 (m, 5H),
7.57-7.53 (m, 4H), 7.39 (dd, 2H, J=6.9, 7.7 Hz), 7.30 (d, 1H, J=7.1
Hz), 7.17 (d, 2H, J=8.5 Hz), 6.85 (d, 1H, J=15.9 Hz).
[0749] MS: cal: 495.61. found: 496.6.
[0750] Following an analogous procedure, but substituting
2-amino-5-trifluoro-methyl-1,3,4-thiadiazole for
5-amino-1,3,4,-thiadiazol-2-thiol, the following compound was
prepared:
##STR00145##
[0751] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta. 7.96-7.81 (m,
5H); 7.71-7.48 (m, 4H); 7.38 (dd, 2H, J=7.1, 7.41 Hz); 7.28 (d, 1H,
J=7.1 Hz); 7.19 (d, 2H, J=8.5 Hz); 6.98 (d, 1H, J=15.7 Hz).
[0752] .sup.13C NMR: (75 MHz, DMSO d.sub.6): 192.3; 163.6; 161.6;
142.4; 140.9; 139.2; 138.0; 136.8; 135.9; 129.0; 128.8; 127.4;
127.2; 126.2; 121.2; 120.4.
[0753] MS: cal: 530.55 found: 531.5.
Example 38
##STR00146##
[0755] Coupling of 24 (from Example 15) with o-phenylenediamine in
the presence of benzotriazol-1-yloxytris(dimethylamino)phosphonium
hexafluorophosphate (BOP) afforded the anilide 168.
[0756] By an analogous procedure, the corresponding
para-substituted compound is prepared from 32 (from Example
16).
Example 39
##STR00147##
[0757] Step 1: N-Methyl-4-iodophenylbenzenesulfonamide (169)
[0758] To compound 28 (from Example 18) (500 mg, 1.39 mmol) in DMF
(10 mL) were added at room temperature K.sub.2CO.sub.3 (962 mg,
6.96 mmol), followed by methyl iodide (395 mg, 2.78 mmol). The
resulting reaction mixture was stirred at room temperature for 16
hours. The solvent is then removed and water was added. The
resulting mixture was extracted with ethyl acetate, and the
combined organic phases were dried and concentrated. Purification
by flash chromatography using hexane:ethyl acetate (8:2) afforded
510 mg (98%) of the title compound as a white solid.
[0759] Compound 169 was converted to the hydroxamic acid 170
according to the procedures described in Example 18 for the
preparation of compound 36.
[0760] Data for 170:
[0761] .sup.1H NMR: (300 MHz, DMSO d.sub.6): .delta.=10.76 (1H, s),
9.04 (1H, s), 7.73-7.68 (1H, m), 7.61-7.51 (6H, m), 7.43 (1H, d,
J=15.9 Hz), 7.15 (2H, d, J=8.7 Hz), 6.43 (1H, d, J=16.2 Hz), 3.15
(3H, s).
[0762] Analysis: C.sub.16H.sub.16N.sub.2O.sub.4S.times.0.5H.sub.2O
Found: C=56.36%, H=5.09%, N=8.69%, S=8.33%. Calc.: C=56.29%,
H=5.02%, N=8.21%, S=9.39%.
Example 40
N-hydroxy-2-(4-(4-phenylbutyl)phenyl)acetamide (174)
Step 1: Methyl 2-(4-iodophenyl)acetate
##STR00148##
[0764] A 4M solution of HCl in dioxane (50 mL) was added to
2-(4-iodophenyl)acetic acid (10 g, 38.2 mmol) in MeOH (100 mL) and
the reaction stirred overnight. The solvent was evaporated under
reduced pressure. The residue was purified by silica gel column
chromatography eluting with 0-30% EtOAc in hexanes to afford methyl
2-(4-iodophenyl)acetate in 10.42 g (99%). LRMS: 276.0 (calc) 277.1
(found) (MH).sup.+.
Step 2: Methyl 2-(4-(4-phenylbut-1-ynyl)phenyl)acetate
##STR00149##
[0766] CuI (0.06 equiv, 207 mg, 1.1 mmol) was added to a solution
of Pd(Ph.sub.3P).sub.4 (0.03 equiv, 628 mg, 0.54 mmol) and aromatic
iodide (1 equiv, 5 g, 18 mmol) in Et.sub.2NH:DME (30 mL:30 mL) and
the reaction stirred for 20 min. 4-phenyl-1-butyne (3 equiv, 7.1 g,
54 mmol) was then added dropwise and the reaction stirred for 3 h.
The reaction was concentrated under reduced pressure and then the
residue partitioned between EtOAc (50 mL) and H.sub.2O (50 mL). The
organic phase was separated, dried over Na.sub.2SO.sub.4 filtered
and concentrated. The compound was purified by silica gel flash
column chromatography: 20%-100% EtOAc in hexanes to afford methyl
2-(4-(4-phenylbut-1-ynyl)phenyl)acetate in 4.95 g (98%). LRMS:
278.3 (calc) 279.2 (found) (MH).sup.+.
Step 3: Methyl 2-(4-(4-phenylbutyl)phenyl)acetate
##STR00150##
[0768] 10% Pd/C (20% w/w, 1 g) was added to a solution of the
acetylene (4.9 g, 18 mmol) in MeOH (30 mL). The reaction was then
purged with H.sub.2 and stirred overnight. The solvent was
evaporated and the residue was purified by a silica plug eluting
with 30% EtOAc in hexanes to afford methyl
2-(4-(4-phenylbutyl)phenyl)acetate in 4.99 g (99%). LRMS: 282.3
(calc) 283.1 (found) (MH).sup.+.
Step 4: N-hydroxy-2-(4-(4-phenylbutyl)phenyl)acetamide (174)
##STR00151##
[0770] NaOH (6 equiv, 4.2 g, 106 mmol) was added to a solution of
the methyl ester (1 equiv, 4.9 g, 18 mmol) and aq. NH.sub.2OH (50
equiv, 58 g, 884 mmol) in MeOH (100 mL) and THF (100 mL) at
23.degree. C. After stirring the reaction overnight, the reaction
was adjusted to pH=7. The solvent was evaporated and the residue
was purified by trituration with hexanes and water to afford
N-hydroxy-2-(4-(4-phenylbutyl)phenyl)acetamide 174 in 4.66 g (93%).
(dDMSO) .delta. (ppm) .sup.1H, 10.59 (s, 1H), 8.77 (s, 1H),
7.25-7.06 (m, 9H), 3.19 (s, 2H), 2.58-2.53 (m, 4H), 1.54-1.52 (m,
4H). LRMS: 283.1 (calc) 282.0 (MH).sup.-.
Example 41
N-hydroxy-2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetamide
(179)
Step 1: Methyl 2-(4-(3-hydroxyprop-1-ynyl)phenyl)acetate
##STR00152##
[0772] Following the procedure of Step 2 of Example 40 above
afforded methyl 2-(4-(3-hydroxyprop-1-ynyl)phenyl)acetate in 1.17 g
(79%) as a thick yellow oil. (MeOD-d4) .delta. (ppm) .sup.1H, 7.36
(d, J=8.4 Hz, 2H), 7.24 (d, J=8.0 Hz, 2H), 4.38 (s, 2H), 3.67 (s,
3H), 3.65 (s, 2H).
Step 2: Methyl 2-(4-(3-hydroxypropyl)phenyl)acetate
##STR00153##
[0774] Following the procedure of Step 3 of Example 40 above
afforded methyl 2-(4-(3-hydroxypropyl)phenyl)acetate in 1.18 g
(99%) a clear translucent oil. (MeOD-d4) .delta. (ppm) .sup.1H,
7.15 (d, J=1.6 Hz, 4H), 3.66 (s, 3H), 3.59 (s, 2H), 3.55 (t, J=6.4
Hz, 2H), 2.65 (t, J=7.6 Hz, 2H), 1.82 (m, 2H).
Step 3: Methyl 2-(4-(3-oxopropyl)phenyl)acetate
##STR00154##
[0776] TEMPO (0.02 equiv, 6 mg, 0.038 mmol) was added to a solution
of methyl 2-(4-(3-hydroxypropyl)phenyl)acetate in CH.sub.2Cl.sub.2
(5 mL) and cooled to 0.degree. C. KBr (2.2 equiv, 1.6 mL, 2.7M) and
KHCO.sub.3 (5.5 equiv, 6.6 mL, 1.6M) solutions were then added
followed by the dropwise addition of 10% aq. NaOCl (1.34 equiv, 1.9
g, 2.6 mmol). Once the addition was complete, the reaction was
stirred for an additional 10 min. at 0.degree. C. The reaction was
quenched with sat. aq.sodium thiosulfate solution (3 mL) and then
partitioned between H.sub.2O (10 mL) and CH.sub.2Cl.sub.2 (10 mL).
The organic phase was separated and the remaining aqueous phase was
extracted with EtOAc (2.times.2 mL). The organic layers were
combined, dried over Na.sub.2SO.sub.4, filtered and concentrated
under reduced pressure. The resulting aldehyde was carried forward
to the subsequent reaction without further purification. LRMS:
206.1 (calc) 207.2 (found) (MH).sup.+.
Step 4: Methyl 2-(4-(but-3-ynyl)phenyl)acetate
##STR00155##
[0778] Dimethyl-2-oxopropylphosphonate (1.2 equiv, 1.5 g, 9.1 mmol)
was added to a suspension of K.sub.2CO.sub.3 (3 equiv, 3.1 g, 23
mmol) and p-TsN.sub.3 (1.2 equiv, 1.8 g, 9.1 mmol) in MeCN (144
mL). The mixture was then stirred for 2 h. A solution of the methyl
2-(4-(3-oxopropyl)phenyl)acetate (1 equiv, 1.56 g, 7 6 mmol) in
MeOH was then added in one portion and the reaction was stirred
overnight. The solvent was evaporated under reduced pressure. The
residue was partitioned between Et.sub.2O (100 mL) and water (100
mL). The aqueous phase was separated and extracted with Et.sub.2O
(25 mL) twice. The organic phases were combined, dried over
Na.sub.2SO.sub.4, filtered and concentrated. The residue was
purified by silica gel flash column eluting with 0-30% EtOAc in
hexanes to afford methyl 2-(4-(but-3-ynyl)phenyl)acetate in 980 mg
(64%). LRMS: 202.5 (calc) 225.1 (found) (MNa).sup.+.
Step 5: Methyl
2-(4-(4-(2,4,5-trifluorophenyl)but-3-ynyl)phenyl)acetate
##STR00156##
[0780] Following the procedure of Step 2 of Example 40 above
afforded methyl
2-(4-(4-(2,4,5-trifluorophenyl)but-3-ynyl)phenyl)acetate in 95 mg
(39%) as a yellow oil. LRMS: 332.3 (calc) 355.3 (found)
(MNa).sup.+.
Step 6: Methyl
2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetate
##STR00157##
[0782] Following the procedure of Step 3 of Example 40 above
afforded methyl 2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetate
in 91 mg (95%) as a clear translucent oil. LRMS: 336.3 (calc) 359.2
(found) (MNa).sup.+.
Step 7:
N-hydroxy-2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetamide
(179)
##STR00158##
[0784] Following the procedure of Step 4 of Example 40 above
afforded
N-hydroxy-2-(4-(4-(2,4,5-trifluorophenyl)butyl)phenyl)acetamide 179
in 52 mg (57%) as a white powder. (MeOD-d4) .delta. (ppm) .sup.1H,
7.15 (m, 6H), 3.35 (s, 2H), 2.61 (m, 4H), 1.60 (m, 4H). LRMS: 337.3
(calc) 338.3 (found) (MH).sup.+.
Example 42
2-(4-(4-(benzo[c][1,2,5]oxadiazol-5-yl)but-3-ynyl)phenyl)-N-hydroxyacetami-
de (180)
##STR00159##
[0786] Following the procedure of Step 4 of Example 40 above
afforded
2-(4-(4-(benzo[c][1,2,5]oxadiazol-5-yl)but-3-ynyl)phenyl)-N-hydroxyacetam-
ide 180 in 8 mg (57%) as a yellowish orange powder. (MeOD-d4)
.delta. (ppm) .sup.1H, 10.62 (s, 1H), 8.79 (s, 1H), 8.07 (s, 1H),
8.03 (d, J=9.2 Hz, 1H), 7.40 (d, J=9.6 Hz, 1H), 7.24 (d, J=8.4 Hz,
2H), 7.18 (d, J=8.0 hz, 2H), 3.23 (s, 3H), 2.83 (m, 2H), 2.77 (m,
2H) LRMS: 321.1 (calc) 320.3 (found) (MH).sup.-.
Example 43
N-Hydroxy-2-(3-(4-phenylbutyl)phenyl)acetamide (181)
Step 1: Methyl 2-(3-(4-hydroxybut-1-ynyl)phenyl)acetate
##STR00160##
[0788] Following the procedure of Step 2 of Example 40 above
afforded methyl 2-(3-(4-hydroxybut-1-ynyl)phenyl)acetate in 227 mg
(34%) as a yellow oil. LRMS: 218.2 (calc) 219.1 (found)
(MH).sup.+.
Step 2: Methyl 2-(3-(4-hydroxybutyl)phenyl)acetate
##STR00161##
[0790] Following the procedure of Step 3 of Example 40 above
afforded methyl 2-(3-(4-hydroxybutyl)phenyl)acetate in 181 mg (78%)
as a clear translucent oil. LRMS: 222.3 (calc) 223.1 (found)
(MH).sup.+.
Step 3: Methyl 2-(3-(4-oxobutyl)phenyl)acetate
##STR00162##
[0792] Following the procedure of Step 3 of Example 41 above
afforded methyl 2-(3-(4-oxobutyl)phenyl)acetate in 178 mg (99%) as
a clear translucent oil. LRMS: 220.3 (calc) 221.4 (found)
(MH).sup.+.
Step 4: Methyl 2-(3-(4-hydroxy-4-phenylbutyl)phenyl)acetate
##STR00163##
[0794] A 1.0 M solution of PhMgBr (1 equiv, 0.45 mmol) was added
dropwise to a solution of methyl 2-(3-(4-oxobutyl)phenyl)acetate (1
equiv, 100 mg, 0.45 mmol) in THF (2 mL) at 0.degree. C. The
reaction was then allowed to warm to 23.degree. C. over 1 h. The
reaction was quenched by the addition on sat. aq. NH.sub.4Cl (10
mL). The aqueous phase was separated and extracted with EtOAc
(2.times.5 mL). The organic layers were then combined, dried over
Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure.
The crude product was purified by silica gel column chromatography
eluting with 0-50% EtOAc in hexanes to afford methyl
2-(3-(4-hydroxy-4-phenylbutyl)phenyl)acetate in 78 mg(58%) as a
clear translucent oil. LRMS: 298.4 (calc) 321.2 (found)
(MH).sup.+.
Step 5: Methyl 2-(3-(4-phenylbutyl)phenyl)acetate
##STR00164##
[0796] Following the procedure of Step 3 of Example 40 above
afforded methyl 2-(3-(4-phenylbutyl)phenyl)acetate in 15 mg (20%)
as a clear translucent oil. LRMS: 282.4 (calc) 283.2 (found)
(MH).sup.+.
Step 6: N-Hydroxy-2-(3-(4-phenylbutyl)phenyl)acetamide (181)
##STR00165##
[0798] Following the procedure of Step 4 of Example 40 above
afforded N-Hydroxy-2-(3-(4-phenylbutyl)phenyl)acetamide (181) in 5
mg (33%) as a white powder. (CD.sub.3OD) .delta. (ppm) 1H, 7.23 (m,
9H), 3.36 (s, 2H), 2.61 (m, 4H), 1.63 (m, 4H) LRMS (ESI): (calc.)
283.1 (found) 282.2 (MH).sup.-
Example 44
[0799] The following additional compounds were prepared by
procedures analogous to those described in the Examples 40-43:
[0800] a)
N-hydroxy-2-(4-(4-(4-(trifluoromethyl)phenyl)butyl)phenyl)acetamide
(182)
##STR00166##
[0801] (CD3OD) .delta. (ppm) 1H, 7.53 (d, J=8.4 Hz, 2H), 7.33 (d,
J=8.0 Hz, 2H), 7.18 (d, J=8.0 Hz, 2H), 7.10 (d, J=8.0 Hz, 2H), (s,
2H), 2.70 (t, J=7.2 Hz, 2H), 2.61 (t, J=7.2 Hz, 2H), 1.63 (m, 4H).
LRMS (ESI): (calc.) 351.14 (found) 350.24 (MH).sup.-. [0802] b)
2-(4-(4-(1H-indol-5-yl)butyl)phenyl)-N-hydroxyacetamide (183)
##STR00167##
[0803] (CD3OD) d(ppm) 1H, 7.29 (m, 1H), 7.24 (d, J=8.8 Hz, 1H),
7.18-7.16 (m, 3H), 7.11-7.09 (m, 2H), 6.90 (dd, J=8.4 Hz, 1.6 Hz,
1H), 6.34 (d, J=4 Hz, 1H), 3.35 (s, 2H), 2.68 (t, J=7.2 Hz, 2H),
2.61 (t, J=7.2 Hz, 2H), 1.64 (m, 4H) LRMS (ESI): (calc.) 322.17
(found) 323.421 (MH).sup.+. [0804] c)
N-hydroxy-2-(4-(4-(3-(trifluoromethyl)phenyl)butyl)phenyl)aceta-
mide (184)
##STR00168##
[0805] (dDMSO) .delta. (ppm) 1H, 10.60 (s, 1H), 8.77 (s, 1H), 7.50
(m, 4H), 7.12 (d, J=8.0 Hz, 2H), 7.07 (d, J=8.0 Hz, 2H), 3.19 (s,
2H), 2.68 (t, J=7.2 Hz, 2H), 2.54 (t, J=7.2 Hz, 2H), 1.55 (m, 4H).
LRMS (ESI): (calc.) 351.1 (found) 350.3 (MH).sup.-. [0806] d)
N-hydroxy-2-(4-(4-(2-(trifluoromethyl)phenyl)butyl)phenyl)acetamide
(185)
##STR00169##
[0807] (dDMSO) d(ppm) 1H, 10.60 (s, 1H), 8.77 (s, 1H), 7.63 (d,
J=8.0 Hz, 1H), 7.57 (t, J=7.2 Hz, 1H), 7.43 (d, J=7.6 Hz, 1H), 7.37
(t, J=8.0 Hz, 1H), 7.13 (d, J=8.0 Hz, 2H), 7.09 d, J=8.0 Hz, 2H),
3.20 (s, 2H), 2.73 (t, J=7.6 Hz, 2H), 1.58 (m, 4H). LRMS (ESI):
(calc.) 351.1 (found) 350.3 (MH)-. [0808] e)
N-hydroxy-2-(4-(4-(imidazo[1,2-a]pyridin-6-yl)butyl)phenyl)acetamide
(186)
##STR00170##
[0809] (CD3OD) d(ppm) 1H, 8.19 (s, 1H), 7.73 (s, 1H), 7.49 (s, 1H),
7.43 (d, J=9.2 Hz, 1H), 7.15 (m, 5H), 3.34 (s, 2H), 2.63 (m, 4H),
1.66 (m, 4H). LRMS (ESI): (calc.) 323.1 (found) 324.3 (MH)+ [0810]
f)
2-(4-(4-(benzo[d][1,3]dioxol-5-yl)butyl)phenyl)-N-hydroxyacetamide
(187)
##STR00171##
[0811] (dDMSO) d(ppm) 1H, 10.60 (s, 1H), 8.78 (s, 1H), 7.12 (d,
J=8.0 Hz, 2H), 7.07 (d, J=8.0 Hz, 2H), 6.75 (m, 2H), 6.60 (d, J=8.0
Hz, 1H), 5.92 (s, 2H), 3.19 (s, 2H), 2.52 (m, 4H), 1.50 (m, 4H).
LRMS (ESI): (calc.) 327.1 (found) 326.4 (MH)-. [0812] g)
2-(4-(4-(benzo[d][1,3]dioxol-5-yl)but-3-ynyl)phenyl)-N-hydroxyacetamide
(188)
##STR00172##
[0813] (dDMSO) d(ppm) 1H, 10.61 (s, 1H), 8.78 (s, 1H), 7.20 (d,
J=8.4 Hz, 2H), 7.16 (d, J=8.4 Hz, 2H), 6.85 (m, 3H), 6.01 (s, 2H),
3.22 (s, 2H), 2.77 (t, J=7.6 Hz, 2H), 2.61 (t, J=6.8 Hz, 2H). LRMS
(ESI): (calc.) 323.1 (found) 322.2 (MH)-. [0814] h)
2-(4-(4-(benzo[c][1,2,5]oxadiazol-5-yl)but-3-ynyl)phenyl)-N-hydroxyacetam-
ide (189)
##STR00173##
[0815] (dDMSO) d(ppm) 1H, 10.60 (s, 1H), 8.78 (s, 1H), 7.12 (d,
J=8.0 Hz, 2H), 7.07 (d, J=8.0 Hz, 2H), 6.75 (m, 2H), 6.60 (d, J=8.0
Hz, 1H), 5.92 (s, 2H), 3.19 (s, 2H), 2.52 (m, 4H), 1.50 (m, 4H).
LRMS (ESI): (calc.) 327.1 (found) 326.4 (MH)-. [0816] i)
N-hydroxy-2-(4-(2-phenoxyethoxy)phenyl)acetamide (190)
##STR00174##
[0817] (DMSO) d(ppm) 1H, 10.58 (bs, 1H), 8.77 (bs, 1H), 7.28 (t,
J=7.6 Hz<2H), 7.15 (d, J=8.8 Hz, 2H), 6.93 (m, 5H), 4.26 (s,
4H), 3.18 (s, 2H). LRMS (ESI): (calc.) 287.3 (found) 310.0
(MH).sup.+. [0818] j)
N-hydroxy-2-(4-(3-phenoxypropyl)phenyl)acetamide (191)
##STR00175##
[0819] (CD3OD) d(ppm) 1H, 7.21 (m, 6H), 6.88 (m, 6H), 3.92 (t, J=8
Hz, 2H), 3.35 (s, 2H), 2.77 (t, J=7.2 Hz, 2H), 2.04 (m, 2H). LRMS
(ESI): (calc.) 285.3 (found) 286.2 (MH).sup.+. [0820] k)
2-(4-butylphenyl)-N-hydroxyacetamide (192)
##STR00176##
[0821] (CD3OD) d(ppm) 1H, 7.18 (d, J=8 Hz, 2H), 7.11 (d, J=8 Hz,
2H), 3.35 (s, 2H), 2.57 (T, J=7.6 Hz, 2H), 1.56 (m, 2H), 1.33 (m,
2H), 0.92 (t, J=7.2 Hz, 2H). LRMS (ESI): (calc.) 207.2 (found)
208.1 (ES+;Na+). [0822] l)
N-hydroxy-2-(4-(5-phenylpentyl)phenyl)acetamide (193)
##STR00177##
[0823] (CD3OD) d(ppm) 1H, 7.22-7.08 (m, 9H), 3.34 (s, 2H), 2.56 (m,
4H), 1.61 (m, 4H), 1.34 (m, 2H). LRMS (ESI): (calc.) 297.4 (found)
298.3 (MH).sup.+. [0824] m)
N-hydroxy-2-(4-(4-hydroxy-4-(pyridin-4-yl)butyl)phenyl)acetamide
(194)
##STR00178##
[0825] (CD3OD) d(ppm) 1H, 8.44 (d, J=6.0 Hz, 2H), 7.36 (d, J=6.4
Hz, 2H), 7.18 (d, J=8.4 Hz, 2H), 7.09 (d, J=8.0 Hz, 2H), 4.65 (m,
1H), 3.34 s, 2H), 2.60 (m, 2H), 1.69 (m, 4H). LRMS (ESI): (calc.)
300.3 (found) 301.3 (MH).sup.+. [0826] n)
N-hydroxy-2-(4-(4-hydroxy-4-(pyridin-3-yl)butyl)phenyl)acetamide
(195)
##STR00179##
[0827] (CD3OD) d(ppm) 1H, 8.46 (s, 1H), 8.41 (d, J=4.8 Hz, 1H),
7.78 (d, J=8.0 Hz, 1H), 7.39 (t, J=5.6 Hz, 1H), 7.18 (d, J=8.0 Hz,
2H), 7.09 (d, J=8.0 Hz, 2H), 4.69 (m, 1H), 3.34 (s, 2H), 2.61 ((t,
J=6.4 Hz, 2H), 1.79-1.58 (m, 4H). LRMS (ESI): (calc.) 300.5 (found)
301.4 (MH)+. [0828] o)
N-hydroxy-2-(4-(4-hydroxy-4-(pyridin-2-yl)butyl)phenyl)acetamide
(196)
##STR00180##
[0829] (CD3OD) d(ppm) 1H, 8.42 (m, 1H), 7.81 (m, 1H), 7.51 (d,
J=7.2 Hz, 1H), 7.26 (d, J=4.8 Hz, 1H), 7.16 (m, 1H), 7.08 (m, 2H),
4.69 (m, 1H), 3.34 (s, 2H), 2.59 (m, 2H), 1.76-1.69 (m, 4H). LRMS
(ESI): (calc.) 300.3 (found) 301.4 (MH).sup.+. [0830] p)
N-hydroxy-2-(4-(4-(pyridin-4-yl)butyl)phenyl)acetamide (197)
##STR00181##
[0831] (CD3OD) d(ppm) 1H, 8.37 (d, J=6.0 Hz, 2H), 7.24 (d, J=5.6
Hz, 2H), 7.18 (d, J=8.0 Hz, 2H), 7.10 (d, J=8.0 Hz, 2H), 3.35 (s,
2H), 2.67 (t, J=6.4 Hz, 2H), 2.62 (t, J=7.2 Hz, 2H), 1.65 (m, 4H).
LRMS (ESI): (calc.) 284.1 (found) 285.3 (MH)+. [0832] q)
N-hydroxy-2-(4-(4-(pyridin-3-yl)butyl)phenyl)acetamide (198)
##STR00182##
[0833] (CD3OD) d(ppm) 1H, 8.24 (m, 2H), 7.65 (d, J=8.0 Hz, 1H),
7.33 (m, 1H), 7.18 (d, J=7.6 Hz, 2H), 7.10 (d, J=7.6 Hz, 2H), 3.35
(s, 2H), 2.64 (m, 4H), 1.64 (m, 4H). LRMS (ESI): (calc.) 284.1
(found) 285.3 (MH)+. [0834] r)
N-hydroxy-2-(4-(4-(pyridin-2-yl)butyl)phenyl)acetamide (199)
##STR00183##
[0835] (CD3OD) d(ppm) 1H, 8.39 (d, J=6.0 Hz), 7.75 (t, J=8.0 Hz,
1H), 7.23 (m, 4H), 7.10 (d, J=8.0 Hz, 2H), 3.34 (s, 2H), 2.78 (t,
J=7.6 Hz, 2H), 2.61 (t, J=7.6 Hz, 2H), 1.67 (m, 4H). LRMS (ESI):
(calc.) 284.1 (found) 285.4 (MH)+. [0836] s)
N-hydroxy-2-(4-(3-phenylpropyl)phenyl)acetamide (200)
##STR00184##
[0837] (CD3OD) d(ppm): 7.24-7.11 (m, 9H), 3.35 (s, 2H), 2.60 (t,
4H, J=7.6 Hz), 1.89 (m, 2H). LRMS: 269.1 (calc) 270.1 (found).
[0838] t) N-hydroxy-4-(5-phenylpentyl)benzamide (201)
##STR00185##
[0839] (CD3OD) d(ppm) 1H, 7.64 (d, J=7.6 Hz, 2H), 7.22 (m, 4H),
7.12 (m, 3H), 2.63 (t, J=7.6 Hz, 2H), 2.57 (t, J=7.2 Hz, 2H), 1.63
(m, 4H), 1.35 (m, 2H). LRMS (ESI): (calc.) 283.4 (found) 284.3
(MH)+.
Example 45
N-(1-aminocyclopropanecarbonyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide
hydrochloride (174a)
Step 1:
tert-butyl-1-1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)-
cyclopropylcarbamate
##STR00186##
[0841] N-hydroxy-2-(4-(4-phenylbutyl)phenyl)acetamide 174 (1 equiv,
156 mg, 0.55 mmol) was dissolved in DMF (3 mL).
1-(tert-butoxycarbonylamino)cyclopropanecarboxylic acid (1.5 equiv,
166 mg, 0.83 mmol) was then added followed by the sequential
addition of HOBt (1 equiv, 74 mg, 0.55 mmol) and EDC (1.5 equiv,
158 mg, 0.83 mmol). The reaction was then stirred overnight. The
reaction was then partitioned between EtOAc (5 mL) and H.sub.2O (5
mL). The organic phase was separated, dried over Na.sub.2SO.sub.4,
filtered and concentrated. The residue was purified by trituration
with Et.sub.2O to afford
tert-butyl-1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)cycloprop-
ylcarbamate in 166 mg (65%). LRMS: 466.5 (calc) 489.3
(MNa).sup.+.
Step 2:
N-(1-aminocyclopropanecarbonyloxy)-2-(4-(4-phenylbutyl)phenyl)acet-
amide hydrochloride (174a)
##STR00187##
[0843] A 4M solution of HCl in dioxane (3 mL) was added to
tert-butyl-1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)cycloprop-
ylcarbamate (166 mg, 0.36 mmol). The reaction was stirred for 1 h.
The solvent was evaporated under reduced pressure. The compound was
triturated with Et.sub.2O to afford
N-(1-aminocyclopropanecarbonyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide
hydrochloride (174a) in 142 mg (99%). (dDMSO) .delta. (ppm)
.sup.1H, 12.36 (bs, 1H), 8.82 (bs, 3H), 7.26-7.08 (m, 9H), 3.43 (s,
2H), 2.55 (m, 4H), 1.55-1.49 (m, 8H). LRMS: 366.1 (calc) 365.4
(MH).sup.-.
Example 46
[0844] The following additional compounds were prepared by
procedures analogous to those described in the Examples 40-43 and
45: [0845] a)
(S)--N-(2-amino-3-phenylpropanoyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamid-
e hydrochloride (174b)
##STR00188##
[0846] (DMSO) .delta. (ppm) 1H, 12.39 (bs, 1H), 8.47 (bs, 3H),
7.31-6.96 (m, 1H), 4.54 (m, 1H), 3.46 (s, 2H), 3.15 (m, 2H), 2.56
(q, J=6.8, 13.6 Hz, 4H), 1.54 (m, 4H). LRMS (ESI): (calc.) 430.2
(found) 431.4 (MH).sup.+. [0847] b)
(S)--N-(2-amino-3-methylbutanoyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide
hydrochloride (174c)
##STR00189##
[0848] (DMSO) .delta. (ppm) 1H, 12.32 (bs, 1H), 8.33 (bs, 3H),
7.24-7.09 (m, 9H), 4.13 (m, 1H), 3.45 (s, 2H), 2.57 (m, 4H), 2.18
(m, 1H), 1.55 (m, 4H), 1.03-0.94 (m, 6H). LRMS (ESI): (calc.) 382.2
(found) 383.1 (MH).sup.+. [0849] c)
(S)--N-(2-amino-3,3-dimethylbutanoyloxy)-2-(4-(4-phenylbutyl)phenyl)aceta-
mide hydrochloride (174d)
##STR00190##
[0850] (dDMSO) .delta. (ppm) 1H, 12.49 (bs, 1H), 8.59 (bs, 3H),
7.26-7.09 (m, 9H), 3.98 (s, 1H), 3.47 (s, 2H), 2.56 (m, 4H), 1.55
(m, 4H), 1.05 (s, 9H). LRMS (ESI): (calc.) 396.2 (found) 397.5
(MH).sup.+. [0851] d)
N-(1-aminocyclobutanecarbonyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide
hydrochloride (174e)
##STR00191##
[0852] (dDMSO) .delta. (ppm) 1H, 12.52 (bs, 1H), 8.90 (bs, 1H),
7.19 (m, 9H), 3.47 (s, 2H), 2.56 (m, 6H), 2.05 (m, 2H), 1.54 (m,
4H). LRMS (ESI): (calc.) 380.2 (found) 381.4 (MH).sup.+. [0853] e)
N-(2-amino-2-methylpropanoyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide
hydrochloride (174f)
##STR00192##
[0854] (dDMSO) .delta. (ppm) 1H, 12.45 (bs, 1H), 8.77 (bs, 3H),
7.16 (m, 9H), 3.45 (s, 2H), 2.56 (m, 4H), 1.54 (m, 10H). LRMS
(ESI): (calc.) 368.2 (found) 369.4 (MH).sup.+. [0855] f)
N-(1-(aminomethyl)cyclopropanecarbonyloxy)-2-(4-(4-phenylbutyl)phenyl)ace-
tamide hydrochloride (174g)
##STR00193##
[0856] (dDMSO) .delta. (ppm) 1H, 12.17 (bs, 1H), 8.00 (bs, 3H),
7.26-7.08 (m, 9H), 3.42 (s, 2H), 3.05 (s, 2H), 2.55 (m, 4H), 1.54
(m, 4H), 1.35 (m, 2H), 1.26 (m, 2H) LRMS (ESI): (calc.) 380.2
(found) 381.5 (MH).sup.+
Example 47
[0857] The following additional prodrugs according to the present
invention were also prepared. [0858]
(S)--N-(2,6-diaminohexanoyloxyl-2-(4-(4-phenylbutyl)phenyl)acetamide
##STR00194##
[0859] LRMS (ESI): (calc.) 411.25 (found) 412.509 (MH)+
[0860] (DMSO) d(ppm) 1H, 12.50 (s, 1H), 8.66 (s, 3H), 7.81 (s, 3H),
7.27-7.10 (m, 9H), 4.27-4.21 (m, 1H), 3.48-3.46 (m, 2H), 2.75-2.68
(m, 2H), 2.60-2.52 (m, 4H), 1.88-1.80 (m, 2H), 1.60-1.40 (m, 8H).
[0861]
N-(2-hydroxyacetoxy)-2-(4-(4-phenylbutyl)phenyl)acetamide
##STR00195##
[0862] LRMS (ES-): (calc.) 341.16 (found) 340.466 (MH)+
[0863] (DMSO) d(ppm) 1H, 11.95 (s, 1H), 7.27-7.09 (m, 9H), 5.64 (t,
J=6.4 Hz, 1H), 4.17 (d, J=6.4 Hz, 2H), 3.42 (s, 2H), 2.60-2.52 (m,
4H), 1.58-1.53 (m, 4H) [0864]
(S)--N-(2-amino-5-guanidinopentanoyloxy)-2-(4-(4-phenylbutyl)phenyl)aceta-
mide
##STR00196##
[0865] LRMS (ESI): (calc.) 439.26 (found) 440.651 (MH)+
[0866] (DMSO) d(ppm) 1H, 12.60 (s, 1H), 8.77 (s, 1H), 8.43 (s, 4H),
8.00 (s, 1H), 7.85 (s, 1H), 7.37-7.12 (m, 9H), 3.95-3.85 (m, 1H),
3.20-3.10 (m, 4H), 2.60-2.52 (m, 2H), 1.92-1.72 (m, 4H), 1.70-1.45
(m, 4H) [0867]
(S)-2,6-diamino-N-(1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)c-
yclopropyl)hexanamide
##STR00197##
[0868] LRMS (ESI): (calc.) 494.29 (found) 495.592 (MH)+
[0869] (DMSO) d(ppm) 1H, 12.08 (s, 1H), 9.36 (s, 1H), 8.23 (s, 3H),
7.86 (s, 3H), 7.27-7.08 (m, 9H), 3.75-3.65 (m, 1H), 3.39 (s, 2H),
2.78-2.67 (m, 2H), 2.62-2.52 (m, 4H), 1.80-1.66 (m, 2H), 1.64-1.46
(m, 8H), 1.44-1.28 (m, 2H), 1.38-1.26 (m, 2H) [0870]
N-(1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbonyl)cyclopropyl)nicot-
inamide
##STR00198##
[0871] LRMS (ESI): (calc.) 471.22 (found) 472.525 (MH)+
[0872] (DMSO) d(ppm) 1H, 9.45 (s, 1H), 9.00 (s, 1H), 8.72 (d, 1H,
J=4.8 Hz), 8.19 (d, 1H, J=8 Hz), 7.51 (dd, 1H, J=8 Hz, 4.8 Hz),
7.27-7.05 (m, 9H), 2.59-2.53 (m, 4H), 1.60-1.50 (m, 4H), 1.32-1.27
(m, 2H), 1.25-1.21 (m, 2H) [0873]
(S)-2-amino-3-methyl-N-(1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbo-
nyl)cyclopropyl)butanamide
##STR00199##
[0874] LRMS (ESI): (calc.) 465.26 (found) 466.695 (MH)+
[0875] (DMSO) d(ppm) 1H, 12.04 (s, 1H), 9.21 (s, 1H), 8.09 (s, 3H),
7.27-7.06 (m, 9H), 3.50-3.45 (m, 1H), 3.40-3.37 (m, 2H), 2.61-2.53
(m, 4H), 2.14-2.04 (m, 1H), 1.62-1.48 (m, 6H), 1.24-1.16 (m, 2H),
0.98-0.90 (m, 6H) [0876]
(S)-2-amino-3-phenyl-N-(1-((2-(4-(4-phenylbutyl)phenyl)acetamidooxy)carbo-
nyl)cyclopropyl)propanamide
##STR00200##
[0877] LRMS (ESI): (calc.) 513.26 (found) 514.675 (MH)+
[0878] (DMSO) d(ppm) 1H, 12.03 (s, 1H), 9.19 (s, 1H), 8.14 (s, 3H),
7.29-7.08 (m, 14H), 3.92-3.85 (m, 1H), 3.40-3.37 (m, 2H), 3.06-2.94
(m, 2H), 2.60-2.53 (m, 4H), 1.57-1.51 (m, 4H), 1.51-1.46 (m, 2H),
1.40-0.80 (m, 2H) [0879]
2-(4-(4-phenylbutyl)phenyl)-N-((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydro-
xymethyl)tetrahydro-2H-pyran-2-yloxy)acetamide
##STR00201##
[0880] LRMS (ESI): (calc.) 445.2 (found) 446.6 (MH)+
[0881] (CD3OD) d(ppm) 1H, 7.24-7.10 (m, 9H), 4.50 (d, J=8.0 Hz,
1H), 3.90 (dd, J=2.4, 12.0 Hz, 1H), 3.65 (dd, J=6.0, 11.6 Hz, 1H),
3.40 (s, 2H), 3.35-3.25 (m, 3H), 2.61 (m, 4H), 1.62 (m, 4H) [0882]
N-(2,3-dihydroxypropanoyloxy)-2-(4-(4-phenylbutyl)phenyl)acetamide
##STR00202##
[0883] LRMS (ESI): (calc.) 371.17 (found) 000.0 (M)-370.478
[0884] (CD3OD) d(ppm) 1H, 7.25-7.08 (m, 9H), 4.41 (t, J=4 HZ, 1H),
3.81 (d, J=4 Hz, 2H), 3.51 (s, 2H), 2.64-2.58 (m, 4H), 1.65-1.58
(m, 4H) [0885]
N-hydroxy-2-(4-(4-(3-morpholinophenyl)butyl)phenyl)acetamide
##STR00203##
[0886] LRMS (ESI): (calc.) 368.21 (found) 369.571 (MH)+
[0887] (dmso) d(ppm) 1H, 10.62 (s, 1H), 8.78 (s, 1H), 7.14-7.04 (m,
5H), 6.72-6.68 (m, 2H), 6.60 (d, J=7.2 Hz, 1H), 3.70 (t, J=4.8 Hz,
4H), 3.20 (s, 2H), 3.04 (t, J=4.8 Hz, 4H), 2.56-2.49 (m, 4H),
1.58-1.48 (m, 4H) [0888]
N-hydroxy-2-phenyl-2-(4-(4-phenylbutyl)phenyl)acetamide
##STR00204##
[0889] LRMS (ESI): (calc.) 359.2 (found) 360.4 (MH)+
[0890] (CD3OD) d(ppm) 1H, 7.30-7.11 (m, 14H), 4.73 (s, 1H), 2.60
(m, 4H), 1.61 (m, 4H)
##STR00205##
[0891] LRMS (ESI): (calc.) 431.5 (found) 431.5 (M)+
[0892] (CD3OD) d(ppm) 1H, 8.84 (s, 1H), 8.71 (d, J=6.0 Hz, 1H),
8.49 (d, J=8.0 Hz, 1H), 7.96 (t, J=6.8 Hz, 1H), 7.24-7.10 (m, 9H),
4.30 (s, 3H), 3.47 (s, 2H), 3.22 (t, J=7.2 Hz, 2H), 2.97 (t, J=6.8
Hz, 2H), 2.61 (m, 4H), 1.62 (m, 4H) [0893]
2-(4-(4-(fluorophenyl)butyl)phenyl)-N-hydroxyamide
##STR00206##
[0894] LRMS (ESI): (calc.) 301.1 (found) 302.3 (MH)+
[0895] (dDMSO) d(ppm) 1H, 7.19 (m, 2H), 7.11-6.98 (m, 6H), 3.02 (s,
2H), 2.54 (m, 4H), 1.53 (m, 4H)
##STR00207##
[0896] LRMS (ESI): (calc.) 421.2 (found) 421.5 (MH)+
[0897] (CD3OD) d(ppm) 1H, 12.13 (bs, 1H), 10.11 (s, 1H), 9.14 (s,
1H), 7.27-7.09 (m, 9H), 4.67 (t, J=6.0 Hz, 2H), 3.86 (s, 3H), 3.39
(s, 2H), 3.17 (t, J=6.0 Hz, 2H), 2.56 (m, 4H), 1.55 (m, 4H)
##STR00208##
[0898] LRMS (ESI): (calc.) 403.2 (found) 403.4 (M)+.
[0899] (CD3OD) d(ppm) 1H, 9.66 (s, 1), 9.14 (m, 2), 8.25 (m, 1H),
7.26-7.10 (m, 9H), 4.49 (s, 3H), 3.60 (s, 2H), 2.62 (m, 4H), 1.64
(m, 4H)
Example 48
Inhibition of Histone Deacetylase Enzymatic Activity
[0900] HDAC inhibitors are screened against histone deacetylase
enzyme in nuclear extracts prepared from the human small cell lung
cancer cell line H446 (ATTC HTB-171) and against a cloned
recombinant human (e.g., HDAC-1) enzyme expressed and purified from
a Baculovirus insect cell expression system.
[0901] For deacetylase assays, 20,000 cpm of the
[.sup.3H]-metabolically labeled acetylated histone substrate (M.
Yoshida et al., J. Biol. Chem. 265(28): 17174-17179 (1990)) is
incubated with 30 mg of H446 nuclear extract or an equivalent
amount of the cloned recombinant hHDAC-1 for 10 minutes at
37.degree. C. The reaction is stopped by adding acetic acid (0.04
M, final concentration) and HCl (250 mM, final concentration). The
mixture is extracted with ethyl acetate and the released
[.sup.3H]-acetic acid was quantified by scintillation counting. For
inhibition studies, the enzyme is preincubated with compounds at
4.degree. C. for 30 minutes prior to initiation of the enzymatic
assay. IC.sub.50 values for HDAC enzyme inhibitors are determined
by performing dose response curves with individual compounds and
determining the concentration of inhibitor producing fifty percent
of the maximal inhibition.
[0902] Alternatively (Table 4a), the following protocol is used to
assay the compounds of the invention. In the assay, the buffer used
is 25 mM HEPES, pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl.sub.2
and the substrate is Boc-Lys(Ac)-AMC in a 50 mM stock solution in
DMSO. The enzyme stock solution is 4.08 .mu.g/mL in buffer. The
compounds are pre-incubated (2 .mu.l in DMSO diluted to 13 .mu.l in
buffer for transfer to assay plate) with enzyme (20 .mu.l of 4.08
.mu.g/ml) for 10 minutes at room temperature (35 .mu.l
pre-incubation volume). The mixture is pre-incubated for 5 minutes
at room temperature. The reaction is started by bringing the
temperature to 37.degree. C. and adding 16 .mu.l substrate. Total
reaction volume is 50 .mu.l. The reaction is stopped after 20
minutes by addition of 50 .mu.l developer, prepared as directed by
Biomol (Fluor-de-Lys developer, Cat. #KI-105). A plate is incubated
in the dark for 10 minutes at room temperature before reading
(.lamda..sub.Ex=360 nm, .lamda..sub.Em=470 nm, Cutoff filter at 435
nm).
[0903] Representative data are presented in Tables 4 and 4a. In the
first column of Table 4 are reported IC.sub.50 values determined
against histone deacetylase in nuclear extracts from H446 cells
(pooled HDACs). In the second column of Table 4 are reported
IC.sub.50 values determined against recombinant human HDAC-1 enzyme
(rHDAC-1). For less active compounds, the data are expressed as the
percent inhibition at the specified concentration.
TABLE-US-00005 TABLE 4 Inhibition of Histone Deacetylase pooled
HDACs rHDAC-1 Example Cpd. Structure IC.sub.50 (.mu.M) IC.sub.50
(.mu.M) Ex. 1 4 ##STR00209## 7 Ex. 2 7 ##STR00210## 70 Ex. 3 8
##STR00211## 15 Ex. 4 9 ##STR00212## 9 Ex. 5 10 ##STR00213## 30 Ex.
6 11 ##STR00214## 10 Ex. 7 12 ##STR00215## 3 Ex. 8 13 ##STR00216##
0.9 Ex. 9 14 ##STR00217## 36% @ 100 .mu.M Ex. 10 15 ##STR00218## 25
Ex. 11 16 ##STR00219## 38% 100 .mu.M Ex. 12 17 ##STR00220## 47% 100
.mu.M Ex. 13 18 ##STR00221## 160 Ex. 14 19 ##STR00222## 20 Ex. 15
26 ##STR00223## <20 Ex. 16 32 ##STR00224## <20 Ex. 17 34
##STR00225## 2 0.3 Ex. 18 36 ##STR00226## 0.5 0.2 Ex. 19 38
##STR00227## 0.75 0.1 Ex. 20 42 ##STR00228## 5 1.0 Ex. 21 45
##STR00229## 4 Ex. 22 50 ##STR00230## 5 Ex. 23 53 ##STR00231## 25
Ex. 24 56 ##STR00232## 15 Ex. 25 61 ##STR00233## 4 Ex. 26 64
##STR00234## 12% @ 100 .mu.M Ex. 27 68 ##STR00235## 3% @ 20 .mu.M
Ex. 28 70 ##STR00236## 5.5 0.9 Ex. 28 71 ##STR00237## 44% @ 20
.mu.M Ex. 28 73 ##STR00238## 35% @ 20 .mu.M Ex. 29 77 ##STR00239##
.quadrature. 0.65 Ex. 30 81 ##STR00240## >50 >25 Ex. 31 86
##STR00241## 3.8 Ex. 31 87 ##STR00242## 3 0.6 Ex. 31 88
##STR00243## 0.6 0.075 Ex. 31 89 ##STR00244## 3 0.9 Ex. 31 90
##STR00245## 0.4 0.09 Ex. 31 91 ##STR00246## 5 2 Ex. 31 92
##STR00247## >20 17 Ex. 31 93 ##STR00248## 0.35 0.05 Ex. 31 94
##STR00249## 0.4 0.03 Ex. 31 95 ##STR00250## 0.8 0.2 Ex. 31 96
##STR00251## 33% @ 5 .mu.M Ex. 31 97 ##STR00252## 0.8 0.28 Ex. 31
98 ##STR00253## 0.55 0.06 Ex. 31 99 ##STR00254## 0.9 0.05 Ex. 31
100 ##STR00255## 0.8 0.75 Ex. 31 101 ##STR00256## 0.3 0.04 Ex. 31
102 ##STR00257## 5.5 0.8 Ex. 31 103 ##STR00258## 0.7 0.05 Ex. 31
104 ##STR00259## 21% @ 5 .mu.M Ex. 31 105 ##STR00260## 0.55 0.2 Ex.
31 106 ##STR00261## 0.8 0.3 Ex. 31 107 ##STR00262## 0% @ 1 .mu.M 5
Ex. 31 108 ##STR00263## 10% @ 1 .mu.M 0.3 Ex. 31 109 ##STR00264##
32% @ 1 .mu.M 0.12 Ex. 31 110 ##STR00265## 0.7 0.55 Ex. 31 111
##STR00266## 0.4 0.095 Ex. 31 112 ##STR00267## 1.2 0.6 Ex. 31 113
##STR00268## 46% @ 1 .mu.M 0.2 Ex. 31 114 ##STR00269## 40% @ 1
.mu.M 0.1 Ex. 31 115 ##STR00270## 53% @ 1 .mu.M 0.1 Ex. 31 116
##STR00271## 4 Ex. 31 117 ##STR00272## 0% @ 20 .mu.M 1.9 Ex. 31 118
##STR00273## 0% @ 20 .mu.M 2.3 Ex. 31 119 ##STR00274## 3 Ex. 31 120
##STR00275## 0.12 0.01 Ex. 31 121 ##STR00276## 23 Ex. 31 122
##STR00277## 2.3 Ex. 31 123 ##STR00278## 1 Ex. 32 128 ##STR00279##
0.3 Ex. 32 129 ##STR00280## 3.0 Ex. 33 136 ##STR00281## 9 0.5 Ex.
34 139 ##STR00282## 44% @ 20 .mu.M Ex. 34 143 ##STR00283## 55% @ 20
.mu.M 2.4 Ex. 34 144 ##STR00284## 6% @ 20 .mu.M 6.9 Ex. 35 145
##STR00285## 3.8 0.84 Ex. 35 146 ##STR00286## 2.9 0.91 Ex. 35 147
##STR00287## 1.9 0.48 Ex. 36 148 ##STR00288## 5 2.0 Ex. 36 149
##STR00289## 8% @ 20 .mu.M 0.1 Ex. 36 150 ##STR00290## 10 1.0 Ex.
36 151 ##STR00291## 7.5 2.3 Ex. 36 152 ##STR00292## 35% @ 20 .mu.M
Ex. 36 153 ##STR00293## 5 4.8 Ex. 36 154 ##STR00294## .quadrature.
0.9 Ex. 36 155 ##STR00295## 39% @ 20 .mu.M Ex. 36 156 ##STR00296##
5 0.75 Ex. 36 157 ##STR00297## 6 2.4 Ex. 36 158 ##STR00298## >20
Ex. 36 159 ##STR00299## 1.5 Ex. 36 160 ##STR00300## 1.2 Ex. 36 161
##STR00301## 0.05 Ex. 36 162 ##STR00302## 0.04 Ex. 37 164
##STR00303## 5.0 Ex. 37 165 ##STR00304## 2.0 Ex. 37 166
##STR00305## Ex. 37 167 ##STR00306## Ex. 38 168 ##STR00307## 0% @
20 .mu.M 3 Ex. 39 170 ##STR00308## 48% @ 2 .mu.M 0.57 171
##STR00309## 20 172 ##STR00310## 10 173 ##STR00311## 35% @ 20 .mu.M
174 ##STR00312## >20 175 ##STR00313## >2 176 ##STR00314## 20%
@ 20 .mu.M 177 ##STR00315## 10% @ 20 .mu.M 178 ##STR00316## 2% @ 20
.mu.M >20
TABLE-US-00006 TABLE 4a rHDAC-1 rHDAC-8 Cpd. Structure IC.sub.50
(.mu.M) IC.sub.50 (.mu.M) 182 ##STR00317## 0.7 PI (64%) 1.3 PI
(74%) 179 ##STR00318## 0.5 0.53 183 ##STR00319## 0.38 0.41 184
##STR00320## 0.93 0.4 PI (79%) 185 ##STR00321## 0.84 0.61 187
##STR00322## 0.2 PI (62%) 0.21 PI = partial inhibition (100%
inhibition not reached - PI value indicates IC.sub.50 (at maximum %
inhibition reached))
Example 49
Inhibition of Histone Deacetylase in Whole Cells
1. Histone H4 Acetylation in Whole Cells by Immunoblots
[0904] T24 human bladder cancer cells growing in culture are
incubated with HDAC inhibitors for 16 hours. Histones are extracted
from the cells after the culture period as described by M. Yoshida
et al. (J. Biol. Chem. 265(28): 17174-17179 (1990)). 20 .mu..mu.g
of total histone protein are loaded onto SDS/PAGE and transferred
to nitrocellulose membranes. Membranes are probed with polyclonal
antibodies specific for acetylated histone H-4 (Upstate Biotech
Inc.), followed by horse radish peroxidase conjugated secondary
antibodies (Sigma). Enhanced Chemiluminescence (ECL) (Amersham)
detection is performed using Kodak films (Eastman Kodak).
Acetylated H-4 signal is quantified by densitometry.
[0905] Data for selected compounds are presented in Table 5. Data
are presented as the concentration effective for reducing the
acetylated H-4 signal by 50% (EC.sub.50).
TABLE-US-00007 TABLE 5 Inhibibition of Histone Acetylation in Cells
Cpd. Structure EC.sub.50 (.mu.M) 36 ##STR00323## 5 90 ##STR00324##
1 98 ##STR00325## 1 107 ##STR00326## 5 118 ##STR00327## 3 120
##STR00328## 1 122 ##STR00329## 2
2. Acid Urea Triton (AUT) Gel Analysis of Histone Acetylation.
[0906] Human cancer cells (T24, 293T or Jurkat cells) growing in
culture are incubated with HDAC inhibitors for 24 h. Histones are
extracted from the cells as described by M. Yoshida et al. (J.
Biol. Chem. 265(28): 17174-17179 (1990)). Acid urea triton (AUT)
gel electrophoresis is used for detection of acetylated histone
molecules. Histones (150 .mu..mu.g of total protein) are
electrophoresed at 80 V for 16 h at room temperature as described
by M. Yoshida et al., supra. Gels are stained with Coomassie
brilliant blue to visualize histones, dried and scanned by
densitometry to quantified acetylation of histones.
Example 50
Antineoplastic Effect of Histone Deacetylase Inhibitors on Tumor
Cells In Vivo
[0907] Eight to ten week old female BALB/c nude mice (Taconic Labs,
Great Barrington, N.Y.) are injected subcutaneously in the flank
area with 2.times.10.sup.6 preconditioned A549 human lung carcinoma
cells. Preconditioning of these cells is done by a minimum of three
consecutive tumor transplantations in the same strain of nude mice.
Subsequently, tumor fragments of approximately 30 mgs are excised
and implanted subcutaneously in mice, in the left flank area, under
Forene anesthesia (Abbott Labs, Geneve, Switzerland). When the
tumors reach a mean volume of 100 mm.sup.3, the mice are treated
intravenously, subcutaneously, or intraperitoneally by daily
injection, with a solution of the histone deacetylase inhibitor in
an appropriate vehicle, such as PBS, DMSO/water, or Tween 80/water,
at a starting dose of 10 mg/kg. The optimal dose of the HDAC
inhibitor is established by dose response experiments according to
standard protocols. Tumor volume is calculated every second day
post infusion according to standard methods (e.g., Meyer et al.,
Int. J. Cancer 43: 851-856 (1989)). Treatment with the HDAC
inhibitors according to the invention causes a significant
reduction in tumor weight and volume relative to controls treated
with saline only (i.e., no HDAC inhibitor). In addition, the
activity of histone deacetylase when measured is expected to be
significantly reduced relative to saline treated controls.
Example 51
Synergistic Antineoplastic Effect of Histone Deacetylase Inhibitors
and Histone Deacetylase Antisense Oligonucleotides on Tumor Cells
In Vivo
[0908] The purpose of this example is to illustrate the ability of
the histone deacetylase inhibitor of the invention and a histone
deacetylase antisense oligonucleotide to synergistically inhibit
tumor growth in a mammal. Preferably, the antisense oligonucleotide
and the HDAC inhibitor inhibit the expression and activity of the
same histone deacetylase.
[0909] As described in Example 10, mice bearing implanted A549
tumors (mean volume 100 mm.sup.3) are treated daily with saline
preparations containing from about 0.1 mg to about 30 mg per kg
body weight of histone deacetylase antisense oligonucleotide. A
second group of mice is treated daily with pharmaceutically
acceptable preparations containing from about 0.01 mg to about 5 mg
per kg body weight of HDAC inhibitor.
[0910] Some mice receive both the antisense oligonucleotide and the
HDAC inhibitor. Of these mice, one group may receive the antisense
oligonucleotide and the HDAC inhibitor simultaneously intravenously
via the tail vein. Another group may receive the antisense
oligonucleotide via the tail vein, and the HDAC inhibitor
subcutaneously. Yet another group may receive both the antisense
oligonucleotide and the HDAC inhibitor subcutaneously. Control
groups of mice are similarly established which receive no treatment
(e.g., saline only), a mismatch antisense oligonucleotide only, a
control compound that does not inhibit histone deacetylase
activity, and a mismatch antisense oligonucleotide with a control
compound.
[0911] Tumor volume is measured with calipers. Treatment with the
antisense oligonucleotide plus the histone deacetylase protein
inhibitor according to the invention causes a significant reduction
in tumor weight and volume relative to controls.
Example 52
Solubility
[0912] The solubility of several compounds according to the
invention was measured, and the results are displayed in the table
below.
TABLE-US-00008 ##STR00330## Solubility (.mu.M) Q pH 2 Water* H 3 3
##STR00331## 4 5 ##STR00332## 132 195 ##STR00333## 10 9
##STR00334## >250 >250 ##STR00335## 142 136 ##STR00336## 186
130 ##STR00337## 49 ##STR00338## >2000 >1000 ##STR00339## 166
165 *Milli-Q .RTM. purified water
[0913] While the invention has been described in connection with
specific embodiments thereof, it will be understood that it is
capable of further modifications and this application is intended
to cover any variations, uses, or adaptations of the invention
following, in general, the principles of the invention and
including such departures from the present disclosure as come
within known or customary practice within the art to which the
invention pertains and as may be applied to the essential features
hereinbefore set forth, and as follows in the scope of the appended
claims.
Sequence CWU 1
1
21120DNAArtificialSynthetic; antisense oligonucleotide targeted to
human HDAC-1 1gagacagcag caccagcggg 20220DNAArtificialSynthetic;
antisense oligonucleotide targeted to human HDAC-1 2atgaccgagt
gggagacagc 20320DNAArtificialSynthetic; antisense oligonucleotide
targeted to human HDAC-1 3ggatgaccga gtgggagaca
20420DNAArtificialSynthetic; antisense oligonucleotide targeted to
human HDAC-1 4caggatgacc gagtgggaga 20520DNAArtificialSynthetic;
antisense oligonucleotide targeted to human HDAC-1 5tgtgttctca
ggatgaccga 20620DNAArtificialSynthetic; antisense oligonucleotide
targeted to human HDAC-1 6gagtgacaga gacgctcagg
20720DNAArtificialSynthetic; antisense oligonucleotide targeted to
human HDAC-1 7ttctggcttc tcctccttgg 20820DNAArtificialSynthetic;
antisense oligonucleotide targeted to human HDAC-1 8cttgacctcc
tccttgaccc 20920DNAArtificialSynthetic; antisense oligonucleotide
targeted to human HDAC-1 9ggaagccaga gctggagagg
201020DNAArtificialSynthetic; antisense oligonucleotide targeted to
human HDAC-1 10gaaacgtgag ggactcagca 201123DNAArtificialSynthetic;
antisense oligonucleotide targeted to human HDAC-1 11ccgtcgtagt
agtaacagac ttt 231222DNAArtificialSynthetic; antisense
oligonucleotide targeted to human HDAC-1 12tgtccataat agtaatttcc aa
221326DNAArtificialSynthetic; antisense oligonucleotide targeted to
human HDAC-1 13cagcaaatta tgagtcatgc ggattc
261420DNAArtificialSynthetic; antisense oligonucleotide targeted to
human HDAC-2 14ctccttgact gtacgccatg 201520DNAArtificialSynthetic;
antisense oligonucleotide targeted to human HDAC-2 15tgctgctgct
gctgctgccg 201620DNAArtificialSynthetic; antisense oligonucleotide
targeted to human HDAC-2 16cctcctgctg ctgctgctgc
201723DNAArtificialSynthetic; antisense oligonucleotide targeted to
human HDAC-2 17ccgtcgtagt agtagcagac ttt
231822DNAArtificialSynthetic; antisense oligonucleotide targeted to
human HDAC-2 18tgtccataat aataatttcc aa
221926DNAArtificialSynthetic; antisense oligonucleotide targeted to
human HDAC-2 19cagcaagtta tgggtcatgc ggattc
262020DNAArtificialSynthetic; antisense oligonucleotide targeted to
human HDAC-2 20ggttcctttg gtatctgttt 202120DNAArtificialSynthetic;
antisense oligonucleotide targeted to human HDAC-4 21gctgcctgcc
gtgcccaccc 20
* * * * *