U.S. patent application number 15/355959 was filed with the patent office on 2017-03-09 for novel molecules that selectively inhibit histone deacetylase 6 relative to histone deacetylase.
This patent application is currently assigned to The Trustees of Columbia University in the City of New York. The applicant listed for this patent is Ronald Breslow, Paul A. Marks. Invention is credited to Ronald Breslow, Paul A. Marks.
Application Number | 20170066712 15/355959 |
Document ID | / |
Family ID | 48044052 |
Filed Date | 2017-03-09 |
United States Patent
Application |
20170066712 |
Kind Code |
A1 |
Breslow; Ronald ; et
al. |
March 9, 2017 |
NOVEL MOLECULES THAT SELECTIVELY INHIBIT HISTONE DEACETYLASE 6
RELATIVE TO HISTONE DEACETYLASE
Abstract
This invention provides a compound having the structure:
##STR00001## wherein R.sub.1 is H, halogen, --NR.sub.5R.sub.6,
--NR.sub.5--C(.dbd.0)-R.sub.6, --NH--C(.dbd.O)--OR.sub.7,
--OR.sub.7, --NO.sub.2, --CN, --SR.sub.7, --SO.sub.2R.sub.7,
--CO.sub.2R.sub.7, CF.sub.3, --SOR.sub.7, --POR.sub.7,
--C(.dbd.S)R.sub.7, --C(.dbd.O)--NR.sub.5R.sub.6,
--CH.sub.2--C(.dbd.O)--NR.sub.5R.sub.6, --C(.dbd.NR.sub.5)R.sub.6,
--P(.dbd.O)(OR.sub.5)(OR.sub.6), --P(OR.sub.5)(OR.sub.6),
--C(.dbd.S)R.sub.7, C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5
alkynyl, aryl, heteroaryl, or heterocyclyl, wherein R.sub.5,
R.sub.6, and R.sub.7 and are each, independently, H, C.sub.1-5
alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, heteroalkyl,
cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; m is an integer
from 0 to 2; R.sub.2 and R.sub.3 are each, independently, H,
halogen, --NH.sub.2, --CX.sub.3, --C(.dbd.O)OR.sub.8,
C(.dbd.O)R.sub.8, --C(.dbd.O)NR.sub.9R.sub.10, C.sub.1-10 alkyl,
C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, heteroalkyl, aryl,
heteroaryl, or heterocyclyl; wherein X is Cl, Br, or F; R.sub.8,
R.sub.9 and R.sub.10 are each, independently, H, C.sub.1-5 alkyl,
C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, heteroalkyl, cycloalkyl,
heterocycloalkyl, aryl, or heteroaryl; Q is --Ar.sub.1--Z-- or
--Z--Ar.sub.1--Z--, wherein Ar.sub.1 is aryl or heteroaryl; and
each occurrence of Z is independently present or absent, and when
present is --O--, --S--, --CH.sub.2--, --C(O)--, --NH--,
--NH--NH--, --NHC(.dbd.O)--, --C(.dbd.O)NH--,
--NHC(.dbd.O)CH.sub.2NH--, --NHC(.dbd.O)CH.sub.2C(.dbd.O)--,
--N(OH)--, --CH.sub.2CH.sub.2-- or --NHC(.dbd.O)CH.dbd.CH--; and
R.sub.4 is alkyl, --OR.sub.11 or --NH--OR.sub.11, wherein R.sub.11
is H, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
aryl, heteroaryl, or heterocyclyl, and when Q is --Ar.sub.1--Z--, Z
is absent, Ar.sub.1 is phenyl, R.sub.2 and R.sub.3 are H, n=1, and
R.sub.4 is --NHOH, then R.sub.1 is other than carbazole,
tetrahydro-.beta.-carboline, tetrahydro-.gamma.-carboline,
--C(.dbd.O)--NR.sub.5R.sub.6 and --NR.sub.5--C(.dbd.0)-R.sub.6,
wherein one of R.sub.5 or R.sub.6 is quinline and the other of
R.sub.5 or R.sub.6 is H; or a pharmaceutically acceptable salt
thereof.
Inventors: |
Breslow; Ronald; (New York,
NY) ; Marks; Paul A.; (Washington, CT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Breslow; Ronald
Marks; Paul A. |
New York
Washington |
NY
CT |
US
US |
|
|
Assignee: |
The Trustees of Columbia University
in the City of New York
New York
NY
Sloan-Kettering Institute for Cancer Research
New York
NY
|
Family ID: |
48044052 |
Appl. No.: |
15/355959 |
Filed: |
November 18, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14349291 |
Apr 2, 2014 |
9499479 |
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PCT/US12/00459 |
Oct 3, 2012 |
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15355959 |
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61620783 |
Apr 5, 2012 |
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61542598 |
Oct 3, 2011 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07C 259/10 20130101;
C07D 209/18 20130101; C07C 259/06 20130101; C07C 2602/10 20170501;
C07D 209/42 20130101; C07D 209/24 20130101; C07D 215/40 20130101;
C07D 235/06 20130101; A61P 25/28 20180101; C07D 215/06 20130101;
C07D 209/08 20130101; C07C 275/64 20130101 |
International
Class: |
C07C 259/10 20060101
C07C259/10; C07C 275/64 20060101 C07C275/64; C07D 235/06 20060101
C07D235/06; C07D 209/42 20060101 C07D209/42; C07D 209/24 20060101
C07D209/24; C07D 215/40 20060101 C07D215/40; C07C 259/06 20060101
C07C259/06 |
Claims
1-35. (canceled)
36. A compound having the structure: ##STR00058## wherein R.sub.1
is --NR.sub.5--C(.dbd.0)-R.sub.6 or --C(.dbd.O)--NR.sub.5R.sub.6,
wherein R.sub.5 is ##STR00059## R.sub.6 is ##STR00060## wherein X
is a Cl, Br, or F; and R.sub.12 is H, C.sub.1-5 alkyl, C.sub.2-5
alkenyl, C.sub.2-5 alkynyl, heteroalkyl, cycloalkyl,
heterocycloalkyl, aryl, or heteroaryl; m is an integer from 0 to 2;
R.sub.2 and R.sub.3 are each, independently, H, halogen,
--NH.sub.2, --CX.sub.3, --C(.dbd.O)OR.sub.8, C(.dbd.O)R.sub.8,
--C(.dbd.O) NR.sub.9R.sub.10, C.sub.1-10 alkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, heteroalkyl, aryl, heteroaryl, or heterocyclyl;
wherein X is Cl, Br, or F; R.sub.8, R.sub.9 and R.sub.10 are each,
independently, H, C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5
alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or
heteroaryl; Q is --Ar.sub.1--Z-- or --Z--Ar.sub.1--Z--, wherein
Ar.sub.1 is ##STR00061## wherein X is a Cl, Br, or F; and each
occurrence of Z is present or absent, and when present is --O--,
--S--, --CH.sub.2--, --C(O)--, --NH--, --NH--NH--, --NHC(.dbd.O)--,
--C(.dbd.O)NH--, --NHC(.dbd.O)CH.sub.2NH--,
--NHC(.dbd.O)CH.sub.2C(.dbd.O)--, --N(OH)--, --CH.sub.2CH.sub.2--
or --NHC(.dbd.O)CH.dbd.CH--; and R.sub.4 is --NH--OR.sub.11,
wherein R.sub.11 is H, C.sub.1-10 alkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, aryl, heteroaryl, or heterocyclyl, or a
pharmaceutically acceptable salt thereof.
37. The compound of claim 36 having the structure: ##STR00062##
wherein R.sub.1 is --NR.sub.5--C(.dbd.0)-R.sub.6 or
--C(.dbd.O)--NR.sub.5R.sub.6, wherein R.sub.5 is ##STR00063##
R.sub.6 is ##STR00064## wherein X is a Cl, Br, or F; and R.sub.12
is H, C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl,
heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; m
is an integer from 0 to 2; R.sub.2 and R.sub.3 are each,
independently, H, halogen, --NH.sub.2, --CX.sub.3,
--C(.dbd.O)OR.sub.8, C(.dbd.O)R.sub.8, --C(.dbd.O)NR.sub.9R.sub.10,
C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
heteroalkyl, aryl, heteroaryl, or heterocyclyl; wherein X is Cl,
Br, or F; R.sub.8, R.sub.9 and R.sub.10 are each, independently, H,
C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, heteroalkyl,
cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; Q is
--Ar.sub.1--Z-- or --Z--Ar.sub.1--Z--, wherein Ar.sub.1 is
##STR00065## wherein X is a Cl, Br, or F; and each occurrence of Z
is present or absent, and when present is --O--, --S--,
--CH.sub.2--, --C(O)--, --NH--, --NH--NH--, --NHC(.dbd.O)--,
--C(.dbd.O)NH--, --NHC(.dbd.O)CH.sub.2NH--,
--NHC(.dbd.O)CH.sub.2C(.dbd.O)--, --N(OH)--, --CH.sub.2CH.sub.2--
or --NHC(.dbd.O)CH.dbd.CH--; and R.sub.4 is --NH--OR.sub.11,
wherein R.sub.11 is H, C.sub.1-10 alkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, aryl, heteroaryl, or heterocyclyl, and wherein
at least one Z is present, or a pharmaceutically acceptable salt
thereof.
38. The compound of claim 37 having the structure: ##STR00066##
wherein R.sub.1 is --NR.sub.5--C(.dbd.0)-R.sub.6 or
--C(.dbd.O)--NR.sub.5R.sub.6, wherein R.sub.5 is ##STR00067##
R.sub.6 is ##STR00068## wherein X is a Cl, Br, or F; and R.sub.12
is H, C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl,
heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; m
is an integer from 0 to 2; R.sub.2 and R.sub.3 are each,
independently, H, halogen, --NH.sub.2, --CX.sub.3,
--C(.dbd.O)OR.sub.8, C(.dbd.O)R.sub.8, --C(.dbd.O)NR.sub.9R.sub.10,
C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
heteroalkyl, aryl, heteroaryl, or heterocyclyl; wherein X is Cl,
Br, or F; R.sub.8, R.sub.9 and R.sub.10 are each, independently, H,
C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, heteroalkyl,
cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; Ar.sub.1 is
##STR00069## wherein X is a Cl, Br, or F; Z is --O--, --S--,
--CH.sub.2--, --C(O)--, --NH--, --NH--NH--, --NHC(.dbd.O)--,
--C(.dbd.O)NH--, --NHC(.dbd.O)CH.sub.2NH-- or
--NHC(.dbd.O)CH.dbd.CH--; and R.sub.4 is --NH--OR.sub.11, wherein
R.sub.11 is H, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10
alkynyl, aryl, heteroaryl, or heterocyclyl, or a pharmaceutically
acceptable salt thereof.
39. The compound of claim 37, wherein m=1; R.sub.2 is H or
CH.sub.3; and R.sub.3 is H, CH.sub.3, Cl, Br, F, or CF.sub.3, or a
pharmaceutically acceptable salt thereof.
40. The compound of claim 37, wherein R.sub.4 is --NH--OR.sub.11,
wherein R.sub.11 is H or CH.sub.3, or a pharmaceutically acceptable
salt thereof.
41. The compound of claim 37, wherein R.sub.5 is ##STR00070## and
R.sub.6 is ##STR00071## or a pharmaceutically acceptable salt
thereof.
42. The compound of claim 37 having the structure: ##STR00072##
##STR00073## ##STR00074## ##STR00075## ##STR00076## or a
pharmaceutically acceptable salt thereof.
43. A pharmaceutical composition comprising the compound of claim
37 and a pharmaceutically acceptable carrier.
44. A method of inhibiting the activity of a histone deactylase in
a cell, the method comprising contacting the histone deacetylase
with the compound of claim 37 so as to inhibit the activity of the
histone deacetylase; or of increasing accumulation of acetylated
alpha tubulin in a cell, the method comprising contacting the cell
with the compound of claim 37 so as to increase the accumulation of
acetylated alpha-tubulin in the cell.
45. The compound of claim 36 having the structure: ##STR00077##
wherein R.sub.1 is --NR.sub.5--C(.dbd.0)-R.sub.6 or
--C(.dbd.O)--NR.sub.5R.sub.6, wherein R.sub.5 is ##STR00078##
R.sub.6 is ##STR00079## wherein X is a Cl, Br, or F; and R.sub.12
is H, C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl,
heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; m
is an integer from 0 to 2; R.sub.2 and R.sub.3 are each,
independently, H, halogen, --NH.sub.2, --CX.sub.3,
--C(.dbd.O)OR.sub.8, C(.dbd.O)R.sub.8, --C(.dbd.O)NR.sub.9R.sub.10,
C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
heteroalkyl, aryl, heteroaryl, or heterocyclyl; wherein X is Cl,
Br, or F; R.sub.8, R.sub.9 and R.sub.10 are each, independently, H,
C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, heteroalkyl,
cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; Q is
--Ar.sub.1--Z-- or --Z--Ar.sub.1--Z--, wherein Ar.sub.1 is
##STR00080## wherein X is a Cl, Br, or F; and each occurrence of Z
absent; and R.sub.4 is --NH--OR.sub.11, wherein R.sub.11 is H,
C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, aryl,
heteroaryl, or heterocyclyl, or a pharmaceutically acceptable salt
thereof.
46. The compound of claim 45 having the structure: ##STR00081##
wherein R.sub.1 is --NR.sub.5--C(.dbd.0)-R.sub.6 or
--C(.dbd.O)--NR.sub.5R.sub.6, wherein R.sub.5 is ##STR00082##
R.sub.6 is ##STR00083## wherein X is a Cl, Br, or F; and R.sub.12
is H, C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl,
heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; m
is an integer from 0 to 2; R.sub.2 and R.sub.3 are each,
independently, H, halogen, --NH.sub.2, --CX.sub.3,
--C(.dbd.O)OR.sub.8, C(.dbd.O)R.sub.8, --C(.dbd.O)NR.sub.9R.sub.10,
C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
heteroalkyl, aryl, heteroaryl, or heterocyclyl; wherein X is Cl,
Br, or F; R.sub.8, R.sub.9 and R.sub.10 are each, independently, H,
C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, heteroalkyl,
cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; Ar.sub.1 is
##STR00084## wherein X is a Cl, Br, or F; Z absent; and R.sub.4 is
--NH--OR.sub.11, wherein R.sub.11 is H, C.sub.1-10 alkyl,
C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, aryl, heteroaryl, or
heterocyclyl, or a pharmaceutically acceptable salt thereof.
47. The compound of claim 45, wherein m=1; R.sub.2 is H or
CH.sub.3; and R.sub.3 is H, CH.sub.3, Cl, Br, F, or CF.sub.3, or a
pharmaceutically acceptable salt thereof.
48. The compound of claim 45, wherein R.sub.4 is --NH--OR.sub.11,
wherein R.sub.11 is H or CH.sub.3, or a pharmaceutically acceptable
salt thereof.
49. The compound of claim 45, wherein R.sub.5 is ##STR00085## and
R.sub.6 is ##STR00086## or a pharmaceutically acceptable salt
thereof.
50. A pharmaceutical composition comprising the compound of claim
45 and a pharmaceutically acceptable carrier.
51. A method of inhibiting the activity of a histone deactylase in
a cell, the method comprising contacting the histone deacetylase
with the compound of claim 45 so as to inhibit the activity of the
histone deacetylase; or of increasing accumulation of acetylated
alpha tubulin in a cell, the method comprising contacting the cell
with the compound of claim 45 so as to increase the accumulation of
acetylated alpha-tubulin in the cell.
52. A compound having the structure: ##STR00087## ##STR00088## or a
pharmaceutically acceptable salt thereof.
53. A pharmaceutical composition comprising the compound of claim
52 and a pharmaceutically acceptable carrier.
54. A method of inhibiting the activity of a histone deactylase in
a cell, the method comprising contacting the histone deacetylase
with the compound of claim 52 so as to inhibit the activity of the
histone deacetylase; or of increasing accumulation of acetylated
alpha tubulin in a cell, the method comprising contacting the cell
with the compound of claim 52 so as to increase the accumulation of
acetylated alpha-tubulin in the cell.
Description
[0001] This application claims priority of U.S. Provisional
Application No. 61/620,783, filed Apr. 5, 2012 and 61/542,598,
filed Oct. 3, 2011, the contents of each of which are hereby
incorporated by reference.
[0002] Throughout this application, certain publications are
referenced in parentheses. Full citations for these publications
may be found immediately preceding the claims. The disclosures of
these publications in their entireties are hereby incorporated by
reference into this application in order to describe more fully the
state of the art to which this invention relates.
BACKGROUND OF THE INVENTION
[0003] To date, eighteen histone deacetylases (HDACs) have been
identified in humans. Eleven HDACs (HDAC1-11) are zinc-dependent
and seven HDACs, designated sirtuins 1-7, are NAD+-dependent (1).
Aberrant activity of HDACs has been implicated in many disease
states, including cancer (2). When zinc-dependent HDACs are
inhibited, the levels of acetylation of certain proteins are
elevated, with many resulting physiological effects. Many
inhibitors of HDACs have been developed for use against cancers and
other disease states. One well-known HDAC inhibitor,
suberoylanilide hydroxamic acid (SAHA, Vorinostat), was approved in
2006 for human use following the results of more than 100 human
trials against various forms of cancer and is currently in use.
Phase I, II and III clinical trials with vorinostat as single
therapy and in combination therapy with various anti-cancer agents
for hematologic and solid neoplasms are ongoing.
[0004] While HDACs are associated with deacetylation of histones in
the context of gene expression and chromatin remodeling, there is
abundant evidence indicating that not all functions of HDACs are
dedicated to deacetylation of histones. Rather, some HDACs have
been shown to exert deacetylase activity on proteins other than
histones. One such HDAC is HDAC6, a cytoplasmic,
microtubule-associated deacetylase, which has been found to
regulate microtubule acetylation and chemotactic cell motility
(3).
[0005] HDAC6 is predominantly a cytoplasmic, microtubule-associated
member of the class IIB family of histone deacetylases. HDAC6
possesses two catalytic domains, a ubiquitin-binding domain and a
C-terminal zinc finger domain (4). HDAC6 catalyzes deacetylation of
cytoplasmic protein substrates, such as .alpha.-tubulin, Hsp90,
peroxiredoxins, and cortactin (4). HDAC6 has also been demonstrated
to direct misfolded protein aggregates into aggresomes, which are
major repositories formed to manage excessive levels of misfolded
and aggregated protein for eventual elimination. Aggresomes are of
clinical interest as they are similar to cytoplasmic inclusion
bodies commonly observed in neurodegenerative diseases (5).
[0006] Haggarty et al. (6) have shown that the C-terminal catalytic
domain of HDAC6, the domain responsible for .alpha.-tubulin
deacetylation, can be inhibited by the small-molecule inhibitor,
tubacin. Haggarty et al found that the inhibition of HDAC6 with
tubacin did not affect the stability of microtubules, but decreased
cell motility. Given the dependence of metastasis and angiogenesis
on cell movement, increasing the acetylation level of
.alpha.-tubulin may be an important component to the antimetastatic
and antiangiogenic activities of HDAC inhibitors (6).
[0007] Heat shock protein 90 (Hsp90) is an important chaperone
protein involved in protein folding and is overexpressed in many
cancer cell types (2, 7). The disruption of the folding and
chaperoning functions of Hsp90 causes its client proteins to be
destabilized and eventually degraded. HDAC6 is an attractive target
for cancer treatment because acetylated Hsp90 has a reduced ability
to perform its chaperoning function (2, 7), with consequent
activation of the intrinsic pathway of apoptosis.
[0008] In general, for diseases caused by aberrant gene
transcription, the most effective treatment would involve targeting
only the genes relevant to the disease (2). In the context of HDAC
inhibitor treatment, this would involve inhibiting only those HDAC
isoforms relevant to the disease state, thereby minimizing changes
not related to the disease, and possibly reducing side effects and
toxicity. While SAHA combines efficacy with minimum toxicity, its
inhibitory activity is not selective among the known human
HDACs.
[0009] Marks & Breslow (8, 9) describes the development of HDAC
inhibitor voronistat as an anti-cancer drug. HDAC inhibitors have
also been identified as a correction for cholesterol and
sphingolipid transport defects in human Niemann-Pick type C disease
(10).
[0010] In view of the importance of inhibiting only those HDAC
isoforms relevant to a disease state, minimizing acetylation of
proteins not related to the disease, and reducing side effects and
toxicity, new HDAC inhibitors that are selective for specific HDACs
are needed. Herein, new selective HDAC inhibitors are
described.
SUMMARY OF THE INVENTION
[0011] This invention provides a compound having the structure:
##STR00002## [0012] wherein [0013] R.sub.1 is H, halogen,
--NR.sub.5R.sub.6, --NR.sub.5--C(.dbd.0)-R.sub.6,
--NH--C(.dbd.O)--OR.sub.7, --OR.sub.7, --NO.sub.2, --CN,
--SR.sub.7, --SO.sub.2R.sub.7, --CO.sub.2R.sub.7, CF.sub.3,
--SOR.sub.7, --POR.sub.7, --C(.dbd.S)R.sub.7,
--C(.dbd.O)--NR.sub.5R.sub.6,
--CH.sub.2--C(.dbd.O)--NR.sub.5R.sub.6, --C(.dbd.NR.sub.5)R.sub.6,
--P(.dbd.O)(OR.sub.5)(OR.sub.6), --P(OR.sub.5)(OR.sub.6),
--C(.dbd.S)R.sub.7, C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5
alkynyl, aryl, heteroaryl, or heterocyclyl, [0014] wherein R.sub.5,
R.sub.6, and R.sub.7 and are each, independently, H, C.sub.1-5
alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, heteroalkyl,
cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; [0015] m is an
integer from 0 to 2; [0016] R.sub.2 and R.sub.3 are each,
independently, H, halogen, --NH.sub.2, --CX.sub.3,
--C(.dbd.O)OR.sub.8, C(.dbd.O)R.sub.8, --C(.dbd.O)NR.sub.9R.sub.10,
C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
heteroalkyl, aryl, heteroaryl, or heterocyclyl; [0017] wherein
[0018] X is Cl, Br, or F; [0019] R.sub.8, R.sub.9 and R.sub.10 are
each, independently, H, C.sub.1-5 alkyl, C.sub.2-5 alkenyl,
C.sub.2-5 alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,
or heteroaryl; [0020] Q is --Ar.sub.1--Z-- or --Z--Ar.sub.1--Z--,
[0021] wherein Ar.sub.1 is aryl or heteroaryl; and [0022] each
occurrence of Z is independently present or absent, and when
present is --O--, --S--, --CH.sub.2--, --C(O)--, --NH--,
--NH--NH--, --NHC(.dbd.O)--, --C(.dbd.O)NH--,
--NHC(.dbd.O)CH.sub.2NH--, --NHC(.dbd.O)CH.sub.2C(.dbd.O)--,
--N(OH)--, --CH.sub.2CH.sub.2-- or --NHC(.dbd.O)CH.dbd.CH--; and
[0023] R.sub.4 is alkyl, --OR.sub.11 or --NH--OR.sub.11, [0024]
wherein R.sub.11 is H, C.sub.1-10 alkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, aryl, heteroaryl, or heterocyclyl, and [0025]
when Q is --Ar.sub.1--Z--, Z is absent, Ar.sub.1 is phenyl, R.sub.2
and R.sub.3 are H, n=1, and R.sub.4 is --NHOH, then R.sub.1 is
other than carbazole, tetrahydro-.beta.-carboline,
tetrahydro-.gamma.-carboline, --C(.dbd.O)--NR.sub.5R.sub.6 and
--NR.sub.5--C(.dbd.0)-R.sub.6, wherein one of R.sub.5 or R.sub.6 is
quinoline and the other of R.sub.5 or R.sub.6 is H; or a
pharmaceutically acceptable salt thereof.
BRIEF DESCRIPTION OF THE FIGURES
[0026] FIG. 1. Schematic representation of HDAC6.
[0027] FIG. 2. Blot showing accumulation of acetylated
alpha-tubulin and acetylated histone H3 in LNCaP cells cultured
with compound 6. GADPH used as loading control.
[0028] FIG. 3. Cell growth assay data in LNCaP cells with compound
6.
[0029] FIG. 4. Cell viability assay data in LNCaP cells with
compound 6.
DETAILED DESCRIPTION OF THE INVENTION
[0030] This invention provides a compound having the structure:
##STR00003## [0031] wherein [0032] R.sub.1 is H, halogen,
--NR.sub.5R.sub.6, --NR.sub.5--C(.dbd.0)-R.sub.6,
--NH--C(.dbd.O)--OR.sub.7, --OR.sub.7, --NO.sub.2, --CN,
--SR.sub.7, --SO.sub.2R.sub.7, --CO.sub.2R.sub.7, CF.sub.3,
--SOR.sub.7, --POR.sub.7, --C(.dbd.S)R.sub.7,
--C(.dbd.O)--NR.sub.5R.sub.6,
--CH.sub.2--C(.dbd.O)--NR.sub.5R.sub.6, --C(.dbd.NR.sub.5)R.sub.6,
--P(.dbd.O)(OR.sub.5)(OR.sub.6), --P(OR.sub.5)(OR.sub.6),
--C(.dbd.S)R.sub.7, C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5
alkynyl, aryl, heteroaryl, or heterocyclyl, [0033] wherein R.sub.5,
R.sub.6, and R.sub.7 and are each, independently, H, C.sub.1-5
alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, heteroalkyl,
cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; [0034] m is an
integer from 0 to 2; [0035] R.sub.2 and R.sub.3 are each,
independently, H, halogen, --NH.sub.2, --CX.sub.3,
--C(.dbd.O)OR.sub.8, C(.dbd.O)R.sub.8, --C(.dbd.O)NR.sub.9R.sub.10,
C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
heteroalkyl, aryl, heteroaryl, or heterocyclyl; [0036] wherein
[0037] X is Cl, Br, or F; [0038] R.sub.8, R.sub.9 and R.sub.10 are
each, independently, H, C.sub.1-5 alkyl, C.sub.2-5 alkenyl,
C.sub.2-5 alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,
or heteroaryl; [0039] Q is --Ar.sub.1--Z-- or --Z--Ar.sub.1--Z--,
[0040] wherein Ar.sub.1 is aryl or heteroaryl; and [0041] each
occurrence of Z is independently present or absent, and when
present is --O--, --S--, --CH.sub.2--, --C(O)--, --NH--,
--NH--NH--, --NHC(.dbd.O)--, --C(.dbd.O)NH--,
--NHC(.dbd.O)CH.sub.2NH--, --NHC(.dbd.O)CH.sub.2C(.dbd.O)--,
--N(OH)--, --CH.sub.2CH.sub.2-- or --NHC(.dbd.O)CH.dbd.CH--; and
[0042] R.sub.4 is alkyl, --OR.sub.11 or --NH--OR.sub.11, [0043]
wherein R.sub.11 is H, C.sub.1-10 alkyl, C.sub.2-10 alkenyl,
C.sub.2-10 alkynyl, aryl, heteroaryl, or heterocyclyl, and [0044]
when Q is --Ar.sub.1--Z--, Z is absent, Ar.sub.1 is phenyl, R.sub.2
and R.sub.3 are H, n=1, and R.sub.4 is --NHOH, then R.sub.1 is
other than carbazole, tetrahydro-.beta.-carboline,
tetrahydro-.gamma.-carboline, --C(.dbd.O)--NR.sub.5R.sub.6 and
--NR.sub.5--C(.dbd.0)-R.sub.6, wherein one of R.sub.5 or R.sub.6 is
quinoline and the other of R.sub.5 or R.sub.6 is H; or a
pharmaceutically acceptable salt thereof.
[0045] In some embodiments, the compound wherein [0046] wherein Q
is --Ar.sub.1--Z--, --Z--Ar.sub.1-- or --Z--Ar.sub.1--Z-- or a
pharmaceutically acceptable salt thereof.
[0047] This invention provides a compound having the structure:
##STR00004## [0048] wherein [0049] R.sub.1 is H, halogen,
--NR.sub.5R.sub.6, --NR.sub.5--C(.dbd.O)--R.sub.6,
--NH--C(.dbd.O)--OR.sub.7, --OR.sub.7, --NO.sub.2, --CN,
--SR.sub.7, --SO.sub.2R.sub.7, --CO.sub.2R.sub.7, CF.sub.3,
--SOR.sub.7, --POR.sub.7, --C(.dbd.S)R.sub.7,
--C(.dbd.O)--NR.sub.5R.sub.6,
--CH.sub.2--C(.dbd.O)--NR.sub.5R.sub.6, --C(.dbd.NR.sub.5)R.sub.6,
--P(.dbd.O)(OR.sub.5)(OR.sub.6), --P(OR.sub.5)(OR.sub.5)(OR.sub.6),
--C(.dbd.S)R.sub.7, C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5
alkynyl, aryl, heteroaryl, or heterocyclyl, [0050] wherein R.sub.5,
R.sub.6, and R.sub.7 and are each, independently, H, C.sub.1-5
alkyl, C.sub.2-5 alkenyl, C.sub.2-5 alkynyl, heteroalkyl,
cycloalkyl, heterocycloalkyl, aryl, or heteroaryl; [0051] m is an
integer from 0 to 2; [0052] R.sub.2 and R.sub.3 are each,
independently, H, halogen, --NH.sub.2, --CX.sub.3,
--C(.dbd.O)OR.sub.8, C(.dbd.O)R.sub.8, --C(.dbd.O)NR.sub.9R.sub.10,
C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
heteroalkyl, aryl, heteroaryl, or heterocyclyl; [0053] wherein
[0054] X is Cl, Br, or F; [0055] R.sub.8, R.sub.9 and R.sub.10 are
each, independently, H, C.sub.1-5 alkyl, C.sub.2-5 alkenyl,
C.sub.2-5 alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,
or heteroaryl; [0056] Ar.sub.1 is aryl or heteroaryl; [0057] Z is
present or absent, and when present is --O--, --S--, --CH.sub.2--,
--C(O)--, --NH--, --NH--NH--, --NHC(.dbd.O)--, --C(.dbd.O)NH--,
--NHC(.dbd.O)CH.sub.2NH-- or --NHC(.dbd.O)CH.dbd.CH--; and [0058]
R.sub.4 is --OR.sub.11 or --NH--OR.sub.11, [0059] wherein R.sub.11
is H, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
aryl, heteroaryl, or heterocyclyl, and [0060] when Z is absent,
Ar.sub.1 is phenyl, R.sub.2 and R.sub.3 are H, n=1, and R.sub.4 is
--NHOH, then R.sub.1 is other than carbazole,
tetrahydro-.beta.-carboline, tetrahydro-.gamma.-carboline,
--C(.dbd.O)--NR.sub.5R.sub.6 and --NR.sub.5--C(.dbd.0)-R.sub.6,
wherein one of R.sub.5 or R.sub.6 is quinoline and the other of
R.sub.5 or R.sub.6 is H; or a pharmaceutically acceptable salt
thereof.
[0061] In some embodiments, the compound wherein Z is present and
is --O--, --S--, --CH.sub.2--, --NH--, --C(O)--, --NH--NH--,
--NHC(.dbd.O)--, --C(.dbd.O)NH--, --NHC(.dbd.O)CH.sub.2NH-- or
--NHC(.dbd.O)CH.dbd.CH--,
or a pharmaceutically acceptable salt thereof.
[0062] In some embodiments, the compound wherein Z is present and
is --O--, --S--, --NH--, --(CO)--, --NH--NH--, --NHC(.dbd.O)--,
--C(.dbd.O)NH--, --NHC(.dbd.O)CH.sub.2NH-- or
--NHC(.dbd.O)CH.dbd.CH--,
or a pharmaceutically acceptable salt thereof.
[0063] In some embodiments, the compound wherein Z is --NH--,
--NHC(.dbd.O), --NHC(.dbd.O)CH.sub.2NH-- or
--NHC(.dbd.O)CH.dbd.CH--,
or a pharmaceutically acceptable salt thereof.
[0064] In some embodiments, the compound wherein
##STR00005## [0065] wherein X is a Cl, Br, or F, or a
pharmaceutically acceptable salt thereof.
[0066] In some embodiments, the compound wherein [0067] Ar.sub.1
is
[0067] ##STR00006## [0068] wherein X is Cl, Br, or F, or a
pharmaceutically acceptable salt thereof.
[0069] In some embodiments, the compound wherein R.sub.1 is
--C(.dbd.O)--NR.sub.5R.sub.6 or --NR.sub.5--C(.dbd.0)-R.sub.6,
[0070] wherein [0071] R.sub.5 is alkyl, heteroalkyl, cycloalkyl,
aryl or heteroaryl; and [0072] R.sub.6 is alkyl, heteroalkyl,
cycloalkyl, aryl or heteroaryl, or a pharmaceutically acceptable
salt thereof.
[0073] In some embodiments, the compound [0074] wherein R.sub.5
is
[0074] ##STR00007## [0075] wherein [0076] X is a Cl, Br, or F;
[0077] R.sub.12 is H, C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5
alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or
heteroaryl; and [0078] R.sub.6 is H,
##STR00008##
[0078] or a pharmaceutically acceptable salt thereof.
[0079] In some embodiments, the compound [0080] wherein [0081] m=1;
[0082] R.sub.2 is H or CH.sub.3; and [0083] R.sub.3 is H, CH.sub.3,
Cl, Br, F, or CF.sub.3; or a pharmaceutically acceptable salt
thereof.
[0084] In some embodiments, the compound wherein R.sub.6 is
--NH--OR.sub.11 or --OR.sub.11, wherein R.sub.11 is H or
CH.sub.3,
or a pharmaceutically acceptable salt thereof.
[0085] In some embodiments, the compound wherein Z is absent,
or a pharmaceutically acceptable salt thereof.
[0086] In some embodiments, the compound
[0087] wherein Ar.sub.1 is
##STR00009## [0088] wherein X is a Cl, Br, or F, or a
pharmaceutically acceptable salt thereof.
[0089] In some embodiments, the compound wherein [0090] Ar.sub.1
is
[0090] ##STR00010## [0091] wherein X is Cl, Br, or F, or a
pharmaceutically acceptable salt thereof.
[0092] In some embodiments, the compound [0093] wherein R.sub.1 is
--C(.dbd.O)--NR.sub.5R.sub.6 or --NR.sub.5--C(.dbd.0)-R.sub.6,
[0094] wherein [0095] R.sub.5 is alkyl, heteroalkyl cycloalkyl,
aryl or heteroaryl; and [0096] R.sub.6 is alkyl, heteroalkyl,
cycloalkyl, aryl or heteroaryl, or a pharmaceutically acceptable
salt thereof.
[0097] In some embodiments, the compound [0098] wherein R.sub.1 is
--C(.dbd.O)--NR.sub.5R.sub.6, [0099] wherein [0100] R.sub.5 is
[0100] ##STR00011## [0101] wherein [0102] X is a Cl, Br, or F;
[0103] R.sub.12 is H, C.sub.1-5 alkyl, C.sub.2-5 alkenyl, C.sub.2-5
alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, or
heteroaryl; and [0104] R.sub.6 is H,
##STR00012##
[0104] or a pharmaceutically acceptable salt thereof.
[0105] In some embodiments, the compound [0106] wherein m=1; [0107]
R.sub.2 is H or CH.sub.3; and [0108] R.sub.3 is H, CH.sub.3, Cl,
Br, F, or CF.sub.3; or a pharmaceutically acceptable salt
thereof.
[0109] In some embodiments, the compound [0110] wherein R.sub.6 is
--NH--OR.sub.11 or --OR.sub.11, [0111] wherein R.sub.11 is H or
CH.sub.3, or a pharmaceutically acceptable salt thereof.
[0112] In some embodiments, the compound [0113] wherein R.sub.1 is
--C(.dbd.O)--NR.sub.5R.sub.6, [0114] wherein R.sub.5 and R.sub.6
heteroalkyl, aryl, or heteroaryl; [0115] m is 1; [0116] R.sub.2 and
R.sub.3 are H; [0117] Ar.sub.1 is phenyl; [0118] Z is absent;
[0119] R.sub.4 is --NH--OR.sub.11, [0120] wherein R.sub.11 is H, or
a pharmaceutically acceptable salt thereof.
[0121] In some embodiments, the compound having the structure:
##STR00013## ##STR00014## ##STR00015## ##STR00016##
or a pharmaceutically acceptable salt thereof.
[0122] In some embodiments, the compound having the structure:
##STR00017##
or a pharmaceutically acceptable salt thereof.
[0123] In some embodiments, the compound having the structure:
##STR00018##
or a pharmaceutically acceptable salt thereof.
[0124] In some embodiments of the compound, n=1; R.sub.1.dbd.H,
CH.sub.3, OH, NH.sub.2 or F; and R.sub.2 and R.sub.3=
##STR00019##
[0125] In some embodiments of the compound, n=0; and R.sub.1=
##STR00020##
[0126] In some embodiments of the compound, n=1; R.sub.1.dbd.H;
R.sub.2=
##STR00021##
and R.sub.3.dbd.CH.sub.3, OH, NH.sub.2 or F.
[0127] In some embodiments of the compound, n=1; R.sub.1.dbd.H,
CH.sub.3, OH, NH.sub.2 or F; and R.sub.2 and R.sub.3=
##STR00022##
[0128] In some embodiments of the compound, n=1;
R.sub.1.dbd.R.sub.2=
##STR00023##
and R.sub.3.dbd.CH.sub.3, OH, NH.sub.2 or F.
[0129] A pharmaceutical composition comprising any one, or more, of
the instant compounds and a pharmaceutically acceptable
carrier.
[0130] In some embodiments, the compound having the structure:
##STR00024## ##STR00025## ##STR00026## ##STR00027## [0131] wherein
Z is present or absent, and when present is --O--, --S--,
--CH.sub.2--, --NH--, --NH--NH--, --NHC(.dbd.O)--, --C(.dbd.O)NH--,
--NHC(.dbd.O)CH.sub.2NH-- or --NHC(.dbd.O)CH.dbd.CH--; and [0132]
R.sub.4 is --OR.sub.11 or --NH--OR.sub.11, [0133] wherein R.sub.11
is H, C.sub.1-10 alkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl,
aryl, heteroaryl, or heterocyclyl, or a pharmaceutically acceptable
salt thereof.
[0134] In some embodiments, the compound having the structure
##STR00028## [0135] wherein Z is present or absent, and when
present is --O--, --S--, --CH.sub.2--, --NH--, --NH--NH--,
--NHC(.dbd.O)--, --C(.dbd.O)NH--, --NHC(.dbd.O)CH.sub.2NH-- or
--NHC(.dbd.O)CH.dbd.CH--; and [0136] R.sub.4 is --OR.sub.11 or
--NH--OR.sub.11, [0137] wherein R.sub.11 is H, C.sub.1-10 alkyl,
C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, aryl, heteroaryl, or
heterocyclyl, or a pharmaceutically acceptable salt thereof.
[0138] In some embodiments, the compound wherein [0139] Ar.sub.1 is
indole, benzimidazole, or tetralin, or a pharmaceutically
acceptable salt thereof.
[0140] In some embodiments, the compound having the structure:
##STR00029## ##STR00030## ##STR00031##
or a pharmaceutically acceptable salt thereof.
[0141] In some embodiments, a pharmaceutical composition comprising
the compound of the present invention and a pharmaceutically
acceptable carrier.
[0142] In some embodiments, a method of inhibiting the activity of
a histone deactylase in a cell comprising contacting the histone
deacetylase with the compound or composition of any one of the
compounds of the present invention so as to inhibit the activity of
the histone deacetylase.
[0143] In some embodiments, a method of inhibiting the activity of
a histone deactylase wherein the histone deacetylase is HDAC6.
[0144] In some embodiments, a method of inhibiting the activity of
a histone deacetylase 6 (HDAC6) in a cell comprising contacting the
histone deacetylase 6 with the compound or composition of the
present invention so as to inhibit the activity of the histone
deacetylase 6 in the cell.
[0145] In some embodiments, a method of increasing accumulation of
acetylated alpha tubulin in a cell comprising contacting the cell
with any one of the compound or composition of the present
invention so as to increase the accumulation of acetylated
alpha-tubulin in the cell.
[0146] In some embodiments, a method of treating a
neurodegenerative disease in a subject comprising administering an
effective amount of the the compound or composition of the present
invention to the subject so as to treat the neurodegenerative
disease in the subject.
[0147] In some embodiments, a method of treating a
neurodegenerative disease wherein the neurodegenerative disease is
Parkinson's disease, Alzheimer's disease, Huntington's disease or
Niemann-Pick type C disease.
[0148] In some embodiments, a method of treating a disease
associated with defective lipid transport in a subject comprising
administering an effective amount of the compound or composition of
the present invention to the subject so as to treat the disease in
the subject.
[0149] In some embodiments, a method of treating a disease
associated with defective lipid transport wherein the disease
associated with defective lipid transport is Stargardt macular
degeneration, Harlequin ichthyosis or Tangier disease.
[0150] A method of inhibiting the activity of a histone deactylase
in a cell comprising contacting the histone deacetylase with any
one, or more, of the instant compounds so as to inhibit the
activity of the histone deacetylase.
[0151] In an embodiment the histone deacetylase is HDAC6.
[0152] A method of inhibiting the activity of a histone deacetylase
6 (HDAC6) in a cell comprising contacting the histone deacetylase 6
with any one, or more, of the instant compounds so as to inhibit
the activity of the histone deacetylase 6 in the cell.
[0153] A method of increasing accumulation of acetylated alpha
tubulin in a cell comprising contacting the cell with any one, or
more, of the instant compounds so as to increase the accumulation
of acetylated alpha-tubulin in the cell.
[0154] This invention also provides isotopic variants of the
compounds disclosed herein, including wherein the isotopic atom is
.sup.2H and/or wherein the isotopic atom .sup.13C. Accordingly, in
the compounds provided herein hydrogen can be enriched in the
deuterium isotope. It is to be understood that the invention
encompasses all such isotopic forms which inhibit HDAC, including
those which inhibit HDAC6 selectively over HDAC1.
[0155] A method of treating a neurodegenerative disease in a
subject comprising administering an effective amount of any one, or
more, of the instant compounds to the subject so as to treat the
neurodegenerative disease in the subject.
[0156] In an embodiment, the neurodegenerative disease is
Parkinson's disease, Alzheimer's disease, and Huntington's disease
or Niemann-Pick type C disease.
[0157] A method of treating a disease associated with defective
lipid transport in a subject comprising administering an effective
amount of any one, or more, of the instant compounds to the subject
so as to treat the disease in the subject.
[0158] In an embodiment, the disease associated with defective
lipid transport is Stargardt macular degeneration, Harlequin
ichthyosis or Tangier disease.
[0159] In some embodiments, the compounds of the instant invention
for use in the treatment of cancer.
[0160] In some embodiments, the compounds of the instant invention
for use in the treatment of a neurodegenerative disease.
[0161] In some embodiments, the compounds of the instant invention
for use in the treatment of a disease associated with defective
lipid transport.
[0162] In some embodiments, the compounds of the instant invention
for use in inhibiting the activity of a histone deactylase in a
cell.
[0163] In some embodiments, the compounds of the instant invention
for use in inhibiting the activity of histone deactylase 6 in a
cell.
[0164] In some embodiments, the compounds of the instant invention
for use in increasing accumulation of acetylated alpha tubulin in a
cell.
[0165] It is understood that the structures described in the
embodiments of the methods hereinabove can be the same as the
structures of the compounds described hereinabove.
[0166] It is understood that where a numerical range is recited
herein, the present invention contemplates each integer between,
and including, the upper and lower limits, unless otherwise
stated.
[0167] As used herein, the term "activity" refers to the
activation, production, expression, synthesis, intercellular
effect, and/or pathological or aberrant effect of the referenced
molecule, either inside and/or outside of a cell. Such molecules
include, but are not limited to, cytokines, enzymes, growth
factors, pro-growth factors, active growth factors, and
pro-enzymes. Molecules such as cytokines, enzymes, growth factors,
pro-growth factors, active growth factors, and pro-enzymes may be
produced, expressed, or synthesized within a cell where they may
exert an effect. Such molecules may also be transported outside of
the cell to the extracellular matrix where they may induce an
effect on the extracellular matrix or on a neighboring cell. It is
understood that activation of inactive cytokines, enzymes and
pro-enzymes may occur inside and/or outside of a cell and that both
inactive and active forms may be present at any point inside and/or
outside of a cell. It is also understood that cells may possess
basal levels of such molecules for normal function and that
abnormally high or low levels of such active molecules may lead to
pathological or aberrant effects that may be corrected by
pharmacological intervention.
[0168] As used herein, the term "histone deacetylase" or "HDAC"
refers to any member of the classes of enzymes capable of cleaving
an acetyl group (--C(.dbd.O)CH.sub.3) from proteins, which include,
but are not limited to, histones and microtubules. A histone
deacetylase may be zinc-dependent. Examples of HDACs include, but
are not limited to, HDAC1, HDAC2, HDAC3, HDAC4, HDAC5, HDAC6,
HDAC7, HDAC8, HDAC9, HDAC10, and HDAC11.
[0169] Except where otherwise specified, the structure of a
compound of this invention includes an asymmetric carbon atom, it
is understood that the compound occurs as a racemate, racemic
mixture, and isolated single enantiomer. All such isomeric forms of
these compounds are expressly included in this invention. Except
where otherwise specified, each stereogenic carbon may be of the R
or S configuration. It is to be understood accordingly that the
isomers arising from such asymmetry (e.g., all enantiomers and
diastereomers) are included within the scope of this invention,
unless indicated otherwise. Such isomers can be obtained in
substantially pure form by classical separation techniques and by
stereochemically controlled synthesis, such as those described in
"Enantiomers, Racemates and Resolutions" by J. Jacques, A. Collet
and S. Wilen, Pub. John Wiley & Sons, NY, 1981. For example,
the resolution may be carried out by preparative chromatography on
a chiral column.
[0170] The subject invention is also intended to include all
isotopes of atoms occurring on the compounds disclosed herein.
Isotopes include those atoms having the same atomic number but
different mass numbers. By way of general example and without
limitation, isotopes of hydrogen include tritium and deuterium.
Isotopes of carbon include C-13 and C-14.
[0171] It will be noted that any notation of a carbon in structures
throughout this application, when used without further notation,
are intended to represent all isotopes of carbon, such as .sup.12C,
.sup.13C, or .sup.14C. Furthermore, any compounds containing
.sup.13C or .sup.14C may specifically have the structure of any of
the compounds disclosed herein.
[0172] It will also be noted that any notation of a hydrogen in
structures throughout this application, when used without further
notation, are intended to represent all isotopes of hydrogen, such
as .sup.1H, .sup.2H, or .sup.3H. Furthermore, any compounds
containing .sup.2H or .sup.3H may specifically have the structure
of any of the compounds disclosed herein.
[0173] Isotopically-labeled compounds can generally be prepared by
conventional techniques known to those skilled in the art using
appropriate isotopically-labeled reagents in place of the
non-labeled reagents employed.
[0174] In the compounds used in the method of the present
invention, the substituents may be substituted or unsubstituted,
unless specifically defined otherwise.
[0175] In the compounds used in the method of the present
invention, alkyl, heteroalkyl, monocycle, bicycle, aryl, heteroaryl
and heterocycle groups can be further substituted by replacing one
or more hydrogen atoms with alternative non-hydrogen groups. These
include, but are not limited to, halo, hydroxy, mercapto, amino,
carboxy, cyano and carbamoyl.
[0176] It is understood that substituents and substitution patterns
on the compounds used in the method of the present invention can be
selected by one of ordinary skill in the art to provide compounds
that are chemically stable and that can be readily synthesized by
techniques known in the art from readily available starting
materials. If a substituent is itself substituted with more than
one group, it is understood that these multiple groups may be on
the same carbon or on different carbons, so long as a stable
structure results.
[0177] In choosing the compounds used in the method of the present
invention, one of ordinary skill in the art will recognize that the
various substituents, i.e. R.sub.1, R.sub.2, etc. are to be chosen
in conformity with well-known principles of chemical structure
connectivity.
[0178] As used herein, "alkyl" includes both branched and
straight-chain saturated aliphatic hydrocarbon groups having the
specified number of carbon atoms and may be unsubstituted or
substituted. Thus, C.sub.1-C.sub.n as in "C.sub.1-C.sub.n alkyl" is
defined to include groups having 1, 2, . . . , n-1 or n carbons in
a linear or branched arrangement. For example, C.sub.1-C.sub.6, as
in "C.sub.1-C.sub.6 alkyl" is defined to include groups having 1,
2, 3, 4, 5, or 6 carbons in a linear or branched arrangement, and
specifically includes methyl, ethyl, n-propyl, isopropyl, n-butyl,
t-butyl, pentyl, hexyl, and octyl.
[0179] As used herein, "alkenyl" refers to a non-aromatic
hydrocarbon radical, straight or branched, containing at least 1
carbon to carbon double bond, and up to the maximum possible number
of non-aromatic carbon-carbon double bonds may be present, and may
be unsubstituted or substituted. For example, "C.sub.2-C.sub.6
alkenyl" means an alkenyl radical having 2, 3, 4, 5, or 6 carbon
atoms, and up to 1, 2, 3, 4, or 5 carbon-carbon double bonds
respectively. Alkenyl groups include ethenyl, propenyl, butenyl and
cyclohexenyl.
[0180] The term "alkynyl" refers to a hydrocarbon radical straight
or branched, containing at least 1 carbon to carbon triple bond,
and up to the maximum possible number of non-aromatic carbon-carbon
triple bonds may be present, and may be unsubstituted or
substituted. Thus, "C.sub.2-C.sub.6 alkynyl" means an alkynyl
radical having 2 or 3 carbon atoms and 1 carbon-carbon triple bond,
or having 4 or 5 carbon atoms and up to 2 carbon-carbon triple
bonds, or having 6 carbon atoms and up to 3 carbon-carbon triple
bonds. Alkynyl groups include ethynyl, propynyl and butynyl.
[0181] "Alkylene", "alkenylene" and "alkynylene" shall mean,
respectively, a divalent alkane, alkene and alkyne radical,
respectively. It is understood that an alkylene, alkenylene, and
alkynylene may be straight or branched. An alkylene, alkenylene,
and alkynylene may be unsubstituted or substituted.
[0182] As used herein, "aryl" is intended to mean any stable
monocyclic, bicyclic or polycyclic carbon ring of up to 10 atoms in
each ring, wherein at least one ring is aromatic, and may be
unsubstituted or substituted. Examples of such aryl elements
include phenyl, p-toluenyl (4-methylphenyl), naphthyl,
tetrahydro-naphthyl, indanyl, biphenyl, phenanthryl, anthryl or
acenaphthyl. In cases where the aryl substituent is bicyclic and
one ring is non-aromatic, it is understood that attachment is via
the aromatic ring.
[0183] As used herein, the term "polycyclic" refers to unsaturated
or partially unsaturated multiple fused ring structures, which may
be unsubstituted or substituted.
[0184] The term "arylalkyl" refers to alkyl groups as described
above wherein one or more bonds to hydrogen contained therein are
replaced by a bond to an aryl group as described above. It is
understood that an "arylalkyl" group is connected to a core
molecule through a bond from the alkyl group and that the aryl
group acts as a substituent on the alkyl group. Examples of
arylalkyl moieties include, but are not limited to, benzyl
(phenylmethyl), p-trifluoromethylbenzyl
(4-trifluoromethylphenylmethyl), 1-phenylethyl, 2-phenylethyl,
3-phenylpropyl, 2-phenylpropyl and the like.
[0185] The term "heteroaryl", as used herein, represents a stable
monocyclic, bicyclic or polycyclic ring of up to 10 atoms in each
ring, wherein at least one ring is aromatic and contains from 1 to
4 heteroatoms selected from the group consisting of O, N and S.
Bicyclic aromatic heteroaryl groups include phenyl, pyridine,
pyrimidine or pyridizine rings that are (a) fused to a 6-membered
aromatic (unsaturated) heterocyclic ring having one nitrogen atom;
(b) fused to a 5- or 6-membered aromatic (unsaturated) heterocyclic
ring having two nitrogen atoms; (c) fused to a 5-membered aromatic
(unsaturated) heterocyclic ring having one nitrogen atom together
with either one oxygen or one sulfur atom; or (d) fused to a
5-membered aromatic (unsaturated) heterocyclic ring having one
heteroatom selected from O, N or S. Heteroaryl groups within the
scope of this definition include but are not limited to:
benzoimidazolyl, benzofuranyl, benzofurazanyl, benzopyrazolyl,
benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazolyl,
carbolinyl, cinnolinyl, furanyl, indolinyl, indolyl, indolazinyl,
indazolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl,
isoxazolyl, naphthpyridinyl, oxadiazolyl, oxazolyl, oxazoline,
isoxazoline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl,
pyridopyridinyl, pyridazinyl, pyridyl, pyrimidyl, pyrrolyl,
quinazolinyl, quinolyl, quinoxalinyl, tetrazolyl, tetrazolopyridyl,
thiadiazolyl, thiazolyl, thienyl, triazolyl, azetidinyl,
aziridinyl, 1,4-dioxanyl, hexahydroazepinyl,
dihydrobenzoimidazolyl, dihydrobenzofuranyl,
dihydrobenzothiophenyl, dihydrobenzoxazolyl, dihydrofuranyl,
dihydroimidazolyl, dihydroindolyl, dihydroisooxazolyl,
dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl,
dihydropyrazinyl, dihydropyrazolyl, dihydropyridinyl,
dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl,
dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl,
dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl,
methylenedioxybenzoyl, tetrahydrofuranyl, tetrahydrothienyl,
acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrrazolyl,
indolyl, benzotriazolyl, benzothiazolyl, benzoxazolyl, isoxazolyl,
isothiazolyl, furanyl, thienyl, benzothienyl, benzofuranyl,
quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl,
pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl,
tetra-hydroquinoline. In cases where the heteroaryl substituent is
bicyclic and one ring is non-aromatic or contains no heteroatoms,
it is understood that attachment is via the aromatic ring or via
the heteroatom containing ring, respectively. If the heteroaryl
contains nitrogen atoms, it is understood that the corresponding
N-oxides thereof are also encompassed by this definition.
[0186] The term "heterocycle", "heterocyclyl" or "heterocyclic"
refers to a mono- or poly-cyclic ring system which can be saturated
or contains one or more degrees of unsaturation and contains one or
more heteroatoms. Preferred heteroatoms include N, O, and/or S,
including N-oxides, sulfur oxides, and dioxides. Preferably the
ring is three to ten-membered and is either saturated or has one or
more degrees of unsaturation. The heterocycle may be unsubstituted
or substituted, with multiple degrees of substitution being
allowed. Such rings may be optionally fused to one or more of
another "heterocyclic" ring(s), heteroaryl ring(s), aryl ring(s),
or cycloalkyl ring(s). Examples of heterocycles include, but are
not limited to, tetrahydrofuran, pyran, 1,4-dioxane, 1,3-dioxane,
piperidine, piperazine, pyrrolidine, morpholine, thiomorpholine,
tetrahydrothiopyran, tetrahydrothiophene, 1,3-oxathiolane, and the
like.
[0187] The alkyl, alkenyl, alkynyl, aryl, heteroaryl and
heterocyclyl substituents may be substituted or unsubstituted,
unless specifically defined otherwise.
[0188] In the compounds of the present invention, alkyl, alkenyl,
alkynyl, aryl, heterocyclyl and heteroaryl groups can be further
substituted by replacing one or more hydrogen atoms with
alternative non-hydrogen groups. These include, but are not limited
to, halo, hydroxy, mercapto, amino, carboxy, cyano and
carbamoyl.
[0189] As used herein, the term "halogen" refers to F, Cl, Br, and
I.
[0190] As used herein, "heteroalkyl" includes both branched and
straight-chain saturated aliphatic hydrocarbon groups having the
specified number of carbon atoms and at least 1 heteroatom within
the chain or branch.
[0191] As used herein, "monocycle" includes any stable polyatomic
carbon ring of up to 10 atoms and may be unsubstituted or
substituted. Examples of such non-aromatic monocycle elements
include but are not limited to: cyclobutyl, cyclopentyl,
cyclohexyl, and cycloheptyl. Examples of such aromatic monocycle
elements include but are not limited to: phenyl.
[0192] As used herein, "bicycle" includes any stable polyatomic
carbon ring of up to 10 atoms that is fused to a polyatomic carbon
ring of up to 10 atoms with each ring being independently
unsubstituted or substituted. Examples of such non-aromatic bicycle
elements include but are not limited to: decahydronaphthalene.
Examples of such aromatic bicycle elements include but are not
limited to: naphthalene.
[0193] The term "ester" is intended to a mean an organic compound
containing the R--O--CO--R' group.
[0194] The term "amide" is intended to a mean an organic compound
containing the R--CO--NH--R' or R--CO--N--R'R'' group.
[0195] The term "phenyl" is intended to mean an aromatic six
membered ring containing six carbons.
[0196] The term "biphenyl" is intended to mean an aryl comprising
two benzene rings linked together, and any substituted derivative
thereof.
[0197] The term "pyrrole" is intended to mean a heteroaryl having a
five-membered ring containing four carbon atoms and one nitrogen
atom.
[0198] The term "furan" is intended to mean a heteroaryl having a
five-membered ring containing four carbon atoms and one oxygen
atom.
[0199] The term "thiophene" is intended to mean a heteroaryl having
a five-membered ring containing four carbon atoms and one sulfur
atom.
[0200] The term "quinoline" is intended to mean a fully aromatic
heteroaryl having a six-membered ring fused to a six-membered ring
containing nine carbon atoms and one nitrogen atom.
[0201] The term "carbazole" is intended to mean a fully aromatic
heteroaryl having two six-membered benzene rings fused both sides
of a five-membered nitrogen-containing ring.
[0202] The terms "tetrahydrocarboline",
"tetrahydro-.beta.-carboline" and "tetrahydro-.gamma.-carboline"
are intended to mean an aryl having a six-membered ring fused to a
five membered ring which in turn is fused to a piperidine ring with
a total of 11 carbon atoms and 2 nitrogen atoms with the
five-membered ring containing 1 nitrogen atom and the piperidine
ring containing 1 nitrogen atom and any substituted derivative
thereof.
[0203] The term "benzimidazole" is intended to mean a heteroaryl
having a five-membered ring fused to a phenyl ring with the
five-membered ring containing 2 nitrogen atoms directly attached to
the phenyl ring.
[0204] The term "indole" is intended to mean a heteroaryl having a
five-membered ring fused to a phenyl ring with the five-membered
ring containing 1 nitrogen atom directly attached to the phenyl
ring.
[0205] The term "tetralin" is intended to mean a bicycle wherein an
aromatic phenyl ring is fused to a non-aromatic cyclohexyl
ring.
[0206] The term "substitution", "substituted" and "substituent"
refers to a functional group as described above in which one or
more bonds to a hydrogen atom contained therein are replaced by a
bond to non-hydrogen or non-carbon atoms, provided that normal
valencies are maintained and that the substitution results in a
stable compound. Substituted groups also include groups in which
one or more bonds to a carbon(s) or hydrogen(s) atom are replaced
by one or more bonds, including double or triple bonds, to a
heteroatom. Examples of substituent groups include the functional
groups described above, and halogens (i.e., F, Cl, Br, and I);
alkyl groups, such as methyl, ethyl, n-propyl, isopropryl, n-butyl,
tert-butyl, and trifluoromethyl; hydroxyl; alkoxy groups, such as
methoxy, ethoxy, n-propoxy, and isopropoxy; aryloxy groups, such as
phenoxy; arylalkyloxy, such as benzyloxy (phenylmethoxy) and
p-trifluoromethylbenzyloxy (4-trifluoromethylphenylmethoxy);
heteroaryloxy groups; sulfonyl groups, such as
trifluoromethanesulfonyl, methanesulfonyl, and p-toluenesulfonyl;
nitro, nitrosyl; mercapto; sulfanyl groups, such as methylsulfanyl,
ethylsulfanyl and propylsulfanyl; cyano; amino groups, such as
amino, methylamino, dimethylamino, ethylamino, and diethylamino;
and carboxyl. Where multiple substituent moieties are disclosed or
claimed, the substituted compound can be independently substituted
by one or more of the disclosed or claimed substituent moieties,
singly or plurally. By independently substituted, it is meant that
the (two or more) substituents can be the same or different.
[0207] It is understood that substituents and substitution patterns
on the compounds of the instant invention can be selected by one of
ordinary skill in the art to provide compounds that are chemically
stable and that can be readily synthesized by techniques known in
the art, as well as those methods set forth below, from readily
available starting materials. If a substituent is itself
substituted with more than one group, it is understood that these
multiple groups may be on the same carbon or on different carbons,
so long as a stable structure results.
[0208] In choosing the compounds of the present invention, one of
ordinary skill in the art will recognize that the various
substituents, i.e. R.sub.1, R.sub.2, etc. are to be chosen in
conformity with well-known principles of chemical structure
connectivity.
[0209] The various R groups attached to the aromatic rings of the
compounds disclosed herein may be added to the rings by standard
procedures, for example those set forth in Advanced Organic
Chemistry: Part B: Reaction and Synthesis, Francis Carey and
Richard Sundberg, (Springer) 5th ed. Edition. (2007), the content
of which is hereby incorporated by reference.
[0210] The compounds used in the method of the present invention
may be prepared by techniques well know in organic synthesis and
familiar to a practitioner ordinarily skilled in the art. However,
these may not be the only means by which to synthesize or obtain
the desired compounds.
[0211] The compounds used in the method of the present invention
may be prepared by techniques described in Vogel's Textbook of
Practical Organic Chemistry, A. I. Vogel, A. R. Tatchell, B. S.
Furnis, A. J. Hannaford, P. W. G. Smith, (Prentice Hall) 5.sup.th
Edition (1996), March's Advanced Organic Chemistry: Reactions,
Mechanisms, and Structure, Michael B. Smith, Jerry March,
(Wiley-Interscience) 5.sup.th Edition (2007), and references
therein, which are incorporated by reference herein. However, these
may not be the only means by which to synthesize or obtain the
desired compounds.
[0212] Another aspect of the invention comprises a compound used in
the method of the present invention as a pharmaceutical
composition.
[0213] As used herein, the term "pharmaceutically active agent"
means any substance or compound suitable for administration to a
subject and furnishes biological activity or other direct effect in
the treatment, cure, mitigation, diagnosis, or prevention of
disease, or affects the structure or any function of the subject.
Pharmaceutically active agents include, but are not limited to,
substances and compounds described in the Physicians' Desk
Reference (PDR Network, LLC; 64th edition; Nov. 15, 2009) and
"Approved Drug Products with Therapeutic Equivalence Evaluations"
(U.S. Department Of Health And Human Services, 30.sup.th edition,
2010), which are hereby incorporated by reference. Pharmaceutically
active agents which have pendant carboxylic acid groups may be
modified in accordance with the present invention using standard
esterification reactions and methods readily available and known to
those having ordinary skill in the art of chemical synthesis. Where
a pharmaceutically active agent does not possess a carboxylic acid
group, the ordinarily skilled artisan will be able to design and
incorporate a carboxylic acid group into the pharmaceutically
active agent where esterification may subsequently be carried out
so long as the modification does not interfere with the
pharmaceutically active agent's biological activity or effect.
[0214] The compounds used in the method of the present invention
may be in a salt form. As used herein, a "salt" is a salt of the
instant compounds which has been modified by making acid or base
salts of the compounds. In the case of compounds used to treat an
infection or disease caused by a pathogen, the salt is
pharmaceutically acceptable. Examples of pharmaceutically
acceptable salts include, but are not limited to, mineral or
organic acid salts of basic residues such as amines; alkali or
organic salts of acidic residues such as phenols. The salts can be
made using an organic or inorganic acid. Such acid salts are
chlorides, bromides, sulfates, nitrates, phosphates, sulfonates,
formates, tartrates, maleates, malates, citrates, benzoates,
salicylates, ascorbates, and the like. Phenolate salts are the
alkaline earth metal salts, sodium, potassium or lithium. The term
"pharmaceutically acceptable salt" in this respect, refers to the
relatively non-toxic, inorganic and organic acid or base addition
salts of compounds of the present invention. These salts can be
prepared in situ during the final isolation and purification of the
compounds of the invention, or by separately reacting a purified
compound of the invention in its free base or free acid form with a
suitable organic or inorganic acid or base, and isolating the salt
thus formed. Representative salts include the hydrobromide,
hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate,
valerate, oleate, palmitate, stearate, laurate, benzoate, lactate,
phosphate, tosylate, citrate, maleate, fumarate, succinate,
tartrate, napthylate, mesylate, glucoheptonate, lactobionate, and
laurylsulphonate salts and the like. (See, e.g., Berge et al.
(1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).
[0215] As used herein, "treating" means preventing, slowing,
halting, or reversing the progression of a disease or infection.
Treating may also mean improving one or more symptoms of a disease
or infection.
[0216] The compounds used in the method of the present invention
may be administered in various forms, including those detailed
herein. The treatment with the compound may be a component of a
combination therapy or an adjunct therapy, i.e. the subject or
patient in need of the drug is treated or given another drug for
the disease in conjunction with one or more of the instant
compounds. This combination therapy can be sequential therapy where
the patient is treated first with one drug and then the other or
the two drugs are given simultaneously. These can be administered
independently by the same route or by two or more different routes
of administration depending on the dosage forms employed.
[0217] As used herein, a "pharmaceutically acceptable carrier" is a
pharmaceutically acceptable solvent, suspending agent or vehicle,
for delivering the instant compounds to the animal or human. The
carrier may be liquid or solid and is selected with the planned
manner of administration in mind. Liposomes are also a
pharmaceutically acceptable carrier.
[0218] The dosage of the compounds administered in treatment will
vary depending upon factors such as the pharmacodynamic
characteristics of a specific chemotherapeutic agent and its mode
and route of administration; the age, sex, metabolic rate,
absorptive efficiency, health and weight of the recipient; the
nature and extent of the symptoms; the kind of concurrent treatment
being administered; the frequency of treatment with; and the
desired therapeutic effect.
[0219] A dosage unit of the compounds used in the method of the
present invention may comprise a single compound or mixtures
thereof with additional antibacterial agents. The compounds can be
administered in oral dosage forms as tablets, capsules, pills,
powders, granules, elixirs, tinctures, suspensions, syrups, and
emulsions. The compounds may also be administered in intravenous
(bolus or infusion), intraperitoneal, subcutaneous, or
intramuscular form, or introduced directly, e.g. by injection,
topical application, or other methods, into or onto a site of
infection, all using dosage forms well known to those of ordinary
skill in the pharmaceutical arts.
[0220] The compounds used in the method of the present invention
can be administered in admixture with suitable pharmaceutical
diluents, extenders, excipients, or carriers (collectively referred
to herein as a pharmaceutically acceptable carrier) suitably
selected with respect to the intended form of administration and as
consistent with conventional pharmaceutical practices. The unit
will be in a form suitable for oral, rectal, topical, intravenous
or direct injection or parenteral administration. The compounds can
be administered alone or mixed with a pharmaceutically acceptable
carrier. This carrier can be a solid or liquid, and the type of
carrier is generally chosen based on the type of administration
being used. The active agent can be co-administered in the form of
a tablet or capsule, liposome, as an agglomerated powder or in a
liquid form. Examples of suitable solid carriers include lactose,
sucrose, gelatin and agar. Capsule or tablets can be easily
formulated and can be made easy to swallow or chew; other solid
forms include granules, and bulk powders. Tablets may contain
suitable binders, lubricants, diluents, disintegrating agents,
coloring agents, flavoring agents, flow-inducing agents, and
melting agents. Examples of suitable liquid dosage forms include
solutions or suspensions in water, pharmaceutically acceptable fats
and oils, alcohols or other organic solvents, including esters,
emulsions, syrups or elixirs, suspensions, solutions and/or
suspensions reconstituted from non-effervescent granules and
effervescent preparations reconstituted from effervescent granules.
Such liquid dosage forms may contain, for example, suitable
solvents, preservatives, emulsifying agents, suspending agents,
diluents, sweeteners, thickeners, and melting agents. Oral dosage
forms optionally contain flavorants and coloring agents. Parenteral
and intravenous forms may also include minerals and other materials
to make them compatible with the type of injection or delivery
system chosen.
[0221] Techniques and compositions for making dosage forms useful
in the present invention are described in the following references:
7 Modern Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes,
Editors, 1979); Pharmaceutical Dosage Forms: Tablets (Lieberman et
al., 1981); Ansel, Introduction to Pharmaceutical Dosage Forms 2nd
Edition (1976); Remington's Pharmaceutical Sciences, 17th ed. (Mack
Publishing Company, Easton, Pa., 1985); Advances in Pharmaceutical
Sciences (David Ganderton, Trevor Jones, Eds., 1992); Advances in
Pharmaceutical Sciences Vol. 7. (David Ganderton, Trevor Jones,
James McGinity, Eds., 1995); Aqueous Polymeric Coatings for
Pharmaceutical Dosage Forms (Drugs and the Pharmaceutical Sciences,
Series 36 (James McGinity, Ed., 1989); Pharmaceutical Particulate
Carriers: Therapeutic Applications: Drugs and the Pharmaceutical
Sciences, Vol 61 (Alain Rolland, Ed., 1993); Drug Delivery to the
Gastrointestinal Tract (Ellis Horwood Books in the Biological
Sciences. Series in Pharmaceutical Technology; J. G. Hardy, S. S.
Davis, Clive G. Wilson, Eds.); Modem Pharmaceutics Drugs and the
Pharmaceutical Sciences, Vol 40 (Gilbert S. Banker, Christopher T.
Rhodes, Eds.). All of the aforementioned publications are
incorporated by reference herein.
[0222] Tablets may contain suitable binders, lubricants,
disintegrating agents, coloring agents, flavoring agents,
flow-inducing agents, and melting agents. For instance, for oral
administration in the dosage unit form of a tablet or capsule, the
active drug component can be combined with an oral, non-toxic,
pharmaceutically acceptable, inert carrier such as lactose,
gelatin, agar, starch, sucrose, glucose, methyl cellulose,
magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol,
sorbitol and the like. Suitable binders include starch, gelatin,
natural sugars such as glucose or beta-lactose, corn sweeteners,
natural and synthetic gums such as acacia, tragacanth, or sodium
alginate, carboxymethylcellulose, polyethylene glycol, waxes, and
the like. Lubricants used in these dosage forms include sodium
oleate, sodium stearate, magnesium stearate, sodium benzoate,
sodium acetate, sodium chloride, and the like. Disintegrators
include, without limitation, starch, methyl cellulose, agar,
bentonite, xanthan gum, and the like.
[0223] The compounds used in the method of the present invention
may also be administered in the form of liposome delivery systems,
such as small unilamellar vesicles, large unilamallar vesicles, and
multilamellar vesicles. Liposomes can be formed from a variety of
phospholipids, such as cholesterol, stearylamine, or
phosphatidylcholines. The compounds may be administered as
components of tissue-targeted emulsions.
[0224] The compounds used in the method of the present invention
may also be coupled to soluble polymers as targetable drug carriers
or as a prodrug. Such polymers include polyvinylpyrrolidone, pyran
copolymer, polyhydroxylpropylmethacrylamide-phenol,
polyhydroxyethylasparta-midephenol, or polyethyleneoxide-polylysine
substituted with palmitoyl residues. Furthermore, the compounds may
be coupled to a class of biodegradable polymers useful in achieving
controlled release of a drug, for example, polylactic acid,
polyglycolic acid, copolymers of polylactic and polyglycolic acid,
polyepsilon caprolactone, polyhydroxy butyric acid,
polyorthoesters, polyacetals, polydihydropyrans, polycyanoacylates,
and crosslinked or amphipathic block copolymers of hydrogels.
[0225] Gelatin capsules may contain the active ingredient compounds
and powdered carriers, such as lactose, starch, cellulose
derivatives, magnesium stearate, stearic acid, and the like.
Similar diluents can be used to make compressed tablets. Both
tablets and capsules can be manufactured as immediate release
products or as sustained release products to provide for continuous
release of medication over a period of hours. Compressed tablets
can be sugar coated or film coated to mask any unpleasant taste and
protect the tablet from the atmosphere, or enteric coated for
selective disintegration in the gastrointestinal tract.
[0226] For oral administration in liquid dosage form, the oral drug
components are combined with any oral, non-toxic, pharmaceutically
acceptable inert carrier such as ethanol, glycerol, water, and the
like. Examples of suitable liquid dosage forms include solutions or
suspensions in water, pharmaceutically acceptable fats and oils,
alcohols or other organic solvents, including esters, emulsions,
syrups or elixirs, suspensions, solutions and/or suspensions
reconstituted from non-effervescent granules and effervescent
preparations reconstituted from effervescent granules. Such liquid
dosage forms may contain, for example, suitable solvents,
preservatives, emulsifying agents, suspending agents, diluents,
sweeteners, thickeners, and melting agents.
[0227] Liquid dosage forms for oral administration can contain
coloring and flavoring to increase patient acceptance. In general,
water, a suitable oil, saline, aqueous dextrose (glucose), and
related sugar solutions and glycols such as propylene glycol or
polyethylene glycols are suitable carriers for parenteral
solutions. Solutions for parenteral administration preferably
contain a water soluble salt of the active ingredient, suitable
stabilizing agents, and if necessary, buffer substances.
Antioxidizing agents such as sodium bisulfite, sodium sulfite, or
ascorbic acid, either alone or combined, are suitable stabilizing
agents. Also used are citric acid and its salts and sodium EDTA. In
addition, parenteral solutions can contain preservatives, such as
benzalkonium chloride, methyl- or propyl-paraben, and
chlorobutanol. Suitable pharmaceutical carriers are described in
Remington's Pharmaceutical Sciences, Mack Publishing Company, a
standard reference text in this field.
[0228] The compounds used in the method of the present invention
may also be administered in intranasal form via use of suitable
intranasal vehicles, or via transdermal routes, using those forms
of transdermal skin patches well known to those of ordinary skill
in that art. To be administered in the form of a transdermal
delivery system, the dosage administration will generally be
continuous rather than intermittent throughout the dosage
regimen.
[0229] Parenteral and intravenous forms may also include minerals
and other materials to make them compatible with the type of
injection or delivery system chosen.
[0230] The compounds and compositions of the present invention can
be administered in oral dosage forms as tablets, capsules, pills,
powders, granules, elixirs, tinctures, suspensions, syrups, and
emulsions. The compounds may also be administered in intravenous
(bolus or infusion), intraperitoneal, subcutaneous, or
intramuscular form, or introduced directly, e.g. by topical
administration, injection or other methods, to the afflicted area,
such as a wound, including ulcers of the skin, all using dosage
forms well known to those of ordinary skill in the pharmaceutical
arts.
[0231] Specific examples of pharmaceutical acceptable carriers and
excipients that may be used to formulate oral dosage forms of the
present invention are described in U.S. Pat. No. 3,903,297 to
Robert, issued Sep. 2, 1975. Techniques and compositions for making
dosage forms useful in the present invention are described-in the
following references: 7 Modern Pharmaceutics, Chapters 9 and 10
(Banker & Rhodes, Editors, 1979); Pharmaceutical Dosage Forms:
Tablets (Lieberman et al., 1981); Ansel, Introduction to
Pharmaceutical Dosage Forms 2nd Edition (1976); Remington's
Pharmaceutical Sciences, 17th ed. (Mack Publishing Company, Easton,
Pa., 1985); Advances in Pharmaceutical Sciences (David Ganderton,
Trevor Jones, Eds., 1992); Advances in Pharmaceutical Sciences Vol
7. (David Ganderton, Trevor Jones, James McGinity, Eds., 1995);
Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms (Drugs
and the Pharmaceutical Sciences, Series 36 (James McGinity, Ed.,
1989); Pharmaceutical Particulate Carriers: Therapeutic
Applications: Drugs and the Pharmaceutical Sciences, Vol 61 (Alain
Rolland, Ed., 1993); Drug Delivery to the Gastrointestinal Tract
(Ellis Horwood Books in the Biological Sciences. Series in
Pharmaceutical Technology; J. G. Hardy, S. S. Davis, Clive G.
Wilson, Eds.); Modem Pharmaceutics Drugs and the Pharmaceutical
Sciences, Vol 40 (Gilbert S. Banker, Christopher T. Rhodes, Eds.).
All of the aforementioned publications are incorporated by
reference herein.
[0232] The term "prodrug" as used herein refers to any compound
that when administered to a biological system generates the
compound of the invention, as a result of spontaneous chemical
reaction(s), enzyme catalyzed chemical reaction(s), photolysis,
and/or metabolic chemical reaction(s). A prodrug is thus a
covalently modified analog or latent form of a compound of the
invention.
[0233] The active ingredient can be administered orally in solid
dosage forms, such as capsules, tablets, powders, and chewing gum;
or in liquid dosage forms, such as elixirs, syrups, and
suspensions, including, but not limited to, mouthwash and
toothpaste. It can also be administered parentally, in sterile
liquid dosage forms.
[0234] Solid dosage forms, such as capsules and tablets, may be
enteric coated to prevent release of the active ingredient
compounds before they reach the small intestine. Materials that may
be used as enteric coatings include, but are not limited to,
sugars, fatty acids, waxes, shellac, cellulose acetate phthalate
(CAP), methyl acrylate-methacrylic acid copolymers, cellulose
acetate succinate, hydroxy propyl methyl cellulose phthalate,
hydroxy propyl methyl cellulose acetate succinate (hypromellose
acetate succinate), polyvinyl acetate phthalate (PVAP), and methyl
methacrylate-methacrylic acid copolymers.
[0235] The compounds and compositions of the invention can be
coated onto stents for temporary or permanent implantation into the
cardiovascular system of a subject.
[0236] The compounds of the present invention can be synthesized
according to general Schemes. Variations on the following general
synthetic methods will be readily apparent to those skilled in the
art and are deemed to be within the scope of the present
invention.
[0237] Each embodiment disclosed herein is contemplated as being
applicable to each of the other disclosed embodiments. Thus, all
combinations of the various elements described herein are within
the scope of the invention.
[0238] This invention will be better understood by reference to the
Experimental Details which follow, but those skilled in the art
will readily appreciate that the specific experiments detailed are
only illustrative of the invention as described more fully in the
claims which follow thereafter.
EXPERIMENTAL DETAILS
[0239] 8-Aminoquinoline, aniline, glycolaldehyde dimer, sodium
triacetoxyborohydride, tert-butylchlorodimethylsilane (TBDMS-Cl),
potassium cyanide, NH.sub.2OH, trifluoroacetic acid (TFA),
dichloroethane (DCE), dichloromethne (DCM),
1-Ethyl-3-(3-dimethyllaminopropyl)carbodiimide hydrochloride (EDC),
sodium sulfate (anhydrous), calcium hypochlorite, molecular sieves
(4 .ANG.), NH.sub.4Cl, NaCl, MeOH, NaHCO.sub.3, THF, hydrochloric
acid, acetic acid, CDCl.sub.3, CD.sub.3OD, and hexanes were used as
received without further purification. Purification of product
mixtures was carried out by column using silica gel with 40-60
.ANG. particle size or preparative chromatography using silica gel
60F 254 TLC-plates. TLC was carried out using silica gel 60F 254
TLC-plates. Proton NMR data were acquired at 400 MHz and .sup.13C
NMR data were acquired at 100.6 MHz.
[0240] Those having ordinary skill in the art of organic synthesis
will appreciate that modifications to general procedures and
synthetic routes contained in this application can be used to yield
additional derivatives and structurally diverse compounds. Suitable
organic transformations are described in in March's Advanced
Organic Chemistry: Reactions, Mechanisms, and Structure
(Wiley-Interscience; 6.sup.th edition, 2007), the content of which
is hereby incorporated by reference.
Example 1
Synthesis of Compound 6
[0241] HDAC inhibitor 6 was accessed according to the protocols
shown in Scheme 1. Alcohol 2 was prepared by treatment of aniline 1
with glycolaldehyde followed by addition of sodium
triacetoxyborohydride. The alcohol 2 was then protected with
TBDMSCl followed by EDC coupling with acid 8 (Scheme 2). The methyl
ester 4 was converted directly, using a catalytic amount of KCN, to
the corresponding hydroxamic acid 5, which was deprotected with TFA
to afford compound 6.
##STR00032##
[0242] Reagents: (a) 1. HOCH.sub.2CHO, DCE, 30 min. rt. 2.
NaBH(OAc).sub.3, 4 h. rt. (b) TBDMS-Cl, DCM, 3 h. (c) 8, EDC, DCM,
rt. 24 h. (d) NH.sub.2OH, THF:MeOH (1:1), KCN (cat.) 16 h, rt. (e)
5% TFA in DCM, 5 min
##STR00033##
[0243] Reagents: (a) Ca(OCl).sub.2, HOAc, ACN, MeOH, molecular
sieves, dark, 48 hr, rt
2-(phenylamino)ethanol (2)
[0244] Glycolaldehyde (645 mg, 10.7 mmol) was added to a solution
containing aniline (1, 1.00 g, 10.7 mmol) in DCE (30 mL). The
solution was stirred at room temperature in argon atmosphere for 30
min. Sodium triacetoxyborohydride (2.61 g, 12.3 mmol) was then
added portion-wise. The reaction mixture was further stirred for 4
h before and quenched with NaHCO.sub.3 to pH=9.0. The reaction
mixture was extracted with mixed solvent (CHCl.sub.3:i-PrOH=4:1, 30
mL.times.3). The organic layer was dried (anhydrous sodium sulfate)
and concentrated in vacuo. The crude product was chromatographed on
silica gel (CH.sub.2Cl.sub.2/MeOH, 20:1) to yield target compound
2. Yield 1.061 g, 72%. R.sub.f=0.45, .sup.1H NMR (CHCl.sub.3, 400
MHz): .delta. 7.24 (dd, J=8.8, 7.4 Hz, 2H), 6.80 (t, J=7.4 Hz, 1H),
6.69 (d, J=8.8, 2H), 3.81 (t, J=5.2 Hz, 2H), 3.30 (t, J=5.2 Hz,
2H), 2.6 (br s, 1H); .sup.13C NMR (CHCl.sub.3, 100 MHz): .delta.
148.2, 129.4, 118.0, 113.3, 61.2, 46.1; [M+H].sup.+=138.08
(APCI+).
N-(2-((tert-butyldimethylsilyl)oxy)ethyl)aniline (3)
[0245] TBDMS-Cl (1.28 g, 8.02 mmol) and imidazole (1.45 g, 21.86
mmol) was added to a solution containing 2-(phenylamino)ethanol 2,
(1.00 g, 7.29 mmol) in CH.sub.2Cl.sub.2 (10 mL). The reaction
mixture was stirred at room temperature in argon atmosphere for 3
h. Then the reaction was quenched with sat. NH.sub.4Cl. and washed
with water (10 mL.times.2) and brine (10 mL.times.2). The organic
layer was dried (anhydrous sodium sulfate) and concentrated in
vacuo. The crude product was chromatographed on silica gel
(Hexanes/EtOAc, 7:1) to yield target compound 3. Yield 1.689 g,
92%. R.sub.f=0.60, .sup.1H NMR (CHCl.sub.3, 400 MHz): .delta. 7.22
(dd, J=8.8, 7.4 Hz, 2H), 6.76 (t, J=7.4 Hz, 1H), 6.68 (d, J=8.8,
2H), 4.09 (br s, 1H), 3.86 (t, J=5.2 Hz, 2H), 3.26 (t, J=5.2 Hz,
2H), 0.95 (s, 9H), 0.11 (s, 6H); .sup.13C NMR (CHCl.sub.3, 100
MHz): .delta. 148.4, 129.2, 117.5, 113.2, 61.6, 46.0, 25.9, 18.3,
-5.3; [M+H].sup.+=252.12 (APCI+).
Methyl 4-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)(phenyl)
amino)-2-oxoethyl) benzoate (4)
[0246] EDC (114.5 mg, 0.597 mmol) was added to a solution
containing N-(2-((tert-butyldimethylsilyl)oxy)ethyl)aniline 3 (100
mg, 0.398 mmol) and 2-(4-(methoxycarbonyl)phenyl)acetic acid, 8,
(116 mg, 0.597 mmol) in CH.sub.2Cl.sub.2 (3 mL). The reaction
mixture was stirred overnight at room temperature in argon
atmosphere. After completion of reaction the reaction mixture was
diluted with mixed solvent (CHCl.sub.3:i-PrOH=4:1, 10 mL) and
washed with sat. NH.sub.4Cl. The organic layer was dried (anhydrous
sodium sulfate) and concentrated in vacuo. The crude product was
chromatographed on silica gel (Hexanes/EtOAc, 7:1) to yield target
compound 4. Yield 306 mg, 90%. R.sub.f=0.36, .sup.1H NMR
(CHCl.sub.3, 400 MHz): .delta. 7.91 (d, J=8.2, Hz, 2H), 7.38 (m,
3H), 7.15 (m, 4H), 3.91 (s, 3H), 3.80 (m, 4H), 3.50 (s, 2H), 0.85
(s, 9H), 0.02 (s, 6H); .sup.13C NMR (CHCl.sub.3, 100 MHz): .delta.
170.2, 167.0, 142.9, 140.8, 129.6, 129.5, 129.1, 128.6, 128.5,
128.0, 60.1, 52.1, 52.0, 41.4, 25.8, 18.2, -5.4; [M+H].sup.+=427.84
(APCI+).
4-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)(phenyl)amino)-2-oxoethyl)-N--
hydroxybenzamide (5)
[0247] Hydroxylamine (0.5 mL, 50% water solution) was added to a
solution containing methyl
4-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)(phenyl)amino)-2-oxoethyl)be-
nzoate 4 (60 mg, 0.140 mmol) in THF/MeOH (1:1, 1 mL). Reaction
mixture was treated with cat. amount of KCN (.about.0.5 mg) and
stirred at room temperature in argon atmosphere for 16 h. Then
solution was acidified by NH.sub.4Cl/HCl solution to pH 4. The
mixture was diluted with mixed solvent (CHCl.sub.3:i-PrOH=4:1, 10
mL) and washed with sat. NH.sub.4Cl. The organic layer was dried
(anhydrous sodium sulfate) and concentrated in vacuo. The crude
product was purified by preparative chromatography on silica gel
(CH.sub.2Cl.sub.2/MeOH, 10:1) to yield target compound 5. Yield 40
mg, 66%. R.sub.f=0.36, .sup.1H NMR (CD.sub.3OD, 400 MHz): .delta.
7.64 (d, J=8.4 Hz, 2H), 7.45 (m, 3H), 7.28 (d, J=8.0 Hz, 2H), 7.13
(d, J=8.0 Hz, 2H), 3.80 (m, 4H), 3.54 (s, 2H), 0.88 (s, 9H), 0.05
(s, 6H); .sup.13C NMR (CD.sub.3OD, 100 MHz): .delta. 172.0, 167.0,
142.6, 139.7, 130.8, 129.7, 128.8, 128.4, 127.6, 126.2, 60.7, 52.0,
41.0, 26.2, 18.0, -5.4; [M+H].sup.+=428.85 (APCI+).
N-hydroxy-4-(2-((2-hydroxyethyl)(phenyl)amino)-2-oxoethyl)
benzamide (6)
[0248] 4-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)
(phenyl)amino)-2-oxoethyl)-N-hydroxybenzamide 5 (16 mg, 0.037 mmol)
was dissolved in 5% TFA in CH.sub.2Cl.sub.2 (3 mL) and stirred for
5 min. Then the organic layer was concentrated in vacuo. The crude
product was purified by preparative chromatography on silica gel
(CH.sub.2Cl.sub.2/MeOH, 10:1) to yield target compound 5. Yield 8
mg, 68%. R.sub.f=0.23, .sup.1H NMR (CD.sub.3OD, 400 MHz): .delta.
7.64 (d, J=8.4 Hz, 2H), 7.45 (m, 3H), 7.29 (d, J=8.0 Hz, 2H), 7.14
(d, J=8.0 Hz, 2H), 3.86 (t, J=6.0 Hz, 2H), 3.68 (t, J=6.0 Hz, 2H),
3.55 (s, 2H); .sup.13C NMR (CD.sub.3OD, 100 MHz): .delta. 172.0,
166.9, 142.7, 139.7, 130.9, 129.9, 129.4, 128.7, 128.5, 127.1,
58.8, 51.8, 41.0; [M+H]'=315.26 (APCI+).
2-(4-(methoxycarbonyl)phenyl)acetic acid (8)
[0249] Methanol (6.38 g, 198.6 mmol), acetic acid (3.61 g, 60.18
mmol), calcium hypochlorite (4.30 g, 30.1 mmol) and molecular
sieves (8.6 g) were added sequentially to a solution containing
2-(4-(hydroxymethyl)phenyl)acetic acid (7, 1.00 g, 6.02 mmol) in
acetonitrile (50 mL). The reaction mixture was stirred in dark at
room temperature in argon atmosphere for 24 h. Another portion of
methanol (6.38 g, 198.6 mmol) and calcium hypochloride (1.72 g,
12.0 mmol) were added to the reaction mixture and stirred for 24 h.
NMR analysis showed complete consumption of both the starting
material and the intermediate aldehyde. Water (20 mL) and sodium
thiosulfate (4 g) were added to quench remaining hypochlorite.
Added 50 mL sat. NaHCO.sub.3 and filtered under suction. The
filterate was washed with ethylacetate (30 mL.times.2). The aqueous
layer was acidified with 2 N HCl to pH .about.2 and extracted with
ethylacetate (50 mL.times.3). Combined organic layer was dried
(anhydrous sodium sulfate) and concentrated in vacuo. The crude
product was chromatographed on silica gel (CH.sub.2Cl.sub.2/MeOH,
10:1) to yield target compound 8. Yield 740 mg, 63%. R.sub.f=0.53,
.sup.1H NMR (CHCl.sub.3, 400 MHz): .delta. 8.03 (d, J=8.3 Hz, 2H),
7.38 (d, J=8.3 Hz, 2H), 3.93 (s, 3H), 3.7 (s, 2H); .sup.13C NMR
(CHCl.sub.3, 100 MHz): .delta. 176.4, 166.8, 138.3, 129.9, 129.5,
129.3, 52.1, 40.8; [M+H].sup.+=195.03 (APCI+).
Example 2
Synthesis of Compound 14
[0250] HDAC inhibitor 14 was accessed according to the protocols
shown in Scheme 3. Alcohol 10 was prepared by treatment of amine 9
with glycolaldehyde followed by addition of sodium
triacetoxyborohydride. The alcohol 10 was then protected with
TBDMSCl followed by EDC coupling with acid 8 (Scheme 2). The methyl
ester 12 was converted directly, using a catalytic amount of KCN,
to the corresponding hydroxamic acid 13, which was deprotected with
TFA to afford compound 14.
##STR00034##
[0251] Reagents: a) 1. HOCH.sub.2CHO, DCE, 30 min. rt. 2.
NaBH(OAc).sub.3, 4 h. rt. b) TBDMS-Cl, DCM, 3 h. c) 8, EDC, DCM,
rt. 24 h. d) NH.sub.2OH, THF:MeOH (1:1), KCN (cat.) 16 h, rt. e) 5%
TFA in DCM, 5 min.
2-(quinolin-8-ylamino)ethanol (10)
[0252] Glycolaldehyde (437 mg, 7.29 mmol) was added to a solution
containing 8-aminoquinoline (9, 1.00 g, 6.94 mmol) in DCE (30 mL).
The solution was stirred at room temperature in argon atmosphere
for 30 min. Sodium triacetoxyborohydride (1.86 g, 3.84 mmol) was
then added portion-wise. The reaction mixture was further stirred
for 4 h and quenched with NaHCO.sub.3 to pH=9.0. The reaction
mixture was extracted with mixed solvent (CHCl.sub.3:i-PrOH=4:1, 30
mL.times.3). The organic layer was dried (sodium sulfate) and
concentrated in vacuo. The crude product was chromatographed on
silica gel (CH.sub.2Cl.sub.2/MeOH, 20:1) to yield target compound
10. Yield 0.950 g, 73%. R.sub.f=0.6, .sup.1H NMR (CHCl.sub.3, 400
MHz): .delta. 7.24 (dd, J=8.8, 7.4 Hz, 2H), 6.80 (t, J=7.4 Hz, 1H),
6.69 (d, J=8.8, 2H), 3.81 (t, J=5.2 Hz, 2H), 3.30 (t, J=5.2 Hz,
2H), 2.6 (br s, 1H); [M+H].sup.+=138.08 (APCI+)
N-(2-((tert-butyldimethylsilyl)oxy)ethyl)quinolin-8-amine (11)
[0253] TBDMS-Cl (624 mg, 4.13 mmol) and imidazole (648 mg, 9.51
mmol) was added to a solution containing
2-(quinolin-8-ylamino)ethanol 10 (600 mg, 3.19 mmol) in
CH.sub.2Cl.sub.2 (6 mL). The reaction mixture was stirred at room
temperature in argon atmosphere for 3 h. Then the reaction was
quenched with sat. NH.sub.4Cl. and washed with water (10
mL.times.2) and brine (10 mL.times.2). The organic layer was dried
(sodium sulfate) and concentrated in vacuo. The crude product was
chromatographed on silica gel (Hexanes/EtOAc, 7:1) to yield target
compound 11. Yield 730 g, 75%. R.sub.f=0.65, .sup.1H NMR
(CHCl.sub.3, 400 MHz): .delta. 8.73 (dd, J=4.2, 1.7 Hz, 1H), 8.07
(dd, J=8.2, 1.2 Hz, 1H), 7.39 (m, 2H), 7.07 (dd, J=8.2, 0.8 Hz,
1H), 6.73 (d, J=7.6, 1H), 6.50 (br s, 1H), 3.97 (t, J=5.9 Hz, 2H),
3.48 (t, J=5.9 Hz, 2H), 0.94 (s, 9H), 0.10 (s, 6H); .sup.13C NMR
(CHCl.sub.3, 100 MHz): .delta. 146.8, 144.9, 135.9, 128.7, 127.7,
121.3, 113.9, 104.8, 61.7, 45.5, 25.9, 18.3, -5.3;
[M+H].sup.+=303.25 (APCI+).
Methyl
4-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)(quinolin-8-yl)amino)--
2-oxoethyl)benzoate (12)
[0254] EDC (147 mg, 0.772 mmol) was added to a solution containing
N-(2-((tert-butyldimethylsilyl) oxy)ethyl)quinolin-8-amine 11 (156
mg, 0.545 mmol) and 2-(4-(methoxycarbonyl)phenyl)acetic acid 8,
(150 mg, 0.772 mmol) in CH.sub.2Cl.sub.2 (3 mL). The reaction
mixture was stirred overnight at room temperature in argon
atmosphere. After completion of reaction the reaction mixture was
diluted with mixed solvent (CHCl.sub.3:i-PrOH=4:1, 10 mL) and
washed with sat. NH.sub.4Cl. The organic layer was dried (sodium
sulfate) and concentrated in vacuo. The crude product was
chromatographed on silica gel (CH.sub.2Cl.sub.2/MeOH, 15:1) to
yield target compound 12. Yield 170 mg, 69%. R.sub.f=0.40, .sup.1H
NMR (CHCl.sub.3, 400 MHz): .delta. 8.93 (dd, J=4.1, 1.5 Hz, 1H),
8.28 (dd, J=8.3, 1.5 Hz, 1H), 7.87 (dd, J=8.2, 1.3 Hz, 1H), 7.82
(d, J=8.2, 2H), 7.64 (dd, J=7.3, 1.2 Hz, 1H), 7.51 (m, 2H), 7.02
(d, J=8.2, 2H), 4.39 (m, 1H), 3.96 (m, 1H), 3.73 (m, 1H), 3.55 (m,
1H), 3.35 (m, 2H), 1.8 (br s, 1H), 0.81 (s, 9H), 0.10 (s, 6H), 0.10
(s, 3H), 0.04 (s, 3H); .sup.13C NMR (CHCl.sub.3, 100 MHz): .delta.
171.2, 167.0, 151.0, 144.5, 141.1, 140.4, 136.2, 130.2, 129.4,
129.3, 129.1, 128.4, 128.1, 126.2, 121.8, 61.7, 52.3, 51.9, 41.6,
25.8, 18.1, -5.4, -5.5; [M+H].sup.+=479.29 (APCI+).
4-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)(quinolin-8-yl)amino)-2-oxoet-
hyl)-N-hydroxybenzamide (13)
[0255] Hydroxylanmine (0.5 mL, 50% water solution) was added to a
solution containing methyl methyl
4-(2-((2-((tert-butyldimethylsilyl) oxy)ethyl)
(quinolin-8-yl)amino)-2-oxoethyl)benzoate 12 (70 mg, 0.146 mmol) in
THF/MeOH (1:1, 1 mL). Reaction mixture was treated with cat. amount
of KCN (.about.0.5 mg) and stirred at room temperature in argon
atmosphere for 16 h. Then solution was acidified by NH.sub.4Cl/HCl
solution to pH 4. The mixture was diluted with diluted with mixed
solvent (CHCl.sub.3:i-PrOH=4:1, 10 mL) and washed with sat.
NH.sub.4Cl. The organic layer was dried (sodium sulfate) and
concentrated in vacuo. The crude product was purified by
preparative chromatography on silica gel (CH.sub.2Cl.sub.2/MeOH,
10:1) to yield target compound 13. Yield 45 mg, 64%. R.sub.f=0.60,
.sup.1H NMR (CHCl.sub.3, 400 MHz): .delta. 8.91 (d, J=3.3 Hz, 1H),
8.41 (d, J=8.1 Hz, 1H), 8.04 (d, J=8.1 Hz, 1H), 7.76 (d, J=7.2,
1H), 7.60 (m, 2H), 7.53 (d, J=8.2, 2H), 6.98 (d, J=7.9, 2H), 4.42
(m, 1H), 3.92 (m, 1H), 3.73 (m, 1H), 3.52 (m, 1H), 3.38 (s, 2H),
0.81 (s, 9H), 0.00 (s, 3H), -0.04 (s, 3H); .sup.13C NMR
(CHCl.sub.3, 100 MHz): .delta. 171.2, 167.0, 151.0, 144.5, 141.1,
140.4, 136.2, 130.2, 129.4, 129.3, 129.1, 128.4, 128.1, 126.2,
121.8, 61.7, 52.3, 51.9, 41.6, 18.1, -5.4, -5.5; [M+H].sup.+=480.46
(APCI+).
N-hydroxy-4-(2-((2-hydroxyethyl)(quinolin-8-yl)amino)-2-oxoethyl)
benzamide (14)
[0256] 4-(2-((2-((tert-butyldimethylsilyl)
oxy)ethyl)(quinolin-8-yl)amino)-2-oxoethyl)-N-hydroxybenzamide 13
(34 mg, 0.065 mmol) was dissolved in 5% TFA in CH.sub.2Cl.sub.2 (3
mL) and stirred for 5 min. Then the organic layer was concentrated
in vacuo. The crude product was purified by preparative
chromatography on silica gel (CH.sub.2Cl.sub.2/MeOH, 10:1) to yield
target compound 14. Yield 18 mg, 69%. R.sub.f=0.20, .sup.1H NMR
(CHCl.sub.3, 400 MHz): .delta. 8.89 (d, J=3.0 Hz, 1H), 8.50 (d,
J=8.2 Hz, 1H), 7.81 (d, J=6.7, 1H), 7.65 (m, 2H), 7.51 (d, J=8.0,
2H), 6.96 (d, J=8.0, 2H), 4.08 (m, 1H), 3.92 (m, 1H), 3.70 (m, 2H),
3.74 (s, 2H); .sup.13C NMR (CHCl.sub.3, 100 MHz): .delta. 171.2,
167.0, 150.1, 143.5, 139.1, 138.3, 137.7, 130.9, 130.2, 129.8,
129.3, 128.9, 126.8, 126.5, 122.1, 58.6, 51.7, 41.0;
[M+H].sup.+=366.33 (APCI+).
Example 3
Synthesis of Compounds with Various Linkers
[0257] HDAC inhibitor 19 is accessed according to the protocols
shown in Scheme 4. 4-Aminophenylacetic acid 15 is converted to
aniline 16 using methods similar to those described in Scheme 1.
Aniline 16 is coupled to acid 17 using EDC. The resulting methyl
carbamate derivative 18 is converted directly, using a catalytic
amount of KCN, to the corresponding hydroxamic acid, which is
deprotected with TFA to afford compound 19. Alternative coupling
partner, carboxylic acid 20, is also used to synthesize compound 21
(Scheme 5).
##STR00035##
##STR00036##
[0258] HDAC inhibitor 24 is accessed according to the protocols
shown in Scheme 6. 4-Aminophenylacetic acid 15 is converted to
aniline 16 using methods similar to those described in Scheme 1.
Aniline 16 is coupled to carbamic chloride derivative 22 under
basic conditions. The resulting urea-type derivative 23 is
converted to the corresponding N-hydroxy urea under hydrogenation
conditions, which is deprotected with TFA to afford compound 24.
Alternative coupling partner, monomethyl oxalyl chloride 25, is
also used to synthesize compound 26 (Scheme 7).
##STR00037##
##STR00038##
Example 4
Synthesis of Compound 26
[0259] HDAC inhibitor 26 was accessed according to the protocols
shown in Scheme 8. 4-Aminophenylacetic acid 15 was converted to
acid 27. Acid was coupled to substituted aniline 3. The resulting
amide derivative 28 was converted to HDAC inhibitor 26 using
methods previously described in Scheme 1.
##STR00039##
Example 5
Synthesis of Compound 24
[0260] HDAC inhibitor 24 was accessed according to the protocols
shown in Scheme 9. 4-Aminophenylacetic acid 15 was converted to
acid 30 using CDI in the presence of O-benzylhydroxylamine. Acid 30
was coupled to substituted aniline 3. The resulting amide
derivative 31 was converted to the N-hydroxy urea under
hydrogenation conditions, which was deprotected with TFA to afford
HDAC inhibitor 24.
##STR00040##
Example 6
Synthesis of Compound 21
[0261] HDAC inhibitor 21 was accessed according to the protocols
shown in Scheme 10. 4-Aminophenylacetic acid 15 was acylated with
the acid chloride of methylfumurate to form acid 32. Acid 32 was
coupled to substituted aniline 3. The resulting amide derivative 33
was converted to HDAC inhibitor 21 using methods previously
described in Scheme 1.
##STR00041##
Example 7
Synthesis of Compound 27
[0262] HDAC inhibitor 27 was accessed according to the protocols
shown in Scheme 11. Substituted aniline 3 was treated with
chloroacetyl chloride to form amide 35, which was substituted with
methyl 4-aminobenzoate to form compound 36. Methyl ester derivative
36 was converted to HDAC inhibitor 27 using methods previously
described in Scheme 1.
##STR00042##
Example 8
Synthesis of Compounds 28 and 29
[0263] HDAC inhibitors 28 and 29 were accessed according, to the
protocols shown in Scheme 12. Hydroxylamine 38 was N-acylated with
the corresponding acid chloride to form N-hydroxyamide 39, which
was coupled to aniline 3 followed by deprotection to form HDAC
inhibitors 28 and 29.
##STR00043##
Example 9
Synthesis of Compounds 30 and 32
[0264] HDAC inhibitor 30 was accessed according to the protocols
shown in Scheme 13. Methyl 4-aminobenzoate was acylated with with
chloroacetyl chloride to form amide 40, which was coupled with
aniline 3. The methyl ester derivative 41 was converted to HDAC
inhibitor 30 using methods previously described in Scheme 1. Methyl
4-hydroxybenzoate was converted to HDAC inhibitor 32 using a
similar sequence (Scheme 14).
##STR00044##
##STR00045##
Example 10
Synthesis of Compound 31
[0265] HDAC inhibitor 31 was accessed according to the protocols
shown in Scheme 15. 4-Aminophenylacetic acid 15 was acylated with
methyl malonyl chloride to form acid 43. Acid 43 was coupled to
substituted aniline 3. The resulting amide derivative 44 was
converted to HDAC inhibitor 31 using methods previously described
in Scheme 1.
##STR00046##
Example 11
Synthesis of Compound 33
[0266] HDAC inhibitor 33 was accessed according to the protocols
shown in Scheme 16. 4-(Methoxycarbonyl)benzoic acid was coupled to
substituted aniline 3. The resulting amide derivative 49 was
converted to HDAC inhibitor 33 using methods previously described
in Scheme 1.
##STR00047##
Example 12
Synthesis of Compounds 34, 35 and 36
[0267] HDAC inhibitor 34 was accessed according to the protocols
shown in Scheme 17. 4-(2-Methoxy-2-oxoethyl)benzoic acid was
coupled to substituted aniline 3. The resulting amide derivative 51
was converted to HDAC inhibitor 34 using methods previously
described in Scheme 1. 4-(3-Methoxy-3-oxopropyl)benzoic acid and
2-(4-(3-methoxy-3-oxopropyl)phenyl) acetic acid were converted to
HDAC inhibitors 35 and 36, respectively, using a similar sequence
(Schemes 18 and 19).
##STR00048##
##STR00049##
##STR00050##
Example 13
Ar.sub.1 Derivatives
[0268] HDAC inhibitors with various Ar.sub.1 groups were
synthesized according to the protocols shown in Schemes 20-25.
Anline 3 was acylated with chloroacetyl chloride to form alkyl
chloride 35. Chloride 35 was substituted with the corresponding
aniline-type derivative to form methyl esters 59, 62, 64, 66, 68
and 70, which were then converted to the HDAC inhibitors 37, 38,
39, 40, 41, 42 and 43 using methods previously described in Scheme
1
##STR00051##
##STR00052##
##STR00053##
##STR00054##
##STR00055##
##STR00056##
Example 14
Synthesis of Compound 79
[0269] HDAC inhibitor 79 was accessed according to the protocols
shown in Scheme 25. 2-(3-(methoxycarbonyl)phenyl)acetic acid 77 was
coupled to substituted aniline 3. The resulting amide derivative 78
was converted to HDAC inhibitor 79 using methods previously
described in Scheme 1 (Scheme 25).
##STR00057##
Methyl 2-(3-bromophenyl)acetate (73)
[0270] 2-(3-bromophenyl)acetic acid (1.00 g, 4.65 mmol) was
dissolved in freshly distilled methanol (50 mL). Added conc.
H.sub.2SO.sub.4 (8 drops) and refluxed in a Dean-Stark apparatus
for 3 hrs. Reaction mixture was cooled to room temperature and
concentrated in vacuo. The crude product was dissolved in
CH.sub.2Cl.sub.2 (40 mL) and washed with sat. NaHCO.sub.3 (30
mL.times.2) and sat. NaCl (30 mL). The organic layer was dried
(anhydrous sodium sulfate) and concentrated in vacuo. The crude
product was chromatographed on silica gel (Hexanes/EtOAc, 5:1) to
yield target compound 73. Yield 1.011 g, 95%. R.sub.f=0.60, .sup.1H
NMR (400 MHz, Chloroform-d) .delta. 7.47-7.43 (m, 1H), 7.41 (dt,
J=6.5, 2.2 Hz, 1H), 7.24-7.16 (m, 2H), 3.70 (s, 3H), 3.60 (s, 2H);
[M+H]+=230.12 (APCI+).
Methyl 2-(3-cyanophenyl)acetate (74)
[0271] A mixture of 2-(3-bromophenyl)acetic acid 73, (1.0 g, 4.36
mmol) and CuCN (0.43 g, 4.80 mmol) in anhydrous DMF (10 mL) was
refluxed overnight. The mixture was cooled to room temperature and
poured into ice-water (100 mL), and the resulting precipitate was
filtered. The solid was transferred to a beaker containing
ethylenediamine (25 mL) and H.sub.2O (10 mL), and the precipitate
formed was extracted with CH.sub.2Cl.sub.2 (30 mL). The organic
layer was washed with sat. NaCl (30 mL.times.2), dried (anhydrous
sodium sulfate) and concentrated in vacuo. The crude product was
chromatographed on silica gel (Hexanes/EtOAc, 5:1) to yield target
compound 74. Yield 580 mg, 75%. R.sub.f=0.38, .sup.1H NMR (400 MHz,
Methanol-d4) .delta. 7.69 (d, J=1.7 Hz, 1H), 7.64 (ddt, J=8.5, 7.0,
1.4 Hz, 2H), 7.52 (t, J=7.7 Hz, 1H), 3.78 (s, 3H), 3.73 (s, 2H).;
[M+H]+=176.13 (APCI+).
3-(carboxymethyl)benzoic acid (75)
[0272] 6M Sodium hydroxide (10 mL) was added to methyl
2-(3-cyanophenyl)acetate 74, (550 mg, 3.14 mmol) in methanol (10
mL) and then heated at 90.degree. C. overnight. After concentrating
the reaction mixture, the aqueous layer was washed with
CH.sub.2Cl.sub.2 (20 mL.times.2), then acidified to pH-3 with 12M
HCl. The precipitate was extracted with ethyl acetate and washed
with sat. NaCl (30 mL.times.2), dried (anhydrous sodium sulfate)
and concentrated in vacuo. The crude product was chromatographed on
silica gel (CH.sub.2Cl.sub.2/MeOH, 20:1) to yield target compound
75. Yield 508 mg, 90%. R.sub.f=0.30, .sup.1H NMR (400 MHz,
Methanol-d.sub.4) .delta. 8.06-7.87 (m, 2H), 7.54 (tt, J=5.7, 1.4
Hz, 1H), 7.45 (t, J=7.6 Hz, 1H), 3.71 (s, 2H); [M+H]+=181.15
(APCI+).
Methyl 3-(2-methoxy-2-oxoethyl)benzoate (76)
[0273] 3-(carboxymethyl) benzoicacid (0.500 g, 2.77 mmol) was
dissolved in freshly distilled methanol (25 mL). Added conc.
H.sub.2SO.sub.4 (8 drops) and refluxed in a Dean-Stark apparatus
for 16 hrs. Reaction mixture was cooled to room temperature and
concentrated in vacuo. The crude product was dissolved in
CH.sub.2Cl.sub.2 (40 mL) and washed with sat. NaHCO.sub.3 (30
mL.times.2) and sat. NaCl (30 mL). The organic layer was dried
(anhydrous sodium sulfate) and concentrated in vacuo. The crude
product was chromatographed on silica gel (Hexanes/EtOAc, 5:1) to
yield target compound 76. Yield 547 mg, 95%. R.sub.f=0.60, .sup.1H
NMR (400 MHz, Chloroform-d) .delta. 7.98 (dd, J=6.2, 1.6 Hz, 2H),
7.51 (dt, J=7.6, 1.6 Hz, 1H), 7.47-7.40 (m, 1H), 3.94 (s, 3H), 3.73
(s, 3H), 3.71 (s, 2H); [M+H]+=209.25 (APCI+).
2-(3-(methoxycarbonyl)phenyl)acetic acid (77)
[0274] A solution of K.sub.2CO.sub.3 (700 mg, 5.07 mmol) in
H.sub.2O (10 mL) was added to a solution of methyl
3-(2-methoxy-2-oxoethyl)benzoate 76, (500 mg, 2.40 mmol) in
methanol (10 mL) and stirred overnight at room temperature.
Reaction mixture was concentrated and dissolved in water (20 mL).
Aqueous later was extracted with EtOAc to the unreacted diester 76,
acidified with 1N HCl, and extracted with EtOAc (30 mL.times.2).
Organic layer was dried (anhydrous sodium sulfate) and concentrated
in vacuo. The crude product was chromatographed on silica gel
(Hexanes/EtOAc, 5:1) to yield target compound 77. Yield 508 mg,
90%. R.sub.f=0.36, .sup.1H 1H NMR (400 MHz, Methanol-d.sub.4)
.delta. 7.97 (d, J=1.7 Hz, 1H), 7.91 (dt, J=7.8, 1.4 Hz, 1H),
7.57-7.49 (m, 1H), 7.43 (t, J=7.6 Hz, 1H), 3.91 (s, 3H), 3.67 (s,
2H); [M+H]+=195.14 (APCI+).
Methyl 3-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)(phenyl)
amino)-2-oxoethyl)benzoate (78)
[0275] EDC (511 mg, 2.67 mmol) was added to a solution containing
N-(2-((tertbutyldimethylsilyl)oxy)ethyl)aniline 3, (535 mg, 2.13
mmol) and 2-(3-(methoxycarbonyl)phenyl)acetic acid 77, (345 mg,
1.78 mmol) in CH.sub.2Cl.sub.2 (10 mL). The reaction mixture was
stirred overnight at room temperature in argon atmosphere. After
completion of reaction the reaction mixture was diluted with mixed
solvent (CHCl.sub.3:i-PrOH=4:1, 10 mL) and washed with sat.
NH.sub.4Cl. The organic layer was dried (anhydrous sodium sulfate)
and concentrated in vacuo. The crude product was chromatographed on
silica gel (Hexanes/EtOAc, 7:1) to yield target compound 78. Yield
732 mg, 70%. R.sub.f=0.35, .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 7.90 (ddd, J=5.6, 3.4, 1.7 Hz, 1H), 7.71 (s, 1H), 7.48-7.31
(m, 6H), 7.23-7.19 (m, 2H), 3.92 (s, 3H), 3.87-3.76 (m, 4H), 0.86
(s, 9H), 0.03 (s, 6H); [M+H]+=427.61 (APCI+).
3-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)(phenyl)amino)-2-oxoethyl)-N--
hydroxybenzamide
[0276] Hydroxylamine (1 mL, 50% water solution) was added to a
solution containing methyl
3-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)
(phenyl)amino)-2-oxoethyl) benzoate (200 mg, 0.468 mmol) in
THF/MeOH (1:1, 2 mL). Reaction mixture was treated with cat. amount
of KCN (.about.0.5 mg) and stirred at room temperature in argon
atmosphere for 16 h. Then solution was acidified by NH.sub.4Cl/HCl
solution to pH 4. The mixture was diluted with mixed solvent
(CHCl.sub.3:i-PrOH=4:1, 10 mL) and washed with sat. NH.sub.4Cl. The
organic layer was dried (anhydrous sodium sulfate) and concentrated
in vacuo. The crude product was purified by preparative
chromatography on silica gel (CH.sub.2Cl.sub.2/MeOH, 10:1) to yield
3-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)
(phenyl)amino)-2-oxoethyl)-N hydroxylbenzamide. Yield 40 mg, 66%.
R.sub.f=0.36, .sup.1H NMR (400 MHz, Methanol-d.sub.4) .delta. 7.60
(dt, J=7.7, 1.5 Hz, 1H), 7.53-7.38 (m, 4H), 7.39-7.26 (m, 3H),
7.25-7.18 (m, 1H), 3.85 (dd, J=6.8, 5.3 Hz, 2H), 3.77 (dd, J=6.1,
4.7 Hz, 2H), 3.53 (s, 2H), 0.87 (s, 8H), 0.04 (s, 6H);
[M+H]+=429.28 (APCI+).
N-hydroxy-3-(2-((2-hydroxyethyl)(phenyl)amino)-2-oxoethyl)
benzamide (79)
[0277] 3-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)
(phenyl)amino)-2-oxoethyl)-N-hydroxybenzamide,
3-(2-((2-((tert-butyldimethylsilyl)oxy)ethyl)
(phenyl)amino)-2-oxoethyl)-N-hydroxybenzamide (60 mg, 0.14 mmol)
was dissolved in 2% HCl in EtOH (5 mL) and stirred for 3 h. Then
the reaction mixture was concentrated in vacuo. The crude product
was purified by preparative chromatography on silica gel
(CH.sub.2Cl.sub.2/MeOH, 10:1) to yield target compound 79. Yield 24
mg, 54%. R.sub.f=0.28, .sup.1H NMR (400 MHz, Methanol-d.sub.4)
.delta. 7.59 (d, J=7.7 Hz, 1H), 7.53-7.39 (m, 4H), 7.39-7.26 (m,
3H), 7.21 (d, J=7.7 Hz, 1H), 3.86 (t, J=6.0 Hz, 2H), 3.68 (t, J=6.0
Hz, 2H), 3.53 (s, 2H).; .sup.13C NMR (101 MHz, Methanol-d.sub.4)
.delta. 171.80, 166.64, 142.24, 135.94, 132.21, 129.52, 128.33,
128.24, 128.18, 127.57, 125.03, 58.38, 51.46, 40.52.; [M+H]+
Calc.=315.1345. Found=315.1336 (FAB+).
Example 15
Biological Data
[0278] Recombinant HDAC1 and HDAC6 are used to evaluate the
selective inhibitory potency of each compound. Cell based assays:
Normal cells (Human Foreskin Fibroblast cells), LNCaP (human
prostate cancer cells), MCF-7 (human breast cancer cells, A549
(human adenocarcinoma of lung cells) and ARP-1 (human multiple
myeloma cells) are used in these assays. Cells were cultured for up
to 72 hr without and with the potential HDAC6-selective inhibitor.
SAHA was used as a control. Cell number and cell viability were
determined by enumeration. Proteins were extracted from cells and
assayed for accumulation of acetylated tubulin and acetylated
histones. All methods are described in Namdar et al., PNAS, 2010,
107:20003-8. In vivo animal studies: Potential HDAC6 inhibitor
compounds are further assayed by administration to mice for up to 5
days with daily injections. Animals are sacrificed and tissues are
analyzed for accumulation of acetylated tubulin and acetylated
histones.
DISCUSSION
[0279] HDAC6-selective inhibitors were identified on the basis of
accumulation of acetylated tubulin without accumulation of
acetylated histones. Compound 6 and 14 were identified as
HDAC6-selective inhibitors. As show in FIG. 2, significant levels
of tubulin acylation were present without induction of H3
acetylation. The levels of tubulin acetylation that can be reached
with compound 6 in the absence of histone acetylation were greater
than selective levels with tubacin.
[0280] As indicated by the cell based (LNCaP-human prostate)
assays, compounds 6 inhibits cell number (FIG. 3) but does not
decrease cell viability (FIG. 4). Unlike SAHA, which kills LNCaP
cells, there is no detectable death of LNCaP cells with compound 6
even at concentrations as high as 32 .mu.M.
[0281] An additional aspect of the invention provides synthetic
methods and chemical intermediates that may be used to synthesize
additional HDAC inhibitors. Additional compounds, which are
synthesized according to Schemes 4-7 or according to methods known
in the art, are expected to function analogously to compound 6.
Compounds with a variety of "Z" linkers are expected to function
analogously to compound 6. The left hand portion of the compound
(R.sub.1, R.sub.2 and R.sub.3) is also varied in order to improve
solubility and drug-like properties and is expected to function
analogously to compound 6.
[0282] The compounds of the present invention are HDAC6 selective
inhibitors. Compounds 6, 14 and 79 were HDAC inhibitors selective
for HDAC6 relative to HDAC1. Compounds 21, 24, 26, 27, 28, 29, 30,
31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, and 43 are HDAC
inhibitors selective for HDAC6 relative to HDAC1.
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* * * * *