U.S. patent application number 11/385630 was filed with the patent office on 2006-10-12 for thienopyridinone derivatives as macrophage migration inhibitory factor inhibitors.
Invention is credited to Timothy James Davis, Sunil Kumar K.C., Jagadish Sircar, Wenbin Ying.
Application Number | 20060229314 11/385630 |
Document ID | / |
Family ID | 36649727 |
Filed Date | 2006-10-12 |
United States Patent
Application |
20060229314 |
Kind Code |
A1 |
Sircar; Jagadish ; et
al. |
October 12, 2006 |
Thienopyridinone derivatives as macrophage migration inhibitory
factor inhibitors
Abstract
Inhibitors of macrophage migration inhibitory factor having a
thienopyridinone backbone are provided which have utility in the
treatment of a variety of disorders, including the treatment of
pathological conditions associated with macrophage migration
inhibitory factor activity. The inhibitors of macrophage migration
inhibitory factor have the following structures: ##STR1## including
forms such as stereoisomers, free forms, pharmaceutically
acceptable salts or esters thereof, solvates, or combinations of
such forms, wherein n, R.sub.1, R.sub.2, R.sub.3, X, and Y are as
defined herein. Compositions comprising an inhibitor of macrophage
migration inhibitory factor in combination with a pharmaceutically
acceptable carrier are also provided, as well as methods for use of
the same.
Inventors: |
Sircar; Jagadish; (San
Diego, CA) ; K.C.; Sunil Kumar; (San Diego, CA)
; Davis; Timothy James; (San Diego, CA) ; Ying;
Wenbin; (San Diego, CA) |
Correspondence
Address: |
KNOBBE MARTENS OLSON & BEAR LLP
2040 MAIN STREET
FOURTEENTH FLOOR
IRVINE
CA
92614
US
|
Family ID: |
36649727 |
Appl. No.: |
11/385630 |
Filed: |
March 20, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60665236 |
Mar 24, 2005 |
|
|
|
60733657 |
Nov 4, 2005 |
|
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Current U.S.
Class: |
514/253.04 ;
544/362 |
Current CPC
Class: |
A61P 9/10 20180101; A61P
19/02 20180101; A61P 31/04 20180101; C07D 495/04 20130101; A61P
17/00 20180101; A61P 1/04 20180101; A61P 35/00 20180101; A61P 11/06
20180101; A61P 11/00 20180101; A61P 29/00 20180101; A61P 3/10
20180101; C07D 498/02 20130101; A61P 43/00 20180101 |
Class at
Publication: |
514/253.04 ;
544/362 |
International
Class: |
A61K 31/496 20060101
A61K031/496; C07D 498/02 20060101 C07D498/02 |
Claims
1. A compound having a structure selected from the group consisting
of structure (I), structure (II), and structure (III): ##STR180##
or a stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof, wherein: R.sub.1 is selected from the group
consisting of hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), and --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one substituent selected
from the group consisting of halogen, keto, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, and di-(C.sub.1-6
alkyl)amino; R.sub.2 is selected from the group consisting of
-C(.dbd.O)-(C.sub.1-6 alkyl), --NO, --NO.sub.2, --CONH.sub.2,
--C(.dbd.O)--NH(C.sub.1-6 alkyl), --C(.dbd.O)--N(C.sub.1-6
alkyl).sub.2, --C(.dbd.O)--NH--(5-7 membered heterocycle),
--C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3, --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), and --OC(.dbd.O)-(C.sub.1-6 alkyl);
R.sub.3 is selected from the group consisting of C.sub.1-8 alkyl,
--(CH.sub.2).sub.y-(C.sub.6-18 aryl), and --(CH.sub.2).sub.y-(5-7
membered heterocycle), wherein y is 0 to 4, and wherein R.sub.3 is
unsubstituted or substituted with at least one substituent selected
from the group consisting of halogen, hydroxy,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl), keto, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, and di-(C.sub.1-6
alkyl)amino; X is selected from the group consisting of hydrogen,
halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino,
and di-(C.sub.1-6 alkyl)amino; Y is selected from the group
consisting of hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy,
C.sub.1-6 alkylamino, and di-(C.sub.1-6 alkyl)amino; and n is 0, 1,
or 2.
2. The compound of claim 1, wherein R.sub.3 is selected from the
group consisting of thiophenyl, furanyl, 4-hydroxyphenyl,
4-methoxyphenyl, and 5-F-thiophenyl.
3. The compound of claim 1, wherein R.sub.2 is selected from the
group consisting of --C(.dbd.O)OCH.sub.2CH.sub.3,
--C(.dbd.O)OCH(CH.sub.3).sub.2, and --CN.
4. The compound of claim 1, wherein X is hydrogen.
5. The compound of claim 1, wherein Y is hydrogen.
6. The compound of claim 1, wherein n is 1.
7. The compound of claim 1, having a structure: ##STR181## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
8. The compound of claim 1, having a structure: ##STR182## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
9. The compound of claim 1, having a structure: ##STR183## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
10. The compound of claim 1, having a structure: ##STR184## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
11. The compound of claim 1, having a structure: ##STR185## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof
12. The compound of claim 1, having a structure: ##STR186## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
13. The compound of claim 1, having a structure: ##STR187## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
14. The compound of claim 1, having a structure: ##STR188## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
15. The compound of claim 1, having a structure: ##STR189## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
16. The compound of claim 1, having a structure: ##STR190## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
17. The compound of claim 1, having a structure: ##STR191## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
18. The compound of claim 1, having a structure: ##STR192## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
19. The compound of claim 1, having a structure: ##STR193## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
20. The compound of claim 1, having a structure: ##STR194## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
21. The compound of claim 1, having a structure: ##STR195## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
22. The compound of claim 1, having a structure: ##STR196## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
23. The compound of claim 1, having a structure: ##STR197## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
24. The compound of claim 1, having a structure: ##STR198## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
25. The compound of claim 1, having a structure: ##STR199## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
26. The compound of claim 1, having a structure: ##STR200## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
27. The compound of claim 1, having a structure: ##STR201## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
28. The compound of claim 1, having a structure: ##STR202## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
29. The compound of claim 1, having a structure: ##STR203## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
30. The compound of claim 1, having a structure: ##STR204## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
31. The compound of claim 1, having a structure: ##STR205## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
32. The compound of claim 1, having a structure: ##STR206## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
33. The compound of claim 1, having a structure: ##STR207## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof.
34. A pharmaceutical composition for treating a disease or disorder
wherein macrophage migration inhibitory factor is pathogenic, the
pharmaceutical composition comprising a compound of claim 1 in
combination with a pharmaceutically acceptable carrier or
diluent.
35. The pharmaceutical composition of claim 34, further comprising
a drug for treating the disease or disorder, wherein the drug has
no measurable macrophage migration inhibitory factor inhibiting
activity.
36. The pharmaceutical composition of claim 34, further comprising
a drug selected from the group consisting of a nonsteroidal
anti-inflammatory drug, an anti-infective drug, a beta stimulant, a
steroid, an antihistamine, an anticancer drug, an asthma drug, a
sepsis drug, an arthritis drug, and an immunosuppressive drug.
37. A method for treating a disease or disorder wherein macrophage
migration inhibitory factor is pathogenic, comprising administering
to a patient in need thereof an effective amount of a compound
having a structure selected from the group consisting of structure
(I), structure (II), and structure (III): ##STR208## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof, wherein: R.sub.1 is selected from the group
consisting of hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), and --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one substituent selected
from the group consisting of halogen, keto, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, and di-(C.sub.1-6
alkyl)amino; R.sub.2 is selected from the group consisting of
--C(.dbd.O)-(C.sub.1-6 alkyl), --NO, --NO.sub.2, --CONH.sub.2,
--C(.dbd.O)--NH(C.sub.1-6 alkyl), --C(.dbd.O)--N(C.sub.1-6
alkyl).sub.2, --C(.dbd.O)--NH-(5-7 membered heterocycle),
--C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3, --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), and --OC(.dbd.O)-(C.sub.1-6 alkyl);
R.sub.3 is selected from the group consisting of C.sub.1-8 alkyl,
--(CH.sub.2).sub.y-(C.sub.6-18 aryl), and --(CH.sub.2).sub.y-(5-7
membered heterocycle), wherein y is 0 to 4, and wherein R.sub.3 is
unsubstituted or substituted with at least one substituent selected
from the group consisting of halogen, hydroxy,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl), keto, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, and di-(C.sub.1-6
alkyl)amino; X is selected from the group consisting of hydrogen,
halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino,
and di-(C.sub.1-6 alkyl)amino; Y is selected from the group
consisting of hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy,
C.sub.1-6 alkylamino, and di-(C.sub.1-6 alkyl)amino; and n is 0, 1,
or 2; whereby macrophage migration inhibitory factor is
inhibited.
38. The method of claim 37, wherein the disease or disorder is
selected from the group consisting of inflammation, septic shock,
arthritis, cancer, acute respiratory distress syndrom, an
inflammatory disease, rheumatoid arthritis, osteoarthritis,
inflammatory bowel disease, asthma, an autoimmune disorder,
diabetes, asthma, multiple sclerosis, Cushing's disease,
cardiovascular disease, restenosis, and atherosclerosis.
Description
RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Application No. 60/665,236, filed Mar. 24, 2005, and U.S.
Provisional Application No. 60/733,657, filed Nov. 4, 2005. All
above-referenced prior applications are incorporated by reference
herein in their entirety and are hereby made a portion of this
specification.
FIELD OF THE INVENTION
[0002] The present invention relates to organic compounds, e.g.,
thienopyridinone derivatives, that are macrophage migration
inhibitory factor (MIF) inhibitors.
BACKGROUND OF THE INVENTION
[0003] The lymphokine, macrophage migration inhibitory factor
(MIF), has been identified as a mediator of the function of
macrophages in host defense and its expression correlates with
delayed hypersensitivity, immunoregulation, inflammation, and
cellular immunity. Although MIF was first characterized as being
able to block macrophage migration, MIF also appears to effect
macrophage adherence; induce macrophage to express
interleukin-1-beta, interleukin-6, and tumor necrosis factor alpha;
up-regulate HLA-DR; increase nitric oxide synthase and nitric oxide
concentrations; and activate macrophage to kill Leishmania donovani
tumor cells and inhibit Mycoplasma avium growth, by a mechanism
different from that effected by interferon-gamma.
[0004] In addition to its potential role as an immunoevasive
molecule, MIF can act as an immunoadjuvant when given with bovine
serum albumin or HIV gp120 in incomplete Freunds or liposomes,
eliciting antigen induced proliferation comparable to that of
complete Freunds. Also, MIF has been described as a glucocorticoid
counter regulator and angiogenic factor. As one of the few proteins
that is induced and not inhibited by glucocorticoids, it serves to
attenuate the immunosuppressive effects of glucocorticoids. As
such, it is viewed as a powerful element that regulates the
immunosuppressive effects of glucocorticoids. Hence, when its
activities/gene expression are overinduced by the administration of
excess exogenous glucocorticoids (for example when clinical
indicated to suppress inflammation, immunity and the like), there
is significant toxicity because MIF itself exacerbates the
inflammatory/immune response. See Bucala et al., Ann. Rep. Med.
Chem. 33:243-252, 1998.
[0005] The interest in developing MIF inhibitors derives from the
observation that MIF is known for its cytokine activity
concentrating macrophages at sites of infection, and cell-mediated
immunity. Moreover, MIF is known as a mediator of macrophage
adherence, phagocytosis, and tumoricidal activity. Hence, the
inhibition of MIF results in the indirect inhibition of cytokines,
growth factors, chemokines, and lymphokines that the macrophage may
otherwise bring to a site of inflammation.
SUMMARY OF THE INVENTION
[0006] As MIF has been identified in a variety of tissues and has
been associated with numerous pathological events, there exists a
need for pharmaceutical compositions comprising MIF inhibitors, as
well as methods relating to the use thereof to treat, for example,
immune related disorders or other MIF induced pathological events,
such as tumor associated angiogenesis. The preferred embodiments
fulfill these needs, and provide other advantages as well.
[0007] In preferred embodiments, MIF inhibitors are provided that
have the following general structures (I), (II), or (III),
including forms such as stereoisomers, free forms, pharmaceutically
acceptable salts or esters thereof, solvates, or combinations of
such forms, wherein n, R.sub.1, R.sub.2, R.sub.3, X, and Y are as
defined below: ##STR2##
[0008] The MIF inhibitors of preferred embodiments have utility
over a wide range of therapeutic applications, and may be employed
to treat a variety of disorders, illnesses, or pathological
conditions including, but not limited to, a variety of immune
related responses, tumor growth (e.g., prostate cancer, etc.),
glomerulonephritis, inflammation, malarial anemia, septic shock,
tumor associated angiogenesis, vitreoretinopathy, psoriasis, graft
versus host disease (tissue rejection), atopic dermatitis,
rheumatoid arthritis, inflammatory bowel disease, otitis media,
Crohn's disease, acute respiratory distress syndrome, delayed-type
hypersensitivity, and others.
[0009] Therapeutic methods include administering an effective
amount of one or more MIF inhibitors as provided by the preferred
embodiments, preferably in the form of a pharmaceutical
composition, to patient in need thereof. Pharmaceutical
compositions are provided containing one or more MIF inhibitors of
preferred embodiments in combination with pharmaceutically
acceptable carrier(s) and/or diluent(s).
[0010] Accordingly, in a first aspect a compound is provided having
structure (I), structure (II), or structure (III): ##STR3## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof, wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x--C.sub.6-18 aryl), or --CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; R.sub.2 is --NO, --NO.sub.2,
--CONH.sub.2, --C(.gradient.O--NH(C.sub.1-6 alkyl),
--C(.dbd.O)--N(C.sub.1-6 alkyl).sub.2, --C(.dbd.O)--NH-5-7 membered
heterocycle), --C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O--C.sub.1-6
alkyl), or --OC(.dbd.O)C.sub.1-6 alkyl); R.sub.3 is C.sub.1-8
alkyl, --(CH.sub.2).sub.y--C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; X is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; Y is hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6
alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6 alkyl)amino; and n
is 0, 1, or 2.
[0011] In an embodiment of the first aspect, R.sub.3 is thiophenyl,
furanyl, 4-hydroxyphenyl, 4-methoxyphenyl, or 5-F-thiophenyl. In an
embodiment of the first aspect, R.sub.2 is --CN,
--C(.dbd.O)OCH.sub.2CH.sub.3, or --C(.dbd.O)OCH(CH.sub.3).sub.2. In
an embodiment of the first aspect, X is hydrogen, and/or Y is
hydrogen. In an embodiment of the first aspect, n is 1. In an
embodiment of the first aspect, the compound is in a form of a
salt. In an embodiment of the first aspect, the compound is for use
as a pharmaceutical composition.
[0012] In an embodiment of the first aspect, a pharmaceutical
composition is provided comprising a compound of the first aspect
in association with at least one pharmaceutically acceptable
excipient. In an embodiment of the first aspect, a pharmaceutical
composition is provided comprising a compound of the first aspect
and at least one additional pharmaceutically active agent.
[0013] In an embodiment of the first aspect, use of a compound of
the first aspect for the manufacture of a medicament for the
treatment of a disease or disorder mediated by macrophage migration
inhibitory factor is provided.
[0014] In an embodiment of the first aspect, a method is provided
for treating a disease or disorder such as inflammation, septic
shock, arthritis, cancer, acute respiratory distress syndrome,
inflammatory disease, rheumatoid arthritis, osteoarthritis,
inflammatory bowel disease, asthma, autoimmune disorder, Lyme
disease, Lupus, Acquired Immune Deficiency Syndrome, diabetes,
multiple sclerosis, congestive heart failure, cardiovascular
disease restenosis, and atherosclerosis, the method comprising
administering to a patient in need thereof an effective amount of
the compound of the first aspect. The method can also comprise
administering a compound of the first aspect in combination with
another pharmaceutically active agent, either simultaneously or in
sequence.
[0015] In a second aspect, a pharmaceutical composition is provided
comprising a compound having structure (I), structure (II), or
structure (III): ##STR4## or a stereoisomer, or a pharmaceutically
acceptable salt, ester, or solvate thereof, wherein R.sub.1 is
hydrogen, C.sub.1-8 alkyl, --(CH.sub.2).sub.x-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.x-(5-7 membered heterocycle), wherein x is 0 to 4,
and wherein R.sub.1 is unsubstituted or substituted with at least
one of halogen, keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; R.sub.2 is --NO,
--NO.sub.2, --CONH.sub.2, --C(.dbd.O)--NH(C.sub.1-6 alkyl),
--C(.dbd.O)--N(C.sub.1-6 alkyl).sub.2, --C(.dbd.O)--NH-(5-7
membered heterocycle), --C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O)-(C.sub.1-6 alkyl); R.sub.3 is C.sub.1-8
alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; X is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; Y is hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6
alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6 alkyl)amino; and n
is 0, 1, or 2; in combination with a pharmaceutically acceptable
carrier or diluent.
[0016] In a third aspect, a method for reducing macrophage
migration inhibitory factor activity in a patient in need thereof
is provided, comprising administering to the patient an effective
amount of a compound having the structure: ##STR5## or a
stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof, wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; R.sub.2 is --NO, --NO.sub.2,
--CONH.sub.2, --C(.dbd.O)--NH(C.sub.1-6 alkyl),
--C(.dbd.O)--N(C.sub.1-6 alkyl).sub.2, --C(.dbd.O)--NH-5-7 membered
heterocycle), --C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-C.sub.1-6 alkyl),
or --OC(.dbd.O)-(C.sub.1-6 alkyl); R.sub.3 is C.sub.1-8 alkyl,
--(CH.sub.2).sub.y-(C.sub.6-18 aryl), or --(CH.sub.2).sub.y-(5-7
membered heterocycle), wherein y is 0 to 4, and wherein R.sub.3 is
unsubstituted or substituted with at least one of halogen, hydroxy,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl), keto, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; X is hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6
alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6 alkyl)amino; Y is
hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; and n is 0, 1, or 2.
[0017] In a fourth aspect, a method for treating a disease or a
disorder in an animal is provided, the method comprising
administering to the animal an effective amount of a compound
having the structure: ##STR6## or a stereoisomer, or a
pharmaceutically acceptable salt, ester, or solvate thereof,
wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; R.sub.2 is --NO, --NO.sub.2,
--CONH.sub.2, --C(.dbd.O)--NH(C.sub.1-6 alkyl),
--C(.dbd.O)--N(C.sub.1-6 alkyl).sub.2, --C(.dbd.O)--NH-(5-7
membered heterocycle), --C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or -OC(.dbd.OHC.sub.1-6 alkyl); R.sub.3 is C.sub.1-8 alkyl,
--(CH.sub.2).sub.y-(C.sub.6-18 aryl), or --(CH.sub.2).sub.y-(5-7
membered heterocycle), wherein y is 0 to 4, and wherein R.sub.3 is
unsubstituted or substituted with at least one of halogen, hydroxy,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl), keto, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; X is hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6
alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6 alkyl)amino; Y is
hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; and n is 0, 1, or 2.
[0018] In an embodiment of the fourth aspect, the disease or
disorder is inflammation, septic shock, arthritis, cancer, acute
respiratory distress syndrome, an inflammatory disease, rheumatoid
arthritis, osteoarthritis, inflammatory bowel disease, asthma, an
autoimmune disorder, diabetes, or multiple sclerosis.
[0019] In an embodiment of the fourth aspect, an immune response is
suppressed.
[0020] In an embodiment of the fourth aspect, angiogenesis is
decreased.
[0021] In an embodiment of the fourth aspect, the disease is
associated with excess glucocorticoid levels, for example,
Cushing's disease.
[0022] In a fifth aspect, a pharmaceutical composition is provided
for treating a disease or disorder wherein macrophage migration
inhibitory factor is pathogenic, the pharmaceutical composition
comprising a compound of structure (I), (II), or (III): ##STR7## or
a stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof, wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; R.sub.2 is --NO, --NO.sub.2,
--CONH.sub.2, --C(.dbd.O)--NH(C.sub.1-6 alkyl),
--C(.dbd.O)--N(C.sub.1-6 alkyl).sub.2, --C(.dbd.O)--NH-(5-7
membered heterocycle), --C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O)-(C.sub.1-6 alkyl); R.sub.3 is C.sub.1-8
alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; X is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; Y is hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6
alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6 alkyl)amino; and n
is 0, 1, or 2.
[0023] In a sixth aspect, a pharmaceutical composition is provided
for treating a disease or disorder wherein macrophage migration
inhibitory factor is pathogenic, the pharmaceutical composition
comprising a compound of structure (I), (II), or (III) in
combination with a drug for treating the disease or disorder,
wherein the drug has no measurable MIF inhibiting activity, and
wherein structures (I), (II), and (III) are as follows: ##STR8## or
a stereoisomer, or a pharmaceutically acceptable salt, ester, or
solvate thereof, wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; R.sub.2 is --NO, --NO.sub.2,
--CONH.sub.2, --C(.dbd.O)--NH(C.sub.1-6 alkyl),
--C(.dbd.O)--N(C.sub.1-6 alkyl).sub.2, --C(.dbd.O)NH-5-7 membered
heterocycle), --C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or -OC(.dbd.O)-C.sub.1-6 alkyl); R.sub.3 is C.sub.1-8
alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; X is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; Y is hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6
alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6alkyl)amino; and n is
0, 1, or 2.
[0024] In an embodiment of the sixth aspect, the disease or
disorder is inflammation, septic shock, or rheumatoid arthritis and
the drug is a steroid.
[0025] In an embodiment of the sixth aspect, the disease or
disorder is asthma or acute respiratory distress, and the drug is a
corticosteroid, for example, cortisone, hydrocortisone,
methylprednisolone, prednisone, prednisolone, betamethesone,
beclomethasone dipropionate, budesonide, dexamethasone sodium
phosphate, flunisolide, fluticasone propionate, triamcinolone
acetonide, betamethasone, fluocinolone, fluocinonide, betamethasone
dipropionate, betamethasone valerate, desonide, desoximetasone,
fluocinolone, triamcinolone, triamcinolone acetonide, clobetasol
propionate, or dexamethasone.
[0026] In an embodiment of the sixth aspect, the disease or
disorder is asthma or acute respiratory distress, and the drug is
beclomethasone, fluticasone, triamcinolone, mometasone, prednisone,
prednisolone, methylprednisolone, an azatadine,
carbinoxamine/pseudoephedrine, cetirizine, cyproheptadine,
dexchlorphenirarnine, fexofenadine, loratadine, promethazine,
tripelennamine, brompheniramine, cholopheniramine, clemastine,
diphenhydramine, or epinephrine.
[0027] In an embodiment of the sixth aspect, the disease or
disorder is irritable bowel disease, and the drug is azathioprine
or a corticosteroid.
[0028] In an embodiment of the sixth aspect, the disease or
disorder is cancer, and the drug is paclitaxel.
[0029] In an embodiment of the sixth aspect, the disease or
disorder is an immune disorder and the drug is an immunosuppressive
compound. The immune disorder can be Lyme disease, Lupus, or
Acquired Immune Deficiency Syndrome. The drug can be a protease
inhibitor, a nucleoside reverse transcriptase inhibitor, a
nucleotide reverse transcriptase inhibitor, a non-nucleoside
reverse transcriptase inhibitor, a biological response modifier, a
compound that inhibits or interferes with tumor necrosing factor,
or an antiviral. Examples of drugs include indinavir, amprenavir,
saquinavir, lopinavir, ritonavir, nelfinavir zidovudine, abacavir,
lamivudine, idanosine, zalcitabine, stavudine, tenofovir disoproxil
fumarate delavirdine, efavirenz, nevirapine, etanercept,
infliximab, amivudine, or zidovudine.
[0030] In a seventh aspect, a pharmaceutical composition is
provided for treating a disease or disorder wherein macrophage
migration inhibitory factor is pathogenic, the pharmaceutical
composition comprising a compound of structure (I), (II), or (III)
and a drug such as a nonsteroidal anti-inflammatory drug, an
anti-infective drug, a beta stimulant, a steroid, an antihistamine,
an anticancer drug, an asthma drug, a sepsis drug, an arthritis
drug, or an immunosuppressive drug, and wherein structures (I),
(II), and (III) are as follows: ##STR9## or a stereoisomer, or a
pharmaceutically acceptable salt, ester, or solvate thereof,
wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; R.sub.2 is --NO, --NO.sub.2,
--CONH.sub.2, --C(.dbd.O)--NH(C.sub.1-6 alkyl),
--C(.dbd.O)--N(C.sub.1-6 alkyl.sub.2, --C(.dbd.O)--NH-(5-7 membered
heterocycle), --C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O)-(C.sub.1-6 alkyl); R.sub.3 is C.sub.1-8
alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; X is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; Y is hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6
alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6 alkyl)amino; and n
is 0, 1, or 2. The beta stimulant can be a bronchodilator, an
inhalation corticosteroid, or a hormone. The inhalation
corticosteroid can be beclomethasone, fluticasone, triamcinolone,
mometasone, prednisone, prednisolone, or methylprednisolone. The
antihistamine can be azatadine, carbinoxamine/pseudoephedrine,
cetirizine, cyproheptadine, dexchlorpheniramine, fexofenadine,
loratadine, promethazine, tripelennamine, brompheniramine,
cholopheniramine, clemastine, diphenhydramine, or epinephrine. The
steroid can be cortisone, hydrocortisone, methylprednisolone,
prednisone, prednisolone, betamethesone, beclomethasone
dipropionate, budesonide, dexamethasone sodium phosphate,
flunisolide, fluticasone propionate, triamcinolone acetonide,
betamethasone, fluocinolone, fluocinonide, betamethasone
dipropionate, betamethasone valerate, desonide, desoximetasone,
fluocinolone, triamcinolone, triamcinolone acetonide, clobetasol
propionate, or dexamethasone. The anti-infective drug can be an
anthelmintic, an aminoclycoside, an antifungal antibiotic, a
cephalosporin, a beta-lactam antibiotic, chloramphenicol, a
macrolide, a penicillin, a tetracycline, bacitracin, clindamycin,
colistimethate sodium, polymyxin b sulfate, vancomycin, antivirals,
acyclovir, amantadine, didanosine, efavirenz, foscamet,
ganciclovir, indinavir, lamivudine, nelfinavir, ritonavir,
saquinavir, stavudine, valacyclovir, valganciclovir, zidovudine, a
quinolone, a sulfonamide, furazolidone, metronidazole, pentamidine,
sulfanilamidum crystallinum, gatifloxacin,
sulfamethoxazole/trimethoprim, mebendazole, gentamicin, neomycin,
tobramycin, amphotericin b, fluconazole, griseofulvin,
itraconazole, ketoconazole, nystatin, micatin, tolnaftate,
cefaclor, cefazolin, cefotaxime, ceftazidime, ceftriaxone,
cefuroxime, cephalexin, cefotetan, meropenem, azithromycin,
clarithromycin, erythromycin, penicillin G sodium salt,
amoxicillin, ampicillin, dicloxacillin, nafcillin, piperacillin,
ticarcillin, doxycycline, minocycline, tetracycline, ciprofloxacin,
levofloxacin, sulfadiazine, sulfisoxazole, or dapsone. The
nonsteroidal anti-inflammatory drug can be celecoxib, rofecoxib,
aspirin, celecoxib, choline magnesium trisalicylate, diclofenac
potassium, diclofenac sodium, diflunisal, etodolac, fenoprofen,
flurbiprofen, ibuprofen, indomethacin, ketoprofen, ketorolac,
melenamic acid, nabumetone, naproxen, naproxen sodium, oxaprozin,
piroxicam, rofecoxib, salsalate, sulindac, or tolmetin.
[0031] In an eighth aspect, a process for preparing a compound of
Formula (I-7) for use as a macrophage migration inhibitory factor
inhibitor is provided, the process comprising the steps of reacting
POCl.sub.3 with a compound of Formula (I-3) ##STR10## wherein
R.sub.2 is --NO, --NO.sub.2, --CONH.sub.2,
--C(.dbd.O)--NH(C.sub.1-6 alkyl), --C(.dbd.O)--N(C.sub.1-6
alkyl).sub.2, --C(.dbd.O)--NH-(5-7 membered heterocycle),
--C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O)-(C.sub.1-6 alkyl); wherein X is hydrogen,
halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino,
or di-(C.sub.1-6 alkyl)amino; and wherein Y is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; thereby yielding a compound of Formula
(I-4): ##STR11## reacting the compound of Formula (I-4) with
NH.sub.4OAc, thereby yielding a compound of Formula (I-5):
##STR12## reacting the compound of Formula (I-5) with a compound of
Formula (I-6): ##STR13## wherein R.sub.3 is C.sub.1-8 alkyl,
--(CH.sub.2).sub.y-(C.sub.6-18 aryl), or --(CH.sub.2).sub.y-(5-7
membered heterocycle), wherein y is 0 to 4, and wherein R.sub.3 is
unsubstituted or substituted with at least one of halogen, hydroxy,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl), keto, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; thereby yielding a compound of Formula (I-7):
##STR14## wherein the compound of Formula (I-7) is suitable for use
as a macrophage migration inhibitory factor inhibitor.
[0032] In a ninth aspect, a process for preparing a compound of
Formula (I-8) suitable for use as a macrophage migration inhibitory
factor inhibitor is provided, the process comprising the steps of
reacting a compound of Formula (I-7): ##STR15## wherein R.sub.2 is
--NO, --NO.sub.2, --CONH.sub.2, --C(.dbd.O)--N(C.sub.1-6 alkyl),
--C(.dbd.O)--N(C.sub.1-6 alkyl).sub.2, --C(.dbd.O)--NH-(5-7
membered heterocycle), --C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O-(C.sub.1-6 alkyl); wherein R.sub.3 is
C.sub.1-8 alkyl, --(CH.sub.2).sub.x-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; wherein X is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; and wherein Y is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; thereby yielding a compound of Formula
(I-8): ##STR16## wherein the compound of Formula (I-8) is suitable
for use as a macrophage migration inhibitory factor inhibitor.
[0033] In an embodiment of the ninth aspect, R.sub.1 is
##STR17##
[0034] In an embodiment of the ninth aspect, R.sub.2 is
--C(.dbd.O)OCH.sub.2CH.sub.3 or --CN.
[0035] In an embodiment of the ninth aspect, R.sub.3 ##STR18##
[0036] In a tenth aspect, process is provided for preparing a
compound of Formula (I-7) suitable for use as a macrophage
migration inhibitory factor inhibitor, the process comprising the
steps of reacting a compound of Formula (I-5): ##STR19## wherein
R.sub.2 is --NO, --NO.sub.2, --CONH.sub.2,
--C(.dbd.O)--NH(C.sub.1-6 alkyl), --C(.dbd.O)--N(C.sub.1-6
alkyl).sub.2, --C(.dbd.O)--NH-(5-7 membered heterocycle),
--C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O)-(C.sub.1-6 alkyl); wherein X is hydrogen,
halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino,
or di-(C.sub.1-6 alkyl)amino; and wherein Y is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; with a compound of Formula (I-9):
##STR20## wherein boc is t-butyloxycarbonyl, thereby yielding a
compound of Formula (I-10): ##STR21## reacting the compound of
Formula (I-10) with a compound having the formula R.sub.1-Z,
wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-.sub.18 aryl), or
--(CH.sub.2).sub.x-(5-7 membered heterocycle), wherein x is 0 to 4,
and wherein R.sub.1 is unsubstituted or substituted with at least
one of halogen, keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; and wherein Z is Cl, Br,
I, or B(OH).sub.2, thereby yielding a compound of Formula (I-11):
##STR22## reacting the compound of Formula (I-11) with
trifluoroacetic acid to yield a compound of Formula (I-1 2):
##STR23## reacting the compound of Formula (I-12) with
R.sub.3--C(.dbd.O)-Z, wherein Z is Cl, Br, or I, or wherein R.sub.3
is C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; whereby a compound of Formula (I-8) is
obtained: ##STR24## wherein the compound of Formula (I-8) is
suitable for use as a macrophage migration inhibitory factor
inhibitor.
[0037] In an eleventh aspect, a process is provided for preparing a
compound of Formula (I-7a) suitable for use as a macrophage
migration inhibitory factor inhibitor, the process comprising the
steps of reacting methylcyanoacetate with a compound of Formula
(I-1 3): ##STR25## wherein X is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; and wherein Y is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; to yield a compound of Formula (I-1 4): ##STR26##
reacting NaOEt with the compound of Formula (I-14) to yield a
compound of Formula (I-3a): ##STR27## reacting the compound of
Formula (I-3a) with POCl.sub.3, thereby yielding a compound of
Formula (I-5a): ##STR28## reacting the compound of Formula (I-5a)
with a compound of Formula (I-6): ##STR29## wherein R.sub.3 is
C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle) wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino, thereby yielding a compound of Formula
(I-7a): ##STR30## wherein the compound of Formula (I-7a) is
suitable for use as a macrophage migration inhibitory factor
inhibitor.
[0038] In a twelfth aspect, a process is provided for preparing a
compound of Formula (I-8a) suitable for use as a macrophage
migration inhibitory factor inhibitor, the process comprising the
steps of reacting a compound of Formula (I-7a): ##STR31## wherein
R.sub.3 is C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl),
or --(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to
4, and wherein R.sub.3 is unsubstituted or substituted with at
least one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; with a compound having the formula
R.sub.1-Z, wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-.sub.18 aryl), or
--(CH.sub.2).sub.x-(5-7 membered heterocycle), wherein x is 0 to 4,
and wherein R.sub.1 is unsubstituted or substituted with at least
one of halogen, keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; thereby yielding a
compound of Formula (I-8a): ##STR32## wherein the compound of
Formula (8a) is suitable for use as a macrophage migration
inhibitory factor inhibitor.
[0039] In an embodiment of the twelfth aspect, R.sub.1 is
##STR33##
[0040] In an embodiment of the twelfth aspect, R.sub.3 is
##STR34##
[0041] In a thirteenth aspect, a process is provided for preparing
a compound of Formula (I-8a) suitable for use as a macrophage
migration inhibitory factor inhibitor, the process comprising the
steps of reacting a compound of Formula (I-5a): ##STR35## wherein X
is hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; and wherein Y is
hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; with a compound of
Formula (I-9): ##STR36## wherein boc is t-butyloxycarbonyl, thereby
yielding a compound of Formula (I-10): ##STR37## reacting the
compound of Formula (I-10) with a compound having the formula
R.sub.1-Z, wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; and wherein Z is Cl, Br, I, or
B(OH).sub.2, thereby yielding a compound of Formula (I-11a):
##STR38## reacting the compound of Formula (I-11a) with
trifluoroacetic acid to yield a compound of Formula (I-12a):
##STR39## reacting the compound of Formula (I-12a) with
R.sub.3-C(.dbd.O)-Z, wherein Z is Cl, Br, or I, and wherein R.sub.3
is C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; whereby a compound of Formula (I-8a) is
obtained: ##STR40## wherein the compound of Formula (I-8) is
suitable for use as a macrophage migration inhibitory factor
inhibitor.
[0042] A process for preparing a compound of Formula (II-7)
suitable for use as a macrophage migration inhibitory factor
inhibitor, the process comprising the steps of reacting POCl.sub.3
with a compound of Formula (II-3): ##STR41## wherein R.sub.2 is
--NO, --NO.sub.2, --CONH.sub.2, --C(.dbd.O)--NH(C.sub.1-6 alkyl),
--C(.dbd.O)--N(C.sub.1-6 alkyl).sub.2, --C(.dbd.O)--NH-(5-7
membered heterocycle), --C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O)-(C.sub.1-6 alkyl); wherein X is hydrogen,
halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino,
or di-(C.sub.1-6 alkyl)amino; and wherein Y is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; thereby yielding a compound of Formula
(II-4): ##STR42## reacting the compound of Formula (II-4) with
NH.sub.4OAc, thereby yielding a compound of Formula (II-5):
##STR43## reacting the compound of Formula (II-5) with a compound
of Formula (I-6): ##STR44## wherein R.sub.3 is C.sub.1-8 alkyl,
--(CH.sub.2).sub.y-(C.sub.6-18 aryl), or --(CH.sub.2).sub.y-(5-7
membered heterocycle), wherein y is 0 to 4, and wherein R.sub.3 is
unsubstituted or substituted with at least one of halogen, hydroxy,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl), keto, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; thereby yielding a compound of Formula (II-7):
##STR45## wherein the compound of Formula (II-7) is suitable for
use as a macrophage migration inhibitory factor inhibitor.
[0043] In a fourteenth aspect, a process is provided for preparing
a compound of Formula (II-8) suitable for use as a macrophage
migration inhibitory factor inhibitor, the process comprising the
steps of reacting a compound of Formula (II-7): ##STR46## wherein
R.sub.2 is --NO, --NO.sub.2, --CONH.sub.2,
--C(.dbd.O)--NH(C.sub.1-6 alkyl), --C(.dbd.O)--N(C.sub.1-6 alkyl)
.sub.2, --C(.dbd.O)--NH-(5-7 membered heterocycle),
--C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O)-(C.sub.1-6 alkyl); wherein R.sub.3 is
C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-8 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; wherein X is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; and wherein Y is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; with a compound having the formula
R.sub.1-Z, wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino, and wherein Z is Cl, Br, I, or
B(OH).sub.2, thereby yielding a compound of Formula (II-8):
##STR47## wherein the compound of Formula (II-8) is suitable for
use as a macrophage migration inhibitory factor inhibitor.
[0044] In a fifteenth aspect, a process is provided for preparing a
compound of Formula (II-7) suitable for use as a macrophage
migration inhibitory factor inhibitor, the process comprising the
steps of reacting a compound of Formula (II-5): ##STR48## wherein
R.sub.2 is --NO, --NO.sub.2, --CONH.sub.2,
--C(.dbd.O)--NH(C.sub.1-6 alkyl), --C(.dbd.O)--N(C.sub.1-6 alkyl)
.sub.2, --C(.dbd.O)--NH-(5-7 membered heterocycle),
--C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O)-(C.sub.1-6 alkyl); wherein X is hydrogen,
halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino,
or di-(C.sub.1-6 alkyl)amino; and wherein Y is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; with a compound of Formula (I-9):
##STR49## wherein boc is t-butyloxycarbonyl, thereby yielding a
compound of Formula (II-10): ##STR50## reacting the compound of
Formula (II-10) with a compound having the formula R.sub.1-Z,
wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl, 13
(CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino, and wherein Z is Cl, Br, I, or
B(OH).sub.2, thereby yielding a compound of Formula (II-11):
##STR51## reacting the compound of Formula (II-11) with
trifluoroacetic acid to yield a compound of Formula (II-12):
##STR52## reacting the compound of Formula (II-12) with
R.sub.3--C(.dbd.O)-Z, wherein Z is Cl, Br, or I, or wherein R.sub.3
is C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; whereby a compound of Formula (II-8) is
obtained: ##STR53## wherein the compound of Formula (II-8) is
suitable for use as a macrophage migration inhibitory factor
inhibitor.
[0045] In a sixteenth aspect, a process is provided for preparing a
compound of Formula (II-7a) suitable for use as a macrophage
migration inhibitory factor inhibitor, the process comprising the
steps of reacting methylcyanoacetate with a compound of Formula
(II-13): ##STR54## wherein X is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; and wherein Y is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; to yield a compound of Formula (II-14): ##STR55##
reacting NaOEt with the compound of Formula (II-14) to yield a
compound of Formula (II-3a): ##STR56## reacting the compound of
Formula (II-3a) with POCl.sub.3, thereby yielding a compound of
Formula (II-5a): ##STR57## reacting the compound of Formula (II-5a)
with a compound of Formula (I-6): ##STR58## wherein R.sub.3 is
C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; thereby yielding a compound of Formula
(II-7a): ##STR59## wherein the compound of Formula (II-7a) is
suitable for use as a macrophage migration inhibitory factor
inhibitor.
[0046] In a seventeenth aspect, a process is provided for preparing
a compound of Formula (II-8a) suitable for use as a macrophage
migration inhibitory factor inhibitor, the process comprising the
steps of reacting a compound of Formula (II-7a): ##STR60## wherein
R.sub.3 is C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl),
or --(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to
4, and wherein R.sub.3 is unsubstituted or substituted with at
least one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; wherein X is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; and wherein Y is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; with a compound having the formula
R.sub.1-Z, wherein R.sub.1 is hydrogen, C.sub.1-8
alkyl,--(CH.sub.2).sub.x-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.x-(5-7 membered heterocycle), wherein x is 0 to 4,
and wherein R.sub.1 is unsubstituted or substituted with at least
one of halogen, keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; and Z is Cl, Br, I, or
B(OH).sub.2, thereby yielding a compound of Formula (II-8a):
##STR61## wherein the compound of Formula (II-8a) is suitable for
use as a macrophage migration inhibitory factor inhibitor.
[0047] In a eighteenth aspect, a process is provided for preparing
a compound of Formula (II-8a) suitable for use as a macrophage
migration inhibitory factor inhibitor, the process comprising the
steps of reacting a compound of Formula (II-5a): ##STR62## wherein
X is hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy,
C.sub.1-6 alkylamino, or di-(C.sub.1-6 alkyl)amino; and wherein Y
is hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; with a compound of
Formula (I-9): ##STR63## wherein boc is t-butyloxycarbonyl, thereby
yielding a compound of Formula (II-10): ##STR64## reacting the
compound of Formula (II-10) with a compound having the formula
R.sub.1-Z, wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; and wherein Z is Cl, Br, I, or
B(OH).sub.2, thereby yielding a compound of Formula (II-11a):
##STR65## reacting the compound of Formula (II-11a) with
trifluoroacetic acid to yield a compound of Formula (II-12a):
##STR66## reacting the compound of Formula (II-12a) with
R.sub.3--C(.dbd.O)-Z, wherein Z is Cl, Br, or I, or wherein R.sub.3
is C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino, whereby a compound of Formula (II-8a) is
obtained: ##STR67## wherein the compound of Formula (II-8) is
suitable for use as a macrophage migration inhibitory factor
inhibitor.
[0048] In a nineteenth aspect, a process is provided for preparing
a compound of Formula (III-7) suitable for use as a macrophage
migration inhibitory factor inhibitor, the process comprising the
steps of reacting POCl.sub.3 with a compound of Formula (III-3):
##STR68## wherein R.sub.2 is --NO, --NO.sub.2, --CONH.sub.2,
--C(.dbd.O)--NH(C.sub.1-6 alkyl), --C(.dbd.O)--N(C.sub.1-6
alkyl).sub.2, --C(.dbd.O)--NH-(5-7 membered heterocycle),
--C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O)-(C.sub.1-6 alkyl); wherein X is hydrogen,
halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino,
or di-(C.sub.1-6 alkyl)amino; and wherein Y is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; thereby yielding a compound of Formula
(II4): ##STR69## reacting the compound of Formula (III-4) with
NH.sub.4OAc, thereby yielding a compound of Formula (III-5):
##STR70## reacting the compound of Formula (III-5) with a compound
of Formula (I-6): ##STR71## wherein R.sub.3 is C.sub.1-8 alkyl,
--(CH.sub.2).sub.y-(C.sub.6-18 aryl), or --(CH.sub.2).sub.y-(5-7
membered heterocycle), wherein y is 0 to 4, and wherein R.sub.3 is
unsubstituted or substituted with at least one of halogen, hydroxy,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl), keto, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; thereby yielding a compound of Formula (III-7):
##STR72## wherein the compound of Formula (III-7) is suitable for
use as a macrophage migration inhibitory factor inhibitor.
[0049] In a twentieth aspect, a process is provided for preparing a
compound of Formula (III-8) suitable for use as a macrophage
migration inhibitory factor inhibitor, the process comprising the
steps of reacting a compound of Formula (III-7): ##STR73## wherein
R.sub.2 is --NO, --NO.sub.2, --CONH.sub.2,
--C(.dbd.O)--NH(C.sub.1-6 alkyl), --C(.dbd.O)--N(C.sub.1-6 alkyl)
.sub.2, --C(.dbd.O)--NH-(5-7 membered heterocycle),
--C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O)-(C.sub.1-6 alkyl); wherein R.sub.3 is
C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; wherein X is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; and wherein Y is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; with a compound having the formula
R.sub.1-Z, wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; and wherein Z is Cl, Br, I, or
B(OH).sub.2, thereby yielding a compound of Formula (III-8):
##STR74## wherein the compound of Formula (III-8) is suitable for
use as a macrophage migration inhibitory factor inhibitor.
[0050] In a twenty-first aspect, a process is provided for
preparing a compound of Formula (III-7) suitable for use as a
macrophage migration inhibitory factor inhibitor, the process
comprising the steps of reacting a compound of Formula (III-5):
##STR75## wherein R.sub.2 is --NO, --NO.sub.2, --CONH.sub.2,
--C(.dbd.O)--NH(C.sub.1-6 alkyl), --C(.dbd.O)--N(C.sub.1-6
alkyl).sub.2, --C(.dbd.O)--NH-(5-7 membered heterocycle),
--C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), or --OC(.dbd.O)-C.sub.1-6 alkyl); wherein X is hydrogen,
halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino,
or di-(C.sub.1-6 alkyl)amino; and wherein Y is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; with a compound of Formula (I-9):
##STR76## wherein boc is t-butyloxycarbonyl, thereby yielding a
compound of Formula (III-10): ##STR77## reacting the compound of
Formula (III-10) with a compound having the formula R.sub.1-Z,
wherein R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; and wherein Z is Cl, Br, I, or
B(OH).sub.2, thereby yielding a compound of Formula (III-11):
##STR78## reacting the compound of Formula (III-11) with
trifluoroacetic acid to yield a compound of Formula (III-12):
##STR79## reacting the compound of Formula (III-12) with
R.sub.3-C(.dbd.O)-Z, wherein Z is Cl, Br, or I, or wherein R.sub.3
is C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to 4,
and wherein R.sub.3 is unsubstituted or substituted with at least
one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; whereby a compound of Formula (III-8) is
obtained: ##STR80## wherein the compound of Formula (III-8) is
suitable for use as a macrophage migration inhibitory factor
inhibitor.
[0051] In a twenty-second aspect, a process is provided for
preparing a compound of Formula (III-7a) suitable for use as a
macrophage migration inhibitory factor inhibitor, the process
comprising the steps of: reacting methylcyanoacetate with a
compound of Formula (III-13): ##STR81## wherein X is hydrogen,
halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino,
or di-(C.sub.1-6 alkyl)amino; and wherein Y is hydrogen, halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; to yield a compound of Formula (III-14):
##STR82## reacting NaOEt with the compound of Formula (III-14) to
yield a compound of Formula (III-3a): ##STR83## reacting the
compound of Formula (III-3a) with POCl.sub.3, thereby yielding a
compound of Formula (III-5a): ##STR84## reacting the compound of
Formula (III-5a) with a compound of Formula (I-6): ##STR85##
wherein R.sub.3 is C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18
aryl), or --(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y
is 0 to 4, and wherein R.sub.3 is unsubstituted or substituted with
at least one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl),
--CN, --C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6
alkyl), keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; thereby yielding a
compound of Formula (III-7a): ##STR86## wherein the compound of
Formula (III-7a) is suitable for use as a macrophage migration
inhibitory factor inhibitor.
[0052] In a twenty-third aspect, a process is provided for
preparing a compound of Formula (III-8a) suitable for use as a
macrophage migration inhibitory factor inhibitor, the process
comprising the steps of reacting a compound of Formula (III-7a):
##STR87## wherein R.sub.3 is C.sub.1-8 alkyl,
--(CH.sub.2).sub.y-(C.sub.6-18 aryl), or --(CH.sub.2).sub.y-(5-7
membered heterocycle) wherein y is 0 to 4, and wherein R.sub.3 is
unsubstituted or substituted with at least one of halogen, hydroxy,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl), keto, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; wherein X is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; and wherein Y is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; with a compound having the formula R.sub.1-Z, wherein
R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4, and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; and Z is Cl, Br, I, or B(OH).sub.2,
thereby yielding a compound of Formula (III-8a): ##STR88## wherein
the compound of Formula (III-8a) is suitable for use as a
macrophage migration inhibitory factor inhibitor.
[0053] In a twenty-fourth aspect, a process is provided for
preparing a compound of Formula (III-8a) suitable for use as a
macrophage migration inhibitory factor inhibitor, the process
comprising the steps of reacting a compound of Formula (III-5a):
##STR89## wherein X is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; and wherein Y is hydrogen, halogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; with a compound of Formula (I-9): ##STR90## wherein
boc is t-butyloxycarbonyl, thereby yielding a compound of Formula
(III-10): ##STR91## reacting the compound of Formula (III-10) with
a compound having the formula R.sub.1-Z, wherein R.sub.1 is
hydrogen, C.sub.1-8 alkyl, --(CH.sub.2).sub.x-(C.sub.6-18 aryl), or
--(CH.sub.2).sub.x-(5-7 membered heterocycle), wherein x is 0 to 4,
and wherein R.sub.1 is unsubstituted or substituted with at least
one of halogen, keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; and wherein Z is Cl, Br,
I, or B(OH).sub.2, thereby yielding a compound of Formula
(III-11a): ##STR92## reacting the compound of Formula (III-11a)
with trifluoroacetic acid to yield a compound of Formula (III-12a):
##STR93## reacting the compound of Formula (III-12a) with
R.sub.3--C(.dbd.O)-Z, wherein Z is Cl, Br, or I, and wherein
R.sub.3 is C.sub.1-8 alkyl, --(CH.sub.2).sub.y-(C.sub.6-18 aryl),
or --(CH.sub.2).sub.y-(5-7 membered heterocycle), wherein y is 0 to
4, and wherein R.sub.3 is unsubstituted or substituted with at
least one of halogen, hydroxy, --C(.dbd.O)-(C.sub.1-6 alkyl), --CN,
--C(.dbd.O)O-(C.sub.1-6 alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl),
keto, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; whereby a compound of Formula (III-8a)
is obtained: ##STR94## wherein the compound of Formula (III-8) is
suitable for use as a macrophage migration inhibitory factor
inhibitor.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
[0054] The following description and examples illustrate a
preferred embodiment of the present invention in detail. Those of
skill in the art will recognize that there are numerous variations
and modifications of this invention that are encompassed by its
scope. Accordingly, the description of a preferred embodiment
should not be deemed to limit the scope of the present
invention.
[0055] As an aid to understanding the preferred embodiments,
certain definitions are provided herein.
[0056] The terms "macrophage migration inhibitory activity" "MIF
activity" as used herein is a broad term, and is to be given its
ordinary and customary meaning to a person of ordinary skill in the
art (and is not to be limited to a special or customized meaning),
and refers without limitation to an activity or effect mediated at
least in part by macrophage migration inhibitory factor.
Accordingly, MIF activity includes, but is not limited to,
inhibition of macrophage migration, tautomerase activity (e.g.,
using phenylpyruvate or dopachrome), endotoxin induced shock,
inflammation, glucocorticoid counter regulation, induction of
thymidine incorporation into 3T3 fibroblasts, induction of erk
phosphorylation and MAP kinase activity.
[0057] The term "inhibitor" as used herein is a broad term, and is
to be given its ordinary and customary meaning to a person of
ordinary skill in the art (and is not to be limited to a special or
customized meaning), and refers without limitation to a molecule
(e.g., natural or synthetic compound) that can alter the
conformation of MIF and/or compete with a monoclonal antibody to
MIF and decrease at least one activity of MIF or its export from a
cell as compared to activity or export in the absence of the
inhibitor. In other words, an "inhibitor" alters conformation
and/or activity and/or export if there is a statistically
significant change in the amount of MIF measured, MIF activity, or
in MIF protein detected extracellularly and/or intracellularly in
an assay performed with an inhibitor, compared to the assay
performed without the inhibitor.
[0058] In general, MIF inhibitors inhibit the physiological
function of MIF, and thus are useful in the treatment of diseases
where MIF may be pathogenic.
[0059] In certain of the preferred embodiments, MIF inhibitors are
provided that are thienopyridinone derivatives having the following
structures (I), (II), and (III): ##STR95## including forms such as
stereoisomers, free forms, pharmaceutically acceptable salts or
esters thereof, solvates, and combinations of such forms; wherein
R.sub.1 is hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.x-(C.sub.6-18 aryl), or --(CH.sub.2).sub.x-(5-7
membered heterocycle), wherein x is 0 to 4 and wherein R.sub.1 is
unsubstituted or substituted with at least one of halogen, keto,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or
di-(C.sub.1-6 alkyl)amino; R.sub.2 is --CN, --NO, --NO.sub.2,
--CONH.sub.2, --C(.dbd.O)--NH(C.sub.1-6 alkyl),
--C(.dbd.O)--N(C.sub.1-6 alkyl.sub.2, --C(.dbd.O)--NH-(5-7 membered
heterocycle), --C(.dbd.O)-(5-7 membered heterocycle),
--C(.dbd.O)--N[(CH.sub.2).sub.2].sub.2N--CH.sub.3,
--C(.dbd.)-(C.sub.1-6 alkyl), --C(.dbd.O)O-(C.sub.1-6 alkyl), or
--OC(.dbd.O)-(C.sub.1-6 alkyl); R.sub.3 is C.sub.1-8 alkyl,
--(CH.sub.2).sub.y-(C.sub.6-18 aryl), or --(CH.sub.2).sub.y-(5-7
membered heterocycle), wherein y is 0 to 4, and wherein R.sub.3 is
unsubstituted or substituted with at least one of halogen, hydroxy,
--C(.dbd.O)-(C.sub.1-6 alkyl), --CN, --C(.dbd.O)O-(C.sub.1-6
alkyl), --OC(.dbd.O)-(C.sub.1-6 alkyl), keto, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6
alkyl)amino; X is hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6
alkoxy, C.sub.1-6 alkylamino, or di-(C.sub.1-6 alkyl)amino; Y is
hydrogen, halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkylamino, or di-(C.sub.1-6 alkyl)amino; and n is 0, 1, or 2.
[0060] In a preferred embodiment, methods are provided for reducing
MIF activity in a patient in need thereof by administering to the
patient an effective amount of a compound having the following
structures (I), (II), and (III): ##STR96## including forms such as
stereoisomers, free forms, pharmaceutically acceptable salts, or
esters thereof, solvates, and combinations of such forms; wherein
n, R.sub.1, R.sub.2, R.sub.3, X, and Y are as defined above.
[0061] As used herein, the above terms have the following meanings.
The term "alkyl," as used herein is a broad term and is used in its
ordinary sense, including, without limitation, to refer to a
straight chain or branched, acyclic or cyclic, unsaturated or
saturated aliphatic hydrocarbon containing 1, 2, 3, 4, 5, 6, 7, or
8 or more carbon atoms (e.g., C.sub.1-8 alkyl), while the term
"lower alkyl" has the same meaning as alkyl but contains 1, 2, 3,
4, 5, or 6 carbon atoms (e.g., C.sub.1-6 alkyl). Representative
saturated straight chain alkyls include methyl, ethyl, n-propyl,
n-butyl, n-pentyl, n-hexyl, and the like; while saturated branched
alkyls include isopropyl, sec-butyl, isobutyl, tert-butyl,
isopentyl, and the like. Unsaturated alkyls contain at least one
double or triple bond between adjacent carbon atoms (referred to as
an "alkenyl" or "alkynyl," respectively).
[0062] The term "cycloalkyl," as used herein is a broad term and is
used in its ordinary sense, including, without limitation, to refer
to alkyls that include mono-, di-, or poly-homocyclic alkyl ring
systems. Cycloalkyls are also referred to as "cyclic alkyls" or
"homocyclic rings." Representative cycloalkyls include cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, --CH.sub.2-cyclopropyl,
--CH.sub.2-cyclobutyl, --CH.sub.2-cyclopentyl,
--CH.sub.2-cyclohexyl, cyclopentenyl, cyclohexenyt decalin, and
adamantane.
[0063] The term "aryl," as used herein is a broad term and is used
in its ordinary sense, including, without limitation, to refer to
an aromatic carbocyclic moiety such as phenyl or naphthyl,
including mono-, di-, and poly-homocyclic aromatic ring systems
(e.g., C.sub.6-18 aryl).
[0064] The term "arylalkyl," as used herein is a broad term and is
used in its ordinary sense, including, without limitation, to refer
to an alkyl having at least one alkyl hydrogen atom replaced with
an aryl moiety, such as benzyl or naphthyl. Representative
arylalkyls include --CH.sub.2-(1-naphthyl),
--CH.sub.2-(2-naphthyl), --CH.sub.2-(phenyl),
--(CH.sub.2).sub.2-(phenyl), --(CH.sub.2).sub.3-(phenyl), and
--CH-(phenyl).sub.2.
[0065] The terms "heterocycle" and "heterocyclic ring," as used
herein, are broad terms and are used in their ordinary sense,
including, without limitation, to refer to a 5, 6, or 7 membered
monocyclic heterocyclic ring, or a 7, 8, 9, 10, 11, 12, 13, or 14
or more membered polycyclic heterocyclic ring. The ring can be
saturated, unsaturated, aromatic (e.g., a heteroaryl), or
nonaromatic, and can contain 1, 2, 3, or 4 or more heteroatoms
independently selected from nitrogen, oxygen, and sulfur. The
nitrogen and sulfur heteroatoms can be optionally oxidized, and the
nitrogen heteroatom can be optionally quaternized, including
bicyclic rings in which any of the above heterocycles are fused to
(or anellated with, spiro-linked to, or bridged to) a phenyl or
naphthyl ring as well as tricyclic (and higher) homocyclic or
heterocyclic ring systems. The heterocycle can be attached to the
remainder of the molecule via any heteroatom or carbon atom of the
ring or rings. Representative heteroaryls include furyl,
benzofuranyl, thiophenyl, benzothiophenyl, pyrrolyl, indolyl,
isoindolyl, azaindolyl, pyridyl, quinolinyl, isoquinolinyl,
oxazolyl, isooxazolyl, benzoxazolyl, pyrazolyl, imidazolyl,
benzimidazolyl, thiazolyl, benzothiazolyl, isothiazolyl,
pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, cinnolinyl,
phthalazinyl, and quinazolinyl. Representative heterocycles also
include morpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl,
hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl,
tetrahydropyranyl, tetrahydropyridinyl, tetrahydroprimidinyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydropyrimidinyl,
tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like. Also
included are heterocycles of the following structures:
##STR97##
[0066] The term "heterocyclealkyl," as used herein is a broad term
and is used in its ordinary sense, including, without limitation,
to refer to an alkyl having at least one alkyl hydrogen atom
replaced with a heterocycle, such as --CH.sub.2-morpholinyl, and
the like.
[0067] The term "substituted," as used herein is a broad term and
is used in its ordinary sense, including, without limitation, to
refer to any of the above groups (e.g., alkyl, aryl, arylalkyl,
heterocycle, or heterocyclealkyl) wherein at least one hydrogen
atom is replaced with a substituent. In the case of a keto
substituent (i.e., --C(.dbd.O)--) two hydrogen atoms are replaced.
Representative substituents within the context of preferred
embodiments include halogen, hydroxy, cyano, nitro, amino,
alkylamino, dialkylamino, alkyl, alkoxy, alkylthio, aryl, and
heterocycle. Particularly preferred substituents include halogen,
C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 alkylamino, and
di-(C.sub.1-6 alkyl)amino.
[0068] The term "halogen," as used herein is a broad term and is
used in its ordinary sense, including, without limitation, to refer
to fluoro (F), chloro (Cl), bromo (Br), and iodo (I).
[0069] The term "alkoxy," as used herein is a broad term and is
used in its ordinary sense, including, without limitation, to refer
to an alkyl moiety attached through an oxygen bridge (i.e.,
--O-alkyl) such as methoxy, ethoxy, and the like.
[0070] The terms "alkylamino" and "dialkylamino" as used herein,
are broad terms and are used in their ordinary sense, including,
without limitation, to refer to one alkyl moiety or two alkyl
moieties, respectively, attached through a nitrogen bridge (e.g.,
--N-alkyl).sub.2 or --N-alkyl). Representative alkylamino and
dialkylamino groups include methylamino, ethylamino, dimethylamino,
diethylamino, and the like.
[0071] The cyclic systems referred to herein include fused ring,
bridged ring, and spiro ring moieties, in addition to isolated
monocyclic moieties.
MIF as a Drug Target
[0072] Macrophage migration inhibitory factor (MIF) is well suited
for analysis as a drug target as its activity has been implicated
in a variety of pathophysiological conditions. For instance, MIF
has been shown to be a significant mediator in both inflammatory
responses and cellular proliferation. In this regard, MIF has been
shown to play roles as a cytokine, a pituitary hormone, as
glucocorticoid-induced immunomodulator, and as a
neuroimmunomodulator and in neuronal function. Takahashi et al.,
Mol. Med. 4:707-714, 1998; Bucala, Ann. N.Y. Acad. Sci. 840:74-82,
1998; Bacher et al., Mol. Med. 4(4):217-230, 1998. Further, it has
been recently demonstrated that anti-MIF antibodies have a variety
of uses, notably decreased tumor growth, along with an observed
reduction in angiogenesis. Ogawa et al., Cytokine 12(4):309-314,
2000; Metz and Bucala (supra). Accordingly, small molecules that
can inhibit MIF have significant value in the treatment of
inflammatory responses, reduction of angiogenesis, viral infection,
bacterial infection, treatment of cancer (specifically
tumorigenesis and apoptosis), treatment of graft versus host
disease and associated tissue rejection. A MIF inhibitor may be
particularly useful in a variety of immune related responses, tumor
growth, glomerulonephritis, inflammation, malarial anemia, septic
shock, tumor associated angiogenesis, vitreoretinopathy, psoriasis,
graft versus host disease (tissue rejection), atopic dermatitis,
rheumatoid arthritis, inflammatory bowel disease, inflammatory lung
disorders, otitis media, Crohn's disease, acute respiratory
distress syndrome, delayed-type hypersensitivity. A MIF inhibitor
may also be useful in the treatment of stress and glucocorticoid
function disorders, e.g., counter regulation of glucocorticoid
action; or overriding of glucocorticoid mediated suppression of
arachidonate release (Cys-60 based catalytic MIF oxidoreductase
activity or JABI/CSNS-MIF-interaction based mechanism). MIF
inhibitors may also be useful in the treatment of systemic lupus
erythematosus (SLE). MIF mRNA expression in peripheral blood
mononuclear cells (PBMC) and serum MIF concentration is
significantly increased in patients with SLE and correlates with
SLE disease activity. See Chen et al., Zhonghua Nei Ke Za Zhi. 2004
August;43(8):572-5.
[0073] While not wishing to be limited to any particular theory of
operation, MIF may likely be produced by activated T-cells and
macrophages during the proinflammatory stage of endotoxin-induced
shock, e.g., as part of the localized response to infection. Once
released by a pro-inflammatory stimulus, e.g., low concentrations
of LPS, or by TNF-.alpha. and IFN-.gamma., macrophage-derived MIF
may be the probable source of MIF produced during the acute phase
of endotoxic shock. Both the pituitary, which releases MIF in
response to LPS, and macrophages are the probable source of MIF in
the post-acute phase of endotoxic shock, when the infection is no
longer confined to a localized site. See, e.g., U.S. Pat. No.
6,080,407, incorporated herein by reference in its entirety and
describing these results with anti-MIF antibodies.
[0074] A variety of inflammatory conditions may be amenable to
treatment with a MIF inhibitor. In this regard, among other
advantages, the inhibition of MIF activity and/or release may be
employed to treat inflammatory response and shock. Beneficial
effects may be achieved by intervention at both early and late
stages of the shock response. In this respect, while not limited to
any theory or mechanism responsible for the protective effect of
MIF inhibition, anti-MIF studies have demonstrated that
introduction of anti-MIF antibodies is associated with an
appreciable (up to 35-40%) reduction in circulating serum
TNF-.alpha.0 levels. This reduction is consistent with the
TNF-a-inducing activity of MIF on macrophages in vitro, and
suggests that MIF may be responsible, in part, for the extremely
high peak in serum TNF-.alpha. level that occurs 1-2 hours after
endotoxin administration despite the fact that MIF cannot be
detected in the circulation at this time. Thus, MIF inhibition
therapy may be beneficial at the early stages of the inflammatory
response.
[0075] MIF also plays a role during the post-acute stage of the
shock response, and therefore, offers an opportunity to intervene
at late stages where other treatments, such as anti-TNF-.alpha.
therapy, are ineffective. Inhibition of MIF can protect against
lethal shock in animals challenged with high concentrations of
endotoxin (i.e., concentrations that induce release of pituitary
MIF into the circulation), and in animals challenged with
TNF-.alpha.. Accordingly, the ability to inhibit MIF and protect
animals challenged with TNF indicates that neutralization of MIF
during the later, post-acute phase of septic shock may be
efficacious.
[0076] As evidenced herein, TNF-.alpha. and IL-1.beta. levels are
correlated at least in some instances to MIF levels. Accordingly,
an anti-MIF small molecule may be useful in a variety of
TNF-.alpha. and/or IL-1.beta., associated disease states including
transplant rejection, immune-mediated and inflammatory elements of
CNS disease (e.g., Alzheimer's, Parkinson's, multiple sclerosis,
and the like), muscular dystrophy, diseases of hemostasis (e.g.,
coagulopathy, veno occlusive diseases, and the like), allergic
neuritis, granuloma, diabetes, graft versus host disease, chronic
renal damage, alopecia (hair loss), acute pancreatitis, joint
disease, congestive heart failure, cardiovascular disease
(restenosis, atherosclerosis), joint disease, and osteoarthritis.
See also Colby-Germinario, et al., J. Neurological Sci., 1977,
33:111-129; Sheremata, et al., J. Neurological Sci., 1978,
36:165-170; Wettinger, et al., Blood. 2005 March 1;105(5):2000-6.
Epub 2004 November 02; and "Chemists say they have identified a
gene that appears to play a key role in the development of type 1
diabetes," Medical Research News, Published: Monday, 21-Mar-2005,
www.news-medical.net.htm.
[0077] Further, additional evidence in the art has indicated that
steroids, while potent inhibitors of cytokine production, actually
increase MIF expression. Yang et al., Mol. Med. 4(6):413-424, 1998;
Mitchell et al., J. Biol. Chem. 274(25):18100-18106, 1999; Calandra
and Bucala, Crit. Rev. Immunol. 17(1):77-88, 1997; Bucala, FASEB J.
10(14):1607-1613, 1996. Accordingly, it may be of particular
utility to utilize MIF inhibitors in combination with steroidal
therapy for the treatment of cytokine mediated pathophysiological
conditions, such as inflammation, shock, and other
cytokine-mediated pathological states, particularly in chronic
inflammatory states such as arthritis, particularly rheumatoid
arthritis. Such combination therapy may be beneficial even
subsequent to the onset of pathogenic or other inflammatory
responses. For example, in the clinical setting, the administration
of steroids subsequent to the onset of septic shock symptoms has
proven of little benefit. See Bone et al., N. Engl. J. Med 317:
653-658, 1987; Spring et al., N. Engl. J. Med 311: 1137-1141, 1984.
Combination steroid/MIF inhibition therapy may be employed to
overcome this obstacle. Further, one of skill in the art may
understand that such therapies may be tailored to inhibit MIF
release and/or activity locally and/or systemically.
Applications and Methods Utilizing MIF Inhibitors
[0078] MIF inhibitors have a variety of applicable uses, as noted
above. Candidate MIF inhibitors may be isolated or procured from a
variety of sources, such as bacteria, fungi, plants, parasites,
libraries of chemicals (small molecules), peptides or peptide
derivatives and the like. Further, one of skill in the art will
recognize that inhibition has occurred when a statistically
significant variation from control levels is observed.
[0079] Given the various roles of MIF in pathology and homeostasis,
inhibition of MIF activity or MIF extracellular localization may
have a therapeutic effect. For example, recent studies have
demonstrated that MIF is a mediator of endotoxemia, where anti-MIF
antibodies fully protected mice from LPS-induced lethality. See
Bernhagen et al., Nature 365:756-759, 1993; Calandra et al., J.
Exp. Med. 179:1895-1902, 1994; Bernhagen et al., Trends Microbiol.
2:198-201, 1994. Further, anti-MIF antibodies have markedly
increased survival in mice challenged with gram-positive bacteria
that induces septic shock. Bernhagen et al., J. Mol. Med.
76:151-161, 1998. Other studies have demonstrated the role of MIF
in tumor cell growth and that anti-sense inhibition of MIF leads to
resistance to apoptotic stimuli. Takahashi et al., Mol. Med.
4:707-714, 1998; Takahashi et al., Microbiol. Immunol. 43(1):61-67,
1999. In addition, the finding that MIF is a counterregulator of
glucocorticoid action indicates that methods of inhibiting MIF
extracellular localization may allow for treatment of a variety of
pathological conditions, including autoimmunity, inflammation,
endotoxemia, and adult respiratory distress syndrome, inflammatory
bowel disease, otitis media, inflammatory joint disease and Crohn's
disease. Bernhagen et al., J. Mol. Med. 76:151-161, 1998; Calandra
et al., Nature 377:68-71, 1995; Donnelly et al., Nat. Med.
3:320-323, 1997. Because MIF is also recognized to be angiogenic,
the inhibition of this cytokine may have anti-angiogenic activity
and particular utility in angiogenic diseases that include, but are
not limited to, cancer, diabetic retinopathy, psoriasis,
inflammation of the skin, angiopathies, fertility, obesity and
genetic diseases of glucocorticoid dysfunction like Cushing's and
Addison's disease. MIF inhibitors may also be useful in treating
conditions such as metabolic syndrome.
[0080] The compounds of the preferred embodiments can be used for
the treatment of a patient that has, or is at risk for having,
diabetes mellitus (e.g., type 1 diabetes, type 2 diabetes,
gestational diabetes), abnormal glucose tolerance, stress
hyperglycemia, metabolic syndrome, and/or insulin resistance. The
compounds of preferred embodiments are especially preferred for
treating a patient having or at risk for type 1 diabetes.
[0081] Type 1 diabetes mellitus is a multifactorial syndrome caused
by the lack of endogenous insulin, thought to be due to an immune
attack mediated by autoreactive T cells and macrophages against
pancreatic p-cells. Extensive research efforts have greatly
expanded understanding of disease pathogenesis, and have revealed a
critical role for several pro-inflammatory mediators. However, no
effective anti-inflammatory therapeutic has been approved for the
clinical management of type 1 diabetes. Several animal models of
the disease have enhanced understanding of the molecular events
that underlie the pathogenesis of diabetes. Multiple low doses of
streptozotocin to susceptible strains of mice induce a diabetic
condition with many of the hallmarks of human type 1 diabetes.
Clinical and histoimmunological similarities include the
development of hyperglycemia associated with infiltration of the
pancreatic islets by T lymphocytes and macrophages (insulitis)
(Like et al., Science 193:415-417, 1976; Kolb, Diabetes Rev.
1:116-126, 1993). Proinflammatory cytokines, including interleukin
(IL)-1.beta., interferon (IFN)-.gamma., tumor necrosis factor
(TNF)-.alpha.and IL-18 play important roles in the development of
streptozotocin-induced diabetes (Sandberg et al., Biochem. Biophys.
Res. Comm. 202:543-548, 1994; Herold et al., J. Immunol.
156:3521-3527, 1996; Holdstad et al., J. Autoimmun. 16:441-447,
2001; Nicoletti et al., Eur. J. Immunol. 33:2278-2286, 2003).
However, administration of either recombinant IL-1.beta.,
IFN-.gamma., or TNF-.beta., or specific inhibitors of their
activity, have complex and often contradictory effects on disease
development and/or course, depending on animal model used, as well
as on timing of administration (Rabinovitch et al., Biochem.
Pharmacol. 55:1139-1149, 1998; Campbell et al., J. Clin. Invest.
87:739-740, 1991; Nicoletti et al., Diabetes 47:32-38, 1998; Yang
et al., J. Exp. Med. 180:995-1004, 1994).
[0082] The key pathogenic role played by the immune system in the
pathogenesis of type 1 diabetes has recently focused on identifying
immunotherapeutical approaches that may allow halting or delaying
P-cell destruction in prediabetic individuals or in those patients
with newly diagnosed disease (Winter et al., Biodrugs 17:39 64,
2003). Macrophage migration inhibitory factor (MIF) is a critical
cytokine in local and systemic inflammation, but its role in
diabetes has not been explored thoroughly. MIF is a pleiotropic
cytokine produced during immune responses by activated T cells,
macrophages and a variety of nonimmune cells (Bucala, FASEB J.
10:1607-1613, 1996; Metz et al., Adv. Immunol. 66: 197-223, 1997).
It acts as a critical mediator of host defense, and is being
explored as a therapeutic target in septic shock as well as chronic
inflammatory and autoimmune diseases (Calandra et al., Nat. Med.
6:164-170, 2000; De Yong et al., Nat. Immunol. 2:1061-1066, 2001;
Denkinger et al., J. Immunol. 170:1274-1282, 2003). Elevated MIF
gene expression has been detected in spontaneously non-obese
diabetic (NOD) mice (Bojunga et al., Cytokine 91:179-186, 2003),
but its importance in the pathogenesis of type 1 diabetes is
unclear. The role of MIF in type 2 diabetes has also been
investigated (Yabunaka N, et al., Diabetes Care 23(2):256-, 2000),
as has the role of MIF in the pathogenesis of proliferative
diabetic retinopathy (Mitamura Y, et al., Br. J. Opthalmol.
84:636-639, 2000)
[0083] A potential role for MIF in the development and pathogenesis
of autoimmune mediated diabetes has been implicated in
spontaneously diabetic NOD mice, because expression of MIF mRNA is
significantly increased during disease development, and exogenous
MIF administration increases disease incidence in these animals
(Bojunga et al., Cytokine 91:179-186, 2003). MIF is constitutively
expressed and secreted together with insulin from pancreatic
p-cells, and acts as an autocrine factor to stimulate insulin
release (Waeber et al., Proc. Natl. Acad. Sci. USA 94:4782-4787,
1997). Because induction of insulin secretion is thought to
contribute to immunoinflammatory diabetogenic pathways by favoring
the expression on the .beta.-cells and the presentation to the
immune cells of antigens that are up-regulated when the functional
activity is augmented (Winter et al., Biodrugs 17:39 64, 2003),
this hormonal property could represent an additional important
factor involving endogenous MIF in the initial events of
.beta.-cell dysfunction and destruction. Targeting endogenous MIF
may therefore be a suitable approach for unraveling the role of
this cytokine in the pathogenesis of type 1 diabetes and for
therapeutic and/or prophylactic treatment of the condition.
[0084] Endogenous MIF has been reported to play a role in the
development of murine autoimmune diabetes (PCT International Publ.
No. WO-2005/094338-A1), where progression of MLD-STZ-induced
diabetes was accompanied by up-regulated MIF protein expression
both in pancreatic islets and peripheral cells, and
immunoneutralization of MIF by anti-MIF IgG, or pharmacological
inhibition of MIF activity with ISO-1, attenuated the clinical and
histological manifestations of the disease.
[0085] The MIF inhibitors activity or export may be employed
therapeutically and also utilized in conjunction with a targeting
moiety that binds a cell surface receptor specific to particular
cells. Compositions of preferred embodiments may be formulated for
administration by any conventional route, including enterally
(e.g., buccal, oral, nasal, rectal), parenterally (e.g.,
intravenous, intracranial, intraperitoneal, subcutaneous, or
intramuscular), or topically (e.g., epicutaneous, intranasal, or
intratracheal). Within other embodiments, the compositions
described herein may be administered as part of a sustained release
implant.
[0086] Within yet other embodiments, compositions of preferred
embodiments may be formulized as a lyophilizate, utilizing
appropriate excipients that provide stability as a lyophilizate,
and subsequent to rehydration.
[0087] Pharmaceutical compositions containing the MIF inhibitors of
preferred embodiments can be manufactured according to conventional
methods, e.g., by mixing, granulating, coating, dissolving or
lyophilizing processes.
[0088] In another embodiment, pharmaceutical compositions
containing one or more MIF inhibitors are provided. For the
purposes of administration, the compounds of preferred embodiments
may be formulated as pharmaceutical compositions. Pharmaceutical
compositions of preferred embodiments comprise one or more MIF
inhibitors of preferred embodiments and a pharmaceutically
acceptable carrier and/or diluent. The inhibitor of MIF is present
in the composition in an amount which is effective to treat a
particular disorder, that is, in an amount sufficient to achieve
decreased MIF levels or activity, symptoms, and/or preferably with
acceptable toxicity to the patient. Preferably, the pharmaceutical
compositions of preferred embodiments can include MIF inhibitor(s)
in an amount from less than about 0.5 mg to more than about 1000 mg
per dosage depending upon the route of administration, preferably
from about 0.6, 0.7, 0.8, or 0.9 mg to about 150, 200, 250, 300,
350, 400, 450, 500, 600, 700, 800, or 900 mg, and more preferably
from about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or 25 mg to about
30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 mg.
In certain embodiments, however, lower or higher dosages than those
mentioned above may be preferred. Appropriate concentrations and
dosages can be readily determined by one skilled in the art.
[0089] Pharmaceutically acceptable carriers and/or diluents are
familiar to those skilled in the art. For compositions formulated
as liquid solutions, acceptable carriers and/or diluents include
saline and sterile water, and may optionally include antioxidants,
buffers, bacteriostats, and other common additives. The
compositions can also be formulated as pills, capsules, granules,
tablets (coated or uncoated), (injectable) solutions, solid
solutions, suspensions, dispersions, solid dispersions (e.g., in
the form of ampoules, vials, creams, gels, pastes, inhaler powder,
foams, tinctures, lipsticks, drops, sprays, or suppositories). The
formulation can contain (in addition to one or more MIF inhibitors
and other optional active ingredients) fillers, disintegrators,
flow conditioners, sugars and sweeteners, fragrances,
preservatives, stabilizers, wetting agents, emulsifiers,
solubilizers, salts for regulating osmotic pressure, buffers,
diluents, dispersing and surface-active agents, binders,
lubricants, and/or other pharmaceutical excipients as are known in
the art. One skilled in this art may further formulate the
inhibitor of MIF in an appropriate manner, and in accordance with
accepted practices, such as those described in Remington's
Pharmaceutical Sciences, Gennaro, Ed., Mack Publishing Co., Easton,
Pa. 1990.
[0090] The compounds of structures (I), (II), and (III) may occur
as isomers, racemates, optical isomers, enantiomers, diastereomers,
tautomers, and cis/trans conformers. All such isomeric forms are
included within preferred embodiments, including mixtures thereof.
The compounds of structures (I), (II), and (III) may have chiral
centers, for example, they may contain asymmetric carbon atoms and
may thus exist in the form of enantiomers or diastereoisomers and
mixtures thereof, e.g. racemates. Any asymmetric carbon atom may be
present in the (R)-, (S)- or (R,S)-configuration, preferably in the
(R)- or (S)-configuration. Isomeric mixtures can be separated, as
desired, according to conventional methods to obtain pure
isomers.
[0091] Furthermore, some of the crystalline forms of the compounds
of structures (I), (II), and (III) may exist as polymorphs, which
are included in preferred embodiments. In addition, some of the
compounds of structures (I), (II), and (III) may also form solvates
with water or other organic solvents. Such solvates are similarly
included within the scope of the preferred embodiments.
[0092] In another embodiment, a method is provided for treating a
variety of disorders or illnesses, including inflammatory diseases,
arthritis, immune-related disorders, and the like. Such methods
include administering of a compound of preferred embodiments to a
warm-blooded animal in an amount sufficient to treat the disorder
or illness. Such methods include systemic administration of an
inhibitor of MIF of preferred embodiments, preferably in the form
of a pharmaceutical composition. As used herein, systemic
administration includes oral and parenteral methods of
administration. For oral administration, suitable pharmaceutical
compositions of an inhibitor of MIF include powders, granules,
pills, tablets, and capsules as well as liquids, syrups,
suspensions, and emulsions. These compositions may also include
flavorants, preservatives, suspending, thickening, and emulsifying
agents, and other pharmaceutically acceptable additives. For
parental administration, the compounds of preferred embodiments can
be prepared in aqueous injection solutions that may contain, in
addition to the inhibitor of MIF activity and/or export, buffers,
antioxidants, bacteriostats, and other additives commonly employed
in such solutions.
[0093] As mentioned above, administration of a compound of
preferred embodiments can be employed to treat a wide variety of
disorders or illnesses. In particular, the compounds of preferred
embodiments may be administered to a warm-blooded animal for the
treatment of inflammation, cancer, immune disorders, and the
like.
[0094] MIF inhibiting compounds may be used in combination
therapies with other pharmaceutical compounds. In preferred
embodiments, the MIF inhibiting compound is present in combination
with conventional drugs used to treat diseases or conditions
wherein MIF is pathogenic or wherein MIF plays a pivotal or other
role in the disease process. In particularly preferred embodiments,
pharmaceutical compositions are provided comprising one or more MIF
inhibiting compounds, including, but not limited to compounds of
structures (I), (II), or (III), in combination with one or more
additional pharmaceutical compounds, including, but not limited to
drugs for the treatment of various cancers, asthma or other
respiratory diseases, sepsis, arthritis, inflammatory bowel disease
(IBD), or other inflammatory diseases, immune disorders, or other
diseases or disorders wherein MIF is pathogenic.
[0095] The MIF inhibitors of preferred embodiments can be used for
pharmaceutical treatment alone or in combination with one or more
other pharmaceutically active agents, e.g., such as agents useful
in treating inflammation, tumor growth, or associated diseases.
Such other pharmaceutically active agents include, e.g., steroids,
glucocorticoids, inhibitors of other inflammatory cytokines (e.g.,
anti-TNF.alpha. antibodies, anti-IL-1 antibodies, anti-IFN-.gamma.
antibodies), and other cytokines such as IL-1RA or IL-10, and other
MIF inhibitors.
[0096] Combination therapies can include fixed combinations, in
which two or more pharmaceutically active agents are in the same
formulation; kits, in which two or more pharmaceutically active
agents in separate formulations are sold in the same package, e.g.,
with instructions for co-administration; and free combinations in
which the pharmaceutically active agents are packaged separately,
but instruction for simultaneous or sequential administration are
provided. Other kit components can include diagnostics, assays,
multiple dosage forms for sequential or simultaneous
administration, instructions and materials for reconstituting a
lyophilized or concentrated form of the pharmaceutical composition,
apparatus for administering the pharmaceutically active agents, and
the like.
[0097] In particularly preferred embodiments, one or more MIF
inhibiting compounds are present in combination with one or more
nonsteroidal anti-inflammatory drugs (NSAIDs) or other
pharmaceutical compounds for treating arthritis or other
inflammatory diseases. Preferred compounds include, but are not
limited to, celecoxib; rofecoxib; NSAIDS, for example, aspirin,
celecoxib, choline magnesium trisalicylate, diclofenac potassium,
diclofenac sodium, diflunisal, etodolac, fenoprofen, flurbiprofen,
ibuprofen, indomethacin, ketoprofen, ketorolac, melenamic acid,
nabumetone, naproxen, naproxen sodium, oxaprozin, piroxicam,
rofecoxib, salsalate, sulindac, and tolmetin; and corticosteroids,
for example, cortisone, hydrocortisone, methylprednisolone,
prednisone, prednisolone, betamethesone, beclomethasone
dipropionate, budesonide, dexamethasone sodium phosphate,
flunisolide, fluticasone propionate, triamcinolone acetonide,
betamethasone, fluocinolone, fluocinonide, betamethasone
dipropionate, betamethasone valerate, desonide, desoximetasone,
fluocinolone, triamcinolone, triamcinolone acetonide, clobetasol
propionate, and dexamethasone.
[0098] In particularly preferred embodiments, one or more MIF
inhibiting compounds are present in combination with one or more
beta stimulants, inhalation corticosteroids, antihistamines,
hormones, or other pharmaceutical compounds for treating asthma,
acute respiratory distress, or other respiratory diseases.
Preferred compounds include, but are not limited to, beta
stimulants, for example, commonly prescribed bronchodilators;
inhalation corticosteroids, for example, beclomethasone,
fluticasone, triamcinolone, mometasone, and forms of prednisone
such as prednisone, prednisolone, and methylprednisolone;
antihistamines, for example, azatadine,
carbinoxamine/pseudoephedrine, cetirizine, cyproheptadine,
dexchlorpheniramine, fexofenadine, loratadine, promethazine,
tripelennamine, brompheniramine, cholopheniramine, clemastine,
diphenhydramine; and hormones, for example, epinephrine.
[0099] In particularly preferred embodiments, one or more MIF
inhibiting compounds are present in combination with pharmaceutical
compounds for treating IBD, such as azathioprine or
corticosteroids, in a pharmaceutical composition.
[0100] In particularly preferred embodiments, one or more MIF
inhibiting compounds are present in combination with pharmaceutical
compounds for treating cancer, such as paclitaxel, in a
pharmaceutical composition.
[0101] In particularly preferred embodiments, one or more MIF
inhibiting compounds are present in combination with
immunosuppresive compounds in a pharmaceutical composition. In
particularly preferred embodiments, one or more MIF inhibiting
compounds are present in combination with one or more drugs for
treating an autoimmune disorder, for example, Lyme disease, Lupus
(e.g., Systemic Lupus Erythematosus (SLE)), or Acquired Immune
Deficiency Syndrome (AIDS). Such drugs may include protease
inhibitors, for example, indinavir, amprenavir, saquinavir,
lopinavir, ritonavir, and nelfinavir; nucleoside reverse
transcriptase inhibitors, for example, zidovudine, abacavir,
lamivudine, idanosine, zalcitabine, and stavudine; nucleotide
reverse transcriptase inhibitors, for example, tenofovir disoproxil
fumarate; non nucleoside reverse transcriptase inhibitors, for
example, delavirdine, efavirenz, and nevirapine; biological
response modifiers, for example, etanercept, infliximab, and other
compounds that inhibit or interfere with tumor necrosing factor;
antivirals, for example, amivudine and zidovudine.
[0102] In particularly preferred embodiments, one or more MIF
inhibiting compounds are present in combination with pharmaceutical
compounds for treating sepsis, such as steroids or anti-infective
agents. Examples of steroids include corticosteroids, for example,
cortisone, hydrocortisone, methylprednisolone, prednisone,
prednisolone, betamethesone, beclomethasone dipropionate,
budesonide, dexamethasone sodium phosphate, flunisolide,
fluticasone propionate, triamcinolone acetonide, betamethasone,
fluocinolone, fluocinonide, betamethasone dipropionate,
betamethasone valerate, desonide, desoximetasone, fluocinolone,
triamcinolone, triamcinolone acetonide, clobetasol propionate, and
dexamethasone. Examples of anti-infective agents include
anthelmintics (mebendazole), antibiotics including aminoclycosides
(gentamicin, neomycin, tobramycin), antifungal antibiotics
(amphotericin b, fluconazole, griseofulvin, itraconazole,
ketoconazole, nystatin, micatin, tolnaftate), cephalosporins
(cefaclor, cefazolin, cefotaxime, ceftazidime, ceftriaxone,
cefuroxime, cephalexin), beta-lactam antibiotics (cefotetan,
meropenem), chloramphenicol, macrolides (azithromycin,
clarithromycin, erythromycin), penicillins (penicillin G sodium
salt, amoxicillin, ampicillin, dicloxacillin, nafcillin,
piperacillin, ticarcillin), tetracyclines (doxycycline,
minocycline, tetracycline), bacitracin; clindamycin; colistimethate
sodium; polymyxin b sulfate; vancomycin; antivirals including
acyclovir, amantadine, didanosine, efavirenz, foscarnet,
ganciclovir, indinavir, lamivudine, nelfinavir, ritonavir,
saquinavir, stavudine, valacyclovir, valganciclovir, zidovudine;
quinolones (ciprofloxacin, levofloxacin); sulfonamides
(sulfadiazine, sulfisoxazole); sulfones (dapsone); furazolidone;
metronidazole; pentamidine; sulfanilamidum crystallinum;
gatifloxacin; and sulfamethoxazole/trimethoprim.
[0103] In the treatment of certain diseases, it may be beneficial
to treat the patient with a MIF inhibitor in combination with an
anesthetic, for example, ethanol, bupivacaine, chloroprocaine,
levobupivacaine, lidocaine, mepivacaine, procaine, ropivacaine,
tetracaine, desflurane, isoflurane, ketamine, propofol,
sevoflurane, codeine, fentanyl, hydromorphone, marcaine,
meperidine, methadone, morphine, oxycodone, remifentanil,
sufentanil, butorphanol, nalbuphine, tramadol, benzocaine,
dibucaine, ethyl chloride, xylocaine, and phenazopyridine.
[0104] The compounds of preferred embodiments can generally be
employed as the free acid or the free base. Alternatively, the
compounds of preferred embodiments can preferably be in the form of
acid or base addition salts. The term "pharmaceutically acceptable
salt" of structures (I), (II), and (III) is intended to encompass
any and all acceptable salt forms. While salt forms of the
preferred embodiments are preferably pharmaceutically acceptable
salts, in certain embodiments pharmaceutically unacceptable salts
can be employed (e.g., for preparation, isolation, and/or
purification purposes).
[0105] The compounds of structure (I), (II), and (III) can be made
according to the organic synthesis techniques known to those
skilled in this field, as well as by the representative methods set
forth in the following examples.
Preparation of Compounds of Structure (I)
[0106] A preferred intermediate in the preparation of compound of
structure (I) is
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester, depicted by formula (4) below. To prepare this
intermediate, methyl-3-amino-thiophene-2-carboxylate was reacted
with ethylmalonyl chloride to yield intermediate
3-(2-ethoxycarbonyl-acetylamino)-thiophene-2-carboxylic acid methyl
ester, depicted by formula (1). This intermediate was converted to
7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester, depicted by formula (2), by reacting with sodium
ethoxide, and was then converted into
5,7-dichloro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester,
depicted by formula (3). Hydrolysis of
5,7-dichloro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester,
depicted by formula (3), yielded
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester, depicted by formula (4) as shown in Scheme 1.
##STR98##
[0107] In one method, intermediate
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester, depicted by formula (4), was reacted with
piperazin-1-yl-thiophene-2-yl-methanone to yield
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno[2,3--
b]-pyridine-6-carboxylic acid ethyl ester, depicted by formula (5).
This intermediate was either reacted with an appropriate halide
(R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield the
target compounds of structure (I) with R.sub.2 as ethyl
carboxylate, R.sub.3 as thiophene and R.sub.1 as defined above, as
shown in Scheme 2. ##STR99##
[0108] To yield compounds of structure (I) wherein R.sub.2 is ethyl
carboxylate, R.sub.3 is furan and R.sub.1 is as defined above,
intermediate
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester, depicted by formula (4), was reacted with
1-(2-furyl)-piperazine to yield
5-oxo-7-[4-(furan-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno-
[2,3-b]-pyridine-6-carboxylic acid ethyl ester, depicted by formula
(6). This intermediate was either reacted with an appropriate
halide (R.sub.1--X) or with boronic acid (R.sub.1--B(OH).sub.2) to
yield the target compounds of structure (I), with R.sub.3 as furan
and R.sub.1 as defined above, as shown in Scheme 3. ##STR100##
[0109] In another method, intermediate
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester, depicted by formula (4), was reacted with
tert-butyl-1-piperazine carboxylate to yield
7-(4-tert-butoxycarbonyl-piperazin-1-yl)-5-oxo-4,5-dihydro-thieno[2,3-b]--
pyridine-6-carboxylic acid ethyl ester, depicted by formula (7).
This intermediate was either reacted with an appropriate halide
(R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield the
intermediate of structure (8), which was deprotected and reacted
with an appropriate acid chloride (R.sub.3--COCI) or acid
(R.sub.3--COOH) to yield target compounds of structure (I) with
R.sub.2 as ethyl carboxylate, and R.sub.1 and R.sub.3 as defined
above, as shown in Scheme 4. ##STR101##
[0110] To prepare compounds of structure (I) with R.sub.2 as
carbonitrile, and R.sub.1 and R.sub.3 as defined above,
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile,
depicted by formula (12), was used as a key intermediate. To
prepare this intermediate, methyl-3-amino-thiophene-2-carboxylate
was reacted with methylcyanoacetate to yield intermediate
3-(2-cyano-acetylamino)-thiophene-2-carboxylic acid methyl ester,
depicted by formula (10). This intermediate was converted to
7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile,
depicted by formula (11), by reacting with sodium ethoxide, and was
then converted into
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile,
depicted by formula (12), as shown in Scheme 5. ##STR102##
[0111] To yield a compound of structure (I) with R.sub.2 as
carbonitrile, R.sub.3 as thiophene and R.sub.1 as defined above,
the intermediate
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile
depicted by formula (12) was reacted with
piperazin-1-yl-thiophene-2-yl-methanone to yield
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-th-
ieno[2,3-b]-pyridine-6-carbonitrile, depicted by formula (13),
which was either reacted with an appropriate halide (R.sub.1--X) or
boronic acid (R.sub.1--B(OH).sub.2) to yield the target compounds
of structure (I) with R.sub.2 as carbonitrile, R.sub.3 as
thiophene, and R.sub.1 as defined above as shown in Scheme 6.
##STR103##
[0112] To yield compounds of structure (I), wherein R.sub.2 is
carbonitrile, R.sub.3 is furan, and R.sub.1 is as defined above,
intermediate
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester, depicted by formula (12), was reacted with
1-(2-furyl)-piperazine to yield
5-oxo-7-[4-(furan-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno-
[2,3-b]-pyridine-6-carbonitrile, depicted by formula (14). This
intermediate was either reacted with an appropriate halide
(R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield target
compounds of structure (I) with R.sub.2 as carbonitrile, R.sub.3 as
furan, and R.sub.1 as defined above, as shown in Scheme 7.
##STR104##
[0113] Compounds of structure (I) with R.sub.2 as carbonitrile, and
R.sub.1 and R.sub.3 as defined above were also prepared from
intermediate
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile,
depicted by formula (12). Intermediate
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile,
depicted by formula (12), was reacted with tert-butyl-1-piperazine
carboxylate to yield
7-(4-tert-butoxycarbonyl-piperazin-1-yl)-5-oxo-4,5-dihydro-thieno[2-
,3-b]-pyridine-6-carbonitrile, depicted by formula (15). This
intermediate was either reacted with an appropriate halide
(R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield an
intermediate of structure (16), which was deprotected and reacted
with an appropriate acid chloride (R.sub.3--COCI), or an acid
(R.sub.3--COOH) to yield target compounds of structure (I) with
R.sub.2 as carbonitrile, and R.sub.1 and R.sub.3 as defined above,
as shown in Scheme 8. ##STR105##
[0114] To yield compounds of structure (I) with R.sub.1 as alkyl
benzoic acid, and R.sub.2 and R.sub.3 as defined above, the
corresponding methyl or ethyl esters were prepared as shown in
Scheme 4 and Scheme 8 above and hydrolyzed to the corresponding
acids by BBr.sub.3, as shown in Scheme 9. ##STR106## Preparation of
Compounds of Structure (II)
[0115] Compounds of structure (II) were prepared by using
4-amino-thiophene-3-carboxylic acid methyl ester, as depicted by
formula (21), as a starting material. To make this compound, methyl
thioglycolate was reacted with methyl acrylate to yield
intermediate 3-methoxycarbonylmethylsulfanykpropionic acid methyl
ester, depicted by formula (18), which was cyclized to
4-oxo-tetrahydro-thiophene-3-carboxylic acid methyl ester, depicted
by formula (19). This intermediate was reacted with hydroxylamine
hydrochloride to yield
4-hydroxyimino-tetrahydro-thiophene-3-carboxylic acid methyl ester,
depicted by formula (20), which yielded
4-amino-thiophene-3-carboxylic acid methyl ester hydrochloride,
depicted by formula (21), as shown in Scheme 10. ##STR107##
[0116] A preferred intermediate in the preparation of a compound of
formula (II) is
4-chloro-1,2-dihydro-2-oxo-thieno[3,4-b]pyridine-3-carboxylic acid
ethyl ester, depicted by formula (25) below. To prepare this
intermediate, 4-amino-thiophene-3-carboxylic acid methyl ester
hydrochloride, depicted by formula (21), was reacted with
ethylmalonyl chloride to yield intermediate
4-(2-ethoxycarbonyl-acetylamino)-thiophene-3-carboxylic acid methyl
ester, depicted by formula (22). This intermediate was converted to
7-hydroxy-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
ethyl ester, depicted by formula (23) by reacting with sodium
ethoxide, and was then converted into
5,7-dichloro-2-thia-4-aza-indene-6-carboxylic acid ethyl ester,
depicted by formula (24). Hydrolysis
5,7-dichloro-2-thia-4-aza-indene-6-carboxylic acid ethyl ester,
depicted by formula (24), yielded
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-3-carboxylic acid
ethyl ester, depicted by formula (25), as shown in Scheme 11.
##STR108##
[0117] In one method,
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-3-carboxylic acid
ethyl ester, depicted by formula (25), was reacted with
piperazin-1-yl-thiophene-2-yl-methanone to yield
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2-thia-4-az-
a-indene-6-carboxylic acid ethyl ester, depicted by formula (26).
This intermediate was either reacted with an appropriate halide
(R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield target
compounds of structure (II) with R.sub.2 as ethyl carboxylate,
R.sub.3 as thiophene, and R.sub.1 as defined above, as shown in
Scheme 12. ##STR109##
[0118] To yield compounds of structure (II) where R.sub.2 is ethyl
carboxylate, R.sub.3 is furan, and R.sub.1 is as defined above,
intermediate
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-3-carboxylic acid
ethyl ester, depicted by formula (25), was reacted with
1-(2-furyl)-piperazine to yield
5-oxo-7-[4-(furan-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2-thia-4-aza-in-
dene-6-carboxylic acid ethyl ester, depicted by formula (27), which
was either reacted with an appropriate halide (R.sub.1--X) or
boronic acid (R.sub.1--B(OH).sub.2) to yield compounds of structure
(I) with R.sub.3 as furan and R.sub.1 as defined above, as shown in
Scheme 13. ##STR110##
[0119] In another method, intermediate
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-3-carboxylic acid
ethyl ester, depicted by formula (25), was reacted with
tert-butyl-1-piperazine carboxylate to yield
7-(4-tert-butoxycarbonyl-piperazin-1-yl)-5-oxo-4,5-dihydro-2-thia-4-aza-i-
ndene-6-carboxylic acid ethyl ester, depicted by formula (28). This
intermediate was either reacted with an appropriate halide
(R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield an
intermediate of structure (29), which was deprotected and reacted
with an appropriate acid chloride (R.sub.3--COCl) or acid
(R.sub.3--COOH) to yield target compounds of structure (II), with
R.sub.2 as ethyl carboxylate, and R.sub.1 and R.sub.3 as defined
above, as shown in Scheme 14. ##STR111##
[0120] To prepare compounds of structure (II) with R.sub.2 as
carbonitrile, and R.sub.1 and R.sub.3 as defined above,
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carbonitrile,
depicted by formula (33), was used as a key intermediate. To
prepare this intermediate,
7-hydroxy-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
ethyl ester, depicted by formula (23), was reacted with
cyclohexylamine to yield intermediate
7-hydroxy-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
cyclohexylamide, depicted by formula (31). This intermediate was
converted to 5,7-dichloro-2-thia-4-aza-indene-6-carbonitrile,
depicted by formula (32), by reacting with phosphorous oxychloride,
and was then converted into
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-6-carbonitrile, depicted by
formula (33), as shown in Scheme 15. ##STR112##
[0121] To yield a compound of structure (II) with R.sub.2 as
carbonitrile, R.sub.3 as thiophene, and R.sub.1 as defined above,
the intermediate
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carbonitrile,
depicted by formula (33), was reacted with
piperazin-1-yl-thiophene-2-yl-methanone to yield
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2--
thia-4-aza-indene-6-carbonitrile, depicted by formula (34), which
was either reacted with an appropriate halide (R.sub.1--X) or
boronic acid (R.sub.1--B(OH).sub.2) to yield target compounds of
structure (II) with R.sub.2 as carbonitrile, R.sub.3 as thiophene,
and R.sub.1 as defined above, as shown in Scheme 16. ##STR113##
[0122] To yield compounds of structure (II) wherein R.sub.2 is
carbonitrile, R.sub.3 is furan, and R.sub.1 is as defined above,
intermediate
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carbonitrile,
depicted by formula (33), was reacted with 1-(2-furyl)-piperazine
to yield
5-oxo-7-[4-(furan-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2-thia-4-aza-in-
dene-6-carbonitrile, depicted by formula (35). This intermediate
was either reacted with an appropriate halide (R.sub.1--X) or
boronic acid (R.sub.1--B(OH).sub.2) to yield the target compounds
of structure (II) with R.sub.2 as carbonitrile, R.sub.3 as furan,
and R.sub.1 as defined above, as shown in Scheme 17. ##STR114##
[0123] Compounds of structure (II) with R.sub.2 as carbonitrile,
and R.sub.1 and R.sub.3 as defined above were also prepared from
intermediate
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carbonitrile,
depicted by formula (33). Intermediate
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carbonitrile,
depicted by formula (33), was reacted with tert-butyl-1-piperazine
carboxylate to yield
7-(4-tert-butoxycarbonyl-piperazin-1-yl)-5-oxo-4,5-dihydro-2-thia-4-
-aza-indene-6-carbonitrile, depicted by formula (36). This
intermediate was either reacted with an appropriate halide
(R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield
intermediate of structure (37), which was deprotected and reacted
with an appropriate acid chloride (R.sub.3--COCI) or acid
(R.sub.3--COOH) to yield target compounds of structure (II) with
R.sub.2 as carbonitrile, and R.sub.1 and R.sub.3 as defined above,
as shown in Scheme 18. ##STR115##
[0124] To yield compounds of structure (II) with R.sub.1 as alkyl
benzoic acid, and R.sub.2 and R.sub.3 as defined above, the
corresponding methyl or ethyl esters were prepared as shown in
Scheme 14 or Scheme 18 above, and hydrolyzed to corresponding acids
by BBr.sub.3, as shown in Scheme 19. ##STR116##
[0125] A preferred intermediate in the preparation of compounds of
structure (III) is
4-chloro-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic acid
ethyl ester, depicted by formula (42) below. To prepare this
intermediate, methyl-2-amino-thiophene-3-carboxylate was reacted
with ethylmalonyl chloride to yield intermediate
2-(2-ethoxycarbonyl-acetylamino)-thiophene-3-carboxylic acid methyl
ester, depicted by formula (39). This intermediate was converted to
4-hydroxy-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic acid
ethyl ester, depicted by formula (40), by reacting it with sodium
ethoxide, and then converting it into
4,6-dichloro-thieno[2,3-b]pyridine-5-carboxylic acid ethyl ester,
depicted by formula (41). Hydrolysis of
4,6-dichloro-thieno[2,3-b]pyridine-5-carboxylic acid ethyl ester,
depicted by formula (41), yielded
4-chloro-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic acid
ethyl ester, depicted by formula (42), as shown in Scheme 20.
##STR117##
[0126] In one method, intermediate
4-chloro-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic acid
ethyl ester, depicted by formula (42), was reacted with
piperazin-1-yl-thiophene-2-yl-methanone to yield
6-oxo-4-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-6,7-dihydro-thieno[3,2--
b]pyridine-5-carboxylic acid ethyl ester, depicted by formula (43).
This intermediate was either reacted with an appropriate halide
(R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield the
target compounds of structure (III) with R.sub.2 as ethyl
carboxylate, R.sub.3 as thiophene, and R.sub.1 as defined above, as
shown in scheme 21. ##STR118##
[0127] To yield compounds of structure (III) wherein R.sub.2 is
ethyl carboxylate, R.sub.3 is furan, and R.sub.1 is as defined
above, intermediate
4-chloro-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic acid
ethyl ester, depicted by formula (42), was reacted with
1-(2-furyl)-piperazine to yield
4-[4-(furan-2-carbonyl)-piperazin-1-yl]-6-oxo-6,7-dihydro-thieno-
[3,2-b]-pyridine-5-carboxylic acid ethyl ester, depicted by formula
(44). This intermediate was either reacted with an appropriate
halide (R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield
the target compounds of structure (III) with R.sub.2 as carboxylic
acid, R.sub.3 as furan and R.sub.1 as defined above, as shown in
scheme 23. ##STR119##
[0128] In another method, intermediate
4-chloro-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic acid
ethyl ester, depicted by formula (42), was reacted with
tert-butyl-1-piperazine carboxylate to yield
4-(4-tert-butoxycarbonyl-piperazin-1-yl)-6-oxo-6,7-dihydro-thieno[2,3-b]--
pyridine-5-carboxylic acid ethyl ester, depicted by formula (45).
This intermediate was either reacted with an appropriate halide
(R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield
intermediate of structure (46), which was deprotected and reacted
with an appropriate acid chloride (R.sub.3--COCl) or acid
(R.sub.3--COOH) to yield target compounds of structure (III) with
R.sub.2 as ethyl carboxylate, and R.sub.1 and R.sub.3 as defined
above, as shown in Scheme 24. ##STR120##
[0129] To yield compounds of structure (III) with R.sub.2 as
carbonitrile, and R.sub.1 and R.sub.3 as defined above,
4-chloro-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carbonitrile,
depicted by formula (50), was used as a key intermediate. To
prepare this intermediate,
4-hydroxy-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic acid
ethyl ester, depicted by formula (40), was reacted with
cyclohexylamine to yield intermediate
4-hydroxy-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic acid
cyclohexylamide, depicted by formula (48). This intermediate was
converted to 4,6-dichloro-thieno[2,3-b]pyridine-5-carbonitrile,
depicted by formula (49), by reacting with phosphorous oxychloride,
which was then converted into
4-chloro-6-oxo-6,7-dihydro-2-thieno[2,3-b]pyridine-5-carbonitrile,
depicted by formula (50), as shown in Scheme 25. ##STR121##
[0130] To yield a compound of structure (III) with R.sub.2 as
carbonitrile, R.sub.3 as thiophene, and R.sub.1 as defined above,
the intermediate
4-chloro-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carbonitrile,
depicted by formula (50), was reacted with
piperazin-1-yl-thiophene-2-yl-methanone to yield
6-oxo-4-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-6,7-dihydro-th-
ieno[2,3-b]pyridine-5-carbonitrile, depicted by formula (51). The
compound of formula (51) was either reacted with an appropriate
halide (R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield
the target compounds of structure (III) with R.sub.2 as
carbonitrile, R.sub.3 as thiophene, and R.sub.1 as defined above,
as shown in Scheme 26. ##STR122##
[0131] To yield compounds of structure (III) wherein R.sub.2 is
carbonitrile, R.sub.3 is furan, and R.sub.1 is as defined above,
intermediate
4-chloro-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carbonitrile,
depicted by formula (50), was reacted with 1-(2-furyl)-piperazine
to yield
4-[4-(furan-2-carbonyl)-piperazin-1-yl]-6-oxo-6,7-dihydro-thieno[2,3-b]-p-
yridine-5-carbonitrile, depicted by formula (52). This intermediate
was either reacted with an appropriate halide (R.sub.1--X) or
boronic acid (R.sub.1--B(OH).sub.2) to yield target compounds of
structure (III) with R.sub.2 as carbonitrile, R.sub.3 as furan, and
R.sub.1 as defined above, as shown in Scheme 27. ##STR123##
[0132] Compounds of structure (III) with R.sub.2 as carbonitrile,
and R.sub.1 and R.sub.3 as defined above were also prepared from
intermediate
4-chloro-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carbonitrile,
depicted by formula (50). Intermediate
4-chloro-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carbonitrile,
depicted by formula (50), was reacted with tert-butyl-1-piperazine
carboxylate to yield
4-(5-cyano-6-oxo-6,7-dihydro-thieno[2,3-b]-pyridine-4-yl)-piperazin-
e-1-carboxylic acid tert-butyl ester, depicted by formula (53).
This intermediate was either reacted with an appropriate halide
(R.sub.1--X) or boronic acid (R.sub.1--B(OH).sub.2) to yield an
intermediate of structure (54), which was deprotected and reacted
with an appropriate acid chloride (R.sub.3--COCI) or acid
(R.sub.3--COOH) to yield target compounds of structure (III) with
R.sub.2 as carbonitrile, and R.sub.1 and R.sub.3 as defined above,
as shown in Scheme 28. ##STR124##
[0133] To yield compounds of structure (III) with R.sub.1 as alkyl
benzoic acid, and R.sub.2 and R.sub.3 as defined above,
corresponding methyl or ethyl esters were prepared as shown in
Scheme 24 or Scheme 28 above and hydrolyzed to corresponding acids
by BBr.sub.3, as shown in Scheme 29. ##STR125## Alternative Method
for the Preparation of Compounds of Structure (I)
[0134] In an alternative method for the preparation of compounds of
structure (I) with R.sub.1, R.sub.2, R.sub.3, X, and Y as defined
above, appropriately substituted
1H-thieno[3,2-d][1,3]oxazine-2,4-dione, depicted by formula (54) in
Scheme 30, was used as an intermediate. To prepare this
intermediate, substituted 3-amino-thiophene-2-carboxylic acid ester
was hydrolyzed to the corresponding 3-amino-thiophene-2-carboxylic
acid, which was reacted with trichloromethyl chloroformate as shown
in Scheme 30. Appropriate N-substitution was introduced by reacting
substituted 1H-thieno[3,2-d][1,3]oxazine-2,4-dione, depicted by
formula (54), with the corresponding halide (R.sub.1--X). The
N-substituted intermediate, depicted by formula (55), was then
reacted with dialkyl malonate to yield the intermediate of general
formula (56) which was chlorinated either by phosphorous
oxychloride or by oxalyl chloride, as depicted in Scheme 30.
##STR126##
[0135] The chloro intermediate, depicted by formula (57), was
reacted with piperazine to yield a piperazine intermediate,
depicted by formula (58), as shown in Scheme 31. Acylation of the
piperazine intermediate, depicted by formula (58), by an
appropriate acyl halide (R.sub.3--CO--Cl) or by coupling with an
appropriate acid (R.sub.3--COOH) yielded a compound of general
formula (I) as shown in Scheme 31. Alternatively, the chloro
intermediate was reacted with an acyl piperazine to yield a
compound of general formula (I) with R.sub.1, R.sub.2, R.sub.3, X,
and Y as defined above, as shown in Scheme 31. ##STR127##
Alternative Method for the Preparation of Compounds of Structure
(III)
[0136] In an alternative method for the preparation of compounds of
structure (III) with R.sub.1, R.sub.2, R.sub.3, X, and Y as defined
above, an appropriately substituted
1H-thieno[2,3-d][1,3]oxazine-2,4-dione, depicted by formula (59) in
Scheme 32, was used as an intermediate. To prepare this
intermediate, a substituted 2-amino-thiophene-3-carboxylic acid
ester was hydrolyzed to the corresponding
2-amino-thiophene-3-carboxylic acid, which was then reacted with
trichloromethyl chloroformate as shown in Scheme 32. Appropriate
N-substitution was introduced by reacting a substituted
1H-thieno[2,3-d][1,3]oxazine-2,4-dione, depicted by formula (59),
with a corresponding halide (R.sub.1--X). The N-substituted
intermediate, depicted by formula (60), was then reacted with
dialkyl malonate to yield an intermediate of general formula (61),
which was chlorinated either by phosphorous oxychloride or by
oxalyl chloride. ##STR128##
[0137] The chloro intermediate, depicted by formula (62), was
reacted with piperazine to yield a piperazine intermediate,
depicted by formula (63), as shown in Scheme 33. Acylation of the
piperazine intermediate (63) by an appropriate acyl halide
(R.sub.3--CO--Cl) or by coupling with an appropriate acid
(R.sub.3--COOH) yielded a compound of general formula (III) as
shown in Scheme 33. Alternatively, the chloro intermediate was
reacted with acyl piperazine to yield a compound of general formula
(III), with R.sub.1, R.sub.2, R.sub.3, X, and Y as defined above,
as shown in Scheme 33. ##STR129##
Alternative Method for the Preparation of Intermediate
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester (4)
[0138] The preferred intermediate in the preparation of compounds
of structure (I),
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester depicted by formula (4) below, was also prepared by an
alternative route as shown in Scheme 34. In this method,
methyl-3-amino-thiophene-2-carboxylate was reacted with 4-methoxy
benzylchloride to yield methyl
3-(4-methoxybenzylamino)thiophene-2-carboxylate, depicted by
formula (65). This intermediate was reacted with ethylmalonyl
chloride followed by a cyclization reaction to yield
7-hydroxy-4-(4-methoxybenzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-c-
arboxylic acid ethyl ester, depicted by formula (67). Chlorination
of
7-hydroxy-4-(4-methoxybenzyl)5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-ca-
rboxylic acid ethyl ester, depicted by formula (67) by reacting
with oxalyl chloride followed by deprotection yielded the
intermediate
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester, depicted by formula (4), as shown in Scheme 34.
##STR130## Experiments
[0139] Macrophage migration inhibitory factor inhibitors of
preferred embodiments were prepared by the methods described in
following examples.
Synthesis of
3-(2-ethoxycarbonyl-acetylamino)-thiophene-2-carboxylic acid methyl
ester (1)
[0140] Ethylmalonyl chloride (1.93 mL, 15.22 mmol) was added to a
solution of methyl-3-amino-thiophene-2-carboxylate (2 g, 12.70
mmol) in dry toluene (20 mL) and pyridine (1.23 mL, 15.22 mmol) at
-10.degree. C. The solution was stirred at -10.degree. C. for 1 h
and poured into ice water. The product was extracted by ethyl
acetate. The combined organic phase was sequentially washed by
diluted HCl solution, saturated NaHCO.sub.3 solution, water, and
brine. The organic phase was dried over MgSO.sub.4 and concentrated
to yield an oily residue. The residue was dissolved in hot ethanol
and kept overnight at -4.degree. C. The crystals formed were
filtered off and filtrate was concentrated to yield 2.2 g (63%) of
3-(2-ethoxycarbonyl-acetylamino)-thiophene-2-carboxylic acid methyl
ester as yellow viscous oil. .sup.1H NMR (400 MHz, DMSO-d6) .delta.
1.21 (t, J=7.2 Hz, 3H), 3.68 (s, 2H), 3.84 (s, 3H), 4.15 (q, J=7.2
Hz, 2H), 7.91 (d, J=5.2 Hz, 1H), 7.93 (d, J=5.2 Hz, 1H), 10.52 (s,
1H) ppm; MS m/z=272 amu (M.sup.++1). ##STR131##
Synthesis of
7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester (2)
[0141] Sodium ethoxide (0.67 g, 9.29 mmol) was added to a solution
of 3-(2-ethoxycarbonyl-acetylamino)-thiophene-2-carboxylic acid
methyl ester (1) (2.18 g, 7.74 mmol) in anhydrous ethanol and
refluxed overnight. The solution was cooled and excess solvent was
distilled off. The residue was dissolved in water and acidified by
cold diluted HCl solution. The solids formed were filtered, washed
by water, and dried under vacuum at room temperature to yield 1.0 g
(55%) of
7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester as white solid. MP 218.degree. C.; .sup.1H NMR (400
MHz, DMSO-d6) .delta. 1.28 (t, J=7.2 Hz, 3H), 4.29 (q, J=7.2 Hz,
2H), 6.96 (d, J=5.6 Hz, 1H), 8.07 (d, J=5.6 Hz, 1H) ppm; MS m/z=240
amu (M.sup.++1). Synthesis of
5,7-dichloro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester
(3)
[0142] A solution of
7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester (2) (1 g, 4.18 mmol) in neat phosphorous oxychloride
was heated at 90.degree. C. for 4 h. The solution was cooled and
excess phosphorus oxychloride was distilled under vacuum. The
residue was suspended in water, basified by solid NaHCO.sub.3, and
extracted by ethyl acetate. The organic phase was washed by
saturated NaHCO.sub.3 solution, water, and brine. The organic phase
was dried over MgSO.sub.4 and concentrated to yield
5,7-dichloro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester
(3) as white solids. Yield 0.56 g (52%); MP 73.degree. C.; .sup.1H
NMR (400 MHz, DMSO-d6) .delta.1.35 (t, J=7.2 Hz, 3H), 4.48 (q,
J=7.2 Hz, 2H), 7.73 (d, J=5.6 Hz, 1H), 8.50 (d, J=5.6 Hz, 1H) ppm;
MS m/z=259 amu (M.sup.+).
Synthesis of
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester (4)
[0143] Ammonium acetate (161 mg, 2.1 mmol) was added to a stirred
solution of 5,7-dichloro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester (3) (530 mg, 1.9 mmol) in glacial acetic acid at room
temperature. The solution was heated at 140.degree. C. for 48 h.
The hot solution was poured into ice water. The solids formed were
filtered, washed by water, and dried. The crude product was
recrystallized by CH.sub.2Cl.sub.2 to yield 273 mg (60%) of
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester as white solids. MP 206.degree. C.; .sup.1H NMR (400
MHz, DMSO-d6) .delta. 1.28 (t, J=7.2 Hz, 3H), 4.29 (q, J=7.2 Hz,
2H), 7.10 (d, J=5.2 Hz, 1H), 8.13 (d, J=5.2 Hz, 1H) ppm; MS m/z=240
amu (M.sup.++1). ##STR132##
Synthesis of
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno[3,2--
b]pyridine-6-carboxylic acid ethyl ester (5)
[0144] 1,4-Diazabicyclo[2.2.2]octane (224 mg, 2 mmol) was added to
a solution of
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester (4) (240 mg, 1 mmol) and
piperazin-1-yl-thiophene-2-yl-methanone (215 mg, 1.1 mmol) in dry
DMA. The solution was heated at 120.degree. C. for 2 h. The
solution was cooled and poured into ice water. The solids formed
were filtered, washed by water, and dried to yield
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno[3,2--
b]pyridine-6-carboxylic acid ethyl ester (400 mg, 96%) as white
solids. MP 252.degree. C.; 1H NMR (400 MHz, DMSO-d6) .delta. 1.27
(t, J=7.2 Hz, 3H), 3.31 (m, 4H), 3.77 (m, 4H), 4.24 (q, J=7.2 Hz,
2H), 6.97 (d, J=5.6 Hz, 1H), 7.15 (dd, J=3.6, 4.8 Hz, 1H), 7.47
(dd, J=1.2, 3.6 Hz, 1H), 7.80 (dd, J=1.2, 4.8 Hz, 1H), 7.96 (d,
J=5.6 Hz, 1H), 12.10 (s, 1H) ppm; MS m/z=418 amu (M.sup.++1).
##STR133##
Synthesis of
4-(4-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl1-4,5-
-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester
(6)
[0145] A solution of
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno[3,2--
b]pyridine-6-carboxylic acid ethyl ester (5) (232 mg, 0.55 mmol) in
dry DMF was added to a suspension of NaH (60% in mineral oil, 24
mg, 0.61 mmol) at room temperature. The solution was stirred at
room temperature for 15 min. 4-Fluorobenzylbromide (76 .mu.L, 0.61
mmol) was added to the solution through a syringe and further
stirred at room temperature for 1 h. The solution was poured into
ice water and the solids formed were filtered, washed by cold
water, and dried. The crude product was purified by flash
chromatography eluting with 0-2% methanol in CH.sub.2Cl.sub.2
gradient to yield 207 mg (90%) of
4-(4-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]4,5--
dihydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester as
white solids. .sup.1H NMR (400 MHz, DMSO-d6) .delta. 1.28 (t, J=7.2
Hz, 3H), 3.34 (m, 4H), 3.80 (m, 4H), 4.27 (q, J=7.2 Hz, 2H), 5.34
(s, 2H), 7.14-7.18 (m, 3H), 7.29-7.35 (m, 3H), 7.48 (dd, J=1.2,
3.6, Hz, 1H), 7.80 (dd, J=1.2, 5.2 Hz, 1H), 8.04 (d, J=5.6 Hz, 1H)
ppm; MS m/z=526 amu (M.sup.++1). Anal.
(C.sub.26H.sub.24FN.sub.3O.sub.4S.sub.2) C, H, N.
Synthesis of
5-oxo-4-(2-oxo-2-phenyl-ethyl)-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl-
]-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester
(7)
[0146] A solution of
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno[3,2--
b]pyridine-6-carboxylic acid ethyl ester (5) (750 mg, 1.79 mmol),
Cs2CO3 (1.76 g, 5.40 mmol), 2-chloroacetophenone (0.333 g, 2.15
mmol) in dry NMP was heated overnight at 90.degree. C. The solution
was poured into ice water and the solids formed were filtered,
washed by cold water, and dried. The crude product was purified by
recrystallization with acetone. MP 240.degree. C. 1H-NMR (DMSO-d6)
.delta. 1.26 (t, J=7.2 Hz, 3H), 3.39 (m, 4H), 3.83 (m, 4H), 4.23
(q, J=7.2 Hz, 2H), 4.23 (q, J=7.2 Hz, 2H), 5.75 (s, 2H), 7.16 (m,
1H), 7.34 (d, J=5.6 Hz, 1H), 7.50 (dd, J=1.2, 3.6 Hz, 1H), 7.61 (t,
J=7.6 Hz, 2H), 7.74 (m, 1H), 7.80 (dd, J=1.2, 5.2 Hz, 1H), 8.01 (d,
J=7.0 Hz, 1H), 8.10 (m, 2H); EIMS m/z 536(M+1).
Synthesis of
5-oxo-4-pyridin-3-yl-methyl-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4-
,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester
(8)
[0147] A solution of
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno[3,2--
b]pyridine-6-carboxylic acid ethyl ester (5) (856 mg, 2.05 mmol),
Cs2CO3 (2.75 g, 8.4 mmol), 3-chloromethyl pyridine hydrochloride
(0.370 g, 2.25 mmol) and KI (1 g) in dry NMP was heated overnight
at 90.degree. C. The solution was poured into ice water and the
solids formed were filtered, washed by cold water, and dried. The
crude product was purified by reverse phase flash chromatography in
combiflash eluting with water/acetonitrile gradient. 1H-NMR
(DMSO-d6) .delta. 3.13 (t, J=7.2 Hz, 3H), 3.80 (m, 4H), 4.27 (q,
J=7.2 Hz, 2H), 5.40 (s, 2H), 7.15 (m, 1H), 7.35 (m, 1H), 7.44 (d,
J=6.0 Hz, 1H), 7.48 (dd, J=1.2, 3.6 Hz, 1H), 7.60 (m, 1H), 7.79
(dd, J=0.8, 4.8 Hz, 1H), 8.06 (d, J=5.6 Hz, 1H), 8.47 (dd, J=1.6,
4.8 Hz, 1H), 8.56 (d, J=1.6 Hz, 1H); EIMS m/z 509 (M+1).
Synthesis of
4-(3-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-
-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester
(9)
[0148] A solution of
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno[3,2--
b]pyridine-6-carboxylic acid ethyl ester (5) (2 g, 4.79 mmol) in
dry DMF was added to a suspension of NaH (60% in mineral oil, 230
mg, 5.74 mmol) at room temperature. The solution was stirred at
room temperature for 30 min. 3-Fluorobenzylbromide (0.705 mL, 5.74
mmol) was added to the solution through a syringe and the solution
was further stirred at room temperature for 3 h. The solvent was
evaporated under vacuum and the residue was suspended in water,
sonicated briefly, and filtered. The solids were washed by cold
water and air dried. The crude product was purified by flash
chromatography eluting with 0-2% methanol in CH.sub.2Cl.sub.2
gradient to yield 1.2 g (48%) of
4-(3-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-
-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester as
white solids. MP 95-113.degree. C. .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.29 (t, J=7.2Hz, 3H), 3.37 (m, 4H), 3.82 (m, 4H), 4.27 (q,
J=7.2 Hz, 2H), 5.38 (s, 2H), 7.05-7.16 (m, 4H), 7.33 (d, J=5.6 Hz,
1H), 7.36 (m, 1H), 7.48 (dd, J=1.2, 3.6 Hz, 1H), 7.79 (dd, J=1.2,
5.2 Hz, 1H), 8.03 (d, J=5.6 Hz, 1H); EIMS m/z 526 (M+1). Anal.
(C.sub.26H.sub.24FN.sub.3O.sub.4S.sub.2) C, H, N.
Synthesis of
7-[4-(furan-2-carbonyl)-piperazin-1-yl]-5-oxo-4,5-dihydro-thieno[3,2-b]py-
ridine-6-carboxylic acid ethyl ester (10)
[0149] 1,4-Diazabicyclo[2.2.2]octane (3.08 g, 27.45 mmol) was added
to a solution of
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester (4) (3.54 mg, 13.77 mmol) and 1-(2-furayl)-piperazine
(3.22 g, 17.87 mmol) in dry DMA. The solution was heated at
120.degree. C. for 2 h. The solution was cooled and poured into ice
water. The solids formed were filtered, washed by water, and dried
to yield
7-[4-(furan-2-carbonyl)-piperazin-1-yl]-5-oxo-4,5-dihydro-thieno[3,2-b]py-
ridine-6-carboxylic acid ethyl ester as brown solid. Yield 5.86 g
(99%). .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.26 (t, J=7.2 Hz, 3H),
3.34 (m, 4H), 3.80 (br, 4H), 4.23 (q, J=7.2 Hz, 2H), 6.65 (m, 1H),
6.97 (d, J=5.6 Hz, 1H), 7.06 (dd, J=0.8, 3.2 Hz, 1H), 7.87 (dd,
J=0.8, 2.0 Hz, 1H), 7.96 (d, J=5.2 Hz, 1H), 12.09 (br, 1H); EIMS
m/z 402 (M+1).
Synthesis of
4-(4-fluoro-benzyl)-7-[4-(furan-2-carbonyl)-piperazin-1-yl]-5-oxo-4,5-dih-
ydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester (11)
[0150] A solution of
7-[4-(furan-2-carbonyl)-piperazin-1-yl]-5-oxo-4,5-dihydro-thieno[3,2-b]py-
ridine-6-carboxylic acid ethyl ester (10) (750 mg, 1.87 mmol),
Cs.sub.2CO.sub.3 (1.83 g, 5.63 mmol), 4-fluoro benzyl bromide
(0.276 mL, 2.24 mmol) in dry NMP was heated overnight at 90.degree.
C. The solution was poured into ice water and the solids formed
were filtered, washed by cold water, and dried. The crude product
was purified by reverse phase flash chromatography in combiflash
eluting with water/acetonitrile gradient. Yield 240 mg (25%).
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.27 (t, J=6.8 Hz, 3H), 3.36 (m,
4H), 3.82 (m, 4H), 4.26 (q, J=6.8 Hz, 2H), 5.35 (s, 2H), 6.65 (m,
1H), 7.06 (dd, J=0.8, 3.6 Hz, 1H), 7.16 (m, 2H), 7.32 (m, 3H), 7.87
(m, 1H), 8.03 (d, J=5.6 Hz, 1H); EIMS m/z 510 (M+1).
Synthesis of
4-(3-fluoro-benzyl)-7-[4-(furan-2-carbonyl)-piperazin-1-yl]-5-oxo-4,5-dih-
ydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester (12)
[0151] This compound was prepared from
7-[4-(furan-2-carbonyl)-piperazin-1-yl]-5-oxo-4,5-dihydro-thieno[3,2-b]py-
ridine-6-carboxylic acid ethyl ester (10) and 3-fluoro benzyl
bromide by applying a similar method as described for
4-(4-fluoro-benzyl)-7-[4-(furan-2-carbonyl)-piperazin-1-yl]-5-oxo-4,5-dih-
ydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester (11).
Yield 210 mg, (22%). .sup.1H-NMR (DMSO-d.sub.6): .delta. 1.28 (t,
J=7.2 Hz, 3H), 3.82 (m, 4H), 4.26 (q, J=7.2 Hz, 2H), 5.38 (s, 2H),
6.65 (m, 1H), 7.07 (m, 4H), 7.34 (d, J=5.6 Hz, 1H), 7.37 (m, 1H),
7.88 (m, 1H), 8.03 (d, J=5.6 Hz, 1H); EIMS m/z 510 (M+1).
Synthesis
7-[4-(furan-2-carbonyl)-piperazin-1-yl]-5-oxo-4-(2-oxo-2-phenyl--
ethyl)-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl
ester (13)
[0152] This compound was prepared from
7-[4-(furan-2-carbonyl)-piperazin-1-yl]-5-oxo-4,5-dihydro-thieno[3,2-b]py-
ridine-6-carboxylic acid ethyl ester (10) and 2-chloroacetophenone
by applying similar method as described for
4-(4-fluoro-benzyl)-7-[4-(furan-2-carbonyl)-piperazin-1-yl]-5-oxo-4,5-dih-
ydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester (11). MP
246.degree. C. .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.25 (t, J=7.2
Hz, 3H), 3.85 (m, 4H), 4.23 (q, J=6.8 Hz, 2H), 5.75 (s, 2H), 6.66
(m, 1H), 7.07 (d, J=3.2 Hz, 1H), 7.34 (d, J=5.6 Hz, 1H), 7.61 (t,
J=7.6 Hz, 2H), 7.73 (t, J=7.6 Hz, 1H), 7.88 (d, J=0.8 Hz, 1H), 8.01
(d, J=5.6 Hz, 1H), 8.10 (d, J=7.6 Hz, 2H); EIMS m/z 520 (M+1).
##STR134##
Synthesis of 1H-thieno3,2-d][1,3]oxazine-2,4-dione (14)
[0153] Methyl 3-aminothiophene-2-carboxylate (31.0 g, 0.20 mol) was
added to a solution of potassium hydroxide (22.68 g, 0.40 mol) in
1L water. The solution was heated at 90.degree. C. for 2 hours. The
solution was then cooled to 0.degree. C. and trichloromethyl
chloroformate (35.7 mL, 0.30 mol) was added slowly, maintaining the
temperature between 0.degree. C. and 10.degree. C. The solution was
stirred at 0.degree. C. for 4 hours, and then allowed to warm up
gradually to room temperature and stirred for another 3 hours. The
precipitated solid product was collected by vacuum filtration to
give 28.5 g (85% yield) 1H-thieno[3,2-d][1,3]oxazine-2,4-dione.
.sup.1H-NMR (DMSO-d.sub.6) .delta. 6.94 (d, J=5.0 Hz, 1H), 8.24 (d,
J=5.0 Hz, 1H), 12.26 (b, 1H) ppm; EIMS m/z 170 (M+1).
Synthesis of
4-(3-fluoro-benzyl)-7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-c-
arboxylic acid ethyl ester (15)
[0154] 1H-Thieno[3,2-d][1,3]oxazine-2,4-dione (14) (40.0 g, 0.24
mol) was added to a suspension of sodium hydride (60% dispersion in
mineral oil, 21.75 g, 0.54 mol) in 300 mL anhydrous DMF stirred
under argon at -10.degree. C. After stirring at -10.degree. C. for
15 minutes, 3-fluorobenzylbromide (29.73 mL, 0.24 mol) was added to
the solution. The solution was allowed to come to room temperature
and further stirred for 3 hours. The solution was again cooled to
-10.degree. C., and diethylmalonate (36.62 mL, 0.24 mol) was added
slowly. The solution was then heated at 110.degree. C. for 45
minutes. A large amount of gas evolved very quickly once the
solution was heated, so this heating step was done in a flask at
least 5 times as large as the reaction volume, and a reflux
condenser was used which was open to the air, and not sealed with a
septum. The reaction mixture was cooled to room temperature, and
poured into a solution of potassium carbonate (32.68 g, 0.24 mol)
in 2.5 L of water. This aqueous solution was stirred for 5 minutes,
and then it was extracted 2 times with 600 mL ethyl acetate and 2
times with 600 mL isopropyl ether. These organic phases were
discarded. The aqueous solution was then acidified slowly to pH 2
with 4M HCl. The precipitated solid product was collected by vacuum
filtration to yield 60.1 g (73% yield) of
4-(3-fluoro-benzyl)-7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]py-
ridine-6-carboxylic acid ethyl ester. .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.30 (t, J=7.2 Hz, 3H), 4.32 (q, J=7.2 Hz, 2H), 5.36 (s,
2H), 7.06 (m, 3H), 7.33 (m, 2H), 8.15 (m, 1H), 13.37 (s, 1H) ppm;
EIMS m/z 348 (M+1).
Synthesis of
4-(3-fluoro-benzyl)-7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-c-
arboxylic acid isopropyl ester (16)
[0155] This compound was prepared by using the same procedure as
described for
4-(3-fluoro-benzyl)-7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-c-
arboxylic acid ethyl ester (15) by using diisopropyl malonate.
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.30 (d, J=6.4 Hz, 6H), 5.17 (m,
1H), 5.36 (s, 2H), 7.08 (m, 3H), 7.31 (m, 2H), 8.15 (dd, J=2.4, 5.2
Hz, 1H), 13.41 (b, 1H) ppm; EIMS m/z 362 (M+1).
Synthesis of
7-chloro-4-(3-fluoro-benzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-ca-
rboxylic acid ethyl ester (17)
[0156]
4-(3-Fluoro-benzyl)-7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyrid-
ine-6-carboxylic acid ethyl ester (15) (60 g, 0.17 mol) was
dissolved in 600 mL anhydrous DMF under a blanket of argon and
cooled to -30.degree. C. Oxalyl chloride (40.7 mL, 0.47 mol) was
then added very slowly (producing a large volume of gas), with the
reaction vessel open to the atmosphere. The solution was then
heated to 75.degree. C. for 2 hours. The solution was cooled to
room temperature and poured into a solution of 150 g. NaCl in 6 L
ice water without stirring. The mixture was allowed to sit without
stirring for 15 minutes, and was then stirred vigorously for 1
minute with a spatula. The precipitated solid product was collected
by vacuum filtration to yield 55 g (87% yield) of
7-chloro-4-(3-fluoro-benzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-ca-
rboxylic acid ethyl ester. .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.30
(t, J=7.2 Hz, 3H), 4.34 (q, J=7.2 Hz, 2H), 5.45 (s, 2H), 7.06-7.17
(m, 3H), 7.37 (m, 1H), 7.49 (d, J=5.6 Hz, 1H), 8.20 (d, J=5.6 Hz,
1H) ppm; EIMS m/z 366 (M+1). ##STR135##
Synthesis of
7-chloro-4-(3-fluoro-benzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-ca-
rboxylic acid isopropyl ester (18)
[0157] This compound was prepared from
4-(3-fluoro-benzyl)-7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-c-
arboxylic acid isopropyl ester (16) by applying the same procedure
as described for
7-chloro-4-(3-fluoro-benzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-ca-
rboxylic acid ethyl ester (17). .sup.1H-NMR (DMSO-d.sub.6): .delta.
1.31 (d, J=6.4 Hz, 6H), 5.16 (m, 1H), 5.45 (s, 2H), 7.05-7.16 (m,
3H), 7.38 (m, 1H), 7.49 (d, J=5.2 Hz, 1H), 8.19 (d, J=5.6 Hz, 1H)
ppm; EIMS m/z 380 (M+1).
Synthesis of
4-(3-fluoro-benzyl)-5-oxo-7-piperazin-1-yl-4,5-dihydro-thieno[3,2-b]pyrid-
ine-6-carboxylic acid ethyl ester (19)
[0158]
7-Chloro-4-(3-fluoro-benzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridi-
ne-6-carboxylic acid ethyl ester (17) (10 g, 27.39 mmol) was added
slowly to a solution of piperazine (9.42 g, 109 mmol) in
dichloromethane. The solution was stirred at room temperature for 3
h. The solvent was evaporated under vacuum. The residue was
suspended in water, stirred vigorously at room temperature, and
filtered. The solids were again dissolved in dichloromethane and
washed by water. The organic phase was dried over MgSO.sub.4 and
concentrated to yield 9.63 g (84%) of
4-(3-fluoro-benzyl)-5-oxo-7-piperazin-1-yl-4,5-dihydro-thieno[3,2-b]pyrid-
ine-6-carboxylic acid ethyl ester. .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.29 (m, 3H), 2.81 (m, 4H), 3.21 (m, 4H), 4.25 (m, 2H),
5.35 (s, 2H), 7.12 (m, 3H), 7.30 (m, 2H), 7.97 (m, 1H); EIMS m/z
416 (M+1).
Synthesis of
4-(3-fluoro-benzyl)-5-oxo-7-piperazin-1-yl-4,5-dihydro-thieno[3,2-b]pyrid-
ine-6-carboxylic acid isopropyl ester (20)
[0159] This compound was prepared from
7-chloro-4-(3-fluoro-benzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-ca-
rboxylic acid isopropyl ester (18) by applying the same procedure
as described for
4-(3-fluoro-benzyl)-5-oxo-7-piperazin-1-yl-4,5-dihydro-thieno[3,2-b]pyrid-
ine-6-carboxylic acid ethyl ester (19). .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.28 (d, J=6.0 Hz, 6H), 2.81 (m, 4H), 3.22 (m, 4H), 5.07
(m, 1H), 5.36 (s, 2H), 7.10 (m, 3H), 7.29 (m, 1H), 7.35 (m, 1H),
7.98 (m, 1H); EIMS m/z 430 (M+1).
Synthesis of
4-(3-fluoro-benzyl)-7-[4-(5-fluoro-thiophene-2-carbonyl)-piperazin-1-yl]--
5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid isopropyl
ester (21)
[0160] A solution of
4-(3-fluoro-benzyl)-5-oxo-7-piperazin-1-yl-4,5-dihydro-thieno[3,2-b]pyrid-
ine-6-carboxylic acid isopropyl ester (20) (2 g, 4.65 mmol), HOBt
(0.692 g, 5.12 mmol), EDC-HCl (0.982 g, 5.12), triethylamine (0.971
mL, 6.98 mmol) and 5-fluoro-thiophene-2-carboxylic acid (0.749 g,
5.12 mmol) was stirred overnight at room temperature. The solution
was poured into 2.5% NaHCO.sub.3 solution and the solids formed
were filtered, washed by cold water and air dried. The crude
product was purified by flash chromatography eluting with 0-10%
methanol in CH.sub.2Cl.sub.2 gradient to yield 2.16 g (83%) of
4-(3-fluoro-benzyl)-7-[4-(5-fluoro-thiophene-2-carbonyl)-piperazin-1-yl]--
5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid isopropyl
ester. .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.29 (d, J=6.0 Hz, 6H),
3.37 (m, 4H), 3.79 (m, 4H), 5.10 (m, 1H), 5.38 (s, 2H), 6.80 (m,
1H), 7.07 (m, 3H), 7.28 (t, J=3.6 Hz, 1H), 7.32 (d, J=5.6 Hz, 1H),
7.37 (m, 1H), 8.02 (d, J=5.6 Hz, 1H); EIMS m/z 558 (M+1).
Synthesis of
4-(3-fluoro-benzyl)-7-[4-(5-fluoro-thiophene-2-carbonyl)-piperazin-1-yl]--
5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl
ester (22)
[0161] This compound was prepared from
4-(3-fluoro-benzyl)-5-oxo-7-piperazin-1-yl-4,5-dihydro-thieno[3,2-b]pyrid-
ine-6-carboxylic acid ethyl ester (19) by applying the same
procedure as described for
4-(3-fluoro-benzyl)-7-[4-(5-fluoro-thiophene-2-carbonyl)-piperazin-1-yl]--
5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid isopropyl
ester (21). Yield 2.22 g (85%). .sup.1H-NMR (DMSO-d.sub.6) .delta.
1.29 (t, J=7.2 Hz, 3H), 3.39 (m, 4H), 3.82 (m, 4H), 4.27 (q, J=7.2
Hz, 2H), 5.38 (s, 2H), 6.80 (m, 1H), 7.10 (m, 3H), 7.29 (t, J=3.6
Hz, 1H), 7.33 (d, J=5.6 Hz, 1H), 7.36 (m, 1H), 8.03 (d, J=5.6 Hz,
1H); EIMS m/z 544 (M+1). ##STR136##
Alternative Method for the Synthesis of
4-(3-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-
-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester
(9)
[0162] A solution of
7-chloro-4-(3-fluoro-benzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-ca-
rboxylic acid ethyl ester (17) (45 g, 123 mmol),
1,4-diazabicyclo[2.2.2]octane (15.9 g, 141 mmol) and
piperazin-1-yl-thiophene-2-yl-methanone (27.8 g, 141 mmol) in dry
DMF was heated at 110.degree. C. for 7 h under argon. The solution
was cooled and poured into 2% ammonium chloride solution. The
solids formed were filtered and washed by cold water. The solids
were dissolved in dichloromethane and washed by water. The organic
phase was dried over MgSO.sub.4 and concentrated under vacuum to
yield 58.6 g (91%) of
4-(3-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-
-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester.
.sup.1H-NMR (DMSO-d.sub.6): .delta. 1.29 (t, J=7.2 Hz, 3H), 3.37
(m, 4H), 3.82 (m, 4H), 4.27 (q, J=7.2 Hz, 2H), 5.38 (s, 2H),
7.05-7.16 (m, 4H), 7.33 (d, J=5.6 Hz, 1H), 7.36 (m, 1H), 7.48 (dd,
J=1.2, 3.6 Hz, 1H), 7.79 (dd, J=1.2, 5.2 Hz, 1H), 8.03 (d, J=5.6
Hz, 1H); EIMS m/z 526 (M+1). Anal.
(C.sub.26H.sub.24FN.sub.3O.sub.4S.sub.2) C, H, N.
Synthesis of
4-(3-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-
-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid isopropyl ester
(23)
[0163] This compound was prepared from
7-chloro-4-(3-fluoro-benzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-ca-
rboxylic acid isopropyl ester (18) by applying the same procedure
as described for
4-(3-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-
-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid ethyl ester (9).
Yield (57%). .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.29 (d, J=6.4 Hz,
6H), 3.37 (m, 4H), 3.81 (m, 4H), 5.10 (m, 1H), 5.38 (s, 2H),
7.04-7.16 (m, 4H), 7.32 (d, J=5.6 Hz, 1H), 7.37 (m, 1H), 7.47 (dd,
J=1.2, 3.6 Hz, 1H), 7.79 (dd, J=1.2, 5.2 Hz, 1H), 8.02 (d, J=5.6
Hz, 1H); EIMS m/z 540 (M+1). ##STR137##
Synthesis of 3-(2-cyano-acetylamino)-thiophene-2-carboxylic acid
methyl ester (24)
[0164] A solution of methyl-3-amino-thiophene-2-carboxylate (13 g,
82.70 mmol) in neat methyl cyanoacetate (40 mL) was heated at
210.degree. C. for 10 h. The solution was cooled and excess methyl
cyanoacetate was distilled off under vacuum. The residue was taken
in methanol, sonicated briefly, and filtered. The solids were
washed by cold methanol and dried to yield 9.3 g (50%) of
3-(2-cyano-acetylamino)-thiophene-2-carboxylic acid methyl ester as
white solids. MP 146.degree. C.; .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 4.18 (s, 2H), 7.86 (d, J=5.2 Hz, 1H), 7.93 (d, J=5.2 Hz,
1H), 10.25 (s, 1H) ppm; MS m/z=225 amu (M.sup.++1). ##STR138##
Synthesis of
7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile
(25)
[0165] Sodium ethoxide (3.10 g, 45.62 mmol) was added to a solution
of 3-(2-cyano-acetylamino)-thiophene-2-carboxylic acid methyl ester
(24) (3.10 g, 45.62 mmol) in anhydrous ethanol and refluxed
overnight. The solution was cooled and excess solvent was distilled
off. The residue was dissolved in water and acidified to pH 2 by
cold diluted HCl solution. The solids formed were filtered, washed
by cold water, and dried under vacuum at room temperature to yield
6.7 g (84%) of
7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile as
white solids. MP>300.degree. C.; .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 6.96 (d, J=5.2 Hz, 1H), 8.04 (d, J=5.2 Hz, 1H), 12.10 (s,
1H) ppm; MS m/z=193 amu (M.sup.++1).
Synthesis of
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile
(26)
[0166] Triethylamine (11.78 mL, 84.54 mmol) was added to a solution
of 7-hydroxy-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile
(25) (6.5 g, 33.81 mmol) in neat phosphorous oxychloride at room
temperature and heated at 70.degree. C. for 1 h. The solution was
cooled and excess phosphorus oxychloride was distilled under
vacuum. The residue was suspended in water, and basified by solid
NaHCO.sub.3. The solids formed were filtered, washed by water, and
dried. The crude product was suspended in dichloromethane,
sonicated briefly, and filtered to yield
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile (9)
as white solids. Yield 5.3 g, (74%); MP 342.degree. C.; .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 7.11 (d, J=5.6 Hz, 1H), 8.29 (d, J=5.6
Hz, 1H) ppm; MS m/z=211 amu(M.sup.++1). ##STR139##
Synthesis of
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno3,2-b-
]pyridine-6-carbonitrile (27)
[0167] 1,4-Diazabicyclo[2.2.2]octane (3.19 g, 28.48 mmol) was added
to a solution of
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile
(26) (3.0 g, 14.24 mmol) and
piperazin-1-yl-thiophene-2-yl-methanone (3.07 g, 15.60 mmol) in dry
DMA. The solution was heated at 110.degree. C. for 1 h. The
solution was cooled and poured into ice water. The solids formed
were filtered, washed by water, and dried to yield 5-oxo-7-
[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno[3,2-b]pyridi-
ne-6-carbonitrile (4.72 g, 89%) as white solids. MP 293.degree. C.;
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 3.89 (m, 8H), 7.02 (d, J=5.6
Hz, 1H), 7.20 (dd, J=3.6, 5.2 Hz, 1H), 7.54 (dd, J=1.2 3.6 Hz, 1H),
7.84 (dd, J=1.2, 5.2 Hz, 1H), 8.16 (d, J=5.2 Hz, 1H) ppm; MS
m/z=371 amu (M.sup.++1).
General Procedures for Alkylation at nitrogen of Thienopyridinone
Moiety
[0168] The compounds referred to as compounds (28) through (31)
were prepared by applying either general procedure A or general
procedure B.
General Procedure A
[0169] A solution of
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno[3,2--
b]pyridine-6-carbonitrile (27) (1.35 mmol) in dry DMF was added to
a suspension of NaH (60% in mineral oil, 1.48 mmol) at room
temperature. The solution was stirred at room temperature for 15
min under argon. A corresponding alkyl halide (1.48 mmol) was added
to the solution and further stirred at room temperature until the
reaction was completed (TLC and/or LC-MS controlled). The solution
was poured into ice water and the solids formed were filtered,
washed by cold water, and dried. The crude product was purified by
flash chromatography eluting with 0-2% methanol in CH.sub.2Cl.sub.2
gradient to yield the compounds of preferred embodiments.
General Procedure B
[0170] A solution of
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-thieno[3,2--
b]pyridine-6-carbonitrile (27) (1.35 mmol), a corresponding alkyl
halide (2.70 mmol), and anhydrous potassium carbonate or cesium
carbonate (6.75 mmol) in DMF was heated overnight at 90.degree. C.
The solution was cooled and the solvent was distilled under reduced
pressure. The residue was suspended in water, sonicated briefly,
and filtered. The crude product was purified by flash
chromatography, eluting with 0-2% methanol in dichloromethane
gradient.
Synthesis of
4-(4-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-
-dihydro-thieno[3,2-b]pyridine-6-carbonitrile (28)
[0171] The compound was prepared by using 4-fluorobenzyl bromide
according to general procedure A to yield 260 mg (40%) white
solids. MP 286.degree. C.; 1H NMR (400 MHz, DMSO-d6) .delta. 3.88
(m, 8H), 5.36 (s, 2H), 7.13-7.18 (m, 3H), 7.30-7.35 (m, 3H), 7.51
(dd, J=1.2, 3.6 Hz, 1H), 7.81 (dd, J=1.2, 5.2 Hz, 1H), 8.21 (d,
J=5.2 Hz, 1H) ppm; MS m/z=479 amu (M.sup.++1). Anal.
(C.sub.24H.sub.19FN.sub.4O.sub.2S.sub.2) C, H, N.
Synthesis of
4-(3-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-
-dihydro-thieno[3,2-b]pyridine-6-carbonitrile (29)
[0172] The compound was prepared by using 3-fluorobenzyl bromide
according to general procedure A to yield 372 mg (56%) white
solids. MP 271.degree. C.; .sup.1H NMR (400 MHz, DMSO-d6) .delta.
3.89 (m, 8H), 5.39 (s, 2H), 7.05-7.13 (m, 3H), 7.16 (dd, J=3.6, 4.8
Hz, 1H), 7.31-7.37 (m, 2H), 7.51 (dd, J=1.2, 3.6 Hz, 1H), 7.81 (dd,
J=1.2, 5.2 Hz, 1H), 8.21 (d, J=5.2 Hz, 1H) ppm; MS m/z=479 amu
(M.sup.++1). Anal. (C.sub.24H.sub.19FN.sub.4O.sub.2S.sub.2) C, H,
N.
Synthesis of
5-oxo-4-(2-oxo-2-phenyl-ethyl)-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl-
]-4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile (30)
[0173] The compound was prepared by using 2-bromoacetophenone
according to general procedure A to yield 362 mg (55%) white
solids. MP 300.degree. C.; .sup.1H NMR (400 MHz, DMSO-d6) .delta.
3.92 (m, 8H), 5.78 (s, 2H), 7.17 (dd, J=3.6, 5.2 Hz, 1H), 7.36 (d,
J=6.0 Hz, 1H), 7.52 (dd, J=1.2, 3.6 Hz, 1H), 7.61-7.63 (m, 2H),
7.75 (m, 1H), 7.81 (dd, J=1.2, 5.2 Hz, 1H), 8.09 (dd, J=1.6, 7.2
Hz, 2H), 8.21 (d, J=5.6 Hz, 1H) ppm; MS m/z=489 amu (M.sup.++1).
Anal. (C.sub.25H.sub.20N.sub.4O.sub.3S.sub.2) C, H, N.
Synthesis of
5-oxo-4-pyridine-3-yl-methyl-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]--
4,5-dihydro-thieno[3,2-b]pyridine-6-carbonitrile (31)
[0174] The compound was prepared by using 3-bromomethyl pyridine
hydrobromide according to general procedure B to yield 310 mg (49%)
white solids. MP 247.degree. C.; .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 3.89 (m, 8H), 5.41 (s, 2H), 7.16 (dd, J=4.0, 5.2 Hz, 1H),
7.33 (dd, J=4.4, 7.6 Hz, 1H), 7.42 (d, J=5.6 Hz, 1H), 7.51 (d,
J=3.6 Hz, 1H), 7.61 (d, J=8.0 Hz, 1H), 7.81 (d, J=5.2 Hz, 1H), 8.23
(d, J=5.6 Hz, 1H), 8.47 (dd, J=1.6, 5.6 Hz, 1H), 8.57 (d, J=2.0 Hz,
1H) ppm; MS m/z=462 amu (M.sup.++1). Anal.
(C.sub.23H.sub.19N.sub.5O.sub.2S.sub.2) C, H, N.
Synthesis of 3-methoxycarbonylmethylsulfanyl-propionic acid methyl
ester (32)
[0175] Methyl acrylate (99.16 mL, 1.1 mol) was added slowly to a
solution of methyl thioglycolate (91 mL, 1 mol) and piperidine (2
mL) while maintaining the temperature of reaction mixture at
50.degree. C. The reaction mixture was stirred at 50.degree. C. for
2 h. Excess methyl acrylate and piperidine were distilled off under
high vacuum to yield 192 g (99%) of
3-methoxycarbonylmethylsulfanypropionic acid methyl ester as a
colorless viscous oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
2.63 (d, J=7.2 Hz, 2H), 2.89 (d, J=7.2 Hz, 2H), 3.24 (s, 2H), 3.68
(s, 3H), 3.72 (s, 3H) ppm.
Synthesis of 4-oxo-tetrahydro-thiophene-3-carboxylic acid methyl
ester (33)
[0176] A solution of 3-methoxycarbonylmethylsulfanypropionic acid
methyl ester (32) (58 g, 300 mmol) in dry THF (800 mL) was added
slowly within 4 h to a refluxing solution of hexane washed NaH (60%
in mineral oil, 13.24 g, 331 mmol) in THF. The solution was further
refluxed for 5 h. The solution was cooled and the solvent was
evaporated. The residue was taken in water, acidified to pH.about.1
by cold HCl solution, and extracted by CH.sub.2Cl.sub.2. The
combined organic phase was dried over MgSO.sub.4 and concentrated
under vacuum to get a viscous residue. The residue was purified by
flash chromatography eluting with hexane to yield 17 g (35%) of
4-oxo-tetrahydro-thiophene-3-carboxylic acid methyl ester as
colorless viscous oil which solidified on keeping overnight under
vacuum. MP 51.degree. C.; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
3.17-3.82 (m, 6.5 H), 10.94 (s, 0.5H) ppm. The isomeric mixture was
used to next step without further purification. ##STR140##
Synthesis of 4-hydroxyimino-tetrahydro-thiophene-3-carboxylic acid
methyl ester (34)
[0177] A suspension of 4-oxo-tetrahydro-thiophene-3-carboxylic acid
methyl ester (33) (16.96 g, 106 mmol), hydroxylamine hydrochloride
(16.96 g, 244 mmol) and barium carbonate (48.16 g, 244 mmol) in
methanol (800 mL) was refluxed overnight. The solution was cooled
and filtered. The filtrate was concentrated in vacuo. The residue
was suspended in water and extracted by ethyl acetate. The combined
organic phase was dried over MgSO.sub.4 and concentrated to yield
18.30 g (98%) of an isomeric mixture of
4-hydroxyimino-tetrahydro-thiophene-3-carboxylic acid methyl ester
as a viscous oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
3.14-4.11 (m, 7H), 8.17 (m, 1H) ppm. The isomeric mixture was used
in the following step without further purification.
Synthesis of 4-amino-thiophene-3-carboxylic acid methyl ester
(35)
[0178] HCl (1M solution in ether, 125 mL) was added slowly to a
solution of 4-hydroxyimino-tetrahydro-thiophene-3-carboxylic acid
methyl ester (34) (18.30 g, 104 mmol) in dry ether (200 mL) and dry
methanol (50 mL) stirred at room temperature. The solution was
further stirred at room temperature under argon for 24 h. The
solids formed were filtered, washed by cold ether, and dried to
yield 18.20 g (91%) of 4-amino-thiophene-3-carboxylic acid methyl
ester as the hydrochloride salt. MP 198.degree. C.; .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 3.86 (s, 3H), 7.22 (d, J=2.4 Hz, 1H),
8.37 (d, J=3.2 Hz, 1H) ppm; MS m/z=158 amu (M.sup.++1).
##STR141##
Synthesis of
4-(2-ethoxycarbonyl-acetylamino)-thiophene-3-carboxylic acid methyl
ester (36)
[0179] Pyridine (6.45 mL, 79.79 mmol) was added to a stirred
suspension of 4-amino-thiophene-3-carboxylic acid methyl ester
hydrochloride (35) (7 g, 36.27 mmol) in dry toluene at room
temperature. The solution was further stirred at room temperature
for 5 min and then cooled to -10+ C. Ethylmalonyl chloride (4.59
mL, 36.27 mmol) was added to this solution and further stirred at
-10.degree. C. for 1 h. The solution was allowed to come to room
temperature and poured into ice water. The product was extracted by
ethyl acetate. The combined organic phase was sequentially washed
by diluted HCl solution, saturated NaHCO.sub.3 solution, water, and
brine. The organic phase was dried over MgSO.sub.4 and concentrated
to yield 8.57 g (63%) of
4-(2-ethoxycarbonyl-acetylamino)-thiophene-3-carboxylic acid methyl
ester as a yellow viscous oil. .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 1.19 (t, J=7.2 Hz, 3H), 3.63 (s, 2H), 3.86 (s, 3H), 4.15
(q, J=7.2 Hz, 2H), 7.96 (d, J=3.6 Hz, 1H), 8.37 (d, J=3.6 Hz, 1H),
10.37 (s, 1H) ppm; MS m/z=240 amu (M.sup.++1).
Synthesis of
7-hydroxy-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
ethyl ester (37)
[0180] Sodium ethoxide (4.75 g, 66.34 mmol) was added to a solution
of 4-(2-ethoxycarbonyl-acetylamino)-thiophene-3-carboxylic acid
methyl ester (19) (8.57 g, 31.59 mmol) in anhydrous ethanol and
refluxed overnight. The solution was cooled and excess solvent was
distilled off. The residue was dissolved in water and acidified by
cold diluted HCl solution. The solids formed were filtered, washed
by water and dried under vacuum at room temperature to get 4.3 g
(57%) of
7-hydroxy-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
ethyl ester as white solid. MP 193.degree. C.; .sup.1H NMR (400
MHz, DMSO-d6) .delta. 1.27 (t, J=7.2 Hz, 3H), 4.28 (q, J=7.2 Hz,
2H), 6.83 (d, J=3.2 Hz, 1H), 8.24 (d, J=3.6 Hz, 1H), 11.10 (s, 1H),
12.90 (s, 1H) ppm; MS m/z=240 amu (M.sup.++1). ##STR142##
Synthesis of 5,7-dichloro-2-thia-4-aza-indene-6-carboxylic acid
ethyl ester (38)
[0181] A solution of
7-hydroxy-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
ethyl ester (37) (3.9 g, 16.30 mmol) in neat phosphorous
oxychloride (40 mL) was heated at 90.degree. C. for 4 h. The
solution was cooled and excess phosphorus oxychloride was distilled
under vacuum. The residue was suspended in water, sonicated
briefly, and filtered. The solids were dissolved in
CH.sub.2Cl.sub.2 and washed sequentially by saturated NaHCO.sub.3
solution, water, and brine. The organic phase was dried over
MgSO.sub.4 and concentrated under vacuum. The crude product was
purified by flash chromatography, eluting with CH.sub.2Cl.sub.2, to
yield 5,7-dichloro-2-thia-4-aza-indene-6-carboxylic acid ethyl
ester as white solids. Yield 2.1 g (46%); MP 79.degree. C.; .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 1.35 (t, J=7.2 Hz, 3H), 4.46 (q,
J=7.2 Hz, 2H), 8.47 (d, J=3.6 Hz, 1H), 8.53 (d, J=3.2 Hz, 1H) ppm;
MS m/z=276 amu (M.sup.+). ##STR143##
Synthesis of
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
ethyl ester (39)
[0182] Ammonium acetate (0.644 mg, 8.3 mmol) was added to a stirred
solution of 5,7-dichloro-2-thia-4-aza-indene-6-carboxylic acid
ethyl ester (38) (2.1 g, 7.6 mmol) in glacial acetic acid at room
temperature. The solution was heated at 140.degree. C. for 6 h. The
hot solution was poured over ice. The solids formed were filtered,
washed by water, and dried to yield 1.9 g (97%) of
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
ethyl ester as white solids. MP 157.degree. C.; .sup.1H NMR (400
MHz, DMSO-d6) .delta. 1.28 (t, J=7.2 Hz, 3H), 4.32 (q, J=7.2 Hz,
2H), 7.06 (d, J=3.6 Hz, 1H), 8.20 (d, J=3.2 Hz, 1H) ppm; MS m/z=258
amu (M.sup.++1).
Synthesis of
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4.5-dihydro-2-thia-4-az-
a-indene-6-carboxylic acid ethyl ester (40)
[0183] 1,4-Diazabicyclo[2.2.2]octane (2.07 g, 10.16 mmol) was added
to a solution of
7-chloro-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
ethyl ester (39) (2.4 g, 9.24 mmol) and
piperazin-1-yl-thiophene-2-yl-methanone (1.99 g, 10.16 mmol) in dry
DMA. The solution was heated at 110.degree. C. for 3 h. The
solution was cooled and the solvent was distilled. The residue was
suspended in water, sonicated briefly, and filtered. The solids
were washed by excess water and dried to yield
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2-thia-4-az-
a-indene-6-carboxylic acid ethyl ester (3.47 g, 90%) as white
solids. MP 282.degree. C.; .sup.1H NMR (400 MHz, DMSO-d6) .delta.
1.27 (t, J=7.2 Hz, 3H), 3.27 (m, 4H), 3.77 (m, 4H), 4.24 (q, J=7.2
Hz, 2H), 6.86 (d, J=3.6 Hz, 1H), 7.15 (dd, J=3.6, 5.2 Hz, 1H), 7.46
(dd, J=1.2, 3.6 Hz, 1H), 7.80 (dd, J=1.2, 4.8 Hz, 1H), 8.02 (d,
J=3.2 Hz, 1H), 11.30 (s, 1H) ppm; MS m/z=418 amu (M.sup.++1).
##STR144##
Synthesis of
5-oxo-4-(2-oxo-2-phenyl-ethyl)-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl-
]-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid ethyl ester
(41)
[0184] The compound was prepared from
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2-thia-4-az-
a-indene-6-carboxylic acid ethyl ester (40) and 2-bromoacetophenone
by applying general procedure A. Yield 66%; .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 1.25 (t, J=7.2 Hz, 3H), 3.34 (m, 4H), 3.86 (m,
4H), 4.24 (q, J=7.2 Hz, 2H), 5.58 (s, 2H), 7.15 (dd, J=3.6, 4.6 Hz,
1H), 7.24 (d, J=3.2 Hz, 1H), 7.48 (dd, J=1.2, 3.6 Hz, 1H), 7.60 (m,
2H), 7.71 (m, 1H), 7.80 (dd, J=1.2, 4.8 Hz, 1H), 8.09 (m, 2H), 8.16
(d, J=3.2 Hz, 1H) ppm; MS m/z=535 amu (M.sup.++1). Anal.
(C.sub.27H.sub.25N.sub.3O.sub.5N.
Synthesis of
4-(4-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-
-dihydro-2-thia-4-aza-indene-6-carboxylic acid ethyl ester (42)
[0185] The compound was prepared from
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2-thia-4-az-
a-indene-6-carboxylic acid ethyl ester (40) and
4-fluorobenzylbromide by applying general procedure A. Yield 54%;
MP 227.degree. C. .sup.1H NMR (400 MHz, DMSO-d6) .delta. 1.27 (t,
J=7.2 Hz, 3H), 3.30 (m, 4H), 3.83 (m, 4H), 4.27 (q, J=7.2 Hz, 2H),
5.18 (s, 2H), 7.14-7.17 (m, 3H), 7.20 (d, J=3.2 Hz, 1H), 7.33 (m,
2H), 7.46 (dd, J=1.2, 4.8 Hz, 1H), 7.80 (dd, J=1.2, 4.8 Hz, 1H),
8.13 (d, J=3.2 Hz, 1H) ppm; MS m/z=526 amu (M.sup.++1). Anal.
(C.sub.26H.sub.24FN.sub.3O.sub.4S.sub.2) C, H, N.
Synthesis of
4-(4-methoxycarbonyl-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin--
1-yl]-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid ethyl ester
(43)
[0186] The compound was prepared from
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2-thia-4-az-
a-indene-6-carboxylic acid ethyl ester (40) and
methyl-4-bromomethylbenzoate by applying general procedure A. Yield
51%; .sup.1H NMR (400 MHz, DMSO-d6) .delta. 1.25 (t, J=7.2 Hz, 3H),
3.30 (m, 4H), 3.82 (m, 7H), 4.27 (q, J=7.2 Hz, 2H), 5.28 (s, 2H),
7.12 (d, J=3.2 Hz, 1H), 7.16 (dd, J=5.2, 3.2 Hz, 1H), 7.37 (m, 2H),
7.47 (dd, J=1.2, 3.6 Hz, 1H), 7.79 (dd, J=1.2, 5.2 Hz, 1H), 7.91
(m, 2H), 8.13 (d, J=3.2 Hz, 1H) ppm; MS m/z=565 amu (M.sup.++1).
Anal. (C.sub.28H.sub.27N.sub.3O.sub.6S.sub.2)
Synthesis of
7-hydroxy-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
cyclohexylamide (44)
[0187] Cyclohexylamine (4.6 mL, 40.12 mmol) was added to a solution
of 7-hydroxy-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic
acid ethyl ester (39) (3.2 g, 82.70 mmol) in toluene and heated at
130.degree. C. for 4 h. The solution was cooled and excess solvent
was distilled off under vacuum. The residue was taken in
CH.sub.2Cl.sub.2, sonicated briefly, and filtered to yield 3.5 g
(89%) of
7-hydroxy-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
cyclohexylamide as white solids. MP 244.degree. C.; .sup.1H NMR
(400 MHz, DMSO-d6) .delta. 1.26-1.41 (m, 5H), 1.53 (m, 1H), 1.65
(m, 2H), 1.86 (m, 2H), 3.81 (m, 1H), 6.95 (s, 1H), 8.25 (s, 1H)
ppm; MS m/z=293 amu (M.sup.++1).
Synthesis of 5,7-dichloro-2-thia-4-aza-indene-6-carbonitrile
(45)
[0188] A solution of
7-hydroxy-5-oxo-4,5-dihydro-2-thia-4-aza-indene-6-carboxylic acid
cyclohexylamide (44) (3.5 g, 11.97 mmol) in neat phosphorous
oxychloride at room temperature was heated at 90.degree. C. for 3
h. The solution was cooled and excess phosphorus oxychloride was
distilled under vacuum. The residue was suspended in water, and
basified by solid NaHCO.sub.3. The solids formed were filtered
washed by water and dried to yield
5,7-dichloro-2-thia-4-aza-indene-6-carbonitrile as white solids.
Yield 2.5 g (91%); MP 228.degree. C.; .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 8.50 (m, 1H), 8.75 (m, 1H) ppm; MS m/z=229 amu
(M.sup.+). ##STR145##
Synthesis of
7-chloro-4,5-dihydro-5-oxo-2-thia-5-aza-indene-6-carbonitrile
(46)
[0189] Ammonium acetate (0.92 g, 12.0 mmol) was added to a stirred
solution of 5,7-dichloro-2-thia-4-aza-indene-6-carbonitrile (45)
(2.5 g, 10.91 mmol) in glacial acetic acid at room temperature. The
solution was heated at 140.degree. C. for 2 h. The hot solution was
poured over ice. The solids formed were filtered, washed by water,
and dried to yield 2.0 g (87%) of
7-chloro-4,5-dihydro-5-oxo-2-thia-5-aza-indene-6-carbonitrile as
white solids. MP 310.degree. C.; .sup.1H NMR (400 MHz, DMSO-d6)
.delta. 7.11 (d, J=3.2 Hz, 1H), 8.45 (d, J=3.2 Hz, 1H) ppm; MS
m/z=211 amu (M.sup.+). ##STR146##
Synthesis of
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2-thia-4-az-
a-indene-6-carbonitrile (47)
[0190] 1,4-Diazabicyclo[2.2.2]octane (2.13 g, 19.0 mmol) was added
to a solution of
7-chloro-4,5-dihydro-5-oxo-2-thia-5-aza-indene-6-carbonitrile (46)
(2.0 g, 9.5 mmol) and piperazin-1-yl-thiophene-2-yl-methanone (2.23
g, 11.4 mmol) in dry DMA. The solution was heated at 110.degree. C.
for 4 h. The solution was cooled and excess solvent was distilled
under vacuum. The residue was suspended in water, sonicated
briefly, filtered, washed by water, and dried to yield
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2-thia-4-az-
a-indene-6-carbonitrile (3.2 g, 91%) as white solids. MP
263.degree. C.; .sup.1H NMR (400 MHz, DMSO-d6) .delta. 3.89 (m,
8H), 6.90 (d, J=3.2 Hz, 1H), 7.17 (dd, J=3.6, 5.2 Hz, 1H), 7.50
(dd, J=1.2, 3.6 Hz, 1H), 7.81 (dd, J=1.2, 5.2 Hz, 1H), 8.28 (d,
J=3.2 Hz, 1H) ppm; MS m/z=371 amu (M.sup.++1).
[0191] The compounds referred to as compounds (48) through (51)
were prepared from
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2-thia-4-az-
a-indene-6-carbonitrile (47) by applying either general procedure A
or general procedure B described above.
Synthesis of
5-oxo-4-(2-oxo-2-phenyl-ethyl)-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl-
]-4,5-dihydro-2-thia-4-aza-indene-6-carbonitrile (48)
[0192] The compound was prepared by using 2-bromoacetophenone
according to general procedure A. Yield 56%. MP 308.degree. C.;
.sup.1H NMR (400 MHz, DMSO-d6) .delta. 3.95 (m, 8H), 5.60 (s, 2H),
7.17 (dd, J=3.6, 4.8 Hz, 1H), 7.27 (d, J=3.2 Hz, 1H), 7.52 (dd,
J=1.2, 3.6 Hz, 1H), 7.61-7.63 (m, 2H), 7.73 (m, 1H), 7.82 (dd,
J=1.2, 4.8 Hz, 1H), 8.09 (dd, J=1.2, 7.2 Hz, 2H), 8.43 (d, J=3.2
Hz, 1H) ppm; MS m/z=489 amu (M.sup.++1). Anal.
(C.sub.25H.sub.20N.sub.4O.sub.3S.sub.2) C, H, N.
Synthesis of
4-(4-fluoro-benzyl)-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-
-dihydro-2-thia-4-aza-indene-6-carbonitrile (49)
[0193] The compound was prepared by using 4-fluorobenzyl bromide
according to general procedure A. Yield 40%; MP 238.degree. C.;
.sup.1H NMR (400 MHz, DMSO-d 6) .delta. 3.92 (m, 8H), 5.19 (s, 2H),
7.15-7.19 (m, 4H), 7.30-7.35 (m, 2H), 7.50 (dd, J=1.2 3.6 Hz, 1H),
7.80 (dd, J=1.2, 5.2 Hz, 1H), 8.40 (d, J=3.2 Hz, 1H) ppm; MS
m/z=479 amu (M.sup.++1). Anal.
(C.sub.24H.sub.19FN.sub.4O.sub.2S.sub.2) C, H, N.
Synthesis of
4-{6-cyano-5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-5H-2-thia-4--
aza-indene-4-ylmethyl}-benzoic acid methyl ester (50)
[0194] The compound was prepared by using methyl 4-bromomethyl
benzoate according to general procedure A. Yield 56%; MP
263.degree. C.; .sup.1H NMR (400 MHz, DMSO-d 6) .delta. 3.82 (s,
3H), 3.93 (m, 8H), 5.29 (s, 2H), 7.12 (d, J=3.2 Hz, 1H), 7.17 (dd,
J=3.6, 4.8 Hz, 1H), 7.38 (d, J=8.0 Hz, 1H), 7.51 (dd, J=1.2, 3.6
Hz, 1H), 7.80 (dd, J=1.2, 5.2 Hz, 1H), 7.92 (d, J=8.0 Hz, 1H), 8.40
(d, J=2.8 Hz, 1H) ppm; MS m/z=519 amu (M.sup.++1). Anal.
(C.sub.26H.sub.22N.sub.4O.sub.4S.sub.2) C, H, N.
Synthesis of
5-oxo-4-pyridine-3-yl-methyl-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]--
4,5-dihydro-2-thia-4-aza-indene-6-carbonitrile (51)
[0195] The compound was prepared by using 3-chloromethyl pyridine
hydrochloride according to general procedure B. Yield 49%; MP
263.degree. C.; .sup.1H NMR (400 MHz, DMSO-d6) .delta. 3.92 (m,
8H), 5.25 (s, 2H), 7.17 (dd, J=4.0, 5.2 Hz, 1H), 7.28 (d, J=3.2 Hz,
1H), 7.33 (dd, J=4.8, 8.0 Hz, 1H), 7.50 (d, J=1.2 Hz, 1H), 7.66 (d,
J=1H), 7.80 (d, J=5.2 Hz, 1H), 8.40 (d, J=3.2 Hz, 1H), 8.45 (d,
J=4.8 Hz, 1H), 8.58 (s, 1H) ppm; MS m/z=462 amu (M.sup.++1). Anal.
(C.sub.23H.sub.19N.sub.5O.sub.2S.sub.2) C, H, N.
Synthesis of
5-oxo-4-pyridine-3-yl-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dih-
ydro-2-thia-4-aza-indene-6-carbonitrile (52)
[0196] Cu(OAc).sub.2 (363 mg, 2 mmol), pyridine-3-boronic acid (614
mg, 5 mmol), and triethylamine (560 .mu.L, 4 mmol) were added to a
solution of
5-oxo-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dihydro-2-thia-4-az-
a-indene-6-carbonitrile (47) (370 mg, 1 mmol) in wet DMF (DMF :
H.sub.2O :: 9: 1) stirred at room temperature. The solution was
further stirred at room temperature for 24 h. The solution was
passed through a bed of celite and the solvent was evaporated. The
residue was suspended in water and extracted by CH.sub.2Cl.sub.2.
The combined organic phase was partitioned with 10% aqueous HCl
solution. The aqueous phase was separated and the pH was adjusted
to 5 by 4 N NaOH solution. Then, a saturated solution of
NaHCO.sub.3 was added until the pH reached to 8. The solids formed
were filtered, washed by water, and dried under vacuum at
70.degree. C. to get
5-oxo-4-pyridine-3-yl-7-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-4,5-dih-
ydro-2-thia-4-aza-indene-6-carbonitrile as white solids. Yield 243
mg (54%); MP 282.degree. C.; .sup.1H NMR (400 MHz, DMSO-d6) .delta.
3.99 (m, 8H), 6.42 (d, J=2.8 Hz, 1H), 7.18 (dd, J=4.0, 4.8 Hz, 1H),
7.52 (d, J=2.8 Hz, 1H), 7.64 (dd, J=4.8, 8.0 Hz, 1H), 7.81 (d,
J=4.8 Hz, 1H), 7.88 (dm, 1H), 8.47 (d, J=3.2 Hz, 1H), 8.58 (d,
J=2.0 Hz, 1H), 8.71 (d, J=3.6 Hz, 1H) ppm; MS m/z=448 amu
(M.sup.++1). Anal. (C.sub.22HI.sub.7N.sub.5O.sub.2S.sub.2) C, H,
N.
Synthesis of methyl 3-(4-methoxybenzylamino)-thiophene-2-carboxylic
acid methyl ester (53 )
[0197] 4-Methoxybenzyl chloride (22.8 mL, 167.42 mmol) was added to
a solution of methyl-3-amino-thiophene-2-carboxylate (17.55 g, 111
mmol) in dry CH.sub.2Cl.sub.2 (20 mL). The solution was mixed well.
Excess CH.sub.2Cl.sub.2 was evaporated and the mixture was heated
overnight at 85.degree. C. under vacuum (3 torr). The mixture was
cooled to room temperature. Hexane was added to the mixture, which
was refluxed for 30 min, and cooled to 0.degree. C. The solids
formed were filtered, washed by hexane, and dried to yield 28.19 g
(91%) of 3-(4-methoxybenzylamino)-thiophene-2-carboxylic acid
methyl ester as red solids. .sup.1H NMR (400 MHz, DMSO-d6) .delta.
3.71 (s, 6H), 4.42 (s, 2H), 6.77 (d, J=5.6 Hz, 1H), 6.88 (d, J=8.8
Hz, 2H), 7.25 (d, J=8.8 Hz, 2H), 7.63 (d, J=5.6 Hz, 1H), ppm; MS
m/z=278 amu (M.sup.++1).
Synthesis of
7-hydroxy-4-(4-methoxybenzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-c-
arboxylic acid ethyl ester (54)
[0198] Solid NaH (60% in min. oil, 4.62 g, 135 mmol) was added in
portions to a stirred solution of
3-(4-methoxybenzylamino)thiophene-2-carboxylic acid methyl ester
(53) (26.69 g, 96 mmol) in dry DMF. The solution was stirred for 10
min. at room temperature and cooled to 0.degree. C. Ethyl malonyl
chloride was added slowly and the solution was further stirred at
room temperature for 15 min. Sodium ethoxide (13.1 g, 192 mmol) was
added to the solution and the solution was heated to 110.degree. C.
for 2 h. The solution was cooled and excess solvent was distilled
off. The residue was dissolved in a mixture of water (350 mL) and 4
M NaOH (50 mL), and the insoluble impurities were filtered off. The
filtrate was washed by diisopropyl ether and then acidified to pH 4
by cold diluted HCl solution. The solids formed were filtered,
washed by water, and dried under vacuum at room temperature to
yield 27 g (79%) of
7-hydroxy-4-(4-methoxybenzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-c-
arboxylic acid ethyl ester as off-white solids. .sup.1H NMR (400
MHz, DMSO-d6) .delta. 1.30 (t, J=7.2 Hz, 3H), 3.70 (s, 3H), 4.32
(q, J=7.2 Hz, 2H), 5.27 (s, 2H), 6.86 (d, J=8.8 Hz, 2H), 7.21 (d,
J=8.8 Hz, 2H), 7.34 (d, J=5.6 Hz, 1H), 8.14 (d, J=5.6 Hz, 1H),
13.31 (br, 1H) ppm; MS m/z=360 amu (M.sup.++1). ##STR147##
Synthesis of
7-chloro-4-(4-methoxybenzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-ca-
rboxylic acid ethyl ester (55)
[0199] Oxalyl chloride (4.8 mL, 55 mmol) was added to a solution of
7-hydroxy-4-(4-methoxybenzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-c-
arboxylic acid ethyl ester (54) (9.87 g, 27 mmol) in dry
CH.sub.2Cl.sub.2 at 0.degree. C. After adding anhydrous DMF (0.5
mL), the solution was allowed to come at room temperature and was
further stirred at room temperature for 24 h. Excess solvent was
evaporated to yield
7-chloro-4-(4-methoxybenzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-ca-
rboxylic acid ethyl ester as off white solids. Yield 85%; .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 1.30 (t, J=7.2 Hz, 3H), 3.70 (s,
3H), 5.36 (s, 2H), 6.88 (d, J=8.4 Hz, 2H), 7.24 (d, J=8.4 Hz, 2H),
7.53 (d, J=5.6 Hz, 1H), 8.20 (d, J=5.6 Hz, 1H ppm; MS m/z=378 amu
(M.sup.++H).
Synthesis of
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester (4)
[0200] A solution of
7-chloro-4-(4-methoxybenzyl)-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-ca-
rboxylic acid ethyl ester (55) (10.40 g, 27.5 mmol) in neat TFA was
heated at 70.degree. C. under argon for 36 h. The solution was
cooled and poured into ice water. The solids formed were filtered,
washed by water, and dried to yield 7.08 g (99%) of
7-chloro-5-oxo-4,5-dihydro-thieno[3,2-b]pyridine-6-carboxylic acid
ethyl ester as white solids. MP 206.degree. C.; .sup.1H NMR (400
MHz, DMSO-d6) .delta. 1.28 (t, J=7.2 Hz, 3H), 4.29 (q, J=7.2 Hz,
2H), 7.10 (d, J=5.2 Hz, 1H), 8.13 (d, J=5.2 Hz, 1H) ppm; MS m/z=240
amu (M.sup.++1). ##STR148##
Synthesis of 1H-thieno[2,3-d[1,3]oxazine-2,4-dione (56)
[0201] Methyl 2-amino-thiophene-3-carboxylate (5 g, 31.72 mmol) was
added to a solution of potassium hydroxide (3.55 g, 63.45 mmol) in
10 mL water. The solution was heated at 90.degree. C. until to get
a clear solution. The solution was then cooled to 0.degree. C. and
trichloromethyl chloroformate (5.74 mL, 47.57 mmol) was added
slowly. The solution was allowed to come to room temperature and
further stirred for 30 min. The precipitated solid was collected by
vacuum filtration to yield 4.7 g (88%) of
1H-thieno[2,3-d][1,3]oxazine-2,4-dione. MP 233.degree. C.
.sup.1H-NMR (DMSO-d.sub.6) .delta. 7.15 (d, J=6.0 Hz, 1H), 7.19 (d,
J=6.0 Hz, 1H) ppm.
Synthesis of 5-methyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione
(57)
[0202] Potassium hydroxide (14.77 g, 0.26 mol) was dissolved in 500
mL water. To this solution was added ethyl
2-amino-4-methyl-thiophene-3-carboxylate (24.38 g, 0.13 mol). The
solution was heated at 100.degree. C. for 16 hours. The solution
was then cooled to 0.degree. C. and trichloromethyl chloroformate
(23.8 mL, 0.20 mol) was added slowly. The solution was allowed to
come to room temperature and further stirred for 5 hours. The
precipitated solid was collected by vacuum filtration to yield 16.0
g (66%) of 5-methyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione. MP
220.degree. C. .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.30 (d, J=1.2
Hz, 3H), 6.78 (d, J=1.2 Hz, 1H), 12.55 (b, 1H) ppm; EIMS m/z 184
(M+1).
Synthesis of 1-benzyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione
(58)
[0203] NaH (60% dispersion in mineral oil, 1.0 g, 0.025 mol) was
added to a solution of 1H-thieno[2,3-d][1,3]oxazine-2,4-dione (56)
(3.53 g, 0.021 mol) in anhydrous DMF stirred at 0.degree. C. under
argon. The solution was stirred for 15 minutes before adding benzyl
bromide (2.97 mL, 0.025 mol). The solution was allowed to come to
room temperature and further stirred for 12 hours. The solution was
poured into ice water and the precipitated solid was collected by
vacuum filtration to yield 4.0 g (74%) of
1-benzyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 5.13 (s, 2H), 7.22 (d, J=5.6 Hz, 1H), 7.28
(d, J=5.6 Hz, 1H), 7.30-7.45 (m, 5H) ppm; EIMS m/z 260 (M+1).
##STR149##
Synthesis of
1-benzyl-5-methyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione (59)
[0204] This compound was prepared from
5-methyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione (57) by applying
the same method as described for the preparation of
1-benzyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione (58). Yield 4.2 g
(56%); MP 183.degree. C. .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.31
(s, 3H), 5.10 (s, 2H), 6.85 (s, 1H), 7.37 (m, 5H) ppm; EIMS m/z 274
(M+1).
Synthesis of
7-benzyl-4-hydroxy-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic
acid ethyl ester (60)
[0205] Diethyl malonate (2.14 mL, 0.014 mol) was added slowly to a
suspension of sodium hydride (60% dispersion in mineral oil, 0.66
g, 0.017 mol) in 50 mL anhydrous DMF stirred at 0.degree. C. under
argon. The solution was stirred for 15 minutes. Solid
1-benzyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione (58) (3.58 g, 0.014
mol) was added, and the solution was heated at 110.degree. C. for 2
hours. The solvent was then removed under vacuum, and the residue
was dissolved in water and washed with ethyl acetate. The aqueous
layer was acidified with dil. HCl to precipitate the product, which
was collected by vacuum filtration to give 3.09 g (68%) of
7-benzyl-4-hydroxy-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic
acid ethyl ester. MP 146.degree. C. .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.30 (t, J=7.2 Hz, 3H), 4.32 (q, J=7.2 Hz, 2H), 5.25 (s,
2H), 7.24-7.36 (m, 7H), 13.21 (b, 1H) ppm; EIMS m/z 330 (M+1).
Synthesis of
7-benzyl-4-hydroxy-3-methyl-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-car-
boxylic acid ethyl ester (61)
[0206] This compound was prepared from
1-benzyl-5-methyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione (59) by
applying the same method described for the preparation of
7-benzyl-4-hydroxy-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic
acid ethyl ester (60). Yield 80%; MP 166.degree. C. .sup.1H-NMR
(DMSO-d.sub.6): .delta. 1.32 (t, J=6.8 Hz, 3H), 2.40 (d, J=1.2 Hz,
3H), 4.36 (q, J=6.8 Hz, 2H), 5.22 (s, 2H), 6.84 (d, J=1.2 Hz, 1H),
7.25 (m, 3H), 7.32 (m, 2H), 14.04 (b, 1H) ppm; EIMS m/z 344 (M+1).
##STR150##
Synthesis of
7-benzyl-6-oxo-4-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-6,7-dihydro-th-
ieno[2,3-b]pyridine-5-carboxylic acid ethyl ester (62)
[0207] Oxalyl chloride (0.66 mL, 0.008 mol) was added slowly to a
solution of
7-benzyl-4-hydroxy-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic
acid ethyl ester (60) (1.0 g, 0.003 mol) in 25 mL anhydrous DMF
stirred at -45.degree. C. under argon. The solution was heated to
70.degree. C. for 4 hours, and then poured into water. A small
amount of brine was added, and the precipitated solid was collected
by vacuum filtration. The oily solid was dissolved in
dichloromethane, dried with magnesium sulfate, and then
concentrated under vacuum to yield an oil. The oil was dissolved in
DMF. Piperazin-1-yl-thiophen-2-yl-methanone (1.2 g, 0.004 mol) and
DABCO (0.68 g, 0.006 mol) were added to the solution under argon.
The solution was heated to 110.degree. C. for 5 hours, and then
poured into a 5% ammonium chloride aqueous solution. The
precipitated solid was collected by vacuum filtration, and then
purified by reverse phase (MeCN/water) chromatography to yield
0.320 g (21%) of
7-benzyl-6-oxo-4-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-6,7-dihydro-th-
ieno[2,3-b]pyridine-5-carboxylic acid ethyl ester. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.28 (t, J=7.2 Hz, 3H), 3.40 (b, 4H), 3.81
(b, 4H), 4.26 (q, J=7.2 Hz, 2H), 5.26 (s, 2H), 7.15 (m, 1H), 7.28
(m, 5H), 7.35 (m, 2H), 7.47 (m, 1H), 7.79 (m, 1H) ppm; EIMS m/z 508
(M+1).
Synthesis of
7-benzyl-3-methyl-6-oxo-4-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-6,7-d-
ihydro-thieno[2,3-b]pyridine-5-carboxylic acid ethyl ester (63)
[0208] This compound was prepared from
7-benzyl-4-hydroxy-3-methyl-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-car-
boxylic acid ethyl ester (61) by applying the method described for
the preparation of
7-benzyl-6-oxo-4-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-6,7-dihydro-th-
ieno[2,3-b]pyridine-5-carboxylic acid ethyl ester (62). Yield 16%;
MP 195.degree. C. .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.29 (t, J=7.2
Hz, 3H), 2.53 (d, J=1.2 Hz, 3H), 4.29 (q, J=7.2 Hz, 2H), 5.27 (s,
2H), 6.94 (d, J=1.2 Hz, 1H), 7.16 (m, 1H), 7.28 (m, 3H), 7.33 (m,
2H), 7.43 (dd, J=0.8, 3.6 Hz, 1H), 7.79 (dd, J=0.8, 4.8 Hz, 4.8 Hz,
1H), ppm; EIMS m/z 522 (M+1).
Synthesis of
7-benzyl-4-chloro-6,7-dihydro-6-oxo-thieno[2,3-b]pyridine-5-carbonitrile
(64)
[0209] Cyclohexylamine (1.44 mL, 0.013 mol) was added to a solution
of ethyl
7-benzyl-4-hydroxy-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-carbox-
ylic acid ethyl ester (60) (2.07 g, 0.006 mol) in toluene stirred
under argon. The solution was refluxed for 4 hours, and then the
toluene was removed under vacuum. The residue was dissolved in
dichloromethane and washed with sodium bisulfate solution. The
organic layer was dried over magnesium sulfate and concentrated
under vacuum to give an oil. The oil was dissolved in 25 mL
phosphorus oxychloride and cooled to 0.degree. C., and
triethylamine (2.18 mL, 0.016 mol) was added. The reaction was
heated to 100.degree. C. for 4 days. The solution was cooled and
excess phosphorus oxychloride was removed under vacuum. The residue
was suspended in water and the solid was collected by vacuum
filtration. The solid was dissolved in dichloromethane, washed with
saturated solution of sodium bicarbonate, water, and brine. The
organic layer was dried over magnesium sulfate and concentrated
under vacuum to give 1.56 g (83%) of
7-benzyl-4-chloro-6,7-dihydro-6-oxo-thieno[2,3-b]pyridine-5-carbonitrile.
.sup.1H-NMR (DMSO-d.sub.6) .delta. 5.37 (s, 2H), 7.34 (m, 6H), 7.49
(m, 1H) ppm; EIMS m/z 301 (M+1).
Synthesis of
7-benzyl-4-chloro-6,7-dihydro-3-methyl-6-oxo-thieno[2,3-b]pyridine-5-carb-
onitrile (65)
[0210] This compound was prepared from
7-benzyl-4-hydroxy-3-methyl-6-oxo-6,7-dihydro-thieno[2,3-b]pyridine-5-car-
boxylic acid ethyl ester (61) by applying a similar procedure as
described for the preparation of
7-benzyl-4-chloro-6,7-dihydro-6-oxo-thieno[2,3-b]pyridine-5-carbonitrile
(64). Yield 87%. .sup.1H-NMR (DMSO-d.sub.6): .delta. 2.48 (s, 3H),
5.36 (s, 2H), 7.14 (s, 1H), 7.32 (m, 5H) ppm; EIMS m/z 315 (M+1).
##STR151##
Synthesis of
7-benzyl-6-oxo-4-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-6,7-dihydro-th-
ieno[2.3-b]pyridine-5-carbonitrile (66)
[0211] A solution of
7-benzyl-4-chloro-6,7-dihydro-6-oxo-thieno[2,3-b]pyridine-5-carbonitrile
(64) (0.750 g, 0.0025 mol), piperazin-1-yl-thiophen-2-yl-methanone
(0.69 g, 0.0035 mol), and DABCO (0.56 g, 0.005 mol) in anhydrous
DMF was heated at 110.degree. C. for 12 h. The solution was cooled
and poured into a 5% ammonium chloride aqueous solution and the
precipitated solid was collected by vacuum filtration. The solid
was purified by reverse phase chromatography (MeCN/water) to yield
0.330 g (29%) of
7-benzyl-6-oxo-4-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-6,7-dihydro-th-
ieno[2,3-b]pyridine-5-carbonitrile. MP 257.degree. C.; 1H-NMR
(DMSO-d6) .delta. 3.83 (m, 8H), 5.28 (s, 2H), 7.16 (m, 1H), 7.33
(m, 7H), 7.51 (d, J=3.2 Hz, 1H), 7.81 (d, J=4.8 Hz, 1H) ppm; EIMS
m/z 461 (M+1).
Synthesis of
7-benzyl-3-methyl-6-oxo-4-r4-(thiophene-2-carbonyl)-piperazin-1-yl]-6,7-d-
ihydro-thieno[2,3-b]pyridine-5-carbonitrile (67)
[0212] This compound was prepared from
7-benzyl-4-chloro-6,7-dihydro-3-methyl-6-oxo-thieno[2,3-b]pyridine-5-carb-
onitrile (65) by applying the same method as described for the
preparation of
7-benzyl-6-oxo-4-[4-(thiophene-2-carbonyl)-piperazin-1-yl]-6,7-dihydro-
-thieno[2,3-b]pyridine-5-carbonitrile (66). Yield 25%; MP
223.degree. C. .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.48 (d, J=0.8
Hz, 3H), 3.50 (m, 4H), 3.86 (b, 4H), 5.30 (s, 2H), 6.99 (d, J=1.2
Hz, 1H), 7.16 (m, 1H), 7.30 (m, 5H), 7.47 (dd, J=0.8, 3.6 Hz, 1H),
7.80 (dd, J=1.2, 5.2 Hz, 1H) ppm; EIMS m/z 475 (M+1).
Synthesis of 6-methyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione
(68)
[0213] Methyl 2-amino-5-methyl-thiophene-3-carboxylate (16.8 g, 299
mmol) was added to a solution of potassium hydroxide (25 g, 146
mmol) in 300 mL water. The solution was heated at 90.degree. C.
until to yield a clear solution. The solution was heated at same
temperature for another 30 min and then cooled to 0.degree. C.
Trichloromethyl chloroformate (26.42 mL, 219 mmol) was added slowly
without allowing the temperature to rise beyond 10.degree. C. The
solution was further stirred for 2 h. The precipitated solid was
collected by vacuum filtration to yield 21.6 g (83%) of
6-methyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.38 (d, J=1.2 Hz, 3H), 6.90 (q, J=1.2 Hz,
1H), 12.44 (b, 1H) ppm; EIMS m/z 184 (M+1). ##STR152##
Synthesis of
7-(3-fluoro-benzyl)-4-hydroxy-2-methyl-6-oxo-6,7-dihydro-thieno[2,3-b]pyr-
idine-5-carboxylic acid ethyl ester (69)
[0214] Sodium hydride (60% dispersion in mineral oil, 5.6 g, 0.140
mol) was added slowly to a solution of
6-methyl-1H-thieno[2,3-d][1,3]oxazine-2,4-dione (68) (11.07 g,
0.060 mol) in anhydrous DMF stirred under argon at 0.degree. C. The
solution was stirred for 15 minutes before adding 3-fluoro
benzylbromide (7.6 mL, 0.062 mol). The solution was allowed to come
at room temperature and further stirred for 3 hours. The solution
was cooled to -10.degree. C. and diethylmalonate (9.36 mL, 0.061
mol) was added slowly. The solution was heated at 110.degree. C.
(TLC control). The reaction was cooled and poured into aqueous
sodium carbonate (7.7 g, 0.072 mol) solution. The aqueous solution
was washed with isopropyl ether and acidified to pH 2 with dil.
HCl. The solids were filtered, washed by cold water and air dried
to yield 16.9 g (97%) of
7-(3-fluoro-benzyl)-4-hydroxy-2-methyl-6-oxo-6,7-dihydro-thieno[2,3-b]pyr-
idine-5-carboxylic acid ethyl ester (69). .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.30 (t, J=7.2 Hz, 3H), 2.40 (d, J=1.2 Hz,
3H), 4.32 (q, J=7.2 Hz, 2H), 5.22 (s, 2H), 7.05-7.15 (m, 4H), 7.38
(m, 1H), 13.25 (s, 1H) ppm; EIMS m/z 362 (M+1).
Synthesis of
4-chloro-7-(3-fluoro-benzyl)-2-methyl-6-oxo-6,7-dihydro-thieno[2,3-b]pyri-
dine-5-carboxylic acid ethyl ester (70)
[0215] Oxalyl chloride (4.83 mL, 0.055 mol) was added to a solution
of
7-(3-fluoro-benzyl)-4-hydroxy-2-methyl-6-oxo-6,7-dihydro-thieno[2,3-b]pyr-
idine-5-carboxylic acid ethyl ester (69) (8 g, 0.022 mol) in DMF at
-30.degree. C. under argon. The temperature was raised gradually to
75.degree. C. and stirred for 3 h. The solution was cooled and
poured into ice water. The solids formed were filtered, washed by
cold water and dried to yield 6.8 g (82 %) of
4-chloro-7-(3-fluoro-benzyl)-2-methyl-6-oxo-6,7-dihydro-thieno[2,3-b]pyri-
dine-5-carboxylic acid ethyl ester. .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.30 (t, J=7.2 Hz, 3H), 2.45 (d, J=1.2 Hz, 3H), 4.33 (q,
J=7.2 Hz, 2H), 5.31 (s, 2H), 7.05 (d, J=1.2 Hz, 1H), 7.09-7.17 (m,
3H), 7.40 (m, 1H) ppm; EIMS m/z 380 (M+1).
Synthesis of
7-(3-fluoro-benzyl)-2-methyl-6-oxo-4-[4-(thiophene-2-carbonyl)-piperazin--
1-yl]-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic acid ethyl
ester (71)
[0216] A solution of
4-chloro-7-(3-fluoro-benzyl)-2-methyl-6-oxo-6,7-dihydro-thieno[2,3-b]pyri-
dine-5-carboxylic acid ethyl ester (70) (3.0 g, 7.89 mmol),
piperazin-1-yl-thiophen-2-yl-methanone (1.78 g, 9.09 mmol), and
DABCO (1.02 g, 9.09 mmol) in anhydrous DMF was heated overnight at
110.degree. C. The solution was cooled and poured into a 2%
ammonium chloride aqueous solution. The precipitated solid was
collected by vacuum filtration. The solid was redissolved in
dichloromethane and filtered. The filtrate was concentrated under
vacuum and the residue was purified by flash chromatography eluting
with 0-5% MeOH in CH.sub.2Cl.sub.2 gradient to yield 2.8 g (65%) of
7-(3-fluoro-benzyl)-2-methyl-6-oxo-4-[4-(thiophene-2-carbonyl)-piperazin--
1-yl]-6,7-dihydro-thieno[2,3-b]pyridine-5-carboxylic acid ethyl
ester. .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.29 (t, J=7.2 Hz, 3H),
2.43 (s, 3H), 3.29 (m, 4H), 3.81 (m, 4H), 4.26 (q, J=7.2 Hz, 2H),
5.23 (s, 2H), 6.99 (d, J=1.2 Hz, 1H), 7.06-7.16 (m, 4H), 7.41 (m,
1H), 7.46 (dd, J=1.2, 4.0 Hz, 1H), 7.79 (dd, J=1.2,4.8 Hz, 1H);
EIMS m/z 540(M+1). ##STR153## Antibody Binding Assay of MIF
Inhibitors
[0217] An antibody binding assay for MIF inhibitors was performed
on selected compounds in a 96-well format using MIF produced by
THP-1 cells. The THP-1 cells were washed once with 250 .mu.l of 0.1
M sodium carbonate, pH 11.4, for 1 to 2 minutes and immediately
aspirated. The THP-1 cells were then washed with media containing
0.5% FBS plus 25 .mu.g/ml heparin and then incubated in this same
medium for the indicated lengths of time. The THP-1 cells were
resuspended to approx. 5.times.10.sup.6 cells/ml in RPMI medium
containing 20 .mu.g/ml of bacterial LPS and incubated for 18-20
hours, after which THP-1 cell supernatant was collected and
incubated with a candidate compound. A 96-well ELISA plate (Costar
Number 3590) was coated with a MIF monoclonal antibody (R&D
Systems Catalog Number MAB289) at a concentration of 4 .mu.g/ml for
two hours at 37.degree. C. Undiluted THP-1 cell culture supernatant
incubated with the candidate compound was added to the ELISA plate
for a two-hour incubation at room temperature. The wells were then
washed, a biotinylated MIF polyclonal antibody (R&D Systems
#AF-289-PB) was added followed by Streptavidin-HRP and a
chromogenic substrate. The amount of MIF was calculated by
interpolation from an MIF standard curve. The in vitro MIF
IC.sub.50 (nM) of selected candidate compounds is provided in Table
1.
[0218] Each of the compounds tested exhibited MIF inhibiting
activity in the assay, i.e., they inhibit MIF activity and are
therefore suitable for use as pharmaceutical compositions for the
treatment of diseases mediated by MIF. Assay test results for
preferred compounds are provided in Table 1. TABLE-US-00001 TABLE 1
In vitro MIF IC.sub.50 Compound Structure (nM) 101 ##STR154## 70
102 ##STR155## 82 103 ##STR156## 437 104 ##STR157## 59 105
##STR158## 53 106 ##STR159## 40 107 ##STR160## 35 108 ##STR161##
171 109 ##STR162## 233 110 ##STR163## 1,734 111 ##STR164## 254 112
##STR165## 595 113 ##STR166## 419 114 ##STR167## 70 115 ##STR168##
11,800 116 ##STR169## 139 117 ##STR170## 12,260 119 ##STR171##
4,150 120 ##STR172## 799 121 ##STR173## 146 122 ##STR174## 5,840
123 ##STR175## 8,770 124 ##STR176## 269 125 ##STR177## 387 126
##STR178## 648 127 ##STR179## 32
[0219] Amongst other uses, the MIF inhibitors of the preferred
embodiments, including the specific compounds in Table 1, are
useful for the treatment of ed diseases, such as inflammatory and
autoimmune diseases including, but to arthritis, uveoritinitis,
colitis (including Crohm's disease and ulcerative hiritis, atopic
dermatitis, psoriasis, proliferative vascular disease,
cytokine-oxicity, sepsis, septic shock, interleukin-2 toxicity,
acute respiratory distress (ARDS), asthma, insulin-dependent
diabetes, multiple sclerosis, osis, graft versus host disease,
lupus syndromes, and other conditions ed by local or systemic
MIF-release or synthesis. The MIF inhibitors of the preferred
embodiments can, e.g., further be used to treat tumor growth and
angiogenesis, and to treat malaria. The minimum inhibitory
concentration (MIC) of the MIF inhibitors of Table 1, as well as
other MIF inhibitors of preferred embodiments as described herein,
against these diseases or conditions is from about 1.0 nM to about
100 .mu.M. For the treatment of these diseases or conditions, the
MIF inhibitors of the preferred embodiments can be administered to
larger mammals, for example humans, by oral, intravenous or
intra-muscular administration at doses of from less than about 1.0
mg to dosages of 100 mg/Kg or more, as are used with other
conventional therapies.
[0220] In diseases and conditions which are mediated by MIF, such
as those described herein, treatment comprises administering to a
subject in need of such treatment an effective amount of a MIF
inhibitor of Table 1, or one or more other MIF inhibitors of the
preferred embodiments in the form of a pharmaceutical composition.
The term "treatment" as used herein is a broad term, and is to be
given its ordinary and customary meaning to a person of ordinary
skill in the art (and is not to be limited to a special or
customized meaning), and refers without limitation to both
treatment of an existing disease or condition, as well as
prophylaxis. For such treatment, the appropriate dosage depends
upon, for example, the chemical nature, and the pharmacokinetic
data of the MIF inhibitor, the individual host, the mode of
administration and the nature and severity of the disease or
condition being treated. However, in general, for satisfactory
results in larger mammals, for example humans, an indicated daily
dosage of from about 0.01 g to about 1.0 g, of a MIF inhibitor of
the preferred embodiments can be conveniently administered, for
example, in a single dose or in divided doses up to two, three, or
four or more times a day.
[0221] Various MIF inhibitors and methods of preparing and using
the same, as well as assays for use in determining MIF inhibiting
activity of candidate compounds, are disclosed in U.S. Publication
No. US-2003-0195194-A1; U.S. Publication No. US-2004-0204586-A1;
and U.S. Publication No. US-2005-0124604-A1.
[0222] All references cited herein, including but not limited to
published and unpublished applications, patents, and literature
references, are incorporated herein by reference in their entirety
and are hereby made a part of this specification. To the extent
publications and patents or patent applications incorporated by
reference contradict the disclosure contained in the specification,
the specification is intended to supersede and/or take precedence
over any such contradictory material.
[0223] The term "comprising" as used herein is synonymous with
"including," "containing," or "characterized by," and is inclusive
or open-ended and does not exclude additional, unrecited elements
or method steps.
[0224] All numbers expressing quantities of ingredients, reaction
conditions, and so forth used in the specification are to be
understood as being modified in all instances by the term "about."
Accordingly, unless indicated to the contrary, the numerical
parameters set forth herein are approximations that may vary
depending upon the desired properties sought to be obtained. At the
very least, and not as an attempt to limit the application of the
doctrine of equivalents to the scope of any claims in any
application claiming priority to the present application, each
numerical parameter should be construed in light of the number of
significant digits and ordinary rounding approaches.
[0225] The above description discloses several methods and
materials of the present invention. This invention is susceptible
to modifications in the methods and materials, as well as
alterations in the fabrication methods and equipment. Such
modifications will become apparent to those skilled in the art from
a consideration of this disclosure or practice of the invention
disclosed herein. Consequently, it is not intended that this
invention be limited to the specific embodiments disclosed herein,
but that it cover all modifications and alternatives coming within
the true scope and spirit of the invention.
* * * * *
References