U.S. patent application number 11/136242 was filed with the patent office on 2005-10-06 for quinazoline derivatives as medicaments.
This patent application is currently assigned to Scios, Inc.. Invention is credited to Chakravarty, Sarvajit, Dugar, Sundeep, Lewicki, John A., Liu, David Y., Perumattam, John J., Schreiner, George F..
Application Number | 20050220784 11/136242 |
Document ID | / |
Family ID | 22497802 |
Filed Date | 2005-10-06 |
United States Patent
Application |
20050220784 |
Kind Code |
A1 |
Chakravarty, Sarvajit ; et
al. |
October 6, 2005 |
Quinazoline derivatives as medicaments
Abstract
The invention is directed to methods to inhibit TGF-.beta.
and/or p38-.alpha. kinase using compounds of the formula 1 or the
pharmaceutically acceptable salts thereof wherein R.sup.3 is a
noninterfering substituent; each Z is CR.sup.2 or N, wherein no
more than two Z positions in ring A are N, and wherein two adjacent
Z positions in ring A cannot be N; each R.sup.2 is independently a
noninterfering substituent; L is a linker; n is 0 or 1; and Ar' is
the residue of a cyclic aliphatic, cyclic heteroaliphatic, aromatic
or heteroaromatic moiety optionally substituted with 1-3
noninterfering substituents.
Inventors: |
Chakravarty, Sarvajit;
(Mountain View, CA) ; Dugar, Sundeep; (San Jose,
CA) ; Perumattam, John J.; (Los Altos, CA) ;
Schreiner, George F.; (Los Altos, CA) ; Liu, David
Y.; (Palo Alto, CA) ; Lewicki, John A.; (Los
Gatos, CA) |
Correspondence
Address: |
MORRISON & FOERSTER LLP
3811 VALLEY CENTRE DRIVE
SUITE 500
SAN DIEGO
CA
92130-2332
US
|
Assignee: |
Scios, Inc.
Fremont
CA
|
Family ID: |
22497802 |
Appl. No.: |
11/136242 |
Filed: |
May 23, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11136242 |
May 23, 2005 |
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09972582 |
Oct 5, 2001 |
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6903096 |
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09972582 |
Oct 5, 2001 |
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09383825 |
Aug 27, 1999 |
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6476031 |
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09383825 |
Aug 27, 1999 |
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09141916 |
Aug 28, 1998 |
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6184226 |
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Current U.S.
Class: |
424/130.1 ;
514/171; 514/262.1; 514/264.1 |
Current CPC
Class: |
A61P 17/02 20180101;
A61P 13/12 20180101; A61P 7/00 20180101; C07D 401/12 20130101; A61P
9/04 20180101; A61P 19/02 20180101; A61P 9/00 20180101; C07D 239/94
20130101; A61P 1/18 20180101; C07D 471/04 20130101; A61K 31/519
20130101; A61P 43/00 20180101; A61P 9/10 20180101; A61P 11/00
20180101; A61P 29/00 20180101; A61P 1/16 20180101 |
Class at
Publication: |
424/130.1 ;
514/262.1; 514/264.1; 514/171 |
International
Class: |
A61K 039/395; A61K
031/519 |
Claims
1. A pharmaceutical composition for treating conditions
characterized by enhanced p38-.alpha. activity and/or enhanced
TGF-.beta. activity which composition comprises a therapeutically
effective amount of a compound of the formula 22or the
pharmaceutically acceptable salts thereof wherein R.sup.3 is a
noninterfering substituent; each Z is CR.sup.2 or N, wherein no
more than two Z positions in ring A are N, and wherein two adjacent
Z positions in ring A cannot be N; each R.sup.2 is independently a
noninterfering substituent; L is a linker; n is 0 or 1; and Ar' is
the residue of a cyclic aliphatic, cyclic heteroaliphatic, aromatic
or heteroaromatic moiety optionally substituted with 1-3
noninterfering substituents in admixture with at least one
pharmaceutically acceptable excipient.
2. The composition of claim 1 which further contains an additional
therapeutic agent.
3. The composition of claim 2 wherein said additional therapeutic
agent is a corticosteroid, a monoclonal antibody, or an inhibitor
of cell division.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of U.S. patent application
Ser. No. 09/972,582, filed on Oct. 5, 2001, which has been allowed,
and which is a continuation of U.S. Ser. No. 09/383,825 filed Aug.
27, 1999, which is a continuation in part of U.S. Ser. No.
09/141,916 filed Aug. 28, 1998. The contents of these applications
are incorporated herein by reference.
FIELD OF THE INVENTION
[0002] The invention relates to treating various disorders
associated with enhanced activity of kinase p38-.alpha. and/or
transforming growth factor beta (TGF-.beta.). More specifically, it
concerns compounds that are related to quinazoline as useful in
these methods.
BACKGROUND ART
[0003] A large number of chronic and acute conditions have been
recognized to be associated with perturbation of the inflammatory
response. A large number of cytokines participate in this response,
including IL-1, IL-6, IL-8 and TNF. It appears that the activity of
these cytokines in the regulation of inflammation rely at least in
part on the activation of an enzyme on the cell signaling pathway,
a member of the MAP kinase family generally known as p38 and
alternatively known as CSBP and RK. This kinase is activated by
dual phosphorylation after stimulation by physiochemical stress,
treatment with lipopolysaccharides or with proinflammatory
cytokines such as IL-1 and TNF. Therefore, inhibitors of the kinase
activity of p38 are useful antiinflammatory agents.
[0004] Transforming growth factor-beta (TGF-.beta.) denotes a
family of proteins, TGF-.beta.1, TGF-.beta.2, and TGF-.beta.3,
which are pleiotropic modulators of cell growth and
differentiation, embryonic and bone development, extracellular
matrix formation, hematopoiesis, immune and inflammatory responses
(Roberts and Sporn Handbook of Experimental Pharmacology (1990)
95:419-58; Massague et al. Ann Rev Cell Biol (1990) 6:597-646).
Other members of this superfamily include activin, inhibin, bone
morphogenic protein, and Mullerian inhibiting substance. TGF-.beta.
initiates an intracellular signaling pathway leading ultimately to
the expression of genes that regulate the cell cycle, control
proliferative responses, or relate to extracellular matrix proteins
that mediate outside-in cell signaling, cell adhesion, migration
and intercellular communication.
[0005] Therefore, inhibitors of the TGF-.beta. intracellular
signaling pathway are useful treatments for fibroproliferative
diseases. Specifically, fibroproliferative diseases include kidney
disorders associated with unregulated TGF-.beta. activity and
excessive fibrosis including glomerulonephritis (GN), such as
mesangial proliferative GN, immune GN, and crescentic GN. Other
renal conditions include diabetic nephropathy, renal interstitial
fibrosis, renal fibrosis in transplant patients receiving
cyclosporin, and HIV-associated nephropathy. Collagen vascular
disorders include progressive systemic sclerosis, polymyositis,
scleroderma, dermatomyositis, eosinophilic fascitis, morphea, or
those associated with the occurrence of Raynaud's syndrome. Lung
fibroses resulting from excessive TGF-.beta. activity include adult
respiratory distress syndrome, idiopathic pulmonary fibrosis, and
interstitial pulmonary fibrosis often associated with autoimmune
disorders, such as systemic lupus erythematosus and scleroderma,
chemical contact, or allergies. Another autoimmune disorder
associated with fibroproliferative characteristics is rheumatoid
arthritis.
[0006] Eye diseases associated with a fibroproliferative condition
include retinal reattachment surgery accompanying proliferative
vitreoretinopathy, cataract extraction with intraocular lens
implantation, and post glaucoma drainage surgery.
[0007] PCT applications WO98/06715, WO98/07425, and WO 96/40143,
all of which are incorporated herein by reference, describe the
relationship of p38 kinase inhibitors with various disease states.
As mentioned in these applications, inhibitors of p38 kinase are
useful in treating a variety of diseases associated with chronic
inflammation. These applications list rheumatoid arthritis,
rheumatoid spondylitis, osteoarthritis, gouty arthritis and other
arthritic conditions, sepsis, septic shock, endotoxic shock,
Gram-negative sepsis, toxic shock syndrome, asthma, adult
respiratory distress syndrome, stroke, reperfusion injury, CNS
injuries such as neural trauma and ischemia, psoriasis, restenosis,
cerebral malaria, chronic pulmonary inflammatory disease,
silicosis, pulmonary sarcosis, bone resorption diseases such as
osteoporosis, graft-versus-host reaction, Crohn's Disease,
ulcerative colitis including inflammatory bowel disease (IBD) and
pyresis.
[0008] The above-referenced PCT applications disclose compounds
which are p38 kinase inhibitors said to be useful in treating these
disease states. These compounds are either imidazoles or are
indoles substituted at the 3- or 4-position with a piperazine ring
linked through a carboxamide linkage. Additional compounds which
are conjugates of piperazines with indoles are described as
insecticides in WO97/26252, also incorporated herein by
reference.
[0009] The compounds of the invention are quinazoline derivatives.
Other quinazoline compounds for other uses have been described.
U.S. Pat. No. 5,721,237 assigned to Rhone-Poulenc Rorer is directed
to methods for selective treatment of cell growth and
differentiation characterized by activity of human epidermal growth
factor receptor type II using quinazoline substituted only in the
4-position with an aromatic moiety optionally coupled to the
quinazoline through a linking moiety. U.S. Pat. No. 4,480,883
describes compounds that exhibit tyrosine kinase inhibition
activity wherein the heterocyclic portion of a quinazoline or other
fused ring nitrogen-containing aromatic system is substituted only
once with an aromatic moiety, again optionally coupled through a
linker. U.S. Pat. No. 5,616,582 assigned to Zeneca describes
tyrosine kinase inhibitors which are quinazolines linked through an
amino group at the 4-position to a substituted or unsubstituted
phenyl. These compounds contain no substituents at position 2. U.S.
Pat. No. 5,475,001 also assigned to Zeneca describes similar
compounds with the same activity. U.S. Pat. No. 5,430,148 assigned
to Agouron Pharmaceutical describes antiproliferative substituted
quinazolinones and their counterparts wherein the keto group is
replaced by a sulfone.
[0010] U.S. Pat. No. 5,719,157 to Takeda Chemical Industries
describes pharmaceutical compositions for inhibiting bone
resorption which include 4-phenyl quinoline derivatives which may
further be substituted at the 2-position with an optionally
substituted hydrocarbon group or an optionally substituted
heterocyclic group.
[0011] None of the foregoing patents describes quinazoline
derivatives which specifically inhibit p38-.alpha. or
TGF-.beta..
DISCLOSURE OF THE INVENTION
[0012] The invention is directed to methods and compounds useful in
treating conditions that are characterized by enhanced p38-.alpha.
activity and/or TGF-.beta. activity. These conditions include
inflammation, proliferative diseases, and certain cardiovascular
disorders as further described below.
[0013] Compounds of the invention have been found to inhibit p38
kinase, the .alpha.-isoform in particular, and/or TGF-.beta. and
are thus useful in treating diseases mediated by these activities.
The compounds of the invention are of the formula 2
[0014] or the pharmaceutically acceptable salts thereof
[0015] wherein R.sup.3 is a noninterfering substituent;
[0016] each Z is CR.sup.2 or N, wherein no more than two Z
positions in ring A are N, and wherein two adjacent Z positions in
ring A cannot be N;
[0017] each R.sup.2 is independently a noninterfering
substituent;
[0018] L is a linker;
[0019] n is 0 or 1; and
[0020] Ar' is the residue of a cyclic aliphatic, cyclic
heteroaliphatic, aromatic or heteroaromatic moiety optionally
substituted with 1-3 noninterfering substituents.
[0021] The invention is directed to methods of treating
inflammation or proliferative conditions using these compounds. The
invention is also directed to treating conditions associated with
cardiac failure using the invention compounds.
BRIEF DESCRIPTION OF THE DRAWINGS
[0022] FIGS. 1A-1C show the structures of compounds prepared
according to the methods of the invention and useful in the
invention methods.
MODES OF CARRYING OUT THE INVENTION
[0023] The compounds of formula (1) are useful in treating
conditions which are characterized by overactivity of p38 kinase,
in particular the .alpha.-isoform and/or overactivity of
TGF-.beta.. Conditions "characterized by enhanced p38-.alpha.
activity" include those where this enzyme is present in increased
amount or wherein the enzyme has been modified to increase its
inherent activity, or both. Conditions "characterized by enhanced
TGF-.beta. activity" include those wherein TGF-.beta. synthesis is
stimulated so that TGF-.beta. is present in enhanced amount or
wherein TGF-.beta. latent protein is undesirably activated or
converted to active TGF-.beta. protein or wherein TGF-.beta.
receptors are upregulated or wherein the TGF-.beta. protein shows
enhanced binding to cells or extracellular matrix in the location
of the disease. Thus, in either case, "enhanced activity" refers to
any condition wherein the effectiveness of either of these proteins
is undesirably high, regardless of the cause.
[0024] The compounds of the invention are useful in conditions
where either p38-.alpha. kinase or TGF-.beta. shows enhanced
activity since these compounds inhibit the activities of both
proteins. This is particularly advantageous in conditions which are
characterized by enhanced activities of both proteins. These
conditions are those in which fibrosis and organ sclerosis are
caused by, or accompanied by, inflammation, oxidation injury,
hypoxia, altered temperature or extracellular osmolarity,
conditions causing cellular stress, apoptosis or necrosis. These
conditions include ischemia-reperfusion injury, congestive heart
failure, progressive pulmonary and bronchial fibrosis, hepatitis,
arthritis, inflammatory bowel disease, glomerular sclerosis,
interstitial renal fibrosis, chronic scarring diseases of the eyes,
bladder and reproductive tract, bone marrow dysplasia, chronic
infectious or autoimmune states and traumatic or surgical wounds.
These conditions, of course, would be benefited by compounds which
inhibit either TGF-.beta. or p38-.alpha.. They are especially
benefited by treatment with compounds that inhibit both. Methods of
treatment with the compounds of the invention are further discussed
below.
[0025] The Invention Compounds
[0026] The compounds useful in the invention are derivatives of
quinazoline and related compounds containing mandatory substituents
at positions corresponding to the 2- and 4-positions of
quinazoline. In general, a quinazoline nucleus is preferred,
although alternatives within the scope of the invention are also
illustrated below. Preferred embodiments for Z.sup.3 are N and CH;
preferred embodiments for Z.sup.5-Z.sup.8 are CR.sup.2. However,
each of Z.sup.5-Z.sup.8 can also be N, with the proviso noted
above. Thus, with respect to the basic quinazoline type ring
system, preferred embodiments include quinazoline per se, and
embodiments wherein all of Z.sup.5 -Z.sup.8 as well as Z.sup.3 are
either N or CH. Also preferred are those embodiments wherein
Z.sup.3 is N, and either Z.sup.5 or Z.sup.8 or both Z.sup.5 and
Z.sup.8 are N and Z.sup.6 and Z.sup.7 are CH or CR.sup.2. Where
R.sup.2 is other than H, it is preferred that CR.sup.2 occur at
positions 6 and/or 7.
[0027] As used herein, a "noninterfering substituent" is a
substituent which leaves the ability of the compound of formula (1)
to inhibit p38-.alpha. activity and/or TGF-.beta. activity
qualitatively intact. Thus, the substituent may alter the degree of
inhibition and the balance between p38-.alpha. inhibition and
TGF-.beta. inhibition. However, as long as the compound of formula
(1) retains the ability to inhibit either p38-.alpha. or TGF-.beta.
activity or both, the substituent will be classified as
"noninterfering."
[0028] As used herein, "hydrocarbyl residue" refers to a residue
which contains only carbon and hydrogen. The residue may be
aliphatic or aromatic, straight-chain, cyclic, branched, saturated
or unsaturated. The hydrocarbyl residue, when indicated, may
contain heteroatoms over and above the carbon and hydrogen members
of the substituent residue. Thus, when specifically noted as
containing such heteroatoms, the hydrocarbyl residue may also
contain carbonyl groups, amino groups, hydroxyl groups and the
like, or contain heteroatoms within the "backbone" of the
hydrocarbyl residue.
[0029] As used herein, the term "alkyl," "alkenyl" and "alknyl"
include straight- and branched-chain and cyclic monovalent
substituents. Examples include methyl, ethyl, isobutyl, cyclohexyl,
cyclopentylethyl, 2-propenyl, 3-butynyl, and the like. Typically,
the alkyl, alkenyl and alkynyl substituents contain 1-10C (alkyl)
or 2-10C (alkenyl or alkynyl). Preferably they contain 1-6C (alkyl)
or 2-6C (alkenyl or alkynyl). Heteroalkyl, heteroalkenyl and
heteroalkynyl are similarly defined but may contain 1-2 O, S or N
heteroatoms or combinations thereof within the backbone
residue.
[0030] As used herein, "acyl" encompasses the definitions of alkyl,
alkenyl, alkynyl and the related hetero-forms which are coupled to
an additional residue through a carbonyl group.
[0031] "Aromatic" moiety refers to a monocyclic or fused bicyclic
moiety such as phenyl or naphthyl; "heteroaromatic" also refers to
monocyclic or fused bicyclic ring systems containing one ore more
heteroatoms selected from O, S and N. The inclusion of a heteroatom
permits inclusion of 5-membered rings as well as 6-membered rings.
Thus, typical aromatic systems include pyridyl, pyrimidyl, indolyl,
benzimidazolyl, benzotriazolyl, isoquinolyl, quinolyl,
benzothiazolyl, benzofuranyl, thienyl, furyl, pyrrolyl, thiazolyl,
oxazolyl, imidazolyl and the like. Any monocyclic or fused ring
bicyclic system which has the characteristics of aromaticity in
terms of electron distribution throughout the ring system is
included in this definition. Typically, the ring systems contain
5-12 ring member atoms.
[0032] Similarly, "arylalkyl" and "heteroalkyl" refer to aromatic
and heteroaromatic systems which are coupled to another residue
through a carbon chain, including substituted or unsubstituted,
saturated or unsaturated, carbon chains, typically of 1-6C. These
carbon chains may also include a carbonyl group, thus making them
able to provide substituents as an acyl moiety.
[0033] With respect to the substituent at the positions
corresponding to the 4-position of quinazoline, LAr', L is present
or absent and is a linker which spaces the substituent Ar' from
ring B at a distance of 2-8 .ANG., preferably 2-6 .ANG., more
preferably 2-4 .ANG.. The distance is measured from the ring carbon
in ring B to which one valence of L is attached to the atom of the
Ar' cyclic moiety to which the other valence of the linker is
attached. The Ar' moiety may also be coupled directly to ring B
(i.e., when n is 0). Typical, but nonlimiting, embodiments of L are
of the formula S(CR.sup.2.sub.2).sub.m,
--NR.sup.1SO.sub.2(CR.sup.2.s- ub.2).sub.l,
NR.sup.1(CR.sup.2.sub.2).sub.m, NR.sup.1CO(CR.sup.2.sub.2).su- b.l,
O(CR.sup.2.sub.2).sub.m, OCO(CR.sup.2.sub.2).sub.l, and 3
[0034] wherein Z is N or CH and wherein m is 0-4 and l is 0-3,
preferably 1-3 and 1-2, respectively. L preferably provides
--NR.sup.1-- coupled directly to ring B. A preferred embodiment of
R.sup.1 is H, but R.sup.1 may also be acyl, alkyl, arylacyl or
arylalkyl where the aryl moiety may be substituted by 1-3 groups
such as alkyl, alkenyl, alkynyl, acyl, aryl, alkylaryl, aroyl,
N-aryl, NH-alkylaryl, NH-aroyl, halo, OR, NR.sub.2, SR, --SOR,
--NRSOR, --NRSO.sub.2R, --SO.sub.2R, --OCOR, --NRCOR,
--NRCONR.sub.2, --NRCOOR, --OCONR.sub.2, --RCO, --COOR,
--SO.sub.3R, --CONR.sub.2, SO.sub.2NR.sub.2, CN, CF.sub.3, and
NO.sub.2, wherein each R is independently H or alkyl (1-4C),
preferably the substituents are alkyl (1-6C), OR, SR or NR.sub.2
wherein R is H or lower alkyl (1-4C). More preferably, R.sup.1 is H
or alkyl (1-6C). Any aryl groups contained in the substituents may
further be substituted by for example alkyl, alkenyl, alkynyl,
halo, OR, NR.sub.2, SR, --SOR, --SO.sub.2R, --OCOR, --NRCOR,
--NRCONR.sub.2, --NRCOOR, --OCONR.sub.2, --RCO, --COOR, SO.sub.2R,
NRSOR, NRSO.sub.2R, --SO.sub.3R, --CONR.sub.2, SO.sub.2NR.sub.2,
CN, CF.sub.3, or NO.sub.2, wherein each R is independently H or
alkyl (1-4C).
[0035] Ar' is aryl, heteroaryl, including 6-5 fused heteroaryl,
cycloaliphatic or cycloheteroaliphatic. Preferably Ar' is phenyl,
2-, 3- or 4-pyridyl, indolyl, 2- or 4-pyrimidyl, benzimidazolyl,
indolyl, preferably each optionally substituted with a group
selected from the group consisting of optionally substituted alkyl,
alkenyl, alkynyl, aryl, N-aryl, NH-aroyl, halo, OR, NR.sub.2, SR,
--OOCR, --NROCR, RCO, --COOR, --CONR.sub.2, SO.sub.2NR.sub.2, CN,
CF.sub.3, and NO.sub.2, wherein each R is independently H or alkyl
(1-4C).
[0036] Ar' is more preferably indolyl, 6-pyrimidyl, 3- or
4-pyridyl, or optionally substituted phenyl.
[0037] For embodiments wherein Ar' is optionally substituted
phenyl, substituents include, without limitation, alkyl, alkenyl,
alkynyl, aryl, alkylaryl, aroyl, N-aryl, NH-alkylaryl, NH-aroyl,
halo, OR, NR.sub.2, SR, --SOR, --SO.sub.2R, --OCOR, --NRCOR,
--NRCONR.sub.2, --NRCOOR, --OCONR.sub.2, RCO, --COOR, --SO.sub.3R,
--CONR.sub.2, SO.sub.2NR.sub.2, CN, CF.sub.3, and NO.sub.2, wherein
each R is independently H or alkyl (1-4C). Preferred substituents
include halo, OR, SR, and NR.sub.2 wherein R is H or methyl or
ethyl. These substituents may occupy all five positions of the
phenyl ring, preferably 1-2 positions, preferably one position.
Embodiments of Ar' include substituted or unsubstituted phenyl, 2-,
3-, or 4-pyridyl, 2-, 4- or 6-pyrimidyl, indolyl, isoquinolyl,
quinolyl, benzimidazolyl, benzotriazolyl, benzothiazolyl,
benzofuranyl, pyridyl, thienyl, furyl, pyrrolyl, thiazolyl,
oxazolyl, imidazolyl, and morpholinyl. Particularly preferred as an
embodiment of Ar' is 3- or 4-pyridyl, especially 4-pyridyl in
unsubstituted form.
[0038] Any of the aryl moieties, especially the phenyl moieties,
may also comprise two substituents which, when taken together, form
a 5-7 membered carbocyclic or heterocyclic aliphatic ring.
[0039] Thus, preferred embodiments of the substituents at the
position of ring B corresponding to 4-position of the quinazoline
include 2-(4-pyridyl)ethylamino; 4-pyridylamino; 3-pyridylamino;
2-pyridylamino; 4-indolylamino; 5-indolylamino; 3-methoxyanilinyl;
2-(2,5-difluorophenyl)ethylamino-, and the like.
[0040] R.sup.3 is generally a hydrocarbyl residue (1-20C)
containing 0-5 heteroatoms selected from O, S and N. Preferably
R.sup.3 is alkyl, aryl, arylalkyl, heteroalkyl, heteroaryl, or
heteroarylalkyl, each unsubstituted or substituted with 1-3
substituents. The substituents are independently selected from a
group that includes halo, OR, NR.sub.2, SR, --SOR, --SO.sub.2R,
--OCOR, --NRCOR, --NRCONR.sub.2, --NRCOOR, --OCONR.sub.2, RCO,
--COOR, --SO.sub.3R, NRSOR, NRSO.sub.2R, --CONR.sub.2,
SO.sub.2NR.sub.2, CN, CF.sub.3, and NO.sub.2, wherein each R is
independently H or alkyl (1-4C) and with respect to any aryl or
heteroaryl moiety, said group further including alkyl (1-6C) or
alkenyl or alkynyl. Preferred embodiments of R.sup.3 (the
substituent at position corresponding to the 2-position of the
quinazoline) comprise a phenyl moiety optionally substituted with
1-2 substituents preferably halo, alkyl (1-6C), OR, NR.sub.2, and
SR wherein R is as defined above. Thus, preferred substituents at
the 2-position of the quinazoline include phenyl, 2-bromophenyl,
2-chlorophenyl, 2-fluorophenyl, 2-methylphenyl, 4-fluorophenyl and
the like. Other preferred embodiments of R.sup.3 comprise a
cyclopentyl or cyclohexyl moiety.
[0041] As noted above, R.sup.2 is a noninterfering substituent. As
set forth above, a "noninterfering substituent" is one whose
presence does not substantially destroy the p38-.alpha. kinase
inhibiting ability and/or TGF-.beta. inhibiting ability of the
compound of formula (1).
[0042] Each R.sup.2 is also independently a hydrocarbyl residue
(1-20C) containing 0-5 heteroatoms selected from O, S and N.
Preferably, R.sup.2 is independently H, alkyl, alkenyl, alkynyl,
acyl or hetero-forms thereof or is aryl, arylalkyl, heteroalkyl,
heteroaryl, or heteroarylalkyl, each unsubstituted or substituted
with 1-3 substituents selected independently from the group
consisting of alkyl, alkenyl, alkynyl, aryl, alkylaryl, aroyl,
N-aryl, NH-alkylaryl, NH-aroyl, halo, OR, NR.sub.2, SR, --SOR,
--SO.sub.2R, --OCOR, --NRCOR, --NRCONR.sub.2, --NRCOOR, NRSOR,
NRSO.sub.2R, --OCONR.sub.2, RCO, --COOR, --SO.sub.3R, NRSOR,
NRSO.sub.2R, --CONR.sub.2, SO.sub.2NR.sub.2, CN, CF.sub.3, and
NO.sub.2, wherein each R is independently H or alkyl (1-4C). The
aryl or aroyl groups on said substituents may be further
substituted by, for example, alkyl, alkenyl, alkynyl, halo, OR,
NR.sub.2, SR, --SOR, --SO.sub.2R, --OCOR, --NRCOR, --NRCONR.sub.2,
--NRCOOR, --OCONR.sub.2, RCO, --COOR, --SO.sub.3R, --CONR.sub.2,
SO.sub.2NR.sub.2, CN, CF.sub.3, and NO.sub.2, wherein each R is
independently H or alkyl (1-4C). More preferably the substituents
on R.sup.2 are selected from R.sup.4, halo, OR.sup.4,
NR.sup.4.sub.2, SR.sup.4, --OOCR.sup.4, --NR.sup.4OCR.sup.4,
--COOR.sup.4, R.sup.4CO, --CONR.sup.4.sub.2,
--SO.sub.2NR.sup.4.sub.2, CN, CF.sub.3, and NO.sub.2, wherein each
R.sup.4 is independently H, or optionally substituted alkyl (1-6C),
or optionally substituted arylalkyl (7-12C) and wherein two R.sup.4
or two substituents on said alkyl or arylalkyl taken together may
form a fused aliphatic ring of 5-7 members.
[0043] R.sub.2 may also, itself, be selected from the group
consisting of halo, OR, NR.sub.2, SR, --SOR, --SO.sub.2R, --OCOR,
--NRCOR, --NRCONR.sub.2, --NRCOOR, NRSOR, NRSO.sub.2R,
--OCONR.sub.2, RCO, --COOR, --SO.sub.3R, NRSOR, NRSO.sub.2R,
--CONR.sub.2, SO.sub.2NR.sub.2, CN, CF.sub.3, and NO.sub.2, wherein
each R is independently H or alkyl (1-4C).
[0044] More preferred substituents represented by R.sup.2 are those
as set forth with regard to the phenyl moieties contained in Ar' or
R.sup.3 as set forth above. Two adjacent CR.sup.2 taken together
may form a carbocyclic or heterocyclic fused aliphatic ring of 5-7
atoms. Preferred R.sup.2 substituents are of the formula R.sup.4,
--OR.sup.4, SR.sup.4 or R.sup.4NH--, especially R.sup.4NH--,
wherein R.sup.4 is defined as above. Particularly preferred are
instances wherein R.sup.4 is substituted arylalkyl.
[0045] The compounds of formula (1) may be supplied in the form of
their pharmaceutically acceptable acid-addition salts including
salts of inorganic acids such as hydrochloric, sulfuric,
hydrobromic, or phosphoric acid or salts of organic acids such as
acetic, tartaric, succinic, benzoic, salicylic, and the like. If a
carboxyl moiety is present on the compound of formula (1), the
compound may also be supplied as a salt with a pharmaceutically
acceptable cation.
[0046] Synthesis of the Invention Compounds
[0047] The compounds of the invention may be synthesized from the
corresponding 4-halo-2-phenyl quinazoline as described in Reaction
Scheme 1; which may be obtained from the corresponding
4-hydroxyquinazoline as shown in Reaction Scheme 2. Alternatively,
the compounds can be prepared using anthranylamide as a starting
material and benzoylating the amino group followed by cyclization
to obtain the intermediate 2-phenyl-4-hydroxy quinazoline as shown
in Reaction Scheme 3. Reaction Schemes 4-6 are similar to Reaction
Scheme 3 except that an appropriate pyridine or 1,4-pyrimidine
nucleus, substituted with a carboxamide residue and an adjacent
amino residue, is substituted for the anthranylimide. The compounds
of the invention wherein R.sup.1 is H can be further derivatized to
comprise other embodiments of R.sup.1 as shown in Reaction Scheme
7. 4
[0048] Reaction Scheme 1 is illustrative of the simple conversion
of a halogenated quinazoline to compounds of the invention. Of
course, the phenyl of the illustration at position 2 may be
generalized as R.sup.3 and the 4-pyridylamino at position 2 can be
generalized to Ar'--L or Ar'--. 5
[0049] Reaction Scheme 2 can, of course, be generalized in the same
manner as set forth for Reaction Scheme 1. 67
[0050] Again, Reaction Scheme 3 can be generalized by substituting
the corresponding acyl halide, R.sup.3COCl for the
parafluorobenzoyl chloride. Further, Ar' or Ar'--L may be
substituted for 4-aminopyridine in the last step. 8 9 10
[0051] It is seen that Reaction Scheme 1 represents the last step
of Reaction Schemes 2-6 and that Reaction Scheme 2 represents the
last two steps of Reaction Scheme 3-6.
[0052] Reaction Scheme 7 provides conditions wherein compounds of
formula (1) are obtained wherein R.sup.1 is other than H. 11
[0053] Reaction Scheme 8 is a modification of Reaction Scheme 3
which simply demonstrates that substituents on ring A are carried
through the synthesis process. The principles of the behavior of
the substituents apply as well to Reactions Schemes 4-6. 1213
[0054] Reaction Scheme 8 shows a modified form of Reaction Scheme 3
which includes substituents R.sup.2 in the quinazoline ring of
formula (1). The substituents are carried throughout the reaction
scheme. In step a, the starting material is treated with thionyl
chloride in the presence of methanol and refluxed for 12 hours. In
step b, the appropriate substituted benzoyl chloride is reacted
with the product of step a by treating with the appropriately
substituted benzoyl chloride in pyridine for 24 hours. In
embodiments wherein X (shown illustratively in the ortho-position)
is fluoro, 2-fluorobenzoyl chloride is used as a reagent; where X
is (for illustration ortho-chloro), 2-chlorobenzoyl chloride is
used.
[0055] In step c, the ester is converted to the amide by treating
in ammonium hydroxide in an aprotic solvent such as dimethyl
formamide (DMF) for 24 hours. The product is then cyclized in step
d by treatment with 10 N NaOH in ethanol and refluxed for 3
hours.
[0056] The resulting cyclized form is then converted to the
chloride in step e by treating with thionyl chloride in chloroform
in the presence of a catalytic amount of DMF under reflux for 4
hours. Finally, the illustrated 4-pyridylamino compound is obtained
in step f by treating with 4-amino pyridine in the presence of
potassium carbonate and DMF and refluxed for 2 hours.
[0057] In illustrative embodiments of Reaction Scheme 8, R.sup.2
may, for example, provide two methoxy substituents so that the
starting material is 2-amino-4,5-dimethoxy benzoic acid and the
product is, for example,
2-(2-chlorophenyl)-4-(4-pyridylamino)-6,7-dimethoxyquinazoline.
[0058] In another illustrative embodiment, R.sup.2 provides a
single nitro; the starting material is thus, for example,
2-amino-5-nitrobenzoic acid and the resulting compound is, for
example, 2(2-fluorophenyl)-4-(4-p-
yridylamino)-5-nitroquinazoline.
[0059] Reaction Schemes 4-6 can be carried out in a manner similar
to that set forth in Reaction Scheme 8, thus carrying along R.sup.2
substituents through the steps of the process.
[0060] In compounds of the invention wherein R.sup.2 is nitro, the
nitro group may be reduced to amino and further derivatized as
indicated in Reaction Scheme 9. 14
[0061] In Reaction Scheme 9, the illustrative product of Reaction
Scheme 8 is first reduced in step g by treating with hydrogen and
palladium on carbon (10%) in the presence of acetic acid and
methanol at atmospheric pressure for 12 hours to obtain the amino
compound. The resulting amino compound is either converted to the
acyl form (R=acyl) using the appropriate acid chloride in the
presence of chloroform and pyridine for four hours, or is converted
to the corresponding alkylated amine (R=alkyl) by treating the
amine intermediate with the appropriate aldehyde in the presence of
ethanol, acetic acid, and sodium triacetoxyborohydride for 4
hours.
[0062] While the foregoing exemplary Reaction Schemes are set forth
to illustrate the synthetic methods of the invention, it is
understood that the substituents shown on the quinazoline ring of
the products are generically of the formula (1) as described herein
and that the reactants may be substituted accordingly. Variations
to accommodate various substituents which represent embodiments of
R.sup.3 other than the moieties shown in these illustrative
examples or as Ar' in these illustrative examples may also be used.
Similarly, embodiments wherein the substituent at position 4
contains an arylalkyl can be used in these schemes. Methods to
synthesize the compounds of the invention are, in general, known in
the art.
[0063] Administration and Use
[0064] The compounds of the invention are useful among other
indications in treating conditions associated with inflammation.
Thus, the compounds of formula (1) or their pharmaceutically
acceptable salts are used in the manufacture of a medicament for
prophylactic or therapeutic treatment of mammals, including humans,
in respect of conditions characterized by excessive production of
cytokines and/or inappropriate or unregulated cytokine activity on
such cells as cardiomyocytes, cardiofibroblasts and
macrophages.
[0065] The compounds of the invention inhibit the production of
cytokines such as TNF, IL-1, IL-6 and IL-8, cytokines that are
important proinflammatory constituents in many different disease
states and syndromes. Thus, inhibition of these cytokines has
benefit in controlling and mitigating many diseases. The compounds
of the invention are shown herein to inhibit a member of the MAP
kinase family variously called p38 MAPK (or p38), CSBP, or SAPK-2.
The activation of this protein has been shown to accompany
exacerbation of the diseases in response to stress caused, for
example, by treatment with lipopolysaccharides or cytokines such as
TNF and IL-1. Inhibition of p38 activity, therefore, is predictive
of the ability of a medicament to provide a beneficial effect in
treating diseases such as coronary artery disease, congestive heart
failure, cardiomyopathy, myocarditis, vasculitis, restenosis, such
as occurs following coronary angioplasty, atherosclerosis,
rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis, gouty
arthritis and other arthritic conditions, multiple sclerosis, acute
respiratory distress syndrome (ARDS), asthma, chronic obstructive
pulmonary disease (COPD), silicosis, pulmonary sarcosis, sepsis,
septic shock, endotoxic shock, toxic shock syndrome, heart and
brain failure (stroke) that are characterized by ischemia and
reperfusion injury, surgical procedures, such as transplantation
procedures and graft rejections, cardiopulmonary bypass, coronary
artery bypass graft, CNS injuries, including open and closed head
trauma, inflammatory eye conditions such as conjunctivitis and
uveitis, acute renal failure, glomerulonephritis, inflammatory
bowel diseases, such as Crohn's disease or ulcerative colitis,
graft vs host disease, bone resorption diseases like osteoporosis,
type II diabetes, pyresis, psoriasis, cachexia, viral diseases such
as those caused by HIV, CMV, and Herpes, and cerebral malaria.
[0066] Within the last several years, p38 has been shown to
comprise a group of MAP kinases designated p38-.alpha., p38-.beta.,
p38-.gamma. and p38-.delta.. Jiang, Y. et al. J Biol Chem
(1996)271:17920-17926 reported characterization of p38-.beta. as a
372-amino acid protein closely related to p38-.alpha.. In comparing
the activity of p38-.alpha. with that of p38-.beta., the authors
state that while both are activated by proinflammatory cytokines
and environmental stress, p38-.beta. was preferentially activated
by MAP kinase kinase-6 (MKK6) and preferentially activated
transcription factor 2, thus suggesting that separate mechanisms
for action may be associated with these forms.
[0067] Kumar, S. et al. Biochem Biophys Res Comm (1997) 235:533-538
and Stein, B. et al. J Biol Chem (1997) 272:19509-19517 reported a
second isoform of p38-.beta., p38-.beta.2, containing 364 amino
acids with 73% identity to p38-.alpha.. All of these reports show
evidence that p38-.beta. is activated by proinflammatory cytokines
and environmental stress, although the second reported p38-.beta.
isoform, p38-.beta.2, appears to be preferentially expressed in the
CNS, heart and skeletal muscle compared to the more ubiquitous
tissue expression of p38-.alpha.. Furthermore, activated
transcription factor-2 (ATF-2) was observed to be a better
substrate for p38-.beta.2 than for p38-.alpha., thus suggesting
that separate mechanisms of action may be associated with these
forms. The physiological role of p38-.beta.1 has been called into
question by the latter two reports since it cannot be found in
human tissue and does not exhibit appreciable kinase activity with
the substrates of p38-.alpha..
[0068] The identification of p38-.gamma. was reported by Li, Z. et
al. Biochem Biophys Res Comm (1996) 228:334-340 and of p38-.delta.
by Wang, X., et al., J Biol Chem (1997) 272:23668-23674 and by
Kumar, S., et al., Biochem Biophys Res Comm (1997) 235:533-538. The
data suggest that these two p38 isoforms (.gamma. and .delta.)
represent a unique subset of the MAPK family based on their tissue
expression patterns, substrate utilization, response to direct and
indirect stimuli, and susceptibility to kinase inhibitors.
[0069] Various results with regard to response to drugs targeting
the p38 family as between p38-.alpha. and either the putative
p38-.beta.1 or p38-.beta.2 or both were reported by Jiang, Kumar,
and Stein cited above as well as by Eyers, P. A. et al. Chem and
Biol (1995) 5:321-328. An additional paper by Wang, Y. et al. J
Biol Chem (1998) 273:2161-2168 suggests the significance of such
differential effects. As pointed out by Wang, a number of stimuli,
such as myocardial infarction, hypertension, valvular diseases,
viral myocarditis, and dilated cardiomyopathy lead to an increase
in cardiac workload and elevated mechanical stress on
cardiomyocytes. These are said to lead to an adaptive hypertrophic
response which, if not controlled, has decidedly negative
consequences. Wang cites previous studies which have shown that in
ischemia reperfusion treated hearts, p38 MAPK activities are
elevated in association with hypertrophy and programmed cell death.
Wang shows in the cited paper that activation of p38-.beta.
activity results in hypertrophy, whereas activation of p38-.alpha.
activity leads to myocyte apoptosis. Thus, selective inhibition of
p38-.alpha. activity as compared to p38-.beta. activity will be of
benefit in treating conditions associated with cardiac failure.
These conditions include congestive heart failure, cardiomyopathy,
myocarditis, vasculitis, vascular restenosis, valvular disease,
conditions associated with cardiopulmonary bypass, coronary artery
bypass, grafts and vascular grafts. Further, to the extent that the
.alpha.-isoform is toxic in other muscle cell types,
.alpha.-selective inhibitors would be useful for conditions
associated with cachexia attributed to TNF or other conditions such
as cancer, infection, or autoimmune disease.
[0070] Thus, the invention encompasses the use of compounds which
selectively inhibit the activity of the p38-.alpha. isoform for
treating conditions associated with activation of p38-.alpha., in
particular those associated with cardiac hypertrophy, ischemia or
other environmental stress such as oxidation injury,
hyperosmolarity or other agents or factors that activate
p38-.alpha. kinase, or cardiac failure, for example, congestive
heart failure, cardiomyopathy and myocarditis.
[0071] The TGF-.beta. inhibition activity is useful in treating
fibroproliferative diseases, treating collagen vascular disorders,
treating eye diseases associated with a fibroproliferative
condition, venting excessive scarring, treating neurological
conditions and other conditions that are targets for TGF-.beta.
inhibitors and in preventing excessive scarring that elicits and
accompanies restenosis following coronary angioplasty, cardiac
fibrosis occurring after infarction and progressive heart failure,
and in hypertensive vasculopathy, and keloid formation or
hypertrophic scars occurring during the healing of wounds including
surgical wounds and traumatic lacerations.
[0072] Neurological conditions characterized by TGF-.beta.
production include CNS injury after traumatic and hypoxic insults,
Alzheimer's disease, and Parkinson's disease.
[0073] Other conditions that are potential clinical targets for
TGF-.beta. inhibitors include myelofibrosis, tissue thickening
resulting from radiation treatment, nasal polyposis, polyp surgery,
liver cirrhosis, and osteoporosis.
[0074] Diseases benefited by TGF-.beta. inhibition include
cardiovascular diseases such as congestive heart failure, dilated
cardiomyopathy, myocarditis, or vascular stenosis associated with
atherosclerosis, angioplasty treatment, or surgical incisions or
mechanical trauma; kidney diseases associated with fibrosis and/or
sclerosis, including glomerulonephritis of all etiologies, diabetic
nephropathy, and all causes of renal interstitial fibrosis,
including hypertension, complications of drug exposure, such as
cyclosporin, HIV-associated nephropathy, transplant nephropathy,
chronic ureteral obstruction; hepatic diseases associated with
excessive scarring and progressive sclerosis, including cirrhosis
due to all etiologies, disorders of the biliary tree, and hepatic
dysfunction attributable to infections such as hepatitis virus or
parasites; syndromes associated with pulmonary fibrosis with
consequential loss of gas exchange or ability to efficiently move
air into and out of the lungs, including adult respiratory distress
syndrome, idiopathic pulmonary fibrosis, or pulmonary fibrosis due
to infectious or toxic agents such as smoke, chemicals, allergens,
or autoimmune disease; all collagen vascular disorders of a chronic
or persistent nature including progressive systemic sclerosis,
polymyositis, scleroderma, dermatomyositis, fascists, or Raynaud's
syndrome, or arthritic conditions such as rheumatoid arthritis; eye
diseases associated with fibroproliferative states, including
proliferative vitreoretinopathy of any etiology or fibrosis
associated with ocular surgery such as retinal reattachment,
cataract extraction, or drainage procedures of any kind; excessive
or hypertrophic scar formation in the dermis occurring during wound
healing resulting from trauma or surgical wounds; disorders of the
gastrointestinal tract associated with chronic inflammation, such
as Crohn's disease or ulcerative colitis or adhesion formation as a
result of trauma or surgical wounds, polyposis or states post polyp
surgery; chronic scarring of the peritoneum associated with
endometriosis, ovarian disease, peritoneal dialysis, or surgical
wounds; neurological conditions characterized by TGF-.beta.
production or enhanced sensitivity to TGF-.beta., including states
post-traumatic or hypoxic injury, Alzheimer's disease, and
Parkinson's disease; and diseases of the joints involving scarring
sufficient to impede mobility or produce pain, including states
post-mechanical or surgical trauma, osteoarthritis and rheumatoid
arthritis.
[0075] The modulation of the immune and inflammation systems by
TGF-.beta. (Wahl et al. Immunol Today (1989) 10:258-61) includes
stimulation of leukocyte recruitment, cytokine production, and
lymphocyte effector function, and inhibition of T-cell subset
proliferation, B-cell proliferation, antibody formation, and
monocytic respiratory burst. TGF-.beta. is a stimulator for the
excess production of extracellular matrix proteins, including
fibronectin and collagen. It also inhibits the production of
enzymes that degrade these matrix proteins. The net effect is the
accumulation of fibrous tissue which is the hallmark of
fibroproliferative diseases.
[0076] TGF-.beta. is active as a homodimer, but is synthesized and
secreted from cells as an inactive latent complex of the mature
homodimer and proregions, called latency associated protein (LAP).
These proteins bind to each other through noncovalent interactions
(Lyons and Moses Eur J Biochem (1990) 187:467). LAP is often
disulfide-linked to separate gene products, called latent
TGF-.beta. binding proteins or LTBPs. These latent forms provide
stability for the mature cytokine and a means for targeting it to
the extracellular matrix and cell surfaces (Lawrence Eur Cytokine
Network (1996) 7:363-74). Activation of the latent complex occurs
after secretion from cells and is believed to result from the
action of proteases, such as plasmin (Munger et al. Kidney Intl
(1997) 51:1376-82), on LAP, thrombospondin-1 binding (Crawford et
al. Cell (1998) 93:1159-70), and binding to the integrin v6 (Munger
et al. Cell (1999) 319-28).
[0077] Other than v6 there is a variety of cell surface
proteins/receptors that transduce the signals initiated by binding
of the active TGF-.beta. ligand to its receptors. These include
types I, II, III, IV, and V. Type IV is present only in the
pituitary gland while the others are ubiquitous. The binding
affinities among the three isoforms for the type I and II receptors
differ such that these two receptors bind TGF-.beta.1 and
TGF-.beta.3 [?] more tightly than TGF-.beta.2 (Massague Cell (1992)
69:1067-70).
[0078] The type IV receptor or endoglin has a similar isoform
binding profile in contrast to the type III receptor, betaglycan,
which binds equally well to all three isoforms (Wang et al. Cell
(1991) 67:797-805; Lopez-Casillas Cell (1991) 67:785-95). The type
V receptor binds to IGFBP-3 and is thought to have an active kinase
domain similar to the type I and II receptors. Cloning of the type
I and type II receptors demonstrated the existence of cytoplasmic
serine/threonine kinase domains (Wrana et al. Cell (1992)
71:1003-14; Lin et al. Cell (1992) 68:775-85; Ibid. 71:1069;
Massague Cell (1992) 69:1067-70). Initiation of the TGF-.beta.
signaling pathway results from the binding of the TGF-.beta. ligand
to the extracellular domain of the type II receptor (Massague Ann
Rev Biochem (1998) 67:753-91). The bound receptor then recruits
type I receptor into a multimeric membrane complex, whereupon the
constitutively active type II receptor kinase phosphorylates and
activates type I receptor kinase. The function of the type I
receptor kinase is to phosphorylate a receptor-associated
co-transcription factor, smad-2/3, thereby releasing it into the
cytoplasm where it binds to smad-4. This smad complex translocates
into the nucleus, associates with a DNA-binding cofactor, such as
Fast-1, binds to enhancer regions of specific genes, and activates
transcription. The expression of these genes leads to the synthesis
of cell cycle regulators that control proliferative responses or
extracellular matrix proteins that mediate outside-in cell
signaling, cell adhesion, migration, and intercellular
communication.
[0079] The manner of administration and formulation of the
compounds useful in the invention and their related compounds will
depend on the nature of the condition, the severity of the
condition, the particular subject to be treated, and the judgement
of the practitioner; formulation will depend on mode of
administration. As the compounds of the invention are small
molecules, they are conveniently administered by oral
administration by compounding them with suitable pharmaceutical
excipients so as to provide tablets, capsules, syrups, and the
like. Suitable formulations for oral administration may also
include minor components such as buffers, flavoring agents and the
like. Typically, the amount of active ingredient in the
formulations will be in the range of 5%-95% of the total
formulation, but wide variation is permitted depending on the
carrier. Suitable carriers include sucrose, pectin, magnesium
stearate, lactose, peanut oil, olive oil, water, and the like.
[0080] The compounds useful in the invention may also be
administered through suppositories or other transmucosal vehicles.
Typically,.such formulations will include excipients that
facilitate the passage of the compound through the mucosa such as
pharmaceutically acceptable detergents.
[0081] The compounds may also be administered topically, for
topical conditions such as psoriasis, or in formulation intended to
penetrate the skin. These include lotions, creams, ointments and
the like which can be formulated by known methods.
[0082] The compounds may also be administered by injection,
including intravenous, intramuscular, subcutaneous or
intraperitoneal injection. Typical formulations for such use are
liquid formulations in isotonic vehicles such as Hank's solution or
Ringer's solution.
[0083] Alternative formulations include nasal sprays, liposomal
formulations, slow-release formulations, and the like, as are known
in the art.
[0084] Any suitable formulation may be used. A compendium of
art-known formulations is found in Remington's Pharmaceutical
Sciences, latest edition, Mack Publishing Company, Easton, Pa.
Reference to this manual is routine in the art.
[0085] The dosages of the compounds of the invention will depend on
a number of factors which will vary from patient to patient.
However, it is believed that generally, the daily oral dosage will
utilize 0.001-100 mg/kg total body weight, preferably from 0.01-50
mg/kg and more preferably about 0.01 mg/kg-10 mg/kg. The dose
regimen will vary, however, depending on the conditions being
treated and the judgment of the practitioner.
[0086] It should be noted that the compounds of formula (1) can be
administered as individual active ingredients, or as mixtures of
several embodiments of this formula. In addition, the inhibitors of
p38 kinase or TGF-.beta., and dual inhibitors of p38kinase and
TGF-.beta. kinase, can be used as single therapeutic agents or in
combination with other therapeutic agents. Drugs that could be
usefully combined with these compounds include natural or synthetic
corticosteroids, particularly prednisone and its derivatives,
monoclonal antibodies targeting cells of the immune system,
antibodies or soluble receptors or receptor fusion proteins
targeting immune or non-immune cytokines, and small molecule
inhibitors of cell division, protein synthesis, or mRNA
transcription or translation, or inhibitors of immune cell
differentiation or activation.
[0087] As implicated above, although the compounds of the invention
may be used in humans, they are also available for veterinary use
in treating animal subjects.
[0088] The following examples are intended to illustrate but not to
limit the invention.
EXAMPLE 1
Synthesis of 4-(4-pyridylamino)-2-penyl quinazoline
[0089] This example illustrates Reaction Scheme 1.
[0090] A. 4-Chloro-2-phenyl quinazoline, 1 equivalent, was treated
with 1 equivalent 4-aminopyridine and 1 equivalent potassium
carbonate in dimethylformamide (DMF), under reflux for 4 hours. The
reaction mixture was cooled to room temperature and concentrated
under vacuum to an oil. This crude material was dissolved in ethyl
acetate and chromatographed using hexane:ethyl acetate:methanol
8:2:0.5 to obtain solid product. Electron impact mass spectroscopy
(EIMS) gave a molecular ion corresponding to the calculated
molecular weight of the title compound.
[0091] B. Using the procedure of paragraph A of the example but
substituting the starting materials shown in Table 1 below for
4-aminopyridine, the corresponding quinazolines shown in the table
were obtained.
1TABLE 1 Substitute for 4-amino pyridine Product obtained 3-amino
pyridine 2-phenyl-4-(3-pyridylami- no)-quinazoline 2-amino pyridine
2-phenyl-4-(2-pyridylamino)-quinaz- oline 4-aminomethyl
2-phenyl-4-(2-(4-pyridyl)methylamino)-quinazoli- ne pyridine
3-aminomethyl 2-phenyl-4-(2-(3-pyridyl)methylami- no)-quinazoline
pyridine 2-aminomethyl
2-phenyl-4-(2-(2-pyridyl)methylamino)-quinazoline pyridine
EXAMPLE 2
Synthesis of 4-(4-pyridylamino)-2-(4-chlorophenyl) quinazoline
[0092] This example illustrates Reaction Scheme 2.
[0093] A. 4-Chloro-2-(4-chlorophenyl) quinazoline:
4-hydroxy-2-(4-chloroph- enyl) quinazoline, 1 equivalent, was
suspended in chloroform and treated with 12 equivalents of thionyl
chloride in the presence of a catalytic amount of dimethyl
formamide, under reflux for 4 hours. After removal of the solvents
under reduced pressure, a solid was obtained that was analyzed by
thin layer chromatography and EIMS and found to be
4chloro-2-(4-chlorophenyl) quinazoline.
[0094] B. 4-(4-pyridylamino)-2-(4-chlorophenyl) quinazoline:
4-chloro-2-(4-chlorophenyl) quinazoline, 1 equivalent, was treated
with 1 equivalent 4-aminopyridine and 1 equivalent potassium
carbonate in dimethylformamide (DMF), under reflux for 4 hours, as
described in Example 1. The reaction mixture was worked up as in
Example 1 and product confirmed by EIMS.
EXAMPLE 3
Synthesis of 4-(4-pyridylamino)-2-(4-fluorophenyl) quinazoline
[0095] This example illustrates Reaction Scheme 3.
[0096] A. 4-Fluorobenzoyl anthranilamide: Anthranilamide, 1
equivalent, was dissolved in chloroform/pyridine (1:1) and treated
with 4-fluorobenzoyl chloride, 1.1 equivalent for one hour at room
temperature. The reaction was concentrated under vacuum. The
residue was taken up in ethyl acetate and washed with 1 N aqueous
sodium carbonate, 10% aqueous hydrochloric acid, saturated sodium
chloride solution and dried over anhydrous sodium sulfate.
Concentration of the ethyl acetate layer gave a white solid that
was found to be homogenous by thin layer chromatography (TLC) and
confirmed by EIMS.
[0097] B. 4-Hydroxy-2-(4-fluorophenyl) quinazoline: 4-fluorobenzoyl
anthranilamide, from paragraph A, 1 equivalent, was dissolved in
ethanol and to this was added 10 N aqueous sodium hydroxide, 3.0
equivalents, and the resulting solution heated under reflux for 3
hours. The reaction mixture was cooled to room temperature and
concentrated under vacuum. The residue was dissolved in an excess
of water and acidified with concentrated hydrochloric acid. A white
precipitate forms upon acidification. This precipitate was filtered
and washed extensively with water. The solid was then dried under
high vacuum in the presence of dessicant. The solid was found to be
homogenous by TLC and product confirmed by EIMS.
[0098] C. 4-Chloro-2-(4-fluorophenyl) quinazoline:
4-hydroxy-2-(4-fluoroph- enyl) quinazoline, from paragraph B, 1
equivalent, was suspended in chloroform and treated with 12
equivalents of thionyl chloride in the presence of a catalytic
amount of dimethyl formamide, under reflux for 4 hours. After
removal of the solvents under reduced pressure a solid was obtained
that was analyzed by TLC. EIMS confirmed the desired product.
[0099] D. 4-(4-pyridylamino)-2-(4-fluorophenyl) quinazoline:
4-chloro-2-(4-fluorophenyl) quinazoline from paragraph C was
reacted as in Example 1 to obtain the title compound.
EXAMPLE 4
Synthesis of 2-Phenyl-4-(3-methoxyaniliniyl) quinazoline
[0100] 4-Chloro-2-phenylquinazoline, 2 equivalents,
3-methoxyanilinyl, 2 equivalents, and potassium carbonate, 2
equivalents, were dissolved in 10 mL isopropanol and refluxed for 2
hours. The precipitated product formed was filtered and washed with
water. Recrystallization from methanol provided the product as a
white solid that was found to be homogenous by thin layer
chromatography (TLC) and confirmed by EIMS.
EXAMPLE 5
Synthesis of 4-(4-Methoxybenzyl-4-pyridylamino)-2-phenyl
quinazoline
[0101] 4-(4-pyridylamino)-2-phenyl quinazoline, 1 equivalent, was
dissolved in reagent grade acetone, to this was added 5 equivalents
of potassium hydroxide and 1.5 equivalents of 4-methoxybenzyl
chloride. The mixture was refluxed under nitrogen for 4 hours.
After cooling to room temperature the reaction mixture was
concentrated and the residue taken up in ethyl acetate and washed
with saturated aqueous sodium chloride and dried over anhydrous
sodium sulfate and concentrated to give an oil. This crude material
was dissolved in ethyl acetate and chromatographed as in Example 1.
EIMS confirmed the product.
EXAMPLE 6
Prepared Compounds of the Invention
[0102] The compounds in Table 2 shown below have been prepared
using the reaction schemes and exemplary procedures set forth
herein. In the compounds of Table 2, Z.sup.5-Z.sup.8 are CH and
Z.sup.3 is N; i.e., these are all quinazoline derivatives per se.
The table thus lists the embodiments of L, Ar and R.sup.3.
2TABLE 2 Compound No. L Ar' R.sup.3 1 NH 4-pyridyl 2-chlorophenyl 2
NH 4-pyridyl 2,6-dichlorophenyl 3 NH 4-pyridyl 2-methylphenyl 4 NH
4-pyridyl 2-bromophenyl 5 NH 4-pyridyl 2-fluorophenyl 6 NH
4-pyridyl 2,6-difluorophenyl 7 NH 4-pyridyl phenyl 8 NH 4-pyridyl
4-fluorophenyl 9 NH 4-pyridyl 4-methoxyphenyl 10 NH 4-pyridyl
3-fluorophenyl 11* N* 4-pyridyl phenyl .sup. 12.sup..dagger.
N.sup..dagger. 4-pyridyl phenyl 13 NHCH.sub.2 4-pyridyl phenyl 14
NHCH.sub.2 4-pyridyl 4-chlorophenyl 15 NH 3-pyridyl phenyl 16
NHCH.sub.2 2-pyridyl phenyl 17 NHCH.sub.2 3-pyridyl phenyl 18
NHCH.sub.2 2-pyridyl phenyl 19 NHCH.sub.2CH.sub.2 2-pyridyl phenyl
20 NH 6-pyrimidinyl phenyl 21 NH 2-pyrimidinyl phenyl 22 NH phenyl
phenyl 23 NHCH.sub.2 phenyl 3-chlorophenyl 24 NH 3-hydroxyphenyl
phenyl 25 NH 2-hydroxyphenyl phenyl 26 NH 4-hydroxyphenyl phenyl 27
NH 4-indolyl phenyl 28 NH 5-indolyl phenyl 29 NH 4-methoxyphenyl
phenyl 30 NH 3-methoxyphenyl phenyl 31 NH 2-methoxyphenyl phenyl 32
NH 4-(2-hydroxyethyl)phenyl phenyl 33 NH 3-cyanophenyl phenyl 34
NHCH.sub.2 2,5-difluorophenyl phenyl 35 NH 4-(2-butyl)phenyl phenyl
36 NHCH.sub.2 4-dimethylaminophenyl phenyl 37 NH 4-pyridyl
cyctopentyl 38 NH 2-pyridyl phenyl 39 NHCH.sub.2 3-pyridyl phenyl
40 NH 4-pyrimidyl phenyl .sup. 41.sup..dagger-dbl.
N.sup..dagger-dbl. 4-pyridyl phenyl 42 NH p-aminomethylphenyl
phenyl 43 NHCH.sub.2 4-aminophenyl phenyl 44 NH 4-pyridyl
3-chlorophenyl 45 NH phenyl 4-pyridyl 46 NH 15 phenyl 47 NH
4-pyridyl t-butyl 48 NH 2-benzylamino-3-pyridyl phenyl 49 NH
2-benzylamino-4-pyridyl phenyl 50 NH 3-benzyloxyphenyl phenyl 51 NH
4-pyridyl 3-aminophenyl 52 NH 4-pyridyl 4-pyridyl 53 NH 4-pyridyl
2-naphthyl 54 16 4-pyridyl phenyl 55 17 phenyl phenyl 56 18
2-pyridyl phenyl 57 NHCH.sub.2CH.sub.2 19 phenyl 58 not present 20
phenyl 59 not present 21 phenyl 60 NH 4-pyridyl cyclopropyl 61 NH
4-pyridyl 2-trifluoromethyl phenyl 62 NH 4-aminophenyl phenyl 63 NH
4-pyridyl cyclohexyl 64 NH 3-methoxyphenyl 2-fluorophenyl 65 NH
4-methoxyphenyl 2-fluorophenyl 66 NH 4-pyrimidinyl 2-fluorophenyl
67 NH 3-amino-4-pyridyl phenyl 68 NH 4-pyridyl 2-benzylaminophenyl
69 NH 2-benzylaminophenyl phenyl 70 NH 2-benzylaminophenyl
4-cyanophenyl 71 NH 3'-cyano-2-benzylaminophenyl phenyl *R.sup.1 =
2-propyl .sup..dagger.R.sup.1 = 4-methoxyphenyl
.sup..dagger-dbl.R.sup.1 = 4-methoxybenzyl
[0103] Compounds 1-37 in Table 2 have been assessed in terms of
percent inhibition of p38-.alpha. activity in the presence of 15
.mu.M concentration (See Example 7). The percent inhibition for all
of these compounds is measurable, and is, for some compounds, as
high as 100%.
[0104] The compounds in Table 3 contain modifications of the
quinazoline nucleus as shown. These compounds have been prepared
and tested for their ability to inhibit TGF-.beta. and/or
p38-.alpha. kinase. All of the compounds in Table 3 are embodiments
of formula (1) wherein Z.sup.3 is N and Z.sup.6 and Z.sup.7
represent CH. In all cases the linker, L, is present and is NH.
3TABLE 3 Compound No. Z.sup.5 Z.sup.8 Ar' R.sup.3 72 CH N 4-pyridyl
2-fluorophenyl 73 CH N 4-pyridyl 2-chlorophenyl 74 CH N 4-pyridyl
phenyl 75 N N 4-pyridyl phenyl 76 N CH 4-pyridyl phenyl
[0105] Additional compounds were prepared wherein ring A contains
CR.sup.2 at Z.sup.6 or Z.sup.7 where R.sup.2 is not H. These
compounds, which are all quinazoline derivatives, wherein L is NH
and Ar' is 4-pyridyl, are shown in Table 4. In Table 4, the percent
inhibition was measured at 15 .mu.M compounds (or at 1 .mu.M
compound as indicated). See Example 7. Inhibitions above 90% were
observed.
4TABLE 4 Compound No. R.sup.3 CR.sup.2 as noted 77 2-chlorophenyl
6,7-dimethoxy 78 2-fluorophenyl 6-nitro 79 2-fluorophenyl 6-amino
80** 2-fluorophenyl 7-amino 81** 2-fluorophenyl
6-(3-methoxybenzylamino) 82** 2-fluorophenyl
6-(4-methoxybenzylamino) 83 2-fluorophenyl 6-(2-isobutylamino) 84
2-fluorophenyl 6-(4-methylmercaptobenzylamino) 85 2-fluorophenyl
6-(4-methoxybenzoyl amino) 86 4-fluorophenyl 7-amino 87
4-fluorophenyl 7-(3-methoxybenzylamino) **Tested at 1 .mu.M
EXAMPLE 7
Assay for p38 Kinase Inhibition
[0106] The compounds to be tested were solubilized in DMSO and
diluted into water to the desired concentrations. The p38 kinase
was diluted to 10 .mu.g/ml into a buffer containing 20 mM MOPS, pH
7.0, 25 mM beta-glycerol phosphate, 2 mg/ml gelatin, 0.5 mM EGTA,
and 4 mM DTT.
[0107] The reaction was carried out by mixing 20 .mu.l test
compound with 10 .mu.l of a substrate cocktail containing 500
.mu.g/ml peptide substrate and 0.2 mM ATP (+200 .mu.Ci/ml
gamma-32P-ATP) in a 4.times. assay buffer. The reaction was
initiated by the addition of 10 .mu.l of p38 kinase. Final assay
conditions were 25 mM MOPS, pH 7.0, 26.25 mM beta-glycerol
phosphate, 80 mM KCl, 22 mM MgCl.sub.2, 3 mM MgSO.sub.4, 1 mg/ml
gelatin, 0.625 mM EGTA, 1 mM DTT, 125 .mu.g/ml peptide substrate,
50 .mu.M ATP, and 2.5 .mu.g/ml enzyme. After a 40 minute incubation
at room temperature, the reaction was stopped by the addition of 10
.mu.l per reaction of 0.25 M phosphoric acid.
[0108] A portion of the reaction was spotted onto a disk of P81
phosphocellulose paper, the filters were dried for 2 minutes and
then washed 4.times. in 75 mM H.sub.3PO.sub.4. The filters were
rinsed briefly in 95% ethanol, dried, then placed in scintillation
vials with liquid scintillation cocktail.
[0109] Alternatively, the substrate is previously biotinylated and
the resulting reactions are spotted on SAM.sup.2TM streptavidin
filter squares (Promega). The filters are washed 4.times. in 2M
NaCl, 4.times. in 2M NaCl with 1% phosphoric acid, 2.times. in
water, and briefly in 95% ethanol. The filter squares are dried and
placed in scintillation vials with liquid scintillation
cocktail.
[0110] Counts incorporated are determined on a scintillation
counter. Relative enzyme activity is calculated by subtracting
background counts (counts measured in the absence of enzyme) from
each result, and comparing the resulting counts to those obtained
in the absence of inhibitor.
[0111] IC.sub.50 values were determined with curve-fitting plots
available with common software packages. Approximate IC.sub.50
values were calculated using formula
IC.sub.50 (app)=A.times.i/(1-A)
[0112] where A=fractional activity and i=total inhibitor
concentration.
[0113] The compounds in Table 5 have IC.sub.50 in the range of
0.1-1.5 .mu.M vs p38-.alpha.:
5TABLE 5 Compound No. Compound Name 16
2-phenyl-4-(4-pyridylmethylamino)-quinazoline 7
2-phenyl-4-(4-pyridylamino)-quinazoline 8 2-(4-fluorophenyl)-(4-py-
ridylamino)-quinazoline 1
2-(2-chlorophenyl)-(4-pyridylamino)-quina- zoline 30
2-phenyl-4-(3-methoxyanilinyl)-quinazoline 5
2-(2-fluorophenyl)-4-(4-pyridylamino)-quinazoline 4
2-(2-bromophenyl)-4-(4-pyridylamino)-quinazoline 3
2-(2-methylphenyl)-4-(4-pyridylamino)-quinazoline 79
2-(2-fluorophenyl)-4-(4-pyridylamino)-6-amino quinazoline
[0114] Compounds 5 and 7 were tested for their specificity for p38
by assessing their ability to inhibit other kinases. These
compounds were tested at 50 .mu.M and were soluble at 250 .mu.M in
5% DMSO/95% water. The results are shown in Table 6.
6TABLE 6 IC.sub.50 (app) - .mu.M DNA-dep Compound p38-y JNK1 PKA
PKC PK (PKD) cck2 EGF-R 5 227 167 >250 >100 120 245 4.2 7
>300 >300 310 >500 240 >500 34
[0115] In Table 6, compound 5 is
2-(2-fluorophenyl)-4-(4-pyridylamino)-qui- nazoline and compound 7
is 2-phenyl-4-(4-pyridylamino)-quinazoline.
[0116] As seen in Table 6, these compounds are highly specific for
p38-.alpha.. In addition, these compounds were assessed with
respect to p38-.beta. and gave curve fitted values of IC.sub.50 as
follows: Compound 5: 0.928 .mu.M; Compound 7: 3.65 .mu.M.
* * * * *