U.S. patent application number 15/203023 was filed with the patent office on 2017-04-06 for pyrimidine hydroxy amide compounds as protein deacetylase inhibitors and methods of use thereof.
The applicant listed for this patent is ACETYLON PHARMACEUTICALS, INC.. Invention is credited to Yun Cao, Yanbing Ding, Yong Liu, Ralph Mazitschek, John H. van Duzer, Nan Yu.
Application Number | 20170096403 15/203023 |
Document ID | / |
Family ID | 46047943 |
Filed Date | 2017-04-06 |
United States Patent
Application |
20170096403 |
Kind Code |
A1 |
van Duzer; John H. ; et
al. |
April 6, 2017 |
PYRIMIDINE HYDROXY AMIDE COMPOUNDS AS PROTEIN DEACETYLASE
INHIBITORS AND METHODS OF USE THEREOF
Abstract
The present invention relates to novel pyrimidine hydroxy amide
compounds, and the use of such compounds in the inhibition of HDAC6
and in the treatment of various diseases, disorders or conditions
related to HDAC6.
Inventors: |
van Duzer; John H.;
(Georgetown, MA) ; Mazitschek; Ralph; (Belmont,
MA) ; Ding; Yanbing; (Richmond, CA) ; Yu;
Nan; (Pudong, CN) ; Cao; Yun; (Pudong, CN)
; Liu; Yong; (Pudong, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
ACETYLON PHARMACEUTICALS, INC. |
Boston |
MA |
US |
|
|
Family ID: |
46047943 |
Appl. No.: |
15/203023 |
Filed: |
July 6, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14082472 |
Nov 18, 2013 |
9409890 |
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15203023 |
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13296748 |
Nov 15, 2011 |
8614223 |
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14082472 |
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61503286 |
Jun 30, 2011 |
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61414158 |
Nov 16, 2010 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 7/06 20180101; A61P
21/00 20180101; A61P 25/32 20180101; C07D 403/12 20130101; A61P
21/02 20180101; C07D 405/12 20130101; C07D 409/12 20130101; C07D
401/14 20130101; A61P 35/02 20180101; A61P 17/06 20180101; A61P
11/00 20180101; A61P 25/00 20180101; A61P 17/04 20180101; A61P
35/00 20180101; A61P 31/04 20180101; A61P 19/02 20180101; A61P
19/08 20180101; A61P 37/08 20180101; A61P 1/16 20180101; A61P 27/02
20180101; A61P 29/00 20180101; A61P 13/12 20180101; A61P 43/00
20180101; A61P 1/02 20180101; A61P 11/02 20180101; A61P 9/00
20180101; A61P 3/10 20180101; A61P 27/14 20180101; A61P 19/06
20180101; A61P 39/04 20180101; C07D 239/42 20130101; A61P 11/06
20180101; A61P 17/00 20180101; A61P 25/16 20180101; A61P 1/04
20180101; A61P 21/04 20180101; C07D 401/12 20130101; A61P 9/10
20180101; A61P 17/02 20180101; A61P 37/02 20180101; A61P 7/00
20180101; A61P 5/14 20180101; A61P 27/16 20180101; A61P 25/28
20180101; A61P 25/14 20180101; A61P 37/06 20180101 |
International
Class: |
C07D 239/42 20060101
C07D239/42; C07D 401/14 20060101 C07D401/14; C07D 409/12 20060101
C07D409/12; C07D 401/12 20060101 C07D401/12; C07D 405/12 20060101
C07D405/12 |
Claims
1. A compound of formula I: ##STR00190## or a pharmaceutically
acceptable salt, ester or prodrug thereof, wherein, the ring
comprising X.sub.1, X.sub.2, X.sub.3, and X.sub.4 is selected from
the following: ##STR00191## R.sub.1 is H, optionally substituted
alkyl, optionally substituted aryl, optionally substituted
arylalkyl optionally substituted heteroarylalkyl, or optionally
substituted heteroaryl; and R.sub.2 is H or alkyl; wherein R.sub.1
and R.sub.2 are both not simultaneously H.
2-4. (canceled)
5. The compound of claim 4, wherein R.sub.1 is H, methyl, ethyl,
propyl, i-propyl, butyl, i-butyl, t-butyl, pentyl, hexyl, phenyl,
benzyl, phenethyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, piperidinyl, pyrrolidinyl, tetrahydropyranyl,
thiophenyl, or tetrahydrofuranyl, each of which may be optionally
substituted.
6. The compound of claim 5, wherein R.sub.1 is substituted with 1-4
groups independently selected from optionally substituted alkyl,
optionally substituted aryl, optionally substituted heteroaryl,
optionally substituted heterocycloalkyl, halo, haloalkyl, alkoxy,
haloalkoxy, and nitro.
7. The compound of claim 6, wherein R.sub.1 is substituted with 1-4
groups independently selected from methyl, phenyl, pyridyl,
morpholino, indolyl, pyrazinyl, F, Cl, Br, methoxy, OCF.sub.3, and
trifluoromethyl; each of which may be further substituted by alkyl,
aryl, haloalkyl, haloalkoxy, halo, or alkoxy.
8-16. (canceled)
17. The compound of claim 1, of formula III: ##STR00192## or a
pharmaceutically acceptable salt, ester or prodrug thereof,
wherein, R.sub.2 is H or methyl; R.sub.1 is H, methyl, ethyl,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, piperidinyl,
pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl, benzyl,
phenethyl, pyridyl, or phenyl, each of which may be optionally
substituted with 1-4 groups independently selected from methyl,
phenyl, pyridyl, morpholino, indolyl, pyrazinyl, F, Cl, Br,
methoxy, OCF.sub.3, trifluoromethyl, OH, NH.sub.2, CN, and
NO.sub.2, each of which may be further substituted by alkyl, aryl,
haloalkyl, halo, or alkoxy.
18-21. (canceled)
22. The compound of claim 17, wherein R.sub.1 is phenyl or
pyridyl.
23. (canceled)
24. The compound of claim 22, wherein R.sub.1 is further
substituted with 1-4 substituents independently selected from
methyl, F, Cl, OCH.sub.3, CF.sub.3, OCF.sub.3, phenyl, pyridyl,
morpholinyl, or NO.sub.2.
25. The compound of claim 1, of formula IV: ##STR00193## or a
pharmaceutically acceptable salt, ester or prodrug thereof,
wherein, R.sub.2 is H or methyl; R.sub.x is H, alkyl, haloalkyl,
alkenyl, aryl, arylalkyl, heteroarylalkyl, heteroaryl,
heterocyclic, carbocyclic, --C(O)R.sub.5, or --S(O).sub.pR.sub.5;
each of which may be optionally substituted; R.sub.y is H, alkyl,
haloalkyl, alkenyl, aryl, arylalkyl, heteroarylalkyl, heteroaryl,
heterocyclic, carbocyclic, --C(O)R.sub.5, or --S(O).sub.pR.sub.5;
each of which may be optionally substituted; or R.sub.x and R.sub.y
together with the carbon to which each is attached, forms a
cycloalkyl or heterocycloalkyl, each of which is optionally
substituted; each R.sub.A is independently alkyl, alkoxy,
cycloalkyl, aryl, heterocycloalkyl, heteroaryl, arylalkyl,
heteroarylalkyl, haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN,
or --NH.sub.2; or two R.sub.A groups together with the atoms to
which each is attached, can form an optionally substituted ring;
each R.sub.5 is independently H, alkyl, carbocycle, heterocycle,
aryl, or optionally substituted heteroaryl; m is 0, 1, 2, 3, or 4;
and p is 0, 1, 2, or 3.
26-37. (canceled)
38. The compound of claim 1, of formula VI: ##STR00194## or a
pharmaceutically acceptable salt, ester or prodrug thereof,
wherein, R.sub.x is H, alkyl, or aryl; each of which is optionally
substituted; R.sub.y is H or alkyl; each of which is optionally
substituted; each R.sub.A is independently alkyl, alkoxy,
cycloalkyl, aryl, heterocycloalkyl, heteroaryl, arylalkyl,
heteroarylalkyl, haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN,
or --NH.sub.2; and m is 0, 1, or 2.
39. The compound of claim 38, wherein R.sub.x is H, methyl, ethyl,
or phenyl.
40. The compound of claim 38, wherein R.sub.y is H, methyl or
ethyl.
41. The compound of claim 38, wherein m is 1 or 2, and each R.sub.A
is independently methyl, phenyl, F, Cl, methoxyl, or CF.sub.3.
42. (canceled)
43. The compound of claim 1, of formula VII-A: ##STR00195## or a
pharmaceutically acceptable salt, ester or prodrug thereof,
wherein, ring A is a heteroaryl or heterocyclic; R.sub.2 is H or
methyl; R.sub.x is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; R.sub.y is H, alkyl, haloalkyl, alkenyl,
aryl, arylalkyl, heteroarylalkyl, heteroaryl, heterocyclic,
carbocyclic, --C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which
may be optionally substituted; or R.sub.x and R.sub.y together with
the carbon to which each is attached, form a cycloalkyl or
heterocycloalkyl, each of which is optionally substituted; each
R.sub.A is independently alkyl, alkoxy, cycloalkyl, aryl,
heterocycloalkyl, heteroaryl, arylalkyl, heteroarylalkyl,
haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN, or --NH.sub.2;
each R.sub.5 is independently H, alkyl, carbocycle, heterocycle,
aryl, or optionally substituted heteroaryl; and m is 0, 1, 2, 3, or
4.
44. The compound of claim 43, wherein ring A is acridinyl,
azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl,
benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl,
benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl,
carbazolyl, NH-carbazolyl, carbolinyl, chromanyl, chromenyl,
cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,
dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl,
imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl,
indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl,
isochromanyl, isoindazolyl, isoindolinyl, isoindolyl,
isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl,
naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl,
1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl; 1,2,5-oxadiazolyl,
1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl,
pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl,
phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl,
piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl,
pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole,
pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl,
pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl,
quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,
tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,
6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl,
1,2,5-thiadiazolyl, 1,3,4thiadiazolyl, thianthrenyl, thiazolyl,
thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl,
thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl,
1,2,5-triazolyl, 1,3,4-triazolyl, or xanthenyl.
45. (canceled)
46. The compound of claim 43, wherein R.sub.x is H, alkyl, aryl, or
heteroaryl.
47. The compound of claim 43, wherein R.sub.y is H, alkyl, aryl, or
heteroaryl.
48. The compound of claim 43, wherein R.sub.x is H, methyl or
pyridine, and R.sub.y is H.
49. The compound of claim 43, wherein R.sub.x and R.sub.y together
with the carbon to which each is attached, forms a cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl,
each of which is optionally substituted.
50. The compound of claim 49, wherein R.sub.x and R.sub.y together
with the carbon to which each is attached, forms tetrahydropyran,
piperidine, piperazine, morpholine, tetrahydrofuran,
tetrahydrothiophene, pyrrolidine, oxozolidine, or imidazolidine,
each of which is optionally substituted.
51. The compound of claim 43, wherein R.sub.A is H, alkyl, alkoxy,
cycloalkyl, aryl, heterocycloalkyl, heteroaryl, arylalkyl,
heteroarylalkyl, haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN,
or --NH.sub.2; and m is 0 or 1.
52-56. (canceled)
57. The compound of claim 1, selected from the following:
##STR00196## ##STR00197## ##STR00198## ##STR00199## ##STR00200##
##STR00201## ##STR00202## ##STR00203## ##STR00204## ##STR00205##
##STR00206## ##STR00207## ##STR00208## ##STR00209## ##STR00210##
##STR00211## ##STR00212## ##STR00213## ##STR00214## ##STR00215##
##STR00216## ##STR00217## ##STR00218## ##STR00219##
58. (canceled)
59. A pharmaceutical composition comprising a compound of claim 1,
or a pharmaceutically acceptable ester, salt, or prodrug thereof,
together with a pharmaceutically acceptable carrier.
60. A method of selectively inhibiting HDAC6 over other HDACs in a
subject, comprising administering to the subject a compound of
claim 1 or a pharmaceutically acceptable salt, ester or prodrug
thereof.
61-63. (canceled)
64. A method of treating a disease mediated by HDAC6 in a subject
comprising administering to the subject a compound of claim 1 or a
pharmaceutically acceptable salt, ester or prodrug thereof.
65-74. (canceled)
Description
RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. provisional
application 61/414,158, filed on Nov. 16, 2010, and U.S.
provisional application 61/503,286, filed on Jun. 30, 2011. The
entire contents of the aforementioned applications are incorporated
herein by reference.
BACKGROUND OF THE INVENTION
[0002] The identification of small organic molecules that affect
specific biological functions is an endeavor that impacts both
biology and medicine. Such molecules are useful as therapeutic
agents and as probes of biological function. Such small molecules
have been useful at elucidating signal transduction pathways by
acting as chemical protein knockouts, thereby causing a loss of
protein function. (Schreiber et al, J. Am. Chem. Soc, 1990, 112,
5583; Mitchison, Chem. and Biol., 1994, I5 3) Additionally, due to
the interaction of these small molecules with particular biological
targets and their ability to affect specific biological function
(e.g. gene transcription), they may also serve as candidates for
the development of new therapeutics.
[0003] One biological target of recent interest is histone
deacetylase (HDAC) (see, for example, a discussion of the use of
inhibitors of histone deacetylases for the treatment of cancer:
Marks et al. Nature Reviews Cancer 2001, 7,194; Johnstone et al.
Nature Reviews Drug Discovery 2002, 287). Post-translational
modification of proteins through acetylation and deacetylation of
lysine residues plays a critical role in regulating their cellular
functions. HDACs are zinc hydrolases that modulate gene expression
through deacetylation of the N-acetyl-lysine residues of histone
proteins and other transcriptional regulators (Hassig et al Curr.
Opin. Chem. Biol. 1997, 1, 300-308). HDACs participate in cellular
pathways that control cell shape and differentiation, and an HDAC
inhibitor has been shown effective in treating an otherwise
recalcitrant cancer (Warrell et al J. Natl. Cancer Inst. 1998, 90,
1621-1625). At this time, eleven human HDACs, which use Zn as a
cofactor, have been identified (Taunton et al. Science 1996, 272,
408-411; Yang et al. J. Biol. Chem. 1997, 272, 28001-28007.
Grozinger et al. Proc. Natl. Acad. Sci. U.S.A. 1999, 96, 4868-4873;
Kao et al. Genes Dev. 2000, 14, 55-66. Hu et al J. Biol. Chem.
2000, 275, 15254-15264; Zhou et al. Proc. Natl. Acad. Sci. U.S.A.
2001, 98, 10572-10577; Venter et al. Science 2001, 291, 1304-1351)
these members fall into three classes (class I, II, and IV). An
additional seven HDACs have been identified which use NAD as a
cofactor. To date, no small molecules are known that selectively
target any particular class or individual members of this family
(for example ortholog-selective HDAC inhibitors have been reported:
(a) Meinke et al. J. Med. Chem. 2000, 14, 4919-4922; (b) Meinke, et
al Curr. Med. Chem. 2001, 8, 211-235). There remains a need for
preparing structurally diverse HDAC and tubulin deacetylase (TDAC)
inhibitors particularly ones that are potent and/or selective
inhibitors of particular classes of HDACs or TDACs and individual
HDACs and TDACs.
[0004] Recently, a cytoplasmic histone deacetylase protein, HDAC6,
was identified as necessary for aggresome formation and for
survival of cells following ubiquitinated misfolded protein stress.
The aggresome is an integral component of survival in cancer cells.
The mechanism of HDAC6-mediated aggresome formation is a
consequence of the catalytic activity of the carboxy-terminal
deacetylase domain, targeting an uncharacterized non-histone
target. The present invention also provides small molecule
inhibitors of HDAC6. In certain embodiments, these new compounds
are potent and selective inhibitors of HDAC6.
[0005] The aggresome was first described in 1998, when it was
reported that there was an appearance of microtubule-associated
perinuclear inclusion bodies in cells over-expressing the
pathologic .DELTA.F508 allele of the cystic fibrosis transmembrane
conductance receptor (CFTR). Subsequent reports identified a
pathologic appearance of the aggresome with over-expressed
presenilin-1 (Johnston J A, et al. J Cell Biol. 1998;
143:1883-1898), parkin (Junn E, et al. J Biol Chem. 2002; 277:
47870-47877), peripheral myelin protein PMP22 (Notterpek L, et al.
Neurobiol Dis. 1999; 6: 450-460), influenza virus nucleoprotein
(Anton L C, et al. J Cell Biol. 1999; 146:113-124), a chimera of
GFP and the membrane transport protein pi 15 (Garcia-Mata R, et al.
J Cell Biol. 1999; 146: 1239-1254) and notably amyloidogenic light
chains (Dui J L, et al. J Cell Biol. 2001; 152:705-716). Model
systems have been established to study ubiquitinated (.DELTA.F508
CFTR) (Johnston J A, et al. J Cell Biol. 1998; 143:1883-1898) and
non-ubiquitinated (GFP-250) (Garcia-Mata R, et al. J Cell Biol.
1999; 146:1239-1254) protein aggregate transport to the aggresome.
Secretory, mutated, and wild-type proteins may assume unstable
kinetic intermediates resulting in stable aggregates incapable of
degradation through the narrow channel of the 26S proteasome. These
complexes undergo active, retrograde transport by dynein to the
pericentriolar aggresome, mediated in part by a cytoplasmic histone
deacetylase, HDAC6 (Kawaguchi Y, et al. Cell. 2003;
115:727-738).
[0006] Histone deacetylases are a family of at least 11
zinc-binding hydrolases, which catalyze the deacetylation of lysine
residues on histone proteins. HDAC inhibition results in
hyperacetylation of chromatin, alterations in transcription, growth
arrest, and apoptosis in cancer cell lines. Early phase clinical
trials with available nonselective HDAC inhibitors demonstrate
responses in hematologic malignancies including multiple myeloma,
although with significant toxicity. Of note, in vitro synergy of
conventional chemotherapy agents (such as melphalan) with
bortezomib has been reported in myeloma cell lines, though dual
proteasome-aggresome inhibition was not proposed. Until recently
selective HDAC inhibitors have not been realized.
[0007] HDAC6 is required for aggresome formation with ubiquitinated
protein stress and is essential for cellular viability in this
context. HDAC6 is believed to bind ubiquitinated proteins through a
zinc finger domain and interacts with the dynein motor complex
through another discrete binding motif. HDAC6 possesses two
catalytic deacetylase domains. It is not presently known whether
the amino-terminal histone deacetylase or the carboxy-terminal
tubulin deacetylase (TDAC) domain mediates aggresome formation.
[0008] Aberrant protein catabolism is a hallmark of cancer, and is
implicated in the stabilization of oncogenic proteins and the
degradation of tumor suppressors (Adams J. Nat Rev Cancer. 2004;
4:349-360). Tumor necrosis factor alpha induced activation of
nuclear factor kappa B (NFKB) is a relevant example, mediated by
NFKB inhibitor beta (IKB) proteolytic degradation in malignant
plasma cells. The inhibition of IKB catabolism by proteasome
inhibitors explains, in part, the apoptotic growth arrest of
treated myeloma cells (Hideshima T, et al. Cancer Res. 2001;
61:3071-3076). Multiple myeloma is an ideal system for studying the
mechanisms of protein degradation in cancer. Since William Russell
in 1890, cytoplasmic inclusions have been regarded as a defining
histological feature of malignant plasma cells. Though the precise
composition of Russell bodies is not known, they are regarded as
ER-derived vesicles containing aggregates of monotypic
immunoglobulins (Kopito R R, Sitia R. EMBO Rep. 2000: 1:225-231)
and stain positive for ubiquitin (Manetto V, et al. Am J Pathol.
1989; 134:505-513). Russell bodies have been described with CFTR
over-expression in yeast (Sullivan M L, et al. J. Histochem.
Cytochem. 2003; 51:545-548), thus raising the suspicion that these
structures may be linked to overwhelmed protein catabolism, and
potentially the aggresome. The role of the aggresome in cancer
remains undefined.
[0009] Aberrant histone deacetylase activity has also been linked
to various neurological and neurodegenerative disorders, including
stroke, Huntington's disease, Amyotrophic Lateral Sclerosis and
Alzheimer's disease. HDAC inhibition may induce the expression of
anti-mitotic and anti-apoptotic genes, such as p21 and HSP-70,
which facilitate survival. HDAC inhibitors can act on other neural
cell types in the central nervous system, such as reactive
astrocytes and microglia, to reduce inflammation and secondary
damage during neuronal injury or disease. HDAC inhibition is a
promising therapeutic approach for the treatment of a range of
central nervous system disorders (Langley B et al., 2005, Current
Drug Targets--CNS & Neurological Disorders, 4: 41-50).
[0010] Histone deacetylase is known to play an essential role in
the transcriptional machinery for regulating gene expression,
induce histone hyperacetylation and to affect the gene expression.
Therefore, it is useful as a therapeutic or prophylactic agent for
diseases caused by abnormal gene expression such as inflammatory
disorders, diabetes, diabetic complications, homozygous
thalassemia, fibrosis, cirrhosis, acute promyelocytic leukaemia
(APL), organ transplant rejections, autoimmune diseases, protozoal
infections, tumors, etc. Thus, there remains a need for the
development of novel inhibitors of histone deacetylases and tubulin
histone deacetylases. In particular, inhibitors that are more
potent and/or more specific for their particular target than known
HDAC and TDAC inhibitors are needed. HDAC inhibitors specific for a
certain class or member of the HDAC family would be particularly
useful both in the treatment of proliferative diseases and protein
deposition disorders and in the study of HDACs, particularly HDAC6.
Inhibitors that are specific for HDAC versus TDAC and vice versa
are also useful in treating disease and probing biological
pathways. The present invention provides novel compounds,
pharmaceutical compositions thereof, and methods of using these
compounds to treat disorders related to HDAC6 including cancers,
inflammatory, autoimmune, neurological and neurodegenerative
disorders.
SUMMARY OF THE INVENTION
[0011] In one aspect, the invention provides a compound of formula
I:
##STR00001##
[0012] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0013] each of X.sub.1, X.sub.2, X.sub.3, or X.sub.4 is
independently N or CR', wherein 2 of X.sub.1, X.sub.2, X.sub.3, or
X.sub.4 are N;
[0014] each R' is independently H, optionally substituted alkyl,
optionally substituted aryl, or halo:
[0015] R.sub.1 is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; and R.sub.2 is H, alkyl, haloalkyl,
alkenyl, arylalkyl, aryl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O)R.sub.5, each of which may be optionally
substituted;
[0016] p is 0, 1, or 2;
[0017] each R.sub.5 is independently H; optionally substituted
alkyl, optionally substituted carbocyclic, optionally substituted
heterocyclic, optionally substituted aryl, or optionally
substituted heteroaryl;
[0018] wherein R.sub.1 and R.sub.2 are both not simultaneously
H.
[0019] In another aspect, the invention provides a pharmaceutical
composition comprising a compound of any of the formulae herein
(e.g., formula I).
[0020] In another aspect, the invention provides a method of
selectively inhibiting HDAC6 over other HDACs in a subject,
comprising administering to the subject a compound of any of the
formulae herein (e.g., formula I).
[0021] In another aspect, the invention provides a method of
treating a disease mediated by HDAC6 in a subject comprising
administering to the subject a compound of any of the formulae
herein (e.g., formula I).
[0022] In another aspect, the invention provides a method of
treating a subject suffering from or susceptible to multiple
myeloma comprising administering to a subject in need thereof a
therapeutically effective amount of a compound of any of the
formulae herein (e.g., formula I), to thereby treat the subject
suffering from or susceptible to multiple myeloma.
[0023] In another aspect, the invention provides a kit comprising a
compound capable of inhibiting HDAC activity selected from one or
more compounds of any of the formulae herein (e.g., formula I); and
instructions for use in treating multiple myeloma.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0024] Listed below are definitions of various terms used to
describe this invention. These definitions apply to the terms as
they are used throughout this specification and claims, unless
otherwise limited in specific instances, either individually or as
part of a larger group.
[0025] The number of carbon atoms in a hydrocarbyl substituent can
be indicated by the prefix "C.sub.x-C.sub.y," where x is the
minimum and y is the maximum number of carbon atoms in the
substituent. Likewise, a C.sub.x chain means a hydrocarbyl chain
containing x carbon atoms.
[0026] If a linking element in a depicted structure is "absent" or
a "bond", then the left element in the depicted structure is
directly linked to the right element in the depicted structure. For
example, if a chemical structure is depicted as X-(L).sub.n-Y
wherein L is absent or n is 0, then the chemical structure is
X--Y.
[0027] The term "alkyl," as used herein, refers to saturated,
straight- or branched-chain hydrocarbon moieties containing, in
certain embodiments, between one and six, or one and eight carbon
atoms, respectively. Examples of C.sub.1-C.sub.6 alkyl moieties
include, but are not limited to, methyl, ethyl, propyl, isopropyl,
n-butyl, tert-butyl, neopentyl, n-hexyl moieties; and examples of
C.sub.1-C.sub.8 alkyl moieties include, but are not limited to,
methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl,
n-hexyl, heptyl, and octyl moieties.
[0028] The term "alkenyl," as used herein, denotes a monovalent
group derived from a hydrocarbon moiety containing, in certain
embodiments, from two to six, or two to eight carbon atoms having
at least one carbon-carbon double bond. The double bond may or may
not be the point of attachment to another group. Alkenyl groups
include, but are not limited to, for example, ethenyl, propenyl,
butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl and the like.
[0029] The term "alkynyl," as used herein, denotes a monovalent
group derived from a hydrocarbon moiety containing, in certain
embodiments, from two to six, or two to eight carbon atoms having
at least one carbon-carbon triple bond. The alkynyl group may or
may not be the point of attachment to another group. Representative
alkynyl groups include, but are not limited to, for example,
ethynyl, 1-propynyl, 1-butynyl, heptynyl, octynyl and the like.
[0030] The term "alkoxy" refers to an --O-alkyl moiety.
[0031] The term "aryl," as used herein, refers to a mono- or
poly-cyclic carbocyclic ring system having one or more aromatic
rings, fused or non-fused, including, but not limited to, phenyl,
naphthyl, tetrahydronaphthyl, indanyl, idenyl and the like.
[0032] The term "aralkyl," or "arylalkyl," as used herein, refers
to an alkyl residue attached to an aryl ring. Examples include, but
are not limited to, benzyl, phenethyl and the like.
[0033] The term "carbocyclic," as used herein, denotes a monovalent
group derived from a monocyclic or polycyclic saturated, partially
unsatured, or fully unsaturated carbocyclic ring compound. Examples
of carbocyclic groups include groups found in the cycloalkyl
definition and aryl definition.
[0034] The term "cycloalkyl," as used herein, denotes a monovalent
group derived from a monocyclic or polycyclic saturated or
partially unsatured carbocyclic ring compound. Examples of
C.sub.3-C.sub.8-cycloalkyl include, but not limited to,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl and
cyclooctyl; and examples of C.sub.3-C.sub.12-cycloalkyl include,
but not limited to, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, bicyclo [2.2.1] heptyl, and bicyclo [2.2.2] octyl. Also
contemplated are monovalent groups derived from a monocyclic or
polycyclic carbocyclic ring compound having at least one
carbon-carbon double bond by the removal of a single hydrogen atom.
Examples of such groups include, but are not limited to,
cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl,
cycloheptenyl, cyclooctenyl, and the like.
[0035] The term "heteroaryl," as used herein, refers to a mono- or
poly-cyclic (e.g., bi-, or tri-cyclic or more) fused or non-fused,
moieties or ring system having at least one aromatic ring, having
from five to ten ring atoms of which one ring atom is selected from
S, O and N; zero, one or two ring atoms are additional heteroatoms
independently selected from S, O and N; and the remaining ring
atoms are carbon. Heteroaryl includes, but is not limited to,
pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl,
thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl,
thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl,
benzooxazolyl, quinoxalinyl, and the like.
[0036] The term "heteroaralkyl," as used herein, refers to an alkyl
residue residue attached to a heteroaryl ring. Examples include,
but are not limited to, pyridinylmethyl, pyrimidinylethyl and the
like.
[0037] The term "heterocycloalkyl," as used herein, refers to a
non-aromatic 3-, 4-, 5-, 6- or 7-membered ring or a bi- or
tri-cyclic group fused of non-fused system, where (i) each ring
contains between one and three heteroatoms independently selected
from oxygen, sulfur and nitrogen, (ii) each 5-membered ring has 0
to 1 double bonds and each 6-membered ring has 0 to 2 double bonds,
(iii) the nitrogen and sulfur heteroatoms may optionally be
oxidized, (iv) the nitrogen heteroatom may optionally be
quaternized, and (iv) any of the above rings may be fused to a
benzene ring. Representative heterocycloalkyl groups include, but
are not limited to, [1,3]dioxolane, pyrrolidinyl, pyrazolinyl,
pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl,
piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl,
thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl.
[0038] The term "alkylamino" refers to a group having the structure
--NH(C.sub.1-C.sub.12 alkyl) where C.sub.1-C.sub.12 alkyl is as
previously defined.
[0039] The term "acyl" includes residues derived from acids,
including but not limited to carboxylic acids, carbamic acids,
carbonic acids, sulfonic acids, and phosphorous acids. Examples
include aliphatic carbonyls, aromatic carbonyls, aliphatic
sulfonyls, aromatic sulfinyls, aliphatic sulfinyls, aromatic
phosphates and aliphatic phosphates. Examples of aliphatic
carbonyls include, but are not limited to, acetyl, propionyl,
2-fluoroacetyl, butyryl, 2-hydroxy acetyl, and the like.
[0040] In accordance with the invention, any of the aryls,
substituted aryls, heteroaryls and substituted heteroaryls
described herein, can be any aromatic group. Aromatic groups can be
substituted or unsubstituted.
[0041] The terms "hal," "halo" and "halogen," as used herein, refer
to an atom selected from fluorine, chlorine, bromine and
iodine.
[0042] The term "oxo" as used herein, refers to an oxygen that is
attached to a carbon, preferably by a double bond (e.g.,
carbonyl).
[0043] As described herein, compounds of the invention may
optionally be substituted with one or more substituents, such as
are illustrated generally above, or as exemplified by particular
classes, subclasses, and species of the invention. It will be
appreciated that the phrase "optionally substituted" is used
interchangeably with the phrase "substituted or unsubstituted." In
general, the term "substituted", whether preceded by the term
"optionally" or not, refers to the replacement of hydrogen radicals
in a given structure with the radical of a specified substituent.
Unless otherwise indicated, an optionally substituted group may
have a substituent at each substitutable position of the group, and
when more than one position in any given structure may be
substituted with more than one substituent selected from a
specified group, the substituent may be either the same or
different at every position. The terms "optionally substituted",
"optionally substituted alkyl," "optionally substituted"
"optionally substituted alkenyl," "optionally substituted alkynyl",
"optionally substituted cycloalkyl," "optionally substituted
cycloalkenyl," "optionally substituted aryl", "optionally
substituted heteroaryl," "optionally substituted aralkyl",
"optionally substituted heteroaralkyl," "optionally substituted
heterocycloalkyl," and any other optionally substituted group as
used herein, refer to groups that are substituted or unsubstituted
by independent replacement of one, two, or three or more of the
hydrogen atoms thereon with substituents including, but not limited
to:
[0044] alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocycloalkyl,
heteroaryl, arylalkyl, heteroarylalkyl, haloalkyl (e.g.,
--CF.sub.3), haloalkoxy (e.g., --OCF.sub.1),
[0045] --F, --Cl, --Br, --I,
[0046] --OH, protected hydroxy, oxygen, oxo,
[0047] --NO.sub.2, --CN,
[0048] --NH.sub.2, protected amino, --NH--C.sub.1-C.sub.12-alkyl,
--NH-aryl, -dialkylamino,
[0049] --O--C.sub.1-C.sub.12-alkyl, --O-aryl,
[0050] --C(O)--, --C(O)O--, --C(O)NH--, --OC(O)--, --OC(O)O--,
--OC(O)NH--, --NHC(O)--, --NHC(O)O--,
--C(O)--C.sub.1-C.sub.12-alkyl,
--C(O)--C.sub.3-C.sub.12-cycloalkyl, --C(O)-aryl,
--C(O)-heteroaryl, --C(O)-heterocycloalkyl,
[0051] --C(O)O--C.sub.1-C.sub.12-alkyl,
--C(O)O--C.sub.3-C.sub.12-cycloalkyl, --C(O)O-aryl,
--C(O)O-heteroaryl, --C(O)O-heterocycloalkyl,
[0052] --CONH.sub.2, --CONH--C.sub.1-C.sub.12-alkyl,
--CONH-aryl,
[0053] --OCO.sub.2--C.sub.1-C.sub.12-alkyl, --OCO.sub.2-aryl,
--OCONH.sub.2, --OCONH--C.sub.1-C.sub.12-alkyl, --OCONH-aryl,
[0054] --NHC(O)--C.sub.1-C.sub.12-alkyl, --NHC(O)-aryl,
--NHCO.sub.2--C.sub.1-C.sub.12-alkyl, --NHCO.sub.2-aryl, [0055]
--S(O)--C.sub.1-C.sub.12-alkyl, --S(O)-aryl,
--SO.sub.2NH--C.sub.1-C.sub.12-alkyl, --SO.sub.2NH-aryl,
[0056] --NHSO.sub.2--C.sub.1-C.sub.12-alkyl, --NHSO.sub.2-aryl,
or
[0057] --SH, --S--C.sub.1-C.sub.12-alkyl, or --S-aryl.
[0058] In certain embodiments, the optionally substituted groups
include the following: C.sub.1-C.sub.12-alkyl,
C.sub.2-C.sub.12-alkenyl, C.sub.2-C.sub.12-alkynyl,
C.sub.3-C.sub.12-cycloalkyl, C.sub.3-C.sub.12-aryl,
C.sub.3-C.sub.12-heterocycloalkyl, C.sub.3-C.sub.12-heteroaryl,
C.sub.4-C.sub.12-arylalkyl, or
C.sub.2-C.sub.12-heteroarylalkyl.
[0059] It is understood that the aryls, heteroaryls, alkyls, and
the like can be further substituted.
[0060] As used herein, the term "metal chelator" refers to any
molecule or moiety that is capable of forming a complex (i.e.,
"chelates") with a metal ion. In certain exemplary embodiments, a
metal chelator refers to to any molecule or moiety that "binds" to
a metal ion, in solution, making it unavailable for use in
chemical/enzymatic reactions. In certain embodiments, the solution
comprises aqueous environments under physiological conditions.
Examples of metal ions include, but are not limited to, Ca.sup.2+,
Fe.sup.3+, Zn.sup.2+, Na.sup.+, etc. In certain embodiments, the
metal chelator binds Zn.sup.2+. In certain embodiments, molecules
of moieties that precipitate metal ions are not considered to be
metal chelators.
[0061] As used herein, the term "small molecule" refers to a
non-peptidic, non-oligomeric organic compound either synthesized in
the laboratory or found in nature. Small molecules, as used herein,
can refer to compounds that are "natural product-like", however,
the term "small molecule" is not limited to "natural product-like"
compounds. Rather, a small molecule is typically characterized in
that it contains several carbon-carbon bonds, and has a molecular
weight of less than 1500, although this characterization is not
intended to be limiting for the purposes of the present invention.
Examples of "small molecules" that occur in nature include, but are
not limited to, taxol, dynemicin, and rapamycin. In certain other
preferred embodiments, natural-product-like small molecules are
utilized.
[0062] The term "subject" as used herein refers to a mammal. A
subject therefore refers to, for example, dogs, cats, horses, cows,
pigs, guinea pigs, and the like. Preferably the subject is a human.
When the subject is a human, the subject may be referred to herein
as a patient.
[0063] "Treat", "treating" and "treatment" refer to a method of
alleviating or abating a disease and/or its attendant symptoms.
[0064] As used herein, the term "pharmaceutically acceptable salt"
refers to those salts of the compounds formed by the process of the
present invention which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of humans
and lower animals without undue toxicity, irritation, allergic
response and the like, and are commensurate with a reasonable
benefit/risk ratio. Pharmaceutically acceptable salts are well
known in the art. For example, S. M. Berge, et al. describes
pharmaceutically acceptable salts in detail in J. Pharmaceutical
Sciences, 66: 1-19 (1977). The salts can be prepared in situ during
the final isolation and purification of the compounds of the
invention, or separately by reacting the free base function with a
suitable organic acid. Examples of pharmaceutically acceptable
include, but are not limited to, nontoxic acid addition salts are
salts of an amino group formed with inorganic acids such as
hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid
and perchloric acid or with organic acids such as acetic acid,
maleic acid, tartaric acid, citric acid, succinic acid or malonic
acid or by using other methods used in the art such as ion
exchange. Other pharmaceutically acceptable salts include, but are
not limited to, adipate, alginate, ascorbate, aspartate,
benzenesulfonate, benzoate, bisulfate, borate, butyrate,
camphorate, camphorsulfonate, citrate, cyclopentanepropionate,
digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,
glucoheptonate, glycerophosphate, gluconate, hemisulfate,
heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate,
lactobionate, lactate, laurate, lauryl sulfate, malate, maleate,
malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate,
nitrate, oleate, oxalate, palmitate, pamoate, pectinate,
persulfate, 3-phenylpropionate, phosphate, picrate, pivalate,
propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like.
Representative alkali or alkaline earth metal salts include sodium,
lithium, potassium, calcium, magnesium, and the like. Further
pharmaceutically acceptable salts include, when appropriate,
nontoxic ammonium, quaternary ammonium, and amine cations formed
using counterions such as halide, hydroxide, carboxylate, sulfate,
phosphate, nitrate, alkyl having from 1 to 6 carbon atoms,
sulfonate and aryl sulfonate.
[0065] As used herein, the term "pharmaceutically acceptable ester"
refers to esters of the compounds formed by the process of the
present invention which hydrolyze in vivo and include those that
break down readily in the human body to leave the parent compound
or a salt thereof. Suitable ester groups include, for example,
those derived from pharmaceutically acceptable aliphatic carboxylic
acids, particularly alkanoic, alkenoic, cycloalkanoic and
alkanedioic acids, in which each alkyl or alkenyl moiety
advantageously has not more than 6 carbon atoms. Examples of
particular esters include, but are not limited to, formates,
acetates, propionates, butyrates, acrylates and
ethylsuccinates.
[0066] The term "pharmaceutically acceptable prodrugs" as used
herein refers to those prodrugs of the compounds formed by the
process of the present invention which are, within the scope of
sound medical judgment, suitable for use in contact with the
tissues of humans and lower animals with undue toxicity,
irritation, allergic response, and the like, commensurate with a
reasonable benefit/risk ratio, and effective for their intended
use, as well as the zwitterionic forms, where possible, of the
compounds of the present invention. "Prodrug", as used herein means
a compound which is convertible in vivo by metabolic means (e.g. by
hydrolysis) to afford any compound delineated by the formulae of
the instant invention. Various forms of prodrugs are known in the
art, for example, as discussed in Bundgaard, (ed.), Design of
Prodrugs, Elsevier (1985); Widder, et al. (ed.), Methods in
Enzymology, vol. 4, Academic Press (1985); Krogsgaard-Larsen, et
al., (ed). "Design and Application of Prodrugs, Textbook of Drug
Design and Development, Chapter 5, 113-191 (1991); Bundgaard, et
al., Journal of Drug Deliver Reviews, 8:1-38(1992); Bundgaard, J.
of Pharmaceutical Sciences, 77:285 et seq. (1988); Higuchi and
Stella (eds.) Prodrugs as Novel Drug Delivery Systems, American
Chemical Society (1975); and Bernard Testa & Joachim Mayer,
"Hydrolysis In Drug And Prodrug Metabolism: Chemistry, Biochemistry
And Enzymology," John Wiley and Sons, Ltd. (2002).
[0067] This invention also encompasses pharmaceutical compositions
containing, and methods of treating disorders through
administering, pharmaceutically acceptable prodrugs of compounds of
the invention. For example, compounds of the invention having free
amino, amido, hydroxy or carboxylic groups can be converted into
prodrugs. Prodrugs include compounds wherein an amino acid residue,
or a polypeptide chain of two or more (e.g., two, three or four)
amino acid residues is covalently joined through an amide or ester
bond to a free amino, hydroxy or carboxylic acid group of compounds
of the invention. The amino acid residues include but are not
limited to the 20 naturally occurring amino acids commonly
designated by three letter symbols and also includes
4-hydroxyproline, hydroxyysine, demosine, isodemosine,
3-methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid,
citrulline, homocysteine, homoserine, ornithine and methionine
sulfone. Additional types of prodrugs are also encompassed. For
instance, free carboxyl groups can be derivatized as amides or
alkyl esters. Free hydroxy groups may be derivatized using groups
including but not limited to hemisuccinates, phosphate esters,
dimethylaminoacetates, and phosphoryloxymethyloxy carbonyls, as
outlined in Advanced Drug Delivery Reviews, 1996, 19, 1 15.
Carbamate prodrugs of hydroxy and amino groups are also included,
as are carbonate prodrugs, sulfonate esters and sulfate esters of
hydroxy groups. Derivatization of hydroxy groups as (acyloxy)methyl
and (acyloxy)ethyl ethers wherein the acyl group may be an alkyl
ester, optionally substituted with groups including but not limited
to ether, amine and carboxylic acid functionalities, or where the
acyl group is an amino acid ester as described above, are also
encompassed. Prodrugs of this type are described in J. Med. Chem.
1996, 39, 10. Free amines can also be derivatized as amides,
sulfonamides or phosphonamides. All of these prodrug moieties may
incorporate groups including but not limited to ether, amine and
carboxylic acid functionalities
[0068] Combinations of substituents and variables envisioned by
this invention are only those that result in the formation of
stable compounds. The term "stable", as used herein, refers to
compounds which possess stability sufficient to allow manufacture
and which maintains the integrity of the compound for a sufficient
period of time to be useful for the purposes detailed herein (e.g.,
therapeutic or prophylactic administration to a subject).
[0069] The terms "isolated," "purified," or "biologically pure"
refer to material that is substantially or essentially free from
components that normally accompany it as found in its native state.
Purity and homogeneity are typically determined using analytical
chemistry techniques such as polyacrylamide gel electrophoresis or
high performance liquid chromatography. Particularly, in
embodiments the compound is at least 85% pure, more preferably at
least 90% pure, more preferably at least 95% pure, and most
preferably at least 99% pure.
[0070] As used herein, the term "disease" includes any one or more
of the following autoimmune diseases or disorders: diabetes
mellitus, arthritis (including rheumatoid arthritis, juvenile
rheumatoid arthritis, osteoarthritis, psoriatic arthritis),
multiple sclerosis, myasthenia gravis, systemic lupus
erythematosis, autoimmune thyroiditis, dermatitis (including atopic
dermatitis and eczematous dermatitis), psoriasis, Sjogren's
Syndrome, including keratoconjunctivitis sicca secondary to
Sjogren's Syndrome, alopecia areata, allergic responses due to
arthropod bite reactions, Crohn's disease, aphthous ulcer, iritis,
conjunctivitis, keratoconjunctivitis, ulcerative colitis, asthma,
allergic asthma, cutaneous lupus erythematosus, scleroderma,
vaginitis, proctitis, drug eruptions, leprosy reversal reactions,
erythema nodosum leprosum, autoimmune uveitis, allergic
encephalomyelitis, acute necrotizing hemorrhagic encephalopathy,
idiopathic bilateral progressive sensorineural hearing loss,
aplastic anemia, pure red cell anemia, idiopathic thrombocytopenia,
polychondritis, Wegener's granulomatosis, chronic active hepatitis.
Stevens-Johnson syndrome, idiopathic sprue, lichen planus, Graves
ophthalmopathy, sarcoidosis, primary biliary cirrhosis, uveitis
posterior, and interstitial lung fibrosis.
[0071] In another embodiment, disease refers to any one of Wilson's
disease, spinocerebellar ataxia, prion disease, Parkinson's
disease, Huntington's disease, amytrophic lateral sclerosis,
amyloidosis, Alzheimer's disease, Alexander's disease, alcoholic
liver disease, cystic fibrosis, Pick's Disease, spinal muscular
dystrophy or Lewy body dementia.
[0072] "Disease" also includes any one of rheumatoid spondylitis;
post ischemic perfusion injury; inflammatory bowel disease; chronic
inflammatory pulmonary disease, eczema, asthma,
ischemia/reperfusion injury, acute respiratory distress syndrome,
infectious arthritis, progressive chronic arthritis, deforming
arthritis, traumatic arthritis, gouty arthritis, Reiter's syndrome,
acute synovitis and spondylitis, glomerulonephritis, hemolytic
anemia, aplastic anemia, neutropenia, host versus graft disease,
allograft rejection, chronic thyroiditis, Graves' disease, primary
binary cirrhosis, contact dermatitis, skin sunburns, chronic renal
insufficiency, Guillain-Barre syndrome, uveitis, otitis media,
periodontal disease, pulmonary interstitial fibrosis, bronchitis,
rhinitis, sinusitis, pneumoconiosis, pulmonary insufficiency
syndrome, pulmonary emphysema, pulmonary fibrosis, silicosis, or
chronic inflammatory pulmonary disease.
[0073] "Disease" also refers to any one of cancer, tumor growth,
cancer of the colon, breast, bone, brain and others (e.g.,
osteosarcoma, neuroblastoma, colon adenocarcinoma), chronic
myelogenous leukemia (CML), acute myeloid leukemia (AML), acute
promyelocytic leukemia (APL), cardiac cancer (e.g., sarcoma,
myxoma, rhabdomyoma, fibroma, lipoma and teratoma); lung cancer
(e.g., bronchogenic carcinoma, alveolar carcinoma, bronchial
adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma);
various gastrointestinal cancer (e.g., cancers of esophagus,
stomach, pancreas, small bowel, and large bowel); genitourinary
tract cancer (e.g., kidney, bladder and urethra, prostate, testis;
liver cancer (e.g., hepatoma, cholangiocarcinoma, hepatoblastoma,
angiosarcoma, hepatocellular adenoma, hemangioma); bone cancer
(e.g., osteogenic sarcoma, fibrosarcoma, malignant fibrous
histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma,
multiple myeloma, malignant giant cell tumor chordoma,
osteochronfroma, benign chondroma, chondroblastoma,
chondromyxofibroma, osteoid osteoma and giant cell tumors); cancers
of the nervous system (e.g., of the skull, meninges, brain, and
spinal cord); gynecological cancers (e.g., uterus, cervix, ovaries,
vulva, vagina); hematologic cancer (e.g., cancers relating to
blood, Hodgkin's disease, non-Hodgkin's lymphoma); skin cancer
(e.g., malignant melanoma, basal cell carcinoma, squamous cell
carcinoma, Karposi's sarcoma, moles dysplastic nevi, lipoma,
angioma, dermatofibroma, keloids, psoriasis); and cancers of the
adrenal glands (e.g., neuroblastoma).
[0074] Compounds of the Invention
[0075] In one aspect, the invention provides a compound of formula
I:
##STR00002##
[0076] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0077] each of X.sub.1, X.sub.2, X.sub.3, or X.sub.4 is
independently N or CR', wherein 2 of X.sub.1, X.sub.2, X.sub.3, or
X.sub.4 are N;
[0078] each R' is independently H, optionally substituted alkyl,
optionally substituted aryl, or halo;
[0079] R.sub.1 is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; and
[0080] R.sub.2 is H, alkyl, haloalkyl, alkenyl, arylalkyl, aryl,
heteroaryl, heterocyclic, carbocyclic, --C(O)R.sub.5, or
--S(O).sub.pR.sub.5, each of which may be optionally
substituted:
[0081] each p is independently 0, 1, or 2; and
[0082] each R.sub.5 is independently H; optionally substituted
alkyl, optionally substituted carbocyclic, optionally substituted
heterocyclic, optionally substituted aryl, or optionally
substituted heteroaryl;
[0083] wherein R.sub.1 and R.sub.2 are both not simultaneously
H.
[0084] In one embodiment, the ring comprising X.sub.1, X.sub.2,
X.sub.3, and X.sub.4 is selected from the following:
##STR00003##
[0085] In another embodiment, R.sub.2 is H, methyl, ethyl, propyl,
i-propyl, butyl, s-butyl, t-butyl, pentyl, hexyl, heptyl, phenyl,
naphthyl, anthracenyl, pyridinyl, pyrimidinyl, pyrazinyl, indolyl,
imidazolyl, oxazolyl, furyl, thienyl, thiophenyl, thiazolyl,
triazolyl, isoxazolyl, quinolinyl, pyrrolyl, pyrazolyl, or
5,6,7,8-tetrahydroisoquinoline; each of which may be optionally
substituted.
[0086] In another embodiment, R.sub.1 is H, optionally substituted
alkyl, optionally substituted aryl, optionally substituted
arylalkyl, optionally substituted heteroarylalkyl, or optionally
substituted heteroaryl.
[0087] In a further embodiment, R.sub.1 is H, methyl, ethyl,
propyl, i-propyl, butyl, i-butyl, t-butyl, pentyl, hexyl, phenyl,
benzyl, phenethyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, piperidinyl, pyrrolidinyl, tetrahydropyranyl,
thiophenyl, or tetrahydrofuranyl, each of which may be optionally
substituted.
[0088] In certain embodiments, R.sub.1 is substituted with 1-4
groups independently selected from optionally substituted alkyl,
optionally substituted aryl, optionally substituted heteroaryl,
optionally substituted heterocycloalkyl, halo, haloalkyl, alkoxy,
haloalkoxy, and nitro.
[0089] In a further embodiment, R.sub.1 is substituted with 1-4
groups independently selected from methyl, phenyl, pyridyl,
morpholino, indolyl, pyrazinyl, F, Cl, Br, methoxy, OCF.sub.3, and
trifluoromethyl; each of which may be further substituted.
[0090] In certain embodiments, each of the above substituents may
be optionally substituted by alkyl, aryl, haloalkyl, haloalkoxy,
halo, or alkoxy.
[0091] In another embodiment, R.sub.2 is H.
[0092] In one embodiment, the invention provides a compound of
formula II:
##STR00004##
[0093] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0094] each of X.sub.1, X.sub.2, or X.sub.3, is independently N or
CR', wherein 2 of X.sub.1, X.sub.2, or X.sub.3 are N;
[0095] R.sub.2 is H, an optionally substituted alkyl, an optionally
substituted aryl or an optionally substituted heteroaryl;
[0096] R.sub.1 is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, or carbocyclic, each of
which may be optionally substituted; and
[0097] each R' is independently H, optionally substituted alkyl,
optionally substituted aryl, or halo.
[0098] In certain embodiments, X.sub.2 and X.sub.3 are N.
[0099] In other embodiments, R.sub.2 is H, methyl, ethyl, propyl,
i-propyl, butyl, s-butyl, t-butyl, pentyl, hexyl, heptyl, phenyl,
pyridinyl, pyrimidinyl, or pyrazinyl; each of which may be
optionally substituted.
[0100] In a further embodiment, R.sub.2 is substituted by 1-4
groups independently selected from optionally substituted alkyl,
optionally substituted aryl, optionally substituted heteroaryl,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aralkyl, alkoxyl,
haloalkyl, halo, OH, NH.sub.2, NHR'', CN, N.sub.3, and NO.sub.2,
and R'' is H or alkyl.
[0101] In certain embodiments, R.sub.1 is H, methyl, ethyl, propyl,
i-propyl, butyl, i-butyl, t-butyl, pentyl, hexyl, phenyl, naphthyl,
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, piperidinyl,
pyrrolidinyl, tetrahydropyranyl, tetrahydrofuranyl, benzyl,
phenethyl, or pyridyl, each of which may be optionally
substituted.
[0102] In a further embodiment, R.sub.1 is substituted by 1-4
groups independently selected from optionally substituted alkyl,
optionally substituted aryl, optionally substituted heteroaryl,
optionally substituted cycloalkyl, optionally substituted
heterocycloalkyl, optionally substituted aralkyl, alkoxyl,
haloalkoxy, haloalkyl, halo, OH, NH.sub.2, NHR'', CN, N.sub.3, and
NO.sub.2, and R'' is H or alkyl.
[0103] In another embodiment, R.sub.2 is H.
[0104] In another embodiment, the invention provides a compound of
formula III:
##STR00005##
[0105] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0106] R.sub.2 is H, methyl, pyridyl, or phenyl, each of which may
be optionally substituted;
[0107] R.sub.1 is H, methyl, ethyl, cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, piperidinyl, pyrrolidinyl,
tetrahydropyranyl, tetrahydrofuranyl, benzyl, phenethyl, pyridyl,
or phenyl, each of which may be optionally substituted.
[0108] In one embodiment, R.sub.2 is optionally substituted with
1-4 groups independently selected from optionally substituted aryl,
optionally substituted heteroaryl, F, Cl, Br, methyl, ethyl,
propyl, methoxy, and CF.sub.3. In certain embodiments, each of the
above substituents may be optionally substituted.
[0109] In other embodiments, R.sub.1 is optionally substituted with
1-4 groups independently selected from methyl, phenyl, pyridyl,
morpholino, indolyl, pyrazinyl, F, Cl, Br, methoxy, OCF.sub.3,
trifluoromethyl, OH, NH.sub.2, CN, and NO.sub.2. In certain
embodiments, R.sub.1 is further substituted by alkyl, aryl,
haloalkyl, halo, or alkoxy. In certain embodiments, each of the
above substituents may be optionally substituted.
[0110] In another embodiment, R.sub.2 is H.
[0111] In other embodiments, R.sub.1 is phenyl or pyridyl, and
R.sub.2 is H or Me.
[0112] In a further embodiment, R.sub.1 is further substituted with
1-4 substituents independently selected from alkyl, halo, alkoxy,
haloalkyl, haloalkoxy, aryl, heteroaryl, carbocyclic, heterocyclic,
and NO.sub.2. In another further embodiment, R.sub.1 is further
substituted with 1-4 substituents independently selected from
methyl, F, Cl, OCH.sub.3, CF.sub.3, OCF.sub.3, phenyl, pyridyl,
morpholinyl, or NO.sub.2.
[0113] In one embodiment, the invention provides a compound of
formula IV:
##STR00006##
[0114] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0115] R.sub.2 is H, alkyl, haloalkyl, arylalkyl, aryl, heteroaryl,
heterocyclic, carbocyclic, --C(O)R.sub.5, or --S(O).sub.pR.sub.5,
each of which may be optionally substituted;
[0116] R.sub.x is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted;
[0117] R.sub.y is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; or
[0118] R.sub.x and R.sub.y together with the carbon to which each
is attached, forms a cycloalkyl or heterocycloalkyl, each of which
is optionally substituted:
[0119] each R.sub.A is independently alkyl, alkoxy, cycloalkyl,
aryl, heterocycloalkyl, heteroaryl, arylalkyl, heteroarylalkyl,
haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN, or --NH.sub.2;
or two R.sub.A groups together with the atoms to which each is
attached, can form an optionally substituted ring;
[0120] m is 0, 1, 2, 3, or 4; and
[0121] p is 0, 1, 2, or 3.
[0122] In another embodiment, the invention provides a compound of
formula V:
##STR00007##
[0123] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0124] R.sub.2 is H, alkyl, haloalkyl, arylalkyl, aryl, heteroaryl,
heterocyclic, carbocyclic, --C(O)R.sub.5, or --S(O).sub.pR.sub.5,
each of which may be optionally substituted;
[0125] R.sub.x is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted;
[0126] R.sub.y is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; or
[0127] R.sub.x and R.sub.y together with the carbon to which each
is attached, forms a cycloalkyl or heterocycloalkyl, each of which
is optionally substituted:
[0128] each R.sub.A is independently alkyl, alkoxy, cycloalkyl,
aryl, heterocycloalkyl, heteroaryl, arylalkyl, heteroarylalkyl,
haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN, or --NH.sub.2;
or two R.sub.A groups together with the atoms to which each is
attached, can form an optionally substituted ring; and
[0129] m is 0, 1, 2, 3, or 4.
[0130] In one embodiment, R.sub.2 is H or Me.
[0131] In other embodiments, R.sub.x is H, alkyl, or aryl; and
R.sub.y is H or alkyl; each of which is optionally substituted.
[0132] In certain embodiments, R.sub.x is H, methyl, ethyl, or
phenyl.
[0133] In a further embodiment, R.sub.y is H, methyl or ethyl.
[0134] In other further embodiments, R.sub.x and R.sub.y together
with the carbon to which each is attached, forms a cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl,
each of which is optionally substituted.
[0135] In certain embodiments, R.sub.x and R.sub.y together with
the carbon to which each is attached, forms tetrahydropyran,
piperidine, piperazine, morpholine, tetrahydrofuran,
tetrahydrothiophene, pyrrolidine, oxozolidine, or imidazolidine,
each of which is optionally substituted.
[0136] In still other embodiments, R.sub.A is H, alkyl, alkoxy,
cycloalkyl, aryl, heterocycloalkyl, heteroaryl, arylalkyl,
heteroarylalkyl, haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN,
or --NH.sub.2, and m is 0 or 1.
[0137] In another embodiment, the invention provides a compound of
formula VI:
##STR00008##
[0138] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0139] R.sub.x and R.sub.y together with the carbon to which each
is attached, forms a cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl, tetrahydropyran, piperidine,
piperazine, morpholine, tetrahydrofuran, tetrahydrothiophene,
pyrrolidine, oxozolidine, or imidazolidine, each of which is
optionally substituted;
[0140] each R.sub.A is independently alkyl, alkoxy, cycloalkyl,
aryl, heterocycloalkyl, heteroaryl, arylalkyl, heteroarylalkyl,
haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN, or --NH.sub.2;
or two R.sub.A groups together with the atoms to which each is
attached, can form an optionally substituted ring; and
[0141] m is 0, 1, or 2.
[0142] In one embodiment, R.sub.x and R.sub.y together with the
carbon to which each is attached, forms a cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, or tetrahydropyran.
[0143] In another embodiment, m is 1 or 2, and each R.sub.A is
independently methyl, phenyl, F, Cl, methoxy, OCF.sub.3, or
CF.sub.3; or two R.sub.A groups together with the atoms to which
each is attached, can form an optionally substituted cycloalkyl or
heterocyclic ring.
[0144] In other embodiments, m is 0.
[0145] In certain embodiments, the invention provides a compound of
formula VI:
##STR00009##
[0146] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0147] R.sub.x is H, alkyl, or aryl; each of which is optionally
substituted;
[0148] R.sub.y is H or alkyl; each of which is optionally
substituted;
[0149] each R.sub.A is independently alkyl, alkoxy, cycloalkyl,
aryl, heterocycloalkyl, heteroaryl, arylalkyl, heteroarylalkyl,
haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN, or --NH.sub.2;
or two R.sub.A groups together can form an optionally substituted
cycloalkyl or heterocyclic ring; and
[0150] m is 0, 1, or 2.
[0151] In some embodiments, R.sub.x is H, methyl, ethyl, or
phenyl.
[0152] In other embodiments, R.sub.y is H, methyl or ethyl.
[0153] In certain embodiments, m is 1 or 2, and each R.sub.A is
independently methyl, phenyl, F, Cl, methoxyl, or CF.sub.3; or two
R.sub.A groups together can form an optionally substituted
cycloalkyl or heterocyclic ring.
[0154] In other embodiments, m is 0.
[0155] In another embodiment, the invention provides a compound of
formula VII:
##STR00010##
[0156] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0157] ring A is a heteroaryl or heterocyclic;
[0158] R.sub.2 is H, alkyl, haloalkyl, arylalkyl, aryl, heteroaryl,
heterocyclic, carbocyclic, --C(O)R.sub.5, or --S(O).sub.pR.sub.5,
each of which may be optionally substituted;
[0159] R.sub.x is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted;
[0160] R.sub.y is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; or
[0161] R.sub.x and R.sub.y together with the carbon to which each
is attached, forms a cycloalkyl or heterocycloalkyl, each of which
is optionally substituted:
[0162] each R.sub.A is independently alkyl, alkoxy, cycloalkyl,
aryl, heterocycloalkyl, heteroaryl, arylalkyl, heteroarylalkyl,
haloalkyl, haloalkoxy, halo, OH, --NO.sub.2. --CN, or --NH.sub.2;
or two R.sub.A groups together can form an optionally substituted
cycloalkyl or heterocyclic ring;
[0163] m is 0, 1, 2, 3, or 4; and
[0164] p is 0, 1, 2, or 3.
[0165] In another embodiment, the invention provides a compound of
formula VII-A:
##STR00011##
[0166] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0167] ring A is a heteroaryl or heterocyclic;
[0168] R.sub.2 is H, alkyl, haloalkyl, arylalkyl, aryl, heteroaryl,
heterocyclic, carbocyclic, --C(O)R.sub.5, or --S(O).sub.pR.sub.5,
each of which may be optionally substituted;
[0169] R.sub.x is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted;
[0170] R.sub.y is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; or
[0171] R.sub.x and R.sub.y together with the carbon to which each
is attached, forms a cycloalkyl or heterocycloalkyl, each of which
is optionally substituted:
[0172] each R.sub.A is independently alkyl, alkoxy, cycloalkyl,
aryl, heterocycloalkyl, heteroaryl, arylalkyl, heteroarylalkyl,
haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN, or --NH.sub.2;
or two R.sub.A groups together can form an optionally substituted
cycloalkyl or heterocyclic ring; and
[0173] m is 0, 1, 2, 3, or 4.
[0174] In certain embodiments, ring A is acridinyl, azocinyl,
benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl,
benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl,
benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl,
NH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl,
decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,
dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl,
imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl,
indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl,
isochromanyl, isoindazolyl, isoindolinyl, isoindolyl,
isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl,
naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl,
1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl; 1,2,5-oxadiazolyl,
1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl,
pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl,
phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl,
piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl,
pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole,
pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl,
pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl,
quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,
tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,
6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl,
1,2,5-thiadiazolyl, 1,3,4thiadiazolyl, thianthrenyl, thiazolyl,
thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl,
thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl,
1,2,5-triazolyl, 1,3,4-triazolyl, or xanthenyl.
[0175] In other embodiments, R.sub.2 is H or Me.
[0176] In various embodiments, R.sub.x is H, alkyl, aryl, or
heteroaryl.
[0177] In certain embodiments, R.sub.y is H, alkyl, aryl, or
heteroaryl.
[0178] In various embodiments, R.sub.y is H, methyl or pyridine,
and R.sub.y is H.
[0179] In certain embodiments, R.sub.x and R.sub.y together with
the carbon to which each is attached, forms a cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl,
each of which is optionally substituted.
[0180] In another embodiment, R.sub.x and R.sub.y together with the
carbon to which each is attached, forms tetrahydropyran,
piperidine, piperazine, morpholine, tetrahydrofuran,
tetrahydrothiophene, pyrrolidine, oxozolidine, or imidazolidine,
each of which is optionally substituted.
[0181] In various embodiments, R.sub.A is H, alkyl, alkoxy,
cycloalkyl, aryl, heterocycloalkyl, heteroaryl, arylalkyl,
heteroarylalkyl, haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN,
or --NH.sub.2, and m is 0 or 1.
[0182] In another embodiment, the invention provides a compound of
formula VIII:
##STR00012##
[0183] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0184] ring A is benzofuranyl, benzothiofuranyl, furanyl,
imidazolinyl, imidazolyl, isoquinolinyl, morpholinyl, oxazolidinyl,
pyrimidinyl, piperazinyl, piperidinyl, pyridinyl, pyridyl,
pyrazinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl,
pyrrolyl, quinazolinyl, quinolinyl, tetrahydrofuranyl,
tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiophenyl, or
triazinyl;
[0185] R.sub.x and R.sub.y together with the carbon to which each
is attached, forms a cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl, tetrahydropyran, piperidine,
piperazine, morpholine, tetrahydrofuran, tetrahydrothiophene,
pyrrolidine, oxozolidine, or imidazolidine, each of which is
optionally substituted;
[0186] each R.sub.A is independently alkyl, alkoxy, cycloalkyl,
aryl, heterocycloalkyl, heteroaryl, arylalkyl, heteroarylalkyl,
haloalkyl, haloalkoxy, halo, OH, --NO.sub.2, --CN, or --NH.sub.2;
or two R.sub.A groups together can form an optionally substituted
cycloalkyl or heterocyclic ring; and
[0187] m is 0, 1, or 2.
[0188] In certain embodiments, ring A is benzofuranyl,
benzothiofuranyl, furanyl, pyridyl, pyrazinyl, pyrimidinyl, or
thiophenyl.
[0189] In other embodiments, R.sub.x and R.sub.y together with the
carbon to which each is attached, forms a cyclopropyl, cyclopentyl,
cyclohexyl, or tetrahydropyran.
[0190] In certain embodiments, m is 1, and R.sub.A is F, Cl,
methoxyl, or CF.sub.3.
[0191] In other embodiments, m is 0.
[0192] Representative compounds of the invention include, but are
not limited to, the following compounds of Table 1 below.
TABLE-US-00001 TABLE 1 ##STR00013## 1 ##STR00014## 2 ##STR00015## 3
##STR00016## 4 ##STR00017## 5 ##STR00018## 6 ##STR00019## 7
##STR00020## 8 ##STR00021## 9 ##STR00022## 10 ##STR00023## 11
##STR00024## 12 ##STR00025## 13 ##STR00026## 14 ##STR00027## 15
##STR00028## 16 ##STR00029## 17 ##STR00030## 18 ##STR00031## 19
##STR00032## 20 ##STR00033## 21 ##STR00034## 22 ##STR00035## 23
##STR00036## 24 ##STR00037## 25 ##STR00038## 26 ##STR00039## 27
##STR00040## 28 ##STR00041## 29 ##STR00042## 30 ##STR00043## 31
##STR00044## 32 ##STR00045## 33 ##STR00046## 34 ##STR00047## 35
##STR00048## 36 ##STR00049## 37 ##STR00050## 38 ##STR00051## 39
##STR00052## 40 ##STR00053## 41 ##STR00054## 42 ##STR00055## 43
##STR00056## 44 ##STR00057## 45 ##STR00058## 46 ##STR00059## 47
##STR00060## 48 ##STR00061## 49 ##STR00062## 50 ##STR00063## 51
##STR00064## 52 ##STR00065## 53 ##STR00066## 54 ##STR00067## 55
##STR00068## 57 ##STR00069## 58 ##STR00070## 59 ##STR00071## 60
##STR00072## 61 ##STR00073## 62 ##STR00074## 63 ##STR00075## 65
##STR00076## 66 ##STR00077## 67 ##STR00078## 68 ##STR00079## 69
##STR00080## 70 ##STR00081## 71 ##STR00082## 72 ##STR00083## 73
##STR00084## 74 ##STR00085## 75 ##STR00086## 76 ##STR00087## 77
##STR00088## 78 ##STR00089## 79 ##STR00090## 80 ##STR00091## 81
##STR00092## 82 ##STR00093## 83 ##STR00094## 84 ##STR00095## 85
##STR00096## 86 ##STR00097## 97 ##STR00098## 87 ##STR00099## 88
##STR00100## 89 ##STR00101## 90 ##STR00102## 91 ##STR00103## 92
##STR00104## 93 ##STR00105## 94 ##STR00106## 95 ##STR00107## 96
##STR00108## 98 ##STR00109## 99 ##STR00110## 100 ##STR00111## 101
##STR00112## 102 ##STR00113## 103 ##STR00114## 104 ##STR00115## 105
##STR00116## 106 ##STR00117## 107 ##STR00118## 108 ##STR00119## 109
##STR00120## 110 ##STR00121## 111 ##STR00122## 112 ##STR00123## 113
##STR00124## 114 ##STR00125## 115 ##STR00126## 116 ##STR00127## 117
##STR00128## 118 ##STR00129## 119 ##STR00130## 120 ##STR00131## 121
##STR00132## 122 ##STR00133## 123 ##STR00134## 124 ##STR00135## 125
##STR00136## 126
##STR00137## 127 ##STR00138## 128 ##STR00139## 129 ##STR00140## 130
##STR00141## 131 ##STR00142## 132 ##STR00143## 133 ##STR00144## 134
##STR00145## 135 ##STR00146## 136 ##STR00147## 137 ##STR00148## 138
##STR00149## 139 ##STR00150## 140 ##STR00151## 141 ##STR00152## 142
##STR00153## 143 ##STR00154## 144 ##STR00155## 145 ##STR00156## 146
##STR00157## 147 ##STR00158## 148 ##STR00159## 149 ##STR00160## 150
##STR00161## 151 ##STR00162## 152 ##STR00163## 153 ##STR00164## 154
##STR00165## 155
[0193] In certain embodiments, the compound of the invention is
selected from Table 2;
TABLE-US-00002 TABLE 2 ##STR00166## 73 ##STR00167## 81 ##STR00168##
82 ##STR00169## 84 ##STR00170## 97 ##STR00171## 87 ##STR00172## 88
##STR00173## 90 ##STR00174## 91 ##STR00175## 93 ##STR00176## 94
##STR00177## 95
[0194] In another aspect, the invention provides a pharmaceutical
composition comprising a compound of any of the formulae herein,
(e.g., formula I):
##STR00178##
[0195] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0196] each of X.sub.1, X.sub.2, X.sub.3, or X.sub.4 is
independently N or CR', wherein 2 of X.sub.1, X.sub.2, X.sub.3, or
X.sub.4 are N;
[0197] each R' is independently H, optionally substituted alkyl,
optionally substituted aryl, or halo;
[0198] R.sub.1 is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; and
[0199] R.sub.2 is H, alkyl, haloalkyl, alkenyl, arylalkyl, aryl,
heteroaryl, heterocyclic, carbocyclic, --C(O)R.sub.5, or
--S(O).sub.pR.sub.5, each of which may be optionally
substituted;
[0200] p is 0, 1, or 2;
[0201] each R.sub.5 is independently H; optionally substituted
alkyl, optionally substituted carbocyclic, optionally substituted
heterocyclic, optionally substituted aryl, or optionally
substituted heteroaryl;
[0202] wherein R.sub.1 and R.sub.2 are both not simultaneously H;
together with a pharmaceutically acceptable carrier.
[0203] In preferred embodiments, a compound useful in the invention
has one or more of the following properties: the compound is
capable of inhibiting at least one histone deacetylase; the
compound is capable of inhibiting HDAC6: the compound is a
selective HDAC6 inhibitor: the compound binds to the poly-ubiquitin
binding domain of HDAC6; the compound is capable of inducing
apoptosis in cancer cells (especially multiple myeloma cells,
non-Hodgkin's lymphoma (NML) cells, breast cancer cells, acute
myelogeous leukemia (AML) cells); and/or the compound is capable of
inhibiting aggresome formation.
[0204] In certain preferred embodiments, a compound of the
invention comprises a metal binding moiety, preferably a
zinc-binding moiety such as a hydroxamate. As noted above, certain
hydroxamates are potent inhibitors of HDAC6 activity; without
wishing to be bound by theory, it is believed that the potency of
these hydroxamates is due, at least in part, to the ability of the
compounds to bind zinc. In preferred embodiments, a compound of the
invention includes at least one portion or region which can confer
selectivity for a biological target implicated in the aggresome
pathway, e.g., a biological target having tubulin deacetylase
(TDAC) or HDAC activity, e.g., HDAC6. Thus, in certain preferred
embodiments, a compound of the invention includes a zinc-binding
moiety spaced from other portions of the molecule which are
responsible for binding to the biological target.
[0205] The invention also provides for a pharmaceutical composition
comprising a compound of any of the formulae herein (e.g., formula
I), or a pharmaceutically acceptable ester, salt, or prodrug
thereof, together with a pharmaceutically acceptable carrier.
[0206] Another object of the present invention is the use of a
compound as described herein (e.g., of any formulae herein) in the
manufacture of a medicament for use in the treatment of a disorder
or disease herein. Another object of the present invention is the
use of a compound as described herein (e.g., of any formulae
herein) for use in the treatment of a disorder or disease
herein.
[0207] In another aspect, the invention provides a method of
synthesizing a compound of any of the formulae herein (e.g.,
formula I). The synthesis of the compounds of the invention can be
found in the Examples below.
[0208] Another embodiment is a method of making a compound of any
of the formulae herein using any one, or combination of, reactions
delineated herein. The method can include the use of one or more
intermediates or chemical reagents delineated herein.
[0209] Another aspect is an isotopically labeled compound of any of
the formulae delineated herein. Such compounds have one or more
isotope atoms which may or may not be radioactive (e.g., .sup.1H,
.sup.2H, .sup.14C, .sup.13C, .sup.35S, .sup.32P, .sup.125I, and
.sup.131I) introduced into the compound. Such compounds are useful
for drug metabolism studies and diagnostics, as well as therapeutic
applications.
[0210] A compound of the invention can be prepared as a
pharmaceutically acceptable acid addition salt by reacting the free
base form of the compound with a pharmaceutically acceptable
inorganic or organic acid. Alternatively, a pharmaceutically
acceptable base addition salt of a compound of the invention can be
prepared by reacting the free acid form of the compound with a
pharmaceutically acceptable inorganic or organic base.
[0211] Alternatively, the salt forms of the compounds of the
invention can be prepared using salts of the starting materials or
intermediates.
[0212] The free acid or free base forms of the compounds of the
invention can be prepared from the corresponding base addition salt
or acid addition salt from, respectively. For example a compound of
the invention in an acid addition salt form can be converted to the
corresponding free base by treating with a suitable base (e.g.,
ammonium hydroxide solution, sodium hydroxide, and the like). A
compound of the invention in a base addition salt form can be
converted to the corresponding free acid by treating with a
suitable acid (e.g., hydrochloric acid, etc.).
[0213] Prodrug derivatives of the compounds of the invention can be
prepared by methods known to those of ordinary skill in the art
(e.g., for further details see Saulnier et al., (1994), Bioorganic
and Medicinal Chemistry Letters, Vol. 4, p. 1985). For example,
appropriate prodrugs can be prepared by reacting a non-derivatized
compound of the invention with a suitable carbamylating agent
(e.g., 1,1-acyloxyalkylcarbanochloridate, para-nitrophenyl
carbonate, or the like).
[0214] Protected derivatives of the compounds of the invention can
be made by means known to those of ordinary skill in the art. A
detailed description of techniques applicable to the creation of
protecting groups and their removal can be found in T. W. Greene,
"Protecting Groups in Organic Chemistry", 3rd edition, John Wiley
and Sons, Inc., 1999, and subsequent editions thereof.
[0215] Compounds of the present invention can be conveniently
prepared, or formed during the process of the invention, as
solvates (e.g., hydrates). Hydrates of compounds of the present
invention can be conveniently prepared by recrystallization from an
aqueous/organic solvent mixture, using organic solvents such as
dioxan, tetrahydrofuran or methanol.
[0216] Acids and bases useful in the methods herein are known in
the art. Acid catalysts are any acidic chemical, which can be
inorganic (e.g., hydrochloric, sulfuric, nitric acids, aluminum
trichloride) or organic (e.g., camphorsulfonic acid,
p-toluenesulfonic acid, acetic acid, ytterbium triflate) in nature.
Acids are useful in either catalytic or stoichiometric amounts to
facilitate chemical reactions. Bases are any basic chemical, which
can be inorganic (e.g., sodium bicarbonate, potassium hydroxide) or
organic (e.g., triethylamine, pyridine) in nature. Bases are useful
in either catalytic or stoichiometric amounts to facilitate
chemical reactions.
[0217] In addition, some of the compounds of this invention have
one or more double bonds, or one or more asymmetric centers. Such
compounds can occur as racemates, racemic mixtures, single
enantiomers, individual diastereomers, diastereomeric mixtures, and
cis- or trans- or E- or Z-double isomeric forms, and other
stereoisomeric forms that may be defined, in terms of absolute
stereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino
acids. All such isomeric forms of these compounds are expressly
included in the present invention. Optical isomers may be prepared
from their respective optically active precursors by the procedures
described above, or by resolving the racemic mixtures. The
resolution can be carried out in the presence of a resolving agent,
by chromatography or by repeated crystallization or by some
combination of these techniques which are known to those skilled in
the art. Further details regarding resolutions can be found in
Jacques, et al., Enantiomers, Racemates, and Resolutions (John
Wiley & Sons, 1981). The compounds of this invention may also
be represented in multiple tautomeric forms, in such instances, the
invention expressly includes all tautomeric forms of the compounds
described herein. When the compounds described herein contain
olefinic double bonds or other centers of geometric asymmetry, and
unless specified otherwise, it is intended that the compounds
include both E and Z geometric isomers. Likewise, all tautomeric
forms are also intended to be included. The configuration of any
carbon-carbon double bond appearing herein is selected for
convenience only and is not intended to designate a particular
configuration unless the text so states; thus a carbon-carbon
double bond depicted arbitrarily herein as trans may be cis, trans,
or a mixture of the two in any proportion. All such isomeric forms
of such compounds are expressly included in the present invention.
All crystal forms of the compounds described herein are expressly
included in the present invention.
[0218] The synthesized compounds can be separated from a reaction
mixture and further purified by a method such as column
chromatography, high pressure liquid chromatography, or
recrystallization. As can be appreciated by the skilled artisan,
further methods of synthesizing the compounds of the formulae
herein will be evident to those of ordinary skill in the art.
Additionally, the various synthetic steps may be performed in an
alternate sequence or order to give the desired compounds. In
addition, the solvents, temperatures, reaction durations, etc.
delineated herein are for purposes of illustration only and one of
ordinary skill in the art will recognize that variation of the
reaction conditions can produce the desired compounds of the
present invention. Synthetic chemistry transformations and
protecting group methodologies (protection and deprotection) useful
in synthesizing the compounds described herein are known in the art
and include, for example, those such as described in R. Larock,
Comprehensive Organic Transformations, VCH Publishers (1989): T. W.
Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis,
2d. Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser,
Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and
Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for
Organic Synthesis, John Wiley and Sons (1995), and subsequent
editions thereof.
[0219] In embodiments, the invention provides for the intermediate
compounds of the formulae delineated herein and methods of
converting such compounds to compounds of the formulae herein
(e.g., in schemes herein) comprising reacting a compound herein
with one or more reagents in one or more chemical transformations
(including those provided herein) to thereby provide the compound
of any of the formulae herein or an intermediate compound
thereof.
[0220] The synthetic methods described herein may also additionally
include steps, either before or after any of the steps described in
any scheme, to add or remove suitable protecting groups in order to
ultimately allow synthesis of the compound of the formulae
described herein. The methods delineated herein contemplate
converting compounds of one formula to compounds of another formula
(e.g., in Scheme A, A1 to A2; A2 to A3; A1 to A3). The process of
converting refers to one or more chemical transformations, which
can be performed in situ, or with isolation of intermediate
compounds. The transformations can include reacting the starting
compounds or intermediates with additional reagents using
techniques and protocols known in the art, including those in the
references cited herein. Intermediates can be used with or without
purification (e.g., filtration, distillation, sublimation,
crystallization, trituration, solid phase extraction, and
chromatography).
##STR00179##
[0221] The compounds of this invention may be modified by appending
various functionalities via any synthetic means delineated herein
to enhance selective biological properties. Such modifications are
known in the art and include those which increase biological
penetration into a given biological system (e.g., blood, lymphatic
system, central nervous system), increase oral availability,
increase solubility to allow administration by injection, alter
metabolism and alter rate of excretion.
[0222] The compounds of the invention are defined herein by their
chemical structures and/or chemical names. Where a compound is
referred to by both a chemical structure and a chemical name, and
the chemical structure and chemical name conflict, the chemical
structure is determinative of the compound's identity.
[0223] The recitation of a listing of chemical groups in any
definition of a variable herein includes definitions of that
variable as any single group or combination of listed groups. The
recitation of an embodiment herein includes that embodiment as any
single embodiment or in combination with any other embodiments or
portions thereof. The recitation of an embodiment for a variable
herein includes that embodiment as any single embodiment or in
combination with any other embodiments or portions thereof.
METHODS OF THE INVENTION
[0224] In another aspect, the invention provides a method of
selectively inhibiting HDAC6 over other HDACs (e.g. HDAC1, HDAC2,
HDAC3) in a subject, comprising administering to the subject a
compound of formula I:
##STR00180##
[0225] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0226] each of X.sub.1, X.sub.2, X.sub.3, or X.sub.4 is
independently N or CR', wherein 2 of X.sub.1, X.sub.2, X.sub.3, or
X.sub.4 are N;
[0227] each R' is independently H, optionally substituted alkyl,
optionally substituted aryl, or halo;
[0228] R.sub.1 is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; and
[0229] R.sub.2 is H, alkyl, haloalkyl, alkenyl, arylalkyl, aryl,
heteroaryl, heterocyclic, carbocyclic, --C(O)R.sub.5, or
--S(O).sub.pR.sub.5, each of which may be optionally
substituted:
[0230] p is 0, 1, or 2;
[0231] each R.sub.5 is independently H; optionally substituted
alkyl, optionally substituted carbocyclic, optionally substituted
heterocyclic, optionally substituted aryl, or optionally
substituted heteroaryl;
[0232] wherein R.sub.1 and R.sub.2 are both not simultaneously
H.
[0233] In one embodiment, the compound of any of the formulae
herein (e.g., formula I) has a selectivity for HDAC6 of 5 to 1000
fold over other HDACs.
[0234] In another embodiment, the compound of of any of the
formulae herein (e.g., formula I) has a selectivity for HDAC6 when
tested in a HDAC enzyme assay of about 5 to 1000 fold over other
HDACs.
[0235] In certain embodiments, the compound has a selectivity for
HDAC6 of 15 to 40 fold over other HDACs.
[0236] In another aspect, the invention provides a method of
treating a disease mediated by HDAC6 in a subject comprising
administering to the subject a compound of formula I:
##STR00181##
[0237] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0238] each of X.sub.1, X.sub.2, X.sub.3, or X.sub.4 is
independently N or CR', wherein 2 of X.sub.1, X.sub.2, X.sub.3, or
X.sub.4 are N;
[0239] each R' is independently H, optionally substituted alkyl,
optionally substituted aryl, or halo;
[0240] R.sub.1 is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; and
[0241] R.sub.2 is H, alkyl, haloalkyl, alkenyl, arylalkyl, aryl,
heteroaryl, heterocyclic, carbocyclic, --C(O)R.sub.5, or
--S(O).sub.pR.sub.5, each of which may be optionally
substituted:
[0242] p is 0, 1, or 2;
[0243] each R.sub.5 is independently H; optionally substituted
alkyl, optionally substituted carbocyclic, optionally substituted
heterocyclic, optionally substituted aryl, or optionally
substituted heteroaryl;
[0244] wherein R.sub.1 and R.sub.2 are both not simultaneously
H.
[0245] In certain embodiments, the disease is cancer or a
proliferation disease.
[0246] In a further embodiment, the disease is lung cancer, colon
cancer, breast cancer, prostate cancer, liver cancer, brain cancer,
kidney cancer, ovarian cancer, stomach cancer, skin cancer, bone
cancer, gastric cancer, pancreatic cancer, glioma, gliobastoma,
hepatocellular carcinoma, papillary renal carcinoma, head and neck
squamous cell carcinoma, leukemias, lymphomas, myelomas, and solid
tumors.
[0247] In a further embodiment, the cancer is multiple myeloma.
[0248] In other embodiments, the disease is Wilson's disease,
spinocerebellar ataxia, prion disease, Parkinson's disease,
Huntington's disease, amytrophic lateral sclerosis, amyloidosis,
Alzheimer's disease, Alexander's disease, alcoholic liver disease,
cystic fibrosis, Pick's Disease, spinal muscular dystrophy or Lewy
body dementia.
[0249] In other embodiments, the disease is an inflammatory, immune
and autoimmune diseases including, but not limited to, arthritic
conditions, such as, rheumatoid arthritis, osteoarthritis; juvenile
arthritis, rheumatoid spondylitis; psoriasis; post ischemic
perfusion injury; inflammatory bowel disease; chronic inflammatory
pulmonary disease, eczema, asthma, psoriasis, ischemia/reperfusion
injury, ulcerative colitis, acute respiratory distress syndrome,
psoriatic arthritis, infectious arthritis, progressive chronic
arthritis, deforming arthritis, osteoarthritis, traumatic
arthritis, gouty arthritis, Reiter's syndrome, polychondritis,
acute synovitis and spondylitis, glomerulonephritis (with or
without nephrotic syndrome), autoimmune hematologic disorders (e.g.
hemolytic anemia, aplastic anemia, idiopathic thrombocytopenia and
neutropenia) ulcerative colitis, Crohn's disease, host versus graft
disease, graft versus host disease, allograft rejection, chronic
thyroiditis, Graves' disease, schleroderma, diabetes (type I and
type II), active hepatitis (acute and chronic), primary binary
cirrhosis, myasthenia gravis, multiple sclerosis (MS), systemic
lupus erythematosus, atopic dermatitis, contact dermatitis, skin
sunburns, chronic renal insufficiency, Stevens-Johnson syndrome,
idiopathic sprue, sarcoidosis, Guillain-Barre syndrome, uveitis,
conjunctivitis, keratoconjunctivitis, otitis media, periodontal
disease, pulmonary interstitial fibrosis, asthma, bronchitis,
rhinitis, sinusitis, pneumoconiosis, pulmonary insufficiency
syndrome, pulmonary emphysema, pulmonary fibrosis, silicosis,
chronic inflammatory pulmonary disease (e.g. chronic obstructive
pulmonary disease) and other inflammatory or obstructive diseases
of the airways.
[0250] In one embodiment, the HDAC6 inhibitors of the invention are
useful for treating any one or more of the following autoimmune
diseases or disorders, systemic lupus erythematosis, autoimmune
thyroiditis, dermatitis (including atopic dermatitis and eczematous
dermatitis), psoriasis, including keratoconjunctivitis sicca
secondary to Sjogren's Syndrome, alopecia areata, allergic
responses due to arthropod bite reactions, aphthous ulcer, iritis,
conjunctivitis, allergic asthma, cutaneous lupus erythematosus,
scleroderma, vaginitis, proctitis, drug eruptions, leprosy reversal
reactions, erythema nodosum leprosum, autoimmune uveitis, allergic
encephalomyelitis, acute necrotizing hemorrhagic encephalopathy,
idiopathic bilateral progressive sensorineural hearing loss,
idiopathic thrombocytopenia, polychondritis, Wegener's
granulomatosis, chronic active hepatitis, lichen planus,
sarcoidosis, primary biliary cirrhosis, uveitis posterior, and
interstitial lung fibrosis.
[0251] Additionally, the methods of the invention may also be
useful in the treatment of protozoal infections. The methods of the
invention are also useful in the treatment of diseases associated
with aberrant protein catabolism, for example, protein degradation
disorders, disorders associated with misfolded proteins, and
protein deposition disorders. In certain embodiments, the HDAC6
inhibitors of the invention are useful in the treatment of the
protein deposition disorders, Wilson's disease, spinocerebellar
ataxia, prion disease, Parkinson's disease, Huntington's disease,
amyotrophic lateral sclerosis, spinal muscular atrophy, spinal and
bulbar muscular atrophy, amyloidosis, Alzheimer's disease,
Alexander's disease, alcoholic liver disease, cystic fibrosis,
Pick's disease, and Lewy body dementia. In certain exemplary
embodiments, the claimed methods of the invention are useful for
treating disorders associated with histone deacetylation activity.
In certain exemplary embodiments, the methods of the invention are
useful for treating disorders associated with tubulin deacetylation
activity.
[0252] Neurodegenerative diseases that can be treated or prevented
include Alzheimer's disease, Parkinson's disease, cerebral
ischaemia, traumatic neurodegenerative disease, Huntington's
disease or chorea, senile dementia, memory disorder, vascular
dementia, lesions associated with cerebral ischemia (stroke) and
with cranial and medullary trauma, among others.
[0253] Methods delineated herein include those wherein the subject
is identified as in need of a particular stated treatment.
Identifying a subject in need of such treatment can be in the
judgment of a subject or a health care professional and can be
subjective (e.g. opinion) or objective (e.g. measurable by a test
or diagnostic method).
[0254] Preferably, the HDAC6 inhibitors are selective inhibitors of
HDAC6 and, as such, are useful in the treatment of disorders
modulated by histone deacetylases. In one embodiment, the HDAC6
inhibitors of the invention are selective inhibitors of tubulin
deacetylases and, as such, are useful in the treatment of disorders
modulated by tubulin deacetylases.
[0255] Thus, in another aspect of the invention, methods for the
treatment of cancer are provided comprising administering a
therapeutically effective amount of an HDAC6 inhibitor, as
described herein, to a subject in need thereof. In certain
embodiments, the subject is identified as in need of such
treatment. In certain embodiments, a method for the treatment of a
diseases is provided comprising administering a therapeutically
effective amount of an HDAC6 inhibitor, or a pharmaceutical
composition comprising an HDAC6 inhibitor to a subject in need
thereof, in such amounts and for such time as is necessary to
achieve the desired result. In certain embodiments of the present
invention a "therapeutically effective amount" of an HDAC6
inhibitor or pharmaceutical composition is that amount effective
for modulating a subset of a group of cytokines according to the
invention.
[0256] In certain embodiments, the method involves the
administration of a therapeutically effective amount of an HDAC6
inhibitor or a pharmaceutically acceptable derivative thereof to a
subject (including, but not limited to a human or animal) in need
of it (including a subject identified as in need). In certain
embodiments, the HDAC6 inhibitors are useful for the treatment of
cancer (including, but not limited to, glioblastoma,
retinoblastoma, breast cancer, cervical cancer, colon and rectal
cancer, leukemia (e.g., CML, AML, CLL, ALL), lymphoma, lung cancer
(including, but not limited to small cell lung cancer), melanoma
and/or skin cancer, multiple myeloma, non-Hodgkin's lymphoma,
ovarian cancer, pancreatic cancer, prostate cancer and gastric
cancer, bladder cancer, uterine cancer, kidney cancer, testicular
cancer, stomach cancer, brain cancer, liver cancer, or esophageal
cancer, melanoma or multiple melanoma). In certain embodiments, the
HDAC6 inhibitors of the invention are active against leukemia cells
and melanoma cells, and thus are useful for the treatment of
leukemias (e.g., myeloid, lymphocytic, myelocytic and lymphoblastic
leukemias) and malignant melanomas. In still other embodiments, the
inventive anticancer agents are active against solid tumors.
Accordingly, in yet another aspect, according to the methods of
treatment of the present invention, tumor cells are killed, or
their growth is inhibited by contacting said tumor cells with an
HDAC6 inhibitor, as described herein.
[0257] In certain embodiments, the invention provides a method of
treatment of any of the disorders described herein, wherein the
subject is a human.
[0258] In accordance with the foregoing, the present invention
further provides a method for preventing or treating any of the
diseases or disorders described above in a subject in need of such
treatment, which method comprises administering to said subject a
therapeutically effective amount of an HDAC6 inhibitor of the
invention or a pharmaceutically acceptable salt thereof. For any of
the above uses, the required dosage will vary depending on the mode
of administration, the particular condition to be treated and the
effect desired.
[0259] In another aspect, the invention provides a method of
treating a subject suffering from or susceptible to multiple
myeloma comprising administering to a subject in need thereof a
therapeutically effective amount of a compound of formula I:
##STR00182##
[0260] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0261] each of X.sub.1, X.sub.2, X.sub.3, or X.sub.4 is
independently N or CR', wherein 2 of X.sub.1, X.sub.2, X.sub.3, or
X.sub.4 are N;
[0262] each R' is independently H, optionally substituted alkyl,
optionally substituted aryl, or halo;
[0263] R.sub.1 is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; and
[0264] R.sub.2 is H, alkyl, haloalkyl, alkenyl, arylalkyl, aryl,
heteroaryl, heterocyclic, carbocyclic, --C(O)R.sub.5, or
--S(O).sub.pR.sub.5, each of which may be optionally
substituted:
[0265] p is 0, 1, or 2;
[0266] each R.sub.5 is independently H: optionally substituted
alkyl, optionally substituted carbocyclic, optionally substituted
heterocyclic, optionally substituted aryl, or optionally
substituted heteroaryl;
[0267] wherein R.sub.1 and R.sub.2 are both not simultaneously
H;
[0268] to thereby treat the subject suffering from or susceptible
to multiple myeloma.
[0269] In another embodiment, the methods herein comprise a step
wherein a compound is identified as having a Z-fold selectivity for
HDAC6 over other HDACs (e.g., HDAC1, HDAC2, HDAC3), wherein Z is an
integer between 2 and 1000.
[0270] In one embodiment, the invention provides a method further
comprising co-administering one or more of a chemotherapeutic
agent, radiation agent, hormonal agent, biological agent or an
anti-inflammatory agent to the subject.
[0271] In certain embodiments, the chemotherapeutic agent is
tamoxifen, trastuzamab, raloxifene, doxorubicin, fluorouracil/5-fu,
pamidronate disodium, anastrozole, exemestane, cyclophosphamide,
epirubicin, letrozole, toremifene, fulvestrant, fluoxymesterone,
trastuzumab, methotrexate, megastrol acetate, docetaxel,
paclitaxel, testolactone, aziridine, vinblastine, capecitabine,
goselerin acetate, zoledronic acid, taxol, vinblastine, or
vincristine.
[0272] In other embodiments, the invention provides any method as
described herein, wherein the subject is a human.
[0273] In another aspect, the invention provides a kit comprising a
compound capable of inhibiting HDAC activity selected from one or
more compounds of any of the formulae herein (e.g., formula I),
##STR00183##
[0274] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein,
[0275] each of X.sub.1, X.sub.2, X.sub.3, or X.sub.4 is
independently N or CR', wherein 2 of X.sub.1, X.sub.2, X.sub.3, or
X.sub.4 are N;
[0276] each R' is independently H, optionally substituted alkyl,
optionally substituted aryl, or halo;
[0277] R.sub.1 is H, alkyl, haloalkyl, alkenyl, aryl, arylalkyl,
heteroarylalkyl, heteroaryl, heterocyclic, carbocyclic,
--C(O)R.sub.5, or --S(O).sub.pR.sub.5; each of which may be
optionally substituted; and
[0278] R.sub.2 is H, alkyl, haloalkyl, alkenyl, arylalkyl, aryl,
heteroaryl, heterocyclic, carbocyclic, --C(O)R.sub.5, or
--S(O).sub.pR.sub.5, each of which may be optionally
substituted;
[0279] p is 0, 1, or 2;
[0280] each R.sub.5 is independently H; optionally substituted
alkyl, optionally substituted carbocyclic, optionally substituted
heterocyclic, optionally substituted aryl, or optionally
substituted heteroaryl;
wherein R.sub.1 and R.sub.2 are both not simultaneously H;
[0281] and instructions for use in treating multiple myeloma.
[0282] As discussed above, the present invention provides compounds
useful for the treatment of various diseases. In certain
embodiments, the compounds of the present invention are useful as
inhibitors of histone or tubulin deacetylases and thus are useful
as anti-cancer agents, and thus may be useful in the treatment of
cancer, by effecting tumor cell death or inhibiting the growth of
tumor cells. In certain exemplary embodiments, the inventive
anticancer agents are useful in the treatment of cancers and other
proliferative disorders, as described above. In certain
embodiments, the inventive anticancer agents are active against
leukemia cells and melanoma cells, and thus are useful for the
treatment of leukemias (e.g., myeloid, lymphocytic, myelocytic and
lymphoblastic leukemias) and malignant melanomas. In certain
embodiments, the compounds are useful in the treatment of multiple
myeloma.
[0283] The compounds of the invention are also effective to treat
or prevent autoimmune hematologic disorders (e.g. hemolytic anemia,
aplasic anemia, idiopathic thrombocytopenia and neutropenia),
chronic inflammatory pulmonary disease (e.g. chronic obstructive
pulmonary disease) and other inflammatory or obstructive diseases
of the airways.
[0284] Additionally, the inventive compounds may also be useful in
the treatment of protozoal infections. The inventive compounds are
also useful in the treatment of diseases associated with aberrant
protein catabolism, for example, protein degradation disorders,
disorders associated with misfolded proteins, and protein
deposition disorders. In certain embodiments, the compounds are
useful in the treatment of the protein deposition disorders,
Wilson's disease, spinocerebellar ataxia, prion disease,
Parkinson's disease, Huntington's disease, amyotrophic lateral
sclerosis, spinal muscular atrophy, spinal and bulbar muscular
atrophy, amyloidosis, Alzheimer's disease, Alexander's disease,
alcoholic liver disease, cystic fibrosis, Pick's disease, and Lewy
body dementia. In certain exemplary embodiments, the compounds of
the invention are useful for disorders associated with histone
deacetylation activity. In certain exemplary embodiments, the
compounds of the invention are useful for disorders associated with
tubulin deacetylation activity.
[0285] Neurodegenerative diseases that can be treated or prevented
include Alzheimer's disease, Parkinson's disease, cerebral
ischaemia, traumatic neurodegenerative disease, Huntingtons's
disease or chorea, senile dementia, memory disorder, vascular
dementia, lesions associated with cerebral ischemia (stroke) and
with cranial and medullary trauma, among others.
[0286] Methods delineated herein include those wherein the subject
is identified as in need of a particular stated treatment.
Identifying a subject in need of such treatment can be in the
judgment of a subject or a health care professional and can be
subjective (e.g. opinion) or objective (e.g. measurable by a test
or diagnostic method).
[0287] As discussed above, the compounds of the invention are
selective inhibitors of HDAC6 and, as such, are useful in the
treatment of disorders modulated by histone deacetylases. As
discussed above, the compounds of the invention are selective
inhibitors of tubulin deacetylases and, as such, are useful in the
treatment of disorders modulated by tubulin deacetylases. For
example, compounds of the invention may be useful in the treatment
of cancer (e.g., breast cancer, prostate cancer, multiple myeloma,
leukemia, lymphoma, etc.). Accordingly, in yet another aspect,
according to the methods of treatment of the present invention,
tumor cells are killed, or their growth is inhibited by contacting
said tumor cells with an inventive compound or composition, as
described herein.
[0288] Thus, in another aspect of the invention, methods for the
treatment of cancer are provided comprising administering a
therapeutically effective amount of an inventive compound (i.e., of
any of the formulae herein), as described herein, to a subject in
need thereof. In certain embodiments, the subject is identified as
in need of such treatment. In certain embodiments, a method for the
treatment of cancer is provided comprising administering a
therapeutically effective amount of an inventive compound, or a
pharmaceutical composition comprising an inventive compound to a
subject in need thereof, in such amounts and for such time as is
necessary to achieve the desired result. In certain embodiments of
the present invention a "therapeutically effective amount" of the
inventive compound or pharmaceutical composition is that amount
effective for killing or inhibiting the growth of tumor cells. The
compounds and compositions, according to the method of the present
invention, may be administered using any amount and any route of
administration effective for killing or inhibiting the growth of
tumor cells. Thus, the expression "amount effective to kill or
inhibit the growth of tumor cells," as used herein, refers to a
sufficient amount of agent to kill or inhibit the growth of tumor
cells. The exact amount required will vary from subject to subject,
depending on the species, age, and general condition of the
subject, the severity of the infection, the particular anticancer
agent, its mode of administration, and the like.
[0289] In certain embodiments, the method involves the
administration of a therapeutically effective amount of the
compound or a pharmaceutically acceptable derivative thereof to a
subject (including, but not limited to a human or animal) in need
of it.
[0290] Multiple myeloma (MM) is a plasma cell malignancy which
remains incurable despite conventional treatment (Gregory, et al.
(1992) J Clin Oncol 10, 334-342) as well as high dose therapy and
stem cell transplantation (Attal, M., et al. (2003) N Engl J Med
349, 2495-2502). Novel agents have recently been developed which
target not only MM cells, but also the bone marrow (BM)
microenvironment, and can overcome conventional drug resistance
(Hideshima, T. & Anderson, K. C. (2002) Nat Rev Cancer 2,
927-937). For example, the proteasome inhibitor bortezomib
(formally PS-341) induces significant anti-tumor activity in human
MM cell lines and freshly isolated patient MM cells (Hideshima, T.
& Anderson, K. C. (2002) Nat Rev Cancer 2, 927-937; Hideshima,
et al. (2001) Cancer Res. 61, 3071-3076; Mitsiades, N., et al.
(2002) Proc Natl Acad Sci USA 99, 14374-14379; Hideshima, T., et
al. (2002) J Biol Chem 277, 16639-47: Mitsiades, N., et al. (2003)
Blood 101, 2377-80; Chauhan, D., et al (2003) Cancer Res 63,
6174-6177; Hideshima, T., et al. (2003) Blood 101, 1530-1534;
Hideshima, T., et al. (2003) Oncogene 22, 8386-8393; Hideshima, T.,
et al. (2004) Oncogene 23, 8766-8776) associated with c-Jun
NH2-terminal kinase (JNK) (also known as stress-activated protein
kinase) and caspase activation, followed by apoptosis (Hideshima,
T., et al. (2001) Cancer Res. 61, 3071-3076; Mitsiades, N., et al.
(2002) Proc Natl Acad Sci USA 99, 14374-14379; Hideshima, T., et
al. (2003) Blood 101, 1530-1534). Bortezomib also inhibits
adherence of MM cells to bone marrow stromal cells (BMSCs) by
downregulating adhesion molecules (ICAM-I and VCAM-I) (Hideshima,
T., et al. (2001) Oncogene 20, 4519-4527); as well as induces
cleavage of DNA-protein kinase catalytic subunit and ataxia
telangiectasia mutated, suggesting that bortezomib also inhibits
DNA repair. Neither IL-6 nor adherence of MM cells to BMSCs
protects against bortezomib-induced apoptosis. Without wishing to
be bound by any scientific theory, bortezomib enhances sensitivity
and can overcome resistance in MM cells to conventional
chemotherapeutic agents, especially to DNA damaging agents
(Mitsiades, N., et al. (2003) Blood 101, 2377-80). In support of
this, a phase II trial of bortezomib treatment of 202 patients with
refractory relapsed MM demonstrated 35% responses, including 10%
complete and near complete responses (Richardson, P. G., et al.
(2003) NEnglJMed 348, 2609-2617); however, 65% of patients did not
respond. Heat-shock protein (hsp)-27 mediates
bortezomib-resistance; conversely, inhibiting hsp-27 expression
using hsp-27 antisense, p38 mitogen-activated protein kinase (MAPK)
siRNA, or p38 MAPK inhibitor to downregulate hsp-27 can restore MM
cell susceptibility to bortezomib (Chauhan, D., et al. (2003)
Cancer Res 63, 6174-6177; Hideshima, T., et al. (2004) Oncogene 23,
8766-8776).
[0291] In certain embodiments, the inventive compounds also find
use in the prevention of restenosis of blood vessels subject to
traumas such as angioplasty and stenting. For example, it is
contemplated that the compounds of the invention will be useful as
a coating for implanted medical devices, such as tubings, shunts,
catheters, artificial implants, pins, electrical implants such as
pacemakers, and especially for arterial or venous stents, including
balloon-expandable stents. In certain embodiments inventive
compounds may be bound to an implantable medical device, or
alternatively, may be passively adsorbed to the surface of the
implantable device. In certain other embodiments, the inventive
compounds may be formulated to be contained within, or, adapted to
release by a surgical or medical device or implant, such as, for
example, stents, sutures, indwelling catheters, prosthesis, and the
like. Accordingly, without wishing to be bound to any particular
theory, the inventive compounds having antiproliferative effects
can be used as stent coatings and/or in stent drug delivery
devices, inter alia for the prevention of restenosis or reduction
of restenosis rate. Suitable coatings and the general preparation
of coated implantable devices are described in U.S. Pat. Nos.
6,099,562; 5,886,026; and 5,304,121; each of which is incorporated
herein by reference. The coatings are typically biocompatible
polymeric materials such as a hydrogel polymer,
polymethyldisiloxane. polycaprolactone, polyethylene glycol,
polylactic acid, ethylene vinyl acetate, and mixtures thereof. The
coatings may optionally be further covered by a suitable topcoat of
fluorosilicone, polysaccarides, polyethylene glycol, phospholipids
or combinations thereof to impart controlled release
characteristics in the composition. A variety of compositions and
methods related to stent coating and/or local stent drug delivery
for preventing restenosis are known in the art (see, for example,
U.S. Pat. Nos. 6,517,889; 6,273,913; 6,258,121; 6,251,136;
6,248,127; 6,231,600; 6,203,551; 6,153,252; 6,071,305; 5,891,507;
5,837,313 and published U.S. patent application No.:
US2001/0027340, each of which is incorporated herein by reference
in its entirety). For example, stents may be coated with
polymer-drug conjugates by dipping the stent in polymer-drug
solution or spraying the stent with such a solution. In certain
embodiments, suitable materials for the implantable device include
biocompatible and nontoxic materials, and may be chosen from the
metals such as nickel-titanium alloys, steel, or biocompatible
polymers, hydrogels, polyurethanes, polyethylenes, ethylenevinyl
acetate copolymers, etc. In certain embodiments, the inventive
compound is coated onto a stent for insertion into an artery or
vein following balloon angioplasty.
[0292] The compounds of this invention or pharmaceutically
acceptable compositions thereof may also be incorporated into
compositions for coating implantable medical devices, such as
prostheses, artificial valves, vascular grafts, stents and
catheters. Accordingly, the present invention, in another aspect,
includes a composition for coating an implantable device comprising
a compound of the present invention as described generally above,
and a carrier suitable for coating said implantable device. In
still another aspect, the present invention includes an implantable
device coated with a composition comprising a compound of the
present invention as described generally above, and a carrier
suitable for coating said implantable device.
[0293] Within other aspects of the present invention, methods are
provided for expanding the lumen of a body passageway, comprising
inserting a stent into the passageway, the stent having a generally
tubular structure, the surface of the structure being coated with
(or otherwise adapted to release) an inventive compound or
composition, such that the passageway is expanded. In certain
embodiments, the lumen of a body passageway is expanded in order to
eliminate a biliary, gastrointestinal, esophageal,
tracheal/bronchial, urethral and/or vascular obstruction.
[0294] In certain embodiments, the invention provides a method of
treatment of any of the disorders described herein, wherein the
subject is a human.
[0295] In accordance with the foregoing, the present invention
further provides a method for preventing or treating any of the
diseases or disorders described above in a subject in need of such
treatment, which method comprises administering to said subject a
therapeutically effective amount of a compound of the invention or
a pharmaceutically acceptable salt thereof. For any of the above
uses, the required dosage will vary depending on the mode of
administration, the particular condition to be treated and the
effect desired.
[0296] Pharmaceutical Compositions
[0297] In another aspect, the invention provides a pharmaceutical
composition comprising a compound of any of the formulae herein
(e.g., formula I), or a pharmaceutically acceptable ester, salt, or
prodrug thereof, together with a pharmaceutically acceptable
carrier.
[0298] Compounds of the invention can be administered as
pharmaceutical compositions by any conventional route, in
particular enterally, e.g., orally, e.g., in the form of tablets or
capsules, or parenterally, e.g., in the form of injectable
solutions or suspensions, topically, e.g., in the form of lotions,
gels, ointments or creams, or in a nasal or suppository form.
Pharmaceutical compositions comprising a compound of the present
invention in free form or in a pharmaceutically acceptable salt
form in association with at least one pharmaceutically acceptable
carrier or diluent can be manufactured in a conventional manner by
mixing, granulating or coating methods. For example, oral
compositions can be tablets or gelatin capsules comprising the
active ingredient together with a) diluents, e.g., lactose,
dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine; b)
lubricants, e.g., silica, talcum, stearic acid, its magnesium or
calcium salt and/or polyethyleneglycol: for tablets also c)
binders, e.g., magnesium aluminum silicate, starch paste, gelatin,
tragacanth, methylcellulose, sodium carboxymethylcellulose and or
polyvinylpyrrolidone: if desired d) disintegrants, e.g., starches,
agar, alginic acid or its sodium salt, or effervescent mixtures;
and/or e) absorbents, colorants, flavors and sweeteners. Injectable
compositions can be aqueous isotonic solutions or suspensions, and
suppositories can be prepared from fatty emulsions or suspensions.
The compositions may be sterilized and/or contain adjuvants, such
as preserving, stabilizing, wetting or emulsifying agents, solution
promoters, salts for regulating the osmotic pressure and/or
buffers. In addition, they may also contain other therapeutically
valuable substances. Suitable formulations for transdermal
applications include an effective amount of a compound of the
present invention with a carrier. A carrier can include absorbable
pharmacologically acceptable solvents to assist passage through the
skin of the host. For example, transdermal devices are in the form
of a bandage comprising a backing member, a reservoir containing
the compound optionally with carriers, optionally a rate
controlling barrier to deliver the compound to the skin of the host
at a controlled and predetermined rate over a prolonged period of
time, and means to secure the device to the skin. Matrix
transdermal formulations may also be used. Suitable formulations
for topical application, e.g., to the skin and eyes, are preferably
aqueous solutions, ointments, creams or gels well-known in the art.
Such may contain solubilizers, stabilizers, tonicity enhancing
agents, buffers and preservatives.
[0299] Compounds of the invention can be administered in
therapeutically effective amounts in combination with one or more
therapeutic agents (pharmaceutical combinations). For example,
synergistic effects can occur with other anti-proliferative,
anti-cancer, immunomodulatory or anti-inflammatory substances.
Where the compounds of the invention are administered in
conjunction with other therapies, dosages of the co-administered
compounds will of course vary depending on the type of co-drug
employed, on the specific drug employed, on the condition being
treated and so forth.
[0300] Combination therapy includes the administration of the
subject compounds in further combination with other biologically
active ingredients (such as, but not limited to, a second and
different antineoplastic agent) and non-drug therapies (such as,
but not limited to, surgery or radiation treatment). For instance,
the compounds of the invention can be used in combination with
other pharmaceutically active compounds, preferably compounds that
are able to enhance the effect of the compounds of the invention.
The compounds of the invention can be administered simultaneously
(as a single preparation or separate preparation) or sequentially
to the other drug therapy. In general, a combination therapy
envisions administration of two or more drugs during a single cycle
or course of therapy.
[0301] In certain embodiments, these compositions optionally
further comprise one or more additional therapeutic agents.
Alternatively, a compound of this invention may be administered to
a patient in need thereof in combination with the administration of
one or more other therapeutic agents. For example, additional
therapeutic agents for conjoint administration or inclusion in a
pharmaceutical composition with a compound of this invention may be
an approved chemotherapeutic agent, or it may be any one of a
number of agents undergoing approval in the Food and Drug
Administration that ultimately obtain approval for the treatment of
protozoal infections and/or any disorder associated with cellular
hyperproliferation. In certain other embodiments, the additional
therapeutic agent is an anticancer agent, as discussed in more
detail herein. In certain other embodiments, the compositions of
the invention are useful for the treatment of protozoal infections.
In the treatment of cancer or protein degradation disorders, the
inventive compound may be combined with a proteasome inhibitor
(e.g., bortezomib, Rl 15777 FTI, MG 132, NPI-0052, etc.). In the
treatment of cancer or protein degradation disorders, the inventive
compound may be combined with protein degradation inhibitor (e.g.
another inventive compound, a tubacin-like compound, bortezomib, Rl
15777 FTI, MGl 32, NPI-0052, SAHA, .sup.166Ho-DOTMP, arsenic
trioxide, 17 AAG, MG 132, etc.).
[0302] It will also be appreciated that the compounds and
pharmaceutical compositions of the present invention can be
employed in combination therapies, that is, the compounds and
pharmaceutical compositions can be administered concurrently with,
prior to, or subsequent to, one or more other desired therapeutics
or medical procedures. The particular combination of therapies
(therapeutics or procedures) to employ in a combination regimen
will take into account compatibility of the desired therapeutics
and/or procedures and the desired therapeutic effect to be
achieved. It will also be appreciated that the therapies employed
may achieve a desired effect for the same disorder (for example, an
inventive compound may be administered concurrently with another
anticancer agent), or they may achieve different effects (e.g.,
control of any adverse effects).
[0303] The present invention encompasses pharmaceutically
acceptable topical formulations of inventive compounds. The term
"pharmaceutically acceptable topical formulation," as used herein,
means any formulation which is pharmaceutically acceptable for
intradermal administration of a compound of the invention by
application of the formulation to the epidermis. In certain
embodiments of the invention, the topical formulation comprises a
carrier system. Pharmaceutically effective carriers include, but
are not limited to, solvents (e.g., alcohols, poly alcohols,
water), creams, lotions, ointments, oils, plasters, liposomes,
powders, emulsions, microemulsions, and buffered solutions (e.g.,
hypotonic or buffered saline) or any other carrier known in the art
for topically administering pharmaceuticals. A more complete
listing of art-known carriers is provided by reference texts that
are standard in the art, for example, Remington's Pharmaceutical
Sciences, 16th Edition, 1980 and 17th Edition, 1985, both published
by Mack Publishing Company, Easton, Pa., the disclosures of which
are incorporated herein by reference in their entireties. In
certain other embodiments, the topical formulations of the
invention may comprise excipients. Any pharmaceutically acceptable
excipient known in the art may be used to prepare the inventive
pharmaceutically acceptable topical formulations. Examples of
excipients that can be included in the topical formulations of the
invention include, but are not limited to, preservatives,
antioxidants, moisturizers, emollients, buffering agents,
solubilizing agents, other penetration agents, skin protectants,
surfactants, and propellants, and/or additional therapeutic agents
used in combination to the inventive compound. Suitable
preservatives include, but are not limited to, alcohols, quaternary
amines, organic acids, parabens, and phenols. Suitable antioxidants
include, but are not limited to, ascorbic acid and its esters,
sodium bisulfite, butylated hydroxytoluene, butylated
hydroxyanisole, tocopherols, and chelating agents like EDTA and
citric acid. Suitable moisturizers include, but are not limited to,
glycerine, sorbitol, polyethylene glycols, urea, and propylene
glycol. Suitable buffering agents for use with the invention
include, but are not limited to, citric, hydrochloric, and lactic
acid buffers. Suitable solubilizing agents include, but are not
limited to, quaternary ammonium chlorides, cyclodextrins, benzyl
benzoate, lecithin, and polysorbates. Suitable skin protectants
that can be used in the topical formulations of the invention
include, but are not limited to, vitamin E oil, allatoin,
dimethicone, glycerin, petrolatum, and zinc oxide.
[0304] In certain embodiments, the pharmaceutically acceptable
topical formulations of the invention comprise at least a compound
of the invention and a penetration enhancing agent. The choice of
topical formulation will depend or several factors, including the
condition to be treated, the physicochemical characteristics of the
inventive compound and other excipients present, their stability in
the formulation, available manufacturing equipment, and costs
constraints. As used herein the term "penetration enhancing agent"
means an agent capable of transporting a pharmacologically active
compound through the stratum corneum and into the epidermis or
dermis, preferably, with little or no systemic absorption. A wide
variety of compounds have been evaluated as to their effectiveness
in enhancing the rate of penetration of drugs through the skin.
See, for example, Percutaneous Penetration Enhancers, Maibach H. I.
and Smith H. E. (eds.), CRC Press, Inc., Boca Raton, Fla. (1995),
which surveys the use and testing of various skin penetration
enhancers, and Buyuktimkin et al., Chemical Means of Transdermal
Drug Permeation Enhancement in Transdermal and Topical Drug
Delivery Systems, Gosh T. K., Pfister W. R., Yum S. I. (Eds.),
Interpharm Press Inc., Buffalo Grove, Ill. (1997). In certain
exemplary embodiments, penetration agents for use with the
invention include, but are not limited to, triglycerides (e.g.,
soybean oil), aloe compositions (e.g., aloe-vera gel), ethyl
alcohol, isopropyl alcohol, octolyphenylpolyethylene glycol, oleic
acid, polyethylene glycol 400, propylene glycol,
N-decylmethylsulfoxide, fatty acid esters (e.g., isopropyl
myristate, methyl laurate, glycerol monooleate, and propylene
glycol monooleate) and N-methyl pyrrolidine.
[0305] In certain embodiments, the compositions may be in the form
of ointments, pastes, creams, lotions, gels, powders, solutions,
sprays, inhalants or patches. In certain exemplary embodiments,
formulations of the compositions according to the invention are
creams, which may further contain saturated or unsaturated fatty
acids such as stearic acid, palmitic acid, oleic acid,
palmito-oleic acid, cetyl or oleyl alcohols, stearic acid being
particularly preferred. Creams of the invention may also contain a
non-ionic surfactant, for example, polyoxy-40-stearate. In certain
embodiments, the active component is admixed under sterile
conditions with a pharmaceutically acceptable carrier and any
needed preservatives or buffers as may be required. Ophthalmic
formulation, eardrops, and eye drops are also contemplated as being
within the scope of this invention. Additionally, the present
invention contemplates the use of transdermal patches, which have
the added advantage of providing controlled delivery of a compound
to the body. Such dosage forms are made by dissolving or dispensing
the compound in the proper medium. As discussed above, penetration
enhancing agents can also be used to increase the flux of the
compound across the skin. The rate can be controlled by either
providing a rate controlling membrane or by dispersing the compound
in a polymer matrix or gel.
[0306] It will also be appreciated that the compounds and
pharmaceutical compositions of the present invention can be
formulated and employed in combination therapies, that is, the
compounds and pharmaceutical compositions can be formulated with or
administered concurrently with, prior to, or subsequent to, one or
more other desired therapeutics or medical procedures. The
particular combination of therapies (therapeutics or procedures) to
employ in a combination regimen will take into account
compatibility of the desired therapeutics and/or procedures and the
desired therapeutic effect to be achieved. It will also be
appreciated that the therapies employed may achieve a desired
effect for the same disorder (for example, an inventive compound
may be administered concurrently with another immunomodulatory
agent, anticancer agent or agent useful for the treatment of
psoriasis), or they may achieve different effects (e.g., control of
any adverse effects).
[0307] For example, other therapies or anticancer agents that may
be used in combination with the inventive compounds of the present
invention include, but not limited to, surgery, radiotherapy (in
but a few examples, gamma-radiation, neutron beam radiotherapy,
electron beam radiotherapy, proton therapy, brachytherapy, and
systemic radioactive isotopes, to name a few), endocrine therapy,
biologic response modifiers (interferons, interleukins, antibodies,
aptamers, siRNAs, oligonucletoides, enzyme, ion channel and
receptor inhibitors or activators to name a-few), hyperthermia and
cryotherapy, agents to attenuate any adverse effects (e.g.,
antiemetics), and other approved chemotherapeutic drugs, including,
but not limited to, alkylating drugs (e.g., mechlorethamine,
chlorambucil, Cyclophosphamide, Melphalan, Ifosfamide),
antimetabolites (e.g., Methotrexate), purine antagonists and
pyrimidine antagonists (e.g., 6-Mercaptopurine, 5-Fluorouracil,
Cytarabile, Gemcitabine), spindle poisons (e.g., Vinblastine,
Vincristine, Vinorelbine, Paclitaxel), podophyllotoxins (e.g.,
Etoposide, Irinotecan, Topotecan), antibiotics (Doxorubicin,
Bleomycin, Mitomycin), nitrosoureas (e.g., Carmustine, Lomustine),
inorganic ions (e.g., Cisplatin, Carboplatin), enzymes (e.g.,
Asparaginase), and hormones (e.g., Tamoxifen, Leuprolide,
Flutamide, and Megestrol), to name a few. For a more comprehensive
discussion of updated cancer therapies see, The Merck Manual,
Seventeenth Ed. 1999, the entire contents of which are hereby
incorporated by reference. See also the National Cancer Institute
(NCI) website (www.nci.nih.gov) and the Food and Drug
Administration (FDA) website for a list of the FDA approved
oncology drugs (www.fda.gov/cder/cancer/dmglistfrarne).
[0308] In certain embodiments, the pharmaceutical compositions of
the present invention further comprise one or more additional
therapeutically active ingredients (e.g., chemotherapeutic and/or
palliative). For purposes of the invention, the term "palliative"
refers to treatment that is focused on the relief of symptoms of a
disease and/or side effects of a therapeutic regimen, but is not
curative. For example, palliative treatment encompasses
painkillers, antinausea medications, anti-pyretics, and
anti-sickness drugs. In addition, chemotherapy, radiotherapy and
surgery can all be used palliatively (that is, to reduce symptoms
without going for cure; e.g., for shrinking tumors and reducing
pressure, bleeding, pain and other symptoms of cancer).
[0309] The present compounds and compositions can be administered
together with hormonal and steroidal anti-inflammatory agents, such
as but not limited to, estradiol, conjugated estrogens (e.g.,
PREMARIN, PREMPRO, AND PREMPHASE), 17 beta estradiol,
calcitonin-salmon, levothyroxine, dexamethasone,
medroxyprogesterone, prednisone, cortisone, flunisolide, and
hydrocortisone; non-steroidal anti-inflammatory agents, such as but
not limited to, tramadol, fentanyl, metamizole, ketoprofen,
naproxen, nabumetone, ketoralac, tromethamine, loxoprofen,
ibuprofen, aspirin, and acetaminophen; anti-TNF-.alpha. antibodies,
such as infliximab (REMICADE.RTM.) and etanercept
(ENBREL.RTM.).
[0310] The pharmaceutical compositions of the present invention
comprise a therapeutically effective amount of a compound of the
present invention formulated together with one or more
pharmaceutically acceptable carriers. As used herein, the term
"pharmaceutically acceptable carrier" means a non-toxic, inert
solid, semi-solid or liquid filler, diluent, encapsulating material
or formulation auxiliary of any type. The pharmaceutical
compositions of this invention can be administered to humans and
other animals orally, rectally, parenterally, intracisternally,
intravaginally, intraperitoneally, topically (as by powders,
ointments, or drops), buccally, or as an oral or nasal spray.
[0311] Liquid dosage forms for oral administration include
pharmaceutically acceptable emulsions, microemulsions, solutions,
suspensions, syrups and elixirs. In addition to the active
compounds, the liquid dosage forms may contain inert diluents
commonly used in the art such as, for example, water or other
solvents, solubilizing agents and emulsifiers such as ethyl
alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl
alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol,
dimethylformamide, oils (in particular, cottonseed, groundnut,
corn, germ, olive, castor, and sesame oils), glycerol,
tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid
esters of sorbitan, and mixtures thereof. Besides inert diluents,
the oral compositions can also include adjuvants such as wetting
agents, emulsifying and suspending agents, sweetening, flavoring,
and perfuming agents.
[0312] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution, suspension or emulsion in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, U.S.P.
and isotonic sodium chloride solution. In addition, sterile, fixed
oils are conventionally employed as a solvent or suspending medium.
For this purpose any bland fixed oil can be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid are used in the preparation of injectables.
[0313] In order to prolong the effect of a drug, it is often
desirable to slow the absorption of the drug from subcutaneous or
intramuscular injection. This may be accomplished by the use of a
liquid suspension of crystalline or amorphous material with poor
water solubility. The rate of absorption of the drug then depends
upon its rate of dissolution which, in turn, may depend upon
crystal size and crystalline form. Alternatively, delayed
absorption of a parenterally administered drug form is accomplished
by dissolving or suspending the drug in an oil vehicle.
[0314] Compositions for rectal or vaginal administration are
preferably suppositories which can be prepared by mixing the
compounds of this invention with suitable non-irritating excipients
or carriers such as cocoa butter, polyethylene glycol or a
suppository wax which are solid at ambient temperature but liquid
at body temperature and therefore melt in the rectum or vaginal
cavity and release the active compound.
[0315] Solid compositions of a similar type may also be employed as
fillers in soft and hard-filled gelatin capsules using such
excipients as lactose or milk sugar as well as high molecular
weight polyethylene glycols and the like.
[0316] The active compounds can also be in micro-encapsulated form
with one or more excipients as noted above. The solid dosage forms
of tablets, dragees, capsules, pills, and granules can be prepared
with coatings and shells such as enteric coatings, release
controlling coatings and other coatings well known in the
pharmaceutical formulating art. In such solid dosage forms the
active compound may be admixed with at least one inert diluent such
as sucrose, lactose or starch. Such dosage forms may also comprise,
as is normal practice, additional substances other than inert
diluents, e.g., tableting lubricants and other tableting aids such
a magnesium stearate and microcrystalline cellulose. In the case of
capsules, tablets and pills, the dosage forms may also comprise
buffering agents.
[0317] Dosage forms for topical or transdermal administration of a
compound of this invention include ointments, pastes, creams,
lotions, gels, powders, solutions, sprays, inhalants or patches.
The active component is admixed under sterile conditions with a
pharmaceutically acceptable carrier and any needed preservatives or
buffers as may be required. Ophthalmic formulation, ear drops, eye
ointments, powders and solutions are also contemplated as being
within the scope of this invention.
[0318] The ointments, pastes, creams and gels may contain, in
addition to an active compound of this invention, excipients such
as animal and vegetable fats, oils, waxes, paraffins, starch,
tragacanth, cellulose derivatives, polyethylene glycols, silicones,
bentonites, silicic acid, talc and zinc oxide, or mixtures
thereof.
[0319] Powders and sprays can contain, in addition to the compounds
of this invention, excipients such as lactose, talc, silicic acid,
aluminum hydroxide, calcium silicates and polyamide powder, or
mixtures of these substances. Sprays can additionally contain
customary propellants such as chlorofluorohydrocarbons.
[0320] Transdermal patches have the added advantage of providing
controlled delivery of a compound to the body. Such dosage forms
can be made by dissolving or dispensing the compound in the proper
medium. Absorption enhancers can also be used to increase the flux
of the compound across the skin. The rate can be controlled by
either providing a rate controlling membrane or by dispersing the
compound in a polymer matrix or gel.
[0321] According to the methods of treatment of the present
invention, disorders are treated or prevented in a subject, such as
a human or other animal, by administering to the subject a
therapeutically effective amount of a compound of the invention, in
such amounts and for such time as is necessary to achieve the
desired result. The term "therapeutically effective amount" of a
compound of the invention, as used herein, means a sufficient
amount of the compound so as to decrease the symptoms of a disorder
in a subject. As is well understood in the medical arts a
therapeutically effective amount of a compound of this invention
will be at a reasonable benefit/risk ratio applicable to any
medical treatment.
[0322] In general, compounds of the invention will be administered
in therapeutically effective amounts via any of the usual and
acceptable modes known in the art, either singly or in combination
with one or more therapeutic agents. A therapeutically effective
amount may vary widely depending on the severity of the disease,
the age and relative health of the subject, the potency of the
compound used and other factors. In general, satisfactory results
are indicated to be obtained systemically at daily dosages of from
about 0.03 to 2.5 mg/kg per body weight (0.05 to 4.5 mg/m.sup.2).
An indicated daily dosage in the larger mammal, e.g. humans, is in
the range from about 0.5 mg to about 100 mg, conveniently
administered, e.g. in divided doses up to four times a day or in
controlled release form. Suitable unit dosage forms for oral
administration comprise from ca. 1 to 50 mg active ingredient.
[0323] In certain embodiments, a therapeutic amount or dose of the
compounds of the present invention may range from about 0.1 mg/kg
to about 500 mg/kg (about 0.18 mg/m.sup.2 to about 900 mg/m.sup.2),
alternatively from about 1 to about 50 mg/kg (about 1.8 to about 90
mg/m.sup.2). In general, treatment regimens according to the
present invention comprise administration to a patient in need of
such treatment from about 10 mg to about 1000 mg of the compound(s)
of this invention per day in single or multiple doses. Therapeutic
amounts or doses will also vary depending on route of
administration, as well as the possibility of co-usage with other
agents.
[0324] Upon improvement of a subject's condition, a maintenance
dose of a compound, composition or combination of this invention
may be administered, if necessary. Subsequently, the dosage or
frequency of administration, or both, may be reduced, as a function
of the symptoms, to a level at which the improved condition is
retained when the symptoms have been alleviated to the desired
level, treatment should cease. The subject may, however, require
intermittent treatment on a long-term basis upon any recurrence of
disease symptoms.
[0325] It will be understood, however, that the total daily usage
of the compounds and compositions of the present invention will be
decided by the attending physician within the scope of sound
medical judgment. The specific inhibitory dose for any particular
patient will depend upon a variety of factors including the
disorder being treated and the severity of the disorder, the
activity of the specific compound employed; the specific
composition employed; the age, body weight, general health, sex and
diet of the patient; the time of administration, route of
administration, and rate of excretion of the specific compound
employed: the duration of the treatment; drugs used in combination
or coincidental with the specific compound employed; and like
factors well known in the medical arts.
[0326] The invention also provides for a pharmaceutical
combinations, e.g. a kit, comprising a) a first agent which is a
compound of the invention as disclosed herein, in free form or in
pharmaceutically acceptable salt form, and b) at least one
co-agent. The kit can comprise instructions for its administration
to a subject suffering from or susceptible to a disease or
disorder.
[0327] The terms "co-administration" or "combined administration"
or the like as utilized herein are meant to encompass
administration of the selected therapeutic agents to a single
patient, and are intended to include treatment regimens in which
the agents are not necessarily administered by the same route of
administration or at the same time.
[0328] The term "pharmaceutical combination" as used herein means a
product that results from the mixing or combining of more than one
active ingredient and includes both fixed and non-fixed
combinations of the active ingredients. The term "fixed
combination" means that the active ingredients, e.g., a compound of
the invention and a co-agent, are both administered to a patient
simultaneously in the form of a single entity or dosage. The term
"non-fixed combination" means that the active ingredients, e.g., a
compound of the invention and a co-agent, are both administered to
a patient as separate entities either simultaneously, concurrently
or sequentially with no specific time limits, wherein such
administration provides therapeutically effective levels of the two
compounds in the body of the patient. The latter also applies to
cocktail therapy, e.g., the administration of three or more active
ingredients.
[0329] Some examples of materials which can serve as
pharmaceutically acceptable carriers include, but are not limited
to, ion exchangers, alumina, aluminum stearate, lecithin, serum
proteins, such as human serum albumin, buffer substances such as
phosphates, glycine, sorbic acid, or potassium sorbate, partial
glyceride mixtures of saturated vegetable fatty acids, water, salts
or electrolytes, such as protamine sulfate, disodium hydrogen
phosphate, potassium hydrogen phosphate, sodium chloride, zinc
salts, colloidal silica, magnesium trisilicate, polyvinyl
pyrrolidone, polyacrylates, waxes,
polyethylene-polyoxypropylene-block polymers, wool fat, sugars such
as lactose, glucose and sucrose; starches such as corn starch and
potato starch; cellulose and its derivatives such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients such as cocoa
butter and suppository waxes, oils such as peanut oil, cottonseed
oil; safflower oil; sesame oil; olive oil; corn oil and soybean
oil; glycols; such a propylene glycol or polyethylene glycol;
esters such as ethyl oleate and ethyl laurate, agar; buffering
agents such as magnesium hydroxide and aluminum hydroxide; alginic
acid; pyrogen-free water, isotonic saline; Ringer's solution; ethyl
alcohol, and phosphate buffer solutions, as well as other non-toxic
compatible lubricants such as sodium lauryl sulfate and magnesium
stearate, as well as coloring agents, releasing agents, coating
agents, sweetening, flavoring and perfuming agents, preservatives
and antioxidants can also be present in the composition, according
to the judgment of the formulator. The protein kinase inhibitors or
pharmaceutical salts thereof may be formulated into pharmaceutical
compositions for administration to animals or humans. These
pharmaceutical compositions, which comprise an amount of the
protein inhibitor effective to treat or prevent a protein
kinase-mediated condition and a pharmaceutically acceptable
carrier, are another embodiment of the present invention.
Examples
[0330] The compounds and processes of the present invention will be
better understood in connection with the following examples, which
are intended as an illustration only and not to limit the scope of
the invention. Various changes and modifications to the disclosed
embodiments will be apparent to those skilled in the art and such
changes and modifications including, without limitation, those
relating to the chemical structures, substituents, derivatives,
formulations and/or methods of the invention may be made without
departing from the spirit of the invention and the scope of the
appended claims. Definitions of variables in the structures in
schemes herein are commensurate with those of corresponding
positions in the formulae delineated herein.
Example 1: Synthesis of 1
##STR00184##
##STR00185##
[0331] Synthesis of ethyl
2-(phenylamino)pyrimidine-5-carboxylate
[0332] To a solution of compound A in NMP (20 ml) was added aniline
(1.0 g, 5.36 mmol) and heated at 120.degree. C. for overnight.
After the reaction completed, the mixture was poured into ice water
(20 ml), the precipitate was collected and washed with methanol (5
ml*2), the precipitation was isolated by filtration to afford the
compound B as a pale solid (980 mg, 75%).
##STR00186##
Synthesis of N-hydroxy-2-(phenylamino)pyrimidine-5-carboxamide
[0333] To a solution of compound B (100 mg, 0.411 mmol) in
methanol/dichloromethane (5/2 ml) was added 98% NH.sub.2OH (1.7 ml,
50% in water) at 0.degree. C. and stirred for 10 min. A solution of
saturated sodium hydroxide in methanol (1.5 ml) was added and the
mixture kept stirring for another 10 min. The reaction mixture was
concentrated in vacuum and the crude residue was acidified to
pH=6-7 with 2N HCl. The precipitate was collected, washed with
water to afford 1 as an off-white solid (90 mg, 94.7%).
Example 2: Synthesis of 99
##STR00187##
[0334] Example 3: Synthesis of 110
##STR00188##
[0335] Example 4: Synthesis of 111
##STR00189##
[0336] Example 5: HDAC Enzyme Assays
[0337] Compounds for testing were diluted in DMSO to 50 fold the
final concentration and a ten point three fold dilution series was
made. The compounds were diluted in assay buffer (50 mM HEPES, pH
7.4, 100 mM KCl, 0.001% Tween-20, 0.05% BSA, 20 .mu.M TCEP) to 6
fold their final concentration. The HDAC enzymes (purchased from
BPS Biosciences) were diluted to 1.5 fold their final concentration
in assay buffer. The tripeptide substrate and trypsin at 0.05 .mu.M
final concentration were diluted in assay buffer at 6 fold their
final concentration. The final enzyme concentrations used in these
assays were 3.3 ng/ml (HDAC1), 0.2 ng/ml (HDAC2), 0.08 ng/ml
(HDAC3) and 2 ng/ml (HDAC6). The final substrate concentrations
used were 16 .mu.M (HDAC1), 10 .mu.M (HDAC2), 17 .mu.M (HDAC3) and
14 .mu.M (HDAC6). Five .mu.l of compounds and 20 .mu.l of enzyme
were added to wells of a black, opaque 384 well plate in duplicate.
Enzyme and compound were incubated together at room temperature for
10 minutes. Five .mu.l of substrate was added to each well, the
plate was shaken for 60 seconds and placed into a Victor 2
microtiter plate reader. The development of fluorescence was
monitored for 60 min and the linear rate of the reaction was
calculated. The IC.sub.50 was determined using Graph Pad Prism by a
four parameter curve fit.
TABLE-US-00003 IC.sub.50 (nM) Compound ID HDAC1 HDAC2 HDAC3 HDAC6 2
264 (53.1x) 248 (50x) 299 (60.4x) 5 3 210 (26.7x) 347 (44.1x) 354
(44.9x) 8 1 20x 30x 27x 7 4 195 (22x) 290 (32.8x) 291 (32.9x) 9 5
247 (16.3x) 395 (26x) 421 (27.8x) 15 6 202 (10.1x) 231 (11.6x) 266
(13.3x) 20 7 415 (9.9x) 629 (14.9x) 606 (14.4x) 42 15 1410 (102.2x)
1390 (100.8x) 1852 (134.3x) 14 8 82 (2.1x) 82 (2.1x) 219 (5.7x) 38
16 332 (15.7x) 384 (18.1x) 600 (28.3x) 21 17 182 (33.5x) 161
(29.6x) 198 (36.5x) 5 18 136 (10.8x) 170 13.5x) 177 (14.1x) 13 10
329 (32.6x) 631 (62.5x) 430 (42.6x) 10 12 212 (14.1x) 453 (30.2x)
352 (23.5x) 15 20 10598 (12.3x 17328 (20.1x) 16328 (18.9x) 864 21
241 (46x) 526 (100.3x) 484 (92.2x) 5 22 222 (83.1x) 446 (166.7x)
313 (117x) 3 19 251 (69.1x) 478 (131.7x) 319 (88x) 4 23 155 (60.4x)
343 (134x) 257 (100.2x) 3 24 133 (26x) 261 (50.9x) 193 (37.6x) 5 25
516 (50.9x) 938 (92.6x) 830 (81.9x) 10 26 159 (31.4x) 345 (68.3x)
270 (53.4x) 5 27 417 (63.5x) 700 (106.6x) 745 (113.5x) 7 28 111
(30.7x) 187 (51.5x) 181 (50.1x) 4 29 464 (60.9x) 808 (106x) 520
(68.2x) 8 30 88 (27.5x) 181 (56.5x) 141 (44.1x) 3 31 206 (19.8x)
362 (34.7x) 221 (21.1x) 10 32 47 (22.6x) 83 (39.8x) 85 (41x) 2 33
110 (56.4x) 226 (116.2x) 169 (86.7x) 2 34 20 (15.5x) 28 (21.5x) 34
(25.7x) 1 35 77 (42.3x) 140 (76.2x) 128 (7.0x) 2 IC50 (nM) Compound
ID HDAC1 HDAC2 HDAC3 HDAC6 36 40 (17.5x) 71 (31.2x) 75 (33x) 2 37
89 (17.8x) 127 (25.4x) 149 (29.8x) 5 38 470 (67.5x) 720 (103.5x)
891 (128.1x) 7 39 263 (62.6x) 387 (92.2x) 495 (117.9x) 4 40 38
(19.8x) 56 (28.7x) 70 (36.2x) 2 41 214 (48.9x) 259 (59.2x) 326
(74.5x) 4 42 78 (19.5x) 97 (24.1x) 130 (32.3x) 4 43 90 (22.8x) 120
(30.3x) 107 (27.1x) 4 44 173 (15.9x) 253 (23.2x) 307 (28.2x) 11 45
377 (106.9x) 523 (148.5x) 506 (143.4x) 4 46 40 (21.4x) 53 (28.4x)
58 (31.2x) 2 47 223 (55.8x) 346 (86.7x) 300 (75.1x) 4 48 363
(48.8x) 601 (80.8x) 401 (53.9x) 7 49 219 (99.5x) 338 (154x) 303
(138.1x) 2 50 1166 (288.1x) 1516 (374.6x) 2214 (547.1x) 4 51 468
(41.2x) 624 (55x) 382 (33.6x) 11 52 185 (60.9x) 248 (81.6x) 254
(83.6x) 3 53 24 (14.9x) 30 (19.2x) 43 (27.2x) 2 54 1370 (64.4x)
2105 (98.9x) 1568 (73.7x) 21 55 460 (60.2x) 587 (76.8x) 631 (82.6x)
8 57 526 (86x) 713 (116.6x) 901 (147.4x) 6 58 552 (60.8x) 701
(77.2x) 1085 (119.5x) 9 59 607 (15.3x) 815 (20.5x) 873 (22x) 40 60
129 (53.5x) 142 (58.9x) 237 (98.1x) 2 61 1336 (35.9x) 1540 (41.4x)
3263 (87.7x) 37 62 35 (22.5x) 56 (36x) 55 (35.2.x) 2 63 1445
(77.6x) 1798 (96.5x) 1746 (93.7x) 19 65 1128 (61x) 1701 (91.9x)
7482 (404.4x) 19 66 39 (29.1x) 39 (29.2x) 47 (35.2x) 1 67 61
(26.1x) 79 (33.8x) 100 (42.8x) 2 68 761 (254.8x) 1083 (362.4x) 1480
(495.3x) 3 69 204 (41.5x) 240 (48.7x) 299 (60.9x) 5 70 645 (217.2x)
972 (327.2x) 1313 (441.9x) 3 71 500 (202.9x) 912 (370x) 1260
(511.4x) 2 72 5752 (306.9x) 8006 (427.2x) 12297 (656.2x) 19 73 94
(60x) 128 (81.9x) 219 (139.5x) 2 74 889 (147.5x) 1015 (168.4x) 2376
(394.2x) 6 75 1142 (374.2x) 966 (316.3x) 1882 (616.6x) 3 76 624
(153.3x) 609 (149.7x) 1058 (260x) 4 77 328 (74x) 340 (76.9x) 434
(98.1x) 4 78 598 (137.3x) 669 (153.6x) 1327 (304.6x) 4 79 1268
(221.6x) 1830 (319.8x) 4676 (817.2x) 6 80 85 (40.4x) 127 (60.2x)
212 (100.3x) 2 81 47 (18.3x) 78 (30.2x) 120 (46.4x) 3 82 16 (9.9x)
18 (11.2x) 22 (13.6x) 2 83 447 (139.2x) 452 (140.9x) 928 (289x) 3
84 2226 (302.2x) 2971 (403.4x) 4515 (613x) 7 85 67 (18.1x) 96
(26.2x) 135 (36.9x) 4 86 788 (60.3x) 1292 (98.9x) 2891 (221.2x) 13
87 85 (45.4x) 116 (61.6x) 233 (124.1x) 2 88 711 (215.2x) 978
(296.1x) 2243 (679.1x) 3 89 223 (64.4x) 270 (78.1x) 549 (158.8x) 3
90 27 (19.9x) 36 (26.2x) 41 (30.2x) 1 91 160 (72.3x) 288 (130.2x)
374 (169.4x) 2 92 1359 (41.3x) 2235 (68x) 4422 (134.4x) 33 93 80
(25.9x) 107 (34.7x) 157 (51x) 3 94 22 (12x) 32 (17.4x) 50 (27.4x) 2
95 586 (176.7x) 722 (217.7x) 2272 (685x) 3 96 2341 (271.8x) 2465
(286.2x) 5709 (662.8x) 9 97 19 (8.3x) 28 (12.1x) 37 (16.4x) 2 98
147 (39.9x) 239 (64.8x) 4252 (115.2x) 4 99 200 (47.3x) 320 (75.6x)
345 (81.5x) 4 100 3206 (339.9x) 4761 (504.7x) 13474 (1428.4x) 9 101
2123 (283.5x) 2570 (343.2x) 11223 (1498.8x) 7 102 13 (7.4x) 21
(11.3x) 33 (18x) 2 103 578 (12.6x) 832 (18.1x) 1098 (23.9x) 46 104
813 (16.7x) 1174 (24.1x) 1172 (24x) 49 105 214 (54.5x) 361 (92.1x)
361 (92x) 4 106 92 (25.3x) 133 (36.5x) 174 (47.7x) 4 107 60 (2.7x)
9 (3.7x) 19 (8x) 2 108 219 (42.1x) 313 (60.3x) 355 (68.3x) 5 109
324 (79.5x) 385 (94.3x) 843 (206.7x) 4 110 1409 (34.5x) 1631
(39.9x) 3460 (84.7x) 41 111 911 (48x) 767 (40.5x) 1327 (70x) 19 120
38 (9.8x) 53 (13.5x) 128 (32.8x) 4 121 27 (12.4x) 32 (14.5x) 99
(44.8x) 2 122 893 (44.8x) 898 (45x) 3944 (197.8x) 20 123 13 (4.8x)
12 (4.3x) 26 (9.6x) 3 124 5 (3.6x) 6 (3.9x) 8 (5.9x) 1 125 18
(8.7x) 19 (9x) 57 (27.1x) 2 126 7 (3.2x) 9 (3.9x) 22 (9.8x) 2 127
50 (23.9x) 57 (27.1x) 119 (56.1x) 2 128 16 (8.1x) 17 (8.9x) 49
(24.8x) 2 129 43 (12.7x) 52 (15.4x) 124 (36.6x) 3 130 140 (43.3x)
159 (48.9x) 521 (160.9x) 3 131 17 (4.8x) 19 (5.5x) 62 (18x) 3 132
95 (24.4x) 96 (24.4x) 316 (80.8x) 4 133 594 (188x) 483 (152.9x) 727
(229.8x) 3 134 47 (14x) 40 (11.8x) 99 (29.2x) 3 135 149 (22.4x) 118
(17.7x) 344 (51.8x) 7 136 45 (13.3x) 40 (11.9x) 107 (31.4x) 3 137
27 (7.1x) 21 (5.5x) 70 (18.5x) 4 138 50 (6.2x) 49 (6x) 113 (13.9x)
8 139 386 (90.9x) 479 (112.8x) 782 (184x) 4 140 55 (8x) 62 (9x) 128
(18.5x) 7 141 13 (3.2x) 17 (4.2x) 54 (13x) 4 142 17 (3x) 23 (4x) 64
(11x) 6 143 12 (3.4x) 17 (5x) 29 (8.4x) 4 144 25 (7.9x) 34 (10.8x)
58 (18.3x) 3 145 104 (18.1x) 135 (23.5x) 384 (66.6x) 6 146 16
(5.2x) 22 (7x) 57 (18.6x) 3 147 64 (18.2x) 73 (20.9x) 189 (54x) 3
148 583 (182.9x) 615 (193.1x) 2157 (677.2x) 3 149 339 (80.1x) 330
(78x) 1324 (312.9x) 4 150 320 (66.1x) 330 (68.4x) 1777 (367.7x) 5
151 584 (176.6x) 476 (143.8x) 1373 (414.9x) 3 152 248 (11.4x) 220
(10.1x) 649 (29.9x) 4 153 528 (64.4x) 491 (59.9x) 1255 (153.1x) 8
154 1372 (263.9x) 1600 (307.8x) 3365 (647.4x) 5 155 262 (37.9x) 231
(34x) 1219 (176x) 7
INCORPORATION BY REFERENCE
[0338] The contents of all references (including literature
references, issued patents, published patent applications, and
co-pending patent applications) cited throughout this application
are hereby expressly incorporated herein in their entireties by
reference. Unless otherwise defined, all technical and scientific
terms used herein are accorded the meaning commonly known to one
with ordinary skill in the art.
EQUIVALENTS
[0339] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, many
equivalents of the specific embodiments of the invention described
herein. Such equivalents are intended with be encompassed by the
following claims.
* * * * *