U.S. patent application number 11/735311 was filed with the patent office on 2007-08-09 for heterocyclic compounds.
This patent application is currently assigned to Janssen Pharmaceutica, N.V.. Invention is credited to Hui Cai, Nicholas I. Carruthers, Curt A. Dvorak, James P. Edwards, Annette K. Kwok, Jianmei Wei.
Application Number | 20070185131 11/735311 |
Document ID | / |
Family ID | 31978666 |
Filed Date | 2007-08-09 |
United States Patent
Application |
20070185131 |
Kind Code |
A1 |
Cai; Hui ; et al. |
August 9, 2007 |
Heterocyclic Compounds
Abstract
Certain thienopyrrolyl and furanopyrrolyl compounds are
disclosed as useful to treat or prevent disorders and conditions
mediated by the histamine H.sub.4 receptor, including allergic
rhinitis.
Inventors: |
Cai; Hui; (San Diego,
CA) ; Carruthers; Nicholas I.; (Poway, CA) ;
Dvorak; Curt A.; (San Diego, CA) ; Edwards; James
P.; (San Diego, CA) ; Kwok; Annette K.;
(Concord, NH) ; Wei; Jianmei; (San Diego,
CA) |
Correspondence
Address: |
PHILIP S. JOHNSON;JOHNSON & JOHNSON
ONE JOHNSON & JOHNSON PLAZA
NEW BRUNSWICK
NJ
08933-7003
US
|
Assignee: |
Janssen Pharmaceutica, N.V.
|
Family ID: |
31978666 |
Appl. No.: |
11/735311 |
Filed: |
April 13, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10656059 |
Sep 5, 2003 |
7226938 |
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11735311 |
Apr 13, 2007 |
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60408723 |
Sep 6, 2002 |
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Current U.S.
Class: |
514/254.08 ;
544/373 |
Current CPC
Class: |
A61P 17/00 20180101;
A61P 37/00 20180101; A61P 9/10 20180101; A61P 11/06 20180101; A61P
43/00 20180101; A61P 25/00 20180101; C07D 495/04 20130101; A61P
29/00 20180101; C07D 491/04 20130101; A61P 17/06 20180101; A61P
1/04 20180101; A61P 37/08 20180101 |
Class at
Publication: |
514/254.08 ;
544/373 |
International
Class: |
A61K 31/496 20060101
A61K031/496; C07D 491/02 20060101 C07D491/02 |
Claims
1. A compound of formula (I): ##STR26## Y is O Z is O or S; n is 1
or 2; m is 1 or 2; n+m is 2 or 3; R.sup.1 is H or C.sub.1-6alkyl;
R.sup.2 is H, F, Cl, Br or C.sub.1-6alkyl; R.sup.3 and R.sup.4 are,
independently, H, C.sub.1-4alkyl, C.sub.3-6cycloalkyl,
C.sub.1-4alkyl(C.sub.3-6cycloalkyl), cyano, --CF.sub.3,
--(CO)NR.sup.pR.sup.q, --(CO)OR.sup.r, --CH.sub.2NR.sup.pR.sup.q or
--CH.sub.2OR.sup.r; where R.sup.p, R.sup.q and R.sup.r are
independently selected from H, C.sub.1-4alkyl, C.sub.3-6cycloalkyl,
phenyl, --C.sub.1-2alkyl(C.sub.3-6cycloalkyl), benzyl or phenethyl,
or R.sup.p and R.sup.q taken together with the nitrogen to which
they are attached, form a 4-7 membered heterocyclic ring with 0 or
1 additional heteroatoms selected from O, S, NH or NC.sub.1-6alkyl,
and where any phenyl or alkyl or cycloalkyl moiety of the foregoing
is optionally and independently substituted with between 1 and 3
substituents selected from C.sub.1-3alkyl, halo, hydroxy, amino,
and C.sub.1-3alkoxy; R.sup.5 and R.sup.6 are, independently, H or
C.sub.1-6alkyl; R.sup.7 is --R.sup.a, --R.sup.bR.sup.a,
--R.sup.e--R.sup.a or --R.sup.e--N(R.sup.c)(R.sup.d), where R.sup.a
is H, cyano, --(C.dbd.O)N(R.sup.c)(R.sup.d),
--C(.dbd.NH)(NH.sub.2), C.sub.1-10alkyl, C.sub.2-8alkenyl,
C.sub.3-8cycloalkyl, C.sub.4-7heterocyclic radical or phenyl, where
the C.sub.4-7heterocyclic radical is attached at a carbon atom and
contains one of O, S, NH or NC.sub.1-4alkyl, and optionally an
additional NH or NC.sub.1-6alkyl in rings of 5 or 6 or 7 members,
where R.sup.b is C.sub.1-8alkylene or C.sub.2-8alkenylene, where
R.sup.e is C.sub.2-8alkylene or C.sub.2-8alkenylene, where R.sup.c
and R.sup.d are each independently H, C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.3-6cycloalkyl or phenyl, or R.sup.c and
R.sup.d taken together with the nitrogen to which they are
attached, form a 4-7 membered heterocyclic ring with 0 or 1
additional heteroatoms selected from O, S, NH or NC.sub.1-6alkyl,
and where any phenyl or alkyl or cycloalkyl moiety of the foregoing
is optionally and independently substituted with between 1 and 3
substituents selected from C.sub.1-3alkyl, halo, hydroxy, amino,
and C.sub.1-3alkoxy; alternatively, R.sup.7 may be taken together
with an adjacent R.sup.4 as well as their carbon and nitrogen of
attachment to form a 5, 6 or 7 membered heterocyclic ring, with 0
or 1 additional heteroatoms selected from O, S, NH or
NC.sub.1-6alkyl, and optionally and independently substituted with
between 1 and 3 substituents selected from C.sub.1-3alkyl, halo,
hydroxy, amino, and C.sub.1-3alkoxy; R.sup.8 and R.sup.9 are,
independently, H, F, Cl, Br, I, C.sub.1-4alkyl, C.sub.1-4alkoxy,
--C.sub.3-6cycloalkyl, --OC.sub.3-6cycloalkyl, --OCH.sub.2Ph,
--CF.sub.3, --OCF.sub.3, --SCF.sub.3, --(C.dbd.O)R.sup.k (wherein
R.sup.k is H, C.sub.1-4alkyl, --OH, phenyl, benzyl, phenethyl or
C.sub.1-6alkoxy), --(N--R.sup.t)(C.dbd.O)R.sup.k (where R.sup.t is
H or C.sub.1-4alkyl), --(N--R.sup.t)SO.sub.2C.sub.1-4alkyl,
--(S.dbd.(O).sub.p)--C.sub.1-4alkyl (wherein p is 0, 1 or 2),
nitro, --SO.sub.2NR.sup.lR.sup.m (wherein R.sup.l and R.sup.m are
independently selected from H, C.sub.1-4alkyl, phenyl, benzyl or
phenethyl, or R.sup.l and R.sup.m taken together with the nitrogen
to which they are attached, form a 4-7 membered heterocyclic ring
with 0 or 1 additional heteroatoms selected from O, S, NH or
NC.sub.1-4alkyl), --(C.dbd.O)NR.sup.lR.sup.m, cyano or phenyl,
where any phenyl or alkyl or cycloalkyl moiety of the foregoing is
optionally and independently substituted with between 1 and 3
substituents selected from C.sub.1-3alkyl, halo, hydroxy, amino,
and C.sub.1-3alkoxy; and enantiomers, diastereomers and
pharmaceutically acceptable salts and esters thereof, with the
following provisos, that R.sup.6 adjacent to N must be H where
R.sup.4 adjacent to N is other than H, that R.sup.7 is not
--CH.sub.2CH.sub.2OH; and that where the core molecule is a
4H-furo, then one of R.sup.4 and R.sup.6 adjacent to N must not be
methyl when the other is hydrogen unless R.sup.6 and R.sup.4 are
taken together to form a bridging moiety.
2. A pharmaceutical composition containing at least one compound of
formula (I) ##STR27## Y is O; Z is O or S; n is 1 or 2; m is 1 or
2; n+m is 2 or 3; R.sup.1 is H or C.sub.1-6alkyl; R.sup.2 is H, F,
Cl, Br or C.sub.1-6alkyl; R.sup.3 and R.sup.4 are, independently,
H, C.sub.1-4alkyl, C.sub.3-6cycloalkyl,
C.sub.1-4alkyl(C.sub.3-6cycloalkyl), cyano, --CF.sub.3,
--(CO)NR.sup.pR.sup.q, --(CO)OR.sup.r, --CH.sub.2NR.sup.pR.sup.q Or
--CH.sub.2OR.sup.r; where R.sup.p, R.sup.q and R.sup.r are
independently selected from H, C.sub.1-4alkyl, C.sub.3-6cycloalkyl,
phenyl, --C.sub.1-2alkyl(C.sub.3-6cycloalkyl), benzyl or phenethyl,
or R.sup.p and R.sup.q taken together with the nitrogen to which
they are attached, form a 4-7 membered heterocyclic ring with 0 or
1 additional heteroatoms selected from O, S, NH or NC.sub.1-6alkyl,
and where any phenyl or alkyl or cycloalkyl moiety of the foregoing
is optionally and independently substituted with between 1 and 3
substituents selected from C.sub.1-3alkyl, halo, hydroxy, amino,
and C.sub.1-3alkoxy; R.sup.5 and R.sup.6 are, independently, H or
C.sub.1-6alkyl; R.sup.7 is --R.sup.a, --R.sup.bR.sup.a,
--R.sup.e--O--R.sup.a or --R.sup.e--N(R.sup.c)(R.sup.d), where
R.sup.a is H, cyano, --(C.dbd.O)N(R.sup.c)(R.sup.d),
--C(.dbd.NH)(NH.sub.2), C.sub.1-10alkyl, C.sub.2-8alkenyl,
C.sub.3-8cycloalkyl, C.sub.4-7heterocyclic radical or phenyl, where
the C.sub.4-7heterocyclic radical is attached at a carbon atom and
contains one of O, S, NH or NC.sub.1-4alkyl, and optionally an
additional NH or NC.sub.1-6alkyl in rings of 5 or 6 or 7 members,
where R.sup.b is C.sub.1-8alkylene or C.sub.2-8alkenylene, where
R.sup.e is C.sub.2-8alkylene or C.sub.2-8alkenylene, where R.sup.c
and R.sup.d are each independently H, C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.3-6cycloalkyl or phenyl, or R.sup.c and
R.sup.d taken together with the nitrogen to which they are
attached, form a 4-7 membered heterocyclic ring with 0 or 1
additional heteroatoms selected from O, S, NH or NC.sub.1-6alkyl,
and where any phenyl or alkyl or cycloalkyl moiety of the foregoing
is optionally and independently substituted with between 1 and 3
substituents selected from C.sub.1-3alkyl, halo, hydroxy, amino,
and C.sub.1-3alkoxy; alternatively, R.sup.7 may be taken together
with an adjacent R.sup.4 as well as their carbon and nitrogen of
attachment to form a 5, 6 or 7 membered heterocyclic ring, with 0
or 1 additional heteroatoms selected from O, S, NH or
NC.sub.1-6alkyl, and optionally and independently substituted with
between 1 and 3 substituents selected from C.sub.1-3alkyl, halo,
hydroxy, amino, and C.sub.1-3alkoxy; R.sup.8 and R.sup.9 are,
independently, H, F, Cl, Br, I, C.sub.1-4alkyl, C.sub.1-4alkoxy,
--C.sub.3-6cycloalkyl, --OC.sub.3-6cycloalkyl, --OCH.sub.2Ph,
--CF.sub.3, --OCF.sub.3, --SCF.sub.3, --(C.dbd.O)R.sup.k (wherein
R.sup.k is H, C.sub.1-4alkyl, --OH, phenyl, benzyl, phenethyl or
C.sub.1-6alkoxy), --(N--R.sup.t)(C.dbd.O)R.sup.k (where R.sup.t is
H or C.sub.1-4alkyl), --(N--R.sup.t)SO.sub.2C.sub.1-4alkyl,
--(S.dbd.(O).sub.p)--C.sub.1-4alkyl (wherein p is 0, 1 or 2),
nitro, --SO.sub.2NR.sup.lR.sup.m (wherein R.sup.l and R.sup.m are
independently selected from H, C.sub.1-4alkyl, phenyl, benzyl or
phenethyl, or R.sup.l and R.sup.m taken together with the nitrogen
to which they are attached, form a 4-7 membered heterocyclic ring
with 0 or 1 additional heteroatoms selected from O, S, NH or
NC.sub.1-4alkyl), --(C.dbd.O)NR.sup.lR.sup.m, cyano or phenyl,
where any phenyl or alkyl or cycloalkyl moiety of the foregoing is
optionally and independently substituted with between 1 and 3
substituents selected from C.sub.1-3alkyl, halo, hydroxy, amino,
and C.sub.1-3alkoxy; and enantiomers, diastereomers and
pharmaceutically acceptable salts and esters thereof, with the
following provisos, that R.sup.6 adjacent to N must be H where
R.sup.4 adjacent to N is other than H, that R.sup.7 is not
--CH.sub.2CH.sub.2OH; and that where the core molecule is a
4H-furo, then one of R.sup.4 and R.sup.6 adjacent to N must not be
methyl when the other is hydrogen unless R.sup.6 and R.sup.4 are
taken together to form a bridging moiety.
3. A method for modulating the H.sub.4 receptor, comprising
exposing said H.sub.4 receptor to an effective amount of at least
one compound of claim 1.
4. A compound selected from the group consisting of
(4H-Furo[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-methanone;
(2-Methyl-4H-furo[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-methanone;
(2,3-Dimethyl-4H-furo[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-metha-
none;
(2-Methyl-4H-furo[3,2-b]pyrrol-5-yl)-piperazin-1-yl-methanone; and
enantiomers, diastereomers and pharmaceutically acceptable salts
and esters thereof.
5. A method for modulating the H.sub.4 receptor, comprising
exposing said H.sub.4 receptor to an effective amount of at least
one compound of claim 4.
6. A pharmaceutical composition containing at least one compound of
claim 4.
Description
[0001] This application is a division of U.S. patent application
Ser. No. 10/656,059, filed Sep. 5, 2003, which in turn claims the
benefit of U.S. provisional patent application Ser. No. 60/408,723,
filed Sep. 6, 2002.
FIELD OF THE INVENTION
[0002] The invention relates to novel, pharmaceutically active,
fused heterocyclic compounds and methods of using them to treat or
prevent disorders and conditions mediated by the histamine H.sub.4
receptor.
BACKGROUND OF THE INVENTION
[0003] Histamine was first identified as a hormone (G. Barger and
H. H. Dale, J. Physiol. (London) 1910, 41:19-59) and has since been
demonstrated to play a major role in a variety of physiological
processes, including the inflammatory "triple response" via H.sub.1
receptors (A. S. F. Ash and H. O. Schild, Br. J. Pharmac.
Chemother. 1966, 27:427-439), gastric acid secretion via H.sub.2
receptors (J. W. Black et al., Nature 1972, 236:385-390), and
neurotransmitter release in the central nervous system via H.sub.3
receptors (J.-M. Arrang et al., Nature 1983, 302:832-837) (for
review see S. J. Hill et al., Pharmacol. Rev. 1997, 49(3):253-278).
All three histamine receptor subtypes have been demonstrated to be
members of the superfamily of G protein-coupled receptors (I. Gantz
et al., Proc. Natl. Acad. Sci. U.S.A. 1991, 88:429-433; T. W.
Lovenberg et al., Mol. Pharmacol. 1999, 55(6):1101-1107; M.
Yamashita et al., Proc. Natl. Acad. Sci. U.S.A. 1991,
88:11515-11519). There are, however, additional functions of
histamine that have been reported, for which no receptor has been
identified. For example, in 1994, Raible et al. demonstrated that
histamine and R-.alpha.-methylhistamine could activate calcium
mobilization in human eosinophils (D. G. Raible et al., Am. J.
Respir. Crit. Care Med. 1994,149:1506-1511). These responses were
blocked by the H.sub.3-receptor antagonist thioperamide. However,
R-.alpha.-methylhistamine was significantly less potent than
histamine, which was not consistent with the involvement of known
H.sub.3 receptor subtypes. Therefore, Raible et al. hypothesized
the existence of a novel histamine receptor on eosinophils that was
non-H.sub.1, non-H.sub.2, and non-H.sub.3. Most recently several
groups (T. Oda et al., J. Biol. Chem. 2000, 275(47):36781-36786; C.
Liu et al., Mol. Pharmacol. 2001, 59(3):420-426; T. Nguyen et al.,
Mol. Pharmacol. 2001, 59(3):427-433; Y. Zhu et al., Mol. Pharmacol.
2001, 59(3):434-441; K. L. Morse et al., J. Pharmacol. Exp. Ther.
2001, 296(3):1058-1066) have identified and characterized a fourth
histamine receptor subtype, the H.sub.4 receptor. This receptor is
a 390 amino acid, seven-transmembrane, G protein-coupled receptor
with approximately 40% homology to the histamine H.sub.3 receptor.
In contrast to the H.sub.3 receptor, which is primarily located in
the brain, the H.sub.4 receptor is expressed at greater levels in
neutrophils and mast cells, among other cells, as reported by Morse
et al. (see above).
[0004] Events that elicit the inflammatory response include
physical stimulation (including trauma), chemical stimulation,
infection, and invasion by a foreign body. The inflammatory
response is characterized by pain, increased temperature, redness,
swelling, reduced function, or a combination of these. Many
conditions, such as allergies, asthma, chronic obstructed pulmonary
disease (COPD), atherosclerosis, and autoimmune diseases, including
rheumatoid arthritis and lupus, are characterized by excessive or
prolonged inflammation. Inhibition of leukocyte recruitment can
provide significant therapeutic value. Inflammatory diseases or
inflammation-mediated diseases or conditions include, but are not
limited to, acute inflammation, allergic inflammation, and chronic
inflammation.
[0005] Mast cell de-granulation (exocytosis) leads to an
inflammatory response that may be initially characterized by a
histamine-modulated wheal and flare reaction. A wide variety of
immunological (e.g., allergens or antibodies) and non-immunological
(e.g., chemical) stimuli may cause the activation, recruitment, and
de-granulation of mast cells. Mast cell activation initiates
allergic (H.sub.1) inflammatory responses, which in turn cause the
recruitment of other effector cells that further contribute to the
inflammatory response. The histamine H2 receptors modulate gastric
acid secretion, and the histamine H3 receptors affect
neurotransmitter release in the central nervous system.
[0006] Examples of textbooks on the subject of inflammation include
J. I. Gallin and R. Snyderman, Inflammation: Basic Principles and
Clinical Correlates, 3.sup.rd Edition, (Lippincott Williams &
Wilkins, Philadelphia, 1999); V. Stvrtinova, J. Jakubovsky and 1.
Hulin, "Inflammation and Fever", Pathophysiology Principles of
Diseases (Textbook for Medical Students, Academic Press, 1995);
Cecil et al., Textbook Of Medicine, 18.sup.th Edition (W.B.
Saunders Company, 1988); and Steadmans Medical Dictionary.
SUMMARY OF THE INVENTION
[0007] The invention features a compound of formula (I): ##STR1##
[0008] Y is O or S; [0009] Z is O or S; [0010] n is 1 or 2; [0011]
m is 1 or 2; [0012] n+m is 2 or 3; [0013] R.sup.1 is H or
C.sub.1-6alkyl; [0014] R.sup.2 is H, F, Cl, Br or C.sub.1-6alkyl;
[0015] R.sup.3 and R.sup.4 are, independently, H, C.sub.1-4alkyl,
C.sub.3-6cycloalkyl, C.sub.1-4alkyl(C.sub.3-6cycloalkyl), cyano,
--CF.sub.3, --(CO)NR.sup.pR.sup.q, --(CO)OR.sup.r,
--CH.sub.2NR.sup.pR.sup.q Or --CH.sub.2OR.sup.r; where R.sup.p,
R.sup.q and R.sup.r are independently selected from H,
C.sub.1-4alkyl, C.sub.3-6cycloalkyl, phenyl,
--C.sub.1-2alkyl(C.sub.3-6cycloalkyl), benzyl or phenethyl, or
R.sup.p and R.sup.q taken together with the nitrogen to which they
are attached, form a 4-7 membered heterocyclic ring with 0 or 1
additional heteroatoms selected from O, S, NH or NC.sub.1-6alkyl,
and where any phenyl or alkyl or cycloalkyl moiety of the foregoing
is optionally and independently substituted with between 1 and 3
substituents selected from C.sub.1-3alkyl, halo, hydroxy, amino,
and C.sub.1-3alkoxy; [0016] R.sup.5 and R.sup.6 are, independently,
H or C.sub.1-6alkyl; [0017] R.sup.7 is --R.sup.a, --R.sup.bR.sup.a,
--R.sup.e--R.sup.a or --R.sup.e--N(R.sup.c)(R.sup.d), where R.sup.a
is H, cyano, --(C.dbd.O)N(R.sup.c)(R.sup.d),
--C(.dbd.NH)(NH.sub.2), C.sub.1-10alkyl, C.sub.2-8alkenyl,
C.sub.3-8cycloalkyl, C.sub.4-7heterocyclic radical or phenyl, where
the C.sub.4-7heterocyclic radical is attached at a carbon atom and
contains one of O, S, NH or NC.sub.1-4alkyl, and optionally an
additional NH or NC.sub.1-6alkyl in rings of 5 or 6 or 7 members,
where R.sup.b is C.sub.1-8alkylene or C.sub.2-8alkenylene, where
R.sup.e is C.sub.2-8alkylene or C.sub.2-8alkenylene, where R.sup.c
and R.sup.d are each independently H, C.sub.1-4alkyl,
C.sub.2-4alkenyl, C.sub.3-6cycloalkyl or phenyl, or R.sup.c and
R.sup.d taken together with the nitrogen to which they are
attached, form a 4-7 membered heterocyclic ring with 0 or 1
additional heteroatoms selected from O, S, NH or NC.sub.1-6alkyl,
and where any phenyl or alkyl or cycloalkyl moiety of the foregoing
is optionally and independently substituted with between 1 and 3
substituents selected from C.sub.1-3alkyl, halo, hydroxy, amino,
and C.sub.1-3alkoxy; [0018] alternatively, R.sup.7 may be taken
together with an adjacent R.sup.4 as well as their carbon and
nitrogen of attachment to form a 5, 6 or 7 membered heterocyclic
ring, with 0 or 1 additional heteroatoms selected from O, S, NH or
NC.sub.1-6alkyl, and optionally and independently substituted with
between 1 and 3 substituents selected from C.sub.1-3alkyl, halo,
hydroxy, amino, and C.sub.1-3alkoxy; [0019] R.sup.8 and R.sup.9
are, independently, H, F, Cl, Br, I, C.sub.1-4alkyl,
C.sub.1-4alkoxy, --C.sub.3-6cycloalkyl, --OC.sub.3-6cycloalkyl,
--OCH.sub.2Ph, --CF.sub.3, --OCF.sub.3, --SCF.sub.3,
--(C.dbd.O)R.sup.k (wherein R.sup.k is H, C.sub.1-4alkyl, --OH,
phenyl, benzyl, phenethyl or C.sub.1-6alkoxy),
--(N--R.sup.t)(C.dbd.O)R.sup.k (where R.sup.t is H or
C.sub.1-4alkyl), --(N--R.sup.t)SO.sub.2C.sub.1-4alkyl,
--(S.dbd.(O).sub.p)--C.sub.1-4alkyl (wherein p is 0, 1 or 2),
nitro, --SO.sub.2NR.sup.lR.sup.m (wherein R.sup.l and R.sup.m are
independently selected from H, C.sub.1-4alkyl, phenyl, benzyl or
phenethyl, or R.sup.l and R.sup.m taken together with the nitrogen
to which they are attached, form a 4-7 membered heterocyclic ring
with 0 or 1 additional heteroatoms selected from O, S, NH or
NC.sub.1-4alkyl), --(C.dbd.O)NR.sup.lR.sup.m, cyano or phenyl,
where any phenyl or alkyl or cycloalkyl moiety of the foregoing is
optionally and independently substituted with between 1 and 3
substituents selected from C.sub.1-3alkyl, halo, hydroxy, amino,
and C.sub.1-3alkoxy; and enantiomers, diastereomers and
pharmaceutically acceptable salts and esters thereof, with the
following provisos, that R.sup.6 adjacent to N must be H where
R.sup.4 adjacent to N is other than H, that R.sup.7 is not
--CH.sub.2CH.sub.2OH; and that where the core molecule is a
4H-furo, then one of R.sup.4 and R.sup.6 adjacent to N must not be
methyl when the other is hydrogen unless R.sup.6 and R.sup.4 are
taken together to form a bridging moiety.
[0020] The invention also features pharmaceutical compositions
containing such compounds and methods of using such compositions in
the treatment or prevention of H.sub.4-mediated diseases and
conditions, particularly those wherein it is desirable to
antagonize the H.sub.4 receptor.
DETAILED DESCRIPTION
[0021] Preferably, Y is S.
[0022] Preferably, Z is O.
[0023] Preferably, n is 1 and m is 1.
[0024] Preferrably, R.sup.1 is selected from the group consisting
of H or methyl.
[0025] Preferrably, R.sup.2 is H.
[0026] Preferrably, R.sup.3 and R.sup.4 are, independently,
selected from the group consisting of
[0027] a) H,
[0028] b) --CH.sub.3, --CH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2CH.sub.3, --CH(CH.sub.3).sub.2, n-butyl, i-butyl,
t-butyl,
[0029] c) cyclopropyl, cyclopentyl, cyclohexyl,
--CH.sub.2cyclopropyl, --CH.sub.2cyclopentyl, --CH.sub.2cyclohexyl,
--CH.sub.2Ocyclopropyl, --CH.sub.2Ocyclopentyl,
--CH.sub.2Ocyclohexyl,
[0030] d) cyano,
[0031] e) trifluoromethyl,
[0032] f) --(C.dbd.O)NH.sub.2, --(C.dbd.O)NHC.sub.1-4alkyl,
--(C.dbd.O)N(C.sub.1-4alkyl).sub.2, --(C.dbd.O)NHphenyl,
--(C.dbd.O)pyrrolidin-1-yl, --(C.dbd.O)imidazolidin-1-yl,
--(C.dbd.O)pyrazolidin-1-yl, (C.dbd.O)piperidin-1-yl,
--(C.dbd.O)piperazin-1-yl, --(C.dbd.O)morpholin-4-yl,
--(C.dbd.O)thiomorpholin-4-yl,
[0033] g) --COOH, --COOCH.sub.3, --COOCH.sub.2CH.sub.3,
--COOphenyl, --COObenzyl,
[0034] h) --CH.sub.2NH.sub.2, --CH.sub.2NHC.sub.1-4alkyl,
--CH.sub.2N(C.sub.1-4alkyl).sub.2, --CH.sub.2NHphenyl,
--CH.sub.2NHbenzyl, --CH.sub.2pyrrolidin-1-yl,
--CH.sub.2imidazolidin-1-yl, --CH.sub.2pyrazolidin-1-yl,
--CH.sub.2piperidin-1-yl, --CH.sub.2piperazin-1-yl,
--CH.sub.2morpholin-4-yl, --CH.sub.2thiomorpholin-4-yl,
[0035] i) --CH.sub.2OH, --CH.sub.2CH.sub.2OH,
--CH.sub.2CH.sub.2CH.sub.2OH, --CH.sub.2OCH.sub.3,
--CH.sub.2OCH.sub.2CH.sub.3, --CH.sub.2OCH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2OCH(CH.sub.3).sub.2, --CH.sub.2O-n-butyl,
--CH.sub.2O-1-butyl, --CH.sub.2O-t-butyl, --CH.sub.2O phenyl,
--CH.sub.2Obenzyl and --CH.sub.2OCH.sub.2cyclopropyl.
[0036] Most preferrably, R.sup.3 and R.sup.4 are, independently, H
or --CH.sub.3.
[0037] Preferrably, R.sup.5 and R.sup.6 are, independently,
selected from the group consisting of H and methyl.
[0038] Most preferrably, R.sup.5 and R.sup.6 are H.
[0039] Preferrably, R.sup.7 is selected from the group consisting
of
[0040] a) H, --CH.sub.2CH.sub.2CH.sub.2OH,
[0041] b) cyano,
[0042] c) --(C.dbd.O)NH.sub.2, --(C.dbd.O)NHC.sub.1-4alkyl,
--(C.dbd.O)N(C.sub.1-4alkyl).sub.2, --(C.dbd.O)NHphenyl,
--(C.dbd.O)pyrrolidin-1-yl, --(C.dbd.O)imidazolidin-1-yl,
--(C.dbd.O)pyrazolidin-1-yl, --(C.dbd.O)piperidin-1-yl,
--(C.dbd.O)piperazin-1-yl, --(C.dbd.O)morpholin-4-yl,
--(C.dbd.O)thiomorpholin-4-yl, --CH.sub.2(C.dbd.O)NH.sub.2,
--CH.sub.2(C.dbd.O)NHC.sub.1-4alkyl,
--CH.sub.2(C.dbd.O)N(C.sub.1-4alkyl).sub.2,
--CH.sub.2(C.dbd.O)NHphenyl, --CH.sub.2(C.dbd.O)pyrrolidin-1-yl,
--CH.sub.2(C.dbd.O)imidazolidin-1-yl,
--CH.sub.2(C.dbd.O)pyrazolidin-1-yl,
--CH.sub.2(C.dbd.O)piperidin-1-yl,
--CH.sub.2(C.dbd.O)piperazin-1-yl,
--CH.sub.2(C.dbd.O)morpholin-4-yl,
--CH.sub.2(C.dbd.O)thiomorpholin-4-yl,
--CH.sub.2CH.sub.2O(C.dbd.O)NH.sub.2,
--CH.sub.2CH.sub.2O(C.dbd.O)NHC.sub.1-4alkyl,
--CH.sub.2CH.sub.2O(C.dbd.O)N(C.sub.1-4alkyl).sub.2,
--CH.sub.2CH.sub.2O(C.dbd.O)NHphenyl,
--CH.sub.2CH.sub.2O(C.dbd.O)pyrrolidin-1-yl,
--CH.sub.2CH.sub.2O(C.dbd.O)imidazolidin-1-yl,
--CH.sub.2CH.sub.2O(C.dbd.O)pyrazolidin-1-yl,
--CH.sub.2CH.sub.2O(C.dbd.O)piperidin-1-yl,
--CH.sub.2CH.sub.2O(C.dbd.O)piperazin-1-yl,
--CH.sub.2CH.sub.2O(C.dbd.O)morpholin-4-yl,
--CH.sub.2CH.sub.2O(C.dbd.O)thiomorpholin-4-yl,
[0043] d) --C(.dbd.NH)(NH.sub.2),
--CH.sub.2C(.dbd.NH)(NH.sub.2),
[0044] e) --CH.sub.3, --CH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2CH.sub.3, --CH(CH.sub.3).sub.2, n-butyl, i-butyl,
t-butyl, --CH.sub.2CH.sub.2OCH.sub.3,
--CH.sub.2CH.sub.2OCH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2OCH.sub.2CH.sub.2CH.sub.3,
--CH.sub.2CH.sub.2OCH(CH.sub.3).sub.2, --CH.sub.2CH.sub.2O-n-butyl,
--CH.sub.2CH.sub.2O-1-butyl, --CH.sub.2CH.sub.2O-t-butyl,
[0045] f), --CH.sub.2CH.dbd.CH.sub.2,
[0046] g) cyclopropyl, cyclopentyl, cyclohexyl,
--CH.sub.2cyclopropyl, --CH.sub.2cyclopentyl, --CH.sub.2cyclohexyl,
--CH.sub.2CH.sub.2Ocyclopropyl, --CH.sub.2CH.sub.2Ocyclopentyl,
--CH.sub.2CH.sub.2Ocyclohexyl,
[0047] h) pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl,
piperazinyl, morpholinyl, thiomorpholinyl, --CH.sub.2pyrrolidinyl,
--CH.sub.2imidazolidinyl, --CH.sub.2pyrazolidinyl,
--CH.sub.2piperidinyl, --CH.sub.2piperazinyl,
--CH.sub.2morpholinyl, --CH.sub.2thiomorpholinyl,
[0048] i) --CH.sub.2CH.sub.2NH.sub.2,
--CH.sub.2CH.sub.2NHC.sub.1-4alkyl,
--CH.sub.2CH.sub.2N(C.sub.1-4alkyl).sub.2,
--CH.sub.2CH.sub.2NHphenyl, --CH.sub.2CH.sub.2pyrrolidin-1-yl,
--CH.sub.2CH.sub.2imidazolidin-1-yl,
--CH.sub.2CH.sub.2pyrazolidin-1-yl,
--CH.sub.2CH.sub.2piperidin-1-yl, --CH.sub.2CH.sub.2piperazin-1-yl,
--CH.sub.2CH.sub.2morpholin-4-yl,
--CH.sub.2CH.sub.2thiomorpholin-4-yl,
[0049] j) phenyl, benzyl, phenethyl and benzyloxymethyl.
[0050] Most preferrably, R.sup.7 is selected from the group
consisting of H and --CH.sub.3.
[0051] Preferred R.sup.7 taken together with an adjacent R.sup.4 as
well as their carbon and nitrogen of attachment are
pyrrolidin-1,2-yl, imidazolidin-1,2-yl, imidazolidin-1,5-yl,
pyrazolidin-1,5-yl, piperidin-1,2-yl, piperazin-1,2-yl,
morpholin-4,5-yl and thiomorpholin-4,5-yl.
[0052] Most preferred R.sup.7 taken together with an adjacent
R.sup.4 as well as their carbon and nitrogen of attachment are
pyrrolidin-1,2-yl and piperidin-1,2-yl.
[0053] Preferrably, R.sup.8 and R.sup.9 are, independently,
selected from the group consisting of H, --F, --Cl, --Br, --I,
--CH.sub.3, --CH.sub.2CH.sub.3, --OCH.sub.3, --OCH.sub.2CH.sub.3,
--OCH(CH.sub.3).sub.2, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, --Ocyclopentyl, --Ocyclohexyl, --CF.sub.3, --OCF.sub.3,
--SCF.sub.3, --COOH, --COOCH.sub.3, --COOCH.sub.2CH.sub.3,
--C(O)CH.sub.3, --NHCOCH.sub.3, --NCH.sub.3COCH.sub.3,
--NHSO.sub.2CH.sub.3, --NCH.sub.3SO.sub.2CH.sub.3, --SOCH.sub.3,
--SO.sub.2CH.sub.3, --NO.sub.2, --SO.sub.2NH.sub.2,
--SO.sub.2NHCH.sub.3, --SO.sub.2N(CH.sub.3).sub.2, --C(O)NH.sub.2,
--C(O)N(CH.sub.3).sub.2, --C(O)NH(CH.sub.3), --CN and phenyl.
[0054] Most preferrably, R.sup.8 and R.sup.9 are, independently,
selected from the group consisting of hydrogen, methyl, chloro and
bromo. Further, it is most preferred that one or both of R.sup.8
and R.sup.9 are not hydrogen.
[0055] The "pharmaceutically acceptable salts and esters thereof"
refer to those salt and ester forms of the compounds of the present
invention that would be apparent to the pharmaceutical chemist,
i.e., those that are non-toxic and that would favorably affect the
pharmacokinetic properties of said compounds of the present
invention. Those compounds having favorable pharmacokinetic
properties would be apparent to the pharmaceutical chemist, i.e.,
those that are non-toxic and that possess such pharmacokinetic
properties to provide sufficient palatability, absorption,
distribution, metabolism and excretion. Other factors, more
practical in nature, that are also important in the selection are
cost of raw materials, ease of crystallization, yield, stability,
hygroscopicity, and flowability of the resulting bulk drug. In
addition, acceptable salts of carboxylates include sodium,
potassium, calcium and magnesium. Examples of suitable cationic
salts include hydrobromic, hydroiodic, hydrochloric, perchloric,
sulfuric, maleic, fumaric, malic, tartatic, citric, benzoic,
mandelic, methanesulfonic, hydroethanesulfonic, benzenesulfonic,
oxalic, pamoic, 2-naphthalenesulfonic, p-toluenesulfonic,
cyclohexanesulfamic and saccharic. Examples of suitable esters
include such esters where one or more carboxyl substituents is
replaced with p-methoxybenzyloxycarbonyl,
2,4,6-trimethylbenzyloxycarbonyl, 9-anthryloxycarbonyl,
CH.sub.3SCH.sub.2COO--, tetrahydrofur-2-yloxycarbonyl,
tetrahydropyran-2-yloxycarbonyl, fur-2-uloxycarbonyl, benzoyl
methoxycarbonyl, p-nitrobenzyloxycarbonyl, 4-pyridyl
methoxycarbonyl, 2,2,2-trichloroethoxycarbonyl,
2,2,2-tribromoethoxycarbonyl, t-butyloxycarbonyl,
t-amyloxycarbonyl, diphenylmethoxycarbonyl,
triphenylmethoxycarbonyl, adamantyloxycarbonyl,
2-benzyloxyphenyloxycarbonyl, 4-methylthiophenyloxycarbonyl, or
tetrahydropyran-2-yloxycarbonyl.
[0056] The provisos are based on a failure to find activity in at
least one compound meeting the specifications of each proviso.
[0057] Preferred compounds of Formula I are compounds selected from
the group consisting of: ##STR2##
[0058] Additional preferred compounds of Formula I are compounds
selected from the group consisting of: ##STR3##
[0059] Additional preferred compounds of Formula I are compounds
selected from the group consisting of: ##STR4## ##STR5##
[0060] Still further preferred compounds are made according to the
synthetic methods outlined in Schemes 1-4 where Y is S and selected
from the group sting of:
EX Compound
[0061] 26
(2,3-Dimethyl-6H-thieno[2,3-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-meth-
anone; [0062] 27
(2-Chloro-3-methyl-6H-thieno[2,3-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-
-methanone; [0063] 28
(3-Chloro-2-methyl-6H-thieno[2,3-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-
-methanone; [0064] 29
(2-Bromo-6H-thieno[2,3-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-methanone-
; [0065] 30
(3-Bromo-6H-thieno[2,3-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-methanone-
; [0066] 31
(4-Methyl-piperazin-1-yl)-(2-phenyl-6H-thieno[2,3-b]pyrrol-5-yl)-methanon-
e; [0067] 32
[2-(4-Chloro-phenyl)-6H-thieno[2,3-b]pyrrol-5-yl]-(4-methyl-piperazin-1-y-
l)-methanone; [0068] 33
(3-Bromo-4H-thieno[3,2-b]pyrrol-5-yl)-(3,4-dimethyl-piperazin-1-yl)-metha-
none; [0069] 34
(3,4-Dimethyl-piperazin-1-yl)-(3-methyl-4H-thieno[3,2-b]pyrrol-5-yl)-meth-
anone; [0070] 35
(2-Bromo-3-methyl-4H-thieno[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)--
methanone; [0071] 36
(3-Bromo-2-chloro-4H-thieno[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)--
methanone; [0072] 37
(2,3-Dichloro-4H-thieno[3,2-b]pyrrol-5-yl)-(3-methyl-piperazin-1-yl)-meth-
anone; [0073] 38
(4-Methyl-piperazin-1-yl)-(2-phenyl-4H-thieno[3,2-b]pyrrol-5-yl)-methanon-
e; and [0074] 39
(4-Methyl-piperazin-1-yl)-[2-(4-trifluoromethyl-phenyl)-4H-thieno[3,2-b]p-
yrrol-5-yl]-methanone.
[0075] The following terms are defined below, and by their usage
throughout the disclosure.
[0076] "Alkyl" includes straight chain and branched hydrocarbons
with at least one hydrogen removed to form a radical group. Alkyl
groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl,
t-butyl, 1-methylpropyl, pentyl, isopentyl, sec-pentyl, hexyl,
heptyl, octyl, and so on. Alkyl does not include cycloalkyl.
[0077] "Alkenyl" includes straight chain and branched hydrocarbon
radicals as above with at least one carbon-carbon double bond
(sp.sup.2). Alkenyls include ethenyl (or vinyl), prop-1-enyl,
prop-2-enyl (or allyl), isopropenyl (or 1-methylvinyl), but-1-enyl,
but-2-enyl, butadienyls, pentenyls, hexa-2,4-dienyl, and so on.
Alkenyl does not include cycloalkenyl.
[0078] "Alkoxy" includes a straight chain or branched alkyl group
with a terminal oxygen linking the alkyl group to the rest of the
molecule. Alkoxy includes methoxy, ethoxy, propoxy, isopropoxy,
butoxy, t-butoxy, pentoxy and so on. "Aminoalkyl", "thioalkyl", and
"sulfonylalkyl" are analogous to alkoxy, replacing the terminal
oxygen atom of alkoxy with, respectively, NH (or NR), S, and
SO.sub.2.
[0079] "Cycloalkyl" includes cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl, cyclooctyl, and so on.
[0080] "Halo" includes fluoro, chloro, bromo, and iodo, and
preferably fluoro or chloro.
[0081] "Patient" or "subject" includes mammals such as humans and
animals (dogs, cats, horses, rats, rabbits, mice, non-human
primates) in need of observation, experiment, treatment or
prevention in connection with the relevant disease or condition.
Preferably, the patient is a human.
[0082] "Composition" includes a product comprising the specified
ingredients in the specified amounts as well as any product that
results directly or indirectly from combinations of the specified
ingredients in the specified amounts.
[0083] The compounds as described above may be made according to
processes within the skill of the art and/or that are described in
the schemes and examples that follow. To obtain the various
compounds herein, starting materials may be employed that carry the
ultimately desired substituents though the reaction scheme with or
without protection as appropriate. Alternatively, it may be
necessary to employ, in the place of the ultimately desired
substituent, a suitable group that may be carried through the
reaction scheme and replaced as appropriate with the desired
substituent. ##STR6##
[0084] Referring to Scheme 1, there are disclosed the following
notes and additions. Various R.sup.1 may be obtained from E1 or C1
by treatment with a base and an appropriate alkylating agent. Where
R.sup.2 is halo, it may be obtained by treatment of E1 and C1 with
an appropriate halogenating agent. Where R.sup.2 is alkyl, it may
be obtained by replacing the aldehyde of A1 with a ketone. P may be
an alkyl, aryl or benzyl. Suitable bases include NaOEt, LDA, NaH,
DBU, etc. The conversion of B1 to C1 is thermolytic with typical
temperatures ranging from 80 to 200.degree. C. Suitable solvents
for the conversion of B1 to C1 are xylene, cumene, diphenylether,
etc. Acidic or basic hydrolysis will provide deprotection. In the
case where P is benzyl, hydrogenolysis is also useful for
deprotection. Typical coupling reagents for the conversion of D1 to
E1 include EDCI, HBTU, etc. Typical chlorination agents for the
conversion of D1 to E1 include oxalyl chloride and thionyl
chloride. X is a halogenating agent such as Cl.sub.2,
N-bromosuccinimide, TAS-F, Br.sub.2, N-chlorosuccinimide, etc.
##STR7##
[0085] Referring to Scheme 2, there are disclosed the following
notes and additions. Various R.sup.1 may be obtained from E2 or C2
by treatment with a base and an appropriate alkylating agent. Where
R.sup.2 is halo, it may be obtained by treatment of E2 and C2 with
an appropriate halogenating agent. Where R.sup.2 is alkyl, it may
be obtained by replacing the aldehyde of A2 with a ketone. P may be
an alkyl, aryl or benzyl. Suitable bases include NaOEt, LDA, NaH,
DBU, etc. The conversion of B2 to C2 is thermolytic with typical
temperatures ranging from 80 to 200.degree. C. Suitable solvents
for the conversion of B2 to C2 are xylene, cumene, diphenylether,
etc. Acidic or basic hydrolysis will provide deprotection. In the
case where P is benzyl, hydrogenolysis is also useful for
deprotection. Typical coupling reagents for the conversion of D2 to
E2 include EDCI, HBTU, etc. Typical chlorination agents for the
conversion of D2 to E2 include oxalyl chloride and thionyl
chloride. X is a halogenating agent such as Cl.sub.2,
N-bromosuccinimide, TAS-F, Br.sub.2, N-chlorosuccinimide, etc.
##STR8##
[0086] Referring to Scheme 3, there are disclosed the following
notes and additions. X is a halogenating agent such as Cl.sub.2,
N-bromosuccinimide, TAS-F, Br.sub.2, N-chlorosuccinimide, etc.
##STR9##
[0087] Referring to Scheme 4, there are disclosed the following
notes and additions. Typical bases include n-BuLi, LDA, t-BuLi,
KHMDS.
[0088] The expression of the H.sub.4 receptor in immune cells,
including some leukocytes and mast cells, establishes it as an
important target for therapeutic intervention in a range of
immunological and inflammatory disorders (such as allergic,
chronic, or acute inflammation). Specifically H.sub.4 receptor
ligands are expected to be useful for the treatment or prevention
of various mammalian disease states.
[0089] Thus, according to the invention, the disclosed compounds,
where antagonists of the H.sub.4 receptor, and compositions are
useful for the amelioration of symptoms associated with, the
treatment of, and the prevention of, the following conditions and
diseases: inflammatory disorders, asthma, psoriasis, rheumatoid
arthritis, ulcerative colitis, Crohn's disease, inflammatory bowel
disease, multiple sclerosis, allergic disorders, allergic rhinitis,
dermatological disorders, autoimmune disease, lymphatic disorders,
atherosclerosis, and immunodeficiency disorders. The disclosed
compounds may also be useful as adjuvants in chemotherapy or in the
treatment of itchy skin.
[0090] Aspects of the invention include (a) a pharmaceutical
composition comprising a compound of formula (I), or one or more
preferred compounds as described herein, and a pharmaceutically
acceptable carrier; (b) a packaged drug comprising (1) a
pharmaceutical composition comprising a compound of formula (I) and
a pharmaceutically acceptable carrier, and (2) instructions for the
administration of said composition for the treatment or prevention
of an H.sub.4-mediated disease or condition.
[0091] The invention also provides a method for treating an
H.sub.4-mediated condition in a patient, said method comprising
administering to the patient a pharmaceutically effective amount of
a composition comprising a compound of formula (I) and other
disclosed or preferred compounds. For example, the invention
features a method for treating an H.sub.4 mediated condition in a
patient, said method comprising administering to the patient a
pharmaceutically effective H.sub.4-antagonizing amount of a
composition comprising a compound of formula (I).
[0092] The effect of an antagonist may also be produced by an
inverse agonist. Inverse agonism describes the property of a
compound to actively turn off a receptor that displays constitutive
activity. Constitutive activity can be identified in cells that
have been forced to over-express the human H.sub.4 receptor.
Constitutive activity can be measured by examining cAMP levels or
by measuring a reporter gene sensitive to cAMP levels after a
treatment with a cAMP-stimulating agent such as forskolin. Cells
that over-express H.sub.4 receptors will display lower cAMP levels
after forskolin treatment than non-expressing cells. Compounds that
behave as H.sub.4 agonists will dose-dependently lower
forskolin-stimulated cAMP levels in H.sub.4-expressing cells.
Compounds that behave as inverse H.sub.4 agonists will
dose-dependently stimulate cAMP levels in H.sub.4-expressing cells.
Compounds that behave as H.sub.4 antagonists will block either
H.sub.4 agonist-induced inhibition of cAMP or inverse H.sub.4
agonist-induced increases in cAMP.
[0093] Further embodiments of the invention include disclosed
compounds that are inhibitors of a mammalian histamine H.sub.4
receptor function, inhibitors of inflammation or inflammatory
responses in vivo or in vitro, modulators of the expression of a
mammalian histamine H.sub.4 receptor protein, inhibitors of
polymorphonuclear leukocyte activation in vivo or in vitro, or
combinations of the above, and corresponding methods of treatment,
prophylaxis, and diagnosis comprising the use of a disclosed
compound.
[0094] Those skilled in the art will be able to determine,
according to known methods, the appropriate dosage for a patient,
taking into account factors such as age, weight, general health,
the type of symptoms requiring treatment, and the presence of other
medications. In general, an effective amount will be between 0.01
and 1000 mg/kg per day, preferably between 0.5 and 300 mg/kg body
weight, and daily dosages will be between 10 and 5000 mg for an
adult subject of normal weight. Capsules, tablets or other
formulations (such as liquids and film-coated tablets) may be of
between 0.5 and 200 mg, such as 1, 3, 5, 10, 15, 25, 35, 50 mg, 60
mg, and 100 mg and can be administered according to the disclosed
methods.
[0095] Dosage unit forms include tablets, capsules, pills, powders,
granules, aqueous and nonaqueous oral solutions and suspensions,
and parenteral solutions packaged in containers adapted for
subdivision into individual doses. Dosage unit forms can also be
adapted for various methods of administration, including controlled
release formulations, such as subcutaneous implants. Administration
methods include oral, rectal, parenteral (intravenous,
intramuscular, subcutaneous), intracisternal, intravaginal,
intraperitoneal, intravesical, local (drops, powders, ointments,
gels or cream), and by inhalation (a buccal or nasal spray).
[0096] Parenteral formulations include pharmaceutically acceptable
aqueous or nonaqueous solutions, dispersion, suspensions,
emulsions, and sterile powders for the preparation thereof.
Examples of carriers include water, ethanol, polyols (propylene
glycol, polyethylene glycol), vegetable oils, and injectable
organic esters such as ethyl oleate. Fluidity can be maintained by
the use of a coating such as lecithin, a surfactant, or maintaining
appropriate particle size. Carriers for solid dosage forms include
(a) fillers or extenders, (b) binders, (c) humectants, (d)
disintegrating agents, (e) solution retarders, (f) absorption
accelerators, (g) adsorbants, (h) lubricants, (i) buffering agents,
and (k) propellants.
[0097] Compositions may also contain adjuvants such as preserving,
wetting, emulsifying, and dispensing agents; antimicrobial agents
such as parabens, chlorobutanol, phenol, and sorbic acid; isotonic
agents such as a sugar or sodium chloride; absorption-prolonging
agents such as aluminum monostearate and gelatin; and
absorption-enhancing agents.
EXAMPLES
[0098] General Synthetic Procedures
Procedure A: Annulation of Aldehyde with Ethyl Azidoacetate
[0099] A solution of aldehyde A1, A2 or A3 (1 equiv) and ethyl
azidoacetate (4 equiv) was added dropwise to a solution of NaOEt (4
equiv) in EtOH (0.15 M) at 0.degree. C. The reaction mixture was
stirred at 0.degree. C. for 1 h and at room temperature for an
additional 1 h. The reaction mixture was then poured into satd aq
NH.sub.4Cl and extracted with ether. The combined organics were
dried (Na.sub.2SO.sub.4) and concentrated in vacuo. The residue was
purified by silica gel column chromatography to provide the desired
acrylate.
[0100] A solution of the resultant acrylate in xylene (0.2 M) was
heated at 145.degree. C. for 10-60 min and then allowed to cool to
room temperature. The xylene solution was either cooled further to
induce product crystallization or directly subjected to silica gel
column chromatography to obtain the desired annulation product.
Procedure B: Ester Hydrolysis
[0101] A solution (0.2 M) of the ethyl ester (1 equiv, from
Procedure A) and LiOH (5 equiv) in THF/MeOH/H.sub.2O (3:1:1) was
heated at 65.degree. C. overnight, cooled to room temperature,
acidified with 2 N HCl, and extracted with EtOAc. The organic layer
was separated, dried over Na.sub.2SO.sub.4, and concentrated to
give the desired crude acid, which was taken to the next step
without further purification.
Procedure C: Amide Formation Using
1-(3-Dimethylaminopropyl)-3-ethylcarbodimide Hydrochloride
(EDCI)
[0102] A mixture of acid (1 equiv, from Procedure B), amine (1.5
equiv) and EDCI (2.0 equiv) in CH.sub.2Cl.sub.2 (0.2 M) was stirred
at room temperature overnight and then partitioned between
CH.sub.2Cl.sub.2 and satd aq NaHCO.sub.3. The organic layer was
separated, washed with H.sub.2O, dried over Na.sub.2SO.sub.4, and
concentrated. The crude product was further purified by silica gel
column chromatography.
Procedure D: Amide Formation via Acyl Chloride Intermediate
[0103] A mixture of acid (1 equiv, from Procedure B) in
CH.sub.2Cl.sub.2 (0.5 M) was treated at 0.degree. C. with oxalyl
chloride (1.2 equiv) followed by 1-2 drops of DMF. The reaction
mixture was stirred at 0.degree. C. for 30 min then slowly warmed
to room temperature and stirred for an additional 1 h. All
volatiles were removed to provide the crude acyl chloride.
[0104] The resultant acyl chloride was treated with amine (5.0
equiv) in CH.sub.2Cl.sub.2 (0.2 M) and allowed to stir at room
temperature for 3 h. The reaction mixture was partitioned between
CH.sub.2Cl.sub.2 and satd aq NaHCO.sub.3. The organic layer was
separated, washed with H.sub.2O, dried over Na.sub.2SO.sub.4, and
concentrated. The crude product was further purified with silica
gel column chromatography.
General Analytical Procedures
[0105] NMR spectra were obtained on either a Bruker model DPX400
(400 MHz) or DPX500 (500 MHz) spectrometer. The format of the 1H
NMR data below is: chemical shift in ppm down field of the
tetramethylsilane reference (multiplicity, coupling constant J in
Hz, integration).
[0106] Mass spectra were obtained on a Hewlett Packard (Agilent)
series 1100 MSD using electrospray ionization (ESI) in either
positive or negative mode as indicated. The "mass calculated" for a
molecular formula is the monoisotopic mass of the compound.
Silica Gel Column Chromatography:
[0107] Normal-phase column chromatography was accomplished using an
ISCO Foxy 200 system employing one of the following commercially
available prepacked columns: ISCO Redisep (SiO.sub.2, 10 g, 12 g,
35 g, 40 g, or 120 g).
Example 1
[0108] ##STR10##
(4-Methyl-piperazin-1-yl)-(6H-thieno[2,3-b]pyrrol-5-yl)-methanone
[0109] A. 6H-Thieno[2,3-b]pyrrole-5-carboxylic acid ethyl ester.
Thiophene-3-carbaldehyde (2.24 g, 20 mmol) was annulated according
to Procedure A to provide the title compound (1.2 g, 31%) as a
white solid. TLC (silica, 20% EtOAc/hexanes): R.sub.f=0.50. .sup.1H
NMR (CDCl.sub.3, 400 MHz): 10.30 (br s, 1H), 7.10 (d, J=1.9 Hz,
1H), 6.96 (d, J=5.4 Hz, 1H), 6.87 (d, J=5.4 Hz, 1H), 4.39 (q, J=7.1
Hz, 2H), 1.38 (t, J=7.1 Hz, 3H).
[0110] B. 6H-Thieno[2,3-b]pyrrole-5-carboxylic acid.
6H-Thieno[2,3-b]pyrrole-5-carboxylic acid ethyl ester (835 mg, 4.3
mmol) was hydrolyzed according to Procedure B to provide the crude
acid as a pale-yellow solid. .sup.1H NMR (CD.sub.3OD, 400 MHz):
7.02 (s, 1H), 6.96 (s, 1H), 6.95 (s, 1H).
[0111] C.
(4-Methyl-piperazin-1-yl)-(6H-thieno[2,3-b]pyrrol-5-yl)-methano-
ne. 6H-Thieno[2,3-b]pyrrole-5-carboxylic acid (60 mg, 0.35 mmol)
was coupled with N-methylpiperazine according to Procedure C to
provide the title compound (44 mg, 50%) as a light yellow solid.
TLC (silica, 10% MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.4. MS
(electrospray): exact mass calculated for
C.sub.12H.sub.15N.sub.3OS, 249.09; m/z found, 250.1 [M+H].sup.+.
.sup.1H NMR (CD.sub.3OD, 400 MHz, TFA salt): 6.97 (s, 1H), 6.96 (s,
1H), 6.85 (s, 1H), 4.20-3.10 (m, 8H), 2.96 (s, 3H).
Example 2
[0112] ##STR11##
(Hexahydro-pyrrolo[1,2-a]pyrazin-2-yl)-(6H-thieno[2,3-b]pyrrol-5-yl)-metha-
none
[0113] 6H-Thieno[2,3-b]pyrrole-5-carboxylic acid (60 mg, 0.35 mmol)
was coupled with octahydro-pyrrolo[1,2-a]pyrazine according to
Procedure C to provide the title compound (34 mg, 35%) as a light
yellow solid. TLC (silica, 10% MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.4.
MS (electrospray): exact mass calculated for
C.sub.14H.sub.17N.sub.3OS, 275.11; m/z found, 276.2 [M+H].sup.+.
.sup.1H NMR (CDCl.sub.3, 400 MHz): 11.1 (br s, 1H), 6.96 (d, J=5.4
Hz, 1H), 6.87 (d, J=5.4 Hz, 1H), 6.71 (d, J=1.9 Hz, 1H), 4.84 (d,
J=12.2 Hz, 1H), 4.70 (d, J=12.2 Hz, 1H), 3.30-2.90 (m, 4H),
2.30-1.40 (m, 7H).
Example 3
[0114] ##STR12##
(2-Chloro-6H-thieno[2,3-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-methanone
[0115] A. 2-Chloro-6H-thieno[2,3-b]pyrrole-5-carboxylic acid ethyl
ester. A solution of 6H-thieno[2,3-b]pyrrole-5-carboxylic acid
ethyl ester (580 mg, 3.0 mmol) in acetic acid (6 mL) and CHCl.sub.3
(6 mL) was treated with three portions of N-chlorosuccinimide
(total 415 mg, 3.15 mmol) at 0.degree. C. over 2 h. The reaction
mixture was slowly warmed to room temperature and stirred
overnight. The CHCl.sub.3 was then removed, and the residue was
basified with 4 N NaOH and extracted with EtOAc. The combined
organics were washed with satd aq NaHCO.sub.3, dried over
Na.sub.2SO.sub.4, and concentrated. Column chromatography
(SiO.sub.2, 5-10% EtOAc/hexanes) gave 600 mg (88%) of a white
solid. TLC (silica, 20% EtOAc/hexanes): R.sub.f=0.5. .sup.1H NMR
(CDCl.sub.3, 400 MHz): 10.5 (br s, 1H), 6.97 (d, J=2.0 Hz, 1H),
6.85 (s, 1H), 4.39 (q, J=7.2 Hz, 2H), 1.35 (t, J=7.2 Hz, 3H).
[0116] B.
(2-Chloro-6H-thieno[2,3-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl-
)-methanone.
[0117] 2-Chloro-6H-thieno[2,3-b]pyrrole-5-carboxylic acid ethyl
ester (102 mg, 0.45 mmol) was hydrolyzed (Procedure B) and coupled
with N-methylpiperazine (procedure D) to provide the title compound
(102 mg, 80% for two steps) as an off-white solid. TLC (silica, 10%
MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.4. MS (electrospray): exact mass
calculated for C.sub.12H.sub.14ClN.sub.3OS, 283.05; m/z found,
284.1 [M+H].sup.+. .sup.1H NMR (CDCl.sub.3, 400 MHz): 10.5 (br s,
1H), 6.87 (s, 1H), 6.61 (d, J=1.8 Hz, 1H), 3.92 (t, J=5.1 Hz, 4H),
2.50 (t, J=5.1 Hz, 4H), 2.35 (s, 3H).
Example 4
[0118] ##STR13##
(2-Chloro-6H-thieno[2,3-b]pyrrol-5-yl)-(hexahydro-pyrrolo[1,2-a]pyrazin-2--
yl)-methanone
[0119] 2-Chloro-6H-thieno[2,3-b]pyrrole-5-carboxylic acid ethyl
ester (102 mg, 0.45 mmol) was hydrolyzed (Procedure B) and then
coupled with octahydro-pyrrolo[1,2-a]pyrazine (Procedure D) to
provide the title compound (108 mg, 78% for two steps) as an
off-white solid. TLC (silica, 10% MeOH/CH.sub.2Cl.sub.2):
R.sub.f=0.35. MS (electrospray): exact mass calculated for
C.sub.14H.sub.16ClN.sub.3OS, 309.07; m/z found, 310.1 [M+H].sup.+.
.sup.1H NMR (CDCl.sub.3, 400 MHz): 11.1 (br s, 1H), 6.86 (s, 1H),
6.62 (s, 1H), 4.79 (d, J=11.8 Hz, 1H), 4.67 (d, J=11.8 Hz, 1H),
3.30-2.90 (m, 4H), 2.30-1.40 (m, 7H).
Example 5
[0120] ##STR14##
(2-Chloro-6H-thieno[2,3-b]pyrrol-5-yl)-piperazin-1-yl-methanone
[0121] 2-Chloro-6H-thieno[2,3-b]pyrrole-5-carboxylic acid ethyl
ester (102 mg, 0.45 mmol) was hydrolyzed (Procedure B) and then
coupled with piperazine (Procedure D) to provide the title compound
(42 mg, 35% for two steps) as an off-white solid. TLC (silica, 10%
MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.15. MS (electrospray): exact mass
calculated for C.sub.11H.sub.12ClN.sub.3OS, 269.04; m/z found,
270.1 [M+H].sup.+. .sup.1H NMR (CDCl.sub.3, 400 MHz): 10.5 (br s,
1H), 6.87 (s, 1H), 6.61 (s, 1H), 3.87 (t, J=4.8 Hz, 4H), 2.96 (t,
J=5.2 Hz, 4H).
Example 6
[0122] ##STR15##
(4H-Furo[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-methanone
[0123] A. 4H-Furo[3,2-b]pyrrole-5-carboxylic acid ethyl ester.
Furan-2-carbaldehyde (1.92 g, 20 mmol) was annulated according to
procedure A to provide the title compound (1.97 g, 55%) as a white
solid. TLC (silica, 20% EtOAc/hexanes): R.sub.f=0.50. .sup.1H NMR
(CDCl.sub.3, 400 MHz): 8.95 (br s, 1H), 7.51 (d, J=2.2 Hz, 1H),
6.81-6.80 (m, 1H), 6.46-6.45 (m, 1H), 4.35 (q, J=7.1 Hz, 2H), 1.38
(t, J=7.1 Hz, 3H).
[0124] B.
(4H-Furo[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-methanone- .
4H-Furo[3,2-b]pyrrole-5-carboxylic acid ethyl ester (200 mg, 1.12
mmol) was hydrolyzed (Procedure B) and coupled with
N-methylpiperazine (Procedure D) to provide the title compound (185
mg, 71% for two steps) as an off-white solid. TLC (silica, 10%
MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.4. MS (electrospray): exact mass
calculated for C.sub.12H.sub.15N.sub.3O.sub.2, 233.12; m/z found,
234.2 [M+H].sup.+. .sup.1H NMR (CDCl.sub.3, 400 MHz): 10.3 (br s,
1H), 7.43 (d, J=2.2 Hz, 1H), 6.43-6.42 (m, 2H), 3.90 (t, J=5.0 Hz,
4H), 2.47 (t, J=5.1 Hz, 4H), 2.32 (s, 3H).
Example 7
[0125] ##STR16##
(4-Methyl-piperazin-1-yl)-(4H-thieno[3,2-b]pyrrol-5-yl)-methanone
[0126] A. 4H-Thieno[3,2-b]pyrrole-5-carboxylic acid ethyl ester. To
a solution of thiophene-2-carbaldehyde (1.10 mL, 11.7 mmol) and
ethyl azidoacetate (1.4 mL, 11.7 mmol) in EtOH (35 mL) cooled to
0.degree. C. was added NaOEt (1.0 g, 14.7 mmol) in one portion. The
mixture was allowed to reach room temperature over 14 h and was
then poured into water (400 mL) and extracted with CH.sub.2Cl.sub.2
(3.times.50 mL). The combined organics were washed with water and
brine, dried over Na.sub.2SO.sub.4, and concentrated. The residue
was taken up in xylenes (10 mL), and the resulting solution was
refluxed for 1 h. The solution was cooled and then loaded directly
onto silica gel and purified (35 g SiO.sub.2, 10-20% EtOAc/hexanes)
to reveal 0.12 g (5%) of a yellowish solid. .sup.1H NMR (400 MHz,
CDCl.sub.3): 9.06 (br s, 1H), 7.33 (d, J=5.3 Hz, 1H), 7.15-7.14 (m,
1H), 6.96 (dd, J=5.3, 0.8 Hz, 1H), 4.37 (q, J=7.3 Hz, 2H), 1.39 (t,
J=7.3 Hz, 3H). .sup.13C NMR (100 MHz, CDCl.sub.3): 161.3, 140.9,
129.2, 126.9, 124.6, 110.9, 107.3, 60.4, 14.2.
[0127] B.
(4-Methyl-piperazin-1-yl)-(4H-thieno[3,2-b]pyrrol-5-yl)-methano-
ne. To a solution of 4H-thieno[3,2-b]pyrrole-5-carboxylic acid
ethyl ester (98.5 mg, 0.50 mmol) in wet THF (3 mL) was added LiOH
(129 mg, 3 mmol). This mixture was stirred at room temperature for
3 days. The reaction mixture was diluted with water (50 mL), and 1
M HCL was added to adjust the pH to about 3. This mixture was then
extracted with EtOAc, and the combined organics were dried over
Na.sub.2SO.sub.4. The solvent was removed to reveal 73.4 mg (87%)
of the free acid, which was used in the coupling event without
further purification. The acid (73.4 mg, 0.44 mmol) was taken up in
THF (3 mL), and CDI (87.1 mg, 0.54 mmol) was added in one portion.
The reaction mixture was stirred for 1 h. To this mixture was then
added 1-methylpiperazine (70 .mu.L), and the mixture stirred for an
additional 6 h. The reaction mixture was diluted with EtOAc, washed
with water, NaHCO.sub.3 (aq) and then brine, and subsequently
purified by column chromatography (10 g SiO.sub.2, 1-8% MeOH (2 M
NH.sub.3)/CH.sub.2Cl.sub.2) to reveal 55.6 mg (51%) of the title
compound. .sup.1H NMR (400 MHz, CDCl.sub.3): 9.26 (br s, 1H), 7.26
(d, J=5.3 Hz, 1H), 6.97 (dd, J=5.3, 0.8 Hz, 1H), 6.75-6.74 (m, 1H),
4.07-3.88 (m, 4H), 2.68-2.48 (m, 4H), 2.43 (br s, 3H). MS
(electrospray): exact mass calculated for
C.sub.12H.sub.15N.sub.3OS, 249.09; m/z found, 250.1
[M+H].sup.+.
Example 8
[0128] ##STR17##
Piperazin-1-yl-(4H-thieno[3,2-b]pyrrol-5-yl)-methanone
[0129] 4H-Thieno[3,2-b]pyrrole-5-carboxylic acid (50 mg, 0.30 mmol)
was coupled with piperazine according to Procedure D to provide the
title compound (25 mg, 35%) as an off-white solid. TLC (silica, 10%
MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.15. MS (electrospray): exact mass
calculated for C.sub.11H.sub.13N.sub.3OS, 235.08; m/z found, 236.1
[M+H].sup.+. .sup.1H NMR (CD.sub.3OD, 400 MHz): 7.33 (d, J=5.3 Hz,
1H), 6.98 (dd, J=5.2, 0.7 Hz, 1H), 6.89 (d, J=0.6 Hz, 1H), 4.08 (t,
J=5.3 Hz, 4H), 3.50-3.20 (m, 4H).
Example 9
[0130] ##STR18##
(3-Methyl-piperazin-1-yl)-(4H-thieno[3,2-b]pyrrol-5-yl)-methanone
[0131] 4H-Thieno[3,2-b]pyrrole-5-carboxylic acid (50 mg, 0.30 mmol)
was coupled with 2-methylpiperazine according to Procedure D to
provide the title compound (58 mg, 78%) as an off-white solid. TLC
(silica, 10% MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.15. MS
(electrospray): exact mass calculated for
C.sub.12H.sub.15N.sub.3OS, 249.09; m/z found, 250.1 [M+H].sup.+.
.sup.1H NMR (CD.sub.3OD, 400 MHz): 7.33 (d, J=5.3 Hz, 1H), 6.98 (d,
J=5.3 Hz, 1H), 6.88 (s, 1H), 4.62-4.56 (m, 2H), 3.50-3.20 (m, 5H),
1.36 (d, J=6.6 Hz, 3H).
Example 10
[0132] ##STR19##
(2-Chloro-4H-thieno[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-methanone
[0133] A. 2-Chloro-4H-thieno[3,2-b]pyrrole-5-carboxylic acid ethyl
ester. 5-Chloro-thiophene-2-carbaldehyde (2.92 g, 20 mmol) was
annulated according to Procedure A to provide the title compound
(2.8 g, 61%) as a white solid. TLC (silica, 20% EtOAc/hexanes):
R.sub.f=0.48. .sup.1H NMR (CDCl.sub.3, 400 MHz): 9.10 (br s, 1H),
7.04 (dd, J=1.9, 0.7 Hz, 1H), 6.89 (d, J=0.7 Hz, 1H), 4.37 (q,
J=7.2 Hz, 2H), 1.39 (t, J=7.2 Hz, 3H).
[0134] B.
(2-Chloro-4H-thieno[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl-
)-methanone.
[0135] 2-Chloro-4H-thieno[3,2-b]pyrrole-5-carboxylic acid ethyl
ester (230 mg, 1.0 mmol) was hydrolyzed (Procedure B) and then
coupled with N-methylpiperazine (Procedure C) to provide the title
compound (128 mg, 45% for two steps) as an off-white solid. TLC
(silica, 10% MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.4. MS
(electrospray): exact mass calculated for
C.sub.12H.sub.14ClN.sub.3OS, 283.05; m/z found, 284.1 [M+H].sup.+.
.sup.1H NMR (CDCl.sub.3, 400 MHz): 10.1 (br s, 1H), 6.88 (s, 1H),
6.64 (d, J=1.4 Hz, 1H), 3.91 (t, J=4.4 Hz, 4H), 2.49 (t, J=5.1 Hz,
4H), 2.35 (s, 3H).
Example 11
[0136] ##STR20##
(2-Chloro-4H-thieno[3,2-b]pyrrol-5-yl)-(hexahydro-pyrrolo[1,2-a]pyrazin-2--
yl)-methanone
[0137] 2-Chloro-4H-thieno[3,2-b]pyrrole-5-carboxylic acid ethyl
ester (230 mg, 1.0 mmol) was hydrolyzed (Procedure B) and then
coupled with octahydro-pyrrolo[1,2-a]pyrazine (Procedure C) to
provide the title compound (93 mg, 30% for two steps) as an
off-white solid. TLC (silica, 10% MeOH/CH.sub.2Cl.sub.2):
R.sub.f=0.4. MS (electrospray): exact mass calculated for
C.sub.14H.sub.16ClN.sub.3OS, 309.07; m/z found, 310.1 [M+H].sup.+.
.sup.1H NMR (CDCl.sub.3, 400 MHz): 10.9 (br s, 1H), 6.86 (s, 1H),
6.64 (d, J=1.4 Hz, 1H), 4.77 (d, J=12.2 Hz, 1H), 4.65 (d, J=12.7
Hz, 1H), 3.30-2.90 (m, 4H), 2.30-1.40 (m, 7H).
Example 12
[0138] ##STR21##
[0139]
(3-Bromo-4H-thieno[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-me-
thanone.
[0140] A. 3-Bromo-4H-thieno[3,2-b]pyrrole-5-carboxylic acid ethyl
ester. 4-Bromo-thiophene-2-carbaldehyde (3.8 g, 20 mmol) was
annulated according to Procedure A to provide the title compound
(1.2 g, 22%) as a white solid. TLC (silica, 20% EtOAc/hexanes):
R.sub.f=0.48. .sup.1H NMR (CDCl.sub.3, 400 MHz): 9.58 (br s, 1H),
7.20 (s, 1H), 7.15 (d, J=1.5 Hz, 1H), 4.41 (q, J=7.2 Hz, 2H), 1.39
(t, J=7.2 Hz, 3H).
[0141] B.
(3-Bromo-4H-thieno[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-
-methanone.
[0142] 3-Bromo-4H-thieno[3,2-b]pyrrole-5-carboxylic acid ethyl
ester (67 mg, 0.24 mmol) was hydrolyzed (Procedure B) and then
coupled with N-methylpiperazine (Procedure D) to provide the title
compound (65 mg, 82% for two steps) as an off-white solid. TLC
(silica, 10% MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.4. MS
(electrospray): exact mass calculated for
C.sub.12H.sub.14BrN.sub.3OS, 327.00; m/z found, 328.0 [M+H].sup.+.
.sup.1H NMR (CDCl.sub.3, 400 MHz): 9.95 (br s, 1H), 7.11 (s, 1H),
6.73 (d, J=1.8 Hz, 1H), 3.91 (t, J=5.1 Hz, 4H), 2.49 (t, J=5.1 Hz,
4H), 2.34 (s, 3H).
Example 13
[0143] ##STR22##
(4-Methyl-piperazin-1-yl)-(3-methyl-4H-thieno[3,2-b]pyrrol-5-yl)-methanone
[0144] A. 4-methyl-thiophene-2-carbaldehyde. A solution of
3-methylthiophene (6.76 mL, 70 mmol) in ether (70 mL) was treated
with n-butyllithium (2.5 M in hexanes, 28.6 mL, 71.4 mmol) at such
a rate that a slight reflux was maintained. The reaction mixture
was heated to reflux for 15 min and then DMF (7.0 mL, 91 mmol) in
ether (30 mL) was added. After stirring for 4 h, the reaction was
quenched with addition of satd aq NH.sub.4Cl (200 mL). The organic
layer was separated, washed with brine and then H.sub.2O, dried
over Na.sub.2SO.sub.4, and concentrated. Column chromatography
(SiO.sub.2, 5-10% EtOAc/hexanes) provided a mixture of
4-methyl-thiophene-2-carbaldehyde and
3-methyl-thiophene-2-carbaldehyde (4.4:1, 8.1 g, 92%) as a light
yellow oil. TLC (silica, 10% EtOAc/hexanes): R.sub.f=0.55. For
4-methyl-thiophene-2-carbaldehyde: .sup.1H NMR (CDCl.sub.3, 400
MHz): 9.95 (s, 1H), 7.58 (d, J=1.2 Hz, 1H), 7.37-7.35 (m, 1H), 2.32
(s, 3H). For 3-methyl-thiophene-2-carbaldehyde: .sup.1H NMR
(CDCl.sub.3, 400 MHz): 10.02 (s, 1H), 7.64 (d, J=4.6 Hz, 1H), 6.97
(d, J=4.6 Hz, 1H), 2.58 (s, 3H).
[0145] B. 3-Methyl-4H-thieno[3,2-b]pyrrole-5-carboxylic acid ethyl
ester. The mixture of 4-methyl-thiophene-2-carbaldehyde and
3-methyl-thiophene-2-carbaldehyde (2.84 g, 22.5 mmol) was annulated
according to Procedure A to provide the title compound (2.5 g, 65%)
as a white solid. TLC (silica, 20% EtOAc/hexanes): R.sub.f=0.45.
.sup.1H NMR (CDCl.sub.3, 400 MHz): 9.95 (br s, 1H), 7.12 (d, J=1.9
Hz, 1H), 6.90 (d, J=1.2 Hz, 1H), 4.39 (q, J=7.2 Hz, 2H), 2.35 (s,
3H), 1.39 (t, J=7.2 Hz, 3H).
[0146] C.
(4-Methyl-piperazin-1-yl)-(3-methyl-4H-thieno[3,2-b]pyrrol-5-yl-
)-methanone. 3-Methyl-4H-thieno[3,2-b]pyrrole-5-carboxylic acid
ethyl ester (200 mg, 0.96 mmol) was hydrolyzed (Procedure B) and
then coupled with N-methylpiperazine (Procedure D) to provide the
title compound (197 mg, 78% for two steps) as an off-white solid.
TLC (silica, 10% MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.4. MS
(electrospray): exact mass calculated for
C.sub.13H.sub.17N.sub.3OS, 263.11; m/z found, 264.1 [M+H].sup.+.
.sup.1H NMR (CDCl.sub.3, 400 MHz): 11.10 (br s, 1H), 6.76 (d, J=1.2
Hz, 1H), 6.69 (d, J=2.0 Hz, 1H), 3.94-3.90 (m, 4H), 2.47 (t, J=5.1
Hz, 4H), 2.33 (s, 3H), 2.25 (s, 3H).
Example 14
[0147] ##STR23##
(2-Methyl-4H-furo[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-methanone
[0148] A. 2-Methyl-4H-furo[3,2-b]pyrrole-5-carboxylic acid ethyl
ester. 5-Methyl-furan-2-carbaldehyde (2.2 g, 20 mmol) was annulated
according to Procedure A to provide the title compound (2.89 g,
75%) as a white solid. TLC (silica, 10% EtOAc/hexanes):
R.sub.f=0.4. .sup.1H NMR (CDCl.sub.3, 400 MHz): 9.50 (br s, 1H),
6.73 (s, 1H), 6.04 (s, 1H), 4.35 (q, J=7.2 Hz, 2H), 2.37 (s, 3H),
1.35 (t, J=7.2 Hz, 3H).
[0149] B.
(2-Methyl-4H-furo[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)--
methanone. 2-Methyl-4H-furo[3,2-b]pyrrole-5-carboxylic acid ethyl
ester (200 mg, 1.04 mmol) was hydrolyzed (Procedure B) and then
coupled with N-methylpiperazine (Procedure D) to provide the title
compound (208 mg, 81% for two steps) as a white solid. TLC (silica,
10% MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.35. MS (electrospray): exact
mass calculated for C.sub.13H.sub.17N.sub.3O.sub.2, 247.13; m/z
found, 248.2 [M+H].sup.+. .sup.1H NMR (CDCl.sub.3, 400 MHz): 9.85
(br s, 1H), 6.36 (s, 1H), 6.07 (s, 1H), 3.87 (t, J=5.0 Hz, 4H),
2.47 (t, J=5.2 Hz, 4H), 2.39 (s, 3H), 2.33 (s, 3H).
Example 15
[0150] ##STR24##
(2,3-Dimethyl-4H-furo[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-methano-
ne
[0151] A. 2,3-Dimethyl-4H-furo[3,2-b]pyrrole-5-carboxylic acid
ethyl ester. 4,5-Dimethyl-furan-2-carbaldehyde (2.2 g, 18 mmol) was
annulated according to Procedure A to provide the title compound
(1.76 g, 48%) as an off-white solid. TLC (silica, 10%
EtOAc/hexanes): R.sub.f=0.35. .sup.1H NMR (CDCl.sub.3, 400 MHz):
8.95 (br s, 1H), 6.69 (d, J=1.7 Hz, 1H), 4.32 (q, J=7.2 Hz, 2H),
2.33 (s, 3H), 2.08 (s, 3H), 1.37 (t, J=7.2 Hz, 3H).
[0152] B.
(2,3-Dimethyl-4H-furo[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1--
yl)-methanone. 2,3-Dimethyl-4H-furo[3,2-b]pyrrole-5-carboxylic acid
ethyl ester (200 mg, 0.97 mmol) was hydrolyzed (Procedure B) and
then coupled with N-methylpiperazine (Procedure D) to provide the
title compound (190 mg, 75% for two steps) as an off-white solid.
TLC (silica, 10% MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.35. MS
(electrospray): exact mass calculated for
C.sub.14H.sub.19N.sub.3O.sub.2, 261.15; m/z found, 261.8
[M+H].sup.+. .sup.1H NMR (CDCl.sub.3, 400 MHz): 9.95 (br s, 1H),
6.32 (d, J=1.8 Hz, 1H), 3.88 (t, J=5.0 Hz, 4H), 2.47 (t, J=5.1 Hz,
4H), 2.33 (s, 3H), 2.31 (s, 3H), 2.06 (s, 3H).
Example 16
[0153] ##STR25##
(2,3-Dimethyl-4H-thieno[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-metha-
none
[0154] A. 2,3-Dimethyl-4H-thieno[3,2-b]pyrrole-5-carboxylic acid
ethyl ester. 4,5-Dimethyl-thiophene-2-carbaldehyde (2.0 g, 14 mmol)
was annulated according to Procedure A to provide the title
compound (160 mg, 5%) as a white solid. TLC (silica, 10%
EtOAc/hexanes): R.sub.f=0.40. .sup.1H NMR (CDCl.sub.3, 400 MHz):
9.50 (br s, 1H), 7.05 (d, J=1.9 Hz, 1H), 4.36 (q, J=7.2 Hz, 2H),
2.41 (s, 3H), 2.22), (s, 3H), 1.38 (t, J=7.2 Hz, 3H).
[0155] B.
(2,3-Dimethyl-4H-thieno[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin--
1-yl)-methanone. 2,3-Dimethyl-4H-thieno[3,2-b]pyrrole-5-carboxylic
acid ethyl ester (68 mg, 0.30 mmol) was hydrolyzed (Procedure B)
and then coupled with N-methylpiperazine (Procedure D) to provide
the title compound (67 mg, 80% for two steps) as an off-white
solid. TLC (silica, 10% MeOH/CH.sub.2Cl.sub.2): R.sub.f=0.35. MS
(electrospray): exact mass calculated for
C.sub.14H.sub.19N.sub.3OS, 277.12; m/z found, 278.1 [M+H].sup.+.
.sup.1H NMR (CDCl.sub.3, 400 MHz): 10.95 (br s, 1H), 6.63 (d, J=1.9
Hz, 1H), 3.92 (t, J=4.5 Hz, 4H), 2.46 (t, J=5.0 Hz, 4H), 2.36 (s,
3H), 2.33 (s, 3H), 2.13 (s, 3H).
Examples 17-25
[0156] The following compounds were made according to the synthetic
methods outlined in Schemes 1-4:
EX Compound
[0157] 17
(2,3-Dichloro-6H-thieno[2,3-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-meth-
anone; [0158] 18
(2-Methyl-4H-furo[3,2-b]pyrrol-5-yl)-piperazin-1-yl-methanone;
[0159] 19
(3-Bromo-4H-thieno[3,2-b]pyrrol-5-yl)-piperazin-1-yl-methanone;
[0160] 20
(3-Bromo-4H-thieno[3,2-b]pyrrol-5-yl)-(3-methyl-piperazin-1-yl)-methanone-
; [0161] 21
(3-Methyl-4H-thieno[3,2-b]pyrrol-5-yl)-piperazin-1-yl-methanone;
[0162] 22
(3-Methyl-piperazin-1-yl)-(3-methyl-4H-thieno[3,2-b]pyrrol-5-yl)-metha-
none; [0163] 23
(2-Chloro-3-methyl-4H-thieno[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-
-methanone; [0164] 24
(2-Chloro-3-methyl-4H-thieno[3,2-b]pyrrol-5-yl)-piperazin-1-yl-methanone;
and [0165] 25
(2,3-Dichloro-4H-thieno[3,2-b]pyrrol-5-yl)-(4-methyl-piperazin-1-yl)-meth-
anone.
Biological Examples
[0165] Binding Assay on Recombinant Human Histamine H.sub.4
Receptor
[0166] SK-N-MC cells or COS7 cells were transiently transfected
with pH4R and grown in 150 cm.sup.2 tissue culture dishes. Cells
were washed with saline solution, scraped with a cell scraper and
collected by centrifugation (1000 rpm, 5 min). Cell membranes were
prepared by homogenization of the cell pellet in 20 mM Tris-HCl
with a polytron tissue homogenizer for 10 s at high speed.
Homogenate was centrifuged at 1000 rpm for 5 min at 4.degree. C.
The supernatant was then collected and centrifuged at
20,000.times.g for 25 min at 4.degree. C. The final pellet was
resuspended in 50 mM Tris-HCl. Cell membranes were incubated with
.sup.3H-histamine (5-70 nM) in the presence or absence of excess
histamine (10000 nM). Incubation occurred at room temperature for
45 min. Membranes were harvested by rapid filtration over Whatman
GF/C filters and washed 4 times with ice-cold 50 mM Tris HCl.
Filters were then dried, mixed with scintillant and counted for
radioactivity. SK-N-MC or COS7 cells expressing human histamine
H.sub.4 receptor were used to measure the affinity of binding of
other compounds and their ability to displace .sup.3H-ligand
binding by incubating the above-described reaction in the presence
of various concentrations of inhibitor or compound to be tested.
For competition binding studies using .sup.3H-histamine, K.sub.i
values were calculated, based on an experimentally determined
K.sub.D value of 5 nM and a ligand concentration of 5 nM, according
to Y.-C. Cheng and W. H. Prusoff (Biochem. Pharmacol. 1973,
22(23):3099-3108): K.sub.i=(IC.sub.50)/(1+([L]/(K.sub.D)).
TABLE-US-00001 BINDING ASSAY RESULTS EX K.sub.i (nM) 1 85 2 461 3
25 4 176 5 56 6 840 7 125 8 343 9 733 10 40 11 715 12 56 13 21 14
343 15 140 16 5 17 10 18 770 19 410 20 980 21 80 22 161 23 3 24 30
25 5.5
Mast Cell Chemotaxis Assay
[0167] Mast cell accumulation in mucosal epithelia is a well-known
characteristic of allergic rhinitis and asthma. Transwells (Costar,
Cambridge, Mass.) of a pore size 8 .mu.m were coated with 100 .mu.L
of 100 ng/mL human fibronectin (Sigma) for 2 h at room temperature.
After removal of the fibronectin, 600 .mu.L of RPMI with 5% BSA, in
the presence of 10 .mu.M histamine, was added to the bottom
chamber. To test the various histamine receptor (HR) antagonists,
10 .mu.M and/or 1 .mu.M Solutions of the test compounds were added
to the top and bottom chambers. Mast cells (2.times.10.sup.5/well)
were added to the top chamber. The plates were Incubated for 3 h at
37.degree. C. Transwells were removed and the cells in the Bottom
chamber were counted for sixty seconds using a flow cytometer.
TABLE-US-00002 10 .mu.M Histamine HR Antagonist (.mu.M): Binding 10
1 Assay EX % Inh Stdev % Inh Stdev K.sub.i (nM) 3 106 4 103 0 25 4
<5 -- <5 -- 176 10 92 3 40 13 60 20 21 20 <5 -- <5 --
980
Cell-Type Distribution of H.sub.4 Expression
[0168] RNA was prepared from the different cells using an RNeasy
kit (Qiagen, Valencia, Calif.) according to the manufacturer's
instructions. RNA samples (5 .mu.g) were run on an RNA gel and then
transferred overnight to a nylon blot (Hybond, Amersham Pharmacia
Biotech, Piscataway, N.J.). The blot was pre-hybridized with
ExpressHyb solution (CLONTECH) for 30 min at 68.degree. C. The
H.sub.4 receptor DNA was labeled using the Rediprime II kit
(Amersham Pharmacia Biotech). The blot was hybridized for 2 h at
68.degree. C., followed by one wash step (23 SSC and 0.05% SDS) of
40 min at room temperature, and a second wash step (0.13 SSC and
0.1% SDS) of 40 min at 50.degree. C. The blot was exposed to X-ray
film at -70.degree. C. with two intensifying screens overnight.
Results
[0169] The Northern Blot results indicate that the H.sub.4 receptor
is expressed on bone marrow-derived mast cells (BMMC), peritoneal
mast cells, and eosinophils. These positive results are consistent
with the published literature (e.g. Oda et al., Nguyen et al., and
Morse et al. in the Background section). However, the negative
results of the Northern Blot experiment, such as the finding of
apparently no measurable levels of H.sub.4 receptor expressed by
neutrophils, differ somewhat from the above literature findings.
This may be explained by the different methodologies used.
Accumulation of mast cells and eosinophils in affected tissues is
one of the principal characteristics of allergic rhinitis and
asthma. Since H.sub.4 receptor expression is limited to these cell
types; H.sub.4 receptor signalling is likely to mediate the
infiltration of mast cells and eosinophils in response to
histamine. Additional investigation may also clarify these issues.
The following table reports the Cell-type Distribution of H.sub.4
Expression by Northern Blot. TABLE-US-00003 Species Cell Type
H.sub.4 Human Eosinophils + Immature Dendritic Cells - Mature
Dendritic Cells - CD14.sup.+ Monocytes - CD4.sup.+ T Cells -
CD8.sup.+ T Cells - B Cells - Neutrophils - Mouse/(Rat) Eosinophils
+ Peritoneal Mast Cells (Rat) + BMMC + BM Derived Macrophages -
Peritoneal Macrophages - CD4.sup.+ T Cells - B Cells -
The Inhibition of Eosinophil Shape Change by Histamine H.sub.4
Receptor Antagonists
[0170] Eosinophil accumulation in sites of allergic reaction is a
well-known characteristic of allergic rhinitis and asthma. This
example demonstrates that histamine H.sub.4 receptor antagonists
can block the shape change response in human eosinophils in
response to histamine. Shape change is a cellular characteristic
that precedes eosinophil chemotaxis.
Methods
[0171] Human granulocytes were isolated from human blood by a
Ficoll gradient. The red blood cells were lysed with 5-10.times.
Qiagen lysis buffer at room temperature for 5-7 min. Granulocytes
were harvested and washed once with FACS buffer. The cells were
resuspended at a density of 2.times.10.sup.6 cells/mL in reaction
buffer. To test inhibition by specific histamine receptor
antagonists, 90 .mu.L of the cell suspension
(.about.2.times.10.sup.5 cells) was incubated with 10 .mu.M of one
of the various test compound solutions. After 30 min, 11 .mu.L of
one of the various concentrations of histamine was added. Ten
minutes later the cells were transferred to ice and fixed with 250
.mu.L of ice-cold fixative buffer (2% formaldehyde) for 1 min. The
shape change was quantitated using a gated autofluorescence forward
scatter assay (GAFS) (Byran et al., Am. J. Crit. Care Med.
2002,165:1602-1609).
Results--Histamine Mediates Eosinophil Shape Change Through H.sub.4
Receptor
[0172] The change in shape of eosinophils is due to cytoskeletal
changes that preceed chemotaxis and thus is a measure of
chemotaxis. The data in the following table show that histamine
induces a dose-dependent shape change in eosinophils. Histamine
receptor (HR) antagonists were used to sort out which histamine
receptor is responsible for the shape change. Antagonists specific
for the histamine H.sub.1 receptor (diphenhydramine) or the H.sub.2
receptor (ranatidine) did not alter the histamine-induced shape
change. However, a dual H.sub.3/H.sub.4 antagonist (thioperamide)
and a specific histamine H.sub.4 receptor antagonist
((5-Chloro-1H-indol-2-yl)-(4-methyl-piperazin-1-yl)-methanone,
K.sub.i=5 nM) inhibited histamine-induced eosinophil shape change
with an IC.sub.50 of 1.5 and 0.27 .mu.M, respectively.
TABLE-US-00004 Histamine Fold Change (.mu.M): 10 1 0.1 0.01 0 No HR
1.34 1.31 1.21 1.01 1.00 Antagonist 10 .mu.M H.sub.4 1.09 1.05 1.05
1.01 1.00 Antagonist 10 .mu.M 1.08 1.05 1.01 1.04 1.00 Thiop 10
.mu.M 1.63 1.50 1.18 1.03 1.00 Diphen 10 .mu.M 1.64 1.49 1.21 1.04
1.00 Ranat
The Inhibition of Eosinophil Chemotaxis by Histamine H.sub.4
Receptor Antagonists
[0173] Eosinophil accumulation in sites of allergic reaction is a
well-known characteristic of allergic rhinitis and asthma.
Eosinophils are purified from human blood with standard methods.
Chemotaxis assays are carried out using transwells (Costar,
Cambridge, Mass.) of a pore size 5 .mu.m coated with 100 .mu.L of
100 ng/mL human fibronectin (Sigma) for 2 h at room temperature.
After removal of the fibronectin, 600 .mu.L of RPMI with 5% BSA in
the presence of histamine (ranging from 1.25-20 .mu.M) is added to
the bottom chamber. To test the various histamine receptor
antagonists 10 .mu.M of the test compounds can be added to the top
and bottom chambers. Eosinophils will be added to the top chamber
whereas histamine or chemotactic factors will be placed in the
lower chamber. The plates are incubated for 3 h at 37.degree. C.
Transwells are removed and the number of cells in the bottom
chamber can be counted for 60 s using a flow cytometer, or can be
quantitated by using Giemsa staining.
The Inhibition of Zymosan-Induced Peritonitis in Mice by Histamine
H.sub.4 Receptor Antagonists
[0174] It has been demonstrated that histamine H.sub.4 receptor
antagonists can block the peritonitis induced by zymosan, which is
the insoluble polysaccharide component on the cell wall of
Saccharomyces cerevisiae. This is commonly used to induce
peritonitis in mice and appears to act in a mast cell-dependent
manner. Compounds of the present invention can be tested in such a
model to demonstrate their use as anti-inflammatory agents. At time
0 mice are given compound or PBS, either s.c. or p.o. Fifteen
minutes later each mouse receives 1 mg zymosan A (Sigma) i.p. The
mice are sacrificed 4 h later, and the peritoneal cavities are
washed with 3 mL of PBS containing 3 mM EDTA. The number of
migrated leukocytes is determined by taking an aliquot (100 .mu.L)
of the lavage fluid and diluting 1:10 in Turk's solution (0.01%
crystal violet in 3% acetic acid). The samples are then vortexed,
and 10 .mu.L of the stained cell solution is placed in a Neubauer
haemocytometer. Differential cell counts are performed using a
light microscope (Olympus B061). In view of their chromatic
characteristics and their nucleus and cytoplasm appearance,
polymorphonuclear leukocytes (PMN; >95% neutrophils) can be
easily identified. Treatment with zymosan increases the number of
neutrophils, which is representative of an inflammatory response.
Treatment with H.sub.4 receptor antagonist will block this
increase.
Inhibition of Mast Cell Chemotaxis by H.sub.4 Receptor Antagonist
in an Animal Model of Asthma and Allergic Rhinitis
[0175] An animal model will be used to test the observation that
mast cells accumulate in response to allergic inflammation and that
this can be blocked by H.sub.4 receptor antagonists. Compounds of
the present invention can be tested in this model to demonstrate
their use as treatments for allergic rhinitis or asthma. Mice will
be sensitized by intraperitoneal injection of ovalbumin/Alum (10
.mu.g in 0.2 ml Al(OH).sub.3; 2%) on Day 0 and Day 14. On Day 21
through 23 mice will be challenged by PBS or ovalbumin, and
sacrificed 24 h after the last challenge on Day 24. A section of
the trachea will be removed and fixed in formalin. Paraffin
embedding and longitudinal sectioning of tracheas will be performed
followed by staining of mast cells with toluidine blue.
Alternatively, trachea will be frozen in OCT for frozen sectioning,
and mast cells will be identified by IgE staining. Mast cells will
be quantified as sub-mucosal or sub-epithelial depending on their
location within each tracheal section. Exposure to allergen should
increase the number of sub-epithelial mast cells, and this effect
will be blocked by H.sub.4 receptor antagonists.
[0176] The features and advantages of the invention are apparent to
one of ordinary skill in the art. Based on this disclosure,
including the summary, detailed description, background, examples,
and claims, one of ordinary skill in the art will be able to make
modifications and adaptations to various conditions and usages.
Publications described herein are incorporated by reference in
their entirety. These other embodiments are also within the scope
of the invention.
* * * * *