U.S. patent application number 12/628306 was filed with the patent office on 2010-11-04 for vasculostatic agents and methods of use thereof.
This patent application is currently assigned to TargeGen Inc.. Invention is credited to Elena Dneprovskaia, John Doukas, Xianchang Gong, John D. Hood, Glenn Noronha, Ivor Royston, Ute Splittgerber, Wolfgang Wrasidlo, Ningning Zhao.
Application Number | 20100278811 12/628306 |
Document ID | / |
Family ID | 44361236 |
Filed Date | 2010-11-04 |
United States Patent
Application |
20100278811 |
Kind Code |
A1 |
Wrasidlo; Wolfgang ; et
al. |
November 4, 2010 |
VASCULOSTATIC AGENTS AND METHODS OF USE THEREOF
Abstract
Compositions and methods and are provided for treating disorders
associated with compromised vasculostasis. Invention methods and
compositions are useful for treating a variety of disorders
including for example, stroke, myocardial infarction, cancer,
ischemia/reperfusion injury, autoimmune diseases such as rheumatoid
arthritis, eye diseases such as retinopathies or macular
degeneration or other vitreoretinal diseases, inflammatory
diseases, vascular leakage syndrome, edema, transplant rejection,
adult/acute respiratory distress syndrome (ARDS), and the like.
Inventors: |
Wrasidlo; Wolfgang; (La
Jolla, CA) ; Doukas; John; (Encinitas, CA) ;
Royston; Ivor; (La Jolla, CA) ; Noronha; Glenn;
(Oceanside, CA) ; Hood; John D.; (San Diego,
CA) ; Dneprovskaia; Elena; (San Diego, CA) ;
Gong; Xianchang; (La Jolla, CA) ; Splittgerber;
Ute; (San Francisco, CA) ; Zhao; Ningning;
(San Diego, CA) |
Correspondence
Address: |
GOODWIN PROCTER LLP;PATENT ADMINISTRATOR
53 STATE STREET, EXCHANGE PLACE
BOSTON
MA
02109-2881
US
|
Assignee: |
TargeGen Inc.
|
Family ID: |
44361236 |
Appl. No.: |
12/628306 |
Filed: |
December 1, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11653190 |
Jan 11, 2007 |
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12628306 |
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10679209 |
Oct 2, 2003 |
7208493 |
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11653190 |
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60479295 |
Jun 17, 2003 |
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60466983 |
Apr 30, 2003 |
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60463818 |
Apr 17, 2003 |
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60443752 |
Jan 29, 2003 |
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60440234 |
Jan 14, 2003 |
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60415981 |
Oct 3, 2002 |
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Current U.S.
Class: |
424/130.1 ;
424/649; 514/234.2; 514/243; 514/249; 514/266.31; 514/3.1; 514/34;
514/411; 514/415 |
Current CPC
Class: |
A61P 9/00 20180101; C07D
403/12 20130101; A61P 37/06 20180101; C07D 405/12 20130101; C07D
209/14 20130101; C07D 209/48 20130101; A61P 19/02 20180101; C07D
239/88 20130101; A61P 9/10 20180101; C07D 471/04 20130101; A61P
11/00 20180101; C07D 253/10 20130101; C07D 487/04 20130101; C07D
239/95 20130101; C07D 405/04 20130101; A61P 43/00 20180101; C07D
241/42 20130101; A61P 17/02 20180101; C07D 239/90 20130101; A61P
27/02 20180101; A61P 35/00 20180101; A61P 11/06 20180101; A61P
29/00 20180101; C07D 401/12 20130101 |
Class at
Publication: |
424/130.1 ;
514/415; 514/234.2; 514/249; 514/243; 514/266.31; 514/411; 514/3.1;
424/649; 514/34 |
International
Class: |
A61K 39/395 20060101
A61K039/395; A61K 31/404 20060101 A61K031/404; A61K 31/5377
20060101 A61K031/5377; A61K 31/519 20060101 A61K031/519; A61K 31/53
20060101 A61K031/53; A61K 31/517 20060101 A61K031/517; A61K 31/407
20060101 A61K031/407; A61K 38/14 20060101 A61K038/14; A61K 33/24
20060101 A61K033/24; A61K 31/704 20060101 A61K031/704; A61P 35/00
20060101 A61P035/00; A61P 11/06 20060101 A61P011/06; A61P 27/02
20060101 A61P027/02; A61P 9/00 20060101 A61P009/00 |
Claims
1. A method for treating a subject having, or at risk of
developing, a disorder or a disease selected from the group
consisting of compromised vasculostasis, myocardial infarction,
stroke, congestive heart failure, an ischemia or reperfusion
injury, cancer, arthritis or other arthropathy, retinopathy or
vitreoretinal disease, macular degeneration, autoimmune disease,
vascular leakage syndrome, an inflammatory disease, edema,
transplant rejection, burn, or acute or adult respiratory distress
syndrome (ARDS), comprising administering to a subject in need
thereof an effective amount of at least one compound having the
general structure III, or any combination thereof, or tautomers,
pharmaceutically acceptable salts, hydrates, solvates, crystal
forms and individual diastereomers thereof: ##STR00209## wherein:
each of Z.sub.1-Z.sub.6 is independently selected from a group
consisting of C, --C.dbd.O, N, and NR.sup.a, wherein R.sup.a is
selected from a group consisting of --H, an alkyl, or a substituted
alkyl, wherein said substituent is selected from a group consisting
of a halogen, hydroxy, oxo, and amino, each X is independently
selected from a group consisting of a halogen, --OR.sup.b,
--NR.sup.b.sub.2, and --SR.sup.b, wherein R.sup.b is selected from
a group consisting of --H, a lower alkyl,
--(CH.sub.2).sub.2NH(CH.sub.2CH.sub.3),
--(CH.sub.2).sub.3morpholyn-1-yl,
--(CH.sub.2).sub.3(N-methylpiperazinyn-1-yl), an aryl, a
heteroaryl, --(NH--NHR.sup.c), and --(N.dbd.N--NH--R.sup.c),
wherein R.sup.c is H or a lower alkyl, each Y is independently
selected from a group consisting of --OR.sup.d, --NR.sup.d.sub.2,
--SR.sup.d, and --OPO.sub.3H.sub.2, wherein R.sup.d is selected
from a group consisting of H, a lower alkyl, an aryl, a heteroaryl,
--(CH.sub.2).sub.2NH(CH.sub.2CH.sub.3),
--(CH.sub.2).sub.3morpholyn-1-yl, and --(CH.sub.2).sub.3
(N-methylpiperazinyn-1-yl); or each Y is independently selected
from a group consisting of an alkyl, a substituted alkyl, an aryl,
a substituted aryl, a heteroaryl, a substituted heteroaryl, and a
halogen, wherein said substituent is selected from a group
consisting of a halogen, --OR.sup.e, --NR.sup.e.sub.2, --SR.sup.e,
and --P(O)(OH).sub.2, wherein R.sup.e is selected from a group
consisting of H, a lower alkyl, an aryl, and a heteroaryl; or each
Y is independently selected from a group consisting of
CH.sub.2glycinyl, CH.sub.2NHethoxy, CH.sub.2NHCH.sub.2alkyl,
CH.sub.2NHCH.sub.2t-Bu, CH.sub.2NHCH.sub.2aryl,
CH.sub.2NHCH.sub.2substituted aryl, CH.sub.2NHCH.sub.2heteroaryl,
and CH.sub.2NHCH.sub.2-substituted heteroaryl; or when n is 2, each
Y is taken together to form a fused aromatic or heteroaromatic ring
system; and each of m and n is independently an integer having the
value between 1 and 4, wherein when each of Z.sub.1, Z.sub.3,
Z.sub.5, and Z.sub.6 is N, X is NH.sub.2, and m=n=2, Y is not
phenyl or 4-hydroxyphenyl, thereby treating the disorder.
2. The method of claim 1, wherein the compound III has the
structure: ##STR00210## wherein: each of m and n is independently
an integer having the value between 1 and 4, each Y is
independently selected from a group consisting of --OR.sup.d,
--NR.sup.d.sub.2, --SR.sup.d, and --OPO.sub.3H.sub.2, wherein
R.sup.d is selected from a group consisting of H, a lower alkyl, an
aryl, and --(CH.sub.2).sub.2NH(CH.sub.2CH.sub.3), or each Y is
independently selected from a group consisting of an alkyl, a
substituted alkyl, an aryl, a substituted aryl, and a halogen,
wherein said substituent is selected from a group consisting of a
halogen, --OR.sup.e, --NR.sup.e.sub.2, --SR.sup.e, and
--P(O)(OH).sub.2, wherein R.sup.e is selected from a group
consisting of --H, a lower alkyl, and an aryl; or each Y is
independently selected from a group consisting of CH.sub.2glycinyl,
CH.sub.2NHethoxy, CH.sub.2NHCH.sub.2alkyl, CH.sub.2NHCH.sub.2t-Bu,
CH.sub.2NHCH.sub.2aryl, and CH.sub.2NHCH.sub.2-substituted aryl; or
when n is 2, each Y is taken together to form a fused aromatic ring
system; and wherein when m=n=2, Y is not phenyl or
4-hydroxyphenyl.
3. The method of claim 2, wherein the compound III has the
structure selected from a group consisting of: ##STR00211##
##STR00212##
4. The method of claim 2, wherein the compound III has the
structure: ##STR00213##
5. The method of claim 2, wherein the compound III has the
structure: ##STR00214##
6. The Method of claim 2, wherein the compound III has the
structure: ##STR00215##
7. The method of claim 2, wherein the compound III has the
structure: ##STR00216##
8. The method of claim 2, wherein the compound III has the
structure: ##STR00217##
9. The method of claim 2, wherein the compound III has the
structure: ##STR00218##
10. The method of claim 2, wherein the compound III has the
structure: ##STR00219##
11-12. (canceled)
13. The method of claim 1, wherein the disorder or disease is
cancer or tumor.
14. The method of claim 13, wherein the compound III is
administered in combination with an effective amount of a compound
selected from a group consisting of a therapeutic antibody, a
chemotherapeutic agent, and an immunotoxic agent, or any
combination thereof.
15. The method of claim 13, wherein the cancer is selected from a
group consisting of an alimentary/gastrointestinal tract cancer,
colon cancer, liver cancer, skin cancer, breast cancer, ovarian
cancer, prostate cancer, lymphoma, leukemia, kidney cancer, lung
cancer, muscle cancer, bone cancer, bladder cancer, and brain
cancer.
16. The method of claim 15, wherein the cancer is colon cancer or
lung cancer.
17. The method of claim 14, wherein the therapeutic agent is
selected from a group consisting of an antimetabolite; a DNA
cross-linking agent; an alkylating agent; a topoisomerase I
inhibitor; a microtubule inhibitor, a vinca alkaloid, a
mitomycin-type antibiotic, and a bleomycin-type antibiotic.
18. The method of claim 14, wherein the chemotherapeutic agent is
selected from a group consisting of methotrexate,
cisplatin/carboplatin; canbusil; dactinomycin; taxol (paclitaxol),
antifolate, colchicine, demecolcine, etoposide, taxane/taxol,
docetaxel, doxorubicin, anthracycline antibiotic, daunorubicin,
caminomycin, epirubicin, idarubicin, mitoxanthrone,
4-demethoxy-daunomycin, 11-deoxydaunorubicin, 13-deoxydaunorubicin,
adriamycin-14-benzoate, adriamycin-14-octanoate, and
adriamycin-14-naphthaleneacetate.
19-21. (canceled)
22. The method of claim 1, wherein the disorder or disease is
retinopathy or a vitreoretinal disease.
23-40. (canceled)
41. A method for treating a subject having, or at risk of
developing asthma, comprising administering to a subject in need
thereof an effective amount of at least one compound having the
following structure or tautomers or pharmaceutically acceptable
salts thereof: ##STR00220##
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation application of U.S.
application Ser. No. 10/679,209 filed Oct. 2, 2003, now pending;
which claims the benefit under 35 USC .sctn.119(e) of U.S.
Application Ser. Nos. 60/479,295 filed Jun. 17, 2003, 60/466,983
filed Apr. 30, 2003, 60/463,818 filed Apr. 17, 2003, 60/443,752
filed Jan. 29, 2003, 60/440,234 filed Jan. 14, 2003 and 60/415,981
filed Oct. 3, 2002. This disclosure each of the prior applications
is considered part of and is incorporated by reference in the
disclosure of this application.
FIELD OF THE INVENTION
[0002] The present invention relates generally to treating
disorders associated with vascular functioning, and more
specifically to compounds and methods of treating such
disorders.
BACKGROUND OF THE INVENTION
[0003] The vascular system is a prime mediator of homeostasis,
playing key roles in the maintenance of normal physiologic
functioning. For example, the vascular endothelium's barrier
function serves to regulate the entry of fluid, electrolytes, and
proteins into tissues, blood vessel tone contributes to the
regulation of tissue perfusion, and the vascular endothelium's low
mitotic index contributes to the regulation of tissue growth. The
term "vasculostasis" refers to the maintenance of this homeostatic
vascular functioning, and "vasculostatic agents" as agents that
seek to address conditions in which vasculostasis is compromised by
preventing the loss of or restoring or maintaining
vasculostasis.
[0004] Compromised vasculostasis has serious pathologic
consequences. For example, if vascular permeability increases
beyond manageable levels, the resulting edema may negatively impact
tissue and organ function and ultimately survival. Examples where
excessive vascular permeability leads to particularly deleterious
effects include pulmonary edema, cerebral edema, and cardiac edema
(Ritchie AC: Boyd's Textbook of Pathology. London Lea and Febiger,
1990). In general, however, edema in any tissue or organ leads to
some loss of normal function, and therefore to the risk of
morbidity or even mortality. Similarly, excessive endothelial
proliferation may damage tissues (such as the retina in
proliferative retinopathies) or fuel unwanted tissue growth (such
as with tumor growth).
[0005] Many pathologic and disease situations are marked by
multiple disregulations in vasculostasis. Angiogenesis, for
example, encompasses both enhanced vascular proliferation and
permeability, as newly-formed blood vessels do not generally
exhibit the same level of vascular barrier function as
well-established or mature vessels. Examples of such
hyper-permeable vasculature can be found in cancers,
vasculoproliferative diseases, retinal diseases, and rheumatoid
arthritis. The connection between angiogenesis and
hyperpermeability may partly result from the dual action of factors
such as vascular endothelial growth factor (VEGF), which induces
both endothelial proliferation and vascular permeability. This
connection may also reflect the immature nature of angiogenic
vessels, in which the intracellular and/or extracellular structures
or mechanisms that establish normal vascular barrier function have
not yet fully formed. It may also be the case that angiogenesis and
vascular permeability are linked by a co-dependence on common
cellular mechanisms, for example in the case of cellular junction
disassembly which would serve to enhance both paracellular
permeability and cellular migration (both being components of the
angiogenic process). A comprehensive treatment for many diseases,
then, might involve vasculostatic agents that act upon one or more
components of vasculostasis disregulation (based, for example, upon
their level of action along intracellular signaling cascades). One
such example would be a single therapeutic agent that impacts both
angiogenesis and vascular permeability.
[0006] One way of impacting vasculostasis is by influencing
endothelial cell responses to environmental signals (such as
hypoxia) or vasoactive agents. For example, the vascular
endothelium regulates fluid balance by adjusting both transcellular
permeability (movement of fluid and proteins across endothelial
cells via a network of vesicles) and paracellular permeability
(movement of fluid and proteins between inter-endothelial cell
junctions). Edema is most commonly thought to result from a
breakdown in the inter-endothelial cell barrier, leading to
increased paracellular permeability at the capillary and
postcapillary venule level. Mechanistically, paracellular vascular
leakage results from a breakdown in inter-cellular junctional
integrity, via the dissolution of tight junctions and coupled to
changes in cytoskeletal support elements that maintain normal
cell-to-cell apposition. Several vasoactive mediators can trigger
dissolution of these cellular elements, including histamine,
bradykinin, thrombin, nitric oxide, eicosanoids (e.g., thromboxanes
and leukotrienes), platelet activating factor (PAF), tumor necrosis
factor (TNF), interleukins (e.g., IL-1 and IL-6), hepatocyte growth
factor (HGF), and vascular endothelial growth factor (VEGF). Using
VEGF as an example, the sequence of events that lead to vascular
leakage is generally believed to be as follows: reduced blood flow
(e.g., as a result of thrombus formation) leads to tissue hypoxia,
which leads to the upregulation of VEGF production, which leads to
induction of vascular leakage. This VEGF effect is at the level of
the endothelial cell, in other words VEGF binding to specific VEGF
receptors expressed on endothelial cells leads to a cascade of
intracellular events culminating in the loss of normal
intercellular barrier function. Therefore, by affecting these
intracellular events, vasulostatic agents could counter the
negative effects of environmental signals such as hypoxia or
vasoactive mediators such as VEGF, and thereby work to restore
vasculostasis.
[0007] The cascade of events that leads to the loss of endothelial
barrier function is complex and incompletely understood. Data
support a role for kinases as at least one aspect of this process.
For example, VEGF-mediated edema has been shown to involve
intracellular signaling by Src family kinases, protein kinase C,
and Akt kinase. Kinases are believed to mediate the phosphorylation
of junctional proteins such as beta-catenin and vascular
endothelial (VE)-cadherin, leading to the dissolution of adherens
junctions and the dissociation of cadherin-catenin complexes from
their cytoskeletal anchors. In addition, proteins which regulate
the intercellular contractile machinery such as myosin light chain
kinase (MLCK) and myosin light chain (MLC) are also activated,
resulting in cellular contraction, and therefore an opening of
intercellular junctions.
[0008] Maintaining or restoring vasculostasis should be beneficial
to overall patient outcome in situations such as inflammation,
allergic diseases, cancer, cerebral stroke, myocardial infarction,
pulmonary and cardiac insufficiency, renal failure, and
retinopathies, to name a few. In addition, edema formation is a
recognized but unwanted consequence of many therapeutic
interventions, such as immunotherapy, cancer chemotherapy and
radiation therapy, therefore vasculostatic agents that inhibit
vascular permeability could be used in a co-therapy approach to
reduce the deleterious side-effects of such therapies. Furthermore,
in many cases edema formation causes uneven delivery of therapeutic
agents to diseased tissues, therefore vasculostatic agents that
inhibit vascular permeability could be used in a co-therapy
approach to enhance delivery and efficacy of such therapies.
Finally, as edema is a general consequence of tissue hypoxia, it
can also be concluded that inhibition of vascular leakage
represents a potential approach to the treatment of tissue hypoxia.
For example, interruption of blood flow by pathologic conditions
(such as thrombus formation) or medical intervention (such as
cardioplegia, organ transplantation, and angioplasty) or physical
trauma, could be treated both acutely and prophylactically using
vasculostatic agents that reduce vascular permeability.
SUMMARY OF THE INVENTION
[0009] The present invention is based on the discovery that certain
chemical compounds are effective vasculostatic agents. Compounds of
the invention are effective for the treatment of such indications
as myocardial infarction (MI), stroke, ischemia or reperfusion
related tissue injury and cancer, for example. Thus, compositions
and methods are provided for treating disorders associated with
compromised vasculostasis, examples of which are edema resulting
from excess vascular permeability or vascular leakage and
angiogenesis associated with retinal diseases and cancer. Some of
the compounds described herein are effective kinase inhibitors,
including but not limited to tyrosine, serine or threonine kinase
inhibitors, for example, Src-family inhibitors.
[0010] Such vasculostatic agents, alone or in combination with
other agents, are effective in blocking vascular permeability or
leakage or angiogenesis. In one embodiment, the invention provides
a composition containing a therapeutically effective amount of a
compound of the invention in a pharmaceutically acceptable
carrier.
[0011] In one embodiment, the invention provides a method for
treating a disorder associated with compromised vasculostasis in a
subject, comprising administering to a subject in need thereof an
effective amount of a compound that is a vasculostatic agent. In an
illustrative example, the method includes use of at least one of
the compounds as set forth in Structures I, II, III, IIIa, IV, V,
VI or VII or any combination thereof. In one aspect, the compound
is set forth in FIG. 1.
[0012] In one embodiment, compounds are provided having the
structure (I):
##STR00001## [0013] wherein: [0014] each R.sub.0 is independently
--H, --COOH, --OR', --SO.sub.3H, wherein R' is --H or lower alkyl,
or when x=2, each R.sub.o is taken together to form a 1,3-dioxolyl
ring, or [0015] each R.sub.0 is independently alkyl, substituted
alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,
cycloalkyl, substituted cycloalkyl, heterocyclic, substituted
heterocyclic, aryl, substituted aryl, heteroaryl, substituted
heteroaryl, alkylaryl, substituted alkylaryl, arylalkyl,
substituted arylalkyl, arylalkenyl, substituted arylalkenyl,
arylalkynyl, substituted arylalkynyl, halogen, amino, amido, nitro,
or thioalkyl, [0016] R.sub.1 and R.sub.2 are each independently
hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl,
alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,
heterocyclic, substituted heterocyclic, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, alkylaryl, substituted
alkylaryl, arylalkyl, substituted arylalkyl, arylalkenyl,
substituted arylalkenyl, arylalkynyl, or substituted arylalkynyl,
[0017] G is NH, O, S, or (CR''.sub.2).sub.p, wherein R'' is --H,
lower alkyl, or acetamido, and wherein p is 0-3, [0018] Ar is aryl
or heteroaryl, and [0019] x and y are each independently 1-4.
[0020] In another embodiment, compounds are provided having the
structure (II):
##STR00002## [0021] wherein R.sub.0, R.sub.1, R.sub.2, x, and y are
as defined above.
[0022] In yet another embodiment, compounds are provided having the
structure (III):
##STR00003## [0023] wherein: [0024] Z.sub.1-Z.sub.6 are each
independently C, --C.dbd.O, N, or NR.sup.a, wherein R.sup.a is --H,
alkyl, or substituted alkyl, wherein said substituents are halogen,
hydroxy, oxo, or amino, [0025] each X is independently halogen,
--OR.sup.b, --NR.sup.b.sub.2, or --SR.sup.b, wherein R.sup.b is --H
lower alkyl, --(CH.sub.2).sub.2NH(CH.sub.2CH.sub.3),
--(CH.sub.2).sub.3morpholyn-1-yl,
--(CH.sub.2).sub.3(N-methylpiperazinyn-1-yl), aryl, heteroaryl,
--(NH--NH--R.sup.c), --(N.dbd.N--NH--R.sup.c), wherein R.sup.c is H
or lower alkyl, [0026] each Y is independently --OR.sup.d,
--NR.sup.d.sub.2, --SR.sup.d, or --OPO.sub.3H.sub.2 wherein R.sup.d
is H, lower alkyl, aryl, heteroaryl,
--(CH.sub.2).sub.2NH(CH.sub.2CH.sub.3),
--(CH.sub.2).sub.3morpholyn-1-yl, or
--(CH.sub.2).sub.3(N-methylpiperazinyn-1-yl); or [0027] each Y is
independently alkyl, substituted alkyl, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, or halogen, wherein said
substituents are selected from halogen, --OR.sup.e,
--NR.sup.e.sub.2, --SR.sup.e, --P(O)(OH).sub.2, wherein R.sup.e is
--H, lower alkyl, aryl, or heteroaryl; or each Y is independently
CH.sub.2glycinyl, CH.sub.2NHethoxy, CH.sub.2NHCH.sub.2alkyl,
CH.sub.2NHCH.sub.2t-Bu, CH.sub.2NHCH.sub.2aryl,
CH.sub.2NHCH.sub.2substituted aryl, CH.sub.2NHCH.sub.2heteroaryl,
CH.sub.2NHCH.sub.2substituted heteroaryl; or when n is 2, each Y is
taken together to form a fused aromatic or heteroaromatic ring
system; and [0028] m and n are each independently 1 to 4, [0029]
wherein when Z.sub.1, Z.sub.3, Z.sub.5, and Z.sub.6 are each N, X
is NH.sub.2, and m=n=2, Y is not phenyl or 4-hydroxyphenyl, or
tautomers thereof.
[0030] In still another embodiment, compounds are provided having
the structure (IV):
##STR00004## [0031] wherein: [0032] L is an arylene, substituted
arylene, oxyarylene, thioalkylene, substituted thioalkylene, or
substituted oxyarylene linking moiety, [0033] C is 5- or 6-membered
aromatic or heteroaromatic ring, [0034] each X is independently H,
OR, NR.sub.2, or SR, wherein R is H or lower alkyl, [0035]
Z.sub.1-Z.sub.4 are each independently CH or N, and [0036] m is 1
to 4.
[0037] In still another embodiment, compounds are provided having
the structure (V):
##STR00005## [0038] wherein: [0039] R.sub.1, x, and y are as
defined above, [0040] R.sub.3 is --H, --SO.sub.3H, or
--SO.sub.2NMe.sub.2, [0041] M is NH, CO, SO.sub.2, (CH.sub.2)p,
wherein p is 0 to 2, [0042] G is aryl or heteroaryl, and [0043] x
and y are each independently 0-4.
[0044] In a further embodiment, there are provided methods for
treating disorders associated with compromised vasculostasis,
including administering to a subject in need thereof an effective
amount of a compound having the structure (VI):
##STR00006## [0045] wherein: [0046] A and B are each independently
5- or 6-membered aromatic rings, wherein at least one of A and B is
an aromatic heterocyclic ring having at least one heteroatom in the
heterocyclic ring, [0047] each X is independently --H, OR,
NR.sub.2, or SR, wherein R is H or lower alkyl, [0048] each Y is
independently hydrogen, alkyl, substituted alkyl, alkenyl, [0049]
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
[0050] substituted cycloalkyl, heterocyclic, substituted
heterocyclic, aryl, [0051] substituted aryl, heteroaryl,
substituted heteroaryl, alkylaryl, substituted alkylaryl,
arylalkyl, substituted arylalkyl, arylalkenyl, substituted
arylalkenyl, arylalkynyl, [0052] substituted arylalkynyl, or oxo,
with the proviso that at least one Y is not hydrogen, or [0053]
when n is 2, each Y is taken together to form a fused aromatic ring
system comprising at least one aromatic ring, and [0054] m and n
are each independently 1 to 4, thereby treating the disorder.
[0055] In yet another embodiment, invention methods include
administering to a subject in need thereof an effective amount of a
compound having the structure (VII):
##STR00007## [0056] wherein: [0057] A, B, C, and D are each
independently C, N, O, or S, [0058] each X is independently OR,
NR.sub.2, or SR, wherein R is H or lower alkyl, [0059] each Y is
independently hydrogen, alkyl, substituted alkyl, [0060] alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
[0061] substituted cycloalkyl, heterocyclic, substituted
heterocyclic, aryl, [0062] substituted aryl, heteroaryl,
substituted heteroaryl, alkylaryl, [0063] substituted alkylaryl,
arylalkyl, substituted arylalkyl, arylalkenyl, [0064] substituted
arylalkenyl, arylalkynyl, substituted arylalkynyl, with the proviso
that at least one Y is not hydrogen, and [0065] m and n are each
independently 1 to 4, [0066] thereby treating the disorder.
[0067] In another embodiment, the invention provides a method for
treating a disorder associated with compromised vasculostasis,
comprising administering to a subject in need thereof an effective
amount of a compound having the structure:
##STR00008## [0068] wherein: [0069] each X is independently H, OR,
NR.sub.2, or SR, wherein R is H or lower alkyl, [0070] each Y is
independently hydrogen, alkyl, substituted alkyl, alkenyl [0071]
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
[0072] substituted cycloalkyl, heterocyclic, substituted
heterocyclic, aryl, [0073] substituted aryl, heteroaryl,
substituted heteroaryl, alkylaryl, [0074] substituted alkylaryl,
arylalkyl, substituted arylalkyl, arylalkenyl, [0075] substituted
arylalkenyl, arylalkynyl, substituted arylalkynyl, aroyl,
substituted [0076] aroyl, acyl, or substituted acyl, with the
proviso that at least one Y is not hydrogen, or [0077] when n is 2,
each Y is taken together to form a fused aromatic ring system
comprising at least one aromatic ring, [0078] m is 1 to 4, and
[0079] n is 1 or 2, thereby treating the disorder.
[0080] In another embodiment, the invention provides a method for
treating a disorder associated with compromised vasculostasis,
comprising administering to a subject in need thereof an effective
amount of a compound having the structure:
##STR00009## [0081] wherein: [0082] each X is independently H, OR,
NR.sub.2, or SR, wherein R is H or lower alkyl, [0083] each Y is
independently hydrogen, alkyl, substituted alkyl, alkenyl
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, heterocyclic, substituted heterocyclic,
aryl, substituted aryl, heteroaryl, substituted heteroaryl,
alkylaryl, substituted alkylaryl, arylalkyl, substituted arylalkyl,
arylalkenyl, substituted arylalkenyl, arylalkynyl, substituted
arylalkynyl, aroyl, substituted aroyl, acyl, or substituted acyl,
with the proviso that at least one Y is not hydrogen, or [0084]
when n is 2, each Y is taken together to form a fused aromatic ring
system comprising at least one aromatic ring, and [0085] m and n
are each independently 1 or 2.
[0086] In another embodiment, the invention provides a method for
treating a disorder associated with compromised vasculostasis,
comprising administering to a subject in need thereof an effective
amount of a compound having the structure:
##STR00010## [0087] wherein: [0088] Z is N, O, or S; [0089] each X
is independently H, OR, NR.sub.2, or SR, wherein R is H or lower
alkyl, [0090] each Y is independently hydrogen, alkyl, substituted
alkyl, alkenyl substituted alkenyl, alkynyl, substituted alkynyl,
cycloalkyl, substituted cycloalkyl, heterocyclic, substituted
heterocyclic, aryl, substituted aryl, heteroaryl, substituted
heteroaryl, alkylaryl, substituted alkylaryl, arylalkyl,
substituted arylalkyl, arylalkenyl, substituted arylalkenyl,
arylalkynyl, substituted arylalkynyl, aroyl, substituted aroyl,
acyl, or substituted acyl, with the proviso that at least one Y is
not hydrogen, or [0091] when n is 2, each Y is taken together to
form a fused aromatic ring system comprising at least one aromatic
ring, and [0092] m is 1 to 4, and [0093] n is 1 or 2.
[0094] In another embodiment, the invention provides a method for
treating a disorder associated with compromised vasculostasis
comprising administering to a subject in need thereof an effective
amount of a compound having structure (VII):
##STR00011## [0095] wherein: [0096] A, B, C, and D are each
independently C, N, O, or S, [0097] each X is independently H, OR,
NR.sub.2, or SR, wherein R is H or lower alkyl, [0098] each Y is
independently hydrogen, alkyl, substituted alkyl, [0099] alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, heterocyclic, substituted heterocyclic,
aryl, substituted aryl, heteroaryl, substituted heteroaryl,
alkylaryl, substituted alkylaryl, arylalkyl, substituted arylalkyl,
arylalkenyl, substituted arylalkenyl, arylalkynyl, substituted
arylalkynyl, aroyl, substituted aroyl, acyl, or substituted acyl,
with the proviso that at least one Y is not hydrogen, and [0100] m
and n are each independently 1 to 4, thereby treating the
disorder.
[0101] In one embodiment, the invention provides a method for
treating a disorder associated with compromised vasculostasis,
comprising administering to a subject in need thereof an effective
amount of a compound, wherein the compound is set forth in
Structures I, II, III, IIIa, IV, V, or any combination thereof. The
disorder is for example, but not limited to, myocardial infarction,
stroke, congestive heart failure, an ischemia or reperfusion
injury, cancer, arthritis or other arthropathy, retinopathy or
vitreoretinal disease, macular degeneration, autoimmune disease,
vascular leakage syndrome, inflammatory disease, edema, transplant
rejection, burn, or acute or adult respiratory distress syndrome
(ARDS).
[0102] In still another embodiment, there are provided articles of
manufacture including packaging material and a pharmaceutical
composition contained within the packaging material, wherein the
pharmaceutical composition is capable of treating a disorder
associated with compromised vasculostasis, wherein the
pharmaceutical composition comprises at least one compound having
any one of the structures as set forth above.
[0103] In one embodiment, the invention provides a pharmaceutical
composition comprising a compound as set forth in Structures I, II,
III, IIIa, IV, V, or VII, or any combination thereof, in a
pharmaceutically acceptable carrier.
[0104] In one embodiment, the invention provides an article of
manufacture comprising packaging material and a pharmaceutical
composition contained within said packaging material, wherein said
packaging material comprises a label which indicates that said
pharmaceutical composition can be used for treatment of disorders
associated with compromised vasculostasis and wherein said
pharmaceutical composition comprises a compound set forth in
Structures I, II, III, IIIa, IV, V, VI or VII, or any combination
thereof.
[0105] In one embodiment, the invention provides an article of
manufacture comprising packaging material and a pharmaceutical
composition contained within said packaging material, wherein said
packaging material comprises a label which indicates that said
pharmaceutical composition can be used for treatment of disorders
associated with vascular permeability leakage or compromised
vasculostasis selected from is myocardial infarction, stroke,
congestive heart failure, an ischemia or reperfusion injury,
cancer, arthritis or other arthropathy, retinopathy or
vitreoretinal disease, macular degeneration, autoimmune disease,
vascular leakage syndrome, inflammatory disease, edema, transplant
rejection, burns, or acute or adult respiratory distress syndrome
(ARDS) and wherein said pharmaceutical composition comprises a
compound set forth in Structures I, II, III, IIIa, IV, V, VI or
VII, or any combination thereof.
[0106] In one embodiment, the invention provides a method of
treating a compromised vasculostasis disorder, comprising the
administration of a therapeutically effective amount of at least
one compound set forth in Structures I, II, III, IIIa, IV, V, VI or
VII, or any combination thereof, or pharmaceutically acceptable
salts, hydrates, solvates, crystal forms and individual
diastereomers thereof, to a subject in need of such treatment.
[0107] In one embodiment, the invention provides a method of
treating a disorder associated with vasculostasis, comprising the
administration of a therapeutically effective amount of at least
one compound as set forth in Structures I, II, III, IIIa, IV, V, VI
or VII, or any combination thereof, or pharmaceutically acceptable
salts, hydrates, solvates, crystal forms and individual
diastereomers thereof, in combination with an anti-inflammatory,
chemotherapeutic agent, immunomodulatory agent, therapeutic
antibody or a protein kinase inhibitor, to a subject in need of
such treatment.
[0108] In one embodiment, the invention provides a method of
treating a subject having or at risk of having myocardial
infarction comprising administering to the subject a
therapeutically effective amount of a compound as set forth in
Structures I, II, III, IIIa, IV, V, VI or VII, or any combination
thereof thereby treating the subject.
[0109] In one embodiment, the invention provides a method of
treating a subject having or at risk of having vascular leakage
syndrome (VLS) comprising administering to the subject a
therapeutically effective amount of a compound as set forth in
Structures I, II, III, IIIa, IV, V, VI or VII, or any combination
thereof thereby treating the subject.
[0110] In one embodiment, the invention provides a method of
treating a subject having or at risk of having cancer comprising
administering to the subject a therapeutically effective amount of
a compound as set forth in Structures I, II, III, IIIa, IV, V, or
VII, or any combination thereof thereby treating the subject.
[0111] In one embodiment, the invention provides a method of
treating a subject having or at risk of having stroke comprising
administering to the subject a therapeutically effective amount of
a compound as set forth in Structures I, II, III, IIIa, IV, V, VI
or VII, or any combination thereof thereby treating the
subject.
[0112] In one embodiment, the invention provides a method of
treating a subject having or at risk of having ARDS comprising
administering to the subject a therapeutically effective amount of
a compound as set forth in Structures I, II, III, IIIa, IV, V, VI
or VII, or any combination thereof thereby treating the
subject.
[0113] In one embodiment, the invention provides a method of
treating a subject having or at risk of having burns comprising
administering to the subject a therapeutically effective amount of
a compound as set forth in Structures I, II, III, IIIa, IV, V, VI
or VII, or any combination thereof thereby treating the
subject.
[0114] In one embodiment, the invention provides a method of
treating a subject having or at risk of having arthritis comprising
administering to the subject a therapeutically effective amount of
a compound as set forth in Structures I, II, III, IIIa, IV, V, VI
or VII, or any combination thereof thereby treating the
subject.
[0115] In one embodiment, the invention provides a method of
treating a subject having or at risk of having edema comprising
administering to the subject a therapeutically effective amount of
a compound as set forth in Structures I, II, III, IIIa, IV, V, VI
or VII, or any combination thereof thereby treating the
subject.
[0116] In one embodiment, the invention provides a method of
treating a subject having or at risk of having vascular leakage
syndrome (VLS) comprising administering to the subject a
therapeutically effective amount of a compound as set forth in
Structures I, II, III, IIIa, IV, V, VI or VII, or any combination
thereof thereby treating the subject.
[0117] In one embodiment, the invention provides a method of
treating a subject having or at risk of having retinopathy or
vitreoretinal disease comprising administering to the subject a
therapeutically effective amount of a compound as set forth in
Structures I, II, III, IIIa, IV, V, VI or VII, or any combination
thereof thereby treating the subject.
[0118] In one embodiment, the invention provides a method of
treating a subject having or at risk of having ischemic or
reperfusion related tissue injury or damage, comprising
administering to the subject a therapeutically effective amount of
a compound as set forth in Structures I, II, III, IIIa, IV, V, VI
or VII, or any combination thereof thereby treating the
subject.
[0119] In one embodiment, the invention provides a method of
treating a subject having or at risk of having autoimmune disease,
comprising administering to the subject a therapeutically effective
amount of a compound as set forth in Structures I, II, III, IIIa,
IV, V, VI or VII, or any combination thereof thereby treating the
subject.
[0120] In one embodiment, the invention provides a method of
treating a subject having or at risk of having transplant
rejection, comprising administering to the subject a
therapeutically effective amount of a compound as set forth in
Structures I, II, III, IIIa, IV, V, VI or VII, or any combination
thereof thereby treating the subject.
[0121] In one embodiment, the invention provides a method of
treating a subject having or at risk of having inflammatory
disease, comprising administering to the subject a therapeutically
effective amount of a compound as set forth in Structures I, II,
III, IIIa, IV, V, VI or VII, or any combination thereof thereby
treating the subject.
[0122] In one embodiment, the invention provides a process for
making a pharmaceutical composition comprising combining a
combination of a compound set forth in Structures I, II, III, IIIa,
IV, V, VI or VII, or any combination thereof or its
pharmaceutically acceptable salts, hydrates, solvates, crystal
forms salts and individual diastereomers thereof and a
pharmaceutically acceptable carrier.
[0123] In one embodiment, the invention provides a pharmaceutical
composition comprising a compound as set forth in Structure I, II,
III, IIIa, IV, V, VII, or VIII in a pharmaceutically acceptable
carrier.
[0124] In one embodiment, the invention provides a method for
inhibiting or reducing vascular leakage in a subject, comprising
administering to a subject in need thereof an effective amount of
IL-2 in combination with a compound of Structure set forth in
Structures I, II, III, IIIa, IV, V, VI or VII or any combination
thereof, thereby reducing vascular leakage in the subject. In one
aspect, the compound may be N-(2-(1H-Indol-2-yl)-phenyl)-phthalamic
acid or 6,7-bis-(3-hydroxyphenyl)-pteridine-2,4-diamine.
[0125] In one embodiment, the invention provides a pharmaceutical
composition comprising IL-2 and at least one compound as set forth
in Structures I, II, III, IIIa, IV, V, VI or VII or any combination
thereof, in a concentration effective to reduce vascular leakage
associated with IL-2 administration.
[0126] In one embodiment, the invention provides a method for
treating cancer or a tumor in a subject, comprising administering
to a subject in need thereof an effective amount of a therapeutic
antibody, chemotherapeutic agent or immunotoxic agents, in
combination with a compound set forth in Structures I, II, III,
IIIa, IV, V, VI or VII or any combination thereof, thereby treating
the cancer or tumor in the subject.
[0127] In one embodiment, the invention provides a pharmaceutical
composition comprising a therapeutic agent and at least one
compound as set forth in Structures I, II, III, IIIa, IV, V, VI or
VII or any combination thereof, in a concentration effective to
treat cancer in a subject. The cancer may be any cancer, including
but not limited to an alimentary/gastrointestinal tract cancer,
colon cancer, liver cancer, skin cancer, breast cancer, ovarian
cancer, prostate cancer, lymphoma, leukemia, kidney cancer, lung
cancer, muscle cancer, bone cancer, bladder cancer or brain
cancer.
[0128] In one embodiment, the invention provides a method for
treating a T-cell mediated disorder, comprising the administration
of a therapeutically effective amount of at least one compound set
forth in Structures I, II, III, IIIa, IV, V, VI or VII, or any
combination thereof or pharmaceutically acceptable salts, hydrates,
solvates, crystal forms salts and individual diastereomers thereof,
to a subject in need of such treatment.
BRIEF DESCRIPTION OF THE FIGURES
[0129] FIGS. 1A-1F shows exemplary compounds of the invention.
[0130] FIG. 2 shows the results of
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine, sulfate salt and
doxorubicin for treatment of lung metastases. Syngeneic Lewis lung
carcinoma cells were injected I.V. in order to establish lung
metastases in Balb/C mice. Beginning 10 days after cells were
injected, doxorubicin (3 mg/kg) and/or
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine, sulfate salt (various
doses as shown) was given I.P. every 3 days for 3 cycles. Animals
were sacrificed at day 20, lungs were collected, and weighed. Net
tumor burden is the weight of tumor-bearing lungs minus the average
weight of normal control lungs. N=5/group, p<0.02.
[0131] FIG. 3 illustrates the effect of compounds administered in
conjunction with doxorubicin in an in vivo model of metastatic
colon cancer (CT-26 adenocarcinoma). Syngeneic CT-26 Colon
carcinoma cells were injected I.V. in order to establish lung
metastases in Balb/C mice. Beginning 10 days after cells were
injected, indicated test agents were given I.P. every 3 days for 3
cycles. Animals were sacrificed at day 20, lungs were collected,
and weighed. Net tumor burden is the weight of tumor-bearing lungs
minus the average weight of normal control lungs. N=5/group,
p<0.02. In these graphs, compound A is
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine sulfate salt, and
compound B is 6,7-diphenyl-pteridine-2,4-diamine.
[0132] FIG. 4 illustrates the effects of compounds of the present
invention for co-drug therapy with Taxotere as described herein.
Syngeneic CT-26 Colon carcinoma cells were used in order to
establish lung metastases in Balb/C mice as described for FIG. 3.
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine, sulfate salt (compound
A) and 6,7-diphenyl-pteridine-2,4-diamine (compound B) from FIG. 1
are shown in FIG. 4.
[0133] FIG. 5 shows a photo of representative lung samples from the
experiment shown in FIG. 4 with 6,7-diphenyl-pteridine-2,4-diamine
(compound B) and doxorubicin.
[0134] FIG. 6 illustrates the effect of compounds administered in
conjunction with docetaxel in the in vivo model of metastatic colon
cancer (CT-26 adenocarcinoma) described for FIG. 4.
2,3-Bis(3,4-dihydroxyphenyl)-pyrido[2,3-b]pyrazin-6-ylamine
dihydrochloride salt (compound C) from FIG. 1 is shown in FIG. 6 as
compound C. N=5/group, p<0.02.
[0135] FIGS. 7 and 8 illustrate the effects of compounds of the
invention for their capacity to inhibit IL-2 induced VLS. The
graphs present representative examples of compounds cited in this
application and their effects on VLS. In the graphs, compound D is
N-(2-(1H-indol-2-yl)-phenyl)-phthalamic acid and compound E is
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine.
[0136] FIG. 9 illustrates the effects of compounds of the invention
for their effect on IL-2 induced anti-tumor actions. The graph
presents representative examples of compounds cited in this
application and their effects on IL-2 mediated reductions in
metastatic melanoma tumor burden. In the graphs, compound D is
N-(2-(1H-indol-2-yl)-phenyl)-phthalamic acid and compound E is
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine. Invention compound
concentrations are listed in parenthesis in mg/kg while IL-2
concentration is given in parenthesis kilounits.
[0137] FIGS. 10 and 11 illustrate the effects of compounds of the
invention for their capacity to inhibit IL-2 induced T-cell
proliferation. The graphs present representative examples of
compounds cited in this application and their effects on T-cell
proliferation. In the graphs, compound D is
N-(2-(1H-indol-2-yl)-phenyl)-phthalamic acid and compound E is
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine.
[0138] FIG. 12 illustrates the effects of invention compounds for
their capacity to inhibit edema associated with Acute Respiratory
Distress Syndrome (ARDS). NIH Swiss mice were given an
intraperitoneal injection of 1.5 mg/kg Oleic Acid of (in this
example formulated in saline) and/or invention compounds. Four
hours subsequent to injection animals were sacrificed followed by
collection, blotting and weighing (wet weight) of the lungs. Lungs
were then dried at 80.degree. C. for 24 hours and weighed (dry
weight). N=4/group, 6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine,
sulfate salt (compound E--in the 0.5 mg/kg range, in this example
formulated in 50% PEG400:50% water) typically reduced
ARDS-associated edema by >50% while
4-[4-amino-6-(3,4-dihydroxyphenyl)pteridin-7-yl]benzene-1,2-diol
(compound F--in the 0.5 mg/kg range, in this example formulated in
50% PEG400:50% water) typically reduced ARDS-induced edema by
>100%.
[0139] FIGS. 13 and 14 illustrate the effects of invention
compounds for their capacity to inhibit angiogenesis in vivo. The
graph presents representative examples of compounds cited in this
application which successfully inhibited angiogenesis in vivo.
Tumor extracellular matrix infused with the 160 ng of the described
growth factors were injected subcutaneously in a Balb/C mouse. The
described invention compound was injected daily at the described
concentration for 5 days. After 5 days the animals were sacrificed
and angiogenesis quantified based on the binding of fluorescently
labeled, endothelium specific FITC-lectin. In the graph, compound A
is 6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine sulfate salt.
DETAILED DESCRIPTION OF THE INVENTION
[0140] The invention provides compounds which are vasculostatic
agents and methods of use thereof. Invention compounds are useful
in treating a variety of disorders, including but not limited to
myocardial infarction, stroke, cancer, vascular leakage syndrome
(VLS), ocular and retinal disease, bone disease, pleural effusion,
edema, and ischemia. The term "vasculostasis" is hereby defined as
referring to the maintenance of a homeostatic vascular functioning,
and "vasculostatic agents" as agents that seek to address
conditions in which vasculostasis is compromised by preventing the
loss of or restoring or maintaining vasculostasis.
[0141] In one embodiment, the present invention provides compounds
of structure (I):
##STR00012## [0142] wherein: [0143] each R.sub.0 is independently
--H, --COOH, --OR', --SO.sub.3H, wherein R' is --H or lower alkyl,
or when x=2, each R.sub.o is taken together to form a 1,3-dioxolyl
ring, or [0144] each R.sub.0 is independently alkyl, substituted
alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,
cycloalkyl, substituted cycloalkyl, heterocyclic, substituted
heterocyclic, aryl, substituted aryl, heteroaryl, substituted
heteroaryl, alkylaryl, substituted alkylaryl, arylalkyl,
substituted arylalkyl, arylalkenyl, substituted arylalkenyl,
arylalkynyl, substituted arylalkynyl, halogen, amino, amido, nitro,
or thioalkyl, [0145] R.sub.1 and R.sub.2 are each independently
hydrogen, alkyl, substituted alkyl, alkenyl substituted alkenyl,
alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,
heterocyclic, substituted heterocyclic, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, alkylaryl, substituted
alkylaryl, arylalkyl, substituted arylalkyl, arylalkenyl,
substituted arylalkenyl, arylalkynyl, or substituted arylalkynyl,
[0146] G is NH, O, S, or (CR''.sub.2).sub.p, wherein R'' is --H,
lower alkyl, or acetamido, and wherein p is 0-3, [0147] Ar is aryl
or heteroaryl, and [0148] x and y are each independently 0-4.
[0149] In one embodiment, R.sub.0 is --COOH, x=1, and each R.sub.1
and R.sub.2 is hydrogen.
[0150] Exemplary compounds of structure I include:
##STR00013## ##STR00014## ##STR00015## ##STR00016##
[0151] In another embodiment of the invention, there are provided
compounds of structure (II):
##STR00017## [0152] wherein: [0153] wherein R.sub.0, R.sub.1, and
R.sub.2, x, and y are as defined above.
[0154] In one embodiment, R.sub.0 is --COOH, x=1, and R.sub.1 and
R.sub.2 are each hydrogen.
[0155] In yet another embodiment of the invention, there are
provided compounds of structure (III):
##STR00018## [0156] wherein: [0157] Z.sub.1-Z.sub.6 are each
independently C, --CO.dbd.N, or NR.sup.a, wherein R.sup.a is --H,
alkyl, or substituted alkyl, wherein said substituents are halogen,
hydroxy, oxo, or amino, [0158] each X is independently halogen,
--OR.sup.b, --NR.sup.b.sub.2, or --SR.sup.b, wherein R.sup.b is --H
lower alkyl, --(CH.sub.2).sub.2NH(CH.sub.2CH.sub.3),
--(CH.sub.2).sub.3morpholyn-1-yl,
--(CH.sub.2).sub.3(N-methylpiperazinyn-1-yl), aryl, heteroaryl,
--(NH--NH--R.sup.c), --(N.dbd.N--NH--R.sup.c), wherein R.sup.c is H
or lower alkyl, [0159] each Y is independently --OR.sup.d,
--NR.sup.d.sub.2, --SR.sup.d, or --OPO.sub.3H.sub.2 wherein R.sup.d
is H, lower alkyl, aryl, heteroaryl,
--(CH.sub.2).sub.2NH(CH.sub.2CH.sub.3),
--(CH.sub.2).sub.3morpholyn-1-yl, or
--(CH.sub.2).sub.3(N-methylpiperazinyn-1-yl); or [0160] each Y is
independently alkyl, substituted alkyl, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, or halogen, wherein said
substituents are selected from halogen, --OR.sup.e,
--NR.sup.e.sub.2, --SR.sup.e, --P(O)(OH).sub.2, wherein R.sup.e is
--H, lower alkyl, aryl, or heteroaryl; or each Y is independently
CH.sub.2glycinyl, CH.sub.2NHethoxy, CH.sub.2NHCH.sub.2alkyl,
CH.sub.2NHCH.sub.2t-Bu, CH.sub.2NHCH.sub.2aryl,
CH.sub.2NHCH.sub.2substituted aryl, CH.sub.2NHCH.sub.2heteroaryl,
CH.sub.2NHCH.sub.2substituted heteroaryl; or when n is 2, each Y is
taken together to form a fused aromatic or heteroaromatic ring
system; and [0161] m and n are each independently 1 to 4,
[0162] wherein when Z.sub.1, Z.sub.3, Z.sub.5, and Z.sub.6 are each
N, X is NH.sub.2, and m=n=2, Y is not phenyl or
4-hydroxyphenyl,
or tautomers thereof.
[0163] Exemplary compounds of structure III include pteridines and
quinoxalines, such as
##STR00019##
[0164] Particularly effective vasculostatic agents of structure
(III) include compounds bearing hydroxy-substituted aryl rings.
Exemplary compounds according to this embodiment are set forth
below:
##STR00020##
[0165] An additional exemplary compound of structure (III) is set
forth below:
##STR00021##
[0166] Additional exemplary compounds of structure (III) include
pteridines having the structure:
##STR00022##
wherein when X.sub.1.dbd.X.sub.2=--NHR, wherein R is --H, aryl, or
substituted aryl, Y.sub.1 and Y.sub.2 include but are not limited
to the following structures III-1 to III-24:
TABLE-US-00001 Structure Y.sub.1 Y.sub.2 III-1 C.sub.6H.sub.5 H
III-2 H C.sub.6H.sub.5 III-3 C.sub.6H.sub.5 C.sub.6H.sub.5 III-4
4-C.sub.6H.sub.4OH H III-5 H 4-C.sub.6H.sub.4OH III-6
3,4-C.sub.6H.sub.3(OH).sub.2 H III-7 H 3,4-C.sub.6H.sub.3(OH).sub.2
III-8 4-C.sub.6H.sub.4F C.sub.6H.sub.5 III-9 C.sub.6H.sub.5
4-C.sub.6H.sub.4F III-10 4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-11
C.sub.6H.sub.5 4-C.sub.6H.sub.4Br III-12 4-C.sub.6H.sub.4OPh
C.sub.6H.sub.5 III-13 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-14
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-15 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-16 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-17 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-18
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-19 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-20 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-21 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-22
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-23
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-24
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group
[0167] Further exemplary pteridines have the structure
X.sub.1.dbd.X.sub.2.dbd.OR, wherein R is --H, aryl, or substituted
aryl, and Y.sub.1 and Y.sub.2 include but are not limited to the
following the structures III-25 to III-48:
TABLE-US-00002 Structure Y.sub.1 Y.sub.2 III-25 C.sub.6H.sub.5 H
III-26 H C.sub.6H.sub.5 III-27 C.sub.6H.sub.5 C.sub.6H.sub.5 III-28
4-C.sub.6H.sub.4OH H III-29 H 4-C.sub.6H.sub.4OH III-30
3,4-C.sub.6H.sub.3(OH).sub.2 H III-31 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-32 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-33 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-34
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-35 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-36 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5 III-37
C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-38 4-C.sub.6H.sub.4OH
C.sub.6H.sub.5 III-39 C.sub.6H.sub.5 4-C.sub.6H.sub.4OH III-40
C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N (pyr) III-41
4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-42 3-C.sub.6H.sub.4F
3-C.sub.6H.sub.4F III-43 4-C.sub.6H.sub.4OMe 4-C.sub.6H.sub.4OMe
III-44 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe III-45
4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-46 3-C.sub.6H.sub.4OH
3-C.sub.6H.sub.4OH III-47 3,4-C.sub.6H.sub.3(OH).sub.2
3,4-C.sub.6H.sub.3(OH).sub.2 III-48 Y.sub.1 and Y.sub.2 taken
together to form a phenathrolinyl group
[0168] Further exemplary pteridines have the structure
X.sub.1.dbd.OR and X.sub.2.dbd.NHR, wherein R is --H, aryl or
substituted aryl, and Y.sub.1 and Y.sub.2 include but are not
limited to the following structures
TABLE-US-00003 Structure Y.sub.1 Y.sub.2 III-49 C.sub.6H.sub.5 H
III-50 H C.sub.6H.sub.5 III-51 C.sub.6H.sub.5 C.sub.6H.sub.5 III-52
4-C.sub.6H.sub.4OH H III-53 H 4-C.sub.6H.sub.4OH III-54
3,4-C.sub.6H.sub.3(OH).sub.2 H III-55 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-56 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-57 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-58
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-59 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-60 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5 III-61
C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-62 4-C.sub.6H.sub.4OH
C.sub.6H.sub.5 III-63 C.sub.6H.sub.5 4-C.sub.6H.sub.4OH III-64
C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N (pyr) III-65
4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-66 3-C.sub.6H.sub.4F
3-C.sub.6H.sub.4F III-67 4-C.sub.6H.sub.4OMe 4-C.sub.6H.sub.4OMe
III-68 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe III-69
4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-70 3-C.sub.6H.sub.4OH
3-C.sub.6H.sub.4OH III-71 3,4-C.sub.6H.sub.3(OH).sub.2
3,4-C.sub.6H.sub.3(OH).sub.2 III-72 Y.sub.1 and Y.sub.2 taken
together to form a phenathrolinyl group
[0169] Further exemplary pteridines have the structure
X.sub.1.dbd.NHR and X.sub.2.dbd.OR, wherein R is --H, aryl or
substituted aryl, and Y.sub.1 and Y.sub.2 include but are not
limited to the following structures
TABLE-US-00004 Structure Y.sub.1 Y.sub.2 III-73 C.sub.6H.sub.5 H
III-74 H C.sub.6H.sub.5 III-75 C.sub.6H.sub.5 C.sub.6H.sub.5 III-76
4-C.sub.6H.sub.4OH H III-77 H 4-C.sub.6H.sub.4OH III-78
3,4-C.sub.6H.sub.3(OH).sub.2 H III-79 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-80 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-81 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-82
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-83 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-84 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5 III-85
C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-86 4-C.sub.6H.sub.4OH
C.sub.6H.sub.5 III-87 C.sub.6H.sub.5 4-C.sub.6H.sub.4OH III-88
C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N (pyr) III-89
4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-90 3-C.sub.6H.sub.4F
3-C.sub.6H.sub.4F III-91 4-C.sub.6H.sub.4OMe 4-C.sub.6H.sub.4OMe
III-92 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe III-93
4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-94 3-C.sub.6H.sub.4OH
3-C.sub.6H.sub.4OH III-95 3,4-C.sub.6H.sub.3(OH).sub.2
3,4-C.sub.6H.sub.3(OH).sub.2 III-96 Y.sub.1 and Y.sub.2 taken
together to form a phenathrolinyl group
[0170] Additional exemplary pteridines have the structure
##STR00023##
[0171] wherein X.sub.1.dbd.NHR, wherein R is --H, aryl or
substituted aryl, and Y.sub.1 and Y.sub.2 include but are not
limited to the following structures:
TABLE-US-00005 Structure Y.sub.1 Y.sub.2 III-97 C.sub.6H.sub.5 H
III-98 H C.sub.6H.sub.5 III-99 C.sub.6H.sub.5 C.sub.6H.sub.5
III-100 4-C.sub.6H.sub.4OH H III-101 H 4-C.sub.6H.sub.4OH III-102
3,4-C.sub.6H.sub.3(OH).sub.2 H III-103 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-104 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-105 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-106
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-107 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-108 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-109 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-110
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-111 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-112 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-113 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-114
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-115 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-116 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-117 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-118
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-119
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-120
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0172] Still further exemplary pteridines have the structure:
##STR00024##
wherein X.sub.1.dbd.NHR, wherein R is --H, aryl or substituted
aryl, and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00006 Structure Y.sub.1 Y.sub.2 III-121 C.sub.6H.sub.5 H
III-122 H C.sub.6H.sub.5 III-123 C.sub.6H.sub.5 C.sub.6H.sub.5
III-124 4-C.sub.6H.sub.4OH H III-125 H 4-C.sub.6H.sub.4OH III-126
3,4-C.sub.6H.sub.3(OH).sub.2 H III-127 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-128 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-129 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-130
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-131 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-132 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-133 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-134
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-135 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-136 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-137 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-138
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-139 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-140 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-141 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-142
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-143
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-144
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0173] Additional exemplary pteridines have the structure
##STR00025##
wherein X.sub.1.dbd.OR, wherein R is --H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00007 Structure Y.sub.1 Y.sub.2 III-145 C.sub.6H.sub.5 H
III-146 H C.sub.6H.sub.5 III-147 C.sub.6H.sub.5 C.sub.6H.sub.5
III-148 4-C.sub.6H.sub.4OH H III-149 H 4-C.sub.6H.sub.4OH III-150
3,4-C.sub.6H.sub.3(OH).sub.2 H III-151 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-152 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-153 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-154
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-155 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-156 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-157 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-158
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-159 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-160 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-161 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-162
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-163 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-164 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-165 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-166
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-167
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-168
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0174] Additional exemplary pteridines have the structure
##STR00026##
wherein X.sub.1.dbd.OR, wherein R is --H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00008 Structure Y.sub.1 Y.sub.2 III-169 C.sub.6H.sub.5 H
III-170 H C.sub.6H.sub.5 III-171 C.sub.6H.sub.5 C.sub.6H.sub.5
III-172 4-C.sub.6H.sub.4OH H III-173 H 4-C.sub.6H.sub.4OH III-174
3,4-C.sub.6H.sub.3(OH).sub.2 H III-175 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-176 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-177 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-178
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-179 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-180 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-181 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-182
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-183 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-184 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-185 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-186
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-187 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-188 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-189 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-190
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-191
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-192
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0175] In further embodiments, exemplary pteridines have the
structure:
##STR00027##
wherein X.sub.1.dbd.X.sub.2.dbd.Cl or NHR, wherein R is H,
(CH.sub.2).sub.2NHEt, (CH.sub.2).sub.3morpholyn-1-yl,
(CH.sub.2).sub.3(N-methylpiperazinyn-1-yl);
Y.sub.1.dbd.CH.sub.2glycinyl, CH.sub.2NHethoxy,
CH.sub.2NHCH.sub.2alkyl, CH.sub.2NHCH.sub.2t-Bu,
CH.sub.2NHCH.sub.2aryl, CH.sub.2NHCH.sub.2substituted aryl,
CH.sub.2NHCH.sub.2heteroaryl, CH.sub.2NHCH.sub.2substituted
heteroaryl with substituents being OH, and OMe, and Y.sub.2
includes but is not limited to the following structures:
TABLE-US-00009 Structure Y.sub.2 III-193 C.sub.6H.sub.5 III-194 H
III-195 4-C.sub.6H.sub.4OH III-196 3-C.sub.6H.sub.4OH III-197
2-C.sub.6H.sub.4OH III-198 naphthyl III-199 isonaphthyl III-200
4-tBuphenyl III-201 biphenyl III-202 2,3-di-methylphenyl III-203
fluorenyl III-204 oxophenyl III-205 thioindole III-206
C.sub.5H.sub.4N (pyr) III-207 4-C.sub.6H.sub.4F III-208
3-C.sub.6H.sub.4F III-209 4-C.sub.6H.sub.4OMe III-210
3-C.sub.6H.sub.4OMe III-211 2-C.sub.6H.sub.4OMe.
[0176] Additional exemplary compounds of structure (III) include
compounds having the structure:
##STR00028##
wherein X.sub.1.dbd.NHR, wherein R is H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00010 Structure Y.sub.1 Y.sub.2 III-212 C.sub.6H.sub.5 H
III-213 H C.sub.6H.sub.5 III-214 C.sub.6H.sub.5 C.sub.6H.sub.5
III-215 4-C.sub.6H.sub.4OH H III-216 H 4-C.sub.6H.sub.4OH III-217
3,4-C.sub.6H.sub.3(OH).sub.2 H III-218 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-219 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-220 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-221
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-222 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-223 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-224 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-225
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-226 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-227 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-228 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-229
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-230 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-231 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-232 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-233
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-234
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-235
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0177] Still further exemplary compounds of structure (III) include
the following:
##STR00029##
wherein X.sub.1.dbd.OR, wherein R is H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00011 Structure Y.sub.1 Y.sub.2 III-236 C.sub.6H.sub.5 H
III-237 H C.sub.6H.sub.5 III-238 C.sub.6H.sub.5 C.sub.6H.sub.5
III-239 4-C.sub.6H.sub.4OH H III-240 H 4-C.sub.6H.sub.4OH III-241
3,4-C.sub.6H.sub.3(OH).sub.2 H III-242 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-243 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-244 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-245
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-246 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-247 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-248 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-249
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-250 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-251 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-252 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-253
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-254 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-255 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-256 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-257
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-258
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-259
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0178] Compounds of structure (III) also include the following:
##STR00030##
wherein X.sub.1.dbd.NHR, wherein R is H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00012 Structure Y.sub.1 Y.sub.2 III-260 C.sub.6H.sub.5 H
III-261 H C.sub.6H.sub.5 III-262 C.sub.6H.sub.5 C.sub.6H.sub.5
III-263 4-C.sub.6H.sub.4OH H III-264 H 4-C.sub.6H.sub.4OH III-265
3,4-C.sub.6H.sub.3(OH).sub.2 H III-266 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-267 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-268 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-269
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-270 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-271 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-272 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-273
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-274 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-275 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-276 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-277
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-278 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-279 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-280 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-281
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-282
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-283
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0179] Still further exemplary compounds of structure (III)
include;
##STR00031##
wherein X.sub.1.dbd.OR, wherein R is H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00013 Structure Y.sub.1 Y.sub.2 III-284 C.sub.6H.sub.5 H
III-285 H C.sub.6H.sub.5 III-286 C.sub.6H.sub.5 C.sub.6H.sub.5
III-287 4-C.sub.6H.sub.4OH H III-288 H 4-C.sub.6H.sub.4OH III-289
3,4-C.sub.6H.sub.3(OH).sub.2 H III-290 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-291 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-292 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-293
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-294 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-295 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-296 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-297
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-298 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-299 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-300 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-301
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-302 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-303 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-304 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-305
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-306
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-307
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0180] Additional exemplary compounds of structure (III) include
quinoxalines having the structure:
##STR00032##
wherein X.sub.1.dbd.NHR and X.sub.2.dbd.NHR, wherein R is H, aryl
or substituted aryl, and Y.sub.1 and Y.sub.2 include but are not
limited to the following structures:
TABLE-US-00014 Structure Y.sub.1 Y.sub.2 III-308 C.sub.6H.sub.5 H
III-309 H C.sub.6H.sub.5 III-310 C.sub.6H.sub.5 C.sub.6H.sub.5
III-311 4-C.sub.6H.sub.4OH H III-312 H 4-C.sub.6H.sub.4OH III-313
3,4-C.sub.6H.sub.3(OH).sub.2 H III-314 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-315 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-316 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-317
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-318 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-319 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-320 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-321
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-322 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-323 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-324 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-325
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-326 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-327 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-328 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-329
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-330
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-331
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0181] Additional quinoxalines contemplated for use in the practice
of the invention include the following:
##STR00033##
wherein X.sub.1.dbd.X.sub.2.dbd.OR, wherein R is --H, aryl or
substituted aryl, and Y.sub.1 and Y.sub.2 include but are not
limited to the following structures:
TABLE-US-00015 Structure Y.sub.1 Y.sub.2 III-332 C.sub.6H.sub.5 H
III-333 H C.sub.6H.sub.5 III-334 C.sub.6H.sub.5 C.sub.6H.sub.5
III-335 4-C.sub.6H.sub.4OH H III-336 H 4-C.sub.6H.sub.4OH III-337
3,4-C.sub.6H.sub.3(OH).sub.2 H III-338 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-339 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-340 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-341
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-342 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-343 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-344 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-345
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-346 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-347 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-348 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-349
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-350 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-351 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-352 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-353
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-354
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-355
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0182] Still further exemplary quinoxalines include:
##STR00034##
wherein when X.sub.1.dbd.OR and X.sub.2.dbd.NHR, wherein R is H,
aryl or substituted aryl, and Y.sub.1 and Y.sub.2 include but are
not limited to the following structures:
TABLE-US-00016 Structure Y.sub.1 Y.sub.2 III-356 C.sub.6H.sub.5 H
III-357 H C.sub.6H.sub.5 III-358 C.sub.6H.sub.5 C.sub.6H.sub.5
III-359 4-C.sub.6H.sub.4OH H III-360 H 4-C.sub.6H.sub.4OH III-361
3,4-C.sub.6H.sub.3(OH).sub.2 H III-362 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-363 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-364 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-365
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-366 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-367 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-368 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-369
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-370 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-371 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-372 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-373
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-374 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-375 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-376 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-377
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-378
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-379
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0183] Additional exemplary quinoxalines have the structure:
##STR00035##
wherein X.sub.1.dbd.NHR and X.sub.2.dbd.OR, wherein R is H, aryl or
substituted aryl, and Y.sub.1 and Y.sub.2 include but are not
limited to the following structures:
TABLE-US-00017 Structure Y.sub.1 Y.sub.2 III-380 C.sub.6H.sub.5 H
III-381 H C.sub.6H.sub.5 III-382 C.sub.6H.sub.5 C.sub.6H.sub.5
III-383 4-C.sub.6H.sub.4OH H III-384 H 4-C.sub.6H.sub.4OH III-385
3,4-C.sub.6H.sub.3(OH).sub.2 H III-386 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-387 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-388 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-389
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-390 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-391 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-392 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-393
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-394 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-395 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-396 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-397
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-398 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-399 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-400 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-401
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-402
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-403
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0184] Still further exemplary quinoxalines have the structure:
##STR00036##
wherein X.sub.1.dbd.NHR, wherein R is H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00018 Structure Y.sub.1 Y.sub.2 III-404 C.sub.6H.sub.5 H
III-405 H C.sub.6H.sub.5 III-406 C.sub.6H.sub.5 C.sub.6H.sub.5
III-407 4-C.sub.6H.sub.4OH H III-408 H 4-C.sub.6H.sub.4OH III-409
3,4-C.sub.6H.sub.3(OH).sub.2 H III-410 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-411 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-412 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-413
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-414 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-415 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-416 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-417
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-418 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-419 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-420 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-421
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-422 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-423 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-424 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-425
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-426
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-427
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0185] Additional exemplary quinoxalines have the structure:
##STR00037##
wherein X.sub.1.dbd.NHR, wherein R is H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00019 Structure Y.sub.1 Y.sub.2 III-428 C.sub.6H.sub.5 H
III-429 H C.sub.6H.sub.5 III-430 C.sub.6H.sub.5 C.sub.6H.sub.5
III-431 4-C.sub.6H.sub.4OH H III-432 H 4-C.sub.6H.sub.4OH III-433
3,4-C.sub.6H.sub.3(OH).sub.2 H III-434 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-435 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-436 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-437
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-438 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-439 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-440 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-441
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-442 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-443 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-444 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-445
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-446 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-447 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-448 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-449
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-450
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-451
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0186] Still further exemplary quinoxalines have the structure:
##STR00038##
wherein X.sub.1.dbd.OR, wherein R is H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00020 Structure Y.sub.1 Y.sub.2 III-452 C.sub.6H.sub.5 H
III-453 H C.sub.6H.sub.5 III-454 C.sub.6H.sub.5 C.sub.6H.sub.5
III-455 4-C.sub.6H.sub.4OH H III-456 H 4-C.sub.6H.sub.4OH III-457
3,4-C.sub.6H.sub.3(OH).sub.2 H III-458 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-459 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-460 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-461
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-462 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-463 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-464 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-465
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-466 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-467 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-468 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-469
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-470 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-471 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-472 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-473
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-474
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-475
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0187] Further exemplary quinoxalines have the structure:
##STR00039##
wherein X.sub.1.dbd.OR, wherein R is H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00021 Structure Y.sub.1 Y.sub.2 III-476 C.sub.6H.sub.5 H
III-477 H C.sub.6H.sub.5 III-478 C.sub.6H.sub.5 C.sub.6H.sub.5
III-479 4-C.sub.6H.sub.4OH H III-480 H 4-C.sub.6H.sub.4OH III-481
3,4-C.sub.6H.sub.3(OH).sub.2 H III-482 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-483 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-484 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-485
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-486 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-487 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-488 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-489
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-490 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-491 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-492 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-493
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-494 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-495 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-496 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-497
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-498
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-499
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0188] Still further exemplary compounds of structure (III)
include:
##STR00040##
wherein X.sub.1.dbd.NHR, wherein R is H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00022 Structure Y.sub.1 Y.sub.2 III-500 C.sub.6H.sub.5 H
III-501 H C.sub.6H.sub.5 III-502 C.sub.6H.sub.5 C.sub.6H.sub.5
III-503 4-C.sub.6H.sub.4OH H III-504 H 4-C.sub.6H.sub.4OH III-505
3,4-C.sub.6H.sub.3(OH).sub.2 H III-506 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-507 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-508 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-509
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-510 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-511 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-512 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-513
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-514 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-515 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-516 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-517
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-518 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-519 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-520 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-521
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-522
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-523
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0189] Additional compounds of structure (III) include the
following:
##STR00041##
wherein X.sub.1.dbd.OR, wherein R is H, aryl or substituted aryl,
and Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00023 Structure Y.sub.1 Y.sub.2 III-524 C.sub.6H.sub.5 H
III-525 H C.sub.6H.sub.5 III-526 C.sub.6H.sub.5 C.sub.6H.sub.5
III-527 4-C.sub.6H.sub.4OH H III-528 H 4-C.sub.6H.sub.4OH III-529
3,4-C.sub.6H.sub.3(OH).sub.2 H III-530 H
3,4-C.sub.6H.sub.3(OH).sub.2 III-531 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-532 C.sub.6H.sub.5 4-C.sub.6H.sub.4F III-533
4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-534 C.sub.6H.sub.5
4-C.sub.6H.sub.4Br III-535 4-C.sub.6H.sub.4OPh C.sub.6H.sub.5
III-536 C.sub.6H.sub.5 4-C.sub.6H.sub.4OPh III-537
4-C.sub.6H.sub.4OH C.sub.6H.sub.5 III-538 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-539 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-540 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-541
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-542 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-543 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-544 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-545
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-546
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-547
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0190] Still further exemplary compounds of structure (III)
include:
##STR00042##
wherein Y.sub.1 and Y.sub.2 include but are not limited to the
following structures:
TABLE-US-00024 Structure Y.sub.1 Y.sub.2 III-547 C.sub.6H.sub.5 H
III-548 C.sub.6H.sub.5 C.sub.6H.sub.5 III-549 4-C.sub.6H.sub.4OH H
III-550 3,4-C.sub.6H.sub.3(OH).sub.2 H III-551 4-C.sub.6H.sub.4F
C.sub.6H.sub.5 III-552 4-C.sub.6H.sub.4Br C.sub.6H.sub.5 III-553
4-C.sub.6H.sub.4OPh C.sub.6H.sub.5 III-554 C.sub.6H.sub.5
4-C.sub.6H.sub.4OH III-555 C.sub.5H.sub.4N (pyr) C.sub.5H.sub.4N
(pyr) III-556 4-C.sub.6H.sub.4F 4-C.sub.6H.sub.4F III-557
3-C.sub.6H.sub.4F 3-C.sub.6H.sub.4F III-558 4-C.sub.6H.sub.4OMe
4-C.sub.6H.sub.4OMe III-559 3-C.sub.6H.sub.4OMe 3-C.sub.6H.sub.4OMe
III-560 4-C.sub.6H.sub.4OH 4-C.sub.6H.sub.4OH III-561
3-C.sub.6H.sub.4OH 3-C.sub.6H.sub.4OH III-562
3,4-C.sub.6H.sub.3(OH).sub.2 3,4-C.sub.6H.sub.3(OH).sub.2 III-563
Y.sub.1 and Y.sub.2 taken together to form a phenathrolinyl
group.
[0191] Additional exemplary compounds of structure (III)
include:
##STR00043##
wherein X.sub.1.dbd.NHR, wherein R is H, aryl, substituted aryl, or
aroyl, Y.sub.1.dbd.NHR, or R,
[0192] wherein R.dbd.H, alkyl or branched alkyl, and Y.sub.2
includes but is not limited to the following structures:
TABLE-US-00025 Structure Y.sub.2 III-564 C.sub.6H.sub.5 III-565 H
III-566 4-C.sub.6H.sub.4OH III-567 3-C.sub.6H.sub.4OH III-568
2-C.sub.6H.sub.4OH III-569 naphthyl III-570 isonaphthyl III-571
4-tBuphenyl III-572 biphenyl III-573 2,3-diMephenyl III-574
fluorenyl III-575 oxophenyl III-576 thioindole III-577
C.sub.5H.sub.4N (pyr) III-578 4-C.sub.6H.sub.4F III-579
3-C.sub.6H.sub.4F III-580 4-C.sub.6H.sub.4OMe III-581
3-C.sub.6H.sub.4OMe III-582 2-C.sub.6H.sub.4OMe
[0193] Still further exemplary compounds of structure (III) include
asymmetric triazines, such as:
##STR00044##
wherein Y.sub.1.dbd.NHR or R, wherein R.dbd.H, alkyl or branched
alkyl, and Y.sub.2 includes but is not limited to the following
structures:
TABLE-US-00026 Structure Y.sub.2 III-583 C.sub.6H.sub.5 III-584 H
III-585 4-C.sub.6H.sub.4OH III-586 3-C.sub.6H.sub.4OH III-587
2-C.sub.6H.sub.4OH III-588 naphthyl III-589 isonaphthyl III-590
4-tBuphenyl III-591 biphenyl III-592 2,3-diMephenyl III-593
fluorenyl III-594 oxophenyl III-595 thioindole III-596
C.sub.5H.sub.4N (pyr) III-597 4-C.sub.6H.sub.4F III-598
3-C.sub.6H.sub.4F III-599 4-C.sub.6H.sub.4OMe III-600
3-C.sub.6H.sub.4OMe III-601 2-C.sub.6H.sub.4OMe
[0194] In yet another embodiment of the invention, compounds are
provided having structure (IV):
##STR00045## [0195] wherein: [0196] L is an arylene, substituted
arylene, oxyarylene, or substituted oxyarylene linking moiety,
[0197] C is 5- or 6-membered aromatic or heteroaromatic ring,
[0198] each X is independently OR, NR.sub.2, or SR, wherein R is H
or lower alkyl, [0199] Z.sub.1-Z.sub.4 are each independently CH or
N, and [0200] m is 1 to 4.
[0201] In some embodiments, the linking moiety L is an arylene
moiety, and Z is N, as exemplified by the following structures:
##STR00046##
wherein, Z.dbd.N or CH, X.sub.1.dbd.H or OH, and
X.sub.2.dbd.NH.sub.2 or OH.
[0202] In another embodiment, the linking moiety L is an oxyarylene
moiety, as exemplified by the following structures:
##STR00047##
wherein, Z.dbd.N or CH, X.sub.1.dbd.H or OH, and
X.sub.2.dbd.NH.sub.2 or OH.
[0203] In still another embodiment, compounds are provided having
the structure (V):
##STR00048## [0204] wherein: [0205] R.sub.1, x, and y are as
defined above, [0206] R.sub.3 is --H, --SO.sub.3H, or
--SO.sub.2NMe.sub.2, [0207] M is NH, CO, SO.sub.2,
(CH.sub.2).sub.p, wherein p is 0 to 2, [0208] G is aryl or
heteroaryl, and [0209] x and y are each independently 0-4. In an
additional embodiment, there are provided bis-pteridine compounds.
An exemplary bis-pteridine compound according to the invention has
the structure:
##STR00049##
[0210] As used herein, the term "heterocyclic", when used to
describe an aromatic ring, means that the aromatic ring contains at
least one heteroatom. As used herein, the term "heteroatom" refers
to N, O, S, and the like.
[0211] As used herein, the term "alkyl" refers to a monovalent
straight or branched chain hydrocarbon group having from one to
about 12 carbon atoms, including methyl, ethyl, n-propyl,
isopropyl, n-butyl, isobutyl, tert-butyl, n-hexyl, and the
like.
[0212] As used herein, "substituted alkyl" refers to alkyl groups
further bearing one or more substituents selected from hydroxy,
alkoxy, mercapto, cycloalkyl, substituted cycloalkyl, heterocyclic,
substituted heterocyclic, aryl, substituted aryl, heteroaryl,
substituted heteroaryl, aryloxy, substituted aryloxy, halogen,
cyano, nitro, amino, amido, --C(O)H, acyl, oxyacyl, carboxyl,
sulfonyl, sulfonamide, sulfuryl, and the like.
[0213] As used herein, "lower alkyl" refers to alkyl groups having
from 1 to about 6 carbon atoms.
[0214] As used herein, "alkenyl" refers to straight or branched
chain hydrocarbyl groups having one or more carbon-carbon double
bonds, and having in the range of about 2 up to 12 carbon atoms,
and "substituted alkenyl" refers to alkenyl groups further bearing
one or more substituents as set forth above.
[0215] As used herein, "alkynyl" refers to straight or branched
chain hydrocarbyl groups having at least one carbon-carbon triple
bond, and having in the range of about 2 up to 12 carbon atoms, and
"substituted alkynyl" refers to alkynyl groups further bearing one
or more substituents as set forth above.
[0216] As used herein, "aryl" refers to aromatic groups having in
the range of 6 up to 14 carbon atoms and "substituted aryl" refers
to aryl groups further bearing one or more substituents as set
forth above.
[0217] As used herein, "heteroaryl" refers to aromatic rings
containing one or more heteroatoms (e.g., N, O, S, or the like) as
part of the ring structure, and having in the range of 3 up to 14
carbon atoms and "substituted heteroaryl" refers to heteroaryl
groups further bearing one or more substituents as set forth
above.
[0218] As used herein, "alkoxy" refers to the moiety --O-alkyl-,
wherein alkyl is as defined above, and "substituted alkoxy" refers
to alkoxyl groups further bearing one or more substituents as set
forth above.
[0219] As used herein, "cycloalkyl" refers to ring-containing alkyl
groups containing in the range of about 3 up to 8 carbon atoms, and
"substituted cycloalkyl" refers to cycloalkyl groups further
bearing one or more substituents as set forth above.
[0220] As used herein, "heterocyclic", when not used with reference
to an aromatic ring, refers to cyclic (i.e., ring-containing)
groups containing one or more heteroatoms (e.g., N, O, S, or the
like) as part of the ring structure, and having in the range of 3
up to 14 carbon atoms and "substituted heterocyclic" refers to
heterocyclic groups further bearing one or more substituents as set
forth above.
[0221] As used herein, "alkylaryl" refers to alkyl-substituted aryl
groups and "substituted alkylaryl" refers to alkylaryl groups
further bearing one or more substituents as set forth above.
[0222] As used herein, "arylalkyl" refers to aryl-substituted alkyl
groups and "substituted arylalkyl" refers to arylalkyl groups
further bearing one or more substituents as set forth above.
[0223] As used herein, "arylalkenyl" refers to aryl-substituted
alkenyl groups and "substituted arylalkenyl" refers to arylalkenyl
groups further bearing one or more substituents as set forth
above.
[0224] As used herein, "arylalkynyl" refers to aryl-substituted
alkynyl groups and "substituted arylalkynyl" refers to arylalkynyl
groups further bearing one or more substituents as set forth
above.
[0225] As used herein, divalent aromatic groups having in the range
of 6 up to 14 carbon atoms and "substituted arylene" refers to
arylene groups further bearing one or more substituents as set
forth above.
[0226] As used herein, "oxyarylene" refers to the moiety
"O-arylene", wherein arylene is as defined above and "substituted
oxyarylene" refers to oxyarylene groups further bearing one or more
substituents as set forth above.
[0227] Invention compounds can be prepared by a variety of methods
well-known to those skilled in the art. For example, Scheme A
illustrates three exemplary syntheses for invention compounds of
structure (I).
##STR00050## ##STR00051##
[0228] Scheme B illustrates an exemplary synthesis for invention
compounds of structure (II).
##STR00052##
[0229] Scheme C illustrates two of several exemplary syntheses for
invention compounds of structure (III).
##STR00053##
[0230] Scheme D illustrates an exemplary synthesis for invention
compounds of structure (IV).
##STR00054##
[0231] Scheme E below illustrates an exemplary synthesis for
compounds of structure (V).
##STR00055##
[0232] In a further embodiment of the invention, there provided
methods for treating a disorder, comprising administering to a
subject in need thereof an effective amount of a compound having
the structure (VI):
##STR00056## [0233] wherein: [0234] A and B are each independently
5- or 6-membered aromatic rings, wherein at least one of A and B is
an aromatic heterocyclic ring having at least one heteroatom in the
heterocyclic ring, [0235] each X is independently OR, NR.sub.2, or
SR, wherein R is H or lower alkyl, [0236] each Y is independently
hydrogen, alkyl, substituted alkyl, alkenyl substituted alkenyl,
alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,
heterocyclic, substituted heterocyclic, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, alkylaryl, substituted
alkylaryl, arylalkyl, substituted arylalkyl, arylalkenyl,
substituted arylalkenyl, arylalkynyl, substituted arylalkynyl,
aroyl, substituted aroyl, acyl, or substituted acyl, with the
proviso that at least one Y is not hydrogen, or [0237] when n is 2,
each Y is taken together to form a fused aromatic ring system
comprising at least one aromatic ring, and [0238] m and n are each
independently 1 to 4, thereby treating the disorder.
[0239] Rings A and B taken together may form a variety of fused
aromatic heterocyclic groups suitable for use in the practice of
the present invention. For example, rings A and B taken together
may form aromatic heterocycles such as quinoxaline, pteridine,
benzoxazine, benzoxazole, benzimidazole, 1,2-benzodiazole, indole,
isoindole, quinoline, isoquinoline, phthalazine, naphthyridine,
quinazoline, cinnoline, purine, benzothiazole, benzofuran,
isobenzofuran, benzothiophene, chromene, and the like. In one
embodiment, rings A and B taken together form a quinoxaline. In a
further embodiment, rings A and B taken together form a pteridine.
In a still further embodiment, rings A and B taken together form a
benzimidazole.
[0240] Quinoxalines contemplated for use in the methods of the
present invention have the structure:
##STR00057## [0241] wherein: [0242] each X is independently H, OR,
NR.sub.2, or SR, wherein R is H or lower alkyl, [0243] each Y is
independently hydrogen, alkyl, substituted alkyl, alkenyl
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, heterocyclic, substituted heterocyclic,
aryl, substituted aryl, heteroaryl, substituted heteroaryl,
alkylaryl, substituted alkylaryl, arylalkyl, substituted arylalkyl,
arylalkenyl, substituted arylalkenyl, arylalkynyl, substituted
arylalkynyl, with the proviso that at least one Y is not hydrogen,
or [0244] when n is 2, each Y is taken together to form a fused
aromatic ring system comprising at least one aromatic ring, [0245]
m is 1 to 4, and [0246] n is 1 or 2.
[0247] In one embodiment, quinoxalines contemplated for use in the
methods of the present invention have the structure:
##STR00058## [0248] wherein: [0249] X is OR, NR.sub.2, or SR,
wherein R is H or lower alkyl, [0250] Y is aryl, substituted aryl,
heteroaryl, or substituted heteroaryl, and [0251] n is 1 or 2.
[0252] Pteridines contemplated for use in the methods of the
present invention have the structure:
##STR00059## [0253] wherein: [0254] each X is independently H, OR,
NR.sub.2, or SR, wherein R is H or lower alkyl, [0255] each Y is
independently hydrogen, alkyl, substituted alkyl, alkenyl
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, heterocyclic, substituted heterocyclic,
aryl, substituted aryl, heteroaryl, substituted heteroaryl,
alkylaryl, substituted alkylaryl, arylalkyl, substituted arylalkyl,
arylalkenyl, substituted arylalkenyl, arylalkynyl, substituted
arylalkynyl, aroyl, substituted aroyl, acyl, or substituted acyl,
with the proviso that at least one Y is not hydrogen, or [0256]
when n is 2, each Y is taken together to form a fused aromatic ring
system comprising at least one aromatic ring, and [0257] m and n
are each independently 1 or 2.
[0258] In one embodiment, pteridines contemplated for use in the
methods of the present invention have the structure:
##STR00060## [0259] wherein: [0260] X is OR, NR.sub.2, or SR,
wherein R is H or lower alkyl, [0261] Y is aryl, substituted aryl,
heteroaryl, or substituted heteroaryl, and [0262] n is 1 or 2.
[0263] Benzimidazoles, oxazoles, or thiazoles contemplated for use
in the methods of the present invention have the structure:
##STR00061## [0264] wherein: [0265] Z is N, O, or S, [0266] each X
is independently H, OR, NR.sub.2, or SR, wherein R is H or lower
alkyl, [0267] each Y is independently hydrogen, alkyl, substituted
alkyl, alkenyl substituted alkenyl, alkynyl, substituted alkynyl,
cycloalkyl, substituted cycloalkyl, heterocyclic, substituted
heterocyclic, aryl, substituted aryl, heteroaryl, substituted
heteroaryl, alkylaryl, substituted alkylaryl, arylalkyl,
substituted arylalkyl, arylalkenyl, substituted arylalkenyl,
arylalkynyl, substituted arylalkynyl, aroyl, substituted aroyl,
acyl, or substituted acyl, with the proviso that at least one Y is
not hydrogen, or [0268] when n is 2, each Y is taken together to
form a fused aromatic ring system comprising at least one aromatic
ring, and [0269] m is 1 to 4, and [0270] n is 1 or 2.
[0271] In one embodiment, benzimidazoles contemplated for use in
the methods of the present invention have the structure:
##STR00062## [0272] wherein: [0273] each X is independently H, OR,
NR.sub.2, or SR, wherein R is H or lower alkyl, [0274] Y is aryl,
substituted aryl, heteroaryl, or substituted heteroaryl, and [0275]
m is 1-4.
[0276] In a further embodiment of the invention, there are provided
methods for treating a disorder such as those associated with
vascular permeability and/or angiogenesis and/or other aspects of
compromised vasculostasis including administering to a subject in
need thereof an effective amount of a compound having structure
(VII):
##STR00063## [0277] wherein: [0278] A, B, C, and D are each
independently C, N, O, or S, [0279] each X is independently H, OR,
NR.sub.2, or SR, wherein R is H or lower alkyl, [0280] each Y is
independently hydrogen, alkyl, substituted alkyl, alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, heterocyclic, substituted heterocyclic,
aryl, substituted aryl, heteroaryl, substituted heteroaryl,
alkylaryl, substituted alkylaryl, arylalkyl, substituted arylalkyl,
arylalkenyl, substituted arylalkenyl, arylalkynyl, substituted
arylalkynyl, aroyl, substituted aroyl, acyl, or substituted acyl,
with the proviso that at least one Y is not hydrogen, and [0281] m
and n are each independently 1 to 4, thereby treating the
disorder.
[0282] In one aspect of this embodiment, the compound has the
structure:
##STR00064## [0283] wherein: [0284] each X is independently H, OR,
NR.sub.2, or SR, wherein R is H or lower alkyl, [0285] each Y is
independently aryl or substituted aryl, [0286] m is 1 or 2, and
[0287] n is 1-4.
[0288] In a further aspect of this embodiment, the compound has the
structure:
##STR00065##
[0289] In one embodiment, the present invention is based on the
discovery that a combination therapy including interleukin-2 (IL-2)
and chemical compounds described herein, some of which are
effective kinase inhibitors, administered during IL-2 therapy,
mitigates or lessens the adverse effects of IL-2. While not wanting
to be bound by a particular theory, it is likely that the effect
occurs while preserving or enhancing the beneficial effect of IL-2
such that the disease or disorder is treated. While IL-2 is
described in the present application as an illustrative example, it
should be understood that the invention includes combination
therapy including a compound of the invention, including but not
limited to vasculostatic agents, such as tyrosine, serine or
threonine kinase inhibitors, for example, Src-family inhibitors,
and immunomodulatory molecules. In particular, such
immunomodulatory molecules include those that result in vascular
leakage. Cytokines, and in particular IL-2, are examples of such
immunomodulatory molecules.
[0290] Such inhibitors, in combination with IL-2, are effective in
blocking vascular leakage typically associated with IL-2
administration. Thus, compositions and methods are provided for
treating disorders associated with VLS. In one embodiment, the
invention provides a composition containing a therapeutically
effective amount of IL-2 and a vasculostatic agent or compound as
described herein in a pharmaceutically acceptable carrier.
[0291] Some of the compounds are kinase inhibitors, such as
Src-family tyrosine kinases, and therefore are useful in treating a
wide variety of disorders resulting from aberrant kinase activity,
in addition to treating disorders associates with IL-2
administration. Kinase-associated disorders are those disorders
which result from aberrant kinase activity, and/or which are
alleviated by the inhibition of one or more enzymes within a kinase
family. For example, Lck inhibitors are of value in the treatment
of a number of such disorders (e.g., the treatment of autoimmune
diseases), as Lck inhibition blocks T cell activation. Similarly,
Src family inhibitors are of value in treating a variety of cancers
as Src inhibition impacts tumor cell invasion, metastases and
survival.
[0292] The compounds and methods of the present invention, either
when administered alone or in combination with other agents
described herein (e.g., chemotherapeutic agents or protein
therapeutic agents) are useful in treating a variety of disorders
associated with compromised vasculostasis including but not limited
to, for example: stroke, cardiovascular disease, myocardial
infarction, congestive heart failure, cardiomyopathy, myocarditis,
ischemic heart disease, coronary artery disease, cardiogenic shock,
vascular shock, pulmonary hypertension, pulmonary edema (including
cardiogenic pulmonary edema), cancer, pleural effusions, rheumatoid
arthritis, diabetic retinopathy, retinitis pigmentosa, and
retinopathies, including diabetic retinopathy and retinopathy of
prematurity, inflammatory diseases, restenosis, edema (including
edema associated with pathologic situations such as cancers and
edema induced by medical interventions such as chemotherapy),
asthma, acute or adult respiratory distress syndrome (ARDS), lupus,
vascular leakage, transplant (such as organ transplant, acute
transplant or heterograft or homograft (such as is employed in burn
treatment)) rejection; protection from ischemic or reperfusion
injury such as ischemic or reperfusion injury incurred during organ
transplantation, transplantation tolerance induction; ischemic or
reperfusion injury following angioplasty; arthritis (such as
rheumatoid arthritis, psoriatic arthritis or osteoarthritis);
multiple sclerosis; inflammatory bowel disease, including
ulcerative colitis and Crohn's disease; lupus (systemic lupus
crythematosis); graft vs. host diseases; T-cell mediated
hypersensitivity diseases, including contact hypersensitivity,
delayed-type hypersensitivity, and gluten-sensitive enteropathy
(Celiac disease); Type 1 diabetes; psoriasis; contact dermatitis
(including that due to poison ivy); Hashimoto's thyroiditis;
Sjogren's syndrome; Autoimmune Hyperthyroidism, such as Graves'
disease; Addison's disease (autoimmune disease of the adrenal
glands); autoimmune polyglandular disease (also known as autoimmune
polyglandular syndrome); autoimmune alopecia; pernicious anemia;
vitiligo; autoimmune hypopituatarism; Guillain-Barre syndrome;
other autoimmune diseases; cancers, including those where kinases
such as Src-family kinases are activated or overexpressed, such as
colon carcinoma and thymoma, or cancers where kinase activity
facilitates tumor growth or survival; glomerulonephritis, serum
sickness; uticaria; allergic diseases such as respiratory allergies
(asthma, hayfever, allergic rhinitis) or skin allergies; mycosis
fungoides; acute inflammatory responses (such as acute or adult
respiratory distress syndrome and ischemia/reperfusion injury);
dermatomyositis; alopecia greata; chronic actinic dermatitis;
eczema; Behcet's disease; Pustulosis palmoplanteris; Pyoderma
gangrenum; Sezary's syndrome; atopic dermatitis; systemic
schlerosis; morphea; peripheral limb ischemia and ischemic limb
disease; bone disease such as osteoporosis, osteomalacia,
hyperparathyroidism, Paget's disease, and renal osteodystrophy;
vascular leak syndromes, including vascular leak syndromes induced
by chemotherapies or immunomodulators such as IL-2; spinal cord and
brain injury or trauma; glaucoma; retinal diseases, including
macular degeneration; vitreoretinal disease; pancreatitis;
vasculatides, including vasculitis, Kawasaki disease,
thromboangiitis obliterans, Wegener's granulomatosis, and Behcet's
disease; scleroderma; preeclampsia; thalassemia; Kaposi's sarcoma;
von Hippel Lindau disease; and the like.
[0293] "Treating cancer" as used herein refers to providing a
therapeutic benefit to the cancer patient, e.g. the therapy extends
the mean survival time of patients, increases the percentage of
patients surviving at a given timepoint, extends the mean time to
disease progression, reduces or stabilizes tumor burden or improves
quality of life for the patient or any of the above, for example.
While not wanting to be bound by a particular theory, some of the
compounds of the invention may be cytostatic and therefore have
activity directly on the tumor cells.
[0294] As used herein, "kinase" refers to any enzyme that catalyze
the addition of phosphate groups to a protein residue, for example
serine and threonine kinases catalyze the addition of phosphate
groups to serine and threonine residues.
[0295] As used herein, the terms "Src kinase" or "Src kinase
family" or "Src family" refer to the related homologs or analogs
belonging to the mammalian family of Src kinases, including, for
example, the widely expressed c-Src, Fyn, Yes and Lyn kinases and
the hematopoietic-restricted kinases Hck, Fgr, Lck and Blk. As used
herein, the terms "Src kinase signaling pathway" or "Src cascade"
refer to both the upstream and downstream components of the Src
signaling cascade.
[0296] Src-family tyrosine kinases other than Lck, such as Hck and
Fgr, are important in the Fc gamma receptor induced respiratory
burst of neutrophils as well as the Fc gamma receptor responses of
monocytes and macrophages. The compositions and methods of the
present invention may be useful in inhibiting the Fc gamma induced
respiratory burst response in neutrophils, and may also be useful
in inhibiting the Fc gamma dependent production of TNF alpha. The
ability to inhibit Fc gamma receptor dependent neutrophil, monocyte
and macrophage responses would result in additional
anti-inflammatory activity for the compounds employed in invention
methods. This activity would be especially of value, for example,
in the treatment of inflammatory diseases, such as arthritis or
inflammatory bowel disease. The compositions and methods of the
present invention may also be useful in the treatment of autoimmune
glomerulonephritis and other instances of glomerulonephritis
induced by deposition of immune complexes in the kidney that
trigger Fc gamma receptor responses and which can lead to kidney
damage.
[0297] In addition, certain Src-family tyrosine kinases, such as
Lyn and Src, may be important in the Fc epsilon receptor induced
degranulation of mast cells and basophils that plays an important
role in asthma, allergic rhinitis, and other allergic disease. Fc
epsilon receptors are stimulated by IgE-antigen complexes.
Compounds employed in the methods of the present invention may
inhibit the Fc epsilon induced degranulation responses. The ability
to inhibit Fc epsilon receptor dependent mast cell and basophil
responses may result in additional anti-inflammatory activity for
the present compounds beyond their effect on T cells.
[0298] The present invention also provides articles of manufacture
comprising packaging material and a pharmaceutical composition
contained within said packaging material, wherein said packaging
material comprises a label which indicates that said pharmaceutical
composition can be used for treatment of disorders and wherein said
pharmaceutical composition comprises a compound according to the
present invention. Thus, in one aspect, the invention provides a
pharmaceutical composition including both a therapeutic and a
compound of the invention (e.g, as shown in FIG. 1), wherein the
compound is present in a concentration effective to reduce vascular
leakage associated with indications or therapeutics which have
vascular leak as a side-effect. For example, administration of a
compound of the invention in conjunction with IL-2, immunotoxins,
antibodies or chemotherapeutics. In these cases, IL-2, immunotoxin,
antibody or chemotherapeutic concentration can be determined by one
of skill in the art according to standard treatment regimen or as
determined by an in vivo animal assay, for example.
[0299] The present invention also provides pharmaceutical
compositions comprising IL-2, immunotoxin, antibody or
chemotherapeutic and at least one invention compound in an amount
effective for inhibiting vascular permeability, and a
pharmaceutically acceptable vehicle or diluent. The compositions of
the present invention may contain other therapeutic agents as
described below, and may be formulated, for example, by employing
conventional solid or liquid vehicles or diluents, as well as
pharmaceutical additives of a type appropriate to the mode of
desired administration (for example, excipients, binders,
preservatives, stabilizers, flavors, etc.) according to techniques
such as those well known in the art of pharmaceutical
formulation.
[0300] The compounds of the invention may be formulated into
therapeutic compositions as natural or salt forms. Pharmaceutically
acceptable non-toxic salts include the base addition salts (formed
with free carboxyl or other anionic groups) which may be derived
from inorganic bases such as, for example, sodium, potassium,
ammonium, calcium, or ferric hydroxides, and such organic bases as
isopropylamine, trimethylamine, 2-ethylamino-ethanol, histidine,
procaine, and the like. Such salts may also be formed as acid
addition salts with any free cationic groups and will generally be
formed with inorganic acids such as, for example, hydrochloric,
sulfuric, or phosphoric acids, or organic acids such as acetic,
citric, p-toluenesulfonic, methanesulfonic acid, oxalic, tartaric,
mandelic, and the like. Salts of the invention include amine salts
formed by the protonation of an amino group with inorganic acids
such as hydrochloric acid, hydrobromic acid, hydroiodic acid,
sulfuric acid, phosphoric acid, and the like. Salts of the
invention also include amine salts formed by the protonation of an
amino group with suitable organic acids, such as p-toluenesulfonic
acid, acetic acid, and the like. Additional excipients which are
contemplated for use in the practice of the present invention are
those available to those of ordinary skill in the art, for example,
those found in the United States Pharmacopeia Vol. XXII and
National Formulary Vol. XVII, U.S. Pharmacopeia Convention, Inc.,
Rockville, Md. (1989), the relevant contents of which is
incorporated herein by reference. In addition, polymorphs of the
invention compounds are included in the present invention.
[0301] Invention pharmaceutical compositions may be administered by
any suitable means, for example, orally, such as in the form of
tablets, capsules, granules or powders; sublingually; buccally;
parenterally, such as by subcutaneous, intravenous, intramuscular,
intrathecal, or intracisternal injection or infusion techniques
(e.g., as sterile injectable aqueous or non-aqueous solutions or
suspensions); nasally such as by inhalation spray; topically, such
as in the form of a cream or ointment; or rectally such as in the
form of suppositories; in dosage unit formulations containing
non-toxic, pharmaceutically acceptable vehicles or diluents. The
present compounds may, for example, be administered in a form
suitable for immediate release or extended release. Immediate
release or extended release may be achieved by the use of suitable
pharmaceutical compositions comprising the present compounds, or,
particularly in the case of extended release, by the use of devices
such as subcutaneous implants or osmotic pumps. The present
compounds may also be administered liposomally.
[0302] In addition to primates, such as humans, a variety of other
mammals can be treated according to the method of the present
invention. For instance, mammals including, but not limited to,
cows, sheep, goats, horses, dogs, cats, guinea pigs, rats or other
bovine, ovine, equine, canine, feline, rodent or murine species can
be treated. However, the method can also be practiced in other
species, such as avian species (e.g., chickens).
[0303] The term "therapeutically effective amount" means the amount
of the compound or pharmaceutical composition that will elicit the
biological or medical response of a tissue, system, animal or human
that is being sought by the researcher, veterinarian, medical
doctor or other clinician, e.g., restoration or maintenance of
vasculostasis or prevention of the compromise or loss or
vasculostasis; reduction of tumor burden; reduction of morbidity
and/or mortality.
[0304] By "pharmaceutically acceptable" it is meant the carrier,
diluent or excipient must be compatible with the other ingredients
of the formulation and not deleterious to the recipient
thereof.
[0305] The terms "administration of" and or "administering a"
compound should be understood to mean providing a compound of the
invention or pharmaceutical composition to the subject in need of
treatment.
[0306] The pharmaceutical compositions for the administration of
the compounds of this embodiment either alone or in combination
with IL-2, immunotoxin, antibody or chemotherapeutic may
conveniently be presented in dosage unit form and may be prepared
by any of the methods well known in the art of pharmacy. All
methods include the step of bringing the active ingredient into
association with the carrier which constitutes one or more
accessory ingredients. In general, the pharmaceutical compositions
are prepared by uniformly and intimately bringing the active
ingredient into association with a liquid carrier or a finely
divided solid carrier or both, and then, if necessary, shaping the
product into the desired formulation. In the pharmaceutical
composition the active object compound is included in an amount
sufficient to produce the desired effect upon the process or
condition of diseases. The pharmaceutical compositions containing
the active ingredient may be in a form suitable for oral use, for
example, as tablets, troches, lozenges, aqueous or oily
suspensions, dispersible powders or granules, emulsions, hard or
soft capsules, or syrups or elixirs.
[0307] Compositions intended for oral use may be prepared according
to any method known to the art for the manufacture of
pharmaceutical compositions and such compositions may contain one
or more agents selected from the group consisting of sweetening
agents, flavoring agents, coloring agents and preserving agents in
order to provide pharmaceutically elegant and palatable
preparations. Tablets contain the active ingredient in admixture
with non-toxic pharmaceutically acceptable excipients which are
suitable for the manufacture of tablets. These excipients may be
for example, inert diluents, such as calcium carbonate, sodium
carbonate, lactose, calcium phosphate or sodium phosphate;
granulating and disintegrating agents, for example, corn starch, or
alginic acid; binding agents, for example starch, gelatin or
acacia, and lubricating agents, for example magnesium stearate,
stearic acid or talc. The tablets may be uncoated or they may be
coated by known techniques to delay disintegration and absorption
in the gastrointestinal tract and thereby provide a sustained
action over a longer period. For example, a time delay material
such as glyceryl monostearate or glyceryl distearate may be
employed. They may also be coated to form osmotic therapeutic
tablets for control release.
[0308] Formulations for oral use may also be presented as hard
gelatin capsules wherein the active ingredient is mixed with an
inert solid diluent, for example, calcium carbonate, calcium
phosphate or kaolin, or as soft gelatin capsules wherein the active
ingredient is mixed with water or an oil medium, for example peanut
oil, liquid paraffin, or olive oil.
[0309] Aqueous suspensions contain the active materials in
admixture with excipients suitable for the manufacture of aqueous
suspensions. Such excipients are suspending agents, for example
sodium carboxymethylcellulose, methylcellulose,
hydroxy-propylmethylcellulose, sodium alginate,
polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or
wetting agents may be a naturally-occurring phosphatide, for
example lecithin, or condensation products of an alkylene oxide
with fatty acids, for example polyoxyethylene stearate, or
condensation products of ethylene oxide with long chain aliphatic
alcohols, for example heptadecaethyleneoxycetanol, or condensation
products of ethylene oxide with partial esters derived from fatty
acids and a hexitol such as polyoxyethylene sorbitol monooleate, or
condensation products of ethylene oxide with partial esters derived
from fatty acids and hexitol anhydrides, for example polyethylene
sorbitan monooleate. Also useful as a solubilizer is polyethylene
glycol, for example. The aqueous suspensions may also contain one
or more preservatives, for example ethyl, or n-propyl,
p-hydroxybenzoate, one or more coloring agents, one or more
flavoring agents, and one or more sweetening agents, such as
sucrose or saccharin.
[0310] Oily suspensions may be formulated by suspending the active
ingredient in a vegetable oil, for example arachis oil, olive oil,
sesame oil or coconut oil, or in a mineral oil such as liquid
paraffin. The oily suspensions may contain a thickening agent, for
example beeswax, hard paraffin or cetyl alcohol. Sweetening agents
such as those set forth above, and flavoring agents may be added to
provide a palatable oral preparation. These compositions may be
preserved by the addition of an anti-oxidant such as ascorbic
acid.
[0311] Dispersible powders and granules suitable for preparation of
an aqueous suspension by the addition of water provide the active
ingredient in admixture with a dispersing or wetting agent,
suspending agent and one or more preservatives. Suitable dispersing
or wetting agents and suspending agents are exemplified by those
already mentioned above. Additional excipients, for example
sweetening, flavoring and coloring agents, may also be present.
[0312] Syrups and elixirs may be formulated with sweetening agents,
for example glycerol, propylene glycol, sorbitol or sucrose. Such
formulations may also contain a demulcent, a preservative and
flavoring and coloring agents.
[0313] The pharmaceutical compositions may be in the form of a
sterile injectable aqueous or oleagenous suspension. This
suspension may be formulated according to the known art using those
suitable dispersing or wetting agents and suspending agents which
have been mentioned above. The sterile injectable preparation may
also be a sterile injectable solution or suspension in a
parenterally-acceptable diluent or solvent or coslvent or
complexing agent or dispersing agent or excipient or combination
thereof, for example 1,3-butane diol, polyethylene glycols,
polypropylene glycols, ethanol or other alcohols, povidones,
Tweens, sodium dodecyle sulfate, sodium deoxycholate,
dimethylacetamide, polysorbates, poloxamers, cyclodextrins, lipids,
and excipients such as inorganic salts (e.g., sodium chloride),
buffering agents (e.g., sodium citrate, sodium phosphate), and
sugars (e.g., saccharose and dextrose). Among the acceptable
vehicles and solvents that may be employed are water, dextrose
solutions, Ringer's solutions and isotonic sodium chloride
solution. In addition, sterile, fixed oils are conventionally
employed as a solvent or suspending medium. For this purpose any
bland fixed oil may be employed including synthetic mono- or
diglycerides. In addition, fatty acids such as oleic acid find use
in the preparation of injectables.
[0314] Depending on the condition being treated, these
pharmaceutical compositions may be formulated and administered
systemically or locally. Techniques for formulation and
administration may be found in the latest edition of "Remington's
Pharmaceutical Sciences" (Mack Publishing Co, Easton Pa.). Suitable
routes may, for example, include oral or transmucosal
administration; as well as parenteral delivery, including
intramuscular, subcutaneous, intramedullary, intrathecal,
intraventricular, intravenous, intraperitoneal, or intranasal
administration. For injection, the pharmaceutical compositions of
the invention may be formulated in aqueous solutions, preferably in
physiologically compatible buffers such as Hanks' solution,
Ringer's solution, or physiologically buffered saline. For tissue
or cellular administration, penetrants appropriate to the
particular barrier to be permeated are used in the formulation.
Such penetrants are generally known in the art. Pharmaceutical
formulations for parenteral administration include aqueous
solutions of the active compounds in water-soluble form.
Additionally, suspensions of the active compounds may be prepared
as appropriate oily injection suspensions. Suitable lipophilic
solvents or vehicles include fatty oils such as sesame oil, or
synthetic fatty acid esters, such as ethyl oleate or triglycerides,
or liposomes. Aqueous injection suspensions may contain substances
that increase the viscosity of the suspension, such as sodium
carboxymethyl cellulose, sorbitol, or dextran. Optionally, the
suspension may also contain suitable stabilizers or agents that
increase the solubility of the compounds to allow for the
preparation of highly concentrated solutions.
[0315] The compounds of the present invention may also be
administered in the form of suppositories for rectal administration
of the drug. These compositions can be prepared by mixing the drug
with a suitable non-irritating excipient which is solid at ordinary
temperatures but liquid at the rectal temperature and will
therefore melt in the rectum to release the drug. Such materials
are cocoa butter and polyethylene glycols.
[0316] For topical use, creams, ointments, jellies, solutions or
suspensions, etc., containing the compounds of the present
invention are employed. (For purposes of this application, topical
application shall include mouthwashes and gargles).
[0317] In one aspect, the invention compounds are administered in
combination with an antiinflammatory, antihistamines,
chemotherapeutic agent, immunomodulator, therapeutic antibody or a
protein kinase inhibitor, e.g., a tyrosine kinase inhibitor, to a
subject in need of such treatment. While not wanting to be
limiting, chemotherapeutic agents include antimetabolites, such as
methotrexate, DNA cross-linking agents, such as
cisplatin/carboplatin; alkylating agents, such as canbusil;
topoisomerase I inhibitors such as dactinomycin; microtubule
inhibitors such as taxol (paclitaxol), and the like. Other
chemotherapeutic agents include, for example, a vinca alkaloid,
mitomycin-type antibiotic, bleomycin-type antibiotic, antifolate,
colchicine, demecolcine, etoposide, taxane, anthracycline
antibiotic, doxorubicin, daunorubicin, caminomycin, epirubicin,
idarubicin, mitoxanthrone, 4-demethoxy-daunomycin,
11-deoxydaunorubicin, 13-deoxydaunorubicin, adriamycin-14-benzoate,
adriamycin-14-octanoate, adriamycin-14-naphthaleneacetate,
amsacrine, carmustine, cyclophosphamide, cytarabine, etoposide,
lovastatin, melphalan, topetecan, oxalaplatin, chlorambucil,
methotrexate, lomustine, thioguanine, asparaginase, vinblastine,
vindesine, tamoxifen, or mechlorethamine. While not wanting to be
limiting, therapeutic antibodies include antibodies directed
against the HER2 protein, such as trastuzumab; antibodies directed
against growth factors or growth factor receptors, such as
bevacizumab, which targets vascular endothelial growth factor, and
OSI-774, which targets epidermal growth factor; antibodies
targeting integrin receptors, such as Vitaxin (also known as
MEDI-522), and the like. Classes of anticancer agents suitable for
use in compositions and methods of the present invention include,
but are not limited to: 1) alkaloids, including, microtubule
inhibitors (e.g., Vincristine, Vinblastine, and Vindesine, etc.),
microtubule stabilizers (e.g., Paclitaxel [Taxol], and Docetaxel,
Taxotere, etc.), and chromatin function inhibitors, including,
topoisomerase inhibitors, such as, epipodophyllotoxins (e.g.,
Etoposide [VP-16], and Teniposide [VM-26], etc.), and agents that
target topoisomerase I (e.g., Camptothecin and Isirinotecan
[CPT-11], etc.); 2) covalent DNA-binding agents [alkylating
agents], including, nitrogen mustards (e.g., Mechlorethamine,
Chlorambucil, Cyclophosphamide, Ifosphamide, and Busulfan
[Myleran], etc.), nitrosoureas (e.g., Carmustine, Lomustine, and
Semustine, etc.), and other alkylating agents (e.g., Dacarbazine,
Hydroxymethylmelamine, Thiotepa, and Mitocycin, etc.); 3)
noncovalent DNA-binding agents [antitumor antibiotics], including,
nucleic acid inhibitors (e.g., Dactinomycin [Actinomycin D], etc.),
anthracyclines (e.g., Daunorubicin [Daunomycin, and Cerubidine],
Doxorubicin [Adriamycin], and Idarubicin [Idamycin], etc.),
anthracenediones (e.g., anthracycline analogues, such as,
[Mitoxantrone], etc.), bleomycins (Blenoxane), etc., and plicamycin
(Mithramycin), etc.; 4) antimetabolites, including, antifolates
(e.g., Methotrexate, Folex, and Mexate, etc.), purine
antimetabolites (e.g., 6-Mercaptopurine [6-MP, Purinethol],
6-Thioguanine [6-TG], Azathioprine, Acyclovir, Ganciclovir,
Chlorodeoxyadenosine, 2-Chlorodeoxyadenosine [CdA], and
2'-Deoxycoformycin [Pentostatin], etc.), pyrimidine antagonists
(e.g., fluoropyrimidines [e.g., 5-fluorouracil (Adrucil),
5-fluorodeoxyuridine (FdUrd) (Floxuridine)] etc.), and cytosine
arabinosides (e.g., Cytosar [ara-C] and Fludarabine, etc.); 5)
enzymes, including, L-asparaginase, and hydroxyurea, etc.; 6)
hormones, including, glucocorticoids, such as, antiestrogens (e.g.,
Tamoxifen, etc.), nonsteroidal antiandrogens (e.g., Flutamide,
etc.), and aromatase inhibitors (e.g., anastrozole [Arimidex],
etc.); 7) platinum compounds (e.g., Cisplatin and Carboplatin,
etc.); 8) monoclonal antibodies conjugated with anticancer drugs,
toxins, and/or radionuclides, etc.; 9) biological response
modifiers (e.g., interferons [e.g., IFN-.alpha., etc.] and
interleukins [e.g., IL-2, etc.], etc.); 10) adoptive immunotherapy;
11) hematopoietic growth factors; 12) agents that induce tumor cell
differentiation (e.g., all-trans-retinoic acid, etc.); 13) gene
therapy techniques; 14) antisense therapy techniques; 15) tumor
vaccines; 16) therapies directed against tumor metastases (e.g.,
Batimistat, etc.); and 17) inhibitors of angiogenesis.
[0318] The pharmaceutical composition and method of the present
invention may further comprise other therapeutically active
compounds as noted herein which are usually applied in the
treatment of the above mentioned pathological conditions. Examples
of other therapeutic agents include the following: cyclosporine
(e.g., cyclosporin A), CTLA4-Ig, antibodies such as ICAM-3,
anti-IL-2 receptor (Anti-Tac), anti-CD45RB, anti-CD2, anti-CD3
(OKT-3), anti-CD4, anti-CD80, anti-CD86, agents blocking the
interaction between CD40 and gp39, such as antibodies specific for
CD40 and/or gp39 (i.e., CD154), fusion proteins constructed from
CD40 and gp39 (CD40Ig and CD8gp39), inhibitors, such as nuclear
translocation inhibitors, of NF-kappa B function, such as
deoxyspergualin (DSG), cholesterol biosynthesis inhibitors such as
HMG CoA reductase inhibitors (lovastatin and simvastatin),
non-steroidal antiinflammatory drugs (NSAIDs) such as ibuprofen and
cyclooxygenase inhibitors such as rofecoxib, steroids such as
prednisone or dexamethasone, gold compounds, antiproliferative
agents such as methotrexate, FK506 (tacrolimus, Prograf),
mycophenolate mofetil, cytotoxic drugs such as azathioprine and
cyclophosphamide, TNF-a inhibitors such as tenidap, anti-TNF
antibodies or soluble TNF receptor, and rapamycin (sirolimus or
Rapamune) or derivatives thereof.
[0319] Other agents that may be administered in combination with
invention compounds include protein therapeutic agents such as
cytokines, immunomodulatory agents and antibodies. As used herein
the term "cytokine" encompasses chemokines, interleukins,
lymphokines, monokines, colony stimulating factors, and receptor
associated proteins, and functional fragments thereof. As used
herein, the term "functional fragment" refers to a polypeptide or
peptide which possesses biological function or activity that is
identified through a defined functional assay.
[0320] The cytokines include endothelial monocyte activating
polypeptide II (EMAP-II), granulocyte-macrophage-CSF (GM-CSF),
granulocyte-CSF (G-CSF), macrophage-CSF (M-CSF), IL-1, IL-2, IL-3,
IL-4, IL-5, IL-6, IL-12, and IL-13, interferons, and the like and
which is associated with a particular biologic, morphologic, or
phenotypic alteration in a cell or cell mechanism.
[0321] The term antibody as used in this invention is meant to
include intact molecules of polyclonal or monoclonal antibodies, as
well as fragments thereof, such as Fab and F(ab').sub.2, Fv and SCA
fragments which are capable of binding an epitopic determinant.
[0322] When other therapeutic agents are employed in combination
with the compounds of the present invention they may be used for
example in amounts as noted in the Physician Desk Reference (PDR)
or as otherwise determined by one of ordinary skill in the art.
[0323] In the treatment or prevention of conditions which involve
compromised vasculostasis an appropriate dosage level will
generally be about 0.01 to 500 mg per kg patient body weight per
day which can be administered in single or multiple doses.
Preferably, the dosage level will be about 0.01 to about 250 mg/kg
per day; more preferably about 0.5 to about 100 mg/kg per day. A
suitable dosage level may be about 0.01 to 250 mg/kg per day, about
0.05 to 100 mg/kg per day, or about 0.1 to 50 mg/kg per day or 1.0
mg/kg per day. Within this range the dosage may be 0.05 to 0.5, 0.5
to 5 or 5 to 50 mg/kg per day for example. The Examples section
shows that one of the exemplary compounds was preferred at 0.1
mg/kg/day while another was effective at about 1.0 mg/kg/day. For
oral administration, the compositions are preferably provided in
the form of tablets containing 1.0 to 1000 milligrams of the active
ingredient, particularly 1.0, 5.0, 10.0, 15.0. 20.0, 25.0, 50.0,
75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0,
750.0, 800.0, 900.0, and 1000.0 milligrams of the active ingredient
for the symptomatic adjustment of the dosage to the patient to be
treated. The compounds may be administered on a regimen of 1 to 4
times per day, preferably once or twice per day. There may be a
period of no administration followed by another regimen of
administration. Preferably, administration of the compound is
closely associated with the schedule of IL-2 administration. For
example, administration can be prior to, simultaneously with or
immediately following IL-2 administration
[0324] It will be understood, however, that the specific dose level
and frequency of dosage for any particular patient may be varied
and will depend upon a variety of factors including the activity of
the specific compound employed, the metabolic stability and length
of action of that compound, the age, body weight, general health,
sex, diet, mode and time of administration, rate of excretion, drug
combination, the severity of the particular condition, and the host
undergoing therapy.
[0325] Another embodiment described herein is based on the
discovery that a compound that is a vasculostatic agent alone or in
combination with an effective amount of therapeutic antibody (or
therapeutic fragment thereof), chemotherapeutic or immunotoxic
agent, is an effective therapeutic regimen for treatment of tumors,
for example. While doxorubicin, docetaxel, or taxol are described
in the present application as illustrative examples of
chemotherapeutic agents, it should be understood that the invention
includes combination therapy including a compound of the invention,
including but not limited to vasculostatic agents, such as
tyrosine, serine or threonine kinase inhibitors, for example,
Src-family inhibitors, and any chemotherapeutic agent or
therapeutic antibody.
[0326] Such vasculostatic agents, in combination with
chemotherapeutic agents or therapeutic antibodies are effective in
blocking vascular permeability and/or vascular leakage and/or
angiogenesis. In one embodiment, the invention provides a
composition containing a therapeutically effective amount of a
chemotherapeutic agent and a vasculostatic agent in a
pharmaceutically acceptable carrier.
[0327] In one embodiment, the invention provides a method for
reducing the tumor burden in a subject, comprising administering to
a subject in need thereof an effective amount of chemotherapeutic
agent in combination with a compound that is a vasculostatic agent.
In an illustrative example, the method includes use of at least one
of the invention compounds e.g., as set forth in Structures I, II,
III, IIIa, IV, V, VI or VII or any combination thereof, with the
chemotherapeutic agent. In one aspect, the compound is set forth in
FIG. 1. It should be understood that the tumor burden in a subject
can be reduced prior to treatment with a compound of the invention
through surgical resection, chemotherapy, radiation treatment or
other methods known to those of skill in the art.
[0328] The compounds according to this invention may contain one or
more asymmetric carbon atoms and thus occur as racemates and
racemic mixtures, single enantiomers, diastereomeric mixtures and
individual diastereomers. The term "stereoisomer" refers to a
chemical compounds which differ from each other only in the way
that the different groups in the molecules are oriented in space.
Stereoisomers have the same molecular weight, chemical composition,
and constitution as another, but with the atoms grouped
differently. That is, certain identical chemical moieties are at
different orientations in space and, therefore, when pure, have the
ability to rotate the plane of polarized light. However, some pure
stereoisomers may have an optical rotation that is so slight that
it is undetectable with present instrumentation. All such isomeric
forms of these compounds are included in the present invention.
[0329] Each stereogenic carbon may be of R or S configuration.
Although the specific compounds exemplified in this application may
be depicted in a particular configuration, compounds having either
the opposite stereochemistry at any given chiral center or mixtures
thereof are also envisioned. When chiral centers are found in the
derivatives of this invention, it is to be understood that this
invention encompasses all possible stereoisomers. The terms
"optically pure compound" or "optically pure isomer" refers to a
single stereoisomer of a chiral compound regardless of the
configuration of the compound.
[0330] Several illustrative compounds employed in the methods of
the present invention are inhibitors of kinases and therefore are
useful in treating a wide variety of disorders resulting from
aberrant kinase activity. Examples of kinases include Src-family
tyrosine kinases and their associated disorders, which result from
aberrant tyrosine kinase activity, and/or which are alleviated by
the inhibition of one or more of the enzymes within the Src family.
For example, Src inhibitors are of value in the treatment of
cancer, as Src inhibition blocks tumor cell migration and survival.
Many compounds of the invention are also broad spectrum kinase
inhibitors and inhibit other kinases in addition to Src-family
tyrosine kinases or non-Src family kinases.
[0331] Cancers that may be treated by compounds of the invention
alone or as a combination therapy of the invention include but are
not limited to a carcinoma or a sarcoma, including one or more
specific types of cancer, e.g., an alimentary/gastrointestinal
tract cancer, a liver cancer, a skin cancer, a breast cancer, an
ovarian cancer, a prostate cancer, a lymphoma, a leukemia, a kidney
cancer, a lung cancer, a muscle cancer, a bone cancer, bladder
cancer or a brain cancer.
[0332] The present invention also provides articles of manufacture
comprising packaging material and a pharmaceutical composition
contained within said packaging material, wherein said packaging
material comprises a label which indicates that said pharmaceutical
composition can be used for treatment of disorders and wherein said
pharmaceutical composition comprises a compound according to the
present invention. Thus, in one aspect, the invention provides a
pharmaceutical composition including both a chemotherapeutic agent,
immunotoxin or therapeutic antibody and a compound of the invention
(e.g, as shown in FIG. 1), wherein the compound is present in a
concentration effective to reduce tumor burden, for example. In one
aspect, the invention provides a pharmaceutical composition
including a compound of the invention, wherein the compound is
present in a concentration effective to reduce vascular
permeability, for example. The concentration can be determined by
one of skill in the art according to standard treatment regimen or
as determined by an in vivo animal assay, for example.
[0333] Pharmaceutical compositions employed as a component of
invention articles of manufacture can be used in the form of a
solid, a solution, an emulsion, a dispersion, a micelle, a
liposome, and the like, wherein the resulting composition contains
one or more of the compounds described above as an active
ingredient, in admixture with an organic or inorganic carrier or
excipient suitable for enteral or parenteral applications.
Compounds employed for use as a component of invention articles of
manufacture may be combined, for example, with the usual non-toxic,
pharmaceutically acceptable carriers for tablets, pellets,
capsules, suppositories, solutions, emulsions, suspensions, and any
other form suitable for use. The carriers which can be used include
glucose, lactose, gum acacia, gelatin, mannitol, starch paste,
magnesium trisilicate, talc, corn starch, keratin, colloidal
silica, potato starch, urea, medium chain length triglycerides,
dextrans, and other carriers suitable for use in manufacturing
preparations, in solid, semisolid, or liquid form. In addition
auxiliary, stabilizing, thickening and coloring agents and perfumes
may be used.
[0334] The present invention also provides pharmaceutical
compositions including at least one invention compound in an amount
effective for treating a tumor, or cancer, alone or in combination
with a chemotherapeutic agent, immunotoxin, immunomodulator or
therapeutic antibody and a pharmaceutically acceptable vehicle or
diluent. Similarly, the present invention provides pharmaceutical
compositions including at least one invention compound capable of
treating a disorder associated with vasculostasis in an amount
effective therefore. The compositions of the present invention may
contain other therapeutic agents as described herein and may be
formulated, for example, by employing conventional solid or liquid
vehicles or diluents, as well as pharmaceutical additives of a type
appropriate to the mode of desired administration (for example,
excipients, binders, preservatives, stabilizers, flavors, etc.)
according to techniques such as those well known in the art of
pharmaceutical formulation.
[0335] The terms "administration of" and or "administering a"
compound should be understood to mean providing a compound of the
invention or pharmaceutical composition to the subject in need of
treatment. For example, administration of the vasculostatic agent
can be prior to, simultaneously with, or after administration of an
invention compound or other agent. In the Examples provided herein,
typically the compounds of the invention are co-administered at the
same time as a chemotherapeutic agent.
[0336] While not wanting to be limiting, chemotherapeutic agents
include antimetabolites, such as methotrexate, DNA cross-linking
agents, such as cisplatin/carboplatin; alkylating agents, such as
canbusil; topoisomerase I inhibitors such as dactinomycin;
microtubule inhibitors such as taxol (paclitaxol), and the like.
Other chemotherapeutic agents include, for example, a vinca
alkaloid, mitomycin-type antibiotic, bleomycin-type antibiotic,
antifolate, amsacrine, carmustine, cyclophosphamide, cytarabine,
etoposide, lovastatin, melphalan, topetecan, oxalaplatin,
chlorambucil, methotrexate, lomustine, thioguanine, asparaginase,
vinblastine, vindesine, tamoxifen, mechlorethamine, colchicine,
demecolcine, etoposide, taxane, anthracycline antibiotic,
doxorubicin, daunorubicin, caminomycin, epirubicin, idarubicin,
mitoxanthrone, 4-demethoxy-daunomycin, 11-deoxydaunorubicin,
13-deoxydaunorubicin, adriamycin-14-benzoate,
adriamycin-14-octanoate or adriamycin-14-naphthaleneacetate.
[0337] Compounds, their prodrugs, or metabolites employed in the
methods of the present invention are vasculostatic agents such as
inhibitors of vascular permeability and/or vascular leakage and/or
angiogenesis. In addition, several illustrative compounds employed
in the methods of the present invention are inhibitors of kinases
and therefore are useful in treating a wide variety of disorders
resulting from aberrant kinase activity. Kinase-associated
disorders are those disorders which result from aberrant kinase
activity, and/or which are alleviated by the inhibition of one or
more of the kinases.
[0338] It will be understood, however, that the specific dose level
and frequency of dosage for any particular patient may be varied
and will depend upon a variety of factors including the activity of
the specific compound employed, the metabolic stability and length
of action of that compound, the age, body weight, general health,
sex, diet, mode and time of administration, rate of excretion, drug
combination, the severity of the particular condition, and the host
undergoing therapy.
[0339] The Examples set out below include representative examples
of aspects of the present invention. The Examples are not meant to
limit the scope of the invention but rather serve examplery
purposes. In addition, various aspects of the invention can be
summarized by the following description. However, this description
is not meant to limit the scope of the invention but rather to
highlight various aspects of the invention. One having ordinary
skill in the art can readily appreciate additional aspects and
embodiments of the invention.
Example 1
Syntheses of Vasculostatic Agents
Experimentals
General Analytical Methods
[0340] All solvents are used without further purification.
Reactions are usually run without an inert gas atmosphere unless
specified otherwise. All .sup.1H NMR are run on a 500 MHz Bruker
NMR. Chemical shifts are reported in delta (.delta.) units, parts
per million (ppm) downfield from tetramethylsilane. Coupling
constants are reported in hertz (Hz). A Water LC/MS system is used
in identity and purity analysis. This system includes a 2795
separation module, a 996 photodidode array detector and a ZQ2000
mass spectrometer. A Zorbax SB column (150.times.4.6 mm 3.5.mu.,
Agilent Technologies) is used for the LC. Column temperature is
40.degree. C. Compounds are separated using gradient elution with
mobile phases of water (0.05% TFA (A)) and acetonitrile (0.05% TFA
(B)). Flow rate is 1 mL/min. The gradient program used in
separation is 0-15 min: 5-60% B; 15-15.5 min: 60-100% B; 15.5-17
min: 100% B.
[0341] The following gradient programs were used based on the
hydrophobicity of the analyzed sample: (1) 0-15 min: 30-70% B;
15-15.5 min: 70-90% B; 15.5-17 min: 90% B for the compounds:
4-Hydroxy-N-(2-(1H-indol-2-yl)-phenyl)-benzamide;
3,4-Dihydroxy-N-(2-(1H-indol-2-yl)-phenyl)-benzamide;
N-(2-(1H-Indol-2-yl)-phenyl)-2-phenyl-acetamide;
2-(3,4-Dihydroxy-phenyl)-N-(2-(1H-indol-2-yl)-phenyl)-acetamide;
N-(2-(1H-Indol-2-yl)-phenyl)-3-phenyl-propionamide;
3-(4-Hydroxy-phenyl)-N-(2-(1H-indol-2-yl)-phenyl)-propionamide;
N-(2-(1H-Indol-2-yl)-phenyl)-3-(2-methoxy-phenyl)-propionamide;
3-(3,4-Dihydroxy-phenyl)-N-(2-(1H-indol-2-yl)-phenyl)-propionamide;
(2) 0-15 min: 30-50% B; 15-15.5 min: 50-90% B; 15.5-17 min: 90% B
for compound
N-(2-(2,3-Dihydro-1H-indol-2-yl)-phenyl)-2-hydroxy-benzamide. (3)
0-15 min: 20-40% B; 15-15.5 min: 40-90% B; 15.5-17 min: 90% B for
compound
4-(4-amino-6-(3,4-dihydroxyphenyl)pteridin-7-yl)benzene-1,2-diol- .
(4) 0-15 min: 5-60% B; 15-15.5 min: 60-90% B; 15.5-17 min: 90% B
for compound
2-(4-Hydroxy-phenyl)-N-(2-(1H-indol-2-yl)-phenyl)-acetamide. (5)
0-15 min: 40-100% B; 15.5-17 min: 100% B for compounds
N-(2-(1H-Indol-2-yl)-phenyl)-2-(2-methoxy-phenyl)-acetamide and
2-Benzo(1,3)dioxol-5-yl-N-(2-(1H-indol-2-yl)-phenyl)-acetamide.
[0342] The mass spectrometer is equipped with an electrospray
probe. Source temperature is 120.degree. C. All of the compounds
are identified using the positive mode with mass scan range from
100 to 800.
General Procedure for Indoles
[0343] 2-(2-Aminophenyl) indole and the starting material acid (2
equiv) were dissolved in acetonitrile. To the solution were added 2
equiv of EDC (dimethylaminopropyl ethylcarbodiimide hydrochloride)
as powder. The mixture was stirred at either room temperature
(23.degree. C.) or at slightly elevated temperature (50.degree. C.)
for 3 to 16 hours.
[0344] The solvent was removed and the residue dissolved in
methanol:ethylacetate (5-10%). The solution was extracted with 1 M
HCl as well as saturated sodium bicarbonate solution. The aqueous
phases were re-extracted with EtOAc, respectively. The combined
organic phases were dried over magnesium sulfate. The product was
purified by column chromatography (silica, typically using
EtOAc-hexanes as mobile phase) and/or crystallization from
different solvents including methanol and acetonitrile.
2-(4-Hydroxy-phenyl)-N-[2-(1H-indol-2-yl)-phenyl]-acetamide
##STR00066##
[0346] 1 g (4.8 mmol) 2-(2-Aminophenyl) indole was dissolved in 200
ml acetonitrile. 1.46 g (9.6 mmol, 2 eq) of 4-hydroxyphenylacetic
acid were dissolved in 50 ml acetonitrile and added to the
solution. To the mixture were added 1.84 g (9.6 mmol, 2 eq) of EDC
(dimethylaminopropyl ethylcarbodiimide hydrochloride). The reaction
mixture was stirred at 23.degree. C. for 16 hours. The solvent was
removed and the residue was dissolved in 100 ml
ethylacetate:methanol (10:1). It was extracted twice with 100 ml of
aqueous 1M HCl as well as 100 ml of aqueous, saturated sodium
bicarbonate solution. The aqueous phases were re-extracted with
EtOAc, respectively. The combined organic phases were dried over
magnesium sulfate. The crude product was chromatographed on silica
using a ethylacetate/hexane gradient (10%-50%) to obtain 1.23 g of
the amide as a pink colored powder in an overall yield of 75%. 100%
Purity by LC/MS (230 DAD) Mass-spec [M+H.sup.+]=343.9 .sup.1H NMR
(MeOH-d4): 3.60 s (2H), 6.10 s (1H), 6.70 d, 8 Hz (2H), 7.03 t, 8
Hz (1H), 7.09-7.13 m (3H), 7.25 t, 7 Hz (1H), 7.34 m (2H), 7.49 d,
8 Hz (1H), 7.53 d, 8 Hz (1H), 7.95 d, 8 Hz (1H).
4-Hydroxy-N-(2-(1H-indol-2-yl)-phenyl)-benzamide
##STR00067##
[0348] Prepared from 2-(2-aminophenyl) indole and 4-hydroxybenzoic
acid in 35% overall yield following procedure 1. The product was
chromatographed on silica and crystallized from acetonitrile. 95.6%
Purity by LC/MS (230 DAD) Mass-spec (M+.sup.+)=329.8 .sup.1H NMR
(MeOH-d4): 6.65 s (1H), 6.83 m (2H), 7.01 t, 7 Hz (1H), 7.12 td,
7.1 Hz (1H), 7.34 td, 7, 1 Hz (1H), 7.39-7.43 m (2H), 7.51 d, 7 Hz
(1H), 7.66 dd, 8.1 Hz (1H), 7.76 m (2H), 7.91 dd, 8.1 Hz (1H).
3,4-Dihydroxy-N-(2-(1H-indol-2-yl)-phenyl)-benzamide
##STR00068##
[0350] Prepared from 2-(2-aminophenyl) indole and
3,4-dihydroxybenzoic acid in 54% yield following procedure 1. The
product was chromatographed on silica. 100% Purity by LC/MS (230
DAD), Mass-spec (M+H.sup.+)=345.83, .sup.1H NMR (MeOH-d4): 6.645 s
(1H), 6.80 d, 8 Hz (1H), 7.02 t, 8 Hz (1H), 7.12 td, 8, 1 Hz (1H),
7.23 dd, 8, 1 Hz (1H), 7.33-7.36 m (2H), 7.39-7.42 m (2H), 7.52 d,
7 Hz (1H), 7.65 dd, 8, 1 Hz (1H), 7.94 d, 8 Hz (1H).
2-Hydroxy-N-(2-(1H-indol-2-yl)-phenyl)-benzamide
##STR00069##
[0352] Prepared from 2-(2-aminophenyl) indole and salicylic acid in
46% yield following procedure 1. The compound was chromatographed
on silica using an ethylacetate/hexane gradient. % Purity by LC/MS
(230 DAD), Mass-spec (M+H.sup.+)=329, .sup.1H NMR (MeOH-d4): 6.66 s
(1H), 6.86 dd,
N-[2-(1H-Indol-2-yl)-phenyl]-2-phenyl-acetamide
##STR00070##
[0354] Prepared from 2-(2-aminophenyl) indole and phenylacetic acid
in 62% yield following procedure 1. The product was crystallized
from methanol. 100% Purity by LC/MS (230 DAD), Mass-spec
[M+H.sup.+]=327, .sup.1H NMR (MeOH-d4): 3.69 s (2H), 6.21 s (1H),
7.03 t, 7 Hz (1H), 7.12 t, 8 Hz (1H), 7.21-7.28 m (6H), 7.33-7.36 m
(2H), 7.46 d, 8 Hz (1H), 7.54 dd, 7.1 Hz (1H), 7.89 d, 8 Hz
(1H).
N-[2-(1H-Indol-2-yl)-phenyl]-2-(2-methoxy-phenyl)-acetamide
##STR00071##
[0356] Prepared from 2-(2-aminophenyl) indole and
2-methoxyphenylacetic acid in 53% yield following procedure 1. The
product was crystallized from acetonitrile. 100% Purity by LC/MS
(230 DAD), Mass-spec [M+H.sup.+]=357, .sup.1H NMR (MeOH-d4): 3.45 s
(3H, OMe), 3.67 s (2H), 6.17 s (1H), 6.75 d, 8 Hz (1H), 6.83 t, 8
Hz (1H), 7.06 t, 8 Hz (1H), 7.14 t, 8 Hz (1H), 7.17-7.21 m (3H),
7.23-7.36 m (2H), 7.49 t, 8 Hz (2H), 8.13 d, 8 Hz (1H).
2-(2-Hydroxy-phenyl)-N-[2-(1H-indol-2-yl)-phenyl]-acetamide
##STR00072##
[0358] The product was prepared from
N-[2-(1H-Indol-2-yl)-phenyl]-2-(2-methoxy-phenyl)-acetamide.
Cleavage of the methylether was accomplished using 1.8 eq of
BBr.sub.3 (1M solution in dichloromethane) at -78.degree. C. to
room temperature (23.degree. C.) and subsequent hydrolysis (32%
yield). 96% Purity by HPLC (ELSD), Mass-spec [M+H.sup.+]=343,
.sup.1H NMR (MeOH-d4): 3.69 s (2H), 6.25 s (1H), 6.71-6.74 m (2H),
7.01-7.07 m (2H), 7.10-7.13 m (2H), 7.22 t, 7 Hz (1H), 7.31-7.36 m
(2H), 7.48 d, 8 Hz (1H), 7.52 dd, 8, 1 Hz (1H), 8.08 d, 8 Hz
(1H).
2-(3,4-Dihydroxy-phenyl)-N-[2-(1H-indol-2-yl)-phenyl]-acetamide
##STR00073##
[0360] Prepared from 2-(2-aminophenyl) indole and
3,4-dihydroxyphenylacetic acid in 17% yield. The product was
chromatographed on silica. 100% Purity by LC/MS (230 DAD),
Mass-spec [M+H.sup.+]=359, .sup.1H NMR (MeOH-d4): 3.56 s (2H), 6.10
s (1H), 6.59 dd, 8, 2 Hz (1H), 6.66 d, 8 Hz (1H), 6.78 d, 2 Hz
(1H), 7.03 t, 8 Hz (1H), 7.11 t, 8 Hz (1H), 7.25 t, 8 Hz (1H),
7.31-7.35 m (2H), 7.51 d, 7 Hz (1H), 7.55 dd, 8, 1 Hz (1H), 7.99 d,
8 Hz (1H).
2-Benzo[1,3]dioxol-5-yl-N-[2-(1H-indol-2-yl)-phenyl]-acetamide
##STR00074##
[0362] Prepared from 2-(2-aminophenyl) indole and
3,4-(methylenedioxy)phenylacetic acid in 55% yield. The product was
purified by crystallization from acetonitrile. 100% Purity by LC/MS
(230 DAD), Mass-spec [M+H.sup.+]=371, .sup.1H NMR (MeOH-d4): 3.61 s
(2H), 5.82 s (2H), 6.20 s (1H), 6.66 d, 8 Hz (1H), 6.74 dd, 8.1 Hz
(1H), 6.76 d, 1 Hz (1H), 7.03 t, 8 Hz (1H), 7.12 t, 8 Hz (1H), 7.25
t, 8 Hz (1H), 7.33-7.36 m (2H), 7.48 d, 8 Hz (1H), 7.52 d, 8 Hz
(1H), 7.99 d, 8 Hz (1H).
N-[2-(1H-Indol-2-yl)-phenyl]-3-phenyl-propionamide
##STR00075##
[0364] Prepared from 2-(2-aminophenyl) indole and hydrocinnamic
acid in 54% yield following procedure 1. The product was
crystallized from methanol. 99% Purity by LC/MS (230 DAD),
Mass-spec [M+H]=341, .sup.1H NMR (DMSO-d6): 2.65 t, 7.5 Hz (2H),
2.91 t, 7.5 Hz (2H), 6.50 s (1H), 7.00 t, 7 Hz (1H), 7.10 t, 7 Hz
(1H), 7.19-7.34 m (7H), 7.39 d, 8 Hz (1H), 7.51 d, 8 Hz (1H),
7.60-7.62 m (2H), 9.39 s (1H), 11.32 s (1H).
3-(4-Hydroxy-phenyl)-N-[2-(1H-indol-2-yl)-phenyl]-propionamide
##STR00076##
[0366] Prepared from 2-(2-aminophenyl) indole and
3-(4-hydroxyphenyl) propionic acid in 55% yield following procedure
1. The product was chromatographed on silica and crystallized from
acetonitrile. 100% Purity by LC/MS (230 DAD), Mass-spec
[M+H.sup.+]=357, .sup.1H NMR (MeOH-d4): 2.61 t, 7.4 Hz (1H), 2.89
t, 7.4 Hz (1H), 6.37 s (1H), 6.72 d, 8 Hz (2H), 7.00-7.06 m (3H),
7.11 t, 7 Hz (1H), 7.27-7.35 m (2H), 7.38 d, 8 Hz (1H), 7.54 d, 7
Hz (1H), 7.58 dd, 7.1 Hz (1H), 7.67 d, 8 Hz (1H).
N-[2-(1H-Indol-2-yl)-phenyl]-3-(2-methoxy-phenyl)-propionamide
##STR00077##
[0368] Prepared from 2-(2-aminophenyl) indole and
3-(2-methoxyphenyl) propionic acid in 62% yield following procedure
1. The product was crystallized from acetonitrile. 96% Purity by
LC/MS (TIC, DAD), Mass-spec [M+H.sup.+]=371, .sup.1H NMR (MeOH-d4):
2.62 t, 7.5 Hz (2H), 2.97 t, 7.5 Hz (2H), 3.74 s (3H, OMe), 6.40 s
(1H), 6.81 t, 7 Hz (1H), 6.88 d, 8 Hz (1H), 7.03 t, 8 Hz (1H),
7.10-7.14 m (2H), 7.17 t, 8 Hz (1H), 7.27 t, 7 Hz (1H), 7.33 td,
7.5, 1 Hz (1H), 7.40 d, 8 Hz (1H), 7.54 d, 8 Hz (1H), 7.57 dd, 7.1
Hz (1H), 7.76 d, 8 Hz (1H).
3-(3,4-Dihydroxy-phenyl)-N-[2-(1H-indol-2-yl)-phenyl]-propionamide
##STR00078##
[0370] Prepared from 2-(2-aminophenyl) indole and
3,4-dihydroxyhydrocinnamic acid in 19% yield following procedure 1.
The product was chromatographed on silica and crystallized from
acetonitrile. 100% Purity by LC/MS (230 DAD), Mass-spec
[M+H.sup.+]=373, NMR (MeOH-d4): 2.60 t, 7.4 Hz (2H), 2.85 t, 7.4 Hz
(2H), 6.38 s (1H), 6.55 dd, 8.2 Hz (1H), 6.69 m (2H), 7.02 t, 8 Hz
(1H), 7.11 t, 8 Hz (1H), 7.27-7.35 m (2H), 7.38 d, 8 Hz (1H), 7.56
d, 8 Hz (1H), 7.58 dd, 7.1 Hz (1H), 7.70 d, 8 Hz (1H).
2-(4-Hydroxy-phenoxy)-N-[2-(1H-indol-2-yl)-phenyl]-acetamide
##STR00079##
[0372] Prepared from 2-(2-aminophenyl) indole and
(4-hydroxyphenoxy)acetic acid in 30% yield following procedure 1.
The product was crystallized from methanol. 89% Purity by LC/MS
(230 DAD), Mass-spec [M+H.sup.+]=359, .sup.1H NMR (MeOH-d4): 4.52 s
(2H), 6.55 d, 9 Hz (2H), 6.58 s (1H), 6.61 d, 9 Hz (2H), 7.09 t, 8
Hz (1H), 7.18 t, 8 Hz (1H), 7.26 t, 8 Hz (1H), 7.37-7.43 m (2H),
7.56 t, 8 Hz (2H), 8.38 d, 8 Hz (1H).
2-Acetylamino-3-(4-hydroxy-phenyl)-N-[2-(1H-indol-2-yl)-phenyl]-propionami-
de
##STR00080##
[0374] Prepared from 2-(2-aminophenyl) indole and
N-acetyl-L-tyrosine in 69% yield following procedure 1. The product
was chromatographed on silica. 99% Purity by LC/MS (230 DAD),
Mass-spec [M+H.sup.+]=414, .sup.1H NMR (MeOH-d4): 1.79 s (3H,
COMe), 2.83 dd, 14.9 Hz (1H), 3.14 dd, 14.6 Hz (1H), 4.58 dd, 9.6
Hz (1H), 6.51 s (1H), 6.70 d, 8 Hz (2H), 7.02 t, 7.5 Hz (1H), 7.07
d, 8 Hz (2H), 7.12 td, 8.1 Hz (1H), 7.27 td, 8.1 Hz (1H), 7.33 td,
8.1 Hz (1H), 7.44 d, 8 Hz (1H), 7.56 d, 8 Hz (1H), 7.59 dd, 8.1 Hz
(1H), 7.83 d, 8 Hz (1H).
Procedure 2
N-[2-(1H-Indol-2-yl)-phenyl]-phthalamic acid
##STR00081##
[0376] 958 mg (4.6 mmol) 2-(2-Aminophenyl) indole and 675 mg (5.52
mmol, 1.2 eq) DMAP (dimethylamino pyridine) were dissolved in 35 ml
anhydrous dichloromethane. The mixture was stirred for 10 min. 954
mg (6.44 mmol, 1.4 eq) of phthalic anhydride in 3 ml anhydrous
dichloromethane were added and the mixture was stirred at
23.degree. C. for three hours. To the mixture were added 20 ml
dichloromethane. It was extracted with 50 ml aqueous 1 M HCl. The
aqueous phase was re-extracted with 30 ml dichloromethane. The
combined organic phases were dried over magnesium sulfate. The
crude product was chromatographed on silica using an
ethylacetate/hexane gradient (10%-90%) as mobile phase. The solvent
was removed and the product was re-crystallized from
ethylacetate:hexane (70:30) to obtain 654 mg of ivory colored
crystals in 40% overall yield.
[0377] 95% Purity by LC/MS (230 DAD), Mass-spec [M+H.sup.+]=357,
.sup.1H NMR (MeOH-d4): 6.75 s (1H), 6.99 t, 8 Hz (1H), 7.09 t, 7 Hz
(1H), 7.35-7.43 m (3H), 7.52-7.57 m (3H), 7.63 t, 8 Hz (1H), 7.71
d, 8 Hz (1H), 7.84 d, 8 Hz (1H), 8.06 d, 7 Hz (1H).
2-[2-(1H-Indol-2-yl)-phenylcarbamoyl]-nicotinic acid
##STR00082##
[0379] 104 mg (0.5 mmol) 2-(2-Aminophenyl) indole and 74 mg (0.6
mmol, 1.2 eq) DMAP (dimethylamino pyridine) were dissolved in 5 ml
anhydrous dichloromethane. The mixture was stirred for 10 min. 104
mg (0.7 mmol, 1.4 eq) of 2,3-pyridinedicarboxylic anhydride were
added and the mixture was stirred at 23.degree. C. for three
hours.
[0380] To the mixture were added 20 ml dichloromethane. It was
extracted with 20 ml saturated NaCl solution. The aqueous phase was
re-extracted with 20 ml dichloromethane. The combined organic
phases were dried over magnesium sulfate. The crude product was
chromatographed on silica and re-crystallized from acetonitrile.
100% Purity by HPLC (UV, 230 nm), Mass-spec [M+H.sup.+]=358,
.sup.1H NMR (MeOH-d4): 6.80 s (1H), 7.04 t, 7 Hz (1H), 7.14 t, 8 Hz
(1H), 7.31 t, 7 Hz (1H), 7.42 t (2H), 7.57 d, 8 Hz (1H), 7.61 dd,
8, 5 Hz (1H), 7.67 dd, 8, 1 Hz (1H), 8.13 dd, 8.1 Hz (1H), 8.30 d,
8 Hz (1H), 8.61 dd, 5, 1 Hz (1H).
3,4,5-trihydroxy-N-[2-(1H-indol-2-yl)-phenyl]-benzamide
##STR00083##
[0382] A 25-mL one-necked recovery flask equipped with a stirring
bar and a septum was charged with gallic acid (176 mg; 1.03 mmol;
1.00 equiv). A clear, colorless solution was formed on addition of
5 mL of dichloromethane. Solid EDC (197 mg; 1.03 mmol; 1.00 equiv)
and 2-(2-aminophenyl)indole (194 mg; 0.932 mmol; 0.904 equiv) were
added sequentially as solids. The reaction was worked up after 24 h
by extraction with 10 mL of NaHCO.sub.3 (satd aq). The organic
layer was dried (anhydrous sodium sulfate), filtered and
concentrated by rotary evaporation to yield a yellow oily paste.
The crude was purified using DCM-MeOH (19:1) to yield a light
yellow solid (230 mg; 68%).
[0383] Representative Syntheses of Compounds of Structure II
##STR00084##
[0384] A 100-mL, one-necked, round bottomed flask with a magnetic
stirring bar and a septum was charged with 2-(2-aminophenyl) indole
(210 mg; 1.01 mmol). The indole was dissolved in ca. 7 mL of
dichloromethane to give a very pale yellow solution. DMAP (143 mg;
1.17 mmol; 1.16 equiv) and phthalic anhydride (179 mg; 1.21 mmol;
1.20 equiv) were added sequentially each dissolving completely with
a resulting yellow solution. The solution was stirred at room
temperature, and the reaction was followed by TLC, and showed
complete conversion in ca. 30 min as indicated by the disappearance
of the 2-(2-aminophenyl) indole. The reaction mixture was poured
into a 125-mL separatory funnel and extracted with 15 mL HCl (aq,
ca. 1 M). The aqueous layer was washed with 2.times.5 mL
CH.sub.2Cl.sub.2, and the combined organic layer was dried
(anhydrous Na.sub.2SO.sub.4), filtered, and concentrated by rotary
evaporation to yield a canary yellow foamy solid (0.377 g) of
N4-[2-(1H-indol-2-yl)-phenyl]phthalamic acid. MS (M+H.sup.+: calcd
357; found 357).
[0385] A 5-mL reaction vial with a stirring vane and a teflon
stopper was charged with N-(2-(1H-indol-2-yl)-phenyl)phthalamic
acid, (140 mg; 0.393 mmol) and 0.500 mL of quinoline. To the
solution, which was a dark brown-black, was added zinc acetate
dihydrate (98.0 mg; 0.464 mmol; 1.16 equiv) and the resulting
solution was heated to 120.degree. C. for ca. 2 h. On adding 1 mL
of ethyl acetate, a light tan solid resulted. The solid was washed
with 4.times.10 mL 1 M HCl, then with 10 mL ethylacetate-hexane
(1:1), followed by 10 mL ethyl acetate. The solid was dried in a
vacuum dessicator over phosphorus pentoxide to yield 80.1 mg (71%)
of a light tan solid. MS (M+H.sup.+: calcd 339; found 339).
Pteridine, and Substituted Pteridine Syntheses
##STR00085##
[0386] Experimental Procedure
6,7-(4,4'-Dihydroxyphenyl)-pteridin-4-yl-3-morpholin-4-yl-propyl)-amine
hydrochloride salt
##STR00086##
[0388] 1.19 g (3.59 mmol) of
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine was dissolved in 10 mL
of N-(3-aminopropyl)morpholine and 0.697 g (7.18 mmol, 2.0 eq.) of
sulfamic acid was added. The reaction mixture was heated at
160.degree. C. for 18 hrs. Then it was cooled down to r.t., diluted
with 20 mL of methanol and added dropwise to 1 L of diethyl ether.
The resulting oil was purified by prep-HPLC, fractions were
collected and solvent was removed in vacuo to give red oily
residue, which was dissolved in 20 mL of methanol. 5 g of Amberlite
chloride-exchange resin was added to the methanol solution. The
reaction mixture was left to stir at r.t. overnight, then it was
filtered and resin was washed with methanol. The methanol washes
were combined, solvent was removed in vacuo. The resulting residue
was re-dissolved in 2 mL of methanol and added dropwise to 45 mL of
diethyl ether. The formed bright-yellow precipitate was centrifuged
down, washed with 40 mL of diethyl ether twice and dried in vacuo
to give 281.0 mg (26.2% overall) of the product as a yellow solid.
Mass-spec [ES+]=459.2. 100% purity by LC/MS (230 DAD). .sup.1H NMR
(MeOH-d4) 2.28-2.31 (2H, m), 3.14-3.17 (2H, m), 3.30-3.35 (2H, m),
3.51-3.53 (2H, m), 3.80-3.84 (2H, m), 3.97-4.00 (2H, m), 4.04-4.06
(2H, m), 6.77-6.82 (4H, dd), 7.49-7.54 (4H, dd), 8.84 (1H, s).
Acetic acid 4-[7-(4-acetoxy-phenyl)-4-amino-pteridin-6-yl]-phenyl
ester
##STR00087##
[0390] 662.6 mg (2.0 mmol) of
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine was dissolved in 20 ml
of trifluoroacetic acid. 1.0 mL (14.06 mmol, 7.0 eq) of acetyl
chloride was added via syringe to this mixture. Upon heating to
80.degree. C. bubbling of the reaction mixture and evolution of HCl
gas was observed. The reaction mixture was heated at 80.degree. C.
for 40 min, at which point LC/MS indicated a complete conversion of
the starting material to the di-acetate. Solvent was removed in
vacuo to give bright-yellow oil, which upon standing solidified. 40
mL of diethyl ether was added, the solid was crushed with spatula,
centrifuged down, washed with 45 mL of diethyl ether twice and
dried in vacuo to give 1.034 g (97.7%) of the product as a
light-yellow solid. 97.5% purity by LC/MS (230 DAD). Mass-spec
[ES-F]=416.5. .sup.1H NMR (DMSO-d6) 2.280 (3H, s), 2.284 (3H, s),
7.16-7.21 (4H, dd), 7.56-7.62 (4H, dd), 8.80 (1H, s), 9.46 (1H,
br.s), 9.52 (1H, br.s).
Acetic acid
4-[2-(4-acetoxy-phenyl)-6-amino-pyrido[2,3-b]pyrazin-3-yl]-phenyl
ester
##STR00088##
[0392] 201.0 mg (0.5 mmol) of
2,3-bis(4-hydroxyphenyl)-pyrido[2,3-b]pyrazin-6-ylamine was
dissolved in 10 ml of trifluoroacetic acid. 0.355 mL (5.0 mmol,
10.0 eq) of acetyl chloride was added via syringe to this mixture.
Upon heating to 80.degree. C. bubbling of the reaction mixture and
evolution of HCl gas was observed. The reaction mixture was heated
at 80.degree. C. for 1 hr, at which point LC/MS indicated a
complete conversion of the starting material to the di-acetate.
Solvent was removed in vacuo to give brown solid. The solid was
dissolved in 3.0 mL of methanol and this solution was added to 40
mL of diethyl ether. Upon standing for about an hour a brown
precipitate was formed. It was centrifuged down, washed with 45 mL
of diethyl ether twice and dried in vacuo to give 191.9 mg (79.0%)
of the product as a light-brown solid. 98% purity by LC/MS (230
DAD). Mass-spec [ES+]=415.5. .sup.1H NMR (MeOH-d4) 2.28 (6H, s),
7.10-7.12 (4H, d), 7.24-7.26 (1H, d), 7.48-7.50 (2H, d), 7.54-7.56
(2H, d), 8.24-8.26 (1H, d).
Synthesis of 4-substituted 6-phenyl-pteridin-4-yl-amines
##STR00089##
[0393] General Procedure
[0394] 0.55 mmol of amine was suspended in 4 mL of acetic acid. The
mixture was brought to reflux and 0.5 mmol of
N'-(3-cyano-5-phenyl-pyrazin-2-yl)-N,N'-dimethyl-formamidine was
added to the solution. The reaction was refluxed for 2-5 hours. The
progress of the reaction was monitored by LC/MS. After the reaction
had completed, the reaction mixture was cooled down to ambient
temperature and acetic acid was removed in vacuo. 5 mL of methanol
was added to the resulting residue and it was crushed with a
spatula into a fine suspension. The suspension was added to 45 mL
of diethyl ether. The solid was centrifuged down, washed with 45 mL
of diethyl ether twice and dried in vacuo to give the product as a
solid.
(3,4-Dimethoxy-phenyl)-(6-phenyl-pteridine-4-yl)-amine
##STR00090##
[0396] 95.7% yield. 100% purity by LC/MS (230 DAD). Mass-spec
[ES+]=360.9. .sup.1H NMR (DMSO-d6) 3.79 (3H, s), 3.81 (3H, s),
7.02-7.03 (1H, d), 7.56-7.63 (5H, m), 8.58-8.60 (2H, m), 8.71 (1H,
s), 9.80 (1H, s), 10.27 (1H, s).
(3-Chloro-4,6-dimethoxy-phenyl)-(6-phenyl-pteridin-4-yl)-amine
##STR00091##
[0398] 96% purity by LC/MS (230 DAD). Mass-spec [ES+]=394.9.
.sup.1H NMR (DMSO-d6) 3.92 (3H, s), 3.97 (3H, s), 6.96 (1H, s),
7.59-7.65 (3H, m), 8.29 (1H, s), 8.42-8.43 (2H, d), 8.74 (1H, s),
9.80 (1H, s), 9.89 (1H, s).
(3-Hydroxy-4-methoxy-phenyl)-(6-phenyl-pteridin-4-yl)-amine
##STR00092##
[0400] 79.5% yield. 100% purity by LC/MS (230 DAD). Mass-spec
[ES+]=346.9. .sup.1H NMR (DMSO-d6) 3.79 (3H, s), 6.97-6.98 (1H, d),
7.29-7.31 (1H, dd), 7.46-7.47 (1H, d), 7.58-7.62 (3H, m), 8.58-8.60
(2H, m), 8.69 (1H, s), 9.15 (1H, s), 9.78 (1H, s), 10.2 (1H,
s).
(4-Hydroxy-phenyl)-(6-phenyl-pteridin-4-yl)-amine
##STR00093##
[0402] 86.0% yield. 98% purity by LC/MS (230 DAD). Mass-spec
[ES+]=316.8. .sup.1H NMR (DMSO-d6) 6.82-6.84 (2H, d), 7.57-7.62
(3H, m), 7.65-7.67 (2H, d), 8.58 (2H, m), 8.63 (1H, s), 9.45 (1H,
s), 9.78 (1H, s), 10.26 (1H, s).
(2,5-Dimethyl-4-hydroxy-phenyl)-(6-phenyl-pteridin-4-yl)-amine
##STR00094##
[0404] 76.8% yield. 100% purity by LC/MS (230 DAD). Mass-spec
[ES+]=344.9. .sup.1H NMR (DMSO-d6) 2.12 (6H, s), 6.73 (1H, s), 7.12
(1H, s), 7.55-7.60 (3H, m), 8.54 (1H, s), 8.57-8.58 (2H, m), 9.29
(1H, s), 9.78 (1H, s), 10.16 (1H, s).
2-Hydroxy-5-(6-phenyl-pteridin-4-ylamino)-benzenesulfonic acid
##STR00095##
[0406] 70.1% yield. 83% purity by LC/MS (230 DAD). Mass-spec
[ES+]=396.8. .sup.1H NMR (DMSO-d6) 7.17-7.19 (1H, dd), 7.58-7.63
(3H, m), 7.80-7.82 (1H, dd), 7.993-7.999 (1H, d), 8.61-8.63 (2H,
m), 8.73 (1H, s), 9.80 (1H, s), 10.51-10.53 (3H, m).
2-Diethylaminomethyl-4-(6-phenyl-pteridin-4-ylamino)-phenol
##STR00096##
[0408] 94.3% yield. 98.8% purity by ELSD. Mass-spec [ES+]=402.0.
.sup.1H NMR (DMSO-d6) 1.28-1.31 (6H, t), 3.11-3.16 (4H, m),
4.25-4.26 (2H, d), 7.07-7.09 (1H, d), 7.58-7.63 (3H, m), 7.75-7.77
(1H, dd), 7.89-7.90 (1H, d), 8.57-8.59 (2H, m), 8.67 (1H, s), 9.81
(1H, s), 10.39 (1H, s), 10.5 (1H, s)
5-(6-Phenyl-pteridin-4-ylamino)-quinolin-8-ol hydrochloride
salt
##STR00097##
[0410] 79.9% yield. 85% purity by LC/MS (230 DAD). Mass-spec
[ES+]=367.7. .sup.1H NMR (DMSO-d6) 7.39-7.40 (1H, m), 7.61-7.72
(3H, m), 7.73-7.77 (2H, m), 8.60-8.67 (4H, m), 9.01-9.02 (1H, m),
9.92 (1H, s), 11.58 (1H, br.s.)
Benzyl-(6-phenyl-pteridin-4-yl)-amine
##STR00098##
[0412] 50.5% yield. 95.2% purity by LC/MS (230 DAD). Mass-spec
[ES+]=314.2. .sup.1H NMR (MeOH-d4) 4.87 (2H, s), 7.24-7.26 (1H, m),
7.30-7.33 (2H, m), 7.43-7.44 (2H, m), 7.51-7.54 (3H, m), 8.30-8.32
(2H, m), 8.58 (1H, s), 9.56 (1H, s).
4-[(6-phenyl-pteridin-4-ylamino)-methyl]-benzene-1,2-diol
##STR00099##
[0414] 39.8% yield. 100% purity by LC/MS (230). Mass-spec
[ES+]=346.2. .sup.1H NMR (DMSO-d6) 5.56 (2H, s), 6.68-6.70 (1H, d),
6.75-6.77 (1H, dd), 6.875-6.879 (1H, d), 7.62-7.64 (3H, m),
8.53-8.55 (2H, m), 8.97 (1H, s), 9.12 (1H, s), 9.24 (1H, s), 9.89
(1H, s), 10.48 (1H, s), 10.54 (1H, s).
Indan-2-yl-(6-phenyl-pteridin-4-yl)-amine
##STR00100##
[0416] 53.9% yield. 96.6% purity by LC/MS. Mass-spec [ES+]=340.2.
.sup.1H NMR (DMSO-d6) 3.21-3.26 (2H, dd), 3.35-3.40 (2H, dd),
5.13-5.18 (1H, m), 7.17-7.19 (2H, m), 7.25-7.27 (2H, m), 7.55-7.59
(3H, m), 8.47-8.49 (2H, m), 8.65 (1H, s), 8.94-8.96 (1H, d), 9.72
(1H, s).
2-(3,4-Dimethoxy-phenyl)-ethyl]-(6-phenyl-pteridin-4-yl)-amine
##STR00101##
[0418] 66.5% yield. 95.5% purity by LC/MS (230 DAD). Mass-spec
[ES+]=388.2. .sup.1H NMR (MeOH-d4) 2.98-3.01 (2H, t), 3.76 (3H, s),
3.78 (3H, s), 3.90-3.93 (2H, t), 6.85-6.88 (2H, m), 6.93-6.93 (1H,
m), 7.55-7.57 (3H, m), 8.27-8.29 (2H, m), 8.58 (1H, s), 9.56 (1H,
s)
Synthesis of 4-substituted 7-phenyl-pteridin-4-yl-amines
##STR00102##
[0420] 1N aqueous NaOH was added to a suspension of 1.33 g (5.95
mmol) of 4,5,6-triaminopyrimidine sulfate in 20 mL of water until
pH reached 8. To this solution was added a solution of 1.0 g (5.95
mmol) of 4-hydroxyphenylglyoxal in 20 mL of methanol. The reaction
mixture was left to stir at ambient temperature for 18 hrs.
Formation of a yellow precipitate was observed. It was collected,
washed with 20 mL of water, 20 mL of methanol, 45 mL of diethyl
ether 3 times and dried in vacuo to give 1.513 g of the product as
a light-yellow solid. 100% yield. 97.5% purity by LC/MS (230 DAD).
Mass-spec [ES+]=. .sup.1H NMR (DMSO-d6) 6.95-6.98 (2H, d), 8.31
(1H, br.s.), 8.19 (1H, br.s.), 8.21-8.24 (2H, d), 8.51 (1H, s),
9.34 (1H, s).
General Procedure
[0421] 239.2 mg (1.0 mmol) of 4-(4-amino-pteridin-7-yl)-phenol was
suspended in 3 mL of amine and 194.2 mg (2.0 mmol) of sulfamic acid
was added to this mixture. The reaction mixture was heated at
160-180.degree. C. for 18 hrs. Then it was cooled down to ambient
temperature and dissolved in 5-10 mL of methanol. Methanol solution
was added dropwise to 45 mL of diethyl ether, the mixture was
vortexed and centrifuged down. Solvent was decanted and the residue
was purified by prep-HPLC.
4-(4-Benzylamino-pteridin-7-yl)-phenol
##STR00103##
[0423] 79% yield. 98.5% purity by LC/MS (230 DAD). Mass-spec
[ES+]=330.2. .sup.1H NMR (DMSO-d6) 4.77-4.78 (2H, d), 6.97-6.98
(2H, d), 7.24-7.26 (1H, m), 7.30-7.33 (2H, m), 7.43-7.44 (2H, m),
8.23-8.24 (2H, d), 8.58 (1H, s), 9.37 (1H, s).
Substituted (6-phenyl-5,6,7,8-tetrahydro-pteridin-4-yl)-amines and
(7-phenyl-5,6,7,8-tetrahydro-pteridin-4-yl)-amines
##STR00104##
[0424] General Procedure
[0425] To a stirred solution of the pteridine (5.0 mmol) in 15 mL
of dry methanol was added sodium borohydride (5 mmol) at room
temperature. The reaction mixture was stirred for 30 min and then
neutralized with acetic acid. Solvent was removed in vacuo and the
residue was washed with water, cold methanol, diethyl ether and
dried in vacuo. The resulting solid was purified by reverse phase
prep-HPLC.
6,7-disubstituted pteridines; Method A
##STR00105##
[0426] Method B
[0427] The pyridine or pyrimidine is made into the free base with
sodium carbonate, sodium bicarbonate or sodium hydroxide using
solid or solution by using the correct amount in equivalents to
neutralize the acid or by adjusting the pH to neutral to slightly
basic (ca. 7-9). The benzil or glyoxal is added and the solution is
heated for 1 h-5 h. The free base formed precipitates out of
solution and is washed successively with water, methanol and then
ether. The solid is vacuum dessicator dried.
[0428] This reaction was carried out by method A by using 23.5 mg
of the pyrimidine and 22.5 mg of pyridyl. The reaction mixture was
heated for 1 h. The product was precipitated into 5 mL of 1:1
EtOAc-ether, filtered and washed with 50 mL of ether. M+H calcd and
found 400.
6,7-bis(3-hydroxyphenyl)-pteridine-2,4,-diamine
[0429] A 5-mL reaction vial with a stirring vane and a teflon cap
was charged with 3,3'-dihydroxybenzil (Midori Kagaku Co Ltd; 121
mg; 0.500 mmol) and 0.700 mL of m-cresol (Acros) which gives a
dull-yellow solution on warming to ca. 50.degree. C. The clear
solution is treated with 2,4,5,6-tetraminopyrimidine sulfate
(Aldrich; 119 mg; 0.500 mmol; 1.00 equiv) which is insoluble in the
reaction solution at room temperature and goes into solution on
heating to ca. 200.degree. C. to give an almost completely
homogeneous dark greenish solution in about 30 min-45 min. Heating
between 200.degree. C. and 220.degree. C. for an additional 1.5 h,
followed by cooling to room temperature, and precipitation by
pouring into 40 mL of anhydrous diethyl ether resulted in a
greenish-yellow precipitate. The solid was centrifuged, the
supernatant decanted, the solid precipitate was washed with
5.times.40 mL of diethyl ether and dried in a vacuum dessicator to
yield 0.275 g (124%).sup.1 of a yellow-green solid. The only
obvious major impurity is the reaction solvent, m-cresol. MS
(M+H.sup.+: calcd 347; found 347).
[0430] In case purified
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine is required, the
crude 3-[2,4-diamino-6-(3-hydroxyphenyl)pteridin-7-yl]phenol may be
dissolved in methanol, and an aqueous solution of 2.0 equiv. -2.2
equiv. of sodium bicarbonate (or excess sodium bicarbonate) may be
added to neutralize the acid making sure the pH is between 6 and 8
to ensure free-base. The free-base precipitates out of the
methanol-water mixture within a few seconds. In case, precipitation
does not occur, excess methanol ensures precipitation. The
yellowish solid may be isolated and washed with acetonitrile-water
or isopropanol-water mixtures and then with methanol-ether, and
then ether (.times.3). The product is dried and stored as the free
base, 6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine.
[0431] In case the purified sulfate is required, the free base is
protonated in MeOH by adding a conc. aqueous sulfuric acid (1.0
equiv) to a slurry of the compound in MeOH. The homogeneous
protonated product is precipitated out by adding ether to the
methanol.
6-pyridin-2-yl-7-pyridin-3-ylpteridin-4-amine sulfate salt
##STR00106##
[0433] A 5-mL reaction vial with a stirring vane and a teflon cap
was charged with pyridyl (22.5 mgl) and 0.500 mL of m-cresol
(Acros) which gives a dull-yellow solution on warming to ca.
50.degree. C. The clear solution is treated with
2,4,5-triaminopyrimidine sulfate (Aldrich; 23.5 mg) which is
insoluble in the reaction solution at room temperature and goes
into solution on heating to ca. 200.degree. C. to give an almost
completely homogeneous dark solution in about 30 min-45 min.
Heating between 200.degree. C. and 220.degree. C. for an additional
0.5 h, followed by cooling to room temperature, and precipitation
by pouring into 40 mL of anhydrous diethyl ether resulted in a dull
yellow precipitate. The solid was centrifuged, the supernatant
decanted, the solid precipitate was washed with 4.times.40 mL of
diethyl ether and dried in a vacuum dessicator to yield a yellow
solid. MS (M+H.sup.+: calcd 302; found 302).
6,7-bis(3,4-dihydroxyphenyl)pteridine-2,4-diol
##STR00107##
[0435] A 5-mL reaction vial with a stirring vane and a teflon cap
was charged with 3,3',4,4'-tetrahydroxybenzil (137 mg; 0.500 mmol)
and 1.00 mL of m-cresol (Acros) which gives a yellow-brown slurry
warming to ca. 50.degree. C. The suspension is treated with sulfate
5,6-diamino-2,4-dihydroxypyrimidine sulfate (120 mg; 0.500 mmol;
1.00 equiv) which is insoluble in the reaction solution at room
temperature and goes into solution on heating to ca. 200.degree. C.
to give homogeneous dark solution. Heating between 200.degree. C.
and 220.degree. C. for an additional 2 h, followed by cooling to
room temperature, and precipitation by pouring into 40 mL of
anhydrous diethyl ether resulted in a light yellow precipitate. The
solid was centrifuged, the supernatant decanted, the solid
precipitate was washed with 4.times.40 mL of diethyl ether and
dried in a vacuum dessicator to yield a yellow solid. MS
(M+H.sup.+: calcd 381; found 381).
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine dihydrochloride
salt
##STR00108##
[0437] A 125-mL amber-bottle with a stirring bar and a septum was
charged with crude 6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine
(135 mg; 0.304 mmol) and 5 mL of methanol. To the resulting dark
brownish-green solution was added Amberlite (co resin (GFS
Chemical; 5.20 g). The heterogeneous mixture was stirred gently for
ca. 16 h. with an apparent visual lightening of the solution. The
solution was filtered to remove the resin beads, which were rinsed
with 5.times.8 mL of MeOH. The light brown solution was
concentrated on a rotary evaporator to yield 133 mg of dark brown
oil. The oil was redissolved in ca. 2 mL of MeOH, and added to 40
mL of diethyl ether to yield a flocculent yellow precipitate that
was isolated by centrifuging and decanting the supernatant. The
solid was washed with 4.times.40 mL of diethyl ether, and dried in
a vacuum dessicator to yield a greenish-yellow product (94.0 mg;
0.246 mmol; 81% for two steps). 98% purity by LC/MS (230 DAD).
Mass-spec [ES.sup.+]=347.7. .sup.1H NMR (DMSO-d6) 6.78-6.87 (4H,
m), 6.92-6.95 (2H, m), 7.12-7.16 (2H, m), 7.82 (1H, br.s), 8.68
(1H, br.s), 9.15 (1H, s), 9.25 (1H, s), 9.58 (1H, s), 9.72 (1H, s).
C, N analysis: C.sub.18H.sub.16Cl.sub.2N.sub.6O.sub.2 (Calcd.: C,
51.56; N, 20.04. Found: C, 51.64; N, 19.93).
Method B
##STR00109##
[0438] 6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine
##STR00110##
[0440] 4.76 g (20.0 mmol) of 2,4,5,6-tetraminopyrimidine sulfate
was added in small portions to a solution of 3.36 g (40.0 mmol)
sodium bicarbonate in 100 mL of water with vigorous stirring. A
brisk evolution of CO.sub.2 gas was observed. The resulting
suspension was heated to 80.degree. C. and 4.84 g (20.0 mmol) of
3,3'-dihydroxybenzil was added to the mixture. The reaction mixture
was refluxed for 3 hours, at which point a bright-yellow
precipitate was formed in abundance.
[0441] The precipitate was filtered, washed with water, then with
methanol, followed by diethyl ether and dried in vacuo to give 6.46
g (93.3% yield) of a bright-yellow solid. 98.10% purity by LC/MS
(230 DAD). Mass-spec [ES.sup.+]=347.7. .sup.1H NMR (DMSO-d6) 6.64
(2H, br.s.), 6.69-6.82 (4H, m), 6.86-6.89 (2H, m), 7.06-7.11 (2H,
m), 7.57 (1H, br.s), 7.65 (1H, br.s), 9.38 (1H, s), 9.49 (1H,
s).
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine methanesulfonate
salt
##STR00111##
[0443] 2.66 g (7.68 mmol) of
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine was added to a
solution of 1.55 g (16.13 mmol) of methanesulfonic acid in 20 mL of
MeOH with stirring. Pteridine immediately dissolved to give a
dark-greenish solution. The reaction mixture was stirred for 30 min
and then added dropwise to 400 mL of diethyl ether with vigorous
stirring. The formed yellow precipitate was collected, washed
repeatedly with ether and dried in vacuo to give 3.36 g (99.1%
yield) of the product as a light-yellow powder. 95.5% purity by
LC/MS (230 DAD). Mass-spec [ES+]=347. .sup.1H NMR (MeOH-d4) 2.71
(3H, s), 6.80-6.85 (2H, m), 6.90-6.92 (2H, m), 6.95 (1H, m), 7.00
(1H, m), 7.12-7.16 (2H, m).
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine dihydrobromide
salt
[0444] The salt is made by making a HBr containing solution of
methanol using methanol and acetyl bromide (10 equiv-12 equiv) at
-78.degree. C., and adding the free base to this solution so that
the resulting solution concentration is below 0.4 M. The light
yellow solution is stirred for ca. 30 min-60 min, concentrated by
rotary evaporation to a yellow solid and then washed with ether, or
with ether-hexanes, and dried in a vacuum dessicator
##STR00112##
[0445] 98.8% Purity by LC/MS (230 DAD). Mass-spec [ES.sup.+]=347.
.sup.1H NMR (MeOH-d4) 6.81-6.86 (2H, m), 6.92-6.95 (2H, m),
6.96-7.01 (2H, m), 7.13-7.18 (2H, m). Elemental analysis; calcd: C,
42.54; H, 3.17; N, 16.54. found: C, 43.11; H, 3.47; N, 16.47
6,7-bis(3-hydroxyphenyl)-pteridin-4-ylamine
##STR00113##
[0447] 2.23 g (10.0 mmol) of 4,5,6-triaminopyrimidine sulfate was
added in small portions to a solution of 1.68 g (20.0 mmol) sodium
bicarbonate in 50 mL of water with vigorous stirring. A brisk
evolution of CO.sub.2 gas was observed. The resulting suspension
was heated to 80.degree. C. and 2.42 g (10 mmol) of
3,3'-dihydroxybenzil was added to the mixture. The reaction mixture
was refluxed for 1 hour, during which time the starting materials
completely dissolved and the product precipitated out as a
light-yellow solid.
[0448] The precipitate was collected, washed with water, then with
methanol, followed by diethyl ether and dried in vacuo to give 3.14
g (94.8% yield) of the product as a light-yellow solid. 100% purity
by LC/MS (230 DAD). Mass-spec [ES.sup.+]=332.8. .sup.1H NMR
(DMSO-d6) 6.77-6.83 (3H, m), 6.91-6.92 (1H, d), 6.90-6.99 (2H, m),
7.11-7.15 (2H, m), 8.17 (1H, br.s), 8.25 (1H, br.s.), 8.56 (1H, s),
9.55 (2H, br.s).
6,7-bis(3-hydroxyphenyl)-pteridin-4-ylamine hydrochloride salt
##STR00114##
[0450] 4.4 g (13.27 mmol) of
6,7-bis(3-hydroxyphenyl)-pteridin-4-ylamine was suspended in 35 ml
of MeOH. A solution of 2.61 g of aq. HCl (26.55 mmol, 12.1 N) in 5
mL of MeOH was added to the suspension. The reaction mixture became
homogeneous within 5 min of stirring. It was left to stir for 30
min and then added dropwise to 400 mL of diethyl ether with
vigorous stirring. The resulting precipitate was collected, washed
repeatedly with ether and dried in vacuo to give 4.62 g (94.7%
yield) of the product as a bright-yellow solid. 98.3% purity by
LC/MS (230 DAD). Mass-spec [ES.sup.+]=332.8. .sup.1H NMR (MeOH)
6.88-6.90 (2H, m), 6.99-7.02 (2H, m), 7.04-7.08 (2H, m), 7.17-7.20
(2H, m), 8.79 (1H, s).
6,7-bis(3-hydroxyphenyl)-pteridin-4-ylamine methanesulfonate
salt
##STR00115##
[0452] 1.308 g (13.63 mmol) of methanesulfonic acid in 10 mL of
MeOH was added to the suspension of 2.15 g (6.48 mmol) of
6,7-bis(3-hydroxyphenyl)-pteridin-4-ylamine in 10 mL of MeOH. The
mixture became homogeneous and orange-red in color. It was stirred
for 30 min and then added dropwise to 400 mL of diethyl ether with
vigorous stirring. The formed yellow precipitate was collected,
washed with diethyl ether and dried in vacuo to give 2.69 g (97.11%
yield) of the product as a light-yellow powder. Mass-spec
[ES.sup.+]=332.8. .sup.1H NMR (MeOH-d4) 2.70 (3H, s), 6.86-6.90
(2H, m), 6.99-7.01 (2H, m), 7.04-7.08 (2H, m), 7.16-7.21 (2H, m),
8.80 (1H, s).
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine
##STR00116##
[0454] 1.5 mmol of the sulfate salt
(6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine sulfate salt as 1:1
complex with m-cresol) was dissolved in 10 mL of 1:1 solution of
MeOH/H.sub.2O. 2.0 eq. of solid NaHCO.sub.3 were added to this
solution. A brisk evolution of CO.sub.2 was observed and a
light-yellow precipitate started to form in .about.10-15 min of
stirring. The mixture was left to stir overnight and a yellow
precipitate was formed in abundance. 20 mL of water was added, the
formed precipitate was filtered, washed twice with water to remove
Na.sub.2SO.sub.4, washed with cold MeOH, washed repeatedly with
Et.sub.2O and dried in vacuo to give the product in 81.3% yield
over two steps (reaction in m-cresol and free base synthesis).
95.5% purity by LC/MS (230 DAD). Mass-spec [ES.sup.+]=332.8.
.sup.1H NMR (DMSO-d6) 6.72-6.76 (4H, dd), 7.35-7.42 (4H, dd), 8.06
(1H, br.s), 8.14 (1H, br.s), 8.50 (1H, s), 9.77 (1H, br.s), 9.87
(1H, br.s)
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine sulfate salt
##STR00117##
[0456] 1.97 g of 6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine was
added to a solution of 0.585 g of concentrated sulfuric acid in 50
mL of MeOH. The homogeneous mixture was left to stir at ambient
temperature for 2 hours, then it was added dropwise to 400 mL of
diethyl ether. The formed orange precipitate was collected, washed
repeatedly with ether and dried in vacuo to give 2.36 g (92.5%
yield) of the product as a light-orange fluffy powder. 100% purity
by LC/MS (230 DAD). Mass-spec [ES.sup.+]=332.8. .sup.1H NMR
(MeOH-d4) 6.77-6.80 (4H, m), 7.48-7.53 (4H, m), 8.73 (1H, s).
.sup.1H NMR (DMSO-d6) 6.76-6.81 (4H, dd), 7.41-7.47 (4H, dd), 8.84
(1H, s), 9.85 (1H, s), 10.01 (1H, s), 9.94 (1H, br.s), 10.15 (1H,
br.s).
6,7-bis(3,4-dihydroxyphenyl)-pteridine-2,4-diamine
##STR00118##
[0458] 105.0 mg (0.253 mmol) of
6,7-bis(3,4-dihydroxyphenyl)-pteridine-2,4-diamine dihydrochloride
salt was dissolved in 3 mL of water and 42.53 mg of solid NaHCO3
was added to this solution. The reaction mixture was stirred for 30
min. A slurry of yellow precipitate was formed, it was centrifuged
down and solvent was decanted. The dark-yellow residue was
dissolved in 3 mL of MeOH and added dropwise to 40 mL of diethyl
ether. The formed yellow precipitate was collected, washed with
ether and dried in vacuo to give 92.5 mg (96.5% yield) of the
product as a yellow, fluffy powder. 97% purity by LC/MS (230 DAD).
Mass-spec [M+H.sup.+]=379.3. .sup.1H NMR (MeOH-d4) 6.68-6.73 (2H,
dd), 6.79-6.81 (1H, dd), 6.84-6.86 (1H, dd), 6.93 (1H, d), 7.03
(1H, d).
6,7-bis(3,4-dihydroxyphenyl)-pteridine-2,4-diamine dihydrochloride
salt
##STR00119##
[0460] Mass-spec [ES.sup.+]=379.8. .sup.1H NMR (MeOH-d4) 6.70 (1H,
d), 6.75 (1H, d), 6.88 (1H, dd), 6.93 (1H, dd), 6.95 (1H, d), 7.08
(1H, d).
6,7-bis(3,4-dihydroxyphenyl)-pteridin-4-ylamine hydrochloride salt
or 4-[4-amino-6-(3,4-dihydroxyphenyl)pteridin-7-yl]benzene-1,2-diol
hydrochloride salt
##STR00120##
[0462] A 5-mL reaction vial with a stirring vane and a teflon cap
was charged with 3,3',4,4'-tetrahydroxybenzil (Midori Kagaku Co
Ltd; 548 mg; 2.00 mmol), 4,5,6-triaminopyrimidine sulfate and 3.00
mL of m-cresol. The heterogeneous mixture was heated, it first goes
orange while dissolving at ca. 150.degree. C. and then on heating
at 200.degree. C.-220.degree. C. for ca. 2 h goes to a dark
blood-red solution. The clear solution is heated for an additional
30 min, followed by cooling to room temperature, and precipitation
by pouring into 40 mL of anhydrous diethyl ether resulted in a dark
red-orange precipitate. The solid was centrifuged, washed with
5.times.40 mL of diethyl ether and dried in a vacuum dessicator to
yield 1.20 g (128%).sup.1 of an orange-red solid. The only obvious
major impurity is the reaction solvent, m-cresol.
[0463] Mass-spec [ES+]=364.8. .sup.1H NMR (MeOH-d4) 6.73 (1H, d),
6.78 (1H, d), 7.00-7.02 (2H, dd), 7.07 (1H, d), 7.16 (1H, d). 8.71
(1H, s).
6,7-bis(3,4-dihydroxyphenyl)-pteridin-4-ylamine or
4-[4-amino-6-(3,4-dihydroxyphenyl)pteridin-7-yl]benzene-1,2-diol
##STR00121##
[0465] Mass-spec [ES.sup.+]=364.8. .sup.1H NMR (MeOH-d4) 6.70-6.75
(2H, dd), 6.91-6.95 (2H, dd), 7.03 (1H, d), 7.12 (1H, d), 8.49 (1H,
s). NMR (DMSO-d6) 6.63-6.68 (2H, dd), 6.74-6.76 (1H, dd), 6.85-6.87
(1H, dd), 7.00 (1H, d), 7.06 (1H, d), 7.93 (2H, br.s), 8.47 (1H,
s).
6,7-bis(3,4-dihydroxyphenyl)-pteridin-4-ylamine methanesulfonate
salt or
4-[4-amino-6-(3,4-dihydroxyphenyl)pteridin-7-yl]benzene-1,2-diol
methanesulfonate salt
##STR00122##
[0467] 98.07% purity by LC/MS (230 DAD). Mass-spec
[ES.sup.+]=364.8. .sup.1H NMR (MeOH-d4) 2.69 (3H, s), 6.73-6.79
(2H, dd), 7.00-7.04 (2H, dd), 7.08 (1H, d), 7.17 (1H, d), 8.81 (1H,
s).
4-(2,4-diaminopteridin-6-phenol
##STR00123##
[0469] A 50-mL recovery flask fitted with a stirring bar, a reflux
condensor and a heating mantle was charged with 1 mmol of each of
hydroxylamine hydrochloride and 4-hydroxyphenylglyoxal. The
substances were dissolved in methanol (5 mL). To this yellow
solution was added the 2,4,5,6-tetraminopyrimidine sulfate and 20
mL of water. The heterogeneous solution was heated to reflux for 2
h. A yellow precipitate that was formed. The solution was cooled,
the reaction mixture was made slightly basic NaOH (4 M, aqueous) to
a pH of ca. 8. The precipitated free base was isolated and washed
sequentially with water (2.times.40 mL), methanol (1.times.40 mL)
and ether (1.times.40 mL) and drying in a vacuum dessicator.
##STR00124##
[0470] A 5-mL reaction vial with a stirring vane and a teflon cap
was charged with benzil (420 mg; 2.00 mmol) and 2.00 mL of m-cresol
(Acros) which gives a dull-yellow solution on warming to ca.
50.degree. C. The clear solution is treated with
5,6-diamino-2,4-dihydroxypyrimidine sulfate (Aldrich; 482 mg; 2.00
mmol; 1.00 equiv) which is insoluble in the reaction solution at
room temperature and goes into solution on heating to ca.
200.degree. C. to give an almost completely homogeneous dark
solution in about 30 min-45 min. Heating between 200.degree. C. and
220.degree. C. for an additional 1.5 h, followed by cooling to room
temperature, and precipitation by pouring into 40 mL of anhydrous
diethyl ether resulted in a dull yellow precipitate. The solid was
centrifuged, the supernatant decanted, the solid precipitate was
washed with 4.times.40 mL of diethyl ether and dried in a vacuum
dessicator to yield 960 mg (99%) of a yellow solid. MS (M+H.sup.+:
calcd 317; found 317).
4-(2,4-Diamino-pteridin-6-yl)-phenol
[0471] (M+H)+ calcd and found 255; LC (UV-PDA 230 nm) 98% purity.;
.sup.1H NMR (500 MHz; DMSO-d.sub.6): .delta. 9.89 (br s, 1H), 9.24
(s, 1H), 8.15 (d, J=8.5 Hz, 2H), 7.70 (br.s, 1H), 7.65 (br. s, 1H)
6.88 (d, J=8.5 Hz, 2H), 6.57 (br s, 2H)
2,3-Diphenyl-pyrido[3,4-b]pyrazin-8-ylamine hydrochloride salt
##STR00125##
[0473] 60.0 mg (0.37 mmol) of 3,4,5-triaminopyrimidine
hydrochloride and 86.3 mg (0.41 mmol) of benzil were heated at
190.degree. C. in 1.0 mL of m-cresol for 1 hr. Then the mixture was
cooled down to r.t., mixed with 35 mL of diethyl ether. The formed
brown precipitate was collected, washed repeatedly with ether and
dried in vacuo to give 51.1 mg (45.8% yield) of the product as a
brown powder. Mass-spec [M+H.sup.+]=299.2. 1H NMR (MeOH-d4)
7.38-7.41 (3H, m), 7.45-7.49 (3H, m), 7.58-7.60 (2H, m), 7.66-7.68
(2H, m), 8.05 (1H, s), 8.85 (1H, s).
2,3-Bis(4-hydroxyphenyl)-pyrido[3,4-b]pyrazin-8-ylamine
hydrochloride salt
##STR00126##
[0475] 60.0 mg (0.37 mmol) of 3,4,5-triaminopyrimidine
hydrochloride and 99.6 mg (0.41 mmol) of 4,4'-dihydroxybenzil were
heated at 190.degree. C. in 1.0 mL of m-cresol for 1 hr. Then the
mixture was cooled down to r.t., mixed with 35 mL of diethyl ether.
The formed brown precipitate was collected, washed repeatedly with
ether and dried in vacuo to give 91.3 mg (66.6% yield) of the
product as a dark-green powder. Mass-spec [M+H.sup.+]=331.4.
.sup.1H NMR (MeOH-d4) 6.78-6.81 (4H, d), 7.49-7.51 (2H, d),
7.60-7.62 (2H, d), 7.95 (1H, s), 8.71 (1H, s).
2,3-Bis(3,4-dihydroxyphenyl)-pyrido[3,4-b]pyrazin-8-ylamine
hydrochloride salt
##STR00127##
[0477] 60 mg (0.37 mmol) of 3,4,5-triaminopyridine hydrochloride
and 112.6 mg (0.41 mmol) of 3,3',4,4'-tetrahydroxybenzil were
dissolved in 1 mL of m-cresol. The reaction mixture was heated at
190.degree. C. for 1 hr, at which point the mixture became
homogeneous and dark-brown in color. The reaction was cooled to
r.t. and mixed with 35 mL of diethyl ether. The formed brown
precipitate was vortexed, collected, washed repeatedly with diethyl
ether and dried in vacuo to give 111.0 mg (82% yield) of the
product. Mass-spec [M+H.sup.+]=363.2. .sup.1H NMR (MeOH-d4)
6.76-6.78 (2H, d), 6.98-7.00 (1H, dd), 7.11 (1H, dd), 7.13 (1H, d),
7.21 (1H, dd), 7.94 (1H, s), 8.68 (1H, s).
2,3-Bis(3-hydroxyphenyl)-pyrido[3,4-b]pyrazin-8-ylamine
hydrochloride salt
##STR00128##
[0479] 60.0 mg (0.37 mmol) of 3,4,5-triaminopyrimidine
hydrochloride and 99.6 mg (0.41 mmol) of 3,3'-hydroxybenzil were
heated at 190.degree. C. in 1.0 ml of m-cresol for 1 hr. Then the
mixture was cooled down to r.t., mixed with 35 ml of diethyl ether.
The formed brown precipitate was collected, washed repeatedly with
ether and dried in vacuo to give 93.9 mg (68.5% yield) of the
product as a greenish-brown powder. Mass-spec [M+H.sup.+]=331.4.
.sup.1H NMR (MeOH-d4) 6.88-6.91 (2H, m), 6.99-7.01 (1H, m),
7.07-7.10 (2H, m), 7.13-7.14 (1H, m), 7.18-7.22 (2H, m), 8.03 (1H,
s), 8.82 (1H, s).
2,3-bis(3-hydroxyphenyl)-pyrido[2,3-b]pyrazin-6-ylamine
dihydrochloride salt
##STR00129##
[0481] 197.0 mg (1.0 mmol) of 2,3,6-triaminopyrimidine
dihydrochloride and 242.4 mg (1.0 mmol) of 3,3'-dihydroxybenzil
were dissolved in 3.0 mL of 1:1 mixture of dioxane-water. The
reaction mixture was refluxed for 3 hours and then solvent was
removed in vacuo. The resulting greenish solid was dissolved in 3
mL of MeOH and this solution was added to 40 mL of diethyl ether
with vigorous stirring. The formed precipitate was collected,
washed with diethyl ether and dried in vacuo to give 342.9 mg
(85.0% yield) of the product as a light-green powder. 99.0% purity
by LC/MS (230 DAD). Mass-spec [ES.sup.+]=331.8. .sup.1H NMR
(MeOH-d4) 6.83-6.85 (2H, m), 6.88-6.90 (1H, m), 6.95-6.97 (2H, m),
7.02-7.03 (1H, m), 7.14-7.18 (2H, m), 7.36-7.38 (1H, d), 8.43-8.46
(1H, d).
2,3-bis(4-hydroxyphenyl)-pyrido[2,3-b]pyrazin-6-ylamine
dihydrochloride salt
##STR00130##
[0483] 1.97 g (10.0 mmol) of 2,3,6-triaminopyrimidine
dihydrochloride and 2.42 g (10.0 mmol) of 4,4'-dihydroxybenzil were
dissolved in 30 mL of 1:1 mixture of dioxane-water. The reaction
mixture was refluxed for 6 hours and then solvent was distilled
off. The resulting dark-brown solid was suspended in 20 mL of MeOH
and this suspension was added to 400 mL of diethyl ether with
vigorous stirring. The formed dark-brown precipitate was collected,
washed with diethyl ether and dried in vacuo to give 3.35 g (83.1%
yield) of the product as a brown fluffy powder. 92.6% purity by
LC/MS (230 DAD). Mass-spec [ES.sup.+]=331.8. .sup.1H NMR (MeOH-d4)
6.72-5.77 (4H, m), 7.29-7.33 (3H, m), 7.40-7.42 (1H, m), 7.41 (1H,
d), 8.35 (1H, d).
Phosphate ester of 4,4'-dihydroxybenzil
##STR00131##
[0485] A 50-mL one-necked round-bottomed flask with a stirring bar
and a septum was charged with 4,4'-dihydroxybenzil (512 mg; 2.11
mmol; 1.00 equiv) and acetonitrile (8 mL). To this partially
dissolved mixture was added triethylamine (1.06 g; 14.9 mmol; 7.06
equiv), dimethylaminopyridine (DMAP) (478 mg; 3.91 mmol; 1.85
equiv) and dichloromethane (DCM) as co-solvent. The reaction
mixture was stirred for 3 d at room temperature after which it was
concentrated by rotary evaporation to yield a yellow-white slurry.
This oily slurry was partitioned between sodium bicarbonate (satd.
aq) and dichloromethane (DCM). The aqueous layer was rewashed with
2.times.5 mL DCM, followed by extraction of the combined organics
with 10 mL of 1 M HCl. The DCM layer was dried (anhyd. MgSO.sub.4),
filtered and concentrated by rotary evaporation to yield the
desired material as a light yellow slightly viscous oil. The
compound does not require any purification but is easily purified
by column chromatography using DCM-EtOAc (1:1). The
chromatographically purified material is a yellow oil (911 mg;
89%).
[0486] .sup.1H NMR (500 MHz; DMSO-d.sub.6): .delta. 8.01 (d, J=8.6
Hz, 4H), 7.45 (d, J=8.5 Hz, 4H), 4.21-4.18 (m, 8H), 1.28 (app t,
J=5.0 Hz, 12H)
[0487] The compound was made by the method B in the pteridine
synthesis by using the pyrimidine and the phosphate ester of the
4,4'-dihydroxybenzil.
##STR00132##
[0488] The compound was purified by passing through a plug of
silica using ethyl acetate. (M+H)+: calcd. 604; found 604. LC
purity 96% (DAD at 230 nm).
[0489] .sup.1H NMR (500 MHz; DMSO-d.sub.6); .delta. 8.58 (s, 1H),
8.30 (br s, 2H), 7.58 (d, J=6.8 Hz), 7.54 (d, J=6.8 Hz, 2H), 7.23
(d, J=8.8 Hz, 2H), 7.20 (d, J=8.9 Hz, 2H), 4.17-4.14 (m, 8H), 1.26
(app t, J=6.9 Hz, 12H)
[0490] Phosphate Ester Deprotected
[0491] The above diethylester compound was deprotected in
acetonitrile using TMSBr. The reaction was completed by adding
water and then concentration by rotary evaporation and drying of
the solid.
##STR00133##
[0492] .sup.1H NMR (500 MHz; methanol-d.sub.4); .delta. 8.39 (s,
1H), 7.31 (d, J=6.8 Hz, 2H), 7.26 (d, J=6.7 Hz, 2H), 6.31 (app t,
J=6.8 Hz, 4H)
[0493] Phosphate Ester of Pyridopyrazine
##STR00134##
[0494] .sup.1H NMR (500 MHz; DMSO-d.sub.6): .delta. 8.05 (d, J=9.0
Hz, 1H), 7.46 (d, J=8.7 Hz, 2H), 7.43 (d, J=8.6 Hz, 2H), 7.24 (br
s, 2H), 7.17 (app t, J=7.7 Hz, 4H), 7.10 (d, J=9.0 Hz, 1H),
4.17-4.13 (m, 8H), 1.26 (app t, J=5.0 Hz, 12H)
[0495] Phosphate Ester Deprotected
##STR00135##
[0496] This compound was made in a similar fashion to the one
described above.
[0497] .sup.1H NMR (500 MHz; methanol-d.sub.4); .delta. 8.05 (d,
J=9.0 Hz, 1H), 7.46 (d, J=8.7 Hz, 2H), 7.43 (d, J=8.6 Hz, 2H), 7.24
(br s, 2H), 7.17 (app t, J=7.7 Hz, 4H), 7.10 (d, J=9.0, 2H).
[0498] Long Chain Ester of Pteridine
[0499] The benzil was modified using an acid chloride with DMAP as
base in DCM. The modified Benzil was then condensed with the
pyrimidine to yield the product below.
##STR00136##
4-(4-amino-pteridin-7-yl)-benzene-1,2-diol
[0500] This compound is made by stirring a 1:1 ratio of the
appropriate glyoxal with the free base of the pyrimidine in water
at a pH of 7 for ca. 3 h. The product is isolated by filtering the
precipitated free base, washing sequentially with water (2.times.40
mL), methanol (1.times.40 mL) and ether (2.times.40 mL) and drying
in a vacuum dessicator.
##STR00137##
[0501] .sup.1H NMR (500 MHz; DMSO-d.sub.6): .delta. 9.72 (s, 1H),
9.40 (br s, 1H), 9.28 (s, 1H), 8.51 (s, 1H), 8.17 (br s, 1H), 8.12
(br s, 1H), 7.80 (d, J=2.3 Hz, 1H), 7.71 Hz, (dd, J=8.4 Hz, J=2.3
Hz, 1H), 6.92 (d, J=8.3 Hz, 1H).
4-(2,4-diamino-pteridin-7-yl)-benzene-1,2-diol
[0502] This compound is made by stirring a 1:1 ratio of the
appropriate glyoxal with the free base of the pyrimidine in water
at a pH of 7 for ca. 3 h. The product is isolated by filtering the
precipitated free base, washing sequentially with water (2.times.40
mL), methanol (1.times.40 mL) and ether (2.times.40 mL) and drying
in a vacuum dessicator.
##STR00138##
[0503] .sup.1H NMR (500 MHz; DMSO-d.sub.6): .delta. 8.71 (s, 1H),
7.64 (d, J=2.3 Hz, 1H), 7.56-7.53 (br s, 2H), 7.53 (dd, J=8.3 Hz,
2.1 Hz, 1H), 6.84 (d, J=8.3 Hz, 1H), 6.52 (br s, 2 H)
4-(4-amino-pteridin-7-yl)-phenol
[0504] This compound is made by stirring a 1:1 ratio of the
appropriate glyoxal with the free base of the pyrimidine in water
at a pH of 7 for ca. 3 h. The product is isolated by filtering the
precipitated free base, washing sequentially with water (2.times.40
mL), methanol (1.times.40 mL) and ether (2.times.40 mL) and drying
in a vacuum dessicator.
##STR00139##
[0505] .sup.1H NMR (500 MHz; DMSO-d.sub.6): .delta. 10.2 (br s,
1H), 9.34 (s, 1H), 8.52 (s, 1H), 8.23 (d, J=6.8 Hz, 2H), 8.19 (br
s, 1H), 8.13 (br s, 1H), 6.97 (d, J=8.8 Hz, 2H)
4-(2,4-diamino-pteridin-7-yl)-phenol
[0506] This compound is made by stirring a 1:1 ratio of the
appropriate glyoxal with the free base of the pyrimidine in water
at a pH of 7 for ca. 3 h. The product is isolated by filtering the
precipitated free base, washing sequentially with water (2.times.40
mL), methanol (1.times.40 mL) and ether (2.times.40 mL) and drying
in a vacuum dessicator.
##STR00140##
[0507] .sup.1H NMR (500 MHz; DMSO-d.sub.6): .delta. 10.0 (br s,
1H), 8.81 (s, 1H), 8.09 (d, J=8.5 Hz, 2H), 7.62 (br s, 1H), 7.55
(br s, 1H), 6.91 (d, J=8.5 Hz, 2H), 6.57 (br s, 2H)
4-phenyl-pteridin-4-yl-amine
[0508] This compound was prepared by heating ammonium acetate with
the appropriate pyrazine in acetic acid for an hour. The product is
isolated by concentrating the solution by rotary evaporation and
washing with ether.
##STR00141##
[0509] .sup.1H NMR (500 MHz; DMSO-d.sub.6): .delta. 9.73 (s, 1H),
8.54 (s, 1H), 8.49 (dd, J=8.2 Hz, J=1.9 Hz, 2H), 8.46 (br s, 1H),
8.31 (br s, 1H), 7.60-7.55 (m, 3H)
Experimental Procedure
4-[2-(6-Phenyl-pteridin-4-ylamino)-ethyl]benzene-1,2-diol
##STR00142##
[0511] To a suspension of 3-hydroxytyramine hydrochloride (189.6
mg, 1.0 mmol) in 4 mL of glacial acetic acid was added
N'-(3-cyano-5-phenyl-pyrazin-2-yl)-N,N'-dimethyl-formamidine (251.3
mg, 1.0 mmol). The reaction was refluxed for 1.5 hours. The
progress of the reaction was monitored by LC/MS. After the reaction
had completed, the reaction mixture was cooled down to ambient
temperature and acetic acid was removed in vacuo. 5 mL of methanol
was added to the resulting residue and it was crushed with a
spatula into a fine suspension. 10 mL of 1:1 mixture of
acetonitrile/water was added to the suspension. The solid was
centrifuged down, washed with 20 mL of 1:1 mixture of
acetonitrile/water twice, 10 mL of methanol, 40 mL of diethyl ether
and dried in vacuo to give the product as a greenish-yellow solid.
58.5% yield. 96.9% purity by LC/MS (230 DAD). Mass-spec
[ES+]=360.5. .sup.1H NMR (DMSO-d6) 2.80-2.83 (m, 2H), 3.72-3.76 (m,
2H), 6.52-6.54 (dd, 1H), 6.65-6.67 (d, 1H), 6.68-6.69 (d, 1H),
7.56-7.61 (m, 3H), 8.45-8.47 (m, 2H), 8.63 (s, 1H), 8.68 (br.s,
1H), 8.80 (br.s, 1H, 8.91-8.94 (t, 1H), 9.72 (s, 1H). UV
.lamda..sub.max=239, 209, 279.
4-[(Phenyl-pteridin-4-ylamino)-methyl]-benzene-1,2-diol
##STR00143##
[0513] To a suspension of 3,4-dihydroxybenzylamine hydrobromide
(220.1 mg, 1.0 mmol) in 4 mL of glacial acetic acid was added
N'-(3-cyano-5-phenyl-pyrazin-2-yl)-N,N'-dimethyl-formamidine (251.3
mg, 1.0 mmol). The reaction was refluxed for 4 hours. The progress
of the reaction was monitored by LC/MS. After the reaction had
completed, the reaction mixture was cooled down to ambient
temperature and acetic acid was removed in vacuo. 5 mL of methanol
was added to the resulting residue and it was crushed with a
spatula into a fine suspension. The suspension was added to 45 mL
of diethyl ether. The solid was centrifuged down, washed with 45 mL
of diethyl ether twice and dried in vacuo to give the product as a
yellow solid. The product was purified by prep-HPLC, the major
product was collected and solvent was removed in vacuo. 99.6%
purity by LC/MS (230 DAD). Mass-spec [ES+]=346.5. .sup.1H NMR
(DMSO-d6) 5.56 (s, 2H), 6.68-6.70 (d, 1H), 6.75-6.77 (dd, 1H),
6.87-6.87 (d, 1H), 7.62-7.64 (m, 3H), 8.53-8.55 (m, 2H), 8.97 (s,
1H), 9.12 (s, 1H), 9.24 (s, 1H), 9.89 (s, 1H), 10.48 (br.s, 1H),
10.54 (br.s, 1H). UV .lamda..sub.max=245, 278, 210.
2,3-Bis(3,4-dihydroxyphenyl)-pyrido[2,3-b]pyrazin-6-ylamine
dihydrochloride salt
##STR00144##
[0515] 107.07 mg (1.0 mmol) of 2,3,6-triaminopyrimidine
dihydrochloride and 274.23 mg (1.0 mmol) of
3,3',4,4'-tetrahydroxybenzil were dissolved in 4 mL of 1:1 mixture
of dioxane-water. The reaction was refluxed for 8 hours. Then
solvent was removed in vacuo. The dark-yellow residue was dissolved
in 2 mL of methanol and this solution was added dropwise to 40 mL
of diethyl ether. The formed dark-yellow precipitate was collected,
washed with ether and dried in vacuo to give 370.0 mg (85% yield)
of the product. 100% purity by LC/MS (230 DAD). Mass-spec
[ES+]=363.8. 1H NMR (MeOH-d4) 6.70-6.75 (2H, dd), 6.81-6.92 (2H,
dd), 6.96-7.07 (2H, dd), 7.27 (1H, d), 8.34 (1H, d).
2,3-Bis(3-hydroxyphenyl)quinoxalin-6-ylamine dihydrochloride
salt
##STR00145##
[0517] 40.4 mg (0.206 mmol) of 1,2,4-benzenetriamine
dihydrochloride and 50 mg (0.20 mmol) of 3,3'-dihydroxybenzil were
dissolved in 2 mL of 1:1 mixture of dioxane-water. The reaction was
refluxed for 3 hours. Then solvent was removed in vacuo. The
residue was dissolved in 2 mL of methanol and this solution was
added dropwise to 40 mL of diethyl ether. The formed dark-red
precipitate was collected, washed with ether and dried in vacuo to
give 69.8 mg (92.6% yield) of the product. 97.6% purity by LC/MS
(230 DAD). Mass-spec [ES+]=330.8. .sup.1H NMR (500 MHz, MeOH-d4)
6.81-6.87 (2H, m), 6.96-6.98 (4H, m), 7.10 (1H, m), 7.13-7.16 (1H,
t), 7.28-7.31 (1H, t), 7.56-7.58 (1H, m), 8.04-8.06 (1H, d).
2,3-Bis(4-hydroxyphenyl)quinoxalin-6-ylamine dihydrochloride
salt
##STR00146##
[0519] 98.04 mg (0.5 mmol) of 1,2,4-benzenetriamine dihydrochloride
and 121.2 mg (0.5 mmol) of 4,4'-dihydroxybenzil were dissolved in 2
ml of 1:1 mixture of dioxane-water. The reaction was refluxed for 3
hours. Then solvent was removed in vacuo. The residue was dissolved
in 2 ml of methanol and this solution was added dropwise to 40 ml
of diethyl ether. The formed dark-red precipitate was collected,
washed with ether and dried in vacuo to give 168.3 mg (83.7% yield)
of the product. 98.7% purity by LC/MS (230 DAD). Mass-spec
[ES+]=330.8. .sup.1H NMR (500 MHz, MeOH-d4) 6.76-6.77 (2H, d),
6.87-6.89 (2H, d), 7.05-7.06 (1H, d), 7.29-7.31 (2H, d), 7.38-7.40
(2H, d), 7.50-7.52 (1H, m), 7.99-8.01 (1H, d).
2,3-Bis(3,4-dihydroxyphenyl)quinoxalin-6-ylamine dihydrochloride
salt
##STR00147##
[0521] 98.0 mg (0.5 mmol) of 1,2,4-benzenetriamine dihydrochloride
and 137.1 mg (0.5 mmol) of 3,3',4,4'-tetrahydroxybenzil were
dissolved in 3 ml of MeOH. The reaction was refluxed for 6 hours.
Then the reaction mixture was cooled to r.t. and added dropwise to
40 ml of diethyl ether. The formed dark-red precipitate was
collected, washed with ether and dried in vacuo to give 184.0 mg
(84.7% yield) of the product. 97.7% purity by LC/MS (230 DAD).
Mass-spec [ES+]=362.8. .sup.1H NMR (MeOH-d4) 6.73-6.75 (1H, d),
6.78-6.80 (1H, m), 6.88-6.89 (1H, m), 6.94-6.97 (3H, m), 7.03 (1H,
d), 7.49-7.51 (1H, dd), 7.97-7.99 (1H, d).
2-Hydroxy-5-(6-phenyl-pteridin-4-ylamino)-benzenesulfonic acid
##STR00148##
[0523] 70.1% yield. 83% purity by LC/MS (230 DAD). Mass-spec
[ES+]=396.8. .sup.1H NMR (DMSO-d6) 7.17-7.19 (1H, dd), 7.58-7.63
(3H, m), 7.80-7.82 (1H, dd), 7.993-7.999 (1H, d), 8.61-8.63 (2H,
m), 8.73 (1H, s), 9.80 (1H, s), 10.51-10.53 (3H, m).
5-(6-Phenyl-pteridin-4-ylamino)-quinolin-8-ol hydrochloride
salt
##STR00149##
[0525] 79.9% yield. 85% purity by LC/MS (230 DAD). Mass-spec
[ES+]=367.7. .sup.1H NMR (DMSO-d6) 7.39-7.40 (1H, m), 7.61-7.72
(3H, m), 7.73-7.77 (2H, m), 8.60-8.67 (4H, m), 9.01-9.02 (1H, m),
9.92 (1H, s), 11.58 (1H, br.s.)
[0526] General Procedure
##STR00150##
##STR00151##
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide
##STR00152##
[0528] 4-Bromo-2-nitro-phenylamine (2.48 g, 11.4 mmol) was mixed
with cynamide (1.51 g, 36 mmol) in a 20 mL vial. The mixture was
heated to 100.degree. C. till the mixture was totally melted. The
mixture was cooled down to room temperature and 6.5 ml concentrated
HCl was added. The mixture was heated at 100.degree. C. for 40
minutes and cool down in ice water. 6.5 ml 14M NaOH was carefully
added to the above reaction mixture. The resulted mixture was
heated at 100.degree. C. for 2 hours then cool down to room
temperature. After filtration, the precipitate was washed several
times with water, methanol and ditheylether to remove the starting
material. 0.739 g product was obtained. Yield: 27%. ESI-MS:
[M+H].sup.+, 241, 243; .sup.1H NMR (DMSO-d.sub.6): .delta. 7.48 (d,
J=9.02 Hz, 1H), 7.89 (dd, J.sub.1=9.02 Hz, J.sub.2=2.14 Hz, 1H),
8.26 (d, J=2.14 Hz, 1H).
7-Bromo-5-methyl-benzo[1,2,4]triazin-3-ylamine-1-oxide
##STR00153##
[0530] 4-Bromo-2-methyl-6-nitro-phenylamine (1 g, 4.33 mmol) was
mixed with cynamide (0.5 g, 12 mmol) and 5 g pyridine HCl in a 20
ml vial. The mixture was heated to reflux overnight. The mixture
was cooled down to room temperature and 10% NaOH was carefully
added. The resulted mixture was heated at 100.degree. C. for 2
hours then cool down to room temperature. After filtration, the
precipitate was washed several times with water, acetone and
ditheylether to remove the starting material. 0.4 g product was
obtained. Yield: 36%. ESI-MS: [M+H].sup.+, 255, 257; .sup.1H NMR
(DMSO-d.sub.6): .delta. 2.45 (s, 3H), 7.81 (d, J=1.97 Hz, 1H), 8.26
(d, J=1.97 Hz, 1H).
7-Benzo[1,3]dioxol-5-yl-benzo[1,2,4]triazin-3-ylamine-1-oxide
##STR00154##
[0532] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (50 mg, 0.21 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
3,4-(Methylenedioxy)phenylboronic acid (68.6 mg, 0.41 mmol)
dissolved in 1 ml ethanol and potassium carbonate (32.4 mg, 0.3
mmol) dissolved in 1 ml water were added. Triphenylphosphine (9 mg,
0.034 mmol) and tris(dibenzylideneacetone) dipalladium (0) (9 mg,
9.83 umol) were added to the mixture. The mixture was reflux
overnight. The crude product was poured into 50 ml saturated
NaHCO.sub.3 solution, and CH.sub.2Cl.sub.2 was used to extract the
product. Solvent in the organic phase was removed under vacuum. The
resulted residue was purified by preparative HPLC. 20 mg
7-Benzo[1,3]dioxol-5-yl-benzo[1,2,4]triazin-3-ylamine-1-oxide was
isolated. Yield: 34.5%; ESI-MS: [M+H].sup.+, 283; .sup.1H NMR
(DMSO-d.sub.6): .delta. 6.09 (s, 2H), 7.04 (d, J=8.12 Hz, 1H), 7.27
(dd, J.sub.1=7.88 Hz, J.sub.2=1.58 Hz, 1H), 7.37 (s, 1H), 7.58 (d,
J=8.12 Hz, 1H), 8.10 (dd, J=8.86 Hz, J.sub.2=1.86 Hz, 1H), 8.25 (d,
J=1.86 Hz, 1H).
7-Benzo[1,3]dioxol-5-yl-benzo[1,2,4]triazin-3-ylamine
##STR00155##
[0534] 10 mg
7-Benzo[1,3]dioxol-5-yl-benzo[1,2,4]triazin-3-ylamine-1-oxide was
dissolved in a mixture of 2 ml N,N-Dimethylacetamide and 1 ml ethyl
alcohol in a 20 ml vial with a septum. Catalytic amount of 10%
Palladium on carbon was added to the mixture. A balloon filled with
hydrogen was placed on the top of the vial. The mixture was stirred
at room temperature for 2 hours. Celite was used to remove the
palladium and carbon. Preparative HPLC was used to isolate the
final product. 5 mg
7-(2,6-Dimethyl-phenyl)-benzo[1,2,4]triazin-3-ylamine was obtained.
Yield: 53%; ESI-MS: [M+H].sup.+, 267; .sup.1H NMR (DMSO-d.sub.6):
.delta. 6.09 (s, 2H), 7.04 (d, J=8.00 Hz, 1H), 7.33 (dd,
J.sub.1=7.91 Hz, J.sub.2=1.76 Hz, 1H), 7.46 (d, J=1.51 Hz, 1H),
7.58 (d, J=8.84 Hz, 1H), 8.12 (dd, J.sub.1=8.84 Hz, J.sub.2=1.96
Hz, 1H), 8.39 (d, J=1.96 Hz, 1H).
7-(2,6-Dimethyl-phenyl)-benzo[1,2,4]triazin-3-ylamine
##STR00156##
[0536] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (100 mg, 0.42 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
2,6-dimethylphenylboronic acid (240 mg, 1.6 mmol) dissolved in 1 ml
ethanol and potassium carbonate (64 mg, 0.6 mmol) dissolved in 1 ml
water were added. Triphenylphosphine (9 mg, 0.034 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (9 mg, 9.83 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was poured into 50 ml saturated NaHCO.sub.3 solution, and
CH.sub.2Cl.sub.2 was used to extract the product. Solvent in the
organic phase was removed under vacuum. The residue was dissolved
in a mixture of 2 ml N,N-Dimethylacetamide and 1 ml ethyl alcohol
in a 20 ml vial with a septum. Catalytic amount of 10% Palladium on
carbon was added to the mixture. A balloon filled with hydrogen was
placed on the top of the vial. The mixture was stirred at room
temperature for 2 hours. Celite was used to remove the palladium
and carbon. Preparative HPLC was used to isolate the final product.
60 mg 7-(2,6-Dimethyl-phenyl)-benzo[1,2,4]triazin-3-ylamine was
obtained. Yield: 60%; ESI-MS: [M+H].sup.+, 251; .sup.1H NMR
(DMSO-d.sub.6): .delta. 2.03 (s, 6H), 7.23-7.16 (m, 3H), 7.62-7.58
(m, 2H), 7.95 (m, 1H).
7-(4-Phenoxy-phenyl)-benzo[1,2,4]triazin-3-ylamine
##STR00157##
[0538] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (100 mg, 0.42 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
4-Phenoxyphenylboronic acid (177 mg, 0.83 mmol) dissolved in 1 ml
ethanol and potassium carbonate (64 mg, 0.6 mmol) dissolved in 1 ml
water were added. Triphenylphosphine (9 mg, 0.034 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (9 mg, 9.83 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was poured into 50 ml saturated NaHCO.sub.3 solution, and
CH.sub.2Cl.sub.2 was used to extract the product. Solvent in the
organic phase was removed under vacuum. The residue was dissolved
in a mixture of 2 ml N,N-Dimethylacetamide and 1 ml ethyl alcohol
in a 20 ml vial with a septum. Catalytic amount of 10% Palladium on
carbon was added to the mixture. A balloon filled with hydrogen was
placed on the top of the vial. The mixture was stirred at room
temperature for 2 hours. Celite was used to remove the palladium
and carbon. Preparative HPLC was used to isolate the final product.
20 mg 3-(3-Amino-benzo[1,2,4]triazin-7-yl)-benzonitrile was
obtained. Yield: 15.4%; ESI-MS: [M+H].sup.+, 315; .sup.1H NMR
(DMSO-d.sub.6): .delta. 7.09-7.13 (m, 5H), 7.44 (m, 2H), 7.62 (d,
J=8.89 Hz, 2H), 7.87 (m, 2H), 8.15 (dd, J.sub.1=8.89 Hz,
J.sub.2=2.34 Hz, H), 8.43 (d, J=2.34 Hz, 1H).
7-(2,6-Dimethoxy-phenyl)-benzo[1,2,4]triazin-3-ylamine
##STR00158##
[0540] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (100 mg, 0.42 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
2,6-dimethoxy-phenylboronic acid (302 mg, 1.66 mmol) dissolved in 1
ml ethanol and potassium carbonate (64 mg, 0.6 mmol) dissolved in 1
ml water were added. Triphenylphosphine (9 mg, 0.034 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (9 mg, 9.83 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was poured into 50 ml saturated NaHCO.sub.3 solution, and
CH.sub.2Cl.sub.2 was used to extract the product. Solvent in the
organic phase was removed under vacuum. The residue was dissolved
in a mixture of 2 ml N,N-Dimethylacetamide and 1 ml ethyl alcohol
in a 20 ml vial with a septum. Catalytic amount of 10% Palladium on
carbon was added to the mixture. A balloon filled with hydrogen was
placed on the top of the vial. The mixture was stirred at room
temperature for 2 hours. Celite was used to remove the palladium
and carbon. Preparative HPLC was used to isolate the final product.
40 mg 7-(2,6-Dimethoxy-phenyl)-benzo[1,2,4]triazin-3-ylamine was
obtained. Yield: 34.2%, ESI-MS: [M+H].sup.+, 283; .sup.1H NMR
(DMSO-d.sub.6): .delta. 3.71 (s, 6H), 6.80 (d, J=8.47 Hz, 2H), 7.36
(t, J=8.39 Hz, 1H), 7.52 (d, J=8.85 Hz, 1H), 7.66 (dd, J.sub.1=8.85
Hz, J.sub.2=1.91 Hz, 1H), 8.00 (d, J=1.91 Hz, 1H).
7-(4-t-Butyl-phenyl)-benzo[1,2,4]triazin-3-ylamine
##STR00159##
[0542] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (100 mg, 0.42 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
4-t-butyl-phenylboronic acid (148 mg, 0.83 mmol) dissolved in 1 ml
ethanol and potassium carbonate (64 mg, 0.6 mmol) dissolved in 1 ml
water were added. Triphenylphosphine (9 mg, 0.034 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (9 mg, 9.83 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was poured into 50 ml saturated NaHCO.sub.3 solution, and
CH.sub.2Cl.sub.2 was used to extract the product. Solvent in the
organic phase was removed under vacuum. The residue was dissolved
in a mixture of 2 ml N,N-Dimethylacetamide and 1 ml ethyl alcohol
in a 20 ml vial with a septum. Catalytic amount of 10% Palladium on
carbon was added to the mixture. A balloon filled with hydrogen was
placed on the top of the vial. The mixture was stirred at room
temperature for 2 hours. Celite was used to remove the palladium
and carbon. Preparative HPLC was used to isolate the final product.
20 mg 7-(4-t-Butyl-phenyl)-benzo[1,2,4]triazin-3-ylamine was
obtained. Yield: 18%, ESI-MS: [M+H].sup.+, 279; .sup.1H NMR
(DMSO-d.sub.6): .delta. 1.34 (s, 9H), 7.53 (d, J=8.66 Hz, 2H), 7.61
(d, J=8.85 Hz, 1H), 7.77 (d, J=8.66 Hz, 2H), 8.16 (dd, J.sub.1=8.84
Hz, J.sub.2=1.89 Hz, 1H), 8.43 (d, J=1.89 Hz, 1H).
7-(2-Trifluoromethyl-phenyl)-benzo[1,2,4]triazin-3-ylamine
##STR00160##
[0544] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (100 mg, 0.42 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
2-trifluoromethyl phenylboronic acid (157 mg, 0.83 mmol) dissolved
in 1 ml ethanol and potassium carbonate (64 mg, 0.6 mmol) dissolved
in 1 ml water were added. Triphenylphosphine (9 mg, 0.034 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (9 mg, 9.83 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was poured into 50 ml saturated NaHCO.sub.3 solution, and
CH.sub.2Cl.sub.2 was used to extract the product. Solvent in the
organic phase was removed under vacuum. The residue was dissolved
in a mixture of 2 ml N,N-Dimethylacetamide and 1 ml ethyl alcohol
in a 20 ml vial with a septum. Catalytic amount of 10% Palladium on
carbon was added to the mixture. A balloon filled with hydrogen was
placed on the top of the vial. The mixture was stirred at room
temperature for 2 hours. Celite was used to remove the palladium
and carbon. Preparative HPLC was used to isolate the final product.
20 mg 7-(2-Trifluoromethyl-phenyl)-benzo[1,2,4]triazin-3-ylamine
was obtained. Yield: 16.5%, ESI-MS: [M+H].sup.+, 291; .sup.1H NMR
(DMSO-d.sub.6): .delta. 7.56 (d, J=7.56 Hz, 1H), 7.60 (d, J=8.66
Hz, 1H), 7.68-7.80 (m, 3H), 7.89 (d, J=7.56 Hz, 1H), 8.11 (d,
J=1.46 Hz, 1 .mu.l).
7-Biphenyl-4-yl-benzo[1,2,4]triazin-3-ylamine
##STR00161##
[0546] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (100 mg, 0.42 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
4-biphenylboronic acid (164 mg, 0.83 mmol) dissolved in 1 ml
ethanol and potassium carbonate (64 mg, 0.6 mmol) dissolved in 1 ml
water were added. Triphenylphosphine (9 mg, 0.034 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (9 mg, 9.83 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was poured into 50 ml saturated NaHCO.sub.3 solution, and
CH.sub.2Cl.sub.2 was used to extract the product. Solvent in the
organic phase was removed under vacuum. The residue was dissolved
in a mixture of 2 ml N,N-Dimethylacetamide and 1 ml ethyl alcohol
in a 20 ml vial with a septum. Catalytic amount of 10% Palladium on
carbon was added to the mixture. A balloon filled with hydrogen was
placed on the top of the vial. The mixture was stirred at room
temperature for 2 hours. Celite was used to remove the palladium
and carbon. Preparative HPLC was used to isolate the final product.
15 mg 7-Biphenyl-4-yl-benzo[1,2,4]triazin-3-ylamine was obtained.
Yield: 12.1%, ESI-MS: [M+H].sup.+, 299; .sup.1H NMR (DMSO-d.sub.6):
.delta. 7.41 (m, 1H), 7.50 (m, 2H), 7.55 (m, 2H), 7.64 (d, J=8.84
Hz, 1H), 7.83 (m, 2H), 7.96 (m, 2H), 8.24 (dd, J.sub.1=8.84 Hz,
J.sub.2=1.93 Hz, 1H), 8.53 (d, J=1.93 Hz, 1H).
7-Benzofuran-2-yl-benzo[1,2,4]triazin-3-ylamine
##STR00162##
[0548] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (100 mg, 0.42 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
2-Benzofuranboronic acid (134 mg, 0.83 mmol) dissolved in 1 ml
ethanol and potassium carbonate (64 mg, 0.6 mmol) dissolved in 1 ml
water were added. Triphenylphosphine (9 mg, 0.034 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (9 mg, 9.83 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was poured into 50 ml saturated NaHCO.sub.3 solution, and
CH.sub.2Cl.sub.2 was used to extract the product. Solvent in the
organic phase was removed under vacuum. The residue was dissolved
in a mixture of 2 ml N,N-Dimethylacetamide and 1 ml ethyl alcohol
in a 20 ml vial with a septum. Catalytic amount of 10% Palladium on
carbon was added to the mixture. A balloon filled with hydrogen was
placed on the top of the vial. The mixture was stirred at room
temperature for 2 hours. Celite was used to remove the palladium
and carbon. Preparative HPLC was used to isolate the final product.
10 mg 7-Benzofuran-2-yl-benzo[1,2,4]triazin-3-ylamine was obtained.
Yield: 9.3%, ESI-MS: [M+H].sup.+, 263; .sup.1H NMR (DMSO-d.sub.6):
.delta. 6.54 (s, 1H), 7.29 (t, J=7.22 Hz, 1H), 7.36 (t, J=7.23 Hz,
1H), 7.64-7.71 (m, 3H), 7.34 (dd, J.sub.1=8.86 Hz, J.sub.2=1.86 Hz,
1H), 8.63 (d, J=1.86 Hz, 1H).
7-Dibenzofuran-4-yl-benzo[1,2,4]triazin-3-ylamine
##STR00163##
[0550] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (100 mg, 0.42 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
4-Dibenzofuranboronic acid (176 mg, 0.83 mmol) dissolved in 1 ml
ethanol and potassium carbonate (64 mg, 0.6 mmol) dissolved in 1 ml
water were added. Triphenylphosphine (9 mg, 0.034 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (9 mg, 9.83 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was poured into 50 ml saturated NaHCO.sub.3 solution, and
CH.sub.2Cl.sub.2 was used to extract the product. Solvent in the
organic phase was removed under vacuum. The residue was dissolved
in a mixture of 2 ml N,N-Dimethylacetamide and 1 ml ethyl alcohol
in a 20 ml vial with a septum. Catalytic amount of 10% Palladium on
carbon was added to the mixture. A balloon filled with hydrogen was
placed on the top of the vial. The mixture was stirred at room
temperature for 2 hours. Celite was used to remove the palladium
and carbon. Preparative HPLC was used to isolate the final product.
5 mg 7-Dibenzofuran-4-yl-benzo[1,2,4]triazin-3-ylamine was
obtained. Yield: 3.9%, ESI-MS: [M+H].sup.+, 263; .sup.1H NMR
(DMSO-d.sub.6): .delta. 7.46 (t, J=7.62 Hz, 1H), 7.57 (t, J=7.92
Hz, 2H), 7.72 (t, J=8.85 Hz, 1H), 7.80 (d, J=8.20 Hz, 1H), 7.90 (d,
J=8.07 Hz, 1H), 8.23 (m, 2H), 8.38 (dd, J=8.84 Hz, J.sub.2=2.06 Hz,
1H), 8.63 (d, J=2.06 Hz, 1H).
7-Naphthalen-1-yl-benzo[1,2,4]triazin-3-ylamine
##STR00164##
[0552] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (100 mg, 0.42 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
1-Naphthylboronic acid (143 mg, 0.83 mmol) dissolved in 1 ml
ethanol and potassium carbonate (64 mg, 0.6 mmol) dissolved in 1 ml
water were added. Triphenylphosphine (9 mg, 0.034 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (9 mg, 9.83 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was poured into 50 ml saturated NaHCO.sub.3 solution, and
CH.sub.2Cl.sub.2 was used to extract the product. Solvent in the
organic phase was removed under vacuum. The residue was dissolved
in a mixture of 2 ml N,N-Dimethylacetamide and 1 ml ethyl alcohol
in a 20 ml vial with a septum. Catalytic amount of 10% Palladium on
carbon was added to the mixture. A balloon filled with hydrogen was
placed on the top of the vial. The mixture was stirred at room
temperature for 2 hours. Celite was used to remove the palladium
and carbon. Preparative HPLC was used to isolate the final product.
10 mg 7-Naphthalen-1-yl-benzo[1,2,4]triazin-3-ylamine was obtained.
Yield: 8.8%, ESI-MS: [M+H].sup.+, 273; .sup.1H NMR (DMSO-d.sub.6):
.delta. 7.54-7.69 (m, 5H), 7.84 (d, J=8.31 Hz, 1H), 7.94 (dd,
J.sub.1=8.60 Hz, J.sub.2=1.68 Hz, 1H), 8.05 (m, 2H), 8.26 (d,
J=1.68 Hz, 1H).
3-(3-Amino-benzo[1,2,4]triazin-7-yl)-phenol
##STR00165##
[0554] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (100 mg, 0.42 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
3-hydroxyphenylboronic acid (114.5 mg, 0.83 mmol) dissolved in 1 ml
ethanol and potassium carbonate (64 mg, 0.6 mmol) dissolved in 1 ml
water were added. Triphenylphosphine (9 mg, 0.034 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (9 mg, 9.83 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was poured into 50 ml saturated NaHCO.sub.3 solution, and
CH.sub.2Cl.sub.2 was used to extract the product. Solvent in the
organic phase was removed under vacuum. The residue was dissolved
in a mixture of 2 ml N,N-Dimethylacetamide and 1 ml ethyl alcohol
in a 20 ml vial with a septum. Catalytic amount of 10% Palladium on
carbon was added to the mixture. A balloon filled with hydrogen was
placed on the top of the vial. The mixture was stirred at room
temperature for 2 hours. Celite was used to remove the palladium
and carbon. Preparative HPLC was used to isolate the final product.
15 mg 3-(3-Amino-benzo[1,2,4]triazin-7-yl)-phenol was obtained.
Yield: 15%, ESI-MS: [M+H].sup.+, 239; .sup.1H NMR (DMSO-d.sub.6):
.delta. 6.82 (dd, J.sub.1=7.94 Hz, J.sub.2=1.98 Hz, 1H), 7.17 (m,
1H), 7.23 (d, J=7.80 Hz, 1H), 7.31 (t, J=7.73 Hz, 1H), 7.60 (d,
J=8.83 Hz, 1H), 8.08 (dd, J.sub.1=8.83 Hz, J.sub.2=1.94 Hz, 1H),
8.36 (d, J=1.94 Hz, 1H).
[7-(2,6-Dimethyl-phenyl)-benzo[1,2,4]triazin-3-yl]-phenyl-amine
##STR00166##
[0555] 7-(2,6-Dimethyl-phenyl)-benzo[1,2,4]triazin-3-ylamine (24
mg, 0.096 mmol) was dissolved in aniline, sulfamic acid (18 mg,
0.19 mmol) was added. The mixture was reflux overnight. The final
product was isolated by preparative HPLC. Yield: 32%. ESI-MS:
[M+H].sup.+, 327; .sup.1H NMR (DMSO-d.sub.6): .delta. 2.05 (s, 6H),
7.09 (t, J=7.35 Hz, 1H), 7.18-7.25 (m, 3H), 7.40 (m, 2H), 7.71 (dd,
J.sub.1=8.5 Hz, J.sub.2=1.9 Hz, 1H), 7.84 (d, J=8.5 Hz, 1H), 8.00
(d, J=7.6 Hz, 2H), 8.11 (d, J=1.9 Hz, 1H).
(7-Bromo-5-methyl-benzo[1,2,4]triazin-3-yl)-phenyl-amine
##STR00167##
[0556] 7-Bromo-5-methyl-benzo[1,2,4]triazin-3-ylamine-1-oxide (266
mg, 1.04 mmol) was dissolved in 5 ml acetic acid in a 20 ml vial, a
few drops of water was added followed by adding of 100 mg Fe
powder. The mixture was kept at 100.degree. C. for 30 minutes. The
solvent was removed under vacuum. The residue was dissolved in 5 ml
aniline, sulfamic acid (202 mg, 2.08 mmol) was added to the
mixture. The mixture was heat at 140.degree. C. for overnight. The
final product was isolated by preparative HPLC. Yield: 18.3%,
ESI-MS: [M+H].sup.+, 315, 317.
(7-Bromo-5-methyl-benzo[1,2,4]triazin-3-yl)-[3-(4-methyl-piperazin-1-yl)-p-
ropyl]-amine
##STR00168##
[0558] 7-Bromo-5-methyl-benzo[1,2,4]triazin-3-ylamine-1-oxide (200
mg, 0.78 mmol) was dissolved in 5 ml acetic acid in a 20 ml vial, a
few drops of water was added followed by adding of 100 mg Fe
powder. The mixture was kept at 100.degree. C. for 30 minutes. The
solvent was removed under vacuum. The residue was dissolved in 5 ml
3-(4-Methyl-piperazin-1-yl)-propylamine, sulfamic acid (152 mg,
1.57 mmol) was added to the mixture. The mixture was heat at
140.degree. C. for overnight. The final product was isolated by
preparative HPLC. Yield: 67.3%, ESI-MS: [M+H].sup.+, 379, 381.
.sup.1H NMR (DMSO-d.sub.6): .delta. 1.05 (m, 2H), 1.97 (s, 2H),
2.77-3.20 (b, 8H), 3.5 (b, 8H), 7.84 (d, J=1.96 Hz, 1H), 8.29 (d,
J=1.96 Hz, 1H).
[5-Methyl-7-(2,4,6-trimethyl-phenyl)-benzo[1,2,4]triazin-3-yl]-phenyl-amin-
e
##STR00169##
[0560] To a solution of
(7-Bromo-5-methyl-benzo[1,2,4]triazin-3-yl)-phenyl-amine (10 mg,
0.032 mmol) dissolved in 2 ml N,N-Dimethylacetamide in a 20 ml
vial, 2,4,6-trimethylphenylboronic acid (21 mg, 0.128 umol)
dissolved in 1 ml ethanol and potassium carbonate (6.4 mg, 0.06
mmol) dissolved in 1 ml water were added. Triphenylphosphine (1 mg,
0.0038 mmol) and tris(dibenzylideneacetone) dipalladium (0) (1 mg,
1.09 umol) were added to the mixture. The mixture was reflux
overnight. The crude product was filtered and purified by
preparative HPLC. 3 mg
[5-Methyl-7-(2,4,6-trimethyl-phenyl)-benzo[1,2,4]triazin-3-yl]-phenyl-ami-
ne was isolated. Yield: 26.8%; ESI-MS: [M+H].sup.+, 355; .sup.1H
NMR (CDCl.sub.3): .delta. 2.06 (s, 6H), 2.36 (s, 3H), 2.72 (s, 3H),
6.99 (s, 2H), 7.17 (m, 1H), 7.45 (m, 2H), 7.57 (m, 1H), 7.89 (d,
J=1.36 Hz, 1H), 7.94 (d, J=8.76 Hz, 2H).
[7-(2-Fluoro-6-methoxy-phenyl)-5-methyl-benzo[1,2,4]triazin-3-yl]-phenyl-a-
mine
##STR00170##
[0562] To a solution of
(7-Bromo-5-methyl-benzo[1,2,4]triazin-3-yl)-phenyl-amine (10 mg,
0.032 mmol) dissolved in 2 ml N,N-Dimethylacetamide in a 20 ml
vial, 2-Fluoro-6-methoxy-phenylboronic acid (22 mg, 0.128 mmol)
dissolved in 1 ml ethanol and potassium carbonate (6.4 mg, 0.06
mmol) dissolved in 1 ml water were added. Triphenylphosphine (1 mg,
0.0038 mmol) and tris(dibenzylideneacetone) dipalladium (0) (1 mg,
1.09 umol) were added to the mixture. The mixture was reflux
overnight. The crude product was filtered and purified by
preparative HPLC. 2 mg
[7-(2-Fluoro-6-methoxy-phenyl)-5-methyl-benzo[1,2,4]triazin-3-yl]-phenyl--
amine was isolated. Yield: 17.5%; ESI-MS: [M+H].sup.+, 361; .sup.1H
NMR (CDCl.sub.3): .delta. 2.73 (s, 3H), 3.83 (s, 3H), 6.83-6.86 (m,
2H), 7.14 (m, 1H), 7.34 (m, 1H), 7.45 (m, 2H), 7.75 (s, 1H), 7.92
(m, 2H), 8.24 (s, 1H).
[7-(2,6-Dimethoxy-phenyl)-5-methyl-benzo[1,2,4]triazin-3-yl]-phenyl-amine
##STR00171##
[0564] To a solution of
(7-Bromo-5-methyl-benzo[1,2,4]triazin-3-yl)-phenyl-amine (10 mg,
0.032 mmol) dissolved in 2 ml N,N-Dimethylacetamide in a 20 ml
vial, 2,6-dimethoxy-phenylboronic acid (23 mg, 0.126 mmol)
dissolved in 1 ml ethanol and potassium carbonate (6.4 mg, 0.06
mmol) dissolved in 1 ml water were added. Triphenylphosphine (1 mg,
0.0038 mmol) and tris(dibenzylideneacetone) dipalladium (0) (1 mg,
1.09 umol) were added to the mixture. The mixture was reflux
overnight. The crude product was filtered and purified by
preparative HPLC. 5 mg
[7-(2,6-Dimethoxy-phenyl)-5-methyl-benzo[1,2,4]triazin-3-yl]-phenyl-amine
was isolated. Yield: 42.4%; ESI-MS: [M+H].sup.+, 373; .sup.1H NMR
(CDCl.sub.3): .delta. 2.72 (s, 3H), 3.78 (s, 6H), 6.70 (d, J=8.4
Hz, 2H), 7.13 (m, 1H), 7.35 (t, J=8.38 Hz, 1H), 7.44 (m, 2H), 7.89
(m, 1H), 7.92 (dd, J.sub.1=8.78 Hz, J.sub.2=2.02 Hz, 2H), 8.18 (d,
J=2.02 Hz, 1H).
[7-(2,6-Dimethyl-phenyl)-5-methyl-benzo[1,2,4]triazin-3-yl]-phenyl-amine
##STR00172##
[0566] To a solution of
(7-Bromo-5-methyl-benzo[1,2,4]triazin-3-yl)-phenyl-amine (60 mg,
0.19 mmol) dissolved in 3 ml N,N-Dimethylacetamide in a 20 ml vial,
2,6-dimethyl-phenylboronic acid (114 mg, 0.76 mmol) dissolved in 2
ml ethanol and potassium carbonate (31 mg, 0.3 mmol) dissolved in 1
ml water were added. Triphenylphosphine (4.5 mg, 0.0171 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (4.5 mg, 4.9 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was filtered and purified by preparative HPLC. 30 mg
[7-(2,6-Dimethyl-phenyl)-5-methyl-benzo[1,2,4]triazin-3-yl]-phenyl-amine
was isolated. Yield: 46%; ESI-MS: [M+H].sup.+, 341; .sup.1H NMR
(DMSO-d6): .delta. 2.05 (s, 6H), 2.67 (s, 3H), 7.07 (t, J=7.33 Hz,
1H), 7.17-7.24 (m, 3H), 7.41 (t, J=7.56 Hz, 2H), 7.62 (d, J=1.49
Hz, 1H), 7.93 (d, J=1.49 Hz, 1H), 8.05 (d, J=7.72 Hz, 1H).
7-Naphthalen-2-yl-benzo[1,2,4]triazin-3-ylamine-1-oxide
##STR00173##
[0568] To a solution of
7-Bromo-benzo[1,2,4]triazin-3-ylamine-1-oxide (100 mg, 0.42 mmol)
dissolved in 6 ml N,N-Dimethylacetamide in a 20 ml vial,
2-Naphthylboronic acid (143 mg, 0.83 mmol) dissolved in 1 ml
ethanol and potassium carbonate (64 mg, 0.6 mmol) dissolved in 1 ml
water were added. Triphenylphosphine (9 mg, 0.034 mmol) and
tris(dibenzylideneacetone) dipalladium (0) (9 mg, 9.83 umol) were
added to the mixture. The mixture was reflux overnight. The crude
product was poured into 50 ml saturated NaHCO.sub.3 solution, and
CH.sub.2Cl.sub.2 was used to extract the product. Solvent in the
organic phase was removed under vacuum. Preparative HPLC was used
to isolate the final product. 20 mg
7-Naphthalen-2-yl-benzo[1,2,4]triazin-3-ylamine-1-oxide was
obtained. Yield: 16.7%, ESI-MS: [M+H].sup.+, 289; .sup.1H NMR
(DMSO-d6): .delta. 7.56 (m, 2H), 7.68 (d, J=8.84 Hz, 1H), 7.95 (m,
2H), 8.05 (d, J=8.64 Hz, 2H), 8.33 (dd, J.sub.1=8.84 Hz,
J.sub.2=1.87 Hz, 1H), 8.38 (s, 1H), 8.51 (d, J=1.87 Hz, 1H).
[0569] General Procedure for the 6-Alkyl Substituted Pteridine
Synthesis
##STR00174##
6-Bromomethyl-2,4-pteridinediamine
##STR00175##
[0571] To the solution of dibromotriphenylphosphine (2.4337 g, 5.76
mmol) of 2 ml anhydrous N,N-dimethylacetamide was added
(2,4-Diamino-Pteridin-6-yl)-methanol hydrobromide (335.8 mg, 1.747
mmol). The mixture is the stirred at RT for overnight. The solution
was treated with benzene. The filtered solid was then successively
treated with benzene and ether and evaporate the remaining solid.
The residue was dissolved in minimum 48% HBr at RT which then was
added MeCN to give a tan solid precipitate. Collect the solid in
ice water bath and wash it with MeCN and ether. 352 mg product was
obtained. Yield 60%; .sup.1H NMR (500 MHz, DMSO-d6): .delta.
4.86021 (s, 2H), 9.01 (s, 1H), 9.15 (s, 2H), 9.22 (s, 2H); ESI-MS:
255, 257 (M.sup.++1)
2-[(2,4-Diamino-pteridin-6-ylmethyl)-amino]-3-(4-hydroxy-phenyl)-propionic
acid tert-butyl ester
##STR00176##
[0573] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (31.2 mg, 0.116 mmol) in anhydrous N,N
dimethylacetamide was added 2-amino-3-(4-hydroxy-phenyl)-propionic
acid tert-butyl ester (30.22 mg, 0.127 mmol). The reaction mixture
was stirred at 50.degree. C. overnight. The crude product was
poured into saturated bicarbonate solution. The resulted
precipitate was collected and purified by preparative HPLC. 17.2 mg
product was obtained. Yield: 71%; .sup.1H NMR (500 MHz, DMSO-d6):
.delta. 1.33577 (s, 9H), 2.94185-3.02295 (m, 2H), 3.6550 (b, 1H),
4.0878 (s, 2H), 6.70174-6.72384 (dd, 8.545 Hz, J.sub.2=2.59 Hz,
2H), 7.02394-7.04103 (d, J=8.545 Hz, 2H); 9.38501 (s, 1H); ESI-MS:
412 (M.sup.++1)
6-[{(Pyridin-2-ylmethyl)-amino]-methyl}-2,4-pteridinediamine
##STR00177##
[0575] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (51 mg, 0.2 mmol) in anhydrous N,N dimethylacetamide
was added 2-(aminomethyl)pyridine (22.48 ul, 0.22 mmol). The
reaction mixture was stirred at 50.degree. C. overnight. The crude
product was poured into saturated bicarbonate solution. The
resulted precipitate was collected and purified by preparative
HPLC. 32.3 mg product was obtained. Yield: 57%; .sup.1H NMR (500
MHz, DMSO-d6): .delta. 3.93801 (s, 2H), 4.05772 (s, 2H),
7.5758-7.6003 (m, 1H), 7.97993-8.00181 (m, 1H), 8.49332-8.50942 (d,
J=8.05 Hz, 1H), 8.62592-8.64301 (d, J=8.545 Hz, 1H), 8.9938 (s,
1H); ESI-MS: 283 (M.sup.++1)
6-{[(Naphthalen-1-yl-methyl)-amino]-methyl}-2,4-pteridinediamine
##STR00178##
[0577] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (51 mg, 0.2 mmol) in anhydrous N,N dimethylacetamide
was added 1-aminomethyl-naphthalene (31.67 ul, 0.22 mmol). The
reaction mixture was stirred at 50.degree. C. overnight. The crude
product was poured into saturated bicarbonate solution. The
resulted precipitate was collected and purified by preparative
HPLC. 9 mg product was obtained. Yield: 15%; .sup.1H NMR (500 MHz,
DMSO-d6): .delta. 4.6479 (s, 2H), 4.7893 (s, 2H), 7.575-7.6244 (m,
3H), 7.74232-7.7570 (d, J=6.91 Hz, 1H), 7.9935-8.0276 (dd,
J.sub.1=8.06 Hz, J.sub.2=8.995 Hz, 2H), 8.1670-8.1831 (d, J=8.04
Hz, 1H), 8.8430 (s, 1H); ESI-MS: m/z 332 (M.sup.++1)
6-(Benzylamino-methyl)-2,4-pteridinediamine
##STR00179##
[0579] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (35.7 mg, 0.106 mmol) in anhydrous N,N
dimethylacetamide was added benzylamine (28.6 ul, 0.212 mmol). The
reaction mixture was stirred at 50.degree. C. overnight. The crude
product was poured into saturated bicarbonate solution. The
resulted precipitate was collected and purified by preparative
HPLC. 17.7 mg product was obtained. Yield: 62%; .sup.1H NMR (500
MHz, DMSO-d6): .delta. 4.30499 (s, 2H), 4.51599 (s, 2H),
7.42787-7.47298 (m, 3H), 7.50007-7.51927 (m, 2H), 8.87751 (s, 1H);
ESI-MS: m/z 282 (M.sup.++1)
6-{[(Adamantan-1-yl-methyl)-amino]-methyl}-2,4-pteridinediamine
##STR00180##
[0581] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (41.6 mg, 0.124 mmol) in anhydrous N,N
dimethylacetamide was added 1-aminomethyl adamantane (35.43 ul, 0.2
mmol). The reaction mixture was stirred at 50.degree. C. overnight.
The crude product was poured into saturated bicarbonate solution.
The resulted precipitate was collected and purified by preparative
HPLC. 12.7 mg product was obtained. Yield: 40%; .sup.1H NMR (500
MHz, DMSO-d6): .delta. 1.56754-1.67101 (m, 13H), 1.96741 (s, 2H),
2.71139 (s, 2H), 4.49166 (s, 2H), 8.89918 (s, 1H); ESI-MS: m/z 340
(M.sup.++1)
6-(3,4-Dimethoxy-benzylamino)-2,4-pteridinediamine
##STR00181##
[0583] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (59 mg, 0.176 mmol) in anhydrous N,N dimethylacetamide
was added 3,4-dimethoxy -benzylamine (51.15 ul, 0.3512 mmol). The
reaction mixture was stirred at 50.degree. C. overnight. The crude
product was poured into saturated bicarbonate solution. The
resulted precipitate was collected and purified by preparative
HPLC. 20.3 mg product was obtained. Yield: 34%; .sup.1H NMR (500
MHz, DMSO-d6): .delta. 3.67534 (s, 3H), 3.70494 (s, 3H), 4.05412
(b, 4H), 6.78852-6.80460 (d, J=8.04 Hz, 1H), 6.83624 (s, 1H),
6.83624-6.85393 (d, J=8.195 Hz, 1H); 8.96623 (s, 1H), 9.00584 (s,
2H), 9.5577 (s, 2H); ESI-MS: 342 (M.sup.++1)
6-[2,2-Dimethyl-propylamino)-methyl]-2,4-pteridinediamine
##STR00182##
[0585] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (75.2 mg, 0.2237 mmol) in anhydrous N,N
dimethylacetamide was added 2,2-dimethyl-propylamine (136.48 ul,
1.16 mmol). The reaction mixture was stirred at room temperature
overnight. The resulted precipitate was collected and purified by
preparative HPLC. 8.3 mg product was obtained. Yield: 14.2%;
.sup.1H NMR (500 MHz, DMSO-d6): .delta. 0.98591 (s, 9H), 2.82895
(s, 2H), 4.38765 (s, 2H), 8.77458 (s, 1H); ESI-MS: m/z 262
(M.sup.++1)
6-{[2-(3,4-Dimethoxy-phenyl)ethylamino]-methyl}-2,4-pteridinediamine
##STR00183##
[0587] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (55 mg, 0.1638 mmol) in anhydrous N,N
dimethylacetamide was added 2-(3,4-dimethoxyphenyl)ethylamine
hydrochloride (55 ul, 0.32 mmol). The reaction mixture was stirred
at 50.degree. C. overnight. The crude product was poured into
saturated bicarbonate solution. The resulted precipitate was
collected and purified by preparative HPLC. 3.8 mg product was
obtained. Yield: 19.6%; .sup.1H NMR (500 MHz, DMSO-d6): .delta.
2.75943-2.79062 (t, J=7.37 Hz, 2H), 2.92110-2.95356 (t, J=7.365 Hz,
2H), 3.72197 (s, 3H), 3.75135 (s, 3H), 4.54559 (s, 2H),
6.74441-6.77765 (dd, J.sub.1=8.26 Hz, J.sub.2=1.955 Hz, 1H),
6.84994 (s, 1H), 6.88406-6.90401 (dd, J=8.195 Hz, J.sub.2=1.735 Hz,
1H); 8.87126 (s, 1H); ESI-MS: m/z 356 (M.sup.++1)
6-{[2-(3,4-Dihydroxy-phenyl)ethylamino]-methyl}-2,4-pteridinediamine
##STR00184##
[0589] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (67.3 mg, 0.2003 mmol) in anhydrous N,N
dimethylacetamide was added 2-(3,4-dihydroxyphenyl)ethylamine (43.6
mg, 0.23 mmol). Under positive pressure of Argon, iPr.sub.2EtN
(32.63 ul) was added. The reaction mixture was stirred at
50.degree. C. for 4 hrs and then at Room temperature overnight. The
crude product was poured into saturated bicarbonate solution. The
resulted precipitate was collected and purified by preparative
HPLC. 14.8 mg product was obtained. Yield: 22.6%; .sup.1H NMR (500
MHz, DMSO-d6): .delta. 2.69242 (b, 4H), 4.03353 (s, 2H),
6.37542-6.39065 (d, J=7.615 Hz, 1H), 6.4851 (s, 1H),
6.56632-6.58226 (d, J=7.97 Hz, 1H), 8.80972 (s, 1H); ESI-MS: m/z
328 (M.sup.++1)
4-{2-[Di(2,4-diaminopteridin-6-yl-methyl)-amino]-ethyl}-benzene-1,2-diol
##STR00185##
[0591] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (67.3 mg, 0.2003 mmol) in anhydrous N,N
dimethylacetamide was added 2-(3,4-dihydroxyphenyl)ethylamine
hydrochloride (43.6 mg, 0.23 mmol). Under positive pressure of
Argon, iPr.sub.2EtN (32.63 ul) was added. The reaction mixture was
stirred at 50.degree. C. for 4 hrs and then at Room temperature
overnight. The crude product was poured into saturated bicarbonate
solution. The resulted precipitate was collected and purified by
preparative HPLC. 3.2 mg product was obtained. Yield: 6.4%; .sup.1H
NMR (500 MHz, DMSO-d6): .delta. 2.63154-2.63891 (m, 2H), 2.72839
(m, 2H), 4.03844 (s, 4H), 6.32227-6.33832 (d, J=8.025 Hz, 1H),
6.38857 (s, 1H), 6.51654-6.53241 (d, J=8.835 Hz, 1H), 8.67743 (s,
2H); ESI-MS: m/z 502 (M.sup.++1)
6-{[2-(3,4-Dihydroxy)-benzylamino]-methyl}-2,4-pteridinediamine
##STR00186##
[0593] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (64 mg, 0.1905 mmol) in anhydrous N,N
dimethylacetamide was added 2-(3,4-dihydroxybenzyl) amine
hydrochloride (36.795 mg, 0.23 mmol). Under positive pressure of
Argon, iPr.sub.2EtN (40.15 ul) was added. The reaction mixture was
stirred at 50.degree. C. for 4 hrs and then at Room temperature
overnight. The crude product was poured into saturated bicarbonate
solution. The resulted precipitate was collected and purified by
preparative HPLC. 7.8 mg product was obtained. Yield: 13.1%;
.sup.1H NMR (500 MHz, DMSO-d6): .delta. 3.91255 (s, 2H), 4.61898
(s, 2H), 6.6094-6.62572 (d, J=8.16 Hz, 1H), 6.64921-6.66517 (d,
J=7.98 Hz, 1H), 6.79669-6.79963 (d, J=1.47 Hz, 1H), 8.88104 (s,
1H); ESI-MS: 314 (M.sup.++1)
3-(4-tert-Butoxy-phenyl)-2-[(2,4-diamino-pteridin-6-ylmethyl)-amino]-propi-
onic acid tert-butyl ester
##STR00187##
[0595] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (53.7 mg, 0.1598 mmol) in anhydrous N,N
dimethylacetamide was added
2-amino-3-(4-tert-butoxy-phenyl)-propionic acid tert-butyl ester
hydrochloride (51.58 mg, 0.1758 mmol). Under positive pressure of
Argon, iPr.sub.2EtN (33.69 ul) was added. The reaction mixture was
stirred at 50.degree. C. for 4 hrs and then at room temperature
overnight. The crude product was poured into saturated bicarbonate
solution. The resulted precipitate was collected and purified by
preparative HPLC. 27.6 mg product was obtained. Yield: 41%; NMR
(500 MHz, DMSO-d6): .delta. 1.22491 (s, 9H), 1.26835 (s, 9H),
2.921-2.971 (m, 2H), 4.130 (b, 1H), 4.427 (s, 2H), 6.91485-6.93165
(d, J=8.4 Hz, 2H), 7.16037-7.17723 (d, J=8.43 Hz, 2H), 8.89353 (s,
1H); 9.13119 (s, 2H), 9.30829 (s, 2H); ESI-MS: m/z 468
(M.sup.+1)
1-{[di-(2,4-Diaminopteridin-6-yl-methyl)]-amino-methyl}-naphthalene
##STR00188##
[0597] To a solution of 6-bromomethyl-2,4-pteridinediamine
hydrobromide (51 mg, 0.2 mmol) in anhydrous N,N dimethylacetamide
was added 1-aminomethyl-naphthalene (31.67 ul, 0.22 mmol). The
reaction mixture was stirred at 50.degree. C. overnight. The crude
product was poured into saturated bicarbonate solution. The
resulted precipitate was collected and purified by preparative
HPLC. 9 mg product was obtained. Yield: 15%; .sup.1H NMR (500 MHz,
DMSO-d6): .delta. 4.0970 (s, 4H), 4.2526 (s, 2H), 7.3530-7.3692
(dd, J.sub.1=7.25 Hz, J.sub.2=7.25 Hz, 2H), 7.439-7.5202 (m, 2H),
7.5414-7.5553 (d, J=6.94 Hz, 1H), 7.67408-7.69065 (d, J=8.285 Hz,
1H), 7.78789-7.7713 (d, J=8.285 Hz, 1H), 8.14819-8.1313 (d, J=8.44
Hz, 1H), 8.7144 (s, 2H), 8.93305 (s, 2H), 9.23424 (s, 21-1);
ESI-MS: m/z 506 (M.sup.++1)
Quinazolines
[0598] General procedure for the 3H-quinazolin-4-one synthesis
[0599] Method 1:
##STR00189##
[0600] Method 2:
##STR00190##
6-bromo-3H-quinazolin-4-one
##STR00191##
[0601] 2-Amino-5-Bromo-benzoic acid (10.817 g, 50 mmol) was
suspended in 70 ml formamide. The mixture was heated at 180.degree.
C. for 7 hrs. The cooled solution was diluted with 100 ml cold
water and filtered. The tan solid was washed with di water and used
for the next step reaction without further purification. 10.2 g
product was obtained. Yield: 90%. .sup.1H NMR (500 MHz, DMSO-d6):
.delta. 7.61430-7.63179 (d, J=8.745 Hz, 1H), 7.94922-7.97149 (dd,
J.sub.1=8.75 Hz, J.sub.2=2.385 Hz, 1H), 8.142421 (s, 1H),
8.19136-8.19609 (d, J=2.365 Hz, 1H); ESI-MS: m/z 225, 227
(M.sup.++1)
6-(2,6-Dimethylphenyl)-3H-quinazolin-4-one
##STR00192##
[0603] To a solution of 6-bromo-3H-quinazolin-4-one (43.1 mg,
0.1915 mmol) dissolved in 2 ml N,N-dimethylacetamide in a 20 ml
vial, 2,6-dimethylphenylboronic acid (114.9 mg, 0.76 mmol)
dissolved in 1 ml ethanol and potassium carbonate (26.7 mg, 0.193
mmol) dissolved in 1 ml water were added. Triphenylphosphine (5 mg,
0.019 mmol) and tris(dibenzylideneacetone)dipalladium(0) (3.5 mg,
3.8 umol) were added to the mixture which refluxed overnight. The
crude product was poured into 50 ml saturated bicarbonate solution
and methylene chloride was used to extract the product. Solvent in
the organic phase was removed under vacuum. The resulted residue
was purified by preparative HPLC. 19.2 mg product was obtained.
Yield: 40%; .sup.1H NMR (500 MHz, DMSO-d6): .delta. 1.96741 (s,
6H), 7.114769-7.16307 (d, J=7.69 Hz, 2H), 7.19260-7.22248 (dd,
J.sub.1=8.62 Hz, J.sub.2=6.31 Hz 1H), 7.60434-7.62503 (dd,
J.sub.1=8.335 Hz, J.sub.2=1.97 Hz, 1H), 7.75179-7.76829 (d, J=8.25
Hz, 1H), 7.81882-7.82258 (d, J=1.88 Hz, 1H), 8.17882 (s, 1H);
ESI-MS: m/z 251 (M.sup.++1)
6-(2,6-Dimethoxlphenyl)-3H-quinazolin-4-one
##STR00193##
[0605] To a solution of 6-bromo-3H-quinazolin-4-one (43.1 mg,
0.1915 mmol) dissolved in 2 ml N,N-dimethylacetamide in a 20 ml
vial, 2,6-dimethylphenylboronic acid (139.4 mg, 0.76 mmol)
dissolved in 1 ml ethanol and potassium carbonate (26.7 mg, 0.193
mmol) dissolved in 1 ml water were added. Triphenylphosphine (5 mg,
0.019 mmol) and tris(dibenzylideneacetone)dipalladium (0) (3.5 mg,
3.8 umol) were added to the mixture which refuxed overnight. The
crude product was poured into 50 ml saturated bicarbonate solution
and methylene chloride was used to extract the product. Solvent in
the organic phase was removed under vacuum. The resulted residue
was purified by preparative HPLC. 38.2 mg product was obtained.
Yield: 71%; .sup.1H NMR (500 MHz, DMSO-d6): .delta. 3.67800 (s,
6H), 6.77555-6.79250 (d, J=8.475 Hz, 1H), 7.33529-7.36895 (dd,
J.sub.1=8.415 Hz, J.sub.2=8.415 Hz 1H), 7.65311 (s, 2H), 7.93672
(s, 1H), 8.13028 (s, 1H); ESI-MS: m/z 283 (M.sup.++1)
6-(2-chloro-6-methoxyphenyl)-3H-quinazolin-4-one
##STR00194##
[0607] To a solution of 6-bromo-3H-quinazolin-4-one (38.9 mg,
0.1728 mmol) dissolved in 2 ml N,N-dimethylacetamide in a 20 ml
vial, 2-chloro-6-methoxy-phenylboronic acid (128.88 mg, 0.6914
mmol) dissolved in 1 ml ethanol and potassium carbonate (26.28 mg,
0.19 mmol) dissolved in 1 ml water were added. Triphenylphosphine
(4.5 mg, 0.017 mmol) and tris(dibenzylideneacetone)dipalladium(0)
(3.2 mg, 3.5 umol) were added to the mixture which refluxed
overnight. The crude product was poured into 5 ml saturated
bicarbonate solution and methylene chloride was used to extract the
product. Solvent in the organic phase was removed under vacuum. The
resulted residue was purified by preparative HPLC. 3.4 mg product
was obtained. Yield: 24.3%; .sup.1H NMR (500 MHz, DMSO-d6): .delta.
3.70812 (s, 3H), 7.13816-7.15637 (dd, J.sub.1=7.945 Hz,
J.sub.2=0.32 Hz, 1H), 7.18430-7.20184 (dd, J.sub.1=7.85 Hz,
J.sub.2=0.92 Hz 1H), 7.40806-7.44074 (dd, J.sub.1=8.205 Hz,
J.sub.2=8.135 Hz, 1H), 7.66531-7.68611 (dd, J.sub.1=8.305 Hz,
J.sub.2=2.04 Hz, 1H), 7.71531-7.73209 (d, J=8.39 Hz, 1H),
7.92946-7.93334 (d, J=1.94 Hz, 1H), 8.16800 (s, 1H); ESI-MS: m/z
287 (M.sup.++1)
6-(2,4,6-trimethylphenyl)-3H-quinazolin-4-one
##STR00195##
[0609] To a solution of 6-bromo-3H-quinazolin-4-one (43.1 mg,
0.1915 mmol) dissolved in 2 ml N,N-dimethylacetamide in a 20 ml
vial, 2,4,6-trimethylphenylboronic acid (114.9 mg, 0.76 mmol)
dissolved in 1 ml ethanol and potassium carbonate (26.7 mg, 0.193
mmol) dissolved in 1 ml water were added. Triphenylphosphine (5 mg,
0.019 mmol) and tris(dibenzylideneacetone)dipalladium (0) (3.5 mg,
3.8 umol) were added to the mixture which refluxed overnight. The
crude product was poured into 50 ml saturated bicarbonate solution
and methylene chloride was used to extract the product. Solvent in
the organic phase was removed under vacuum. The resulted residue
was purified by preparative HPLC. 19.2 mg product was obtained.
Yield: 40%; .sup.1H NMR (500 MHz, DMSO-d6): .delta. 1.96741 (s,
6H), 7.114769-7.16307 (d, J=7.69 Hz, 2H), 7.19260-7.22248 (dd,
J.sub.1=8.62 Hz, J.sub.2=6.31 Hz 1H), 7.60434-7.62503 (dd,
J.sub.1=8.335 Hz, J.sub.2=1.97 Hz, 1H), 7.75179-7.76829 (d, J=8.25
Hz, 1H), 7.81882-7.82258 (d, J=1.88 Hz, 1H), 8.17882 (s, 1H);
ESI-MS: m/z 265 (M.sup.++1)
6-(Naphthalene-1-yl)-3H-quinazolin-4-one
##STR00196##
[0611] To a solution of 6-bromo-3H-quinazolin-4-one (45.2 mg, 0.2
mmol) dissolved in 2 ml N,N-dimethylacetamide in a 20 ml vial,
naphthalene-1-boronic acid (69.4 mg, 0.4 mmol) dissolved in 1 ml
ethanol and potassium carbonate (30.5 mg, 0.22 mmol) dissolved in 1
ml water were added. Tripenylphosphine (5.27 mg, 0.02 mmol) and
tris(dibenzylideneacetone)dipalladium (0) (3.6 mg, 4 umol) was
added to the mixture which refluxed overnight. The crude product
was poured into 50 ml saturated bicarbonate solution and methylene
chloride was used to extract the product. Solvent in the organic
phase was removed under vacuum. The resulted residue was purified
by preparative HPLC. 32.9 mg product was obtained. Yield: 62%;
.sup.1H NMR (500 MHz, DMSO-d6): .delta. 7.52083-7.54615 (m, 2H),
7.56877-7.58461 (dd, J=6.88 Hz, 1H), 7.61224-7.64281 (dd,
J.sub.1=8.255 Hz, J.sub.2=8.285 Hz, 1H), 7.78775-7.804 (d, J=8.125
Hz, 1H), 7.82384-7.84054 (d, J=8.35 Hz, 1H), 7.93472-7.95545 (dd,
J.sub.1=8.365 Hz, J.sub.2=2 Hz, 1H), 8.00847-8.02533 (d, J=8.43 Hz,
1H), 8.03829-8.05347 (d, J=7.59 Hz, 1H), 8.15915-8.16300 (d,
J=1.925 Hz, 1H), 8.19218 (s, 1'-1); ESI-MS: m/z 273 (M.sup.++1)
6-(Naphthalene-2-yl)-3H-quinazolin-4-one
##STR00197##
[0613] To a solution of 6-bromo-3H-quinazolin-4-one (47.1 mg,
0.2093 mmol) dissolved in 2 ml N,N-dimethylacetamide in a 20 ml
vial, naphthalene-1-boronic acid (73 mg, 0.4244 mmol) dissolved in
1 ml ethanol and potassium carbonate (32.7 mg, 0.2366 mmol)
dissolved in 1 ml water were added. Triphenylphosphine (5.5 mg,
0.021 mmol) and tris(dibenzylideneacetone)dipalladium (0) (3.8 mg,
4.1 umol) were added to the mixture which refluxed overnight. The
crude product was poured into 50 ml saturated bicarbonate solution
and methylene chloride was used to extract the product. Solvent in
the organic phase was removed under vacuum. The resulted residue
was purified by preparative HPLC. 26.3 mg product was obtained.
Yield: 46%; .sup.1H NMR (500 MHz, DMSO-d6): .delta. 7.54020-7.58965
(m, 2H), 7.80614-7.82312 (d, J=8.49 Hz, 1H), 7.94743-7.96828 (dd,
J.sub.1=8.505 Hz, J.sub.2=1.91 Hz, 1H), 7.96828-7.98243 (d, J=8.035
Hz, 1H), 8.05455-8.07187 (d, J=8.63 Hz, 1H), 8.16005 (s, 1H),
8.30107-8.3226 (dd, J.sub.1=8.58 Hz, J.sub.2=2.25 Hz, 1H),
8.37163-8.37447 (d, J=1.42 Hz, 1H), 8.50638-8.51090 (d, J=2.26 Hz,
1H); ESI-MS: m/z 273 (M.sup.++1)
6-(4-phenoxy-phenyl)-3H-quinazolin-4-one
##STR00198##
[0615] To a solution of 6-bromo-3H-quinazolin-4-one (44.8 mg, 0.199
mmol) dissolved in 2 ml N,N-dimethylacetamide in a 20 ml vial,
naphthalene-1-boronic acid (85.22 mg, 0.3981 mmol) dissolved in 1
ml ethanol and potassium carbonate (30.26 mg, 0.2198 mmol)
dissolved in 1 ml water were added. Triphenylphosphine (5.2 mg,
0.020 mmol) and tris(dibenzylideneacetone)dipalladium (0) (3.64 mg,
4.0 umol) were added to the mixture which refluxed overnight. The
crude product was poured into 50 ml saturated bicarbonate solution
and methylene chloride was used to extract the product. Solvent in
the organic phase was removed under vacuum. The resulted residue
was purified by preparative HPLC. 25.3 mg product was obtained.
Yield: 41%; .sup.1H NMR (500 MHz, DMSO-d6): .delta. 7.09215-7.12687
(dd, J.sub.1=8.58 Hz, J.sub.2=8.78 Hz, 4H), 7.17733-7.20876 (dd,
J.sub.1=6.48 Hz, J.sub.2=7.375 Hz, 1H), 7.42050-7.45247
(J.sub.1=7.56 Hz, J.sub.2=6.45 Hz, 2H), 7.74247-7.75949 (d, J=8.51
Hz, 1H), 7.79084-7.80838 (dd, J.sub.1=6.73 Hz, J.sub.2=2.08 Hz,
2H), 8.1191-8.1408 (dd, J.sub.1=8.395 Hz, J.sub.2=2.355 Hz, 1H),
8.14531 (s, 1H), 8.31298-8.31761 (d, J=2.315 Hz, 1H); ESI-MS: m/z
315 (M.sup.++1)
6-Bromo-3-(3-hydroxy-propionyl-3H-quinazolin-4-one
##STR00199##
[0617] To a suspension of NaH (60% in mineral oil, 199 mg) in 20 ml
of N,N-dimethylacetamide was added 6-bromo-3H-quinazolin-4-one
(0.9335 mg, 4.148 mmol). The mixture was stirred at room
temperature for 40 mins resulting clear red solution. Acroyl
chloride (471.8 ul, 5.8072 mmol) was added. The solution was heated
at 70.degree. C. for 8 hrs, cooled to room temperature, and poured
into 30 ml of ice water. Methylene chloride added and product was
in the water phase. The water solvent was evaporated under vacuum.
The resulted residue was purified by preparative HPLC. 1.1 g
product was obtained. Yield: 74.7% .sup.1H NMR (500 MHz, DMSO-d6):
.delta. 2.73412-2.76135 (t, J=6.805 Hz, 2H), 4.14197-4.16922 (t,
J=6.815 Hz, 2H), 7.62305-7.64046 (d, J=8.705 Hz, 1H),
7.96596-7.98797 (dd, J.sub.1=8.635 Hz, J.sub.2=2.38 Hz, 1H),
8.2287-8.2335 (d, J=2.4 Hz, 1H), 8.41991 (s, 1H); ESI-MS: m/z 297,
299 (M.sup.++1)
6-(2,6-Dimethylphenyl)-3-(3-hydroxy-propionyl)-3H-quinazolin-4-one
##STR00200##
[0619] To a solution of
6-Bromo-3-(3-hydroxy-propionyl)-3H-quinazolin-4-one (9.8 mg, 0.033
mmol) dissolved in 1 ml N,N-dimethylacetamide in a 20 ml vial,
2,6-dimethylphenyl boronic acid (9.89 mg, 0.066 mmol) dissolved in
0.5 ml ethanol and potassium carbonate (5 mg, 0.036 mmol) dissolved
in 0.5 ml water were added. Triphenylphosphine (0.87 mg, 3.3 umol)
and tris(dibenzylideneacetone)dipalladium(0) (0.6 mg, 0.6 umol)
were added to the mixture which refluxed overnight. The crude
product was poured into 5 ml saturated bicarbonate solution and
methylene chloride was used to extract the product. Solvent in the
organic phase was removed under vacuum. The resulted residue was
purified by preparative HPLC. 5.2 mg product was obtained. Yield:
49%; .sup.1H NMR (500 MHz, DMSO-d6): .delta. 1.96247 (s, 6H),
2.76290-2.79002 (t, J=6.805 Hz, 2H),), 4.15954-4.18664 (t, J=6.785
Hz, 2H), 7.14682-7.7.1621 (d, J=7.64 Hz, 1H), 7.19338-7.21062 (dd,
J.sub.1=8.62 Hz, J.sub.2=6.41 Hz, 1H), 7.60532-7.62604 (dd,
J.sub.1=8.365 Hz, J.sub.2=2.03 Hz, 1H), 7.75204-7.76861 (d, J=8.285
Hz, 1H), 7.84928-7.85312 (d, J=1.92 Hz, 1H), 8.41195 (s, 1H);
ESI-MS: m/z 323 (M.sup.++1)
6-(2-chloro-6-methoxyphenyl)-3-(3-hydroxy-propionyl)-3H-quinazolin-4-one
##STR00201##
[0621] To a solution of
6-Bromo-3-(3-hydroxy-propionyl)-3H-quinazolin-4-one (11.6 mg, 0.039
mmol) dissolved in 1 ml N,N-dimethylacetamide in a 20 ml vial,
2-chloro-6-methoxy-phenylboronic acid (14.55 mg, 0.078 mmol)
dissolved in 0.5 ml ethanol and potassium carbonate (5.92 mg, 0.043
mmol) dissolved in 0.5 ml water were added. Triphenylphosphine (1
mg, 3.8 umol) and tris(dibenzylideneacetone)dipalladium (0) (0.7
mg, 0.78 umol) were added to the mixture which refluxed overnight.
The crude product was poured into 5 ml saturated bicarbonate
solution and methylene chloride was used to extract the product.
Solvent in the organic phase was removed under vacuum. The resulted
residue was purified by preparative HPLC. 3.4 mg product was
obtained. Yield: 24.3%; .sup.1H NMR (500 MHz, DMSO-d6): .delta.
2.75538-2.78226 (t, J=6.835 Hz, 2H), 3.70334 (s, 3H),
4.15877-4.18594 (t, J=6.785 Hz, 2H), 7.13724-7.15535 (dd,
J.sub.1=8.68 Hz, J.sub.2=0.75 Hz, 1H), 7.18337-7.20169 (dd,
J.sub.1=8.375 Hz, J.sub.2=0.885 Hz, 1H), 7.41001-7.44275 (dd,
J.sub.1=8.215 Hz, J.sub.2=8.185 Hz, 1H), 7.66453-7.68523 (dd,
J.sub.1=8.38 Hz, J.sub.2=2.0 Hz, 1H), 7.72 (d, J=8.4 Hz, 1H), 7.96
(d, J=1.9 Hz, 1H), 8.41 (s, 1H); ESI-MS: m/z 359 (M.sup.++1)
2-hydroxy-4-aminoquinazolines
##STR00202##
[0622] 4-Amino-8-bromo-6-nitro-quinazolin-2-ol
##STR00203##
[0623] 2-Amino-3-bromo-5-nitro-benzonitrile (1.9003 g, 7.85 mmol)
was heated with urea (1.8862 g, 31.4 mmol) at 180-185.degree. C.
for 3 hrs. The cooled mixture was powered and treated with
bicarbonate solution, filtered and washed with water. The solid was
the collected and washed with ethanol, ether, and used for the next
step reaction without further purification. 2.0 g product was
obtained. Yield 89%; .sup.1H NMR (500 MHz, DMSO-d6): .delta.
8.44455-8.45011 (d, J=2.78 Hz, 1H), 8.87071-8.87544 (d, J=2.365 Hz,
1H), 9.39866-9.40333 (d, J=2.335 Hz, 1H), 9.50740-9.51282 (d,
J=2.71 Hz, 1H); ESI-MS: 285, 287 (M.sup.++1)
8-Bromo-4-[3-(4-methyl-piperazin-1-yl)-propylamino]-6-nitro-quinazolin-2-o-
l
##STR00204##
[0625] A mixture of 4-amino-8-bromo-6-nitro-quinazolin-2-ol (24.1
mg, 0.0845 mmol), sulfamic acid (16.4 mg, 0.169 mmol) and
1-(3-aminopropyl)-4-methylpiperazine (1 ml) was heated at reflux
for 7 h. The cooled reaction mixture was poured into 10 ml ice
water. The resulting precipitate was collected and purified by
preparative HPLC. 19.2 mg product was obtained. Yield: 40%; .sup.1H
NMR (500 MHz, DMSO-d6): .delta. 1.91521-1.95482 (m, 2H), 2.78103
(s, 8H), 3.16555 (b, 4H), 8.68221-8.68666 (d, J=2.225 Hz, 1H),
9.10824-9.11291 (d, J=2.335 Hz, 1H); ESI-MS: 425, 427
(M.sup.++1)
Preparation of
(6,7-Diphenyl-pteridin-4-yl)-(3-(4-methyl-piperazin-1-yl)-propyl)-amine
##STR00205##
[0627] 6,7-Diphenyl-pteridin-4-ylamine (200 mg, 0.669 mmol) and
sulfamic acid (300 mg, 1.91 mmol) were dissolved in 4 ml
1-(3-aminopropyl)-4-methylpiperazine. The mixture was reflux for
overnight. Preparative HPLC was used to isolated the product. 50 mg
(6,7-Diphenyl-pteridin-4-yl)-(3-(4-methyl-piperazin-1-yl)-propyl)-amine
was obtained. Yield: 17%, ESI-MS: [M+H].sup.+, 441
Representative Synthesis of Compounds of Structure IV
##STR00206##
[0629] A 3-mL reaction flask equipped with a stirring vane and a
teflon cap was charged with the bis(benzil) species (122 mg; 0.324
mmol) and 5,6-diamino-2,4-dihydroxy pyrimidine sulfate (156 mg;
0.649 mmol; 2.00 equiv). The vial was heated to ca. 210.degree. C.
for 2 h and then the contents were poured into 30 mL of ether, the
resulting solid was sonicated vortexed and centrifuged. The
resulting solid was washed 2.times.20 mL of ethyl acetate-ether
(1:1), and dried in a vacuum dessicator resulting in 120 mg (96%)
of an orange solid bis(pteridine). MS (M+H.sup.+: calcd 647; found
647).
Representative Synthesis of Compounds of Structure V
##STR00207##
[0631] A 5-mL, single-necked, round-bottomed flask with a stirring
bar and a septum was charged with 2-aminomethylbenzimidazole (119
mg; 0.500 mmol; 1.00 equiv). It does not dissolve in 3 mL of DMF
even with heating. To this slurry was added isatin (73.8 mg; 0.502
mmol; 1.00 equiv). The solution is a bright orange-yellow. A few
drops of glacial HOAc were added, the reaction was stirred for 15
min, and then sodium cyanoborohydride (62.0 mg; 0.980 mmol; 1.97
equiv). The solution turned a light straw-yellow in 30 min. After
stirring for 2 d at room temperature, the reaction was worked up by
pouring the mixture into 50:50 saturated aqueous sodium
bicarbonate-ice. The white precipitate formed was extracted with
ethylacetate (2.times.20 mL). The combined organic layer was
extracted again with 10 mL satd sodium bicarbonate, dried
(anhydrous Na.sub.2SO.sub.4), filtered, and concentrated by rotary
evaporation to yield an orange-yellow oil that solidified on
standing. The crude was recrystallized from ethylacetate-hexanes to
yield 98.9 mg of an orange foam. MS (M+H.sup.+: calcd 279; found
279)
Example 2
Anti-Cancer Therapy with Vasculostatic Agents
[0632] The following experiments show the use of vasculostatic
agents of the invention alone and in combination with
chemotherapeutic agents for treatment of cancer. FIG. 2 shows the
synergistic results of co-drug therapy utilitizing
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine, sulfate salt,
(compound A--in this example formulated in 50% PEG400:50% water)
illustrated in FIG. 1, with doxorubicin (in this example formulated
in 50% PEG400:50% water). In the experiment shown in FIG. 2,
syngeneic Lewis lung carcinoma cells were injected I.V. in order to
establish lung metastases in Balb/C mice. Beginning 10 days after
cells were injected, doxorubicin (3 mg/kg) and/or
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine, sulfate salt,
(compound A--various doses as shown) was given I.P. every 3 days
for 3 cycles. Animals were sacrificed at day 20, lungs were
collected, and weighed. Net tumor burden is the weight of
tumor-bearing lungs minus the average weight of normal control
lungs. N=5/group, p<0.02. As shown in FIG. 2
6,7-Bis(4-hydroxyphenyl)-pteridin-4-ylamine, sulfate salt (compound
A) had a profound effect on tumor burden in animals, typically
reducing tumor burden by 25% as a stand alone agent or by greater
than 90% in combination with doxorubicin.
[0633] FIG. 3 shows the results of using
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine sulfate salt (compound
A--in this example formulated in 50% PEG400:50% water), and
6,7-diphenyl-pteridine-2,4-diamine (compound B--in this example
formulated in 50% PEG400:50% water) with doxorubicin to treat colon
carcinoma. Syngeneic CT-26 Colon carcinoma cells were injected I.V.
in order to establish lung metastases in Balb/C mice. Beginning 10
days after cells were injected, indicated test agents were given
I.P. every 3 days for 3 cycles. Animals were sacrificed at day 20,
lungs were collected, and weighed. Net tumor burden is the weight
of tumor-bearing lungs minus the average weight of normal control
lungs. N=5/group, p<0.02. In this model, as shown in FIG. 3
6,7-Bis(4-hydroxyphenyl)-pteridin-4-ylamine, sulfate salt (compound
A) typically reduced tumor burden by 35% as a stand alone agent or
by greater than 60% in combination with doxorubicin. Similarly, in
this model, 6,7-diphenyl-pteridine-2,4-diamine (compound B)
typically reduced tumor burden by 35% as a stand alone agent or by
greater than 65% in combination with doxorubicin.
[0634] FIG. 4 illustrates the effects of the compounds of the
present invention for co-drug therapy with docetaxel (Taxotere.RTM.
in this example formulated in 12.5% Cremaphore:12.5% Ethanol:75%
normal saline) as described herein. Syngeneic CT-26 Colon carcinoma
cells were injected I.V. in order to establish lung metastases in
Balb/C mice. Beginning 10 days after cells were injected, indicated
test agents were given I.P. every 3 days for 3 cycles. Animals were
sacrificed at day 20, lungs were collected, and weighed. Net tumor
burden is the weight of tumor-bearing lungs minus the average
weight of normal control lungs. N=5/group, p<0.02.
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine, sulfate salt (compound
A--in this example formulated in 50% PEG400:50% water) and
6,7-diphenyl-pteridine-2,4-diamine (compound B--in this example
formulated in 50% PEG400:50% water) from FIG. 1 are shown in FIG.
4. In this model, as shown in FIG. 4,
6,7-Bis(4-hydroxyphenyl)-pteridin-4-ylamine, sulfate salt (compound
A) typically reduced tumor burden by 25% as a stand alone agent or
by greater than 80% in combination with docetaxel. Similarly, in
this model 6,7-diphenyl-pteridine-2,4-diamine (compound B)
typically reduced tumor burden by 20% as a stand alone agent or by
greater than 70% in combination with doxorubicin.
[0635] FIG. 5 shows a photo of representative lung samples from the
experiment shown in FIG. 3 with 6,7-diphenyl-pteridine-2,4-diamine
(compound B--in this example formulated in 50% PEG400:50% water)
and doxorubicin (in this example formulated in 50% PEG400:50%
water). The tumors in the lungs are apparent in the vehicle
(control) lungs, and the vasculostatic agent plus doxorubicin
treated lungs show a dramatic reduction in tumor burden.
[0636] FIG. 6 illustrates the effect of compounds administered in
conjunction with docetaxel (Taxotere.RTM.--in this example
formulated in 12.5% Cremaphore:12.5% Ethanol:75% normal saline) in
the in vivo model of metastatic colon cancer (CT-26 adenocarcinoma)
described for FIG. 4.
2,3-Bis(3,4-dihydroxyphenyl)-pyrido[2,3-b]pyrazin-6-ylamine
dihydrochloride salt (compound C--in this example formulated in 50%
PEG400:50% water) from FIG. 1 is shown in FIG. 6 as compound C.
N=5/group, p<0.02. In this model, as shown in FIG. 6,
2,3-Bis(3,4-dihydroxyphenyl)-pyrido[2,3-b]pyrazin-6-ylamine
dihydrochloride salt (compound C) typically reduced tumor burden by
65% as a stand alone agent or by greater than 85% in combination
with docetaxel.
[0637] Similarly,
2,3-bis(4-hydroxyphenyl)-pyrido[2,3-b]pyrazin-6-ylamine
dihydrochloride salt inhibited tumor burden alone or with co-drug
therapy using docetaxel (Taxotere.RTM. in this example formulated
in 12.5% Cremaphore:12.5% Ethanol:75% normal saline) as described
herein. Syngeneic CT-26 Colon carcinoma cells were injected I.V. in
order to establish lung metastases in Balb/C mice. Beginning 10
days after cells were injected, indicated test agents were given
I.P. every 3 days for 3 cycles. Animals were sacrificed at day 20,
lungs were collected, and weighed. Net tumor burden is the weight
of tumor-bearing lungs minus the average weight of normal control
lungs. N=5/group, p<0.02.
2,3-Bis(4-hydroxyphenyl)-pyrido[2,3-b]pyrazin-6-ylamine
dihydrochloride salt in 50% PEG400:50% water) typically reduced
tumor burden by 63% as a stand alone agent or by greater than 78%
in combination with docetaxel.
Example 3
Inhibition of Vascular Permeability
[0638] IL-2 is used clinically to treat metastatic melanoma and
renal cell carcinoma and the dose-limiting toxicity for IL-2 is
Vascular Leak Syndrome (VLS). Two representative examples from
distinct chemotype series were selected for initial study in the
reduction of IL-2-induced VLS (see FIG. 1 compounds). The compounds
were pre-screened for in vivo reduction of vascular permeability
and there was no observable gross toxicity as single agents at
20-fold higher doses.
[0639] The results of the studies shown in FIGS. 7-8 indicate that
representative compounds of the invention show inhibition of
vascular leak in vivo. There were no effects on T cell
proliferation in prescribed dose range (see FIGS. 10-11) and no
effects on anti-tumor activity of IL-2 (melanoma model; see FIG.
9). The following experiments exemplify the results for co-drug
therapy.
[0640] BalbC mice were given 9 injections of the indicated dose of
murine IL-2 (in this example formulated in saline with 5% bovine
serum albumin) and/or invention compounds over a period of 4 days.
Animals were then sacrificed followed by collection, blotting and
weighing (wet weight) of heart, lungs, and spleen. Organs were then
dried at 80.degree. C. for 24 hours and weighed (dry weight).
N=5/group, p<0.02. N-(2-(1H-indol-2-yl)-phenyl)-phthalamic acid
(compound D--in the 1 mg/kg range, in this example formulated in
50% PEG400:50% water) and
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine, sulfate salt
(compound E--in the 0.1 mg/kg range, in this example formulated in
50% PEG400:50% water) typically reduced VLS in the heart by
>100%. The results are shown in FIG. 7.
[0641] BalbC mice were given 9 injections of the indicated dose of
murine IL-2 and/or invention compounds over a period of 4 days.
Animals were then sacrificed followed by collection, blotting and
weighing (wet weight) of heart, lungs, and spleen. Organs were then
dried at 80.degree. C. for 24 hours and weighed (dry weight).
N=5/group, p<0.02. N-(2-(1H-indol-2-yl)-phenyl)-phthalamic acid
(compound D--in the 1 mg/kg range, in this example formulated in
50% PEG400:50% water) and
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine, sulfate salt
(compound E--in the 0.1 mg/kg range, in this example formulated in
50% PEG400:50% water) typically reduced VLS in the spleen by
>100%. The results are shown in FIG. 8.
[0642] Syngeneic B16 melanoma cells were injected I.V. in order to
establish lung metastases in C57 mice. Beginning 10 days after
cells were injected, 100,000 U of IL-2 and/or indicated invention
compounds were given I.P. every 8 hours for 5 days. Animals were
sacrificed at day 18, lungs were collected and scored using image
analysis software. N=5/group, p<0.02.
N-(2-(1H-indol-2-yl)-phenyl)-phthalamic acid (compound D--in the 1
mg/kg range, in this example formulated in 50% PEG400:50% water)
and 6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine, sulfate salt
(compound E--in the 0.1 mg/kg range, in this example formulated in
50% PEG400:50% water) typically had no significant impact on the
anti-tumor activity of IL-2. Invention compound concentrations are
listed in parenthesis in mg/kg while IL-2 concentration is given in
parenthesis kilounits. The results are shown in FIG. 9.
[0643] An IL-2 dependent human T cell line, CTLL2, was used to
evaluate IL-2 dependent proliferation over 96 hours in the presence
of 50 pg of human recombinant IL-2 (R&D Systems) and the
indicated compounds using the XTT assay.
N-(2-(1H-indol-2-yl)-phenyl)-phthalamic acid (compound D--in the 1
mg/kg range, in this example formulated in 50% PEG400:50% water)
typically had no significant impact on IL-2 induced T-cell
proliferation. The results are shown in FIG. 10.
[0644] An IL-2 dependent human T cell line, CTLL2, was used to
evaluate IL-2 dependent proliferation over 96 hours in the presence
of 50 pg of human recombinant IL-2 (R&D Systems) and the
indicated compounds using the XTT assay.
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine, sulfate salt
(compound E--in the 0.1 mg/kg range, in this example formulated in
50% PEG400:50% water) typically had no significant impact on IL-2
induced T-cell proliferation in the therapeutic range (<1
.mu.M). The results are shown in FIG. 11.
[0645] Thus, representative examples from two distinct chemotype
series in the present application (shown in FIG. 1) indicate that,
for example, N-(2-(1H-indol-2-yl)-phenyl)-phthalamic acid (compound
D--in the 1 mg/kg range, in this example formulated in 50%
PEG400:50% water) and
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine, sulfate salt
(compound E--in the 0.1 mg/kg range, in this example formulated in
50% PEG400:50% water), are effective in reducing VLS by 80-100% in
vivo.
[0646] Both of the exemplary compounds performed well in important
initial tests, including 1) inhibition of VLS at normal and
elevated doses of IL-2; 2) no interference with IL-2 mediated
anti-tumor activity; 3) no inhibition of IL-2 induced T cell
proliferation in the likely therapeutic dose range; and 4) neither
compound elicited gross observable toxicity. These results indicate
that invention compounds could be used in conjunction with IL-2 to
prevent dose-limiting VLS and thereby increase the clinical
application and therapeutic dose range of IL-2.
[0647] Acute Respiratory Distress Syndrome (ARDS) is an acute,
severe injury to most or all of both lungs causing fluid leak into
the lungs. Patients with ARDS experience severe shortness of breath
and often require mechanical ventilation (life support) because of
respiratory failure. ARDS has also been called some of the
following terms: Non-cardiogenic pulmonary edema;
Increased-permeability pulmonary edema; Stiff lung; Shock lung;
Adult respiratory distress syndrome; Acute respiratory distress
syndrome. Two representative compounds of the invention were
selected for initial study in the reduction of ARDS.
[0648] NIH Swiss mice were given an intraperitoneal injection of
1.5 mg/kg Oleic Acid of (in this example formulated in saline)
and/or invention compounds. Four hours subsequent to injection
animals were sacrificed followed by collection, blotting and
weighing (wet weight) of the lungs. Lungs were then dried at
80.degree. C. for 24 hours and weighed (dry weight). N=4/group,
6,7-bis(3-hydroxyphenyl)-pteridine-2,4-diamine, sulfate salt
(compound E--in the 0.5 mg/kg range, in this example formulated in
50% PEG400:50% water) typically reduced ARDS-induced edema by
>50% while
4-[4-amino-6-(3,4-dihydroxyphenyl)pteridin-7-yl]benzene-1,2-diol
(compound F--in the 0.5 mg/kg range, in this example formulated in
50% PEG400:50% water) typically reduced ARDS-induced edema by
>100%. The results are shown in FIG. 12.
Example 4
Inhibition of VEGF-Induced Edema
[0649] Miles Assay Data
[0650] A rodent model of vascular edema, the Miles assay, was used
to screen compounds for their ability to inhibit VEGF-induced
edema. The table below presents several examples drawn from these
studies, in which compounds cited in this application successfully
inhibited edema formation.
TABLE-US-00027 Dose Score Treatment (mg/kg BW) (scale of 0-12)
Vehicle 12 4-{[(2,4-Diamino-pteridin-6-ylmethyl)-amino]- 5 mg/kg 4
methyl}-benzene-1,2-diol 4-(2,4-Diamino-pteridin-6-yl)-phenol
(sulfate salt) 5 mg/kg 2
2-[2-(1H-Indol-2-yl)-phenyl]-isoindole-1,3-dione 1.5 mg/kg 3
##STR00208## 1.5 mg/kg 3
6,7-Bis-(3-hydroxy-phenyl)-pteridine-2,4-diol 1.5 mg/kg 3
3-(4-Hydroxy-phenyl)-N-[2-(1H-indol-2-yl)- 1.5 mg/kg 2
phenyl]-propionamide 2-(4-Hydroxy-phenyl)-N-[2-(1H-indol- 1.5 mg/kg
2 2-yl)-phenyl]-acetamide 2-(3,4-Dihydroxy-phenyl)-N-[2-(1H- 0.5
mg/kg 7 indol-2-yl)-phenyl]-acetamide
N-[2-(2,3-Dihydro-1H-indol-2-yl)- 0.5 mg/kg 5
phenyl]-2-hydroxy-benzamide 3-[2-(1H-Indol-2-yl)-phenylcarbamoyl]-
0.5 mg/kg 5 pyridine-2-carboxylic acid
2-Hydroxy-5-(6-phenyl-pteridin-4-ylamino)- 0.5 mg/kg 6
benzenesulfonic acid 5-(6-Phenyl-pteridin-4-ylamino)-quinolin-8-ol
0.5 mg/kg 5 hydrochloride salt
3,4-Dihydroxy-N-[2-(1H-indol-2-yl)-phenyl]- 0.1 mg/kg 6 benzamide
6-{[(Pyridin-2-ylmethyl)-amino]-methyl}- 0.1 mg/kg 4
pteridine-2,4-diamine 6-{[(Naphthalen-2-ylmethyl)-amino]-methyl}-
0.1 mg/kg 4 pteridine-2,4-diamine
2,3-(3,4-Dihydroxyphenyl)-pyrido[3,4- 0.01 mg/kg 6
b]pyrazin-8-ylamine 3-[2,4-diamino-6-(3-hydroxyphenyl)pteridin-7- 1
mg/kg 4 yl]phenol dihydrochloride salt
3-[2,4-diamino-6-(3-hydroxyphenyl)pteridin-7- 0.1 mg/kg 4 yl]phenol
dihydrochloride salt 3-[2,4-diamino-6-(3-hydroxyphenyl)pteridin-7-
0.01 mg/kg 3 yl]phenol dihydrochloride salt
4-[4-amino-6-(3,4-dihydroxyphenyl)pteridin-7- 1 mg/kg 5
yl]benzene-1,2-diol chloride salt
4-[4-amino-6-(3,4-dihydroxyphenyl)pteridin-7- 0.1 mg/kg 3
yl]benzene-1,2-diol chloride salt
4-[4-amino-6-(3,4-dihydroxyphenyl)pteridin-7- 0.01 mg/kg 6
yl]benzene-1,2-diol chloride salt
Sprague-Dawley rats were first injected IV with vehicle alone or
test agent, followed by IV injection of Evans blue dye, followed by
intradermal injections of saline and VEGF (200 ng/injection site)
along both shaved flanks. After 45 min, intradermal injection sites
were photographed and then scored by a blinded observer for
extravasation of Evans blue dye into the dermis (dermal bluing)
according to a 4 point scoring system (3=maximal bluing,
.gtoreq.75% of response in vehicle-treated animals; 2=medium
bluing, >25% but <75% of vehicle-treated animals; 1=minimal
bluing, .ltoreq.25% of vehicle-treated animals; 0=bluing equivalent
to saline injection sites on same animal). Individual scores for 4
injection sites (from 2 separate animals) were summed and are shown
as a scale of 0-12, with a lower score indicating the greater
anti-edema activity; note that all vehicle-treated groups score a
value of 12, based on the scoring system outlined above.
[0651] The ability of test agents to influence edema induced by
agonists other than VEGF was also tested. Compounds cited in this
application inhibited edema formation induced using histamine as an
agonist, for example, as shown below.
TABLE-US-00028 Score Score with VEGF with histamine Dose as agonist
as agonist Treatment (mg/kg BW) (scale of 0-12) (scale of 0-12)
Vehicle 12 12 6,7-bis(4- 1.5 mg/kg 4 3 hydroxyphenyl)-
pteridin-4-ylamine sulfate salt 6,7-Diphenyl- 1.5 mg/kg 3 4
pteridin-4-ol 3,4,5-Trihydroxy- 1.5 mg/kg 4 7 N-[2-(1H-indol-2-
yl)-phenyl]- benzamide 3,4,5-Trihydroxy- 1.5 mg/kg 5 7
N-(1H-indol-2-yl)- benzamide
The ability of test agent to influence vascular edema was tested as
above, except that the ability to block edema was tested using
either VEGF or histamine as the agonist (200 ng and 10
.mu.g/injection site, respectively).
Example 5
Reduction of Myocardial Infarction
[0652] Myocardial Infarct Data
[0653] A rodent model of acute myocardial infarct, in which the
proximal left anterior descending coronary artery (LAD) is occluded
for 60 min followed by reperfusion, was used to determine whether
test agents reduced infarct size at 24 hours. Several examples of
the compounds cited in this application significantly reduced
infarct size as compared to controls.
TABLE-US-00029 Dose Infarct (% AAR, % Infarct Study # Treatment
(mg/kg BW) mean .+-. SEM) reduction 1 Vehicle 75.9 .+-. 1.8
6,7-bis(4- 1.5 60.6 .+-. 1.8 20% hydroxyphenyl)-pteridin- 4-ylamine
sulfate salt 2 Vehicle 54.0 .+-. 2.9 6,7-bis(3,4- 1.5 36.3 .+-. 6.3
33% dihydroxyphenyl)- pteridine-2,4,-diamine, hydrochloride salt 3
Vehicle 54.0 .+-. 2.9 3-[2,4-diamino-6-(3- 1.0 46.4 .+-. 2.6 Not
significant hydroxyphenyl)pteridin- 7-yl] phenol dihydrochloride
salt 3-[2,4-diamino-6-(3- 0.1 37.7 .+-. 5.8 30%
hydroxyphenyl)pteridin- 7-yl] phenol dihydrochloride salt 4 Vehicle
61.9 .+-. 3.1 4-[4-amino-6-(3,4- 1.0 mg/kg 40.1 .+-. 2.0 35%
dihydroxyphenyl)pteridin- 7-yl]benzene-1,2-diol chloride salt
4-[4-amino-6-(3,4- 0.1 mg/kg 37.1 .+-. 2.6 40%
dihydroxyphenyl)pteridin- 7-yl]benzene-1,2-diol chloride salt
6,7-Bis(3- 1.0 mg/kg 39.1 .+-. 7.5 37% hydroxyphenyl)-pteridine-
4-ylamine hydrochloride salt 6,7-Bis(3- 0.1 mg/kg 39.1 .+-. 4.2 37%
hydroxyphenyl)-pteridine- 4-ylamine hydrochloride salt 5 Vehicle
54.9 .+-. 3.1 3-[2,4-Diamino-6-(3- 0.5 mg/kg 31.6 .+-. 6.2 42%
hydroxyphenyl)pteridin- 7-yl]phenol dibromide salt 6,7-bis(3- 0.5
mg/kg 37.8 .+-. 4.5 31% hydroxyphenyl)-pteridine- 2,4-diamine (PF1)
6,7-bis(3- 0.5 mg/kg 35.4 .+-. 1.8 35% hydroxyphenyl)-pteridine-
2,4-diamine (PF2) 6,7-bis(3- 0.5 mg/kg 38.7 .+-. 5.3 29%
hydroxyphenyl)-pteridine- 2,4-diamine (PF5)
Myocardial infarcts were created in Sprague-Dawley rats (200-300 g
body weight) by a 60 min occlusion of the LAD followed by LAD
reperfusion. At 90 mM post-reperfusion, either vehicle alone or
test agents were injected IV. At 24 hr post-treatment, the ischemic
zone (area at-risk, AAR) was delineated by re-ligation of the LAD
followed by IV injection of alkali blue dye, after which hearts
were sectioned along the short axis and stained using
triphenyltetrazolium chloride to delineate viable from infarcted
myocardium. Photographic images were then analyzed using
morphometric software to calculate infarct area as a percent of the
at-risk area. Study 1: Group sizes N=5-6;
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine sulfate salt differs
from vehicle control (P<0.0005). Study 2: Group sizes N=5;
6,7-bis(3,4-dihydroxyphenyl)-pteridine-2,4-diamine hydrochloride
salt differs from vehicle control (P<0.035). Study 3: Group
sizes N=3-5; 3-[2,4-diamino-6-(3-hydroxyphenyl)pteridin-7-yl]phenol
dihydrochloride salt at 0.1 mg/kg differs from vehicle control
(P<0.03). Study 4: Group sizes N=4-5; all
4-[4-amino-6-(3,4-dihydroxyphenyl)pteridin-7-yl]benzene-1,2-diol
chloride salt and 6,7-Bis(3-hydroxyphenyl)-pteridine-4-ylamine
hydrochloride salt treatment groups differ from vehicle control
(P<0.02). Study 5:
3-[2,4-Diamino-6-(3-hydroxyphenyl)pteridin-7-yl]phenol dibromide
salt was delivered in 8% PEG400 (Vehicle), while
6,7-Bis(3-hydroxyphenyl)-pteridine-2,4-diamine was delivered as one
of three product formulations (PF1=2.8%
hydroxypropyl-.alpha.-cyclodextrin, 1.84% PEG400, and 0.009% EDTA
in 20 mM pH 3 citrate buffer; PF2=1.8% hydroxypropyl-S-cyclodextrin
and 0.06% polyvinylpyrrolidone in 20 mM pH 3 citrate buffer;
PF3=0.8% sulfonbutyl ether-.alpha.-cyclodextrin and 0.03%
polyvinylpyrrolidone in 20 mM pH 3 citrate buffer). Group sizes
N=5-6; all treatment groups differ from vehicle control
(P<0.05).
[0654] The following studies were performed as described above,
except that the timing of
3-[2,4-diamino-6-(3-hydroxyphenyl)pteridin-7-yl]phenol
dihydrochloride salt administration (at 0.1 mg/kg) was varied. In
one group, 3-[2,4-diamino-6-(3-hydroxyphenyl)pteridin-7-yl]phenol
dihydrochloride salt was administered at both 60 and 240 min
post-occlusion.
TABLE-US-00030 Administration time (min post- Infarct (% AAR, %
Infarct Treatment occlusion) mean .+-. SEM) reduction Vehicle 60
54.0 .+-. 2.9 3-[2,4-diamino-6-(3- 60 21.6 .+-. 5.7 60%
hydroxyphenyl)pteridin- 7-yl] phenol dihydrochloride salt
3-[2,4-diamino-6-(3- 120 18.8 .+-. 5.6 65% hydroxyphenyl)pteridin-
7-yl] phenol dihydrochloride salt 3-[2,4-diamino-6-(3- 240 19.1
.+-. 4.0 65% hydroxyphenyl)pteridin- 7-yl] phenol dihydrochloride
salt 3-[2,4-diamino-6-(3- 60 and 240 24.2 .+-. 4.9 55%
hydroxyphenyl)pteridin- 7-yl] phenol dihydrochloride salt
Group sizes N=4-5; all
3-[2,4-diamino-6-(3-hydroxyphenyl)pteridin-7-yl]phenol
dihydrochloride salt treatment groups differ from vehicle control
(P<0.001).
[0655] Stroke Data
[0656] A rodent model of cerebral stroke, in which the middle
cerebral artery is permanently occluded, was used to determine
whether test agents reduced infarct size at 24 hours. Several
examples of the compounds cited in this application significantly
reduced infarct size as compared to controls, and to a greater
degree than two commercially available compounds (PP1 and SU6656)
described in the literature as Src kinase inhibitors.
TABLE-US-00031 Infarct area in mm.sup.3 % Infarct Study # Treatment
(mean .+-. SEM) reduction 1 Vehicle 42.4 .+-. 6.25 -- PP1 35.4 .+-.
6.4 Not significant SU6656 24.3 .+-. 5.3 Not significant
6,7-Di-pyridin-2-yl- 27.2 .+-. 2.63 Not significant
pteridin-4-ylamine 6,7-Diphenyl-pteridine-2,4- 20.2 .+-. 4.19 52%
diol N-(2-(1H-Indol-2-yl)- 15.6 .+-. 5.16 63% phenyl)-phthalamic
acid 2 Vehicle 39.0 .+-. 5.0 -- 6,7-bis(4- 18.3 .+-. 2.6 53%
hydroxyphenyl)-pteridin- 4-ylamine, sulfate salt
[0657] Cerebral strokes were created in mice by permanent ligation
of the middle cerebral artery using a cauterizing tool, followed 60
min later by IV injection of either vehicle alone (50% PEG400 in
water) or test agents (at 1 mg/kg BW). Twenty four hours later,
brains were sectioned and stained using triphenyltetrazolium
chloride to delineate viable from infarcted tissue. Photographic
images were then analyzed using morphometric software to calculate
infarct area. [0658] Study 1: Group sizes N=5-6; the
6,7-diphenyl-pteridine-2,4-diol and
N-(2-(1H-indol-2-yl)-phenyl)-phthalamic acid groups differ from
vehicle control (P<0.05 and P<0.01, respectively). [0659]
Study 2: Group sizes N=6-7; the
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine, sulfate salt group
differs from vehicle control (P<0.006).
Example 6
Inhibition of Src-Family Kinases, c-Src and Yes
[0660] The ability of compounds to inhibit the activity of two
Src-family kinases (c-Src and Yes) was directly tested. The table
below presents data for several compounds, which in most cases
inhibited one or both kinases at concentrations of .ltoreq.10
.mu.M.
TABLE-US-00032 Src kinase Yes kinase Compound (IC.sub.50 value)
(IC.sub.50 value) 6,7-bis(3- 27.6 .mu.M 3.8 .mu.M
hydroxyphenyl)-pteridine- 2-amine 6,7-bis(3,4- 2.6 .mu.M 1.1 .mu.M
dihydroxyphenyl)- pteridine-2,4-diamine, hydrochloride salt
2,3-(3,4- 1.6 .mu.M 1.0 .mu.M Dihydroxyphenyl)-
pyrido[3,4-b]pyrazin-8- ylamine 4-[4-amino-6-(3,4- 1.3 .mu.M ND
dihydroxyphenyl)pteridin- 7-yl]benzene-1,2-diol chloride salt
6,7-Bis-(3,4-dihydroxy- 1.8 .mu.M 0.9 .mu.M
phenyl)-pteridine-2,4-diol 3,4-Dihydroxy-N-[2-(1H- 337 nM 303 nM
indol-2-yl)-phenyl]- benzamide 2,3-Bis(3,4- 1.3 .mu.M 756 nM
dihydroxyphenyl)- pyrido[2,3-b]pyrazin-6- ylamine dihydrochloride
salt 6,7-Bis(3- 10.0 .mu.M 6.3 .mu.M hydroxyphenyl)-pteridine-
4-ylamine hydrochloride salt 4-[4-amino-6-(3,4- 0.8 .mu.M ND
dihydroxyphenyl)pteridin- 7-yl]benzene-1,2-diol methanesulfonate
3-(3-Amino- 12.0 .mu.M 6.8 .mu.M benzo[1,2,4]triazin-7-yl)- phenol
7-Naphthalen-1-yl- 0.9 .mu.M 9.3 .mu.M benzo[1,2,4]triazin-3-
ylamine 6,7-Bis(3- 8.8 .mu.M ND hydroxyphenyl)-pteridine- 4-ylamine
hydrobromide salt 7-(2-Trifluoromethyl- 9.2 .mu.M 7.0 .mu.M
phenyl)- benzo[1,2,4]triazin-3- ylamine [7-(2,6-Dimethyl-phenyl)-
925 nM 822 nM benzo[1,2,4]triazin-3-yl]- phenyl-amine
[7-(2,6-Dimethyl-phenyl)- 294 nM ND 5-methyl-
benzo[1,2,4]triazin-3-yl]- phenyl-amine 4-[(Phenyl-pteridin-4- 420
nM ND ylamino)-methyl]- benzene-1,2-diol 4-[2-(6-Phenyl-pteridin-4-
317 nM ND ylamino)-ethyl]benzene- 1,2-diol
Kinase reactions were conducted in 96-well plates by combining
recombinant human c-Src or Yes (280 ng/well, Panvera, Madison
Wis.), ATP (3 .mu.M), a tyrosine kinase substrate (PTK2, 250 .mu.M,
Promega Corp., Madison Wis.), and test agents (at concentrations
ranging from 1 nM to 100 .mu.M); the buffer used was Src kinase
reaction buffer (Upstate USA, Lake Placid N.Y.). After reacting at
90 minutes at room temperature, residual ATP was determined using a
luciferase-based assay (KinaseGlo, Promega Corp.) as a measure of
kinase activity. Data from four wells were then averaged and used
to determine IC.sub.50 values for the test compounds (Prism
software package, GraphPad Software, San Diego Calif.). ND: not
determined.
Example 7
Effects of Invention Compounds on Angiogenesis
[0661] Referring to FIGS. 13 and 14, a murine model of angiogenesis
was used to screen compounds for their capacity to inhibit
angiogenesis. The graph presents representative examples of
compounds cited in this application which successfully inhibited
angiogenesis in vivo. In the graph, compound A is
6,7-bis(4-hydroxyphenyl)-pteridin-4-ylamine sulfate salt. Athymic
WeHi (nu/nu) mice were first injected with 400 .mu.ls of an
ice-cold tumor-derived extracellular matrix substrate, matrigel
(Becton-Dickinson) infused with 400 ng/ml of bFGF or VEGF (R&D
Systems) which rapidly solidifies into a subdermal plug at body
temperature. Mice were subsequently injected intaperitoneally with
10 mg/kg of the indicated compounds bid for four days. On the
fourth day mice were injected intravenously with 0.5 mgs of a
FITC-conjugated endothelial specific lectin (Banderiea Simplifica,
Vector Laboratories). Twenty minutes after injection of the lectin,
mice were euthanized, matrigel plugs were then extracted,
solublized in PBS with mechanical grinding and the fluorescent
content of individual plugs was quantified. Values shown are
normalized to control values from groups of 5.
[0662] Although the invention has been described with reference to
the presently preferred embodiment, it should be understood that
various modifications can be made without departing from the spirit
of the invention. Accordingly, the invention is limited only by the
following claims.
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