U.S. patent application number 15/408896 was filed with the patent office on 2017-07-20 for 2-oxindole compounds.
The applicant listed for this patent is CHEMOCENTRYX, INC.. Invention is credited to Xi CHEN, Dean R. DRAGOLI, Pingchen FAN, Manmohan Reddy LELETI, Rebecca M. LUI, Viengkham MALATHONG, Jay P. POWERS, Rajinder SINGH, Hiroko TANAKA, Ju YANG, Chao YU, Penglie ZHANG.
Application Number | 20170204087 15/408896 |
Document ID | / |
Family ID | 59314376 |
Filed Date | 2017-07-20 |
United States Patent
Application |
20170204087 |
Kind Code |
A1 |
CHEN; Xi ; et al. |
July 20, 2017 |
2-OXINDOLE COMPOUNDS
Abstract
Oxindole compounds useful for the treatment of CCR(9) mediated
conditions or diseases are provided.
Inventors: |
CHEN; Xi; (East Palo Alto,
CA) ; DRAGOLI; Dean R.; (Los Altos, CA) ; FAN;
Pingchen; (Fremont, CA) ; LELETI; Manmohan Reddy;
(San Jose, CA) ; LUI; Rebecca M.; (Santa Clara,
CA) ; MALATHONG; Viengkham; (Mountain View, CA)
; POWERS; Jay P.; (Pacifica, CA) ; SINGH;
Rajinder; (Belmont, CA) ; TANAKA; Hiroko;
(Mountain View, CA) ; YANG; Ju; (Palo Alto,
CA) ; YU; Chao; (Sunnyvale, CA) ; ZHANG;
Penglie; (Foster City, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CHEMOCENTRYX, INC. |
Mountain View |
CA |
US |
|
|
Family ID: |
59314376 |
Appl. No.: |
15/408896 |
Filed: |
January 18, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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62280969 |
Jan 20, 2016 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 209/34 20130101;
C07D 401/04 20130101; C07D 405/04 20130101; A61P 35/00 20180101;
A61P 19/00 20180101; A61P 1/00 20180101; A61P 11/00 20180101; A61P
17/00 20180101; C07D 405/14 20130101 |
International
Class: |
C07D 405/14 20060101
C07D405/14; C07D 405/04 20060101 C07D405/04; C07D 401/04 20060101
C07D401/04; C07D 209/34 20060101 C07D209/34 |
Claims
1. A compound having formula (I): ##STR00136## or a
pharmaceutically acceptable salt thereof, wherein Ar is a 5- to
10-membered aromatic or heteroaromatic ring, optionally substituted
with from one to three R; L.sup.1 is selected from the group
consisting of a bond, C.sub.1-6 alkylene, and C.sub.1-6
heteroalkylene, L.sup.2 is selected from the group consisting of a
bond, C.sub.1-6 alkylene, and C.sub.1-6 heteroalkylene, Y is
CO.sub.2H or a carboxylic acid bioisostere; each R.sup.1 and each
R.sup.2a is independently selected from the group consisting of
halogen, cyano, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
haloalkyl, C.sub.1-6 haloalkoxy, C.sub.3-6 cycloalkyl, and
C.sub.2-6 alkenyl, wherein the alkyl, cycloalkyl and alkenyl
portions are optionally substituted with from one to three members
selected from fluoro, OH, CN, C.sub.1-3 alkyl, C.sub.1-3 haloalkyl,
and C.sub.1-3 alkoxy; R.sup.2b is selected from the group
consisting of H, halogen, cyano, C.sub.1-6 alkyl, C.sub.1-6 alkoxy,
C.sub.3-6 cycloalkyl, and C.sub.2-6 alkenyl, wherein the alkyl,
cycloalkyl and alkenyl portions are optionally substituted with
from one to three members selected from fluoro, CN, C.sub.1-3
alkyl, C.sub.1-3 haloalkyl, and C.sub.1-3 alkoxy; or optionally one
R.sup.2a and R.sup.2b when on adjacent vertices of a phenyl ring,
may be joined together to form a 5- or 6-membered heterocycloalkyl
ring having one or two ring vertices independently selected from O,
N and S, wherein said heterocycloalkyl ring is optionally
substituted with from one to three members selected from fluoro and
C.sub.1-3 alkyl; each R.sup.3 is independently selected from the
group consisting of halogen, cyano, C.sub.1-6 alkyl, C.sub.1-6
alkoxy, C.sub.1-6 haloalkyl, C.sub.1-6 haloalkoxy, C.sub.3-6
cycloalkyl, and C.sub.2-6 alkenyl; the subscript m is an integer of
from 0 to 4; and the subscript n is an integer of from 0 to 3.
2. The compound of claim 1, having formula (I'): ##STR00137## or a
pharmaceutically acceptable salt thereof, wherein Ar is a 5- to
10-membered aromatic or heteroaromatic ring, optionally substituted
with from one to three R.sup.3; L.sup.1 is selected from the group
consisting of a bond, C.sub.1-6 alkylene, and C.sub.1-6
heteroalkylene, L.sup.2 is selected from the group consisting of a
bond, C.sub.1-6 alkylene, and C.sub.1-6 heteroalkylene, each
R.sup.1 and each R.sup.2a is independently selected from the group
consisting of halogen, cyano, C.sub.1-6 alkyl, C.sub.1-6 alkoxy,
C.sub.1-6 haloalkyl, C.sub.1-6 haloalkoxy, C.sub.3-6 cycloalkyl,
and C.sub.2-6 alkenyl, wherein the alkyl, cycloalkyl and alkenyl
portions are optionally substituted with from one to three members
selected from fluoro, CN, C.sub.1-3 alkyl, C.sub.1-3 haloalkyl, and
C.sub.1-3 alkoxy; R.sup.2b is selected from the group consisting of
H, halogen, cyano, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.3-6
cycloalkyl, and C.sub.2-6 alkenyl, wherein the alkyl, cycloalkyl
and alkenyl portions are optionally substituted with from one to
three members selected from fluoro, CN, C.sub.1-3 alkyl, C.sub.1-3
haloalkyl, and C.sub.1-3 alkoxy; or optionally one R.sup.2a and
R.sup.2b when on adjacent vertices of a phenyl ring, may be joined
together to form a 5- or 6-membered heterocycloalkyl ring having
one or two ring vertices independently selected from O, N and S,
wherein said heterocycloalkyl ring is optionally substituted with
from one to three members selected from fluoro and C.sub.1-3 alkyl;
each R.sup.3 is independently selected from the group consisting of
halogen, cyano, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
haloalkyl, C.sub.1-6 haloalkoxy, C.sub.3-6 cycloalkyl, and
C.sub.2-6 alkenyl; the subscript m is an integer of from 0 to 4;
and the subscript n is an integer of from 0 to 3.
3. A compound of claim 1, wherein Y is selected from the group
consisting of: ##STR00138## ##STR00139## tetrazolyl and
tetrazolonyl, wherein the tetrazolyl or tetrazolonyl is optionally
substituted with R, wherein p is 0, 1 or 2 and wherein each R group
is independently selected from the group consisting of H,
C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6
hydroxyalkyl, C.sub.1-C.sub.6 alkoxy, or C.sub.1-C.sub.4
alkyl-O--C.sub.1-C.sub.4 alkyl.
4. A compound of claim 1, wherein Y is selected from the group
consisting of: tetrazolyl and tetrazolonyl, wherein the tetrazolyl
or tetrazolonyl is optionally substituted with C.sub.1-6 alkyl,
C.sub.1-6 haloalkyl, C.sub.1-6 hydroxyalkyl, --C.sub.1-6 alkoxy or
C.sub.1-4alkyl-O--C.sub.1-4alkyl.
5. A compound of claim 1, having the formula selected from the
group consisting of: ##STR00140## or a pharmaceutically acceptable
salt thereof, wherein said compound is substantially free of other
isomers.
6. A compound of claim 1, having the formula: ##STR00141## or a
pharmaceutically acceptable salt thereof, wherein L.sup.2 is
C.sub.1-3 alkylene and wherein said compound is substantially free
of other isomers.
7. A compound of claim 1, wherein Ar is selected from benzene,
pyridine and quinoline, each of which is optionally substituted
with from one to two R.sup.3.
8. A compound of claim 1, wherein L.sup.1 is selected from the
group consisting of a bond, --CH.sub.2-- and --CH(CH.sub.3)--.
9. A compound of claim 1, wherein L.sup.2 is selected from the
group consisting of a bond, --O--CH.sub.2--, --CH(CH.sub.3)--,
--C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2--, --CH.sub.2--, and
--CH.sub.2CH.sub.2CH.sub.2--.
10. A compound of claim 1, wherein n is 1 or 2.
11. A compound of claim 1, wherein m is 1, 2 or 3.
12. A compound of claim 1, having the formula: ##STR00142## or a
pharmaceutically acceptable salt thereof.
13. A compound of claim 12, wherein Ar is selected from benzene,
pyridine and quinoline, each of which is optionally substituted
with from one to two R.sup.3.
14. A compound of claim 12, wherein L.sup.1 is selected from the
group consisting of a bond, --CH.sub.2-- and --CH(CH.sub.3)--.
15. A compound of claim 12, wherein L.sup.2 is selected from the
group consisting of a bond, --O--CH.sub.2--, --CH(CH.sub.3)--,
--C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2--, --CH.sub.2--, and
--CH.sub.2CH.sub.2CH.sub.2--.
16. A compound of claim 12, having a formula selected from the
group consisting of: ##STR00143## or a pharmaceutically acceptable
salt thereof, wherein said compound is substantially free of other
isomers.
17. A compound of claim 16, wherein Ar is selected from the group
consisting of benzene, pyridine and quinoline, each of which is
optionally substituted with from one to two R.sup.3.
18. A compound of claim 16, wherein Ar is selected from the group
consisting of 1,3-phenylene and 1,4-phenylene, each of which is
optionally substituted with from one to two R.sup.3.
19. A compound of claim 16, wherein L.sup.1 is selected from the
group consisting of a bond, --CH.sub.2-- and --CH(CH.sub.3)--.
20. A compound of claim 16, wherein L.sup.2 is selected from the
group consisting of --O--CH.sub.2--, --CH(CH.sub.3)--,
--C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2--, --CH.sub.2-- and
--CH.sub.2CH.sub.2CH.sub.2--.
21. A compound of claim 16, wherein R.sup.1 is selected from the
group consisting of halogen, cyano, C.sub.1-3 alkyl, C.sub.1-3
alkoxy, C.sub.1-3 haloalkyl, C.sub.1-3 haloalkoxy, C.sub.3-5
cycloalkyl, and C.sub.2-3 alkenyl.
22. A compound of claim 16, wherein R.sup.1 is selected from the
group consisting of chloro, methyl, cyano, ethyl, cyclopropyl, 3
trifluoromethyl and trifluoromethoxy.
23. A compound of claim 16, having a formula selected from the
group consisting of: ##STR00144## or a pharmaceutically acceptable
salt thereof, wherein said compound is substantially free of other
isomers.
24. A compound of claim 1, having the formula: ##STR00145## or a
pharmaceutically acceptable salt thereof.
25. A compound of claim 24, wherein R.sup.2b is hydrogen.
26. A compound of claim 24, wherein Ar is selected from the group
consisting of benzene, pyridine and quinoline, each of which is
optionally substituted with from one to two R.sup.3.
27. A compound of claim 24, wherein L.sup.1 is selected from the
group consisting of a bond, --CH.sub.2-- and --CH(CH.sub.3)--.
28. A compound of claim 24, wherein L.sup.2 is selected from the
group consisting of a bond, --O--CH.sub.2--, --CH(CH.sub.3)--,
--C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2--, --CH.sub.2-- and
--CH.sub.2CH.sub.2CH.sub.2--.
29. A compound of claim 24, having a formula selected from the
group consisting of: ##STR00146## or a pharmaceutically acceptable
salt thereof, wherein said compound is substantially free of other
isomers.
30. A compound of claim 29, wherein Ar is selected from the group
consisting of benzene, pyridine and quinoline, each of which is
optionally substituted with from one to two R.sup.3.
31. A compound of claim 29, wherein Ar is selected from the group
consisting of 1,3-phenylene and 1,4-phenylene, each of which is
optionally substituted with from one to two R.sup.3.
32. A compound of claim 29, wherein R.sup.3 is selected from the
group consisting of CH.sub.3, CH.sub.2CH.sub.3,
CH.sub.2CH.sub.2CH.sub.3, CH(CH.sub.3).sub.2 and CH.sub.2OH.
33. A compound claim 29, wherein L.sup.1 is selected from the group
consisting of a bond, --CH.sub.2-- and --CH(CH.sub.3)--.
34. A compound of claim 29, wherein L.sup.2 is selected from the
group consisting of --O--CH.sub.2--, --CH(CH.sub.3)--,
--C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2--, --CH.sub.2-- and
--CH.sub.2CH.sub.2CH.sub.2--.
35. A compound of claim 29, wherein R.sup.1 is selected from the
group consisting of halogen, cyano, C.sub.1-3 alkyl, C.sub.1-3
alkoxy, C.sub.1-3 haloalkyl, C.sub.1-3 haloalkoxy, C.sub.3-5
cycloalkyl, and C.sub.2-3 alkenyl.
36. A compound of claim 29, wherein R.sup.1 is selected from the
group consisting of chloro, methyl, cyano, ethyl, cyclopropyl,
trifluoromethyl and trifluoromethoxy.
37. A compound of claim 29, having a formula selected from the
group consisting of: ##STR00147## or a pharmaceutically acceptable
salt thereof, wherein said compound is substantially free of other
isomers.
38. A compound of claim 1 or a pharmaceutically acceptable salt
thereof, selected from Table 1 and having activity of ++ or
+++.
39. A compound of claim 1 or a pharmaceutically acceptable salt
thereof, selected from Table 1 and having activity of +++.
40. A pharmaceutical composition comprising a pharmaceutically
acceptable excipient and at least one compound of claim 1, or a
pharmaceutically acceptable salt thereof.
41. A method of treating a CCR(9) mediated disease or condition in
a subject in need thereof, said method comprising administering to
said subject an effective amount of a compound of claim 1 or a
pharmaceutically acceptable salt thereof, optionally in admixture
with a pharmaceutically acceptable excipient or carrier.
42. A method in accordance with claim 41, wherein said disease or
condition is selected from the group consisting of inflammatory
bowel diseases, an allergic disease, psoriasis, atopic dermatitis,
asthma, fibrotic diseases, graft rejection, immune mediated food
allergies, autoimmune diseases, Celiac disease, rheumatoid
arthritis, thymoma, thymic carcinoma, leukemia, Sjogren syndrome,
GvHD (graft versus host disease), graft rejection, solid tumor, or
acute lymphocytic leukemia, melanoma, primary sclerosing
cholangitis, hepatitis and inflammatory hepatic disease or
post-operative ileus.
43. A method in accordance with claim 41, wherein said disease or
condition is selected from the group consisting of inflammatory
bowel diseases.
44. A method in accordance with claim 41, wherein said disease or
condition is selected from the group consisting of Crohn's disease
and ulcerative colitis.
Description
CROSS-REFERENCES TO RELATED APPLICATIONS
[0001] This application is an application claiming benefit under 35
U.S.C. .sctn.119(e) of U.S. Provisional Application No. 62/280,969
filed Jan. 20, 2016, which application is herein incorporated by
reference in its entirety for all purposes.
STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED
RESEARCH AND DEVELOPMENT
[0002] NOT APPLICABLE
REFERENCE TO A "SEQUENCE LISTING," A TABLE, OR A COMPUTER PROGRAM
LISTING APPENDIX SUBMITTED ON A COMPACT DISK
[0003] NOT APPLICABLE
BACKGROUND OF THE INVENTION
[0004] The present disclosure provides compounds and pharmaceutical
compositions containing one or more of those compounds or their
pharmaceutically acceptable salts, that are effective in inhibiting
the binding or function of chemokines to the CCR9 chemokine
receptor. As antagonists or modulators of the CCR9 chemokine
receptor, the compounds and compositions can have utility in
treating various immune disorder conditions and diseases.
[0005] Chemokines, also known as chemotactic cytokines, are a group
of small molecular-weight proteins that are released by a wide
variety of cells and have a variety of biological activities.
Chemokines attract various types of cells of the immune system,
such as macrophages, T cells, eosinophils, basophils and
neutrophils, and cause them to migrate from the blood to various
lymphoid and none-lymphoid tissues. They mediate infiltration of
inflammatory cells to sites of inflammation, and are responsible
for the initiation and perpetuation of many inflammation diseases
(reviewed in Schall, Cytokine, 3:165-183 (1991), Schall et al.,
Curr. Opin. Immunol., 6:865-873 (1994)).
[0006] In addition to stimulating chemotaxis, chemokines can induce
other changes in responsive cells, including changes in cell shape,
granule exocytosis, integrin up-regulation, formation of bioactive
lipids (e.g., leukotrienes), respiratory burst associated with
leukocyte activation, cell proliferation, resistance to induction
of apoptosis and angiogenesis. Thus, chemokines are early triggers
of the inflammatory response, causing inflammatory mediator
release, chemotaxis and extravasation to sites of infection or
inflammation. They are also stimulators of a multitude of cellular
processes that bear important physiological functions as well as
pathological consequences.
[0007] Chemokines exert their effects by activating chemokine
receptors expressed by responsive cells. Chemokine receptors are a
class of G-protein coupled receptors, also known as
seven-transmembrane receptors, found on the surface of a wide
variety of cell types such as leukocytes, endothelial cells, smooth
muscle cells and tumor cells.
[0008] Chemokines and chemokine receptors are expressed by
intrinsic renal cells and infiltrating cells during renal
inflammation (Segerer et al., J. Am. Soc. Nephrol., 11:152-76
(2000); Morii et al., J. Diabetes Complications, 17:11-5 (2003);
Lloyd et al. J. Exp. Med., 185:1371-80 (1997); Gonzalez-Cuadrado et
al. Clin. Exp. Immunol., 106:518-22 (1996); Eddy & Giachelli,
Kidney Int., 47:1546-57 (1995); Diamond et al., Am. J. Physiol.,
266:F926-33 (1994)).
[0009] T lymphocyte (T cell) infiltration into the small intestine
and colon has been linked to the pathogenesis of Coeliac diseases,
food allergies, rheumatoid arthritis, human inflammatory bowel
diseases (IBD) which include Crohn's disease and ulcerative
colitis. Blocking trafficking of relevant T cell populations to the
intestine can lead to an effective approach to treat human IBD.
More recently, chemokine receptor-9 (CCR(9)) has been noted to be
expressed on gut-homing T cells in peripheral blood, elevated in
patients with small bowel inflammation such as Crohn's disease and
coeliac disease. The only CCR(9) ligand identified to date, TECK
(thymus-expressed chemokine) is expressed in both the small and
large intestines and the ligand receptor pair is now thought to
play a pivotal role in the development of IBD. In particular, this
pair mediates the migration of disease causing inflammatory cells
to the intestine. See for example, Zaballos et al., J. Immunol.,
162(10):5671-5675 (1999); Kunkel et al., J. Exp. Med.,
192(5):761-768 (2000); Papadakis et al., J. Immunol.,
165(9):5069-5076 (2000); Papadakis et al., Gastroenterology,
121(2):246-254 (2001); Campbell et al., J. Exp. Med.,
195(1):135-141 (2002); Wurbel et al., Blood, 98(9):2626-2632
(2001); and Uehara et al., J. Immunol, 168(6):2811-2819 (2002);
Rivera-Nieves et al., Gastroenterology, 2006 November; 131(5):
1518-29; and Kontoyiannis et al., J. Exp. Med., Vol. 196, Number
12, Dec. 16, 2002. In addition CCR(9) bearing lymphocytes have been
show to mediate the pathology of filariasis (lymphatic filarial
disease) and inhibition of CCR(9) has been correlated with
reduction of the pathology associated with such conditions. See for
example Babu et al., Journal of Infectious Diseases, 191: 1018-26,
2005.
[0010] The identification of compounds that modulate the function
of CCR(9) represents an attractive new family of therapeutic agents
for the treatment of inflammatory and other conditions and diseases
associated with CCR(9) activation, such as inflammatory bowel
disease.
BRIEF SUMMARY OF THE INVENTION
[0011] The present invention is directed to compounds and
pharmaceutically acceptable salts thereof, compositions, and
methods useful in modulating the function of CCR(9). The compounds
and salts thereof, compositions, and methods described herein can
be useful in treating or preventing chemokine-mediated conditions
or diseases, including certain inflammatory and immunoregulatory
disorders and diseases.
[0012] The compounds of the present disclosure have been shown to
modulate CCR(9), as shown in the examples.
[0013] In one aspect, the present compounds may be represented by
formula (I):
##STR00001##
or a pharmaceutically acceptable salt thereof, wherein [0014] Ar is
a 5- to 10-membered aromatic or heteroaromatic ring, optionally
substituted with from one to three R.sup.3; [0015] L.sup.1 is
selected from the group consisting of a bond, C.sub.1-6 alkylene,
and C.sub.1-6 heteroalkylene, [0016] L.sup.2 is selected from the
group consisting of a bond, C.sub.1-6 alkylene, and C.sub.1-6
heteroalkylene, [0017] Y is CO.sub.2H or a carboxylic acid
bioisostere; [0018] each R.sup.1 and each R.sup.2a is independently
selected from the group consisting of halogen, cyano, C.sub.1-6
alkyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, C.sub.1-6 haloalkoxy,
C.sub.3-6 cycloalkyl, and C.sub.2-6 alkenyl, wherein the alkyl,
cycloalkyl and alkenyl portions are optionally substituted with
from one to three members selected from fluoro, OH, CN, C.sub.1-3
alkyl, C.sub.1-3 haloalkyl, and C.sub.1-3 alkoxy; [0019] R.sup.2b
is selected from the group consisting of H, halogen, cyano,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.3-6 cycloalkyl, and
C.sub.2-6 alkenyl, wherein the alkyl, cycloalkyl and alkenyl
portions are optionally substituted with from one to three members
selected from fluoro, CN, C.sub.1-3 alkyl, C.sub.1-3 haloalkyl, and
C.sub.1-3 alkoxy; [0020] or optionally one R.sup.2a and R.sup.2b
when on adjacent vertices of a phenyl ring, may be joined together
to form a 5- or 6-membered heterocycloalkyl ring having one or two
ring vertices independently selected from O, N and S, wherein said
heterocycloalkyl ring is optionally substituted with from one to
three members selected from fluoro and C.sub.1-3 alkyl; [0021] each
R.sup.3 is independently selected from the group consisting of
halogen, cyano, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
haloalkyl, C.sub.1-6 haloalkoxy, C.sub.3-6 cycloalkyl, and
C.sub.2-6 alkenyl; [0022] the subscript m is an integer of from 0
to 4; and [0023] the subscript n is an integer of from 0 to 3.
[0024] In another aspect, the present disclosure provides
compositions useful in modulating chemokine activity. In one
embodiment, a composition according to the present disclosure
comprises a compound according to the disclosure and a
pharmaceutically acceptable carrier or excipient.
[0025] In yet another aspect, the present disclosure provides
methods of modulating chemokine function in a cell, comprising
contacting the cell with a therapeutically effective amount of a
compound or composition according to the disclosure.
[0026] In still another aspect, the present disclosure provides
methods for modulating chemokine function, comprising contacting a
chemokine receptor with a therapeutically effective amount of a
compound or composition according to the disclosure.
[0027] In still another aspect, the present disclosure provides
methods for treating a chemokine-mediated condition or disease,
comprising administering to a subject a safe and effective amount
of a compound or composition according to the disclosure. The
administering may be oral, parenteral, rectal, transdermal,
sublingual, nasal or topical. In some aspects the compound may be
administered in combination with an anti-inflammatory or analgesic
agent.
[0028] In addition to the compounds provided herein, the present
disclosure further provides pharmaceutical compositions containing
one or more of these compounds, as well as methods for the use of
these compounds in therapeutic methods, primarily to treat diseases
associated with chemokine signaling activity. The CCR(9) mediated
disease or condition may be inflammatory bowel diseases, an
allergic disease, psoriasis, atopic dermatitis, asthma, fibrotic
diseases, graft rejection, GvHD, Sjogren syndrome, immune mediated
food allergies, autoimmune diseases, Celiac disease, rheumatoid
arthritis, thymoma, thymic carcinoma, leukemia, solid tumor, or
acute lymphocytic leukemia, melanoma, primary sclerosing
cholangitis, hepatitis and inflammatory hepatic disease,
post-operative ileus, Crohn's disease or ulcerative colitis.
BRIEF DESCRIPTION OF THE DRAWINGS
[0029] NOT APPLICABLE
DETAILED DESCRIPTION OF THE INVENTION
General
[0030] The present disclosure is directed to compounds and salts
thereof, compositions and methods useful in the modulation of
chemokine receptor function, particularly CCR(9) function.
Modulation of chemokine receptor activity, as used herein in its
various forms, is intended to encompass antagonism, agonism,
partial antagonism, inverse agonism and/or partial agonism of the
activity associated with a particular chemokine receptor,
preferably the CCR(9) receptor. Accordingly, the compounds of the
present disclosure are compounds which modulate at least one
function or characteristic of mammalian CCR(9), for example, a
human CCR(9) protein. The ability of a compound to modulate the
function of CCR(9), can be demonstrated in a binding assay (e.g.,
ligand binding or agonist binding), a chemotaxis (migration assay),
a signaling assay (e.g., activation of a mammalian G protein,
induction of rapid and transient increase in the concentration of
cytosolic free calcium), and/or cellular response assay (e.g.,
stimulation of chemotaxis, exocytosis or inflammatory mediator
release by leukocytes).
Abbreviations and Definitions
[0031] The term "alkyl", by itself or as part of another
substituent, means, unless otherwise stated, a straight or branched
chain hydrocarbon radical, having the number of carbon atoms
designated (i.e. C.sub.1-8 means one to eight carbons). Examples of
alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl,
t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,
and the like. The term "alkenyl" refers to an unsaturated alkyl
group having one or more double bonds. Similarly, the term
"alkynyl" refers to an unsaturated alkyl group having one or more
triple bonds. Examples of such unsaturated alkyl groups include
vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),
2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,
3-butynyl, and the higher homologs and isomers. The term
"cycloalkyl" refers to hydrocarbon rings having the indicated
number of ring atoms (e.g., C.sub.3-6cycloalkyl) and being fully
saturated or having no more than one double bond between ring
vertices. "Cycloalkyl" is also meant to refer to bicyclic and
polycyclic hydrocarbon rings such as, for example,
bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term
"heterocycloalkane" or "heterocycloalkyl" refers to a cycloalkyl
group that contain from one to five heteroatoms selected from N, O,
and S, wherein the nitrogen and sulfur atoms are optionally
oxidized, and the nitrogen atom(s) are optionally quaternized. The
heterocycloalkane may be a monocyclic, a bicyclic or a polycylic
ring system. Non limiting examples of heterocycloalkane groups
include pyrrolidine, imidazolidine, pyrazolidine, butyrolactam,
valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide,
piperidine, 1,4-dioxane, morpholine, thiomorpholine,
thiomorpholine-S-oxide, thiomorpholine-S,S-oxide, piperazine,
pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran,
tetrhydrothiophene, quinuclidine, and the like. A heterocycloalkane
group can be attached to the remainder of the molecule through a
ring carbon or a heteroatom.
[0032] The term "alkylene" by itself or as part of another
substituent means a divalent radical derived from an alkane, as
exemplified by --CH.sub.2CH.sub.2CH.sub.2CH.sub.2--. Typically, an
alkyl (or alkylene) group will have from 1 to 24 carbon atoms, with
those groups having 10 or fewer carbon atoms being preferred in the
present disclosure. A "lower alkyl" or "lower alkylene" is a
shorter chain alkyl or alkylene group, generally having four or
fewer carbon atoms. Similarly, "alkenylene" and "alkynylene" refer
to the unsaturated forms of "alkylene" having double or triple
bonds, respectively. The term "heteroalkylene" refers to an
alkylene group in which one or two carbon atoms are replaced by N,
O, or S.
[0033] As used herein, a wavy line, "", that intersects a single,
double or triple bond in any chemical structure depicted herein,
represent the point attachment of the single, double, or triple
bond to the remainder of the molecule.
[0034] The terms "alkoxy," "alkylamino" and "alkylthio" (or
thioalkoxy) are used in their conventional sense, and refer to
those alkyl groups attached to the remainder of the molecule via an
oxygen atom, an amino group, or a sulfur atom, respectively.
Additionally, for dialkylamino groups, the alkyl portions can be
the same or different and can also be combined to form a 3-7
membered ring with the nitrogen atom to which each is attached.
Accordingly, a group represented as dialkylamino or
--NR.sup.aR.sup.b is meant to include piperidinyl, pyrrolidinyl,
morpholinyl, azetidinyl and the like.
[0035] The term "di-(C.sub.1-4 alkyl)amino-C.sub.1-4 alkyl" refers
to an amino group bearing two C.sub.1-4 alkyl groups that can be
the same or different (e.g., methyl, ethyl, propyl, isopropyl,
n-butyl, sec-butyl, isobutyl and tert-butyl) and which is attached
to the remainder of the molecule through a C.sub.1-4 alkyl group (a
one to four carbon alkylene linking group). Examples of
di-(C.sub.1-4 alkyl)amino-C.sub.1-4 alkyl groups include
dimethylaminomethyl, 2-(ethyl(methyl)amino)ethyl,
3-(dimethylamino)butyl, and the like.
[0036] The terms "halo" or "halogen," by themselves or as part of
another substituent, mean, unless otherwise stated, a fluorine,
chlorine, bromine, or iodine atom. Additionally, terms such as
"haloalkyl" and "haloalkoxy," are meant to include monohalo- and
polyhalo-versions of alkyl and alkoxy, respectively. For example,
the term "C.sub.1-4 haloalkyl" is mean to include trifluoromethyl,
2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the
like.
[0037] The term "aryl" or "aromatic ring" means, unless otherwise
stated, a polyunsaturated, typically aromatic, hydrocarbon group
which can be a single ring or multiple rings (up to three rings)
which are fused together or linked covalently. Similarly, the terms
"heteroaryl" and "heteroaromatic ring" refer to aryl groups (or
rings) that contain from one to five heteroatoms selected from N,
O, and S, wherein the nitrogen and sulfur atoms are optionally
oxidized, and the nitrogen atom(s) are optionally quaternized. A
heteroaryl group or heteroaromatic ring can be attached to the
remainder of the molecule through a heteroatom. Non-limiting
examples of aryl groups include phenyl, naphthyl and biphenyl,
while non-limiting examples of heteroaryl groups include pyridyl,
pyridazinyl, pyrazinyl, pyrimindinyl, triazinyl, quinolinyl,
quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl,
benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl,
benzotriazolyl, benzisoxazolyl, isobenzofuryl, isoindolyl,
indolizinyl, benzotriazinyl, thienopyridinyl, thienopyrimidinyl,
pyrazolopyrimidinyl, imidazopyridines, benzothiaxolyl,
benzofuranyl, benzothienyl, indolyl, quinolyl, isoquinolyl,
isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl,
triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl,
pyrrolyl, thiazolyl, furyl, thienyl and the like. Substituents for
each of the above noted aryl and heteroaryl ring systems are
selected from the group of acceptable substituents described
below.
[0038] The term "arylalkyl" is meant to include those radicals in
which an aryl group is attached to an alkyl group (e.g., benzyl,
phenethyl, and the like). Similarly, the term "heteroaryl-alkyl" is
meant to include those radicals in which a heteroaryl group is
attached to an alkyl group (e.g., pyridylmethyl, thiazolylethyl,
and the like).
[0039] As used herein, the term "heteroatom" is meant to include
oxygen (O), nitrogen (N), sulfur (S) and silicon (Si).
[0040] The term "carboxylic acid bioisostere" refers to a group
having polar and/or acidic character to act as a replacement for a
carboxylic acid. A variety of bioisosteres are known for carboxylic
acids, including, hydroxamic acids, hydroxamic esters, phosphonic
acids, phosphinic acids, sulfonic acids, sulfinic acids,
sulfonamides, acyl sulfonamides, acylureas, sulfonylureas,
cyclopentane-1,2-diones, substituted phenols, and heterocycle-based
bioisosteres as provided below:
##STR00002## ##STR00003##
wherein p is 0, 1 or 2 and wherein each R group is independently
selected from the group consisting of H, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 hydroxyalkyl,
C.sub.1-C.sub.6 alkoxy, or C.sub.1-C.sub.4 alkyl-O--C.sub.1-C.sub.4
alkyl.
[0041] Other examples of bioisosteres of a carboxylic acid are a
tetrazolyl or tetrazolonyl, wherein the tetrazolyl or tetrazolonyl
is optionally substituted with C.sub.1-6 alkyl, C.sub.1-6
haloalkyl, C.sub.1-6 hydroxyalkyl, --C.sub.1-6 alkoxy or
C.sub.1-4alkyl-O--C.sub.1-4alkyl. Still other examples of
carboxylic acid bioisosteres are described in the Journal of
Medicinal Chemistry, 2016, 59, 3183-3203 which are hereby
incorporated by reference.
[0042] The term "pharmaceutically acceptable salts" is meant to
include salts of the active compounds which are prepared with
relatively nontoxic acids or bases, depending on the particular
substituents found on the compounds described herein. When
compounds of the present disclosure contain relatively acidic
functionalities, base addition salts can be obtained by contacting
the neutral form of such compounds with a sufficient amount of the
desired base, either neat or in a suitable inert solvent. Examples
of salts derived from pharmaceutically-acceptable inorganic bases
include aluminum, ammonium, calcium, copper, ferric, ferrous,
lithium, magnesium, manganic, manganous, potassium, sodium, zinc
and the like. Salts derived from pharmaceutically-acceptable
organic bases include salts of primary, secondary and tertiary
amines, including substituted amines, cyclic amines,
naturally-occurring amines and the like, such as arginine, betaine,
caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine,
2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,
ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,
glucosamine, histidine, hydrabamine, isopropylamine, lysine,
methylglucamine, morpholine, piperazine, piperidine, polyamine
resins, procaine, purines, theobromine, triethylamine,
trimethylamine, tripropylamine, tromethamine and the like. When
compounds of the present disclosure contain relatively basic
functionalities, acid addition salts can be obtained by contacting
the neutral form of such compounds with a sufficient amount of the
desired acid, either neat or in a suitable inert solvent. Examples
of pharmaceutically acceptable acid addition salts include those
derived from inorganic acids like hydrochloric, hydrobromic,
nitric, carbonic, monohydrogencarbonic, phosphoric,
monohydrogenphosphoric, dihydrogenphosphoic, sulfuric,
monohydrogensulfuric, hydriodic, or phosphorous acids and the like,
as well as the salts derived from relatively nontoxic organic acids
like acetic, propionic, isobutyric, malonic, benzoic, succinic,
suberic, fumaric, mandelic, phthalic, benzenesulfonic,
p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like.
Also included are salts of amino acids such as arginate and the
like, and salts of organic acids like glucuronic or galactunoric
acids and the like (see, for example, Berge, S. M., et al,
"Pharmaceutical Salts", Journal of Pharmaceutical Science, 1977,
66, 1-19). Certain specific compounds of the present disclosure
contain both basic and acidic functionalities that allow the
compounds to be converted into either base or acid addition
salts.
[0043] The neutral forms of the compounds may be regenerated by
contacting the salt with a base or acid and isolating the parent
compound in the conventional manner. The parent form of the
compound differs from the various salt forms in certain physical
properties, such as solubility in polar solvents, but otherwise the
salts are equivalent to the parent form of the compound for the
purposes of the present disclosure.
[0044] In addition to salt forms, the present disclosure provides
compounds which are in a prodrug form. Prodrugs of the compounds
described herein are those compounds that readily undergo chemical
changes under physiological conditions to provide the compounds of
the present disclosure. Additionally, prodrugs can be converted to
the compounds of the present disclosure by chemical or biochemical
methods in an ex vivo environment. For example, prodrugs can be
slowly converted to the compounds of the present disclosure when
placed in a transdermal patch reservoir with a suitable enzyme or
chemical reagent.
[0045] Certain compounds of the present disclosure can exist in
unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms are equivalent to unsolvated
forms and are intended to be encompassed within the scope of the
present disclosure. Certain compounds of the present disclosure may
exist in multiple crystalline or amorphous forms. In general, all
physical forms are equivalent for the uses contemplated by the
present disclosure and are intended to be within the scope of the
present disclosure.
[0046] Certain compounds of the present disclosure possess
asymmetric carbon atoms (optical centers) or double bonds; the
racemates, diastereomers, geometric isomers, regioisomers and
individual isomers (e.g., separate enantiomers) are all intended to
be encompassed within the scope of the present disclosure. When a
stereochemical depiction is shown, it is meant to refer the
compound in which one of the isomers is present and substantially
free of the other isomer. `Substantially free of` another isomer
indicates at least an 80/20 ratio of the two isomers, more
preferably 90/10, or 95/5 or more. In some embodiments, one of the
isomers will be present in an amount of at least 99%.
[0047] The compounds of the present disclosure may also contain
unnatural proportions of atomic isotopes at one or more of the
atoms that constitute such compounds. Unnatural proportions of an
isotope may be defined as ranging from the amount found in nature
to an amount consisting of 100% of the atom in question. For
example, the compounds may incorporate radioactive isotopes, such
as for example tritium (.sup.3H), iodine-125 (.sup.125I) or
carbon-14 (.sup.14C), or non-radioactive isotopes, such as
deuterium (.sup.2H) or carbon-13 (.sup.13C). Such isotopic
variations can provide additional utilities to those described
elsewhere with this application. For instance, isotopic variants of
the compounds of the disclosure may find additional utility,
including but not limited to, as diagnostic and/or imaging
reagents, or as cytotoxic/radiotoxic therapeutic agents.
Additionally, isotopic variants of the compounds of the disclosure
can have altered pharmacokinetic and pharmacodynamic
characteristics which can contribute to enhanced safety,
tolerability or efficacy during treatment. All isotopic variations
of the compounds of the present disclosure, whether radioactive or
not, are intended to be encompassed within the scope of the present
disclosure. Substitution with heavier isotopes such as deuterium,
i.e. .sup.2H, may afford certain therapeutic advantages resulting
from greater metabolic stability. For example, in vivo half-life
may increase or dosage requirements may be reduced.
[0048] Compounds of the disclosure having formula I can exist in
different isomeric forms. As used herein, the terms cis or trans
are used in their conventional sense in the chemical arts, i.e.,
referring to the position of the substituents to one another
relative to a reference plane, e.g., a double bond, or a ring
system, such as a decalin-type ring system or a hydroquinolone ring
system: in the cis isomer, the substituents are on the same side of
the reference plane, in the trans isomer the substituents are on
opposite sides. Additionally, different conformers are contemplated
by the present disclosure, as well as distinct rotamers. Conformers
are conformational isomers that can differ by rotations about one
or more a bonds. Rotamers are conformers that differ by rotation
about only a single a bond.
Compounds
[0049] The present disclosure provides compounds that modulate the
activity of CCR(9). Chemokine receptors are integral membrane
proteins which interact with an extracellular ligand, such as a
chemokine, and mediate a cellular response to the ligand, e.g.,
chemotaxis, increased intracellular calcium ion concentration, etc.
Therefore, modulation of a chemokine receptor function, e.g.,
interference with a chemokine receptor ligand interaction, will
modulate a chemokine receptor mediated response, and treat or
prevent a chemokine receptor mediated condition or disease.
Modulation of a chemokine receptor function includes both
inducement and inhibition of the function. The type of modulation
accomplished will depend on the characteristics of the compound,
i.e., antagonist or full, partial or inverse agonist.
[0050] For example, compounds of this disclosure act as potent
CCR(9) antagonists, and this antagonistic activity has been further
confirmed in animal testing for inflammation, one of the hallmark
disease states for CCR(9). Accordingly, the compounds provided
herein are useful in pharmaceutical compositions, methods for the
treatment of CCR(9)-mediated diseases, and as controls in assays
for the identification of competitive CCR(9) antagonists.
[0051] Provided herein are compounds of formula (I):
##STR00004##
or a pharmaceutically acceptable salt thereof, wherein [0052] Ar is
a 5- to 10-membered aromatic or heteroaromatic ring, optionally
substituted with from one to three R.sup.1; [0053] L.sup.1 is
selected from the group consisting of a bond, C.sub.1-6 alkylene,
and C.sub.1-6 heteroalkylene, [0054] L.sup.2 is selected from the
group consisting of a bond, C.sub.1-6 alkylene, and C.sub.1-6
heteroalkylene, [0055] Y is CO.sub.2H or a carboxylic acid
bioisostere; [0056] each R.sup.1 and each R.sup.2a is independently
selected from the group consisting of halogen, cyano, C.sub.1-6
alkyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, C.sub.1-6haloalkoxy,
C.sub.3-6 cycloalkyl, and C.sub.2-6 alkenyl, wherein the alkyl,
cycloalkyl and alkenyl portions are optionally substituted with
from one to three members selected from fluoro, OH, CN, C.sub.1-3
alkyl, C.sub.1-3 haloalkyl, and C.sub.1-3 alkoxy; [0057] R.sup.2b
is selected from the group consisting of H, halogen, cyano,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.3-6 cycloalkyl, and
C.sub.2-6 alkenyl, wherein the alkyl, cycloalkyl and alkenyl
portions are optionally substituted with from one to three members
selected from fluoro, CN, C.sub.1-3 alkyl, C.sub.1-3 haloalkyl, and
C.sub.1-3 alkoxy; [0058] or optionally one R.sup.2a and R.sup.2b
when on adjacent vertices of a phenyl ring, may be joined together
to form a 5- or 6-membered heterocycloalkyl ring having one or two
ring vertices independently selected from O, N and S, wherein said
heterocycloalkyl ring is optionally substituted with from one to
three members selected from fluoro and C.sub.1-3 alkyl; [0059] each
R.sup.3 is independently selected from the group consisting of
halogen, cyano, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
haloalkyl, C.sub.1-6 haloalkoxy, C.sub.3-6 cycloalkyl, and
C.sub.2-6 alkenyl; [0060] the subscript m is an integer of from 0
to 4; and [0061] the subscript n is an integer of from 0 to 3.
[0062] In some embodiments, Y is selected from the group consisting
of:
##STR00005## ##STR00006##
tetrazolyl and tetrazolonyl, wherein the tetrazolyl or tetrazolonyl
is optionally substituted with R, wherein p is 0, 1 or 2 and
wherein each R group is independently selected from the group
consisting of H, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl,
C.sub.1-C.sub.6 hydroxyalkyl, C.sub.1-C.sub.6 alkoxy, or
C.sub.1-C.sub.4 alkyl-O--C.sub.1-C.sub.4 alkyl.
[0063] In some embodiments, Y is selected from the group consisting
of: tetrazolyl and tetrazolonyl, wherein the tetrazolyl or
tetrazolonyl is optionally substituted with C.sub.1-6 alkyl,
C.sub.1-6 haloalkyl, C.sub.1-6 hydroxyalkyl, --C.sub.1-6 alkoxy or
C.sub.1-4alkyl-O--C.sub.1-4alkyl.
[0064] In some embodiments, a compound is provided having the
formula selected from the group consisting of:
##STR00007##
or a pharmaceutically acceptable salt thereof, wherein said
compound is substantially free of other isomers.
[0065] In some embodiments, a compound is provided having the
formula:
##STR00008##
or a pharmaceutically acceptable salt thereof, wherein L.sup.2 is
C.sub.1-3 alkylene and wherein said compound is substantially free
of other isomers.
[0066] In some embodiments, a compound is provided having the
formula (I'):
##STR00009##
or a pharmaceutically acceptable salt thereof, wherein [0067] Ar is
a 5- to 10-membered aromatic or heteroaromatic ring, optionally
substituted with from one to three R.sup.3; [0068] L.sup.1 is
selected from the group consisting of a bond, C.sub.1-6 alkylene,
and C.sub.1-6 heteroalkylene, [0069] L.sup.2 is selected from the
group consisting of a bond, C.sub.1-6 alkylene, and C.sub.1-6
heteroalkylene, [0070] each R.sup.1 and each R.sup.2a is
independently selected from the group consisting of halogen, cyano,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, C.sub.1-6
haloalkoxy, C.sub.3-6 cycloalkyl, and C.sub.2-6 alkenyl, wherein
the alkyl, cycloalkyl and alkenyl portion are optionally
substituted with from one to three members selected from fluoro,
CN, C.sub.1-3 alkyl, C.sub.1-3 haloalkyl, and C.sub.1-3 alkoxy;
[0071] R.sup.2b is selected from the group consisting of H,
halogen, cyano, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.3-6
cycloalkyl, and C.sub.2-6 alkenyl, wherein the alkyl, cycloalkyl
and alkenyl portion are optionally substituted with from one to
three members selected from fluoro, CN, C.sub.1-3 alkyl, C.sub.1-3
haloalkyl, and C.sub.1-3 alkoxy; [0072] or optionally one R.sup.2a
and R.sup.2b when on adjacent vertices of a phenyl ring, may be
joined together to form a 5- or 6-membered cycloheteroalkyl ring
having one or two ring vertices independently selected from O, N
and S, wherein said cycloheteroalkyl ring is optionally substituted
with from one to three members selected from fluoro and C.sub.1-3
alkyl; [0073] each R.sup.3 is independently selected from the group
consisting of halogen, cyano, C.sub.1-6 alkyl, C.sub.1-6 alkoxy,
C.sub.1-6 haloalkyl, C.sub.1-6 haloalkoxy, C.sub.3-6 cycloalkyl,
and C.sub.2-6 alkenyl; [0074] the subscript m is an integer of from
0 to 4; and [0075] the subscript n is an integer of from 0 to
3.
[0076] In one group of embodiments for each of formulae (I) and
(I'), Ar is selected from benzene, pyridine and quinoline, each of
which is optionally substituted with from one to two R.sup.3.
[0077] In some selected embodiments of formula (I) and (I'),
L.sup.1 is selected from the group consisting of a bond,
--CH.sub.2-- and --CH(CH.sub.3)--. In other selected embodiments of
formula (I), L.sup.2 is selected from the group consisting of a
bond, --O--CH.sub.2--, --CH(CH.sub.3)--, --C(CH.sub.3).sub.2--,
--CH.sub.2CH.sub.2--, --CH.sub.2--, and
--CH.sub.2CH.sub.2CH.sub.2--.
[0078] In some selected embodiments of formula (I) and (I'), n is 1
or 2. In other selected embodiments of formula (I) and (I'), m is
1, 2 or 3.
[0079] In other embodiments, suitable compounds are provided having
the formula (Ia):
##STR00010##
or a pharmaceutically acceptable salt thereof.
[0080] In one group of selected embodiments, Ar is selected from
benzene, pyridine and quinoline, each of which is optionally
substituted with from one to two R.sup.3.
[0081] In another group of selected embodiments, L is selected from
the group consisting of a bond, --CH.sub.2-- and --CH(CH.sub.3)--.
In still another group of selected embodiments L.sup.2 is selected
from the group consisting of a bond, --O--CH.sub.2--,
--CH(CH.sub.3)--, --C(CH.sub.3).sub.2--, --CH.sub.2CH.sub.2--,
--CH.sub.2--, and --CH.sub.2CH.sub.2CH.sub.2--.
[0082] In other selected embodiments, suitable compounds are
selected from:
##STR00011##
or a pharmaceutically acceptable salt thereof, wherein said
compound is substantially free of other isomers.
[0083] Within formula (Ia1), (Ia2) and (Ia3), selected embodiments
are those wherein Ar is selected from the group consisting of
benzene, pyridine and quinoline, each of which is optionally
substituted with from one to two R.sup.3. In still other
embodiments, Ar is selected from the group consisting of
1,3-phenylene and 1,4-phenylene, each of which is optionally
substituted with from one to two R.sup.3. In some embodiments,
referring to formulae (Ia1), (Ia2) and (Ia3), L.sup.1 is selected
from the group consisting of a bond, --CH.sub.2-- and
--CH(CH.sub.3)--. In other embodiments of formulae (Ia1), (Ia2) and
(Ia3), L.sup.2 is selected from the group consisting of
--O--CH.sub.2--, --CH(CH.sub.3)--, --C(CH.sub.3).sub.2--,
--CH.sub.2CH.sub.2--, --CH.sub.2-- and
--CH.sub.2CH.sub.2CH.sub.2--. In still other embodiments of
formulae (Ia1), (Ia2) and (Ia3), R.sup.1 is selected from the group
consisting of halogen, cyano, C.sub.1-3 alkyl, C.sub.1-3 alkoxy,
C.sub.1-3 haloalkyl, C.sub.1-3 haloalkoxy, C.sub.3-5 cycloalkyl,
and C.sub.2-3 alkenyl. In yet other embodiments of formulae (Ia1),
(Ia2) and (Ia3), R.sup.1 is selected from the group consisting of
chloro, methyl, cyano, ethyl, cyclopropyl, trifluoromethyl and
trifluoromethoxy.
[0084] In other selected embodiments, suitable compounds are
selected from:
##STR00012##
or a pharmaceutically acceptable salt thereof, wherein said
compound is substantially free of other isomers.
[0085] In other embodiments, suitable compounds are provided having
the formula (Ib):
##STR00013##
or a pharmaceutically acceptable salt thereof.
[0086] In some embodiments of formula (Ib), R.sup.2b is hydrogen.
In other embodiments of formula (Ib), Ar is selected from the group
consisting of benzene, pyridine and quinoline, each of which is
optionally substituted with from one to two R.sup.3. In still other
embodiments of formula (Ib), L.sup.1 is selected from the group
consisting of a bond, --CH.sub.2-- and --CH(CH.sub.3)--. In yet
other embodiments of formula (Ib), L.sup.2 is selected from the
group consisting of a
bond, --O--CH.sub.2--, --CH(CH.sub.3)--, --C(CH.sub.3).sub.2--,
--CH.sub.2CH.sub.2--, --CH.sub.2-- and
--CH.sub.2CH.sub.2CH.sub.2--.
[0087] In other selected embodiments, suitable compounds are
selected from:
##STR00014##
or a pharmaceutically acceptable salt thereof, wherein said
compound is substantially free of other isomers.
[0088] Within formula (Ib1), (Ib2) and (Ib3), selected embodiments
are those wherein Ar is selected from the group consisting of
benzene, pyridine and quinoline, each of which is optionally
substituted with from one to two R.sup.3. In some embodiments,
referring to formulae (Ib1), (Ib2) and (Ib3), Ar is selected from
the group consisting of 1,3-phenylene and 1,4-phenylene, each of
which is optionally substituted with from one to two R.sup.3. In
other embodiments of formulae (Ib1), (Ib2) and (Ib3), R.sup.3 is
selected from the group consisting of CH.sub.3, CH.sub.2CH.sub.3,
CH.sub.2CH.sub.2CH.sub.3, CH(CH.sub.3).sub.2 and CH.sub.2OH. In
still other embodiments of formulae (Ib1), (Ib2) and (Ib3), L.sup.1
is selected from the group consisting of a bond, --CH.sub.2-- and
--CH(CH.sub.3)--. In yet other embodiments of formulae (Ib1), (Ib2)
and (Ib3), L.sup.2 is selected from the group consisting of
--O--CH.sub.2--, --CH(CH.sub.3)--, --C(CH.sub.3).sub.2--,
--CH.sub.2CH.sub.2--, --CH.sub.2-- and
--CH.sub.2CH.sub.2CH.sub.2--. In other embodiments of formulae
(Ib1), (Ib2) and (Ib3), R.sup.1 is selected from the group
consisting of halogen, cyano, C.sub.1-3 alkyl, C.sub.1-3 alkoxy,
C.sub.1-3 haloalkyl, C.sub.1-3 haloalkoxy, C.sub.3-5 cycloalkyl,
and C.sub.2-3 alkenyl, or R.sup.1 is selected from the group
consisting of chloro, methyl, cyano, ethyl, cyclopropyl,
trifluoromethyl and trifluoromethoxy.
[0089] In other selected embodiments, suitable compounds are
selected from:
##STR00015##
or a pharmaceutically acceptable salt thereof, wherein said
compound is substantially free of other isomers.
Preparation of Compounds
[0090] Compound provided herein can be prepared by the general
scheme below. Beginning with a suitably substituted phenylacetic
acid ester, reaction with a substituted halonitrobenzene in the
presence of base, following by methyl iodide sets the framework for
2-oxoindole ring construction having the quaternary center alpha to
the carboxylic acid. Resolution of the isomers, followed by
reduction of the nitro group and cyclization produces a substituted
2-oxoindole. Reaction at the indole nitrogen atom to attach either
a substituted Ar group, or a linker (L.sup.1), having an attached
substituted Ar group will lead to the target compounds shown. One
of skill in the art will appreciate that modifications can be made
following the general guidance of the scheme below to provide a
variety of compounds of formula (I).
##STR00016##
Compositions that Modulate Chemokine Activity
[0091] In another aspect, the present disclosure provides
compositions that modulate chemokine activity, specifically CCR(9)
activity. Generally, the compositions for modulating chemokine
receptor activity in humans and animals will comprise a
pharmaceutically acceptable excipient or diluent and a compound
having any of the formulae I, I', Ia, Ib, Ia1, Ia2, Ia3, Ia1',
Ia2', Ia3', Ib1, Ib2, Ib3, Ib1', Ib2' and Ib3'.
[0092] The term "composition" as used herein is intended to
encompass a product comprising the specified ingredients in the
specified amounts, as well as any product which results, directly
or indirectly, from combination of the specified ingredients in the
specified amounts. By "pharmaceutically acceptable" it is meant the
carrier, diluent or excipient must be compatible with the other
ingredients of the formulation and not deleterious to the recipient
thereof.
[0093] The pharmaceutical compositions for the administration of
the compounds of this disclosure may conveniently be presented in
unit dosage form and may be prepared by any of the methods well
known in the art of pharmacy. All methods include the step of
bringing the active ingredient into association with the carrier
which constitutes one or more accessory ingredients. In general,
the pharmaceutical compositions are prepared by uniformly and
intimately bringing the active ingredient into association with a
liquid carrier or a finely divided solid carrier or both, and then,
if necessary, shaping the product into the desired formulation. In
the pharmaceutical composition the active object compound is
included in an amount sufficient to produce the desired effect upon
the process or condition of diseases.
[0094] The pharmaceutical compositions containing the active
ingredient may be in a form suitable for oral use, for example, as
tablets, troches, lozenges, aqueous or oily suspensions,
dispersible powders or granules, emulsions and self-emulsifications
as described in U.S. Pat. No. 6,451,339, hard or soft capsules, or
syrups or elixirs. Compositions intended for oral use may be
prepared according to any method known to the art for the
manufacture of pharmaceutical compositions. Such compositions may
contain one or more agents selected from sweetening agents,
flavoring agents, coloring agents and preserving agents in order to
provide pharmaceutically elegant and palatable preparations.
Tablets contain the active ingredient in admixture with other
non-toxic pharmaceutically acceptable excipients which are suitable
for the manufacture of tablets. These excipients may be, for
example, inert diluents such as cellulose, silicon dioxide,
aluminum oxide, calcium carbonate, sodium carbonate, glucose,
mannitol, sorbitol, lactose, calcium phosphate or sodium phosphate;
granulating and disintegrating agents, for example, corn starch, or
alginic acid; binding agents, for example PVP, cellulose, PEG,
starch, gelatin or acacia, and lubricating agents, for example
magnesium stearate, stearic acid or talc. The tablets may be
uncoated or they may be coated enterically or otherwise by known
techniques to delay disintegration and absorption in the
gastrointestinal tract and thereby provide a sustained action over
a longer period. For example, a time delay material such as
glyceryl monostearate or glyceryl distearate may be employed. They
may also be coated by the techniques described in the U.S. Pat.
Nos. 4,256,108; 4,166,452; and U.S. Pat. No. 4,265,874 to form
osmotic therapeutic tablets for control release.
[0095] Formulations for oral use may also be presented as hard
gelatin capsules wherein the active ingredient is mixed with an
inert solid diluent, for example, calcium carbonate, calcium
phosphate or kaolin, or as soft gelatin capsules wherein the active
ingredient is mixed with water or an oil medium, for example peanut
oil, liquid paraffin, or olive oil. Additionally, emulsions can be
prepared with a non-water miscible ingredient such as oils and
stabilized with surfactants such as mono-diglycerides, PEG esters
and the like.
[0096] Aqueous suspensions contain the active materials in
admixture with excipients suitable for the manufacture of aqueous
suspensions. Such excipients are suspending agents, for example
sodium carboxymethylcellulose, methylcellulose,
hydroxypropylmethylcellulose, sodium alginate,
polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or
wetting agents may be a naturally-occurring phosphatide, for
example lecithin, or condensation products of an alkylene oxide
with fatty acids, for example polyoxyethylene stearate, or
condensation products of ethylene oxide with long chain aliphatic
alcohols, for example heptadecaethyleneoxycetanol, or condensation
products of ethylene oxide with partial esters derived from fatty
acids and a hexitol such as polyoxyethylene sorbitol monooleate, or
condensation products of ethylene oxide with partial esters derived
from fatty acids and hexitol anhydrides, for example polyethylene
sorbitan monooleate. The aqueous suspensions may also contain one
or more preservatives, for example ethyl, or n-propyl,
p-hydroxybenzoate, one or more coloring agents, one or more
flavoring agents, and one or more sweetening agents, such as
sucrose or saccharin.
[0097] Oily suspensions may be formulated by suspending the active
ingredient in a vegetable oil, for example arachis oil, olive oil,
sesame oil or coconut oil, or in a mineral oil such as liquid
paraffin. The oily suspensions may contain a thickening agent, for
example beeswax, hard paraffin or cetyl alcohol. Sweetening agents
such as those set forth above, and flavoring agents may be added to
provide a palatable oral preparation. These compositions may be
preserved by the addition of an anti-oxidant such as ascorbic
acid.
[0098] Dispersible powders and granules suitable for preparation of
an aqueous suspension by the addition of water provide the active
ingredient in admixture with a dispersing or wetting agent,
suspending agent and one or more preservatives. Suitable dispersing
or wetting agents and suspending agents are exemplified by those
already mentioned above. Additional excipients, for example
sweetening, flavoring and coloring agents, may also be present.
[0099] The pharmaceutical compositions of the disclosure may also
be in the form of oil in water emulsions. The oily phase may be a
vegetable oil, for example olive oil or arachis oil, or a mineral
oil, for example liquid paraffin or mixtures of these. Suitable
emulsifying agents may be naturally-occurring gums, for example gum
acacia or gum tragacanth, naturally-occurring phosphatides, for
example soy bean, lecithin, and esters or partial esters derived
from fatty acids and hexitol anhydrides, for example sorbitan
monooleate, and condensation products of the said partial esters
with ethylene oxide, for example polyoxyethylene sorbitan
monooleate. The emulsions may also contain sweetening and flavoring
agents.
[0100] Syrups and elixirs may be formulated with sweetening agents,
for example glycerol, propylene glycol, sorbitol or sucrose. Such
formulations may also contain a demulcent, a preservative, and
flavoring and coloring agents. Oral solutions can be prepared in
combination with, for example, cyclodextrin, PEG and
surfactants.
[0101] The pharmaceutical compositions may be in the form of a
sterile injectable aqueous or oleaginous suspension. This
suspension may be formulated according to the known art using those
suitable dispersing or wetting agents and suspending agents which
have been mentioned above. The sterile injectable preparation may
also be a sterile injectable solution or suspension in a non-toxic
parenterally acceptable diluent or solvent, for example as a
solution in 1,3-butane diol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution and
isotonic sodium chloride solution. In addition, sterile, axed oils
are conventionally employed as a solvent or suspending medium. For
this purpose any bland fixed oil may be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid find use in the preparation of injectables.
[0102] The compounds of the present disclosure may also be
administered in the form of suppositories for rectal administration
of the drug. These compositions can be prepared by mixing the drug
with a suitable non-irritating excipient which is solid at ordinary
temperatures but liquid at the rectal temperature and will
therefore melt in the rectum to release the drug. Such materials
are cocoa butter and polyethylene glycols. Additionally, the
compounds can be administered via ocular delivery by means of
solutions or ointments. Still further, transdermal delivery of the
subject compounds can be accomplished by means of iontophoretic
patches and the like.
[0103] For topical use, creams, ointments, jellies, solutions or
suspensions containing the compounds of the present disclosure are
employed. As used herein, topical application is also meant to
include the use of mouth washes and gargles.
[0104] The pharmaceutical compositions and methods of the present
disclosure may further comprise other therapeutically active
compounds as noted herein, such as those applied in the treatment
of the above mentioned pathological conditions.
[0105] In one embodiment, the present disclosure provides a
composition consisting of a pharmaceutically acceptable carrier and
a compound of the disclosure.
Methods of Treatment
[0106] Depending on the disease to be treated and the subject's
condition, the compounds and compositions of the present disclosure
may be administered by oral, parenteral (e.g., intramuscular,
intraperitoneal, intravenous, ICV, intracisternal injection or
infusion, subcutaneous injection, or implant), inhalation, nasal,
vaginal, rectal, sublingual, or topical routes of administration
and may be formulated, alone or together, in suitable dosage unit
formulations containing conventional non-toxic pharmaceutically
acceptable carriers, adjuvants and vehicles appropriate for each
route of administration. The present disclosure also contemplates
administration of the compounds and compositions of the present
disclosure in a depot formulation.
[0107] In the treatment or prevention of conditions which require
chemokine receptor modulation an appropriate dosage level will
generally be about 0.001 to 100 mg per kg patient body weight per
day which can be administered in single or multiple doses.
Preferably, the dosage level will be about 0.01 to about 25 mg/kg
per day; more preferably about 0.05 to about 10 mg/kg per day. A
suitable dosage level may be about 0.01 to 25 mg/kg per day, about
0.05 to 10 mg/kg per day, or about 0.1 to 5 mg/kg per day. Within
this range the dosage may be 0.005 to 0.05, 0.05 to 0.5, 0.5 to
5.0, or 5.0 to 50 mg/kg per day. For oral administration, the
compositions are preferably provided in the form of tablets
containing 1.0 to 1000 milligrams of the active ingredient,
particularly 1.0, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0,
150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0,
900.0, and 1000.0 milligrams of the active ingredient for the
symptomatic adjustment of the dosage to the patient to be treated.
The compounds may be administered on a regimen of 1 to 4 times per
day, preferably once or twice per day.
[0108] It will be understood, however, that the specific dose level
and frequency of dosage for any particular patient may be varied
and will depend upon a variety of factors including the activity of
the specific compound employed, the metabolic stability and length
of action of that compound, the age, body weight, hereditary
characteristics, general health, sex, diet, mode and time of
administration, rate of excretion, drug combination, the severity
of the particular condition, and the host undergoing therapy.
[0109] In some embodiments, compounds of the present disclosure are
administered as part of a combination therapy. For instance an
amount of a chemotherapeutic agent or radiation is administered to
the subject prior to, subsequent to or in combination with the
compounds of the present disclosure. In some embodiments, the
amount is sub-therapeutic when the chemotherapeutic agent or
radiation is administered alone. Those of skill in the art will
appreciate that "combinations" can involve combinations in
treatments (i.e., two or more drugs can be administered as a
mixture, or at least concurrently or at least introduced into a
subject at different times but such that both are in the
bloodstream of a subject at the same time). Additionally,
compositions of the current disclosure may be administered prior to
or subsequent to a second therapeutic regimen, for instance prior
to or subsequent to a dose of chemotherapy or irradiation.
[0110] In still other embodiments, the present methods are directed
to the treatment of allergic diseases, wherein a compound or
composition of the disclosure is administered either alone or in
combination with a second therapeutic agent, wherein said second
therapeutic agent is an antihistamine or an anti-inflammatory. When
used in combination, the practitioner can administer a combination
of the compound or composition of the present disclosure and a
second therapeutic agent. Also, the compound or composition and the
second therapeutic agent can be administered sequentially, in any
order.
[0111] The compounds and compositions of the present disclosure can
be combined with other compounds and compositions having related
utilities to prevent and treat the condition or disease of
interest, such as inflammatory conditions and diseases, including
inflammatory bowel disease (including Crohn's disease and
ulcerative colitis), allergic diseases, psoriasis, atopic
dermatitis and asthma, and those pathologies noted above. Selection
of the appropriate agents for use in combination therapies can be
made one of ordinary skill in the art. The combination of
therapeutic agents may act synergistically to effect the treatment
or prevention of the various disorders. Using this approach, one
may be able to achieve therapeutic efficacy with lower dosages of
each agent, thus reducing the potential for adverse side
effects.
[0112] In treating, preventing, ameliorating, controlling or
reducing the risk of inflammation, the compounds of the present
disclosure may be used in conjunction with an antiinflammatory or
analgesic agent such as an opiate agonist, a lipoxygenase
inhibitor, such as an inhibitor of 5-lipoxygenase, a cyclooxygenase
inhibitor, such as a cyclooxygenase-2 inhibitor, an interleukin
inhibitor, such as an interleukin-1 inhibitor, an NMDA antagonist,
an inhibitor of nitric oxide or an inhibitor of the synthesis of
nitric oxide, aminosalicylates, corticosteroids and other
immunosuppressive drugs, a non-steroidal antiinflammatory agent, or
a cytokine-suppressing antiinflammatory agent, for example with a
compound such as acetaminophen, aspirin, codeine, biological TNF
sequestrants, biological agents which target .alpha.4.beta.7, ACE2
inhibitors, protein linase C inhibitors, fentanyl, ibuprofen,
indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam,
a steroidal analgesic, sufentanyl, sunlindac, tenidap, and the
like.
[0113] Similarly, the compounds of the present disclosure may be
administered with a pain reliever; a potentiator such as caffeine,
an H2-antagonist, simethicone, aluminum or magnesium hydroxide; a
decongestant such as pseudophedrine; an antitussive such as
codeine; a diuretic; a sedating or non-sedating antihistamine; a
very late antigen (VLA-4) antagonist; an immunosuppressant such as
cyclosporin, tacrolimus, rapamycin, EDG receptor agonists, or other
FK-506 type immunosuppressants; a steroid; a non-steroidal
anti-asthmatic agent such as a .beta.2-agonist, leukotriene
antagonist, or leukotriene biosynthesis inhibitor; an inhibitor of
phosphodiesterase type IV (PDE-IV); a cholesterol lowering agent
such as a HMG-CoA reductase inhibitor, sequestrant, or cholesterol
absorption inhibitor; and an anti-diabetic agent such as insulin,
.alpha.-glucosidase inhibitors or glitazones.
[0114] The weight ratio of the compound of the present disclosure
to the second active ingredient may be varied and will depend upon
the effective dose of each ingredient. Generally, an effective dose
of each will be used. Thus, for example, when a compound of the
present disclosure is combined with an NSAID the weight ratio of
the compound of the present disclosure to the NSAID will generally
range from about 1000:1 to about 1:1000, preferably about 200:1 to
about 1:200. Combinations of a compound of the present disclosure
and other active ingredients will generally also be within the
aforementioned range, but in each case, an effective dose of each
active ingredient should be used.
Methods of Treating or Preventing CCR(9)-Mediated Conditions or
Diseases
[0115] In yet another aspect, the present disclosure provides
methods of treating or preventing a CCR(9)-mediated condition or
disease by administering to a subject having such a condition or
disease a therapeutically effective amount of any compound of
formula I, I', Ia, Ib, Ia1, Ia2, Ia3, Ia1', Ia2', Ia3', Ib1, Ib2,
Ib3, Ib1', Ib2' or Ib3'. Compounds for use in the present methods
include those compounds according to the formula I, I', Ia, Ib,
Ia1, Ia2, Ia3, Ia1', Ia2', Ia3', Ib1, Ib2, Ib3, Ib1', Ib2' and
Ib3', those provided above as embodiments, those specifically
exemplified in the Examples below, and those provided with specific
structures herein. The "subject" is defined herein to include
animals such as mammals, including, but not limited to, primates
(e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits,
rats, mice and the like. In preferred embodiments, the subject is a
human.
[0116] As used herein, the phrase "CCR(9)-mediated condition or
disease" and related phrases and terms refer to a condition or
disease characterized by inappropriate, i.e., less than or greater
than normal, CCR(9) functional activity. Inappropriate CCR(9)
functional activity might arise as the result of CCR(9) expression
in cells which normally do not express CCR(9), increased CCR(9)
expression (leading to, e.g., inflammatory and immunoregulatory
disorders and diseases) or decreased CCR(9) expression.
Inappropriate CCR(9) functional activity might also arise as the
result of TECK secretion by cells which normally do not secrete
TECK, increased TECK expression (leading to, e.g., inflammatory and
immunoregulatory disorders and diseases) or decreased TECK
expression. A CCR(9)-mediated condition or disease may be
completely or partially mediated by inappropriate CCR(9) functional
activity. However, a CCR(9)-mediated condition or disease is one in
which modulation of CCR(9) results in some effect on the underlying
condition or disease (e.g., a CCR(9) antagonist results in some
improvement in patient well-being in at least some patients).
[0117] The term "therapeutically effective amount" means the amount
of the subject compound that will elicit the biological or medical
response of a cell, tissue, system, or animal, such as a human,
that is being sought by the researcher, veterinarian, medical
doctor or other treatment provider.
[0118] Diseases and conditions associated with inflammation, immune
disorders, infection and cancer may be treated or prevented with
the present compounds, compositions, and methods. In one group of
embodiments, diseases or conditions, including chronic diseases, of
humans or other species maybe treated with inhibitors of CCR(9)
function. These diseases or conditions include: (1) allergic
diseases such as systemic anaphylaxis or hypersensitivity
responses, drug allergies, insect sting allergies and food
allergies, (2) inflammatory bowel diseases, such as Crohn's
disease, ulcerative colitis, microscopic colitis, ileitis and
enteritis, and postoperative ileus, (3) vaginitis, (4) psoriasis
and inflammatory dermatoses such as dermatitis, eczema, atopic
dermatitis, allergic contact dermatitis, urticaria and pruritus,
(5) vasculitis, (6) spondyloarthropathies, (7) scleroderma, (8)
asthma and respiratory allergic diseases such as allergic asthma,
allergic rhinitis, hypersensitivity lung diseases and the like, (9)
autoimmune diseases, such as fibromyalagia, ankylosing spondylitis,
juvenile RA, Still's disease, polyarticular juvenile RA,
pauciarticular juvenile RA, polymyalgia rheumatica, rheumatoid
arthritis, psoriatic arthritis, osteoarthritis, polyarticular
arthritis, multiple sclerosis, systemic lupus erythematosus, type I
diabetes, type II diabetes, glomerulonephritis, and the like, (10)
graft rejection (including allograft rejection), (11) graft-v-host
disease (including both acute and chronic), (12) other diseases in
which undesired inflammatory responses are to be inhibited, such as
atherosclerosis, myositis, neurodegenerative diseases (e.g.,
Alzheimer's disease), encephalitis, meningitis, hepatitis,
nephritis, sepsis, sarcoidosis, allergic conjunctivitis, otitis,
chronic obstructive pulmonary disease, sinusitis, Behcet's syndrome
and gout, (13) immune mediated food allergies such as Coeliac
(Celiac) disease (14) pulmonary fibrosis and other fibrotic
diseases, (15) irritable bowel syndrome, (16) primary sclerosing
cholangitis, (17) cancer (including both primary and metastatic),
(18) bacterial associated syndromes such as hemorrhagic colitis and
hemolytic uremic syndrome (19) melanoma, (20) primary sclerosing
cholangitis, (21) post-operative ileus (22) hepatitis and
inflammatory hepatic diseases (23) Sjogren syndrome.
[0119] In another group of embodiments, diseases or conditions may
be treated with modulators and agonists of CCR(9) function.
Examples of diseases that may be treated by modulating CCR(9)
function include cancers, cardiovascular diseases, diseases in
which angiogenesis or neovascularization play a role (neoplastic
diseases, retinopathy and macular degeneration), infectious
diseases (viral infections, e.g., HIV infection, and bacterial
infections) and immunosuppressive diseases such as organ transplant
conditions and skin transplant conditions. The term "organ
transplant conditions" is means to include bone marrow transplant
conditions and solid organ (e.g., kidney, liver, lung, heart,
pancreas or combination thereof) transplant conditions.
[0120] Preferably, the present methods are directed to the
treatment of diseases or conditions selected from inflammatory
bowel disease including Crohn's disease and Ulcerative Colitis,
allergic diseases, psoriasis, atopic dermatitis and asthma,
autoimmune disease such as rheumatoid arthritis and immune-mediated
food allergies such as Celiac disease.
[0121] In yet other embodiments, the present methods are directed
to the treatment of psoriasis where a compound or composition of
the disclosure is used alone or in combination with a second
therapeutic agent such as a corticosteroid, a lubricant, a
keratolytic agent, a vitamin D.sub.3 derivative, PUVA and
anthralin.
[0122] In other embodiments, the present methods are directed to
the treatment of atopic dermatitis using a compound or composition
of the disclosure either alone or in combination with a second
therapeutic agent such as a lubricant and a corticosteroid.
[0123] In further embodiments, the present methods are directed to
the treatment of asthma using a compound or composition of the
disclosure either alone or in combination with a second therapeutic
agent such as a .beta.2-agonist and a corticosteroid.
Kits and Packages
[0124] The terms "kit" and "pharmaceutical kit" refer to a
commercial kit or package comprising, in one or more suitable
containers, one or more pharmaceutical compositions and
instructions for their use. In one embodiment, kits comprising a
compound of Formula I, I', Ia, Ib, Ia1, Ia2, Ia3, Ia1', Ia2', Ia3',
Ib1, Ib2, Ib3, Ib1', Ib2' or Ib3', or a pharmaceutically acceptable
salt thereof, and instructions for its administration are provided.
In one embodiment, kits comprising a compound of Formula I, I', Ia,
Ib, Ia1, Ia2, Ia3, Ia1', Ia2', Ia3', Ib1, Ib2, Ib3, Ib1', Ib2' or
Ib3', or a pharmaceutically acceptable salt thereof, in combination
with one or more (e.g., one, two, three, one or two, or one to
three) additional therapeutic agents and instructions for their
administration are provided.
[0125] In one embodiment, the compounds of this disclosure are
formulated into administration units which are packaged in a single
packaging. The single packaging encompasses but is not limited to a
bottle, a child-resistant bottle, an ampoule, and a tube. In one
embodiment, the compounds of this disclosure and optionally
additional therapeutic agents, are formulated into administration
units and every single administration unit is individually packaged
in a single packaging. Such individually packaged units may contain
the pharmaceutical composition in any form including but not
limited to liquid form, solid form, powder form, granulate form, an
effervescent powder or tablet, hard or soft capsules, emulsions,
suspensions, syrup, suppositories, tablet, troches, lozenges,
solution, buccal patch, thin film, oral gel, chewable tablet,
chewing gum, and single-use syringes. Such individually packaged
units may be combined in a package made of one or more of paper,
cardboard, paperboard, metal foil and plastic foil, for example a
blister pack. One or more administration units may be administered
once or several times a day. One or more administration units may
be administered three times a day. One or more administration units
may be administered twice a day. One or more administration units
may be administered on a first day and one or more administration
units may be administered on the following days.
Additional Combination Therapies
[0126] The compounds of this disclosure can be supplied alone or in
conjunction with one or more other drugs. Examples of therapeutic
agents that may be combined with a compound or composition of the
present disclosure, either administered separately or in the same
pharmaceutical composition, include, but are not limited to:
modulators of CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9,
CCR10, CCR11, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, CXCR7,
CX3CR1, ChemR23, C5aR, C5a, and C5, or any combination thereof. In
some embodiments, the modulator is an antagonist.
[0127] Examples of therapeutic agents that may be combined with a
compound or composition of the present disclosure, either
administered separately or in the same pharmaceutical composition,
include, but are not limited to: CCX354, CCX9588, CCX140, CCX872,
CCX598, CCX6239, CCX9664, CCX2553, CCX 2991, CCX282, CCX025,
CCX507, CCX430, CCX765, CCX224, CCX662, CCX650, CCX832, CCX168,
CCX168-M1, brazikumab, budesonide, ustekinumab, everolimus,
glatiramer acetate, natalizumab, etanercept, mycophenolate mofetil,
brodalumab, cannabidiol, foralumab, tralokinumab, tamibarotene,
mesalazine, golimumab, teduglutide, infliximab, ropivacaine,
filgotinib, etrolizumab, SHP-647, elafibranor, ABC-294640,
ocrelizumab, tofacitinib, certolizumab pegol, adalimumab,
sargramostim, abatacept, clarithromycin, GSK-2982772, upadacitinib,
edasalonexent, secukinumab, vancomycin, vedolizumab, thalidomide,
rituximab, catridecacog, RBX-2660, Ampion, nitazoxanide,
fingolimod, tocilizumab, rosiptor acetate, AST-120, risankizumab,
telotristat etiprate, lenalidomide, alicaforsen, tosufloxacin,
interferon beta-1a, E-6011, KAG-308, dexamethasone sodium
phosphate, ozanimod, dociparstat cobitolimod, mesalazine, PUR-0110,
apremilast, mesalazine, valganciclovir, tacrolimus, mongersen,
remestemcel-L, GS-5745, E-6011, E-6007, carotegrast methyl,
piclidenoson, PF-06480605, balsalazide, pimecrolimus, mesalazine,
recombinant interferon beta-1a, naltrexone, adalimumab, amiselimod,
brilacidin, basiliximab, etrasimod, LP-02, rosiglitazone,
plecanatide, laquinimod, rifabutin+clarithromycin+clofazimine,
infliximab, tildrakizumab, omega-3-carboxylic acids, TOP-1288,
peficitinib, rifamycin, rifaximin, JNJ-64304500, ASP-3291, DLX-105,
zileuton, 99mTc labelled annexin V-128, ALT-836, Biferonex,
clotrimazole, givinostat, Trichuris suis ova, INV-103, K(D)PT,
BI-655064, glepaglutide, LYC-30937 EC, TRX-318, LY-3074828,
larazotide acetate, IBP-9414, clazakizumab, mesalazine,
eclomethasone dipropionate, NN-8828, olokizumab, bertilimumab,
midismase, KRP-203, prednisolone, PF-06687234, STNM-01, KHK-4083,
FE-999301, DLX-105, VB-201, DNVX-078, rifaximin, Clostridium
butyricum MIYAIRI 588, OPS-2071, sotrastaurin, abrilumab, QBECO,
anakinra, FFP-104, GLPG-1205, dolcanatide, PDA-002, molgramostim,
mesalazine, metronidazole, repurposed naltrexone, vatelizumab,
zucapsaicin, ciclosporin, oprelvekin, prulifloxacin, recombinant
human lactoferrin, Alequel, SAN-300, STP-206, GLPG-0974, P-28-GST,
N-6022, TNF alpha kinoid, ETX-201, low molecular weight heparin,
ETX-201, GED-0507-34-Levo, metenkefalin acetate+tridecactide
acetate, HMPL-004, SB-012, TRK-170, beta-1,3/1,6-glucan,
mesalamine+N-acetylcysteine, 99mTc-sulesomab, olsalazine,
mesalazine bacillus licheniformis, balsalazide sodium,
propionyl-L-camitine, Clostridium butyricum, beclomethasone
dipropionate, acemannan, and SPD-480, or any combination
thereof.
[0128] Examples of therapeutic agents that may be combined with a
compound or composition of the present disclosure, either
administered separately or in the same pharmaceutical composition,
include, but are not limited to: an IL-23 antagonist, a
Glucocorticoid agonist, an IL-6 agonist, an IL-12 antagonist, a
mTOR complex 1 inhibitor, a mTOR inhibitor, a cell adhesion
molecule inhibitor, an Integrin alpha-4/beta-1 antagonist, a TNF
antagonist, a TNF binding agent, a Type II TNF receptor modulator,
an Inosine monophosphate dehydrogenase inhibitor, a PurH purine
biosynthesis protein inhibitor, an Interleukin receptor 17A,
antagonist, a Cannabinoid CB1 receptor modulator, a Cannabinoid CB2
receptor modulator, a Cannabinoid receptor modulator, a CD3
antagonist, an IL-13 antagonist, a Retinoic acid receptor alpha
agonist, a Retinoic acid receptor beta agonist, a Retinoid receptor
agonist, a Cyclooxygenase inhibitor, a TNF alpha ligand inhibitor,
a Glucagon-like peptide 2 agonist, a sodium channel inhibitor, a
Jak1 tyrosine kinase inhibitor, an Integrin alpha-4/beta-7
antagonist, an Integrin alpha-E antagonist, an Integrin beta-7
antagonist, an Immunoglobulin G2 modulator, a MAdCAM inhibitor, an
Insulin sensitizer, a PPAR alpha agonist, a PPAR delta agonist, a
Collagen modulator, a Dihydroceramide delta 4 desaturase inhibitor,
a Sphingosine kinase 1 inhibitor, a Sphingosine kinase 2 inhibitor,
aB-lymphocyte antigen CD20 inhibitor, a JAK tyrosine kinase
inhibitor, a Jak3 tyrosine kinase inhibitor, a CSF-1 agonist, a
GM-CSF receptor agonist, a Cytotoxic T-lymphocyte protein-4
stimulator, a T cell surface glycoprotein CD28 inhibitor, a RIP-1
kinase inhibitor, a Nuclear factor kappa B inhibitor, an IL-17
antagonist, a Peptidoglycan recognition protein inhibitor, an
Integrin alpha-4/beta-7 antagonist, a B-lymphocyte antigen CD20
inhibitor, a Factor XIII agonist, a Stem cell antigen-1 inhibitor,
a Cannabinoid receptor antagonist; Sphingosine-1-phosphate
receptor-1 modulator, an IL-6 antagonist, an IL-6 receptor
modulator, a SH2 domain inositol phosphatase 1 stimulator, a
Tryptophan 5-hydroxylase inhibitor, a ICAM1 gene inhibitor, a DNA
gyrase inhibitor, a Topoisomerase IV inhibitor, an Interferon beta
ligand, a Fractalkine ligand inhibitor, a EP4 prostanoid receptor
agonist, a Sphingosine-1-phosphate receptor-1 agonist, a
Sphingosine-1-phosphate receptor-1 modulator, a
Sphingosine-1-phosphate receptor-5 modulator, a Cathepsin G
inhibitor, a Complement cascade inhibitor, an Elastase inhibitor, a
Heparin agonist, a L-Selectin antagonist, a P-Selectin antagonist,
a Nuclear factor kappa B inhibitor, a TLR-9 agonist, an
Interleukin-1 beta ligand modulator, a PDE 4 inhibitor, a DNA
polymerase inhibitor, a SMAD-7 inhibitor, a TGF beta 1 ligand
inhibitor, a Metalloprotease-9 inhibitor, a Fractalkine ligand
inhibitor, an Integrin antagonist, an Adenosine A3 receptor
agonist, a Tumor necrosis factor 15 ligand inhibitor, an IL-10
antagonist, an IL-2 antagonist, an IL-4 antagonist, an Interferon
gamma receptor antagonist, an Interferon beta ligand, an Opioid
receptor antagonist, an IL-2 receptor alpha subunit inhibitor, a
Sphingosine 1 phosphate phosphatase 1 stimulator, an Insulin
sensitizer, a PPAR gamma agonist, a Natriuretic peptide receptor C
agonist, a n acyltransferase inhibitor, an apolipoprotein C3
antagonist, an adapter molecule crk inhibitor, an IL-8 antagonist,
an Interleukin-1 beta ligand inhibitor, a Src tyrosine kinase
inhibitor, a Syk tyrosine kinase inhibitor, a DNA RNA polymerase
inhibitor, a RNA polymerase inhibitor, a Melanocortin agonist, a
5-Lipoxygenase inhibitor, a Tissue factor inhibitor, an Interferon
beta ligand, a Bradykinin receptor modulator, an Histone
deacetylase inhibitor, a P2X7 purinoceptor agonist, a mitochondrial
10 kDa heat shock protein stimulator, a CD40 ligand receptor
antagonist, a Glucagon-like peptide 2 agonist, a FIFO ATP synthase
modulator, a CD3 antagonist, a Zonulin inhibitor, a Cyclooxygenase
inhibitor, a Lipoxygenase modulator, an IL-21 antagonist, a CCR3
chemokine antagonist, an Eotaxin ligand inhibitor, a Superoxide
dismutase modulator, a Sphingosine-1-phosphate receptor-1 agonist,
a CD29 modulator, an Interleukin-10 ligand, a CHST15 gene
inhibitor, an OX40 ligand inhibitor, an IL-6 receptor modulator, a
Nuclear factor kappa B inhibitor, an Oncostatin M receptor
modulator, a STAT inhibitor, a STAT-3 inhibitor, a TLR-2
antagonist, a TLR-4 antagonist, a RNA polymerase inhibitor, a
Protein kinase C alpha inhibitor, a Protein kinase C beta
inhibitor, a Protein kinase C delta inhibitor, a Protein kinase C
epsilon inhibitor, a Protein kinase C eta inhibitor, a Protein
kinase C theta inhibitor, a Type I IL-1 receptor antagonist, a CD40
ligand inhibitor, a CD40 ligand receptor antagonist, a G-protein
coupled receptor 84 antagonist, a Guanylate cyclase receptor
agonist, a CD49b antagonist, a Vanilloid VR1 agonist, a Calcineurin
inhibitor, an IL-11 agonist, a PDGF receptor agonist, a DNA gyrase
inhibitor, a Lactoferrin stimulator, an Integrin alpha-1/beta-1
antagonist, a Free fatty acid receptor 2 antagonist, an Alcohol
dehydrogenase 5 inhibitor, a glutathione reductase inhibitor, an
Interferon gamma receptor antagonist, Low molecular weight heparin,
a PPAR gamma agonist, a ACTH receptor agonist, an
Adrenocorticotrophic hormone ligand, an Opioid growth factor
receptor agonist, an IL-6 antagonist, an Interleukin-1 beta ligand
modulator, a Nuclear factor kappa B inhibitor; a GATA 3
transcription factor inhibitor, a Nuclear factor kappa B inhibitor,
an Oxidoreductase inhibitor, a Glucocorticoid agonist, an
Interferon gamma receptor agonist, or any combination thereof.
Examples
[0129] The following examples are offered to illustrate, but not to
limit the claimed disclosure.
[0130] Reagents and solvents used below can be obtained from
commercial sources such as Aldrich Chemical Co. (Milwaukee, Wis.,
USA). .sup.1H-NMR were recorded on a Varian Mercury 400 MHz NMR
spectrometer. Significant peaks are provided relative to TMS and
are tabulated in the order: multiplicity (s, singlet; d, doublet;
t, triplet; q, quartet; m, multiplet) and number of protons. Mass
spectrometry results are reported as the ratio of mass over charge,
followed by the relative abundance of each ion (in parenthesis). In
tables, a single m/e value is reported for the M+H (or, as noted,
M-H) ion containing the most common atomic isotopes. Isotope
patterns correspond to the expected formula in all cases.
Electrospray ionization (ESI) mass spectrometry analysis was
conducted on a Hewlett-Packard MSD electrospray mass spectrometer
using the HP 1100 HPLC equipped with an Agilent Zorbax SB-C18,
2.1.times.50 mm, 5.mu. column for sample delivery. Normally the
analyte was dissolved in methanol at 0.1 mg/mL and 1 microlitre was
infused with the delivery solvent into the mass spectrometer, which
scanned from 100 to 1500 daltons. All compounds could be analyzed
in the positive ESI mode, using acetonitrile/water with 1% formic
acid as the delivery solvent. The compounds provided below could
also be analyzed in the negative ESI mode, using 2 mM NH.sub.4OAc
in acetonitrile/water as delivery system.
[0131] The following abbreviations are used in the Examples and
throughout the description of the disclosure: [0132] HPLC, High
Pressure Liquid Chromatography; DMF, Dimethyl formamide; TFA,
Trifluoroacetic Acid; THF, Tetrahydrofuran; EtOAc, Ethyl acetate;
BOC.sub.2O, di-tertbutyl dicarbonate or BOC anhydride; HPLC, High
Pressure Liquid Chromatography; DIPEA, Diisopropyl ethylamine;
HBTU, O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate; dppf, 1,1'-Bis(diphenylphosphino)ferrocene;
Pd.sub.2(dba).sub.3, Tris(dibenzylideneacetone)dipalladium(0);
DIPEA, diisopropylethylamine; DMP, dimethylphthalate; Me, methyl;
Et, ethyl; DCM, dichloromethane.
[0133] Compounds within the scope of this disclosure can be
synthesized as described below, using a variety of reactions known
to the skilled artisan. One skilled in the art will also recognize
that alternative methods may be employed to synthesize the target
compounds of this disclosure, and that the approaches described
within the body of this document are not exhaustive, but do provide
broadly applicable and practical routes to compounds of
interest.
[0134] Certain molecules disclosed in this patent can exist in
different enantiomeric and diastereomeric forms and all such
variants of these compounds are claimed.
[0135] The detailed description of the experimental procedures used
to synthesize key compounds in this text lead to molecules that are
described by the physical data identifying them as well as by the
structural depictions associated with them.
[0136] Those skilled in the art will also recognize that during
standard work up procedures in organic chemistry, acids and bases
are frequently used. Salts of the parent compounds are sometimes
produced, if they possess the necessary intrinsic acidity or
basicity, during the experimental procedures described within this
patent.
Example 1: Synthesis of
(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one
##STR00017##
[0138] a) A solution of silver triflate (0.385 g, 1.50 mmol) in
1,2-dichloroethane (1.0 L) was heated at 90.degree. C. for 3 h
under a reflux condenser and nitrogen atmosphere. The cloudy
mixture was cooled to room temperature before methyl
4-hydroxyphenylacetate (24.9 g, 150 mmol) was added. Isoprene (15.3
g, 225 mmol) in 1,2-dichloroethane (100 mL) was added dropwise over
10 min and the mixture was stirred at room temperature for 2 h. The
solution was concentrated, diluted in EtOAc (150 mL), and washed
with saturated aqueous NaHCO.sub.3 (2.lamda.100 mL) and 1 M
NaHSO.sub.4 (2.times.100 mL). The organic layer was dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. Purification
of the crude material by flash chromatography (1-10% EtOAc in
hexanes) eluted methyl 2-(2,2-dimethylchroman-6-yl)acetate.
[0139] b) To a cooled (-30.degree. C.) solution of sodium
tert-butoxide (93.0 g, 963 mmol) in anhydrous N-methylpyrrolidone
(500 mL) under nitrogen atmosphere was slowly added methyl
2-(2,2-dimethylchroman-6-yl)acetate (80.8 g, 344 mmol), followed by
4-chloro-2-fluoronitrobenzene (62.1 g, 354 mmol), and more
N-methylpyrrolidone (150 mL). After stirring at -30.degree. C. for
1 h, methyl iodide (42.8 mL, 688 mmol) was added by syringe and the
mixture was stirred for an additional 20 min. The reaction was
quenched by addition of 3 M HCl (600 mL) and the mixture was warmed
to room temperature. The reaction mixture was extracted with methyl
tert-butyl ether (1.times.500 mL), dried over Na.sub.2SO.sub.4,
filtered through a plug of silica gel, and concentrated in vacuo to
afford a brown solid. The crude material was diluted in a solution
of isopropyl alcohol (520 mL), water (260 mL), and ethylene glycol
(340 g) and then cooled to 0.degree. C. Potassium hydroxide (130 g,
2.32 mol) was added and the solution was heated to 100.degree. C.
for 4 h. After the completion of the reaction, the mixture was
cooled back to 0.degree. C., acidified to pH 3 with aqueous 3 M
HCl, and extracted with EtOAc (2.times.300 mL). The organic layers
were dried over Na.sub.2SO.sub.4, filtered through a plug of silica
gel, and concentrated in vacuo to obtain
2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)propionic
acid.
[0140] c) Oxalyl chloride (2.10 mL, 24.0 mmol) and
dimethylformamide(0.10 mL) were sequentially added to a stirred
solution of
2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)propionic
acid (7.21 g, 18.5 mmol) in dichloromethane (100 mL) at room
temperature. After 1.5 h, the reaction mixture was concentrated and
re-dissolved in dichloromethane (100 mL). Triethylamine (7.77 mL,
55.2 mmol) and (S)-(+)-phenylglycinol (2.54 g, 18.5 mmol) were
added and the mixture was allowed to stir at room temperature until
the reaction was complete (1 h). The mixture was concentrated,
diluted in EtOAc (300 mL), and washed with aqueous 1 M HCl
(1.times.200 mL). The organic layer was dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. Purification
of the crude material by flash chromatography (1-8% THF in DCM)
separated the diastereomers of
2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)-N-(2-hydroxy-1-p-
henyl-ethyl)propanamide. The second-eluting diastereomer was taken
through subsequent transformations.
[0141] d) To a solution of
2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)-N-(2-hydroxy-1-p-
henyl-ethyl)propanamide (3.95 g, 7.76 mmol) and acetic acid (4 mL)
in methanol (80 mL) was added iron powder (4.0 g, 71.7 mmol) and
the reaction mixture was heated to 70.degree. C. for 2 h. After
cooling to room temperature, the mixture was diluted with EtOAc
(150 mL) and washed with 1 M HCl (1.times.100 mL) and water
(1.times.100 mL). The organic layer was dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. Purification
of the crude material by flash chromatography (0-40% EtOAc in
hexanes) gave
(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one.
Example 2: Alternate synthesis of
(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one
##STR00018##
[0143] A solution of
2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)propionic
acid (37.3 g, 95.6 mmol) and (S)-1-(4-chlorophenyl)ethylamine in
isopropyl alcohol (150 mL) and water (50 mL) was heated at
100.degree. C. until all solids were dissolved. The solution was
then allowed to gradually cool to room temperature and left to sit
undisturbed overnight. The salt that came out of solution was
filtered and washed with 2:1 IPA-H.sub.2O (180 mL) to give a pure
crystalline material (17.0 g, 33%, er>100:1 as the free from). A
solution of crystalline (S)-1-(4-chlorophenyl)ethylamine salt of
2-(5-chloro-2-nitro-phenyl)-2-(2,2-dimethylchroman-6-yl)propionic
acid (546 mg, 1.0 mmol), iron powder (224 mg, 4.0 mmol), and acetic
acid (480 mg, 8.0 mmol) in methanol (5.0 mL) was heated to
70.degree. C. for 1 h. After cooling to room temperature, the
mixture was diluted with EtOAc (50 mL) and washed with 1 M HCl
(1.times.50 mL) and water (1.times.50 mL). The organic layer was
dried over Na.sub.2SO.sub.4, filtered, and concentrated in vacuo.
Purification by flash chromatography (0-40% EtOAc in hexanes)
provided
(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one.
Example 3: Synthesis of
3-[(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-2-oxo-indolin-1-yl-
]benzoic acid
##STR00019##
[0145] To a solution of
(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one
(45 mg, 0.13 mmol), tert-butyl-3-iodobenzoate (80 mg, 0.27 mmol),
trans-N,N'-dimethylcyclohexane-1,2-diamine (10 mg, 0.07 mmol), and
potassium carbonate (70 mg, 0.51 mmol) in dioxane (2.5 mL) was
added copper iodide (10 mg, 0.053 mmol). The mixture was purged
with nitrogen and heated to 100.degree. C. After 1 h, the mixture
was cooled to room temperature and diluted with EtOAc (20 mL). The
organic layer was washed with 1 M HCl (1.times.20 mL), water
(1.times.20 mL), dried over Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. The crude material was diluted in
dichloromethane (1 mL) and trifluoroacetic acid (2 mL) and stirred
at room temperature for 5 h. The mixture was concentrated and
purified by reverse-phase HPLC (C18 column, acetonitrile-H.sub.2O
with 0.10% TFA as eluent) to afford the titled compound. .sup.1H
NMR (400 MHz, Chloroform-d) .delta. 8.22-8.09 (m, 2H), 7.69 (dddd,
J=8.0, 1.8, 1.3, 0.4 Hz, 1H), 7.66-7.58 (m, 1H), 7.26-7.17 (m, 2H),
7.06 (d, J=2.3 Hz, 1H), 6.99 (ddd, J=8.5, 2.5, 0.6 Hz, 1H),
6.88-6.81 (m, 1H), 6.73 (d, J=8.6 Hz, 1H), 2.76 (t, J=6.7 Hz, 2H),
1.87 (s, 3H), 1.78 (t, J=6.7 Hz, 2H), 1.32 (d, J=1.5 Hz, 6H); MS:
(ES) m/z calculated for C.sub.27H.sub.25ClNO.sub.4 [M+H].sup.+
462.1, found 462.5.
Example 4: Synthesis of
4-[[(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-2-oxo-indolin-1-y-
l]methyl]benzoic acid
##STR00020##
[0147] To a cooled (0.degree. C.) solution of
(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one
(45 mg, 0.13 mmol) in anhydrous dimethylformamide (0.80 mL) under
nitrogen was added sodium hydride (20 mg, 60% suspension in mineral
oil, 0.50 mmol). After stirring at 0.degree. C. for 10 min, the
solution was allowed to warm to room temperature and methyl
4-(chloromethyl)benzoate (25 mg, 0.14 mmol) was added. The mixture
was left to stir for 30 min at room temperature before the reaction
was carefully quenched by the addition of 1 M HCl (25 mL) and
extracted with EtOAc (50 mL). The organic layer was washed with
water (1.times.20 mL), dried over Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. The crude material was re-dissolved in a
mixture of ethanol (2 mL) and water (1 mL). Lithium hydroxide
monohydrate (100 mg, 2.4 mmol) was added and the mixture was
stirred at 50.degree. C. for 30 min. After cooling to room
temperature, the reaction was quenched by the addition of 1 M HCl
(25 mL) and extracted with EtOAc (50 mL). The organic layer was
washed with water (1.times.20 mL), dried over Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. Purification of the crude
material by reverse-phase HPLC (C18 column, acetonitrile-H.sub.2O
with 0.1% TFA as eluent) gave the titled compound. .sup.1H NMR (400
MHz, Chloroform-d) .delta. 8.05 (d, J=8.2 Hz, 2H), 7.34 (d, J=8.1
Hz, 2H), 7.19-7.12 (m, 2H), 6.99 (d, J=2.4 Hz, 1H), 6.91 (dd,
J=8.6, 2.5 Hz, 1H), 6.72 (d, J=8.5 Hz, 1H), 6.67-6.60 (m, 1H),
5.09-4.89 (m, 2H), 2.74 (t, J=6.8 Hz, 2H), 1.81 (s, 3H), 1.78 (t,
J=6.7 Hz, 2H), 1.32 (s, 6H); MS: (ES) m/z calculated for
C.sub.28H.sub.27ClNO.sub.4 [M+H].sup.+ 476.1, found 476.2.
Example 5: Synthesis of
3-[(3R)-3-(4-tert-butylphenyl)-5-chloro-3-methyl-2-oxo-indolin-1-yl]benzo-
ic acid
##STR00021##
[0149] a) To a cooled (0.degree. C.) solution of sodium hydride
(1.30 g, 39.9 mmol) in anhydrous dimethylformamide (5 mL) under
nitrogen atmosphere was slowly added methyl
p-tert-butylphenylacetate (1.70 g, 8.22 mmol) in dimethylformamide
(5 mL) and the mixture was allowed to stir at 0.degree. C. for 30
min. Next, 4-chloro-2-fluoronitrobenzene (1.60 g, 9.05 mmol) in
dimethylformamide (3 mL) was added dropwise over 10 min. After 1.5
h of stirring at 0.degree. C., methyl iodide (0.51 mL, 16.4 mmol)
was added and the mixture was allowed to warm to room temperature
and stir for an additional 3 h. The reaction was quenched by the
addition of saturated aqueous NH.sub.4Cl (50 mL). The mixture was
extracted with EtOAc (2.times.50 mL) and the organic layers were
dried over Na.sub.2SO.sub.4, filtered, and concentrated in vacuo.
Purification by flash chromatography (0-20% EtOAc in hexanes)
provided methyl
2-(4-tert-butylphenyl)-2-(3-chlorophenyl)propionate. The compound
was dissolved in isopropyl alcohol (40 mL) and water (20 mL) with
potassium hydroxide (2.3 g, 41.1 mmol) and the mixture was heated
at 100.degree. C. for 2 h. The mixture was cooled to room
temperature, acidified to pH 3 with 1 M HCl, and extracted with
EtOAc (2.times.50 mL). The combined organic layers were dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. Purification
of the crude material by flash chromatography (0-30% EtOAc in
hexanes) provided
2-(4-tert-butylphenyl)-2-(3-chlorophenyl)propionic acid.
[0150] b) Oxalyl chloride (0.57 mL, 6.6 mmol) and dimethylformamide
(4 drops) were sequentially added to a stirred solution of
2-(4-tert-butylphenyl)-2-(3-chlorophenyl)propionic acid (2.11 g,
5.5 mmol) in dichloromethane (35 mL) at room temperature. After 2
h, the reaction mixture was concentrated and re-dissolved in
dichloromethane (30 mL). Triethylamine (2.3 mL, 16.5 mmol) and
(S)-(+)-phenylglycinol (750 mg, 5.5 mmol) were added and the
mixture was allowed to stir at room temperature for 1 h. The
mixture was concentrated, diluted in EtOAc (100 mL) and washed with
aqueous 1 M HCl (1.times.50 mL). The organic layer was dried over
Na.sub.2SO.sub.4, filtered, and concentrated in vacuo. Purification
of the crude material by flash chromatography (1-15% EtOAc in DCM)
separated the diastereomers of
(2R)-2-(4-tert-butylphenyl)-2-(3-chlorophenyl)-N-[(1R)-2-hydroxy-1-phenyl-
-ethyl)]propanamide. The second-eluting diastereomer was taken
through subsequent transformations.
[0151] c) To a solution of
(2R)-2-(4-tert-butylphenyl)-2-(3-chlorophenyl)-N-[(1R)-2-hydroxy-1-phenyl-
-ethyl)]propanamide (250 mg, 0.52 mmol) and acetic acid (0.22 mL)
in methanol (2.6 mL) was added iron powder (87 mg, 1.56 mmol) and
the reaction mixture was heated to 70.degree. C. for 2 h. After
cooling to room temperature, the mixture was diluted with EtOAc (20
mL) and washed with 1 M HCl (1.times.10 mL) and water (1.times.10
mL). The organic layer was dried over Na.sub.2SO.sub.4, filtered,
and concentrated in vacuo. Purification of the crude material by
flash chromatography (0-40% EtOAc in hexanes) gave
(3R)-3-(4-tert-butylphenyl)-5-chloro-3-methyl-indolin-2-one.
[0152] d) To a solution of
(3R)-3-(4-tert-butylphenyl)-5-chloro-3-methyl-indolin-2-one (34 mg,
0.11 mmol), tert-butyl-3-iodobenzoate (65 mg, 0.22 mmol),
trans-N,N'-dimethylcyclohexane-1,2-diamine (10 mg, 0.07 mmol), and
potassium carbonate (70 mg, 0.51 mmol) in dioxane (2.5 mL) was
added copper iodide (10 mg, 0.053 mmol). The mixture was purged
with nitrogen and heated to 100.degree. C. After 1 h, the mixture
was cooled to room temperature and diluted with EtOAc (20 mL). The
organic layer was washed with 1 M HCl (1.times.20 mL), water
(1.times.20 mL), dried over Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. The crude material was diluted in
dichloromethane (1 mL) and trifluoroacetic acid (2 mL) and stirred
at room temperature for 5 h. The mixture was concentrated and
purified by reverse-phase HPLC (C18 column, acetonitrile-H.sub.2O
with 0.1% TFA as eluent) to afford the titled compound. .sup.1H NMR
(400 MHz, Chloroform-d) .delta. 8.18-8.10 (m, 2H), 7.72-7.59 (m,
2H), 7.40-7.35 (m, 2H), 7.29 (d, J=2.1 Hz, 1H), 7.28-7.26 (m, 1H),
7.24 (d, J=7.7 Hz, 2H), 6.87-6.81 (m, 1H), 1.91 (s, 3H), 1.30 (s,
9H); MS: (ES) m/z calculated for C.sub.26H.sub.25ClNO.sub.3
[M+H].sup.+ 434.1, found 434.2.
Example 6: Synthesis of
5-[(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-2-oxo-indolin-1-yl-
]-2-methoxy-benzoic acid
##STR00022##
[0154] To a solution of
(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methylindolin-2-one
(60 mg, 0.18 mmol), methyl 5-iodo-2-methoxybenzoate (80 mg, 0.27
mmol), trans-N,N'-dimethylcyclohexane-1,2-diamine (10 mg, 0.07
mmol), and potassium carbonate (102 mg, 0.73 mmol)) in dioxane (3.0
mL) was added copper iodide (20 mg, 0.10 mmol). The mixture was
purged with nitrogen and heated to 100.degree. C. After 1 h, the
mixture was cooled to room temperature and diluted with EtOAc (20
mL). The organic layer was washed with 1 M HCl (1.times.20 mL),
water (1.times.20 mL), dried over Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. The crude material was re-dissolved in a
mixture of ethanol (2 mL) and water (1 mL). Lithium hydroxide
monohydrate (100 mg, 2.4 mmol) was added and the mixture was
stirred at 50.degree. C. for 30 min. After cooling to room
temperature, the reaction was quenched by the addition of 1 M HCl
(25 mL) and extracted with EtOAc (50 mL). The organic layer was
washed with water (1.times.20 mL), dried over Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. Purification of the crude
material by reverse-phase HPLC (C18 column, acetonitrile-H.sub.2O
with 0.1% TFA as eluent) gave the titled compound. .sup.1H NMR (400
MHz, Chloroform-d) .delta. 8.21 (d, J=2.7 Hz, 1H), 7.66 (dd, J=8.9,
2.7 Hz, 1H), 7.29-7.16 (m, 3H), 7.01 (d, J=2.4 Hz, 1H), 6.96 (dd,
J=8.6, 2.5 Hz, 1H), 6.78 (dd, J=8.1, 0.7 Hz, 1H), 6.73 (d, J=8.6
Hz, 1H), 4.13 (s, 3H), 2.75 (t, J=6.7 Hz, 2H), 1.85 (s, 3H), 1.78
(t, J=6.7 Hz, 2H), 1.32 (s, 6H); MS: (ES) m/z calculated for
C.sub.28H.sub.27ClNO.sub.5 [M+H].sup.+ 492.2, found 492.3.
Example 7: Synthesis of
5-[(3R)-3-(2,2-dimethylchroman-6-yl)-3,5-dimethyl-2-oxo-indolin-1-yl]-2-m-
ethyl-benzoic acid
##STR00023##
[0156] To a solution of
(3R)-3-(2,2-dimethylchroman-6-yl)-3,5-dimethylindolin-2-one (32 mg,
0.10 mmol), methyl 5-iodo-2-methylbenzoate (36 mg, 0.13 mmol),
trans-N,N'-dimethylcyclohexane-1,2-diamine (10 mg, 0.07 mmol), and
potassium carbonate(28 mg, 0.20 mmol)) in dioxane (3.0 mL) was
added copper iodide (6.0 mg, 0.03 mmol). The mixture was purged
with nitrogen and heated to 100.degree. C. After 1 h, the mixture
was cooled to room temperature and diluted with EtOAc (20 mL). The
organic layer was washed with 1 M HCl (1.times.20 mL), water
(1.times.20 mL), dried over Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. The crude material was re-dissolved in a
mixture of ethanol (2 mL) and water (1 mL). Lithium hydroxide
monohydrate (100 mg, 2.4 mmol) was added and the mixture was
stirred at 50.degree. C. for 30 min. After cooling to room
temperature, the reaction was quenched by the addition of 1 M HCl
(25 mL) and extracted with EtOAc (50 mL). The organic layer was
washed with water (1.times.20 mL), dried over Na.sub.2SO.sub.4,
filtered, and concentrated in vacuo. Purification of the crude
material by reverse-phase HPLC (C18 column, acetonitrile-H.sub.2O
with 0.1% TFA as eluent) gave the titled compound. .sup.1H NMR (400
MHz, Chloroform-d) .delta. 8.19 (t, J=1.9 Hz, 1H), 8.13-8.06 (m,
1H), 7.76-7.69 (m, 1H), 7.61 (t, J=7.9 Hz, 1H), 7.14-7.03 (m, 3H),
7.03-6.97 (m, 1H), 6.86-6.77 (m, 1H), 6.71 (d, J=8.6 Hz, 1H), 2.75
(t, J=6.8 Hz, 2H), 2.36 (d, J=0.9 Hz, 3H), 1.85 (s, 3H), 1.78 (t,
J=6.7 Hz, 2H), 1.31 (d, J=1.8 Hz, 6H); MS: (ES) m/z calculated for
C.sub.29H.sub.30NO.sub.4 [M+H].sup.+ 456.6, found 456.0.
Example 8: Synthesis of
2-[4-[(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-2-oxo-indolin-1-
-yl]phenyl]acetic acid
##STR00024##
[0158] To a solution of
(3R)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-indolin-2-one
(60 mg, 0.18 mmol), 4-iodophenylacetic acid (92 mg, 0.35 mmol),
trans-N,N'-dimethylcyclohexane-1,2-diamine (10 mg, 0.07 mmol), and
potassium carbonate (97 mg, 0.70 mmol) in dimethylformamide (3.0
mL) was added copper iodide (20 mg, 0.10 mmol). The mixture was
purged with nitrogen and heated to 110.degree. C. After 1 h, the
mixture was cooled to room temperature and diluted with EtOAc (20
mL). The organic layer was washed with 1 M HCl (1.times.20 mL),
water (1.times.20 mL), dried over Na.sub.2SO.sub.4, filtered, and
concentrated in vacuo. Purification of the crude material by
reverse-phase HPLC (C18 column, acetonitrile-H.sub.2O with 0.1% TFA
as eluent) gave the titled compound. .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 7.49-7.41 (m, 2H), 7.41-7.34 (m, 2H), 7.20
(dq, J=4.3, 2.1 Hz, 2H), 7.04 (d, J=2.3 Hz, 1H), 6.96 (ddd, J=8.6,
1.9, 1.2 Hz, 1H), 6.85 (d, J=8.9 Hz, 1H), 6.71 (d, J=8.5 Hz, 1H),
3.71 (s, 2H), 2.74 (t, J=6.7 Hz, 2H), 1.84 (s, 3H), 1.78 (t, J=6.7
Hz, 2H), 1.31 (d, J=1.8 Hz, 6H); MS: (ES) m/z calculated for
C.sub.29H.sub.27ClNO.sub.4 [M+H].sup.+ 476.2, found 476.2.
Example 9: Synthesis of
(S)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-1-(4-((5-oxo-4,5-dihyd-
ro-1H-tetrazol-1-yl)methyl)phenyl)indolin-2-one
##STR00025##
[0160] Step a: To a solution of
(S)-2-(4-(5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-2-oxoindolin-1-y-
l)phenyl)acetic acid (190.4 mg, 0.40 mmol) in dichloromethane (1.6
mL) at 0.degree. C. under nitrogen, a solution of oxalyl chloride
(52 .mu.L, 0.60 mmol) in dichloromethane (0.3 mL) was added
dropwise. The reaction mixture was stirred at 0.degree. C. for 5
min and then warmed to room temperature and stirred for 4 hours.
All solvents were removed under vacuum and dichloromethane (2 mL)
was added to the residue. The mixture was concentrated under vacuum
and this process was repeated another time to give the acid
chloride product which was used in the next step directly.
[0161] Step b: To the acid chloride prepared in the previous step
was added azidotrimethylsilane (0.32 mL, 2.4 mmol) at room
temperature (gas evolution!). The mixture was heated to 100.degree.
C. under nitrogen and stirred for 37 h. The reaction mixture was
cooled to room temperature and concentrated under vacuum. Water and
dichloromethane were added to the residue. The organic layer was
separated, dried, and concentrated. The crude product was purified
by silica gel chromatography (50% ethyl acetate/hexane) to give the
desired product. .sup.1H NMR (400 MHz, chloroform-d) .delta.
7.58-7.51 (m, 2H), 7.47-7.41 (m, 2H), 7.22-7.16 (m, 2H), 7.03 (d,
J=2.4 Hz, 1H), 6.96 (dd, J=8.6, 2.5 Hz, 1H), 6.85-6.80 (m, 1H),
6.70 (d, J=8.6 Hz, 1H), 5.16 (s, 2H), 2.73 (t, J=6.7 Hz, 2H), 1.83
(s, 3H), 1.77 (t, J=6.7 Hz, 2H), 1.30 (d, J=2.0 Hz, 6H). MS: (ES)
m/z calculated for C.sub.28H.sub.26ClNO.sub.3[M+H].sup.+ 516.2,
found 516.5.
Example 10: Synthesis of
(S)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-1-(4-((4-methyl-5-oxo--
4,5-dihydro-1H-tetrazol-1-yl)methyl)phenyl)indolin-2-one
##STR00026##
[0163] To a mixture of
(S)-5-chloro-3-(2,2-dimethylchroman-6-yl)-3-methyl-1-(4-((5-oxo-4,5-dihyd-
ro-1H-tetrazol-1-yl)methyl)phenyl)indolin-2-one (204 mg, 0.395
mmol) and iodomethane (49 .mu.L, 0.791 mmol) in DMF (1.0 mL) at
0.degree. C. was added K.sub.2CO.sub.3 (138 mg, 1.0 mmol). The
reaction mixture was stirred at 0.degree. C. for 20 min and then
room temperature for 14 h. The mixture was poured into water (10
mL) and dichloromethane (3 mL). The organic layer was separated and
the aqueous layer was extracted with dichloromethane. The combined
organic layers were dried and concentrated. The crude product was
purified by silica gel chromatography (50% ethyl acetate/hexane) to
give the desired product. .sup.1H NMR (400 MHz, chloroform-d)
.delta. 7.58-7.52 (m, 2H), 7.44-7.39 (m, 2H), 7.21-7.16 (m, 2H),
7.03 (d, J=2.4 Hz, 1H), 6.98-6.93 (m, 1H), 6.85-6.80 (m, 1H), 6.70
(d, J=8.6 Hz, 1H), 5.13 (s, 2H), 3.61 (s, 3H), 2.73 (t, J=6.7 Hz,
2H), 1.82 (s, 3H), 1.76 (t, J=6.7 Hz, 2H), 1.30 (d, J=1.8 Hz, 6H).
MS: (ES) m/z calculated for
C.sub.29H.sub.28ClN.sub.5O.sub.3[M+H].sup.+ 530.2, found 530.5.
[0164] Compounds prepared by methods analogous to the methods
described above, and evaluated using the serum chemotaxis assay
below are provided in the following table. A.sub.2 was calculated
as described and activity is presented in Table 1 as: +, 20000
nM.gtoreq.A.sub.2.gtoreq.500 nM; ++, 500 nM>A.sub.2.gtoreq.100
nM; +++, 100 nM>A.sub.2.
TABLE-US-00001 TABLE 1 MS: (ES) Chemotaxis .sup.1H NMR m/z A.sub.2
##STR00027## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.28-8.19
(m, 2H), 7.59 (dt, J = 8.4, 0.5 Hz, 2H), 7.34 (d, J = 9.1 Hz, 2H),
7.31-7.27 (m, 2H), 7.17 (d, J = 7.6 Hz, 1H), 7.03-6.94 (m, 1H),
6.85-6.78 (m, 1H), 2.36 (s, 3H), 1.89 (s, 3H), 1.29 (s, 9H). 414.3
[M + H].sup.+ ++ ##STR00028## .sup.1H NMR (400 MHz,
Methanol-d.sub.4) .delta. 8.28-8.17 (m, 2H), 7.66-7.56 (m, 2H),
7.41 (d, J = 8.5 Hz, 2H), 7.31 (dd, J = 8.4, 2.2 Hz, 1H), 7.27 (s,
2H), 7.25 (d, J = 0.6 Hz, 1H), 6.96 (d, J = 8.4 Hz, 1H), 1.88 (s,
3H), 1.30 (s, 9H). 434.1 [M + H].sup.+ +++ ##STR00029## .sup.1H NMR
(400 MHz, Chloroform-d) .delta. 8.21-8.12 (m, 1H), 7.72 (dtd, J =
9.3, 7.7, 1.6 Hz, 1H), 7.54 (td, J = 7.6, 1.1 Hz, 1H), 7.44-7.36
(m, 1H), 7.36-7.32 (m, 2H), 7.32- 7.27 (m, 2H), 7.24-7.14 (m, 2H),
6.53 (dd, J = 17.9, 8.7 Hz, 1H), 1.76 (s, 3H), 1.28 (d, J = 12.5
Hz, 9H). 434.1 [M + H].sup.+ ++ ##STR00030## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.19-8.07 (m, 2H), 7.78-7.58 (m, 2H),
7.21-7.16 (m, 1H), 7.13 (dd, J = 8.0, 1.9 Hz, 1H), 7.05 (d, J = 2.4
Hz, 1H), 7.00 (dd, J = 8.6, 2.5 Hz, 1H), 6.88 (d, J = 1.8 Hz, 1H),
6.72 (d, J = 8.6 Hz, 1H), 2.74 (t, J = 6.7 Hz, 2H), 1.85 (s, 3H),
1.78 (t, J = 6.7 Hz, 2H), 1.33-1.30 (m, 6H). 461.9 [M + H].sup.+ ++
##STR00031## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.14 (dt,
J = 4.1, 1.9 Hz, 2H), 7.71-7.60 (m, 2H), 7.30-7.26 (m, 1H), 7.25
(q, J = 1.8 Hz, 2H), 7.06-6.96 (m, 2H), 6.86 (d, J = 8.2 Hz, 1H),
1.89 (s, 3H), 1.36 (d, J = 1.0 Hz, 9H). 474.1 [M + Na].sup.+ +++
##STR00032## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.20-8.06
(m, 2H), 7.71 (d, J = 7.9 Hz, 1H), 7.66-7.57 (m, 1H), 7.35 (dd, J =
8.6, 1.2 Hz, 2H), 7.32- 7.28 (m, 2H), 7.11-7.03 (m, 2H), 6.83 (d, J
= 8.0 Hz, 1H), 2.36 (s, 4H), 1.90 (d, J = 1.2 Hz, 3H), 1.30 (d, J =
1.2 Hz, 9H). 414.2 [M + H].sup.+ ++ ##STR00033## .sup.1H NMR (400
MHz, Chloroform-d) .delta. 8.22 (d, J = 8.5 Hz, 2H), 7.60 (d, J =
8.6 Hz, 2H), 7.35 (d, J = 8.6 Hz, 2H), 7.31-7.27 (m, 2H), 7.08 (d,
J = 10.0 Hz, 2H), 6.92 (d, J = 7.9 Hz, 1H), 2.36 (s, 3H), 1.89 (s,
3H), 1.29 (d, J = 0.4 Hz, 9H). 414.2 [M + H].sup.+ ++ ##STR00034##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.33-8.24 (m, 2H),
7.65-7.60 (m, 2H), 7.58 (d, J = 8.4 Hz, 1H), 7.35-7.29 (m, 1H),
7.25-7.19 (m, 1H), 7.10 (dd, J = 2.3, 0.8 Hz, 1H), 6.98-6.88 (m,
2H), 1.90 (s, 3H), 1.84 (s, 3H), 1.31 (d, J = 0.4 Hz, 9H). 448.1 [M
+ H].sup.+ ++ ##STR00035## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.20-8.14 (m, 2H), 7.77-7.71 (m, 1H), 7.71-7.65 (m, 1H),
7.58 (d, J = 8.3 Hz, 1H), 7.35-7.31 (m, 1H), 7.20 (dd, J = 8.4, 2.2
Hz, 1H), 7.13-7.09 (m, 1H), 6.94 (d, J = 2.1 Hz, 1H), 6.84 (d, J =
8.4 Hz, 1H), 1.90 (s, 3H), 1.86 (s, 3H), 1.31 (s, 9H). 448.1 [M +
H].sup.+ +++ ##STR00036## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.18-8.09 (m, 2H), 7.71-7.66 (m, 1H), 7.63 (dd, J = 8.2,
7.4 Hz, 1H), 7.27-7.24 (m, 2H), 7.23 (dq, J = 1.5, 0.7 Hz, 2H),
6.88-6.80 (m, 3H), 4.52 (p, J = 6.1 Hz, 1H), 1.88 (s, 3H), 1.32 (d,
J = 6.0 Hz, 6H). 436.1 [M + H].sup.+ +++ ##STR00037## .sup.1H NMR
(400 MHz, Chloroform-d) .delta. 8.15 (dd, J = 9.0, 1.7 Hz, 1H),
8.02 (d, J = 22.0 Hz, 1H), 7.62- 7.50 (m, 2H), 7.37 (d, J = 8.1 Hz,
3H), 7.24 (dd, J = 1.6, 0.8 Hz, 2H), 7.14 (d, J = 2.0 Hz, 1H), 1.90
(s, 3H), 1.30 (d, J = 0.5 Hz, 9H). 468.2 [M + H].sup.+ +
##STR00038## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.21-8.12
(m, 1H), 7.72 (dtd, J = 9.3, 7.7, 1.6 Hz, 1H), 7.54 (td, J = 7.6,
1.1 Hz, 1H), 7.44-7.36 (m, 1H), 7.36-7.32 (m, 2H), 7.32- 7.27 (m,
2H), 7.24-7.14 (m, 2H), 6.53 (dd, J = 17.9, 8.7 Hz, 1H), 1.76 (s,
3H), 1.28 (d, J = 12.5 Hz, 9H). 434.2 [M + H].sup.+ ++ ##STR00039##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.17 (s, 1H), 8.11 (d,
J = 7.7 Hz, 1H), 7.74-7.67 (m, 1H), 7.62 (t, J = 7.8 Hz, 1H),
7.25-7.23 (m, 1H), 7.09-7.05 (m, 3H), 7.01 (dd, J = 13.9, 2.1 Hz,
1H), 6.83 (d, J = 8.4 Hz, 1H), 2.37 (d, J = 0.9 Hz, 3H), 1.87 (d, J
= 0.9 Hz, 3H), 1.35 (t, J = 1.0 Hz, 9H). 432.2 [M + H].sup.+ +++
##STR00040## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.20-8.08
(m, 2H), 7.68-7.63 (m, 2H), 7.26-7.23 (m, 1H), 7.19 (d, J = 7.9 Hz,
1H), 7.16-7.11 (m, 1H), 7.07-6.95 (m, 2H), 6.88 (d, J = 1.8 Hz,
1H), 1.87 (d, J = 0.7 Hz, 3H), 1.35 (d, J = 0.9 Hz, 9H). 452.2 [M +
H].sup.+ + ##STR00041## .sup.1H NMR (400 MHz, Chloroform-d) .delta.
8.29-8.22 (m, 2H), 7.61-7.54 (m, 2H), 7.25-7.21 (m, 4H), 7.00- 6.93
(m, 1H), 6.87-6.80 (m, 2H), 4.61-4.38 (m, 1H), 1.88 (s, 3H), 1.32
(dd, J = 6.1, 1.2 Hz, 6H). 436.2 [M + H].sup.+ +++ ##STR00042##
452.2 [M + H].sup.+ ++ ##STR00043## 452.1 [M + H].sup.+ +++
##STR00044## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.18-8.12
(m, 2H), 7.72-7.68 (m, 1H), 7.64 (t, J = 7.8 Hz, 1H), 7.25-7.21 (m,
1H), 7.16 (d, J = 7.6 Hz, 1H), 7.10-7.05 (m, 1H), 7.04-6.96 (m,
2H), 6.71 (t, J = 1.2 Hz, 1H), 2.36 (d, J = 0.9 Hz, 3H), 1.87 (s,
3H), 1.35 (t, J = 0.7 Hz, 9H). 432.3 [M + H].sup.+ ++ ##STR00045##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.17 (t, J = 1.9 Hz,
1H), 8.12 (dt, J = 7.5, 1.4 Hz, 1H), 7.71-7.67 (m, 1H), 7.63 (t, J
= 7.8 Hz, 1H), 7.35-7.32 (m, 2H), 7.32-7.28 (m, 2H), 7.17 (d, J =
7.6 Hz, 1H), 6.99-6.92 (m, 1H), 6.72-6.69 (m, 1H), 2.35 (d, J = 0.7
Hz, 3H), 1.89 (s, 3H), 1.29 (d, J = 0.5 Hz, 9H). 414.2 [M +
H].sup.+ ++ ##STR00046## 452.1 [M + H].sup.+ ++ ##STR00047##
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.86 (dd, J = 5.3, 0.8
Hz, 1H), 8.36- 8.08 (m, 1H), 7.84 (dd, J = 5.3, 2.1 Hz, 1H), 7.47
(d, J = 2.2 Hz, 1H), 7.38 (d, J = 2.6 Hz, 1H), 7.36 (d, J = 2.4 Hz,
2H), 7.24 (d, J = 8.5 Hz, 2H), 7.16 (d, J = 8.5 Hz, 1H), 1.84 (s,
3H), 1.23 (s, 9H). 435.0 [M + H].sup.+ ++ ##STR00048## .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 9.11 (d, J = 1.9 Hz, 1H), 8.97 (d,
J = 2.4 Hz, 1H), 8.40 (ddd, J = 2.5, 1.9, 0.6 Hz, 1H), 7.45 (d, J =
2.2 Hz, 1H), 7.37 (d, J = 8.5 Hz, 2H), 7.32 (ddd, J = 8.5, 2.2, 0.6
Hz, 1H), 7.30- 7.26 (m, 2H), 6.95 (d, J = 8.5 Hz, 1H), 1.84 (s,
3H), 1.24 (d, J = 0.6 Hz, 9H). 435.0 [M + H].sup.+ ++ ##STR00049##
.sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.81-8.77 (m, 1H), 8.20
(dd, J = 1.5, 0.8 Hz, 1H), 7.87-7.78 (m, 1H), 7.71 (d, J = 8.6 Hz,
1H), 7.45 (d, J = 2.3 Hz, 1H), 7.41-7.34 (m, 3H), 7.27-7.19 (m,
2H), 1.84 (s, 3H), 1.23 (s, 9H). 435.0 [M + H].sup.+ +++
##STR00050## .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.19-8.13
(m, 1H), 8.10-8.00 (m, 2H), 7.97 (dt, J = 8.6, 0.5 Hz, 1H),
7.48-7.44 (m, 1H), 7.41 (ddd, J = 8.6, 2.3, 0.6 Hz, 1H), 7.39- 7.33
(m, 2H), 7.24-7.19 (m, 2H), 1.84 (s, 3H), 1.23 (d, J = 0.6 Hz, 9H).
456.9 [M + Na].sup.+ ++ ##STR00051## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.14 (d, J = 5.4 Hz, 2H), 7.66 (d J = 7.1 Hz,
2H), 7.35-7.31 (m, 2H), 7.31 (d, J = 1.8 Hz, 2H), 7.29- 7.27 (m,
1H), 7.23-7.21 (m, 1H), 6.86 (d, J = 8.4 Hz, 1H), 1.89 (s, 3H).
434.1 [M + Na].sup.+ + ##STR00052## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 7.84 (d, J = 64.1 Hz, 2H), 7.24- 7.13 (m,
2H), 7.08 (d, J = 1.8 Hz, 1H), 7.03 (d, J = 7.8 Hz, 1H), 6.99- 6.87
(m, 2H), 6.78 (t, J = 8.6 Hz, 1H), 6.63 (s, 1H), 3.74 (s, 3H), 1.63
(s, 3H). 426.2 [M + H].sup.+ + ##STR00053## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.17-8.11 (m, 2H), 7.69 (dt, J = 7.9, 1.7 Hz,
1H), 7.64 (t, J = 8.0 Hz, 1H), 7.52-7.41 (m, 2H), 7.37- 7.28 (m,
2H), 7.25-7.22 (m, 1H), 6.86 (dd, J = 8.1, 0.8 Hz, 1H), 1.92 (s,
3H), 1.57 (s, 6H). 457.9 [M + Na].sup.+ ++ ##STR00054## .sup.1H NMR
(400 MHz, DMSO-d.sub.6) .delta. 8.03 (t, J = 8.3 Hz, 1H), 7.57 (dd,
J = 11.5, 1.9 Hz, 1H), 7.45 (td, J = 4.1, 1.9 Hz, 2H), 7.39-7.29
(m, 3H), 7.28-7.22 (m, 2H), 7.02 (d, J = 8.5 Hz, 1H), 1.82 (s, 3H),
1.24 (d, J = 0.6 Hz, 9H). 452.1 [M + H].sup.+ ++ ##STR00055##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.13 (t, J = 7.1 Hz,
1H), 7.76- 7.58 (m, 1H), 7.47-7.32 (m, 3H), 7.32-7.26 (m, 2H),
7.26-7.19 (m, 2H), 6.63 (dd, J = 8.6, 5.5 Hz, 1H), 1.92 (s, 3H),
1.30 (dd, J = 5.2, 1.0 Hz, 9H). 452. 1 [M + H].sup.+ +++
##STR00056## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.16-8.12
(m, 2H), 7.72-7.61 (m, 2H), 7.39-7.36 (m, 2H), 7.36- 7.32 (m, 2H),
7.24 (t, J = 1.1 Hz, 2H), 6.86 (dd, J = 8.1, 0.9 Hz, 1H), 1.92 (d,
J = 1.0 Hz, 3H), 1.67 (d, J = 21.9 Hz, 6H). 459.9 [M + Na].sup.+ ++
##STR00057## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.14 (d, J
= 4.1 Hz, 2H), 7.74- 7.62 (m, 2H), 7.57 (d, J = 8.5 Hz, 2H), 7.39
(d, J = 8.2 Hz, 2H), 7.26- 7.23 (m, 2H), 6.87 (d, J = 8.1 Hz, 1H),
1.93 (s, 3H), 1.76 (s, 3H). 511.8 [M + Na].sup.+ ++ ##STR00058##
.sup.1H NMR (400 MHz, Methanol-d.sub.4) .delta. 8.17-8.11 (m, 1H),
8.10-8.05 (m, 1H), 7.72 (d, J = 1.3 Hz, 1H), 7.71-7.70 (m, 1H),
7.42 (d, J = 2.0 Hz, 1H), 7.41 (d, J = 2.0 Hz, 1H), 7.27 (d, J =
8.5 Hz, 2H), 7.22 (q, J = 1.0 Hz, 2H), 7.00-6.94 (m, 1H), 1.91 (s,
3H), 1.31 (s, 9H). 484.2 [M + H].sup.+ +++ ##STR00059## .sup.1H NMR
(400 MHz, Chloroform-d) .delta. 8.14 (dt, J = 4.0, 1.8 Hz, 2H),
7.68 (d, J = 2.1 Hz, 1H), 7.67- 7.62 (m, 1H), 7.44 (d, J = 8.1 Hz,
2H), 7.35-7.31 (m, 2H), 7.25 (dt, J = 1.0, 0.5 Hz, 2H), 6.90-6.82
(m, 1H), 1.91 (d, J = 0.8 Hz, 3H), 1.44-1.25 (m, 2H), 1.00 (s, 2H).
486.1 [M + H].sup.+ +++ ##STR00060## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.17-8.11 (m, 2H), 7.70-7.61 (m, 2H),
7.37-7.32 (m, 2H), 7.29 (d, J = 2.2 Hz, 1H), 7.28-7.27 (m, 2H),
7.22 (d, J = 2.1 Hz, 1H), 6.86 (d, J = 8.4 Hz, 1H), 1.90 (s, 3H),
1.79-1.60 (m, 2H), 1.44-1.27 (m, 2H). 443.1 [M + H].sup.+ ++
##STR00061## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.17-8.09
(m, 2H), 7.68 (d, J = 8.2 Hz, 1H), 7.66-7.60 (m, 1H), 7.24 (dtd, J
= 2.6, 1.1, 0.6 Hz, 3H), 7.24-7.20 (m, 3H), 6.85 (d, J = 9.0 Hz,
1H), 1.89 (s, 3H), 1.39 (s, 3H), 0.84 (s, 2H), 0.78-0.64 (m, 2H).
432.2 [M + H].sup.+ +++ ##STR00062## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.24-8.09 (m, 2H), 7.71 (ddd, J = 8.0, 2.1,
1.4 Hz, 1H), 7.65 (t, J = 8.0 Hz, 1H), 7.26 (dd, J = 2.6, 0.4 Hz,
1H), 7.25-7.21 (m, 1H), 7.09- 7.06 (m, 1H), 7.05 (dt, J = 8.6, 1.6
Hz, 1H), 6.84 (dt, J = 8.2, 0.5 Hz, 1H), 6.78 (d, J = 8.6 Hz, 1H),
4.49 (p, J = 6.0 Hz, 1H), 2.18 (s, 3H), 1.88 (s, 3H), 1.32 (dd, J =
6.0, 0.5 Hz, 6H). 450.2 [M + H].sup.+ ++ ##STR00063## .sup.1H NMR
(400 MHz, Methanol-d.sub.4) .delta. 7.60-7.56 (m, 2H), 7.56-7.53
(m, 3H), 7.40-7.35 (m, 2H), 7.31- 7.29(m, 1H), 7.28 (q, J = 1.9 Hz,
2H), 6.84 (d, J = 9.1 Hz, 1H), 1.90 (s, 3H), 1.57 (s, 3H), 1.56 (s,
6H). 484.2 [M + H].sup.+ + ##STR00064## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.22-8.11 (m, 2H), 7.73-7.68 (m, 1H), 7.65
(t, J = 7.7 Hz, 1H), 7.14 (d, J = 9.1 Hz, 2H), 7.05 (d, J = 2.4 Hz,
1H), 6.99 (dd, J = 8.5, 2.6 Hz, 1H), 6.94-6.87 (m, 1H), 6.73 (d, J
= 8.6 Hz, 1H), 2.75 (t, J = 6.7 Hz, 2H), 1.88 (s, 3H), 1.78 (t, J =
6.7 Hz, 2H), 1.32 (s, 6H). 512.2 [M + H].sup.+ +++ ##STR00065##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.21-8.16 (m, 1H),
8.15-8.10 (m, 1H), 7.69-7.65 (m, 2H), 7.62- 7.54 (m, 1H), 7.53 (d,
J = 1.6 Hz, 1H), 7.02 (d, J = 2.5 Hz, 1H), 6.99- 6.94 (m, 2H), 6.74
(d, J = 8.6 Hz, 1H), 2.76 (t, J = 6.8 Hz, 2H), 1.89 (s, 3H),
1.82-1.75 (m, 2H), 1.33 (d, J = 1.1 Hz, 6H). 453.2 [M + H].sup.+
+++ ##STR00066## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.16
(dt, J = 10.3, 1.5 Hz, 1H), 8.14-8.06 (m, 1H), 7.76-7.68 (m, 1H),
7.68-7.57 (m, 1H), 7.42- 7.33 (m, 1H), 7.07 (dd, J = 14.5, 2.5 Hz,
1H), 7.02-6.93 (m, 2H), 6.81 (dd, J = 8.3, 7.4 Hz, 1H), 6.75- 6.65
(m, 1H), 2.79-2.67 (m, 2H), 1.86 (s, 1H), 1.85 (s, 2H), 1.78 (td, J
= 6.7, 2.8 Hz, 2H), 1.32 (t, J = 1.7 Hz, 6H), 0.99-0.92 (m, 1H),
0.69-0.56 (m, 1H). 468.2 [M + H].sup.+ +++ ##STR00067## .sup.1H NMR
(400 MHz, Methanol-d.sub.4) .delta. 8.10 (ddd, J = 5.0, 3.5, 1.6
Hz, 1H), 8.06 (d, J = 1.8 Hz, 1H), 7.73- 7.65 (m, 2H), 7.17-7.12
(m, 1H), 7.08 (dd, J = 17.7, 2.0 Hz, 2H), 6.98 (dd, J = 8.6, 2.5
Hz, 1H), 6.81 (d, J = 8.0 Hz, 1H), 6.66 (d, J = 8.6 Hz, 1H), 2.76
(t, J = 6.8 Hz, 2H), 2.64 (q, J = 7.6 Hz, 2H), 1.83 (s, 3H), 1.79
(t, J = 6.8 Hz, 2H), 1.32-1.27 (m, 6H), 1.21 (t, J = 7.6 Hz, 3H).
456.3 [M + H].sup.+ +++ ##STR00068## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.18-8.16 (m, 1H), 8.14 (dt, J = 7.8, 1.4 Hz,
1H), 7.71 (ddd, J = 7.9, 2.1, 1.2 Hz, 1H), 7.67-7.61 (m, 1H),
7.25-7.20 (m, 2H), 6.89- 6.82 (m, 3H), 2.73 (t, J = 6.8 Hz, 2H),
2.13 (t, J = 0.6 Hz, 3H), 1.85 (s, 3H), 1.76 (t, J = 6.7 Hz, 2H),
1.31 (s, 6H). 479.5 [M + H].sup.+ +++ ##STR00069## .sup.1H NMR (400
MHz, Methanol-d.sub.4) .delta. 8.12 (ddd, J = 4.9, 4.1, 1.6 Hz,
1H), 8.05 (q, J = 1.2 Hz, 1H), 7.75- 7.58 (m, 2H), 7.37-7.20 (m,
2H), 6.95-6.86 (m, 2H), 6.85- 6.79 (m, 1H), 2.79 (t, J = 6.8 Hz,
2H), 1.84 (s, 3H), 1.82 (d, J = 6.8 Hz, 2H), 1.33 (s, 6H). 502.1 [M
+ H].sup.+ +++ ##STR00070## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.19-8.12 (m 2H), 7.70 (ddt, J = 8.6, 1.7, 0.9 Hz, 1H),
7.67-7.60 (m, 1H), 7.25-7.21 (m, 2H), 7.13 (t, J = 1.6 Hz, 1H),
7.04 (dd, J = 8.5, 2.1 Hz, 1H), 6.85 (dd, J = 9.1, 1.1 Hz, 1H),
6.73-6.65 (m, 1H), 3.00 (s, 2H), 1.87 (s, 3H), 1.69- 1.24 (m, 6H).
448.5 [M + H].sup.+ +++ ##STR00071## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.07-7.92 (m, 2H), 7.57 (dt, J = 4.9, 1.2 Hz,
2H), 7.43-7.29 (m, 1H), 7.25-7.14 (m, 2H), 7.11- 6.97 (m, 2H),
6.87-6.71 (m, 1H), 2.52 (d, J = 3.9 Hz, 3H), 1.88 (d, J = 3.1 Hz,
3H), 1.38 (dd, J = 1.3, 0.5 Hz, 9H). 448.3 [M +
Na].sup.+ ++ ##STR00072## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.21-8.16 (m, 1H), 8.15 (tt, J = 1.8, 0.8 Hz, 1H),
7.68-7.63 (m, 2H), 7.23 (q, J = 1.3 Hz, 1H), 7.03 (d, J = 2.4 Hz,
1H), 7.01-6.96 (m, 2H), 6.74 (d, J = 8.5 Hz, 1H), 2.75 (t, J = 6.6
Hz, 2H), 1.88 (s, 3H), 1.79 (t, J = 6.8 Hz, 2H), 1.32 (s, 6H).
546.3 [M + H].sup.+ +++ ##STR00073## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.20-8.14 (m, 1H), 8.13 (q, J = 1.5 Hz, 1H),
7.67 (d, J = 1.7 Hz, 1H), 7.66 (d, J = 1.6 Hz, 1H), 7.31 (s, 1H),
7.05-6.98 (m, 2H), 6.97 (s, 1H), 6.74 (d, J = 8.6 Hz, 1H), 2.76 (t,
J = 6.8 Hz, 2H), 1.86 (s, 3H), 1.78 (d, J = 6.8 Hz, 2H), 1.32 (d, J
= 1.5 Hz, 6H). 518.2 [M + Na].sup.+ +++ ##STR00074## .sup.1H NMR
(400 MHz, Chloroform-d) .delta. 8.18-8.10 (m, 2H), 7.72-7.59 (m,
2H), 7.40-7.35 (m, 2H), 7.29 (d, J = 2.1 Hz, 1H), 7.28-7.26 (m,
1H), 7.24 (d, J = 7.7 Hz, 2H), 6.87- 6.81 (m, 1H), 1.91 (s, 3H),
1.30 (s, 9H). 434.2 [M + H].sup.+ +++ ##STR00075## .sup.1H NMR (400
MHz, Chloroform-d) .delta. 8.08 (dd, J = 6.2, 2.7 Hz, 1H), 7.65
(ddd, J = 8.8, 4.0, 2.7 Hz, 1H), 7.44-7.29 (m, 3H), 7.28-7.20 (m,
4H), 6.81 (d, J = 9.0 Hz, 1H), 1.90 (s, 3H), 1.30 (s, 9H). 452.2 [M
+ H].sup.+ +++ ##STR00076## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.13-8.06 (m, 1H), 7.51 (dd, J = 8.1, 2.2 Hz, 1H), 7.42 (d,
J = 8.2 Hz, 1H), 7.39-7.31 (m, 2H), 7.30- 7.17 (m, 4H), 6.85-6.78
(m 1H), 2.69 (s, 3H), 1.89 (s, 3H), 1.30 (s, 9H). 448.2 [M +
H].sup.+ +++ ##STR00077## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.22-8.09 (m, 2H), 7.69 (dddd, J = 8.0, 1.8, 1.3, 0.4 Hz,
1H), 7.66- 7.58 (m, 1H), 7.26-7.17 (m, 2H), 7.06 (d, J = 2.3 Hz,
1H), 6.99 (ddd, J = 8.5, 2.5, 0.6 Hz, 1H), 6.88- 6.81 (m, 1H), 6.73
(d, J = 8.6 Hz, 1H), 2.76 (t, J = 6.7 Hz, 2H), 1.87 (s, 3H), 1.78
(t, J = 6.7 Hz, 2H), 1.32 (d, J = 1.5 Hz, 6H). 462.5 [M + H].sup.+
+++ ##STR00078## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.10
(d, J = 2.3 Hz, 1H), 7.52 (dd, J = 8.1, 2.3 Hz, 1H), 7.42 (dt, J =
8.0, 0.7 Hz, 1H), 7.22-7.19 (m, 2H), 7.06-7.01 (m, 1H), 6.98 (ddd,
J = 8.7, 2.5, 0.7 Hz, 1H), 6.83 (dd, J = 8.9, 0.6 Hz, 1H), 6.73 (d,
J = 8.5 Hz, 1H), 2.80-2.71 (m, 2H), 2.69 (s, 3H), 1.85 (s, 3H),
1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J = 1.6 Hz, 6H). 476.2 [M +
H].sup.+ +++ ##STR00079## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.39-8.13 (m, 2H), 7.65-7.50 (m, 2H), 7.30-7.25 (m, 2H),
7.05 (d, J = 2.3 Hz, 1H), 7.00-6.92 (m, 2H), 6.73 (d, J = 8.6 Hz,
1H), 2.75 (t, J = 6.8 Hz, 2H), 1.86 (s, 3H), 1.78 (t, J = 6.7 Hz,
2H), 1.43-1.08 (m, 6H). 462.2 [M + H].sup.+ +++ ##STR00080##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.08 (dd, J = 6.3, 2.7
Hz, 1H), 7.66 (ddd, J = 8.8, 4.0, 2.7 Hz, 1H), 7.32 (dd, J = 9.9,
8.8 Hz, 1H), 7.27- 7.23 (m, 2H), 7.04 (d, J = 2.5 Hz, 1H), 6.97
(ddd, J = 8.7, 2.5, 0.6 Hz, 1H), 6.87-6.78 (m, 1H), 6.73 (d, J =
8.6 Hz, 1H), 2.95-2.40 (m, 2H), 1.86 (s, 3H), 1.78 (t, J = 6.7 Hz,
2H), 1.47-1.17 (m, 6H). 480.2 [M + H].sup.+ +++ ##STR00081##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.08 (d, J = 2.5 Hz,
1H), 7.63 (d, J = 8.6 Hz, 1H), 7.57 (ddd, J = 8.6, 2.5, 0.6 Hz,
1H), 7.27-7.22 (m, 2H), 7.04 (d, J = 2.4 Hz, 1H), 6.96 (ddd, J =
8.7, 2.5, 0.7 Hz, 1H), 6.86 (dt, J = 8.1, 0.6 Hz, 1H), 6.72 (d, J =
8.6 Hz, 1H), 2.75 (t, J = 6.8 Hz, 2H), 1.85 (s, 3H), 1.78 (t, J =
6.7 Hz, 2H), 1.32-1.30 (m, 6H). 496.5 [M + H].sup.+ +++
##STR00082## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.17 (d, J
= 8.4 Hz, 1H), 7.65 (d, J = 2.0 Hz, 1H), 7.49 (dd, J = 8.5, 2.1 Hz,
1H), 7.29-7.19 (m, 2H), 7.03 (d, J = 2.4 Hz, 1H), 7.00- 6.93 (m,
2H), 6.73 (d, J = 8.6 Hz, 1H), 2.75 (t, J = 6.8 Hz, 2H), 1.86 (s,
3H), 1.79 (t, J = 6.7 Hz, 2H), 1.32 (s, 6H). 496.5 [M + H].sup.+
+++ ##STR00083## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.21
(d, J = 2.7 Hz, 1H), 7.66 (dd, J = 8.9, 2.7 Hz, 1H), 7.29-7.16 (m,
3 H), 7.01 (d, J = 2.4 Hz, 1H), 6.96 (dd, J = 8.6, 2.5 Hz, 1H),
6.78 (dd, J = 8.1, 0.7 Hz, 1H), 6.73 (d, J = 8.6 Hz, 1H), 4.13 (s,
3H), 2.75 (t, J = 6.7 Hz, 2H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz,
2H), 1.32 (s, 6H). 492.3 [M + H].sup.+ +++ ##STR00084## .sup.1H NMR
(400 MHz, Chloroform-d) .delta. 7.47 (t, J = 7.7 Hz, 1H), 7.41-
7.30 (m, 3H), 7.22-7.16 (m, 2H), 7.05 (d, J = 2.4 Hz, 1H), 6.97
(dd, J = 8.6, 2.4 Hz, 1H), 6.85 (d, J = 8.8 Hz, 1H), 6.71 (d, J =
8.6 Hz, 1H), 3.71 (s, 2H), 2.74 (t, J = 6.8 Hz, 2H), 1.84 (s, 3H),
1.77 (t, J = 6.7 Hz, 2H), 1.31 (d, J = 1.8 Hz, 6H). 476.6 [M +
H].sup.+ +++ ##STR00085## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.12 (d, J = 2.1 Hz, 1H), 7.54 (dd, J = 8.2, 2.3 Hz, 1H),
7.41 (d, J = 8.2 Hz, 1H), 7.12-7.02 (m, 3H), 7.00 (dd, J = 8.6, 2.4
Hz, 1H), 6.80 (d, J = 7.8 Hz, 1H), 6.71 (d, J = 8.6 Hz, 1H), 2.75
(t, J = 6.8 Hz, 2H), 2.69 (s, 3H), 2.35 (s, 3H), 1.84 (s, 3H), 1.77
(t, J = 6.7 Hz, 2H), 1.37-1.27 (m, 6H). 456.0 [M + H].sup.+ +++
##STR00086## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.19 (t, J
= 1.9 Hz, 1H), 8.13- 8.06 (m, 1H), 7.76-7.69 (m, 1H), 7.61 (t, J =
7.9 Hz, 1H), 7.14-7.03 (m, 3H), 7.03-6.97 (m, 1H), 6.86- 6.77 (m,
1H), 6.71 (d, J = 8.6 Hz, 1H), 2.75 (t, J = 6.8 Hz, 2H), 2.36 (d, J
= 0.9 Hz, 3H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.31 (d, J =
1.8 Hz, 6H). 442.5 [M + H].sup.+ +++ ##STR00087## .sup.1H NMR (400
MHz, Chloroform-d) .delta. 8.26-8.20 (m, 2H), 7.61 (d, J = 8.6 Hz,
2H), 7.08 (dt, J = 6.3, 3.4 Hz, 3H), 6.98 (dd, J = 8.6, 2.5 Hz,
1H), 6.92 (d, J = 8.5 Hz, 1H), 6.71 (d, J = 8.6 Hz, 1H), 2.75 (t, J
= 6.8 Hz, 2H), 2.36 (s, 3H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz,
2H), 1.31 (d, J = 1.7 Hz, 6H). 442.5 [M + H].sup.+ +++ ##STR00088##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.17 (d, J = 2.7 Hz,
1H), 7.73 (dd, J = 8.8, 2.7 Hz, 1H), 7.51 (dd, J = 8.8, 1.4 Hz,
1H), 7.29-7.23 (m, 2H), 7.07-7.00 (m, 1H), 6.97 (dd, J = 8.6, 2.5
Hz, 1H), 6.90 (dd, J = 8.2, 0.6 Hz, 1H), 6.73 (d, J = 8.6 Hz, 1H),
2.74 (d, J = 6.8 Hz, 2H), 1.86 (s, 3H), 1.78 (s, 2H), 1.32 (d, J =
1.0 Hz, 6H). 546.6 [M + H].sup.+ +++ ##STR00089## .sup.1H NMR (400
MHz, Chloroform-d) .delta. 7.49-7.41 (m, 2H), 7.41-7.34 (m, 2H),
7.20 (dq, J = 4.3, 2.1 Hz, 2H), 1.04 (d, J = 2.3 Hz, 1H), 6.96
(ddd, J = 8.6, 1.9, 1.2 Hz, 1H), 6.85 (d, J = 8.9 Hz, 1H), 6.71 (d,
J = 8.5 Hz, 1H), 3.71 (s, 2H), 2.74 (t, J = 6.7 Hz, 2H), 1.84 (s,
3H), 1.78 (t, J = 6.7 Hz, 2H), 1.31 (d, J = 1.8 Hz, 6H). 476.2 [M +
H].sup.+ +++ ##STR00090## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.18 (s, 1H), 7.44-7.35 (m, 2H), 7.28 (dd, J = 8.4, 2.1 Hz,
1H), 7.25 (d, J = 2.1 Hz, 1H), 7.04-7.00 (m, 2H), 6.95 (dd, J =
8.6, 2.4 Hz, 1H), 6.73 (d, J = 8.6 Hz, 1H), 2.75 (t, J = 6.8 Hz,
2H), 1.86 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J = 1.1 Hz,
6H). 480.5 [M + H].sup.+ +++ ##STR00091## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.19 (t, J = 1.8 Hz, 1H), 8.10 (dt, J = 7.8,
1.4 Hz, 1H), 7.72 (ddd, J = 7.9, 2.1, 1.2 Hz, 1H), 7.62 (t, J = 7.9
Hz, 1H), 7.11-7.04 (m, 3H), 7.01 (dd, J = 8.6, 2.5 Hz, 1H) 6.83 (d,
J = 7.9 Hz, 1H), 6.71 (d, J = 8.6 Hz, 1H), 2.75 (t, J = 6.7 Hz,
2H), 2.36 (s, 3H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.31 (d,
J = 1.8 Hz, 6H). 442.6 [M + H].sup.+ +++ ##STR00092## .sup.1H NMR
(400 MHz, Chloroform-d) .delta. 8.28-8.17 (m, 1H), 7.40-7.35 (m,
2H), 7.27-7.23 (m, 2H), 7.06 (d, J = 2.4 Hz, 1H), 6.97 (dd, J =
8.6, 2.5 Hz, 1H), 6.95-6.89 (m, 1H), 6.73 (d, J = 8.6 Hz, 1H), 2.76
(t, J = 6.8 Hz, 2H), 2.71 (s, 3H), 1.86 (s, 3H), 1.78 (t, J = 6.7
Hz, 2H), 1.32 (d, J = 1.6 Hz, 6H). 476.2 [M + H].sup.+ +++
##STR00093## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 7.39-7.30
(m, 4H), 7.22-7.15 (m, 2H), 7.05 (d, J = 2.4 Hz, 1H), 6.99-6.94 (m,
1H), 6.85-6.80 (m, 1H), 6.71 (d, J = 8.6 Hz, 1H), 3.02 (t, J = 7.7
Hz, 2H), 2.81-2.68 (m, 4H), 1.83 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H),
1.31 (d, J = 1.9 Hz, 6H). 490.2 [M + H].sup.+ +++ ##STR00094##
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.05 (d, J = 2.3, Hz, 1
H), 7.56 (dd, J = 8.5, 2.3 Hz, 1 H), 7.46 (d, J = 8.2 Hz, 1 H),
7.23-7.20 (m, 2 H), 7.05 (d, J = 2.4, Hz, 1 H), 6.98 (dd, J = 8.6,
2.3 Hz, 1 H) 6.85-6.83 (m, 1H), 6.73 (d, J = 8.6 Hz, 1 H), 3.11 (q,
J = 7.4 Hz, 2 H), 3.07 (bs, 1 H), 2.76 (t, J = 7.6 Hz, 2 H), 1.84
(s, 3 H), 1.79 (t, J = 6.6 Hz, 2 H), 1.31- 1.25 (m, 9 H). 490.2 [M
+ H].sup.+ +++ ##STR00095## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.30 (dd, J = 8.8, 0.6 Hz, 1H), 7.31-7.19 (m, 4H),
7.07-7.02 (m, 1H), 7.00 (dt, J = 8.4, 0.5 Hz, 1H), 6.95 (ddd, J =
8.7, 2.5, 0.7 Hz, 1H), 6.73 (d, J = 8.6 Hz, 1H), 4.09 (s, 3H),
2.86-2.52 (m, 2H), 1.86 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H),
1.34-1.29 (m, 6H). 492.2 [M + H].sup.+ +++ ##STR00096## .sup.1H NMR
(400 MHz, Chloroform-d) .delta. 7.49-7.39 (m, 1H), 7.25 (ddd, J =
7.5, 5.3, 3.1 Hz, 3H), 7.20-7.18 (m, 1H), 7.05 (d, J = 2.4 Hz, 1H),
6.97 (dd, J = 8.6, 2.5 Hz, 1H), 6.85- 6.79 (m, 1H), 6.72 (d, J =
8.6 Hz, 1H), 3.00 (t, J = 7.7 Hz, 2H), 2.73 (dt, J = 17.0, 7.2 Hz,
4H), 1.84 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.31 (d, J = 1.9 Hz,
6H). 490.2 [M + H].sup.+ +++ ##STR00097## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.06-7.99 (m, 2H), 7.51-7.35 (m, 2H),
7.20-7.07 (m, 2H), 7.00- 6.82 (m, 2H), 6.71 (s, 1H), 6.66 (d, J =
8.9 Hz, 1H), 5.11-4.75 (m, 2H), 2.74 (t, J = 6.7 Hz, 2H), 1.82 (s,
3H), 1.78 (t, J = 6.7 Hz, 2H), 1.31 (s, 6H). 476.2 [M + H].sup.+
+++ ##STR00098## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 7.47
(t, J = 7.8 Hz, 1H), 7.42- 7.29 (m, 3H), 7.22-7.16 (m, 2H), 7.05
(dd, J = 2.5, 1.2 Hz, 1H), 7.00- 6.94 (m, 1H), 6.84 (dt, J = 8.0,
1.0 Hz, 1H), 6.72 (d, J = 8.6 Hz, 1H), 3.80 (d, J = 7.4 Hz, 1H),
2.75 (t, J = 6.7 Hz, 2H), 1.84 (d, J = 1.0 Hz, 3H), 1.78 (t, J =
6.7 Hz, 2H), 1.55 (d, J = 7.0 Hz, 3H), 1.31 (d, J = 1.8 Hz, 6H).
490.6 [M + H].sup.+ +++ ##STR00099## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.05 (d, J = 8.2 Hz, 2H), 7.34 (d, J = 8.1
Hz, 2H), 7.19-7.12 (m, 2H), 6.99 (d, J = 2.4 Hz, 1H), 6.91 (dd, J =
8.6, 2.5 Hz, 1H), 6.72 (d, J = 8.5 Hz, 1H), 6.67-6.60 (m, 1H),
5.09-4.89 (m, 2H), 2.74 (t, J = 6.8 Hz, 2H), 1.81 (s, 3H), 1.78 (t,
J = 6.7 Hz, 2H), 1.32 (s, 6H). 476.2 [M + H].sup.+ +++ ##STR00100##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.16 (dd, J = 7.8, 1.5
Hz, 1H), 7.53-7.43 (m, 1H), 7.42-7.33 (m, 1H), 7.21-7.12 (m, 2H),
7.07- 6.99 (m, 2H), 6.95 (dd, J = 8.6, 2.5 Hz, 1H), 6.72 (d, J =
8.5 Hz, 1H), 6.66 (dd, J = 8.3, 0.6 Hz, 1H), 5.44 (d, J = 3.8 Hz,
2H), 2.75 (t, J = 6.7 Hz, 2H), 1.84 (s, 3H), 1.78 (t, J = 6.7 Hz,
2H), 1.32 (s, 6H). 476.2 [M + H].sup.+ +++ ##STR00101## .sup.1H NMR
(400 MHz, Chloroform-d) .delta. 7.53-7.42 (m, 3H), 7.34-7.28 (m,
1H), 7.22-7.17 (m, 2H), 7.08- 7.03 (m, 1H), 6.97 (dd, J = 8.6, 2.5
Hz, 1H), 6.82 (dd, J = 8.1, 0.8 Hz, 1H), 6.72 (d, J = 8.6 Hz, 1H),
2.75 (t, J = 6.8 Hz, 2H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H),
1.63 (s, 6H), 1.32 (d, J = 2.1 Hz, 6H). 504.2 [M + H].sup.+ +++
##STR00102## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.17 (d, J
= 2.4 Hz, 1H), 7.45- 7.37 (m, 1H), 7.20-7.10 (m, 2H), 7.03-6.96 (m,
2H), 6.90-6.83 (m, 1H), 6.76-6.63 (m, 2H), 4.98- 4.82 (m, 2H), 4.06
(s, 3H), 2.75 (t, J = 6.7 Hz, 2H), 1.81-1.74 (m, 5H), 1.32 (s, 6H).
528.2 [M + Na].sup.+ +++ ##STR00103## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 8.08 (s, 1H), 8.02 (d, J = 7.7 Hz, 1H),
7.60-7.37 (m, 2H), 7.11 (s, 1H), 7.06-6.96 (m, 2H), 6.92- 6.85 (m,
1H), 6.77-6.65 (m, 1H), 6.41 (t, J = 8.6 Hz, 1H), 5.87 (m, J = 7.0
Hz, 1H), 2.76 (d, J = 7.0 Hz, 2H), 1.87 (dd, J = 7.1, 4.0 Hz, 2H),
1.81-1.74 (m, 6H), 1.32 (d, J = 2.3 Hz, 6H). 512.2 [M + Na].sup.+
+++ ##STR00104## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 7.92
(d, J = 6.8 Hz, 1H), 7.44 (s, 1H), 7.22-7.08 (m, 3H), 6.97 (s, 1H),
6.89 (d, J = 10.7 Hz, 1H), 6.68 (dd, J = 16.5, 8.4 Hz, 2H), 5.06-
4.79 (m, 2H), 2.73 (t, J = 6.7 Hz, 3H), 1.81-1.68 (m, 5H), 1.31 (s,
6H). 516.3 [M + Na].sup.+ +++ ##STR00105## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 7.93 (s, 1H), 7.33-7.27 (m, 1H), 7.22 (d, J =
7.0 Hz, 1H), 7.18- 7.12 (m, 2H), 6.99 (d, J = 4.2 Hz, 1H),
6.92-6.86 (m, 1H), 6.72- 6.64 (m, 2H), 5.03-4.78 (m, 2H), 2.74 (t,
J = 6.7 Hz, 2H), 2.60 (s, 3H), 1.80 (m, 5H), 1.31 (s, 6H). 512.3 [M
+ Na].sup.+ +++ ##STR00106## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 7.92 (s, 1H), 7.25-7.13 (m, 3H), 7.00 (d, J = 4.5 Hz, 1H),
6.97- 6.90 (m, 2H), 6.69 (dd, J = 17.2, 8.3 Hz, 2H), 5.35 (d, J =
5.1 Hz, 2H), 2.74 (t, J = 6.8 Hz, 2H), 2.36 (s, 3H), 1.81 (s, 3H),
1.78 (t, J = 6.7 Hz, 2H), 1.32 (s, 6H). 490.3 [M + H].sup.+ +++
##STR00107## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 7.42 (t, J
= 8.1 Hz, 1H), 7.22- 7.16 (m, 2H), 7.08-7.02 (m, 2H), 7.01-6.92 (m,
3H), 6.86 (dd, J = 9.0, 0.6 Hz, 1H), 6.71 (d, J = 8.6 Hz, 1H), 4.66
(s, 2H), 2.74 (t, J = 6.7 Hz, 2H), 1.83 (s, 3H), 1.77 (t, J = 6.7
Hz, 2H), 1.31 (d, J = 1.8 Hz, 6H). 492.5 [M + H].sup.+ +++
##STR00108## .sup.1H NMR (400 MHz, Methanol-d.sub.4) .delta. 7.77
(dd, J = 9.5, 2.8 Hz, 1H), 7.33-7.17 (m, 2H), 7.12-7.03 (m, 1H),
6.99 (d, J = 2.4 Hz, 1H), 6.94 (d, J = 8.4 Hz, 1H), 6.87- 6.80 (m,
2H), 6.67 (d, J = 8.6 Hz, 1H), 5.38 (s, 2H), 2.76 (t, J = 6.6 Hz,
2H), 1.85-1.75 (m, 5H), 1.29 (d, J = 1.9 Hz, 6H). 494.3 [M +
H].sup.+ +++ ##STR00109## .sup.1H NMR (400 MHz, Methanol-d.sub.4)
.delta. 7.58 (d, J = 2.6 Hz, 1H), 7.24- 7.19 (m, 3H), 7.04-6.97 (m,
2H), 6.97-6.90 (m, 1H), 6.82 (d, J = 8.6 Hz, 1H), 6.67 (d, J = 8.5
Hz, 1H), 5.33 (s, 2H), 3.80 (s, 3H), 2.76 (t, J = 6.8 Hz, 2H),
1.88-1.67 (m, 5H), 1.29 (d, J = 2.0 Hz, 6H). 506.4 [M + H].sup.+
+++ ##STR00110## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 7.32
(d, J = 8.8 Hz, 2H), 7.23- 7.16 (m, 2H), 7.06-7.01 (m, 3H),
6.97-6.88 (m, 1H), 6.80-6.74 (m, 1H), 6.72 (d, J = 8.6 Hz, 1H),
4.69 (s, 2H), 2.74 (s, 2H), 1.84 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H),
1.31 (d, J = 1.8 Hz, 6H). 492.5 [M + H].sup.+ +++ ##STR00111##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 7.37-7.28 (m, 4H), 7.20
(dqd, J = 4.2, 2.2, 0.6 Hz, 2H), 7.05 (d, J = 2.4 Hz, 1H), 6.96
(ddd, J = 8.8, 2.5, 0.8 Hz, 1H), 6.81 (dd, J = 8.9, 0.6 Hz, 1H),
6.72 (d,
J = 8.5 Hz, 1H), 2.79-2.65 (m, 4H), 2.42 (t, J = 7.3 Hz, 2H), 2.00
(p, J = 7.4 Hz, 2H), 1.84 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32
(d, J = 2.0 Hz, 6H). 504.6 [M + H].sup.+ +++ ##STR00112## .sup.1H
NMR (400 MHz, Chloroform-d) .delta. 8.23 (d, J = 2.7 Hz, 1H), 7.64
(ddd, J = 8.9, 2.8, 0.6 Hz, 1H), 7.25- 7.15 (m, 3H), 7.01 (d, J =
2.4 Hz, 1H), 6.96 (ddd, J = 8.7, 2.5, 0.7 Hz, 1H), 6.78 (dt, J =
8.1, 0.7 Hz, 1H), 6.73 (d, J = 8.5 Hz, 1H), 4.40 (q, J = 7.0 Hz,
2H), 2.75 (t, J = 6.7 Hz, 2H), 1.85 (s, 3H), 1.78 (t, J = 6.7 Hz,
2H), 1.61 (t, J = 7.0 Hz, 3H), 1.32 (s, 6H). 506.4 [M + H].sup.+
+++ ##STR00113## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.25
(d, J = 2.7 Hz, 1H), 7.63 (dd, J = 8.8, 2.8 Hz, 1H), 7.25-7.16 (m,
3H), 7.06-6.92 (m, 2H), 6.80 (d, J = 9.0 Hz, 1H), 6.72 (d, J = 8.5
Hz, 1H), 4.93 (p, J = 6.1 Hz, 1H), 2.75 (t, J = 6.7 Hz, 2H), 1.84
(s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.53 (dd, J = 6.0, 1.0 Hz, 6H),
1.32 (s, 6H). 520.4 [M + H].sup.+ +++ ##STR00114## .sup.1H NMR (400
MHz, Chloroform-d) .delta. 8.19-8.05 (m, 1H), 7.39 (dd, J = 8.4,
2.3 Hz, 1H), 7.23-7.16 (m, 2H), 7.03-6.96 (m, 2H), 6.92 (dd, J =
8.5, 2.5 Hz, 1H), 6.72 (d, J = 8.6 Hz, 1H), 6.65 (d, J = 8.1 Hz,
1H), 5.43-5.24 (m, 2H), 2.74 (t, J = 6.8 Hz, 2H), 2.40 (br s, 1H),
1.88- 1.64 (m, 5H), 1.32 (s, 6H). 510.3 [M + H].sup.+ +++
##STR00115## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 7.97 (d, J
= 7.9 Hz, 1H), 7.24 (s, 1H), 7.18-7.13 (m, 2H), 7.10 (s, 1H), 6.98
(s, 1H), 6.91 (d, J = 8.6 Hz, 1H), 6.71 (d, J = 8.6 Hz, 1H), 6.63
(d, J = 8.8 Hz, 1H), 5.09- 4.65 (m, 2H), 2.77-2.70 (m, 2H), 2.57
(d, J = 0.8 Hz, 3H), 1.87- 1.61 (m, 5H), 1.32 (s, 6H). 490.4 [M +
H].sup.+ +++ ##STR00116## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.05-8.00 (m, 1H), 7.19-7.10 (m, 3H), 7.06-7.02 (m, 1H),
6.97- 6.92 (m, 1H), 6.82-6.79 (m, 1H), 6.74-6.64 (m, 1H), 5.38 (d,
J = 21.9 Hz, 2H), 2.79-2.74 (m, 2H), 2.22 (s, 3H), 1.83 (s, 3H),
1.79- 1.74 (m, 2H), 1.32 (d, J = 3.2 Hz, 6H). 490.4 [M + H].sup.+
+++ ##STR00117## .sup.1H NMR (400 MHz, Chloroform-d) .delta.
7.26-7.25 (m, 1H), 7.25-7.21 (m, 1H), 7.20 (d, J = 9.1 Hz, 1H),
7.18-7.13 (m, 1H), 6.99 (d, J = 7.6 Hz, 1H), 6.91 (d, J = 8.4 Hz,
1H), 6.85 (d, J = 8.1 Hz, 2H), 6.68 (d, J = 8.1 Hz, 1H), 5.11-4.71
(m, 2H), 2.71-2.66 (m, 2H), 2.44 (s, 3H), 1.77-1.73 (m, 5H), 1.30
(s, 6H). 512.3 [M + Na].sup.+ +++ ##STR00118## .sup.1H NMR (400
MHz, Chloroform-d) .delta. 7.75 (t, J = 1.6 Hz, 1H), 7.65 (dd, J =
2.5, 1.3 Hz, 1H), 7.26-7.19 (m, 3H), 7.05 (d, J = 2.5 Hz, 1H), 6.99
(dd, J = 8.6, 2.5 Hz, 1H), 6.90- 6.84 (m, 1H), 6.73 (d, J = 8.6 Hz,
1H), 3.89 (s, 3H), 2.76 (t, J = 6.7 Hz, 2H), 1.86 (s, 3H), 1.78 (t,
J = 6.7 Hz, 2H), 1.32 (d, J = 1.6 Hz, 6H). 492.5 [M + H].sup.+ +++
##STR00119## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 7.95 (dd,
J = 1.8, 0.8 Hz, 2H), 7.50 (dq, J = 2.0, 0.9 Hz, 1H), 7.22 (dt, J =
4.3, 2.4 Hz, 2H), 7.06 (d, J = 2.4 Hz, 1H), 6.99 (dd, J = 8.6, 2.4
Hz, 1H), 6.83 (d, J = 9.0 Hz, 1H), 6.73 (d, J = 8.6 Hz, 1H), 2.76
(t, J = 6.8 Hz, 2H), 2.48 (s, 3H), 1.86 (s, 3H), 1.78 (t, J = 6.7
Hz, 2H), 1.32 (d, J = 1.6 Hz, 6H). 476.3 [M + H].sup.+ +++
##STR00120## .sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.09 (d, J
= 2.1 Hz, 1H), 7.94 (d, J = 8.5 Hz, 1H), 7.79 (ddd, J = 8.5, 2.0,
0.8 Hz, 1H), 7.31-7.22 (m, 2H), 7.03 (d, J = 2.4 Hz, 1H), 7.01-
6.90 (m, 2H), 6.73 (d, J = 8.6 Hz, 1H), 2.75 (t, J = 6.8 Hz, 2H),
1.87 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (s, 6H). 530.3 [M +
H].sup.+ +++ ##STR00121## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 7.86 (d, J = 7.7 Hz, 1H), 7.20- 7.08 (m, 4H), 7.03 (d, J =
2.4 Hz, 1H), 6.94 (dd, J = 8.6, 2.5 Hz, 1H), 6.72 (d, J = 8.6 Hz,
1H), 6.54 (d, J = 8.3 Hz, 1H), 4.98 (d, J = 5.6 Hz 2H), 2.75 (t, J
= 6.8 Hz, 2H), 2.62 (s, 3H), 1.83 (s, 3H), 1.78 (t, J = 6.8 Hz,
2H), 1.32 (s, 6H). 490.3 [M + H].sup.+ +++ ##STR00122## .sup.1H NMR
(400 MHz, Chloroform-d) .delta. 7.63 (d, J = 7.2 Hz, 1H), 7.35-
7.28 (m, 2H), 7.25-7.18 (m, 1H), 7.17 (dd, J = 2.0, 1.1 Hz, 1H),
6.84 (dd, J = 13.4, 5.0 Hz, 3H), 6.67 (d, J = 8.5 Hz, 1H), 5.11 (s,
2H), 2.74- 2.47 (m, 2H), 2.18 (d, J = 1.0 Hz, 3H), 1.76 (t, J = 6.6
Hz, 2H), 1.72 (d, J = 1.1 Hz, 3H), 1.30 (s, 6H). 490.4 [M +
H].sup.+ +++ ##STR00123## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 7.48-7.41 (m, 3H), 7.30-7.23 (m, 2H), 7.20-7.12 (m, 2H),
7.03 (dd, J = 15.9, 2.5 Hz, 1H), 6.94 (ddd, J = 12.7, 8.5, 2.5 Hz,
1H), 6.70 (dd, J = 8.6, 7.0 Hz, 1H), 6.55 (dd, J = 26.1, 8.3 Hz,
1H), 3.74- 3.30 (m, 2H), 2.72 (q, J = 6.7 Hz, 2H), 1.84 (s, 3H),
1.77 (q, J = 6.8 Hz, 2H), 1.31 (d, J = 2.0 Hz, 6H). 476.3 [M +
H].sup.+ +++ ##STR00124## .sup.1H NMR (400 MHz, Chloroform-d)
.delta. 8.23 (d, J = 2.7 Hz, 1H), 7.63 (dd, J = 8.8, 2.7 Hz, 1H),
7.23-7.17 (m, 3H), 7.02 (d, J = 2.4 Hz, 1H), 6.97 (dd, J = 8.6, 2.4
Hz, 1H), 6.81- 6.75 (m, 1H), 6.73 (d, J = 8.5 Hz, 1H), 4.08 (d, J =
6.4 Hz, 2H), 2.75 (t, J = 6.7 Hz, 2H), 2.27 (dt, J = 13.3, 6.7 Hz,
1H), 1.84 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (s, 6H), 1.28 (s,
2H), 1.12 (d, J = 6.7 Hz, 6H). 534.4 [M + H].sup.+ +++ ##STR00125##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.09 (d, J = 8.4 Hz,
1H), 7.34 (dd, J = 8.4, 2.1 Hz, 1H), 7.24-7.16 (m, 2H), 7.04-6.89
(m, 3H), 6.75 (d, J = 8.5 Hz, 1H), 6.65 (d, J = 9.1 Hz, 1H), 5.45
(d, J = 17.9 Hz, 1H), 5.32 (d, J = 17.9 Hz, 1H), 2.75 (t, J = 6.7
Hz, 2H), 1.84 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J = 1.4
Hz, 6H). 532.3 [M + Na].sup.+ +++ ##STR00126## .sup.1H NMR (400
MHz, Chloroform-d) .delta. 8.08 (dt, J = 12.0, 1.6 Hz, 2H), 7.71
(t, J = 1.9 Hz, 1H), 7.28-7.26 (m, 1H), 7.24 (t, J = 1.6 Hz, 1H),
7.04 (d, J = 2.4 Hz, 1H), 6.97 (dd, J = 8.6, 2.5 Hz, 1H), 6.88 (d,
J = 8.2 Hz, 1H), 6.73 (d, J = 8.5 Hz, 1H), 2.76 (t, J = 6.7 Hz,
2H), 1.86 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J = 1.2 Hz,
6H). 496.3 [M + H].sup.+ +++ ##STR00127## .sup.1H NMR (400 MHz,
Chloroform-d) .delta. 7.37 (s, 1H), 7.28-7.27 (m, 1H), 7.24-7.21
(m, 1H), 7.16 (d, J = 2.1 Hz, 1H), 7.05 (d, J = 7.7 Hz, 1H), 6.92
(d, J = 2.4 Hz, 1H), 6.86 (d, J = 8.4 Hz, 2H), 6.69 (d, J = 8.6 Hz,
1H), 4.96 (d, J = 2.0 Hz, 2H), 2.88-2.53 (m, 2H), 1.79-1.66 (m,
5H), 1.31 (s, 6H). 532.3 [M + Na].sup.+ +++ ##STR00128## .sup.1H
NMR (400 MHz, Chloroform-d) .delta. 7.42-7.31 (m, 2H), 7.31-7.27
(m, 1H), 7.18 (d, J = 8.5 Hz, 1H), 7.10 (d, J = 2.1 Hz, 1H), 6.94
(d, J = 8.1 Hz, 1H), 6.74 (d, J = 2.4 Hz, 1H), 6.69 (dd, J = 8.6,
2.5 Hz, 1H), 6.60 (d, J = 8.6 Hz, 1H), 4.96 (d, J = 3.4 Hz, 2H),
3.58 (s, 3H), 2.64 (q, J = 6.5 Hz, 2H), 1.73 (t, J = 6.7 Hz, 2H),
1.68 (s, 3H), 1.28 (s, 6H). 506.3 [M + H].sup.+ ++ ##STR00129##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 7.33 (t, J = 8.2 Hz,
2H), 7.21- 7.13 (m, 2H), 7.03-6.83 (m, 2H), 6.76-6.65 (m, 3H), 5.22
(s, 2H), 4.00 (s, 3H), 2.73 (t, J = 6.8 Hz, 2H), 1.80-1.73 (m, 5H),
1.31 (s, 6H). 506.3 [M + H].sup.+ +++ ##STR00130## .sup.1H NMR (400
MHz, Chloroform-d) .delta. 7.99 (t, J = 1.6 Hz, 1H), 7.81 (ddd, J =
8.4, 2.5, 1.4 Hz, 1H), 7.48 (ddd, J = 8.8, 2.4, 1.8 Hz, 1H), 7.28-
7.23 (m, 2H), 7.03 (dd, J = 2.4, 1.1 Hz, 1H), 6.97 (dd, J = 8.6,
2.5 Hz, 1H), 6.92 (dd, J = 8.3, 0.5 Hz, 1H), 6.73 (d, J = 8.5 Hz,
1H), 2.76 (t, J = 6.8 Hz, 2H), 1.86 (s, 3H), 1.79 (t, J = 6.7 Hz,
2H), 1.35-1.16 (m, 6H). 480.3 [M + H].sup.+ +++ ##STR00131##
.sup.1H NMR (400 MHz, Chloroform-d) .delta. 8.13 (d, J = 8.7 Hz,
1H), 7.22- 7.15 (m, 2H), 7.05 (s, 1H), 6.97 (d, J = 8.6 Hz, 1H),
6.81 (dd, J = 8.7, 2.7 Hz, 1H), 6.69 (dd, J = 13.3, 8.4 Hz, 2H),
6.49 (d, J = 2.6 Hz, 1H), 5.58-5.08 (m, 2H), 3.59 (s, 3H), 2.74 (t,
J = 6.8 Hz, 2H), 1.83 (s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J
= 1.6 Hz, 6H). 506.3 [M + H].sup.+ +++ ##STR00132## .sup.1H NMR
(400 MHz, Chloroform-d) .delta. 8.49-.8.29 (m, 2H), 8.04 (dd, J =
2.1, 1.5 Hz, 1H), 7.31-7.27 (m, 2H), 7.04-6.94 (m, 2H), 6.92- 6.71
(m, 2H), 2.76 (t, J = 6.8 Hz, 2H), 1.88 (s, 3H), 1.79 (t, J = 6.7
Hz, 2H), 1.32 (s, 6H). 487.3 [M + H].sup.+ ++ ##STR00133## .sup.1H
NMR (400 MHz, Chloroform-d) .delta. 8.01-7.96 (m, 1H), 7.55-7.48
(m, 1H), 7.24-7.20 (m, 2H), 7.05- 7.00 (m, 1H), 7.01-6.92 (m, 1H),
6.85-6.78 (m, 1H), 6.73 (d, J = 8.5 Hz, 1H), 3.96 (d, J = 0.7 Hz,
3H), 2.75 (dd, J = 7.2, 6.2 Hz, 2H), 2.41 (t, J = 0.7 Hz, 3H), 1.84
(s, 3H), 1.78 (t, J = 6.7 Hz, 2H), 1.32 (d, J = 0.7 Hz, 6H). 506.3
[M + H].sup.+ +++ ##STR00134## .sup.1H NMR (400 MHz, chloroform-d)
.delta. 7.58-7.51 (m, 2H), 7.47-7.41 (m, 2H), 7.22-7.16 (m, 2H),
7.03 (d, J = 2.4 Hz, 1H), 6.96 (dd, J = 8.6, 2.5 Hz, 1H), 6.85-6.80
(m, 1H), 6.70 (d, J = 8.6 Hz, 1H), 5.16 (s, 2H), 2.73 (t, J = 6.7
Hz, 2H), 1.83 (s, 3H), 1.77 (t, J = 6.7 Hz, 2H), 1.30 (d, J = 2.0
Hz, 6H) 516.5 +++ ##STR00135## .sup.1H NMR (400 MHz, chloroform-d)
.delta. 7.58-7.52 (m, 2H), 7.44-7.39 (m, 2H), 7.21-7.16 (m, 2H),
7.03 (d, J = 2.4 Hz, 1H), 6.98-6.93 (m, 1H), 6.85-6.80 (m, 1H),
6.70 (d, J = 8.6 Hz, 1H), 5.13 (s, 2H), 3.61 (s, 3H), 2.73 (t, J =
6.7 Hz, 2H), 1.82 (s, 3H), 1.76 (t, J = 6.7 Hz, 2H), 1.30 (d, J =
1.8 Hz, 6H). 530.5 +++
Biological Examples
Measuring Efficacy of Chemokine Modulators
[0165] Examples of In Vitro Assays--Reagents
[0166] MOLT-4 cells were obtained from the American Type Culture
Collection (Manassas, Va.) and cultured in RPMI tissue culture
medium supplemented with 10% fetal calf serum (FCS) in a humidified
5% CO2 incubator at 37.degree. C. Recombinant human chemokine
proteins TECK was obtained from R&D Systems (Minneapolis,
Minn.). ChemoTX.TM. chemotaxis microchambers were purchased from
Neuro Probe (Gaithersburg, Md.). CyQUANT.TM. cell proliferation
kits were purchased from Molecular Probes (Eugene, Oreg.). Calcium
indicator dye Fluo-4 AM was purchased from Molecular Devices
(Mountain View, Calif.).
[0167] In Vitro Assays
[0168] A variety of assays can be used to evaluate the compounds
provided herein, including signaling assays, chemotaxis (migration
assays), ligand binding assays, and other assays of cellular
response. Chemokine receptor signaling assays can be used to
measure the ability of a compound, such as a potential CCR(9)
antagonist, to block CCR(9) ligand- (e.g. TECK)-induced signaling.
Blocking such signaling can be useful in treating various diseases
such as inflammatory bowel diseases, an allergic disease,
psoriasis, atopic dermatitis, asthma, fibrotic diseases, graft
rejection, immune mediated food allergies, autoimmune diseases,
Celiac disease, rheumatoid arthritis, thymoma, thymic carcinoma,
leukemia, solid tumor, acute lymphocytic leukemia, melanoma,
primary sclerosing cholangitis, hepatitis or post-operative
ileus.
[0169] Chemotaxis assays can also be used to assess receptor
function and evaluate the compounds provided herein. These assays
are based on the functional migration of cells in vitro or in vivo
induced by an agent, and can be used to assess the binding and/or
effect on chemotaxis of ligands, inhibitors, or agonists. A variety
of chemotaxis assays are known in the art, and any suitable assay
can be used to evaluate the compounds of the present disclosure.
Examples of suitable assays include those described in
PCT/US97/15915; Springer et al., WO 94/20142; Berman et al.,
Immunol. Invest., 17:625-677 (1988); and Kavanaugh et al., J.
Immunol., 146:4149-4156 (1991)).
[0170] Evaluation of a Test Modulator in a Serum Chemotaxis
Assay
[0171] A serum chemotaxis assay was used to determine the efficacy
of potential receptor antagonists at blocking the migration
mediated through chemokine receptors, such as CCR(9). This assay
was performed using the ChemoTX.RTM. microchamber system with a
5-.mu.m pore-sized polycarbonate membrane. MOLT-4 cells were
collected by centrifugation at 400.times.g at room temperature,
then suspended at 50 million/ml in human serum, containing 50 mM
HEPES (final pH of 7.2). The compound being tested or an equivalent
volume of its solvent (DMSO) was then added to the cell/serum
mixture at a final DMSO concentration of 0.125% (v/v), and this
mixture was then incubated together at 37.degree. C. for one hour.
Separately, recombinant human TECK was diluted with chemotaxis
buffer (HBSS+0.1% BSA), generally spanning a range from 0.1 nM to
500 nM, after which 29 .mu.l of diluted chemokine was placed in the
lower wells of the ChemoTX.RTM. plate. The 5-.mu.m (pore size)
polycarbonate membrane was placed onto the plate, and 20 .mu.L of
the cell/compound mixture was transferred onto each well of the
membrane. The plates were incubated at 37.degree. C. for 90
minutes, after which the polycarbonate membranes were removed and 5
.mu.l of the DNA-intercalating agent CyQUANT (Invitrogen, Carlsbad,
Calif.) was added to the lower wells. The amount of fluorescence,
corresponding to the number of migrated cells, was measured using a
Spectrafluor Plus plate reader (TECAN, San Jose, Calif.).
[0172] The A.sub.2 values were calculated from the following
equation, comparing the efficacy of the test compound with that of
the DMSO-only control at equi-active chemokine levels:
Log(A.sub.2)=log [drug(M)]-log [(A'/A)-1]
where A reflects the potency of the agonist in the absence of
antagonist and A' reflects the potency of the agonist in the
presence of antagonist at a given concentration of drug
([drug(M)]).
[0173] In Vivo Efficacy Models for Human IBD
[0174] T cell infiltration into the small intestine and colon have
been linked to the pathogenesis of human inflammatory bowel
diseases which include Coeliac disease, Crohn's disease and
ulcerative colitis. Blocking trafficking of relevant T cell
populations to the intestine is believed to be an effective
approach to treat human IBD. CCR(9) is expressed on gut-homing T
cells in peripheral blood, elevated in patients with small bowel
inflammation such as Crohn's disease and Coeliac disease. CCR(9)
ligand TECK is expressed in the small intestine. It is thus
believed that this ligand-receptor pair plays a role in IBD
development by mediating migration of T cells to the intestine.
Several animal models exist and can be used for evaluating
compounds of interest, such as potential CCR(9) antagonists, for an
ability to affect such T cell migration and/or condition or
disease, which might allow efficacy predictions of antagonists in
humans.
[0175] Animal Models with Pathology Similar to Human Ulcerative
Colitis
[0176] A murine model described by Panwala and coworkers (Panwala
et al., J Immunol., 161(10):5733-44 (1998)) involves genetic
deletion of the murine multi-drug resistant gene (MDR). MDR
knockout mice (MDR-/-) are susceptible to developing a severe,
spontaneous intestinal inflammation when maintained under specific
pathogen-free facility conditions. The intestinal inflammation seen
in MDR-/- mice has a pathology similar to that of human
inflammatory bowel disease (IBD) and is defined by Th1 type T cells
infiltration into the lamina propria of the large intestine.
[0177] Another murine model was described by Davidson et al., J Exp
Med., 184(1):241-51(1986). In this model, the murine IL-10 gene was
deleted and mice rendered deficient in the production of
interleukin 10 (IL-10-/-). These mice develop a chronic
inflammatory bowel disease (IBD) that predominates in the colon and
shares histopathological features with human IBD.
[0178] Another murine model for IBD has been described by Powrie et
al., Int. Immunol., 5(11):1461-71 (1993), in which a subset of CD4+
T cells (called CD45RB(high)) from immunocompetent mice are
purified and adoptively transferred into immunodeficient mice (such
as C.B-17 scid mice). The animal restored with the CD45RBhighCD4+ T
cell population developed a lethal wasting disease with severe
mononuclear cell infiltrates in the colon, pathologically similar
with human IBD.
[0179] The TNF ARE(-/-) Model.
[0180] The role of TNF in Crohn's disease in human has been
demonstrated more recently by success of treatment using anti-TNF
alpha antibody by Targan et al., N. Engl. J Med., 337(15): 1029-35
(1997). Mice with aberrant production of TNF-alpha due to genetic
alteration in the TNF gene (ARE-/-) develop Crohn's-like
inflammatory bowel diseases (see Kontoyiannis et al., Immunity,
10(3):387-98 (1999)).
[0181] Examples of In Vivo Efficacy Assays
[0182] Evaluation of a Test Modulator in a CCR(9) Dependent T Cell
Trafficking Model
[0183] Single cell suspensions were prepared from spleens and lymph
nodes of OT-I Tg CD45.1 mice. 15.times.10.sup.6 total cells (about
3.times.10.sup.6 CD8 T cells) were injected into sex-matched
congenic CD45.2 C57BL/6n mice (8-10 weeks old). 24 hours later,
animals were immunized via oral gavage with 25 mg Ovalbumin protein
(Sigma-Aldrich, St. Louis, Mo.)+10 ug Cholera Toxin (Calbiochem,
San Diego, Calif.). CCR(9) antagonist compound 1.063 (Table 1) was
administered prior to oral ovalbumin in a time frame dictated by
their mouse pharmacokinetics and dosed throughout. Five days post
immunization, animals were euthanized, and small intestines were
harvested. Peyer's patches were removed and, after flushing with
PBS, the gut was opened on a wet square of Optima fabric
(Allegiance Healthcare). The mucosa was scraped with a scalpel and
then dissociated by stirring in 50 ml of medium containing 10%
newborn calf serum and DTT (1 mM) for 15 min at room temperature.
After centrifugation, pellets were resuspended in PBS containing
10% newborn calf serum, vortexed for 3 min, and rapidly passed
through a glass wool column (1.6 g packed in a 20-ml syringe;
Fisher Scientific). IEL were further purified on a Ficoll-Paque
gradient and stained with mAbs for flow cytometry analysis.
Transferred OT-1 Tg CD45.1 T cells were detected and quantified by
flow cytometry. In this model treatment with a compound of the
disclosure resulted in a significant reduction in the frequency of
OT-1 Tg CD45.1 T cells that traffic to the small intestine in
response to antigen.
[0184] Evaluation of a Test Modulator in a Model of Inhibition of
HIV Spread
[0185] In the bone marrow/liver/thymus, or "BLT" mouse, non-obese
diabetic (NOD)/SCID mice (which lack endogenous T and B cells) are
surgically implanted with fetal thymic and liver organoids, as in
the SCID-hu system. The mice are then sub-lethally irradiated and
transplanted with autologous CD34.sup.+ stem cells obtained from
fetal liver which take up residence in the murine bone marrow,
effectively receiving a human bone marrow transplant and resulting
in a range of human cells in peripheral blood, including mature T
and B lymphocytes, monocytes, macrophages, and dendritic cells, all
of which show extensive infiltration of organs and tissues
including liver, lung, and gastrointestinal tract. Following
transplantation, a compound of the disclosure is administered to
transplanted mice to inhibit the trafficking of human cells to the
gastrointestinal tract, a major source of T cell/HIV interaction.
Compound efficacy is measured as a reduction in blood viral load by
standard techniques.
Evaluation of a Test Modulator in a Model of Arthritis
[0186] A 17-day study of type II collagen-induced arthritis is
conducted to evaluate the effects of a modulator on
arthritis-induced clinical ankle swelling. Rat collagen-induced
arthritis is an experimental model of polyarthritis that has been
widely used for preclinical testing of numerous anti-arthritic
agents (see Trentham et al., J. Exp. Med. 146(3):857-868 (1977),
Bendele et al., Toxicologic Pathol. 27:134-142 (1999), Bendele et
al., Arthritis. Rheum. 42:498-506 (1999)). The hallmarks of this
model are reliable onset and progression of robust, easily
measurable polyarticular inflammation, marked cartilage destruction
in association with pannus formation and mild to moderate bone
resorption and periosteal bone proliferation.
[0187] Female Lewis rats (approximately 0.2 kilograms) are
anesthetized with isoflurane and injected with Freund's Incomplete
Adjuvant containing 2 mg/mL bovine type II collagen at the base of
the tail and two sites on the back on days 0 and 6 of this 17-day
study. The test modulator is dosed daily by sub-cutaneous injection
from day 9 to day 17 at a dose of 100 mg/kg and a volume of 1 mL/kg
in the following vehicle (24.5% Cremaphore EL, 24.5% common oil, 1%
Benzylalcohol and 50% Distilled water). Caliper measurements of the
ankle joint diameter are taken daily, and reducing joint swelling
is taken as a measure of efficacy.
[0188] Evaluation of a Test Modulator in a Mouse Model of
Asthma
[0189] This example describes a procedure to evaluate the efficacy
of antagonists for treatment of asthma. An animal model of asthma
can be induced by sensitizing rodents to an experimental antigen
(e.g. OVA) by standard immunization, and subsequently introducing
that same antigen into the rodents lung by aerosolization. Three
series of rodent groups, comprising 10 rodents per goup, are
actively sensitized on Day 0 by a single i.p. injection with 100 ug
OVA in phosphate-buffered saline (PBS), along with an adjuvant e.g.
aluminum hydroxide. At 11 days after sensitization, the animals are
placed in a Plexiglas chamber and challenged with aerosolized OVA
(1%) for 30 minutes using the ultrasonic nebulizer (De Vilbliss).
One series of mice additionally receives PBS and Tween 0.5% i.p. at
the initial sensitization, and at different dosing schedules
thereafter, up until the aerosolized OVA challenge. A second series
consists of groups of mice receiving different doses of the CCR4
antagonist given either intraperitoneally, intra-venously,
sub-cutaneously, intra-muscularly, orally, or via any other mode of
administration at the initial sensitization, and at different
dosing schedules thereafter, up until the aerosolized OVA
challenge. A third series of mice, serving as positive control,
consists of groups treated with either mouse IL-10 i.p., anti-IL4
antibodies i.p., or anti-IL5 antibodies i.p. at the initial
sensitization, and at different dosing schedules thereafter, up
until the aerosolized OV A challenge. Animals are subsequently
analyzed at different time points after the aerosolized OVA
challenge for pulmonary function, cellular infiltrates in
bronchoalveolar lavage (BAL), histological examination of lungs,
and measurement of serum OVA specific IgE titers.
[0190] It is therefore intended that the foregoing detailed
description be regarded as illustrative rather than limiting, and
that it be understood that it is the following claims, including
all equivalents, that are intended to define the spirit and scope
of this disclosure.
* * * * *