Note: Descriptions are shown in the official language in which they were submitted.
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
CYANOGUANIDINES AND CYANOAMIDINES AS ERBB2 AND EGFR INHIBITORS
Field of the Invention
This invention relates to a series of cyanoguanidine quinazoline and
cyanoamidine
quinazoline derivatives that are useful in the treatment of hyperproliferative
diseases, such as
cancer and inflammation, in mammals. This invention also relates to a method
of using such
compounds in the treatment of hyperproliferative diseases in mammals,
especially humans, and
to pharmaceutical compositions containing such compounds.
Background of the Invention
The type I receptor tyrosine kinase family consists of four closely related
receptors:
EFGR (ErbBl or HERD, ErbB2 (HER2), ErbB3 (HER) and ErbBO (HERO). These are
transmembrane glycoprotein receptors which contain an extracellular Iigand
binding region and,
with the expection of erbB3, an intracellular catalytically active tyrosine
kinase domain. These
receptors transmit extracellular signals through the cytosol to the nucleus.
The extracellular
signal is transmitted by ligand binding to the homomeric receptor, with the
exception of erbB2,
of which a high affinity soluble ligand has yet to be identified. After ligand
binding the type I
receptor tyrosine' kinases either homodimerize or heterodimerize with another
member of the
subfamily of receptors. ErbB2 participates in this process by
heteromerization. In fact, it has
been shown that erbB2 is the preferred heterodimerization partner (Mehelsohn
Oncogene 2000).
Dimerization leads to activation by autophosphorylation of the intracellular
domain. ~ This
autophosphorylation recruits other proteins and leads to a phosphorylation
cascade that transmits
the signal throughout the cell. The type I receptor tyrosine kinase family
signals through the
1
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
ras/raf/MEK/MAPI~ pathway as well as the PI3KJAkt pathway. These signaling
pathways lead
to both cell proliferation and cell survival through inhibition of apoptosis.
Several investigations have demonstrated the role of EGFR and ErbB2 in cancer.
Squamous carcinomas of the head and neck, and lung express high levels of
EGFR. Also,
constitutively active EGFR has been found in gliomas, breast cancer and lung
cancer (Salomon
et al Crit Rev Oncol Hematol 1995, 19, 183-232 - on order 6/4/02). ErbB2
overexpression
occurs in ~30% of all breast cancer. It has been also implicated in other
human cancers
including colon, ovary, bladder, stomach, esophagus, lung, uterus and
prostate. ErbB2
overexpression has also been correlated with poor prognosis in human cancer,
including
metastasis, and early relapse (ref - two Slamon refs from Science and IClapper
review).
The type I tyrosine kinase receptor family has been an active area of anti-
cancer research.
Several inhibitors of the EGFR and the ErbB2 signaling pathway have
demonstrated clinical
efficacy in cancer treatment. Herceptin, a humanized version of anti-ErbB2
monoclonal
antibody, was approved for use in breast cancer in the United States in 1998.
Iressa and Tarceva
are small molecule inhibitors of EGFR that are expected to be launched in
2002. In addition,
several other antibodies and small molecules that target the interruption of
the type I tyrosine
kinase receptor signaling pathways are in clinical and preclinical development
(Ciardiello et al).
Several issued patents and applications have appeared describing quinazoline
based type
I receptor tyrosine kinase inhibitors, including WO 00!44728, WO 01/98277, WO
98!02438, GB
2 345 486 A, WO 96/33980, and references contained therein, which are
incorporated herein by
reference.
2
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Summary of the Invention
This invention provides for cyanoguanidine and cyanoamidine substituted 4-
anilino
quinazolines of formula I, and pharmaceutically acceptable salts and prodrugs
thereof, that are
useful in the treatment of hyperprolifexative diseases. Specifically, the
present invention relates
to compounds of formula I that act as EGFR and/or ErbB2 inhibitors. Also
provided are
formulations containing compounds of formula I and methods of using the
compounds to treat a
patient in need thereof. In addition, there are described processes for
preparing the inhibitory
compounds of formula I.
Accordingly, the present invention refers to compounds of the formula (I):
NHR~
A
iV~ H
(I)
wherein at least one of the positions 6 or 7 of the quinazoline ring must be
substituted by a group
A, and the remaining positions on the quinazoline ring may be optionally
substituted by up to
three R2 groups; wherein
X is N, CH or a C-CN group;
R' is independently an aryl or heteroaryl group, substituted by at least one
one R6 group,
and. optionally substituted by up to -three RS groups, where
RS is cyano, chlorine, fluorine, bromine, lower alkyl, trifluoromethyl,
difluoromethyl,
vitro or OR9;
3
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
R6 is hydrogen, cyano, chlorine, fluorine, bromine, trifluoromethyl,
difluoromethyl,
trifluoromethoxy; vitro, C1-Coo alkyl, Ca-C,o alkenyl, C2-Clo alkynyl, C3-C1o
cycloalkyl, C3-C,o
cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl or
heterocyclylalkyl,
where each C,-Cm alkyl, C2-Clo alkenyl, Ca-Coo alkynyl, C3-C~~ cycloalkyl, C3-
CIo
cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl,
heterocyclylalkyl
portion is optionally substituted with up to five groups independently
selected from oxo,
halogen, cyano, vitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy,
azido, -NR~S02R8, -
S02NR9R~, -C(O)R9, -C(O)ORS, -OC(O)R9, -NR~C(O)OR8, -NR~C(O)R9, -C(O)NR~R9, -
NR~R9,
-NRl°C(O)NR7R9, -ORS, -S(O)RB, -SOaR~3, aryl, arylalkyl, heteroaryl,
heteroarylalkyl,
heterocyclyl, and heterocyclylalkyl, and where
R7 and R~° independently represent hydrogen or Cm alkyl, or R' and
Rl° together with
the atom. to which they are attached form a 4 to 10 membered carbocyclic,
heteroaryl or
heterocyclic ring, each of which is optionally substituted with up to three
groups independently
selected from halogen, cyano, vitro, trifluoromethyi, difluoromethoxy,
trifluoromethoxy, azido,
aryl, heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl and
heterocyclylalkyl;
R8 represents trifluoromethyl, CI-Clo alkyl, C3-Coo cycloalkyl, aryl,
arylalkyl, heteroaryl,
heteroarylalkyl, het~ocyclyl or heterocyclylalkyl, where each alkyl,
cycloalkyl, aryl, heteroaryl
and heterocyclyl portion is optionally substituted with one to five groups
independently selected
from oxo, halogen, cyano, vitro, trifluoromethyl, difluoromethoxy,
trifluoromethoxy, azido, (aryl,
heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl or heterocyclylalkyl;
4
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
R9 represents represents hydrogen, trifluoromethyl, C,-Clo alkyl, (CHZ)"C3-C,o
cycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl or
heterocyclylalkyl, where
each alkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl portion is
optionally substituted with up
to five groups independently selected from oxo, halogen, cyano, nitro,
trifluoromethyl,
difluoromethoxy, trifluoromethoxy, azido, aryl, heteroaryl, arylalkyl,
heteroarylalkyl,
heterocyclyl and heterocyclylalkyl, where n = 0 or 4, or R' and R9 together
with the atom to
which they are attached form a 4 to 10 membered carbocyclic, heteroaryl or
heterocyclic ring,
each of which is optionally substituted with up to three groups independently
selected from
halogen, cyano, nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy,
azido, aryl,
heteroaryl, arylalkyl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl;
RI3 represents trifluoromethyl, difluoromethyl, C~-Clo alkyl, C3-Cyo alkenyl,
C3-Cto
alkynyl, C3-Coo cycloalkyl, C3-Cio cycloalkylalkyl, aryl, arylalkyl,
heteroaryl, heteroarylalkyl,
heterocyclyl or heterocyclylalkyl, where each of the above alkyl, alkenyl,
alkynyl, cycloalkyl,
aryl, heteroaryl and heterocyclyl portion of R13 is optionally substituted
with one to five groups
independently selected from oxo, halogen, cyano, vitro, trifluoromethyl,
difluoromethoxy,
trifluoromethoxy, azido, -NR7SO2R8, -SOaNRSR7, -C(O)RS, -C(O)ORS, -OC(O)RS,
-NR~C(O)ORB, -NR~C(O)RS, -C(O)NR1RS, NR~RS, NRl°C(O)NR~RS, -
NRi°C(NCN)NR~RS, -
ORS, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl and
heterocyclylalkyl; where R7,
R8, RS and R' ° are the same as R7, Rg, RS and R' ° defined
above;
Ra represents hydrogen, halogen, cyano, vitro, trifluoromethyl,
difluoromethyl,
trifluoromethoxy, C~-Cio alkyl, C2-Coo alkenyl, C2-Clo alkynyl, C3-Clo
cycloalkyl, C3-Clo
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroaryiaIkyl, heterocyclyl or
heterocyclylaikyl,
where each alkyl, alkenyl, alkynyl, cycioalkyl, aryl, heteroaryl and
heterocyciyl portion is
optionally substituted with up to five groups independently selected from oxo,
halogen, cyano,
vitro, trifluoromethyl, difluoromethaxy, trifluoromethoxy, azido, -NR'SOaR8, -
SOaNRSR',
-C(O)RS, -C(O)ORS, -OC(O)R9, -NR'C(O)OR8, -NR'C(OCR9, -C(O)NR'R9, -NR'R9,
-NR'°C O NR'RS -NRl°C CN NR'RS -ORS -S(O R'3 -S02Ri3 lalk I,
heteroar 1
( ) ~ ~ ) a ~ ) > > ~'fh ~'y Y Y
heteroarylalkyl, heterocyclyl and heterocyclylalkyl, and where R', R8, RS,
R'° and R13 are the
same as R', Rg, RS, R'° and R~3 as defined above;
A is represented by the following formula (II):
NG~,N
1
L m
D
(II)
wherein
T represents CI-C1° alkyl, C~-C~° alkenyl, C2-C1°
alkynyl, C3-C~° cycloalkyl, C3-C1°
cycloalkylalkyl, aryl, arylalkyl, heteroaryi, heteroarylalkyl, heterocyclyl or
heterocyciylalkyl;
where each alkyl, allcenyl, alkynyl, cycloalkyi, aryl, heteroaryi and
heterocyclyl portion is
optionally substituted with up to five groups independently selected from oxo,
halogen, cyano,
vitro, trifluoromethyi, difluoromethoxy, trifluoromethoxy, azido, -NR'S02Rg, -
S02NR9R',
=C(O)RS, -C(O)ORS, -OC(O)RS, -NR'C(O)OR8, -NR'C(O)RS, -C(O)NR'R9, -NR'R9,
-NRI°C(O)NR'RS, -NR'°C(NCI~NR'R9, -ORS, -S(O)RB, -SOzR~3, aryl,
arylalkyl, heteroaryl,
heteroarylalkyl, heterocyciyl and heterocyclylalkyl, where R', R8, RS,
R1° and R~3 are the same
b
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
as R~, Rg, R9, Rl° and R13 as defined above; T may optionally contain
one or more heteroatoms,
which heteroatoms may be further substituted or oxidized; and m is an integer
from 0 to l;
L is a nitrogen atom or a CR4 group where R4 represents hydrogen,
trifluoromethyl,
difluoromethyl, trifluoromethoxy, C1-Clo alkyl, CZ-Clo alkenyl, Ca-Clo
alkynyl, C3-Clo
cycloalkyl, C3-Coo cycloalkylalkyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl, heterocyclyl or
heterocyclylalkyl, where each alkyl, alkenyl, alkynyl, cycloalkyl, aryl,
heteroaryl and
heterocyclyl portion is optionally substituted with up to five groups
independently selected from
oxo, halogen, cyano, nitro, trifluoromethyl, difluoromethoxy,
trifluoromethoxy, azido,
-NR'S02R8, -SOaNRSR', -C(O)RS, -C(O)ORS, -OC(O)R9, -NR'C(O)OR8, -NR'C(OCR9,
-C(O)NR~R9, -NR'R9, NRioC(O)NR'RS, -NRl°C(NCN)NR'RS, -ORS, -S(O)R~3, -
SOzR~3, aryl,
arylalkyl, heteroaryl, heteroarylalkyl, hetervcyclyl and heterocyclylalkyl;
where R', R8, RS, Rlo
and R13 are the same as R', R$, RS, Rl° and R~3 defined above;
Q is selected from CR3R1IRI~ or NR11812, where R3 is the same as RZ defined
above and
Rll and R12 independently represent hydrogen, trifluoromethyl, difluoromethyl,
trifluoromethoxy, C1-Clo alkyl, CZ-Clo alkenyl, CZ-Clo alkynyI, C3-Clo
cycloalkyl, C3-Clo
cycloalkylalkyl, ~ aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl,
heterocyclylalkyl,
-NR'S02Rs, -SOaNRSR', -C(O)RS, -C(O)ORS, -OC(O)RS, -NR'C(O)OR8, -NR'C(O)R9,
-C(O)NR'R9, -NR'lfS, -NRl°C(O)NR~R9, -ORS, -S(O)R13 or -SO2R13, where
each C1-Clo alkyl,
CZ-Clo alkenyl, C~-Clo alkynyl, C3-Clo cycloalkyl, C3-Clo cycloalkylalkyl,
aryl, arylalkyl,
heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl portion may be
optionally
substituted with up to five groups independently selected from oxo, halogen,
cyano, nitro,
trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, -NR'S02R$, -
SO~NR9R', -C(O)RS,
7
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
-C(O)ORS, -OC(O)RS, -NR'C(0)ORg, -NR'C(O)RS, -C(O)NR'RS, -NR'RS, -
NRl°C(O)NR'R9,
-NRi°C(NCN)NR'RS, -ORS, -S(O)RB, -SOzRl3, aryl, arylalkyl, heteroaryl,
heteroarylalkyl,
heterocyclyl and heterocyclylalkyl; where R', Rg, RS, Rl° and R13 are
the same as R', Rg, RS, Rio
and R13 defined above, provided that (i) when Q is CR3R11Riz not more than one
group among
R3, R' 1 or Rya may be simultaneously connected to C through a heteroatom,
(ii) when Q is
CR3Ri1R~2, R3 may not be cyano or halogen, (iii) when Q is NR1~R12, not more
than one group
between R" and Rla may be connected to N through a heteroatom, and (iv) when L
is CR4, Q is
NRi lRu; and
D represents hydrogen, trifluoromethyl, difluoromethyl, C1-C1° alkyl,
C~,-C~° alkenyl, Ca-
C~° alkynyl, C3-Ci° cycloalkyl, C3-Ci° cycloalkylalkyl,
aryl, arylalkyl, heteroaryl,
heteroarylalkyl, heterocyclyl, heterocyclylalkyl, -SOZNRSR', -C(O)RS, -
C(O)ORS, -OC(O)RS or
-C(O)NR'RS, where each alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl
and heterocyclyl
portion is optionally substituted with up to five groups independently
selected from oxo,
halogen, cyano, nitro, trifluoromethyl, difluoromethoxy, trifluoromethoxy,
azido, -NR'SO2R8, -
SO~NRSR', -C(O)RS, -C(O)ORS, -OC(O)RS, -NR'C(O)ORB, -NR'C(O)RS, -C(O)NR'RS, -
NR'RS,
-NR'°C(O)NR'RS, -NRi°C(NCN)NR'RS, -ORS, -S(O)R~3, -SO~R13, aryl,
arylalkyl, heteroaryl,
heteroarylalkyi, heterocyclyl and heterocyclylalkyl, and where R', R$, RS, Rio
and R~3 are the
same as R', R8, RS, R'° and R13 defined above, provided that when L is
N, (i) D is hydrogen or is
selected so that L binds to a carbon atom or to a S(O); group, where i is an
integer from 1 to 2,
and (ii) if m = 1, T is selected so that L binds to a carbon atom or to a
S(O)S group, where j is an
integer from 1 to 2; or Q and D taken together form a S-11 member ring
containing 0-3
heteroatoms in addition to the nitrogen atoms which are part of the
cyanoguanidine or
8
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
cyanoamidine group, with no direct bonding between any two heteroatoms, except
for a bond
between N to S(O)k, where k is an integer from 1 to 2, the carbon atoms of the
said ring
optionally substituted with up to two groups selected from oxo, halogen,
cyano, vitro,
trifluoromethyl, difluoromethoxy, trifluoromethoxy, azido, aryl, arylalkyl,
heteroaryl,
heteroarylalkyl, heterocyclyl, heterocyclylalkyl, alkyl, alkenyl, alkynyl,
cycloalkyl, -NR7SOaR8,
-SOZNRSR7, -C(O)RS, -C(O)ORS, -OC(O)R9, -NR7C(O)ORB, -NR~C(O)R9, -C(O)NR7RS, -
NR7RS, -NRI°C(O)NR7RS, -NRl°C(NCN)NR7RS,-ORS, -S(O)R~3, and -
SO~R13, where R7, R8, RS,
Rl° and R13 are the same as R', R8, RS, R1° and R'3 above, and
each nitrogen atom of the said
ring may be optionally and independently substituted with an R4 group, where
R4 is the same as
R4 defined above.
Examples of preferred embodiments of Ra include, but are not limited to:
F
CI ~ ~ CI O ~ I CI, Me
O ~ N
F '.~. I ~
> >
CI, Me i ~ C!, Me ~ H, CI, Me
O ~N I~ O ~~. ~'~ O.~\ ~\ N~N F
~ / , , i
'''~ N , ''~.~. ,
~/
N
F
9.
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
In another aspect of the invention there is provided a method of treating
hyperproliferative disease comprising administering to a mammal a
therapeutically effective
amount of a compound of the invention.
Detailed Description of the Invention
The novel compounds encompassed by the instant invention are those described
by the
general formula I set forth above, including ~ enantiomers, diastereosisomers,
tautomers,
pharmaceutically acceptable salts, and prodrugs thereof.
Except as expressly defined otherwise, the following definition of terms is
employed
throughout this specification.
By "C,-Clo alkyl", "alkyl" and "lower alkyl" in the present invention is meant
straight or
branched chain alkyl groups having 1-10 carbon atoms,~such as, methyl, ethyl,
propyl, isopropyl,
n-butyl, sec-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl,
2-hexyl, 3-hexyl, 3-
methylpentyl, heptyl, octyl, and the like. Preferred alkyl radicals are C~.~
alkyls. More preferred
alkyl radicals are C~_3 alkyls.
By "C~-Cao alkenyl", "lower alkenyl" and "alkenyl" means straight and branched
hydrocarbon radicals having from 2 to 10 carbon atoms and at Least one double
bond and
includes ethenyl, propenyl, 1-but-3-enyl, 1-pent-3-enyl, 1-hex-S-enyl and the
like. The preferred
alkenyls are lower alkenyl having 3-5 carbon atoms.
By "C2-C~o alkynyl", "lower alkynyl" and "alkynyl" means straight and branched
hydrocarbon radicals having from 2 to 10 carbon atoms and at least one triple
bond and includes
ethynyl, propynyl, butynyl, pentyn-2-yl and the like. The preferred alkynyls
are alkynyl having
3-S carbon atoms.
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
By the term "halogen" in the present invention is meant fluorine, bromine,
chlorine, and
iodine.
By "aryl" is meant an aromatic carbocyclic group having a single ring (e.g.,
phenyl),
multiple rings {e.g., biphenyl), or multiple condensed rings in which at least
one is aromatic,
(e:g., 1,2,3,4-tetrahydronaphthyl, naphthyl), which is optionally mono-, di-,
or trisubstituted
with, e.g., halogen, lower alkyl, lower alkoxy, trifluoromethyl, aryl,
heteroaryl, and hydroxy.
By "heteroaryl" is meant one or more aromatic ring systems of S-, 6-, or 7-
membered
rings which includes fused ring systems (at least one of which is aromatic) of
5-10 atoms
containing at least one and up to four heteroatoms selected from nitrogen,
oxygen, or sulfur.
. Examples of heteroaryl groups are pyridinyl, imidazolyl, pyrimidinyl,
pyrazolyl, triazolyl,
pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl,
isothiazolyl, pyrrolyl,
~quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl,
indazolyl,
indolizinyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridinyl,
purinyl, oxadiazolyl,
triazolyl, thiadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl,
benzothiophenyl, benzothiazolyl,
benzoxazolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furopyridinyl.
Spiro moieties are
also included within the scope of this definition. Heteroaryl groups are
optionally mono-, di-, or
trisubstituted with, e.g., halogen, lower alkyl, lower alkoxy, haloalkyl,
aryl, heteroaryl, and
hydroxy.
As used herein, the terms ~ "carbocycle", "carbocyclyl", "cycloalkyl" or "C3-
C,o
cycloalkyl" refer to saturated carbocyclic radicals having , three to ten
carbon atoms. The
cycloalkyl can be monocyclic, or a polycyclic :fused system, and can be fused
to an aromatic
ring. Examples of such radicals include cyclopropyl, cyclobutyl, cyclopentyl
and cyclohexyl.
11
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
The cycloalkyl groups herein are unsubstituted or, as specified, substituted
in one or more
substitutable positions with various groups. For example, such cycloalkyl
groups may be
optionally substituted with, for example, C~-C6 alkyl, Ct-C6 alkoxy, halogen,
hydroxy, cyano,
nitro, amino, mono(C1-C6)alkylamino, di(Ci-.C6)alkylamino, C2-C6alkenyl, C~-
Cbalkynyl, C1-C6
haloalkyl, C~-Gs haloalkoxy, amino(Cl-C6)alkyl, mono(C1-C6)alkylamino(C~-
Cb)alkyl or di(C1-
C6)alkylamino(C 1-C6)alkyl.
By "heterocycle" or "heterocyclyl" is meant one or more carbocyclic ring
systems of 5-,
6-, or 7-membered rings which includes fused ring systems of 4-10 atoms
containing at least one
and up to four heteroatoms selected from nitrogen, oxygen, or sulfur, and with
the proviso that
the ring of the group does not contain two adjacent 4 or S atoms. A fused
system can be a
heterocycle fused to an aromatic group. Preferred heterocycles include, but
are not limited to,
pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl,
tetrahydropyranyl,
dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino,
thioxanyl,
piperazinyl, homopiperazinyl, azetidinyl, oxetanyl, thietanyl,
homopiperidinyl, oxepanyl,
thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-
pyrrolinyl, 3-
pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, I,3-dioxolanyl,
pyrazolinyl, dithianyl,
dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl,
pyrazolidinylimidazolinyl,
imidazolidinyl, 3-azabicyc~[3.i.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl,
azabicyclo[2.2.2]-
hexanyl, 3H-indolyl and quinolizinyl. Spiro moieties are also included within
the scope of this
definition. The foregoing groups, as derived from the groups listed above, may
be C-attached or
N-attached where such is possible. For instance, a group derived from pyrrole
may be pyrrol-1-
yl (N-attached) or pyrrol-3-yl (C-attached). Further, a group derived from
imidazole may be
12~~
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
imidazol-1-yl (N-attached) or imidazol-3-yl (C-attached). An example of a
heterocyclic group
'wherein 2 ring carbon atoms are substituted with oxo (=O) moieties is l,l-
dioxo-
thiomorpholinyl. The heterocycle groups herein are unsubstituted or, as
specified, substituted in
one or more substitutable positions with various groups. For example, such
heterocycle groups
may be optionally substituted with, for example, C~-C6 alkyl, Ci-C6 alkoxy,
halogen, hydroxy,
cyano, vitro, amino, mono(CI-C6)alkylamino, di(CI-C6)alkylamino, C2-Cbalkenyl,
Ca-C6alkynyl,
C~-C6 haloalkyl, C,-C6 haloalkoxy, amino(C1-C6)alkyl, mono(Ci-C6)alkylamino(C,-
C6)alkyl or
di(C~-C6)alkylamino(CI-C6)alkyl.
The term "arylalkyl" means an alkyl moiety (as defined above) substituted with
one or
more aryl moiety (also as defined above). The preferred aralkyl radicals are
aryl-C,.3_alkyls.
Examples include benzyl, phenylethyl, and the like.
The term "heteroarylalkyl" means an alkyl moiety (as defined above)
substituted with a
heteroaryl moiety (also as defined above). The preferred heteroarylalkyl
radicals, are 5- or 6-
membered heteroaryl-C~_3_alkyl. Examples include oxazolemethyl, pyridylethyl
and the like.
The term "Me" means methyl, "Et" means ethyl, "Bu" means butyl and "Ac" means
acetyl.
The phrase "pharmaceutically acceptable salts)", as used herein, unless
otherwise
indicated, includes salts of acidic and basic groups which may be present in
the compounds of
formula 1 or of the compounds made in accordance with the examples herein. The
compounds
of formula 1 that are basic in nature are capable of forming a wide variety of
salts with various
inorganic and organic acids. The acids that may be used to prepare
pharmaceutically acceptable
acid addition salts of such basic compounds of formula 1 as well as the
compounds prepared in
13
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
the examples are those that form non-toxic acid addition salts, i.e., salts
containing
pharmaceutically acceptable anions, such as the acetate, benzenesulfonate, .
benzoate,
bicarbonate, bisulfate, bitartrate, borate, bromide, calcium, edetate,
camsylate, carbonate,
chloride, clawlanate, citrate, dihydrochloride, edetate, edislyate, estolate,
esylate, ethylsuccinate,
fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate,
hexylresorcinate,~ hydrabamine,
hydrobromide, hydrochloride, iodide, isothionate, lactate, lactobionate,
laurate, malate, rnaleate,
mandelate, mesylate, methylsulfate, mutate, napsylate, nitrate, oleate,
oxalate, pamoate
(embonate), palimitate, pantothenate, phosphate/diphosphate,
polygalacturonate, salicylate,
stearate, subacetate, succinate, tannate,~ tartrate, teoclate, tosylate,
triethiodode, and valerate salts.
Since a single compound of the present invention may include more than one
acidic or basic
moieties, the compounds of the present invention may include mono, di or tri-
salts in a single
compound.
The phrase "pharmaceutically acceptable salts)", as used herein, unless
otherwise
indicated, includes salts of acidic and basic groups which may be present in
the compounds of
formula 1. The compounds of formula I that are basic in nature are capable of
forming a wide
variety of salts with various inorganic and organic acids. The acids that may
be used to prepare
pharmaceutically acceptable acid addition salts of such basic compounds of
formula 1 are those
that form non-toxic acid addition salts, i.e., salts containing
pharmaceutically acceptable anions,
such as the acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate,
bitartrate, borate,
bromide, calcium, edetate, camsylate, carbonate, chloride, clavulanate,
citrate, dihydrochloride,
edetate, edislyate; estolate, esylate, ethylsuccinate, fiunarate, gluceptate,
gluconate, glutamate,
glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide,
hydrochloride, iodide,
14
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate,
mesylate, methylsulfate,
mucate, napsylate, nitrate, oleate, oxalate, pamoate (embonate), palimitate,
pantothenate,
phosphate/diphosphate, polygalacturonate, salicylate, stearate, subacetate,
succinate, tannate,
tartrate, teoclate, tosylate; triethiodode, and valerate salts. Since a single
compound of the
present invention may include more than one acidic or basic moieties, the
compounds of the
present invention may include mono, di or tri-salts in a single compound.
Those compounds of the present invention that are acidic in nature are capable
of forming
basic salts with various pharmaceutically acceptable cations. Examples of such
salts include the
alkali metal or alkaline earth metal salts and, particularly, the calcium,
magnesium, sodium and
potassium salts of the compounds of the present invention.
Certain compounds of formula I may have asymmetric centers and therefore exist
in
different enantiomeric forms. All optical isomers and stereoisomers of the
compounds of formula
. I, and mixtures thereof, are considered to be within the scope of the
invention. With respect to
the compounds of formula I, the invention includes the use of a racemate, one
or more
enantiomeric forms, one or more diastereomeric forms, or mixtures thereof. The
compounds of
formula I may also exist as tautomers. This invention relates to the use of
all such tautomers and
mixtures thereof.
The subject invention also includes isotopically-labeled compounds, which are
identical
to those recited in formula I, but for the fact that one or more atoms are
replaced by an atom
having an atomic mass or mass number different from the atomic mass or mass
number usually
found in nature. Examples of isotopes that can be incorporated into compounds
of the invention
include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur,
fluorine and
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
chloride, such as zH, 3H, 13C, i4C, IsN, is0, 170, 3ip~ 3ap~ 3sS' lsF~ ~d
36C1, respectively.
Compounds of the present invention, prodrugs thereof, and pharmaceutically
acceptable salts of
said, compounds or of said prodrugs which contain the aforementioned isotopes
andlor other
isotopes of other atoms are within the scope of this invention. Certain
isotopically-labeled
compounds of the present invention, for. example those into which radioactive
isotopes such as
3H and 14C are incorporated, are useful in drug andlor substrate tissue
distribution assays.
Tritiated, i.e., 3H and carbon-14, i.e., ~ 14C, isotopes are particularly
preferred fox their ease of
preparation and detectability. Futher, substitution with heavier isotopes such
as deuterium, i.e.,
aH, can afford certain therapeutic advantages resulting from greater metabolic
stability, , for
example increased in vivo half life or reduced dosage requirements and, hence,
may be preferred
in some circumstances. Isotopically labeled compound of formula I of this
invention and
prodrugs thereof can generally be prepared by carrying out procedures
disclosed in the Schemes
and/or in the Examples and Preparations below, by substituting a readily
available isotopically
labeled reagent for a non-isotopically labeled reagent.
This invention also encompasses pharmaceutical compositions containing and
methods of
treating proliferative disorders, or abnormal cell growth, by administering
prodrugs of
compounds of the formula I. Compounds of formula I having free amino, amido,
hydroxy or
carboxylic groups can be converted into prodrugs. Prodrugs include compounds
wherein an
amino acid residue, or a polypeptide chain of two or more (e.g., two, three or
four) amino acid
residues is covalently joined through an amide or ester bond to a free amino,
hydroxy or
carboxylic acid group of compounds of formula I. The amino acid residues
include but are not
limited to the 20 naturally occurring amino acids commonly designated by three
letter symbols
16
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
and also includes 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-
methylhistidine,
norvaline, beta-alanine, gamma-aminobutyric acid, cirtulline, homocysteine,
homoserine,
~omithine and methionine sulfone. Additional types of prodrugs are also
encompassed. For
instance, free carboxyl groups can be derivatized as amides or alkyl esters.
Free hydroxy groups '
may be derivatized using groups including but not limited to hemisuccinates,
phosphate esters,
dimethylaminoacetates, and phosphoryloxymethyloxycarbonyls, as outlined in
Advanced Drug
Delivery Reviews 1996, 19, 115. Carbamate prodrugs of hydroxy and amino groups
are also
included, as are carbonate prodrugs, sulfonate esters and' sulfate esters of
hydroxy groups.
Derivatization of hydroxy groups as (acyloxy)methyl and (acyloxy)ethyl ethers
wherein the acyl
group may be an alkyl ester, optionally substituted with groups including but
not limited to ether,
/amine and carboxylic acid functionalities,. or where the acyl group is an
amino acid ester as
described above, are also encompassed. Prodrugs of this type are described in
J. Med. Chem.
.1996, 39, 10. Free amines can also be derivatized as amides, sulfonamides or
phosphonamides:
All of these prodrug moieties may incorporate groups including but not limited
to ether, amine
and carboxylic acid functionalities.
It is to be understood that in instances where two or more radicals are used
in succession
to define a substituent attached to a structure, the first named radical is
considered to be terminal
and the last named radical is considered to be attached to the structure in
question. Thus, for
example, the radical arylalkyl is attached to the structure in question by the
alkyl group.
The compounds of the invention are administered either singly or in
combination to a
mammal to treat hyperproliferative disease, such as various types of cancer,
e.g., cancer of the
colon, ovary, bladder, stomach, lung, uterus, and prostate. The compound may
be administered
17'
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
via any acceptable route, e.g., infra venous, oral, infra muscular, via
suppository, etc. The
compounds can be formulated as oral dosage forms, e.g., tablets, capsules,
liquid suspension, etc,
as suppositories, or may be_ prepared as a liquid for injection, for example.
The skilled
practitioner can select the appropriate route and dosage amount for treatment
of the specific
hyperproliferative disease to be treated.
The examples below are intended to illustrate embodiments of the invention,
and are not
intended to limit the scope of the specification or claims in any way.
1~
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Example 1
CI
NC. -~ O ~ I F
N
Me2N~N ~ HN
H ~ ~N
~~~J
N
N-(3-(4-[3-Chloro-4-(3-fluoro-benzyloxy)-phenylamino]-quinazolin-6-yl}-prop-2-
ynyl)-N'-
cyano-N",N"-dimethylguanidine
Step A: 6-iodo-4-quinazolinone.
A solution of 2-amino-5-iodobenzoic acid (14.2 g, 50 mmol) and formamidine
acetate
(6.75 g, 65 mmol) in ethanol (204 mL) was refluxed for 20 hours. After cooling
to 0°C the solid
product was collected by filtration. Further drying in a vacuum provided 6-
Iodo-4-quinazolinone
1 I~ g, 81 %) as a gray solid.
Step B: 4-chloro-6-iodoquinazoline.
To a stirred solution of anhydrous dimethyl foramide (DMF) (3.20 ml) in 1,2-
dichloroethane (DCE) (10 ml), cooled in an ice-water bath, is added dropwise
under nitrogen a
solution of oxalyl chloride (5.2 ml, 60 mmol) in DCE (25 ml). A white
precipitate forms during
the addition. After the end of addition the cold bath is removed and the
reaction mixture is
stirred at room temperature for 5 minutes. 6-Iodo-quinazolin-4-of (5.0 g, I8
mmol) is added in
19
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
portions via scoopula under nitrogen flow and the mixture is heated
immediately to reflux.
Heating is continued for 4.5 hours, followed by cooling to room temperature.
The reaction
mixture is poured into excess ice-water mixture (approximately 300 ml) and
extracted with DCM
(approximately 500 ml). The aqueous layer is further extracted with DCM (2x50
ml). T'he
combined organic extracts are dried (Na2S04) and concentrated under reduced
pressure to yield
5.2 g (99%) of desired product as a tan solid.
Step C: 2-chloro-1-(3-fluoro-benzyloxy)-4-vitro-benzene.
Sodium hydride (60% dispersion in oil, 1.4 g, 33.5 mmol) is suspended in dry
DMF (10
ml) under a nitrogen atmosphere and the resulting mixture is cooled in
ice:water. To above
suspension is added dropwise over 15 minutes (3-Fluoro-phenyl)-methanol (2.90
ml, 27 mmol).
Next, to the cold reaction mixture is added dropwise over 20 minutes a
solution of 2-chloro-1-
fluoro-4-vitro-benzene (4.2 g, 24 mmol) in dry DMF (20 ml). Upon the end of
addition the cold
bath is removed and the reaction mixture is stirred for another 4 hours. The
reaction mixture is
poured into 300 ml of ice:water. The resultant solid is isolated by suction
filtration, washed with
water (500 ml), and air dried to yield 5.5 g (20 mmol, 83%) of the clean
desired material as ap
yellow powder.
Step D: 3-chloro-4-(3-fluoro-benzyloxy)-phenylamine.
2-Chloro-1-(3-fluoro-benzyloxy)-4-vitro-benzene (4.08 g, 14.5 mmol) is
suspended in
MeOH (50 ml) and treated wet 5% Ft/C (Degussa type, Aldrich, 1.5 g). The flask
is flushed with
hydrogen gas from a balloon and the reaction mixture is stirred under hydrogen
atmosphere until
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
reaction is judged complete by this-layer chromatography (approximately 2
hours). The reaction
mixture is filtered through a Celite plug and the solvent is removed under
reduced pressure. The
crude product is redissolved in DCM, dried (MgS04) and concentrated to yield
3.1 g (12 mmol,
83°00) of the desired product.
Step E: [3-chloro-4-(3-fluoro-benzyloxy)-phenyl]-(6-iodo-quinazolin-4-yl)-
amine hydrochloride
salt.
3-Chloro-4-(3-fluoro-benzyloxy)-phenylamine (3.1 g, 12 mmol) and 4-chloro-6-
iodo-
quinazoline (3.28 g, I 1.3 mmol) are dissolved in a 1:1 mixture of DCEa-BuOH
(56 ml). The
reaction mixture is refluxed for I9 hours. The product is isolated by suction
filtration through
sintered glass, washed with excess DCM, and air dried to afford 3.8 g
(7.Ommol, 58°f°) of the
clean desired material.
Step F: (3-{4-[3-chloro-4-(3-fluoro-benzyloxy)-phenylamino]-quinazolin-6-yl}-
prop-2-ynyl)-
carbamic acid tert-butyl ester.
A mixture of prop-2-ynyl-carbamic acid tert-butyl ester (978 mg, 6.31 mmol)
and 3-
chloro-4-(3-fluoro-benzyloxy)-phenylamine hydrochloride (3.11 g, 5.74 mmol),
dichlorobis(tri-
phenylphosphine) palladium (II) (210 mg , 0.299 mmol), copper iodide (57 mg,
0.3 mmol), and
diisopropylamine (1.77 mL, 7.28 mmol) in anhydrous THF (40 mL) was 'stirred at
room
temperature for 5 hours. After concentration, the residue was dissolved in
CHzCl2 (50 mL),
washed with aqueous NH4C1 and brine, dried over sodium sulfate, and
concentrated to give the
2I
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
crude product (3.07 g, 100%) as a light yellow solid which was then used
without further
purification.
Step G: [6-(3-amino-prop-1-ynyl)-quinazolin-4-yl]-[3-chloro-4-(3-fluoro-
benzyloxy)-phenyl]-
amore
To a suspension of (3-{4-[3-cliloro-4-(3-fluoro-benzyloxy)-phenylamino]-
quinazolin-6-
yl)-prop-2-ynyl)-carbamic acid tert-butyl ester (2.01 g, 3.78 mmol) in CH2Cla
(3 mL) was added
trifluoroacetic acid (TFA) (3 mL) dropwise. The reaction was stirred at room
temperature for 30
minutes. The reaction mixture was then diluted with CH2C12 (30 mL) and aqueous
saturated
sodium bicarbonate. Phases were separated and the aqueous layer was extracted
with CH2Cla
(30 mL). ~rganic layers were combined, dried over sodium sulfate and
concentrated to give the
crude product ( 1.648 g, 1 O 1 °Ao)as a yellow oil.
Step H: 1-(3-{4-[3-chloro-4-(3-fluoro-benzyloxy)-phenylamino]-quinazolin-6-yl}-
prop-2-ynyl)-
~-phenyl-N-cyano-isourea.
A mixture of [6-(3-amino-prop-I-ynyl)-quinazolin-4-yl]-[3-chloro-4-(3-fluoro-
benzyloxy)-phenyl]-amine (520 mg, I .2 mmol), Biphenyl cyano-carbonimidate (31
S mg, 1.32
mmol), and triethylamine (0.17 mL, 1.2 mmol) in isopropanol (10 mL) was
stirred at room
temperature fox 15 hours. After concentration, the crude white residue (840
mg) was then used
without further purification.
22
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Step I: N-(3-~4-[3-Chloro-4-(3-fluoro-benzyloxy)-phenylamino]-quinazolin-6-yl}-
prop-2-ynyl)-
N'-cyano-N",N"-dimethylguanidine.
A mixture of crude 1-(3-{4-[3-chloro-4-(3-fluoro-benzyloxy)-phenylamino]-
quinazolin-
6-yl}-prop-2-ynyl)-2-phenyl N-cyano-isourea (80 mg, 0.14 mmol), dimethylamine
(0.25 mL,
2M in THF) in isopropanol (3 mL) was heated to 85°C in a sealed tube.
The reaction was cooled
to room temperature after 3 hours. Solvent was removed via rotovap. The
residue was then
purified by FCC to give the final product (35 mg, 47%) as a light yellow
solid. MS ESI (+) m/z
528 (M+1) detected; 1H NMR (400 MHz, deuterated DMSO) 9.9 (s, 1H), 8.? (s,
1H), 8.6 (s,
1H), 8.05 (s, 1H), 7.8 (m, 1H), 7.75 (m, 2H), 7.58 (br, 1H), 7.5 (m, 1H), 7.22-
7.4 (m, 3H), 7.2
(m, 1 H), 5.25 (s, 2H), 4.4 (m, 2H), 3.02 (s, 6H).
Example 2
CI
O
NC~N ~ F
HO~N~N ~ HN
N H ~ ~ ~N
~ ~ NJ
N-(3-{4-[3-Chloro-4-(3-fluoro-benzyloxy)-phenylamino]-quinazolin-6-yl}-prop-2-
ynyl)- N'-
cyano-N"-3-(2-hydroxy-ethyl)guanidine.
23
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
A mixture of crude 1-(3-{4-[3-chloro-4-(3-fluoro-benzyloxy)-phenylamino]-
quinazolin-
6-yl}-prop-2-ynyl)-2-phenyl-N-cyano-isourea (70 mg, 0.12 mmol) (from Step H,
Example 1), 2-
hydroxyethylamine hydrochloride (39 mg, 0.4 mmol) and triethylamine (0.07 mL,
0.5 mmol) in
isopropanol (2 mL) was heated to 85°C in a sealed tube. The reaction
was cooled to room
temperature after 3 hours. Solvent was removed via rotovap. The residue was
then purified by
FCC to give the final product (25 mg, 38%) as a light yellow solid. MS ESI (+)
m/z 544 (M+1)
detected; iH NMR (400 MHz, deuterated DMSO) 9.95 (s, 1H), 8.7 (s, 1H), 8.6 (s,
1H), 8.05 (s,
1 H), 8.0 (s, 1 H), 7.8 (m, 1 H), 7.78 (m, 2H), 7.6 (br, 1 H), 7. S (m, 1 H),
7.22-7.4 (m, 2H), 7.2 (m,
1 H), 7.1 (m, 1 H), 5.25 (s, 2H), 4.25 (m, 2H),-3.5 (m, 2H), 3.25 (m, 2H).
Example 3
Me
NC~N O
I w ~
H2N~N ~ HN N Me
H
~N
J
N
N-cyano-N'-(3-{4-[3-Methyl-4-(6-methyl-pyridin-3-yloxy)-phenylamino]-
quinazolin-6-yl}-
prop-2-ynyl)guanidine
24 .
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Step A: 2-methyl-5-(2-methyl-4-vitro-phenoxy)-pyridine.
To a 500mL flask equipped with addition funnel at 0°C was added NaH
(8.26 g, 95%,
327 mmol), followed by slow addition of DMF (100 mL). The mixture was stirred
for 10
minutes. The addition funnel was then charged with 6-methyl-pyridin-3-of (30.2
g, 277 mmol)
and DMF (100 mL), and the solution in addition funnel was added to the flask
dropwise over 45
minutes. The reaction mixture was stirred at OoC for another 30 minutes once
the addition was
finished. To the addition funnel was then added 4-fluoro-3-methyl-nitrobenzene
(39.1 g, 252
mmol) and DMF ( 100 mL) and the resulting solution was added to the flask
dropwise over 45
minutes. The cold bath was removed at the end of the addition and the reaction
mixture was
allowed to stir at room temperature for 15 hours. The red dark solution was
cooled to 0°C and
water ( 100 mL) was added cautiously to the reaction mixture. The resulting
solution was stirred
for 30 minutes and solid product was purified by filtration and washed with
cold water (500 mL).
The wet solid was dried in vacuo to give product (49.8 g, 8I %).
Step B: 3-methyl-4-(6-methyl-pyridin-3-yloxy)-phenylamine
A mixture of 2-methyl-5-(2-methyl-4-vitro-phenoxy)-pyridine (11.5 g, 47.1
mmol) and
palladium on carbon (300 mg, 10 wt.%, wet) in MeOH (200 mL) was flashed with
hydrogen. A
hydrogen balloon was then applied to the reaction mixture. The reaction was
stinted for 2 hours
and the solution was filtered through a pad of celite, and the pad was washed
with MeOH (300
mL). Concentration of the solution gave crude product (8.8 g, 87%) as light
yellow solid.
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Step C: (6-iodo-quina~olin-4-yl)-[3-methyl-4-(6-methyl-pyridin-3-yloxy)-
phenyl]-amine
hydrochloride
A mixture of 3-methyl-4-(6-methyl-pyridin-3-yloxy)-phenylamine (4.96 g, 23.18
mmol),
4-Chloro-6-iodo-quinazoline 604 g, 22.06 mmol) in tBuOH (60 mL) and DCE (60
mL) was
refluxed for 6 hours. The reaction was cooled to 0°C and the solid
product (8.44 g , 76%) was
isolated by filtration and washed with cold CHaCla (50 mL).
Step'D: (3-{4-[3-Methyl-4-(6-methyl-pyridin-3-yioxy)-phenylamino]-quinazolin-6-
yl}-prop-2-
ynyl)-carbamic acid tert-butyl ester.
A mixture of prop-2-ynyl-carbamic acid tent-butyl ester (2.65 g, 17.08 mmol)
and (6-
Iodo-quinazolin-4-yl)-[3-methyl-4-(6-methyl-pyridin-3-yloxy)-phenyl)-amine
hydrochloride (8.2
g, .16.27 mmol), dichlorobis(triphenylphosphine) palladium (II) (570 mg , 0.81
mmol), copper
iodide (154 mg, 0.81 mmol), and diisopropylamine (4.78 mL, 34.16 mmol) in
anhydrous THF
(80 mL) was stirred at room temperature for 5 hours. After concentration, the
residue was
dissolved in CH2C12 (100 mL), washed with aqueous NH4C1 and brine, dried over
sodium
sulfate, and concentrated to give the crude product (7.89 g, 98%) as a light
yellow solid which
was then used without :further purification.
Step E: [6-(3-Amino-prop-1-ynyl)-quinazolin-4-yl]-[3-methyl-4-(6-methyl-
pyridin-3-yloxy)-
phenyl]-amine.
To a suspension of (3-{4-[3-methyl-4-(6-methyl-pyridin-3-yloxy)-phenylamino)-
quinazolin-6-yl}-prop-2-ynyl)-carbamic acid tert-butyl ester (1.22 g,
2.46.mmo1) in CHaCl2 (3
26
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
mL) was added TFA (3 mL) dropwise. The reaction was stirred at room
temperature for 30
minutes. The reaction mixture was then diluted with CHZCIa (30 mL) and aqueous
saturated
sodium bicarbonate. Phases were separated and the aqueous layer was extracted
with CHZCI2
(30 mL). Organic layers were combined, dried over sodium sulfate and
concentrated to give the
crude product (0.85 g, 88%) as a yellow oil.
Step F: N-cyano-N'-(3~{4-[3-Methyl-4-(6-methyl-pyridin-3-yloxy)-phenylamino]-
quinazoiin-6-
yl }-prop-2-ynyl)guanidine.
A mixture of 2-phenyl-N-cyano-isourea (50 rng, 0.31 mmol) and [6-(3-amino-prop-
1-
ynyl)-quinazolin-4-yl]-[3-methyl-4-(6-methyl-pyridin-3-yloxy)-phenyl]-amine
(30 mg, 0.076
mmol) in isopropanol (3 mL) was heated at 85°C in a sealed tube. The
reaction was cooled to
room temperature after 5 hours. Solvent was removed via rotovap. The residue
was then
purified by FCC to give the final product ( 18 mg, S 1 %) as a light yellow
solid. MS ESI (+) m/z
463 (M+1 ) detected; 1 H NMR (400 MHz, deuterated DMSO) D 9.9 (s, 1 H), 8.7
(s, 1 H), 8.6 (s,
1H), 8.2 (s, 1H), 7.8 (m, 2H), 7.75 (m, 2H), 7.2 (m, 3H), 7.0 (m, 3H), 4.22
(m, 2H), 2.42 (s, 3H),
2:2 (s, 3H). .
27
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Example 4
F
CI
O
CN ~ HN
N~-NH O ~ ~ ~ N
N i NJ
O
N-(5-{4-[3-Chloro-4-(3-fluoro-benzyloxy)-phenylamino]-quinazolin-6-yl}-furan-2-
ylmethyl)-
N'-cyano-morpholine-4-carboxasnidine
Step A: furan-2-ylmethyl-carbamic acid tert-butyl ester.
Furan-2-ylmethylamine (8.0 ml, 91 mmol) and Boc2O (19.8 g, 91 mmol) are
dissolved in
DCM (40 ml) and stirred at room temperature for 1.5 hours. The reaction
mixture is filtered and
concentrated under reduced pressure to afford 17.6 g (85 mmol, 93%) of the
desired product as
. an yellowish solid containing CA 4% t-)3uOH (1H NMR).
Step B: {5-{~-[3-Chloro-4-(3-fluoro-benzyloxy)-phenylamino]-quinazolin-6-yl}-
furan-2-
ylmethyl)-carbamic acid tert-butyl ester
[3-Chloro-4-(3-fluoro-benzyloxy)-phenyl]-(6-iodo-quinazolin-4-yl)-amine
hydrochloride
(0.913 g, 1.68 mmol) (from Step E, Example 1) is dissolved in DMF (20 ml) and
the solution is
degassed under nitrogen. The above solution is added over 10 hours to a heated
(110°C)
degassed suspension of tricyclohexyl phosphine (0.475 g, 1.7 mmol), palladium
dichloride (15.2
28
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
mg, 0.086 mmol), potassium acetate (0.35 g, 3.6 mmol), tetra n-butyl ammonium
bromide (0.552
g, 2.15 mmol), and Furan-2-ylmethyl-carbamic acid tert-butyl ester (2.9 g, 15
mmol) in DMF (5
ml). Heating is continued for 9 hours after the end of addition. The reaction
mixture is cooled,
diluted with water and extracted with EtOAc. The combined organic extracts are
dried (NaaS04)
and concentrated under xeduced pressure. Flash chromatography on silica with
10-50%
EtOAc:hexanes gradient elution yields 0.450 g (0.78 mmol, 46%) of the clean
desired product.
Step C: [6-(5-Aminomethyl-furan-2-yl)-quinazolin-4-yl]-[3-chloro-4-(3-fluoro-
benzyloxy)-
phenyl]-amine. ,
(5-(4-[3-Chloro-4-(3-fluoro-benzyloxy)-phenylamino]-quinazolin-6-yl)-furan-2-
ylmethyl)-carbamic acid tert-butyl ester (0.0218 g, 0.0379 mmol) is dissolved
in DCM (2 ml)
and TFA (2 ml) is added dropwise. The reaction mixture is stirred at room
temperature for 1
hour. The solvent is removed under a nitrogen stream and to the residue are
added consecutively
saturated aqueous potassium carbonate solution and DCM. The resulting mixture
is extracted
with DCM containing 5% THF, the combined organic extracts are dried (Na2S04)
and
concentrated under reduced pressure to yield 17.6 mg (0.037 mmol, 98%) of the
clean desired
product.
Step D: 1-(5-{4-[3-Chloro-4-(3-fluoro-benzyloxy)-phenylamino]-quinazolin-6-yl}-
furan-2-
ylmethyl)-2-phenyl-N-cyano isourea
[6-(S-Aminomethyl-furan-2-yl)-quinazolin-4-yl]-[3-chloro-4-(3-fluoro-
benzyloxy)-
phenyl]-amine ( 148 mg, 0.313 mmol) and diphenyl cyanocarbonimidate (83 mg,
0.348 mmol)
29
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
are suspended in a 1:2 THF:i-PrOH mixture (9 ml), and stirred overnight at
room temperature
under a nitrogen atmosphere. The resulting suspension is used in the next
reaction step without
purification.
Step E: N-(5-{4-[3-Chloro-4-(3-fluoro-benzyloxy)-phenylamino]-quinazolin-6-yl}-
furan-2-
ylmethyl)-N'-cyano-morpholine-4-carboxamidine.
To one third of the crude product suspension from Step H is added morpholine
(0.03 ml,
0:34 mmol) and the reaction mixture is heated for 2 hours at 80-90 °C
in a sealed reaction vial.
The reaction mixture is cooled, morpholine (0.05 ml, 0.57 mmol) is added, and
the heating (80-
90 °C) 'is continued for 1 how. Concentration of the reaction mixture
followed by flash column
chromatography on silica with a 1:3:96 Et3N:MeOH:DCM eluant yields 9.8 mg
(0.016 mmol,
15% yield over steps I and J) of clean desired product. MS ESI (+) m/z 612
(M+1 ) detected; 1 H
NMR (400 MHz, DMSO-d6) 8 8.88 (s, 1H), 8.?6 (s, 1H), 8.56 (s, 1H), 8.18 (d,
1H), 8.03 (s, 1H),
7.90 (s, 1 H), 7.82 (d, 1 H), 7.75 (d, 1 H), 7.47 (m, 1 H), 7.29 (m, 3H), 7.18
(m, 1 H), 7.07 (d, 1 H),
6.55 (d, 1H), 5.27 (s, 2H), 4.62 (d, 2H), 3.63 (m, 2H), 3.51 (m, 2H)
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Example 5
O-~~
CN S ~ HN ~ N
N~N~N \ \ N
HzN,~ ' i NJ
N-cyano-N'-(4-{4-[3-Methyl-4-(6-methyl-pyridin-3-yloxy)-phenylamino]-
quinazolin-6-yl}-
thiazol-2-ylmethyl)guanidine
Step A: (4-Bromo-thiazol-2-yl)-methanol (modified procedure from Nicolaou et
al., Bioorg.
Med. Chem., 7 ( 1999), 665-697)
2,~-Dibromothiazole (4.31 g, 17.7 mmol) are dissolved in anhydrous diethyl
ether (I70
ml) and the solution is cooled to -78 °C (dry ice-acetone bath). n-
Butyllithium (1.6 M in
hexanes, 13 ml, 20.8 mmol) is added dropwise to the reaction mixture and the
resulting solution
is stirred at the same temperature for 30 minutes.. Anhydrous DMF (ml, mmol)
is then added at -
78 °C and, after being stirred at the -78 °C for 30 minutes, the
reaction mixture is warmed to
room temperature over a period of 2~ hours. Hexanes (300 ml), were added and
the resulting
mixture is passed through a short silica cake eluting with 30% EtOAc-hexanes.
The solvents are
evaporated to yield the crude aldehyde which is used directly in the next
step.
To a solution of the above aldehyde in MeOH (80 nil) is added sodium
borohydride (g,
mmol), and the resulting mixture is stirred room temperature for hours.
Hexanes (300 ml) are
added and the mixture is passed through a short silica cake eluting with
EtOAc. The crude
31
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
alcohol is further purified by flash chromatography on silica with 20-50%
EtOAc-hexanes as an
eluant to yield g (mmol, %) of the pure desired product.
Step B: 2-Azidomethyl-4-bromo-thiazole.
(4-Bromo-thiazol-2-yl)-methanol (I.lg, 5.7 mmol) in DMF (15 ml) is treated at
room
temperature under nitrogen atmosphere with trifluoromethanesulfonyl chloride
(0.61 ml, 1
equivalent), and Et3N (0.8 ml, 1 equivalent). The reaction mixture is stirred
for 3 hours at room
temperature before the addition of sodium azide (l.l l g, 3 equivalents),
followed by overnight
stirring at the same temperature. The reaction mixture is diluted with water
and extracted with
DCM and diethyl ether. The combined organic extracts are dried (MgS04) and
concentrated
under reduced pressure to afford the crude product, which is used without
purification in the next
step.
Step C: (4-Bromo-thiazol-2-yl)-methylamine
Crude 2-Azidomethyl-4-bromo-thiazole from Step E is dissolved in a 1:3:2
THF:EtOH:H20 mixture, and treated with Pt02 (wet, approximately 60 mg). The
reaction flask
is flushed with hydrogen from a balloon, and stirring under hydrogen
atmosphere is continued
for 3 hours. The reaction mixture is filtered through a Celite pad, diluted
with DCM and diethyl
ether, and dried (NaZS04). Chromatography on silica pretreated with 1% Et3N in
EtOAc with
EtOAc-MeOH eluant affords 710 mg (3.68 mmol, 65%) of clean desired product.. '
32
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Step D: (4-Bromo-thiazol-2-ylmethyl)-carbamic acid tent-butyl ester
(4-Bromo-thiazol-2-yl)-methylamine (705.3 mg, 3.68 mmol) is dissolved in
anhydrous
DCM (15 ml) and BocaO (898 mg, 4.13 mmol) is added. The reaction mixture is
stirred at room
temperature for 4 hours. Flash chromatography on silica with 0-20% EtOAc-
hexanes affords 770
mg (2.64 mmol, 72%) of pure desired product.
-Step E: (4-Trimethylstannyl- thiazol-2-ylmethyl)-carbamic acid tert-butyl
ester
(4-Bromo-thiazol-2-ylmethyl)-carbamic acid tert-butyl ester (0.46 g, 1.58
mmol) is added
at room temperature to Pd(PPh3)4 (87 mg, 0.075 mmol) in anhydrous toluene (16
ml) under a
nitrogen atmosphere. Hexamethylditin (5.0 g, 15.26 mmol) is added in one
prtion and the
resulting mixture is degassed under nitrogen. The reaction mixture is heated
at 100 °C for 3
hours, then cooled to room temperature and loaded directly on a silica column
pretreated with
1% Et3N in hexanes. Elution with 0-5% EtOAc-hexanes affords the crude product
which is
further purified by flash chromatography on silica with 0-30°f°
EtOAc-hexanes gradient elution
to yield 357.7 mg (0.950 mmol, 60°!°) of clean desired product.
Step F: (4-{4-[3-Methyl-4-(6-methyl-pyridin-3-yloxy)-phenylamino]quinazolin-6-
yl}-thiazol-2-
ylmethyl)-carbamic acid ter-butyl ester
(4-Trimethylstannyl- thiazol-2-ylmethyl)-carbamic acid tert-butyl ester (191
mg, 0.507
mriiol) and [3-Methyl-4-(6-methyl-pyridin-3-yloxy)-phenyl}-(6-iodo-quinazolin-
4-yl)-amine
hydrochloride (0.251 g, 0.478 mmol) {from Step C, Example 3) are dissolved in
anhydrous DMF
under a nitrogen atmosphere. Hunig's base (0.44 ml, 2.53 mmol), and
PdCla(PPh3)2 are added to
33
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
the reaction mixture at room temperature. The reaction mixture is degassed and
heated at 100 °C
overnight. After cooling to room temperature the reaction mixture is diluted
with with water and
thoroughly extracted with EtOAc and, DCM. The combined organic extracts are
dried (NazS04)
and concentrated under reduced pressure. Flash chromatography on silica with
EtOAc-MeOH as
an eluant affords 8I mg (0.14 mmol, 28%) of clean desired product.
Step G: [6-(2-Aminomethyl-thiazol-4-yl)-quinazolin-4-yl]-[3-methyl-4-(6-methyl-
pyridin-3-
yloxy)-phenyl]-amine
(4-{4-[3-Methyl-4-(6-methyl-pyridin-3-yloxy)-phenylamino]quinazolin-6-yl}-
thiazol-2-
ylmethyl)-carbamic acid ter-butyl ester (81 mg, 0.14 mmol) is treated with
concentrated aqueous
hydrochloric acid (0.5 ml) in EtOAc (6 ml). Reaction progress is followed by
LC/MS. Upon
reaction completion saturated aqueous potassium carbonate solution is added ,
the reaction
mixture is diluted with water and thoroughly extracted with DCM and EtOAc. The
combined
organic extracts are dried (Na2SO4) and concentrated under reduced pressure to
yield 45 mg
(0.095 mmol, 68%) of clean desired product.
Step H: 1-(4-{4-[3-Methyl-4-(6-methyl-pyridin-3-yloxy)-phenylamino]quinazolin-
6-yl}-thiazol-
2-ylmethyl)-2-phenyl N-cyano isourea
[6-(2-Aminomethyl-thiazol-4-yl)-quinazolin-4-yl]-[3-methyl-4-(6-methyl-pyridin-
3-
yloxy)-phenyl]-amine (45 mg, 0.095 mmol) is dissolved in a 1:2 i-PrOH:THF
mixture (6 ml).
biphenyl cyanocarbonimidate (28 mg, 0.12 mmol) is added and the reaction
mixture is stirred
overnight at room temperature under a nitrogen atmosphere. To drive the
reaction to completion
34
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
diphenyl cyanocarbonimidate (20 mg, 0.09 mmol) is added to reaction mixture,
which is stirred
at room temperature for another 4 hours. The reaction mixture is then
concentrated and purified
by flash column chromatography on silica eith MeOH-EtOAc as an eluant. The
yield of pure
desired product is 42 mg (0.07 mmol, 74%).
Step I: >~I-cyano-N'-(4-{4-[3-Methyl-4-(b-methyl-pyridin-3-yloxy)-phenylamino]-
quinazolin-6-
yl }-thiazol-2-ylmethyl)guanidine
1-(4-{4-(3-Methyl-4-(6-methyl-p5vridin-3-yloxy)-phenylamino]quinazolin-6-yl}-
thiazol-
2-ylmethyl)-2-phenyl-N-cyano isourea (8.4 mg, 0.014 mmol) is dissolved in a
1:1 mixture of
THF:i-FrOH (2 ml), and treated with 2.0 M ammonia solution in MeOH (0.1 ml).
The reaction
mixture is heated at 80 C in a sealed reaction vial until the reaction is
complete by LCIMS. Flash
chromatography on silica with MeOH-EtOAc as an eluant affords 5.7 mg (0.011
mmol, 79%) of
pure desired product. MS ESI (+) m/z 522 (M+1) detected; 1H NMR (400 MHz, ,
DMSO-d6) 8
9.08 (s, 1 H), 8.57 (s, 1 H), 7.83 (m, 2H), 7.72 (d, 1 H), 7.63 (bs, 1 H),
7.25 (m, 2H), 7.14(bs, 2H),
6.98 (d, 2H), 4.70 (m, 2H), 2.45 (s, 3H), 2.23 (s, 3H).
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Example 6
ci
N
Me H HN
Me2N~N N N~~ I ~ %N
~CN Me0 ~ N
N-(3- { 4-[3-Chloro-4-(pyridin-2-ylmethoxy)-phenylamino]-7-methoxy-quinazolin-
6-yloxy } -
propyl)-N'-cyano-N"-(2-dimethylamino- ethyl)-N"-methylguanidine
Step A: 2-(2-Chloro-4-nitro-phenoxymethyl)-pyridine
Sodium hydride (95%, 0.935 g, 37 mmol) is suspended in dry DMF (20 ml) under a
nitrogen atmosphere and the resulting mixture is cooled in ice water. To above
suspension is
added dropwise over I S minutes pyridin-2-yl-methanol (3.42 g, 31'.3 mmol) in
dry DMF (20
mL). Next, to the cold reaction mixture is added dropwise over 20 minutes a
solution of 2-
Chloro-1-fluoro-4-nitro-benzene (5 g, 28.5 mmol) in dry DMF (20 ml). Upon the
end of
addition the cold bath is removed and the reaction mixture is stirred for
another 36 'hours. Water
(80 mL) was added slowly to the reaction mixture, and a yellow precipitate
resulted. The
resultant solid is isol-ated by suction filtration, washed with water (80 ml),
and air dried to yield
7.52 g (28.5 mmol, 100%) of the clean desired material as a yellow powder.
36
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Step B: 3-Chloro-4-(pyridin-2-ylmethoxy)-phenylamine
2-(2-Chloro-4-nitro-phenoxymethyl)-pyridine (2.4 g, 9.07 mmol) is suspended in
MeOH
(30 ml) and treated wet S% PtIC (Degussa type, Aldrich, 0.8 g). The flask is
flushed with
hydrogen gas from a balloon and the reaction mixture is stirred under hydrogen
atmosphere until
reaction is complete by TLC (ca 2 hours). The reaction mixture is filtered
through a Celite plug
and the solvent is removed under reduced pressure. The crude product is
redissolved in DCM,
dried (MgS04) and concentrated to yield 1.7 g (7.23 mmol, 80%) of the desired
product.
Step C: 2-Amino-5-hydroxy-4-methoxy-benzoic acid
5-Hydroxy-4-methoxy-2-nitro-benzoic acid (18 g, 84.51 mmol, J. Indian Chem.
Soc.
1970, 70, 925) is suspended in MeOH (1 L) and treated PtOa (100 mg). The flask
is flushed with
hydrogen gas and the reaction mixture is stirred under hydrogen atmosphere (45
psi) for 4 hours.
The reaction mixture is filtered through a celite plug and the solvent is
removed under reduced
pressure. The crude product is redissolved in DCM, dried (MgS04) and
concentrated to yield
15.06 g (82.3 mmol, 97%) of the desired product.
Step D: 7-Methoxy-quinazoline-4,6-diol
Piperidine (3 mL, 31 mmol) was added to a mixture of 2-Amino-5-hydroxy-4-
methoxy-
benzoic acid {8.1 g, 44.26 mmol) and triazine (5.38 g, 66.4 mmol) in MeOH (60
mL). The
37
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
reaction was then heated to reflex and stir for 6 hours. The reaction was cool
to OoC. The
product was isolated by filtration and washed with cold MeOH to give 6.37 g
(33.2 mmol, 75%)
of desired product.
Step E: Acetic acid 4-hydroxy-7-methoxy-quinazolin-6-yl ester
A mixture of 7-Methoxy-quinazoline-4,6-diol {6.2 g, 32.3 mmol), Ac20 (100 mL)
and
pyridine (10 mL) was heat to 100°C, and stirred for 3 hours. The
reaction was then cooled to
room temperature and poured to ice water (300 mL). The product was isolated by
filtration,
washed with water (200 mL) and dried to give 7.61 g (32.4 mmol, 100%) of
desired product.
Step F: Acetic acid 4-chloro-7-methoxy-quinazolin-6-yl ester
To a stirred solution of anhydrous DMF (4.5 mL)~in DCE (20 mL), cooled in an
ice-water
bath, is added dropwise under nitrogen a solution of oxalyl chloride (7.9 ml,
90 mmol) in DCE
(40~mL). A white precipitate forms during the addition. After the end of
addition the cold bath is
removed and the reaction mixture is stirred at room temperature for 5 min.
Acetic acid 4-
hydroxy-7-methoxy-quinazolin-6-yl ester (6.5 g, 27.~ mmol) is added in
portions via scoopula
under nitrogen flow and the mixture is heated~immediately to,reflux. Heating
is continued for 3
hours, followed by cooling to room temperature. The reaction mixture is poured
into excess
ice:water mixture (100 mL) and extracted with DCM (500 mL). The aqueous layer
is further
eattracted with DCM (2x50 mL). The combined organic extracts are dried
(Na2S04) and
38
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
concentrated under reduced pressure to yield 5.63 g (22.34 mmol, 80%) of
desired product as a
tan solid.
Step G: 4-[3-Chloro-4-(pyridin-2-ylmethoxy)-phenylamino]-7-methoxy-quinazolin-
6-of
3-Chloro-4-(pyridin-2-ylmethoxy)-phenylamine (3.9 g, 16.62 mmol) and acetic
acid 4-
chloro-7-methoxy-quinazolin-6-yl ester (4.62 g, 18.28 mmol) were dissolved in
a 1:1 mixture of
DCEa-BuOH (50 mL). The reaction mixture was refluxed for 19 hours and then
cooled to room
temp. Solvent was removed via rotovap. The crude residue was then suspended in
MeOH (80
mL) and NH4OH (8 mL, 30% in water) was added to the mixture. Stir for 15 hours
at room
temp. The reaction was heated to 100°C and stirred for 1 hour. Cool to
0°C and product was
isolated by filtration and washed with cold MeOH to give S g (12.2 mmol, 67%
over two steps)
of desired product.
Step H: (3-(4-[3-Chloro-4-(pyridin-2-ylmethoxy)-phenylamino]-7-methoxy-
quinazolin-6-
yloxy}-propyl)-carbamic acid tert-butyl ester
CsOH monohydrate (0.452 g, 2.69 mmol) was added to a mixture of 4-[3-Chloro-4-
(pyridin-2-ylmethoxy)-phenylamino]-7-methoxy-quinazolin-6-of (1 g, 2.45 mmol),
(3-bromo-
propyl)-carbamic acid tert-butyl ester (0.64 g, 2.69 mmol), tetrabutylammonium
iodide (5 mg)
and 4 A molecular sieves (2 g) in DMF (10 mL) at room temp. Stir for 3 hours.
The reaction
mixture was then filtered through celite and washed with EtOAc (30 mL). The
organic solution
39
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
was washed with water (20 mL) and concentrated. FLC (10:1 EtOAc:Hexanes)
provided desired
product (1.02 g, 73.7 %).
Step I: [6-(3-Amino-propoxy)-7-methoxy-quinazolin-4-yl]-[3-chloro-4-(pyridin-2-
ylmethoxy)-
phenyl]-amine
TFA (3 mL) was added drop wise to a suspension of (3-{4-[3-Chloro-4-(pyridin-2-
ylmethoxy)-phenylamino]-7-methoxy-quinazolin-6-yloxy}-propyl)-carbamic acid
tert-butyl ester
(0:9 g, 1.59 mmol) in DCM (3 mL). Stir for 1 hour and the reaction mixture was
diluted with
DCM (20 ml) and sat. NaHCO3 (20 mL). Phases were separated and organic layer
was extracted
with DCM (20 mL). Organic layers were combined, dried (Na2S04), and
concentrated to give
0.7 g (95°fo) of desired product.
Step J: N-(3-{4-[3-ChIoro-4-(pyridin-2-ylmethoxy)-phenylamino]-7-methoxy-
quinazolin-6-
yloxy}-propyl)-N'-cyano-N"-(2-dimethylamino-ethyl)-N"-methylguanidine
The primary amine was functionalized to the corresponding N-cyanoguanidine in
the
similar fashion as described in the previous examples. MS ESI (+) m/z 618
(M+I) detected; ~H
NMR (400 MHz, deuterated DMSO) 8 9.4 (s, I H), 8.6 (d, J=4 Hz, I H), 8.4 (s, 1
H), 8.0-7.8 (m,
4H), 7.7 (dd, J--8,2 Hz, 1 H), 7.6. (d, J--8 Hz, 1 H), 7.3 (t, J --7 Hz, 1 H),
7.2 (d; J 9 Hz, I H), 7.17
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
(s,' 1 H), 5.2 (s, 2 H), 4.18 (t, J--6 Hz, 2H), 3.91 (s, 3H), 3.6-3.4 (m, 4H),
3.39 (t, J=6 Hz, 2H),
3.27 (q, J--6 Hz, 2H), 2.92 (s, 3H), 2.1,(s, 6H), 2.06 (m, 2H).
Example 7
Me H
Me'NYN
N ~CI
N-(3-{4-[3-Chloro-4-(pyridin-2-ylmethoxy)-phenylamino)-7-methoxy-quinazolin-6-
yloxy)-
propyl)-N'-cyano-N",N"-dimethylguanidine
Prepared similarly as N'-(3-{4-[3-Chloro-4-(pyridin-2-ylmethoxy)-phenylamino]-
7-
methoxy-quinazolin-6-yloxy}-propyl)-N-(2-dimethylamino-ethyl)-N-methyl- N-
cyanoguanidine.
MS ESI (+) mlz S61 (M+1) detected;'H NMR (400 MHz, deuterated DMSO) ~ 9.4 (s,
1H), 8.6
(d, J--4 Hz, 1 H), 8.4 (s, 1 H), 7.95 (d, J--3 Hz, 1 H), 7.87 (td, J--8,1 Hz,
1 H), 7.81 (s, 1 H), 7.67
(dd, ~ 8,2 Hz, 1 H), 7.57 (d, J--8 Hz, I H), 7.35 (dd, J--7,5 Hz, 1 H), 7.25
(d, J--9 Hz, 1 H), 7.17 (s,
1H), 7.1 (t, J--6 Hz, 1H), 5.2 (s, 2 H), 4.18 (t, J--6 Hz, 2H), 3.91 (s, 3H),
3.5 (q, J--7 Hz, 2H),
2.94 (s, 6H), 2.1 (m, 2H).
41
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Example 8
CI
O
F
HN
H H
Me'N N NCO I ~ %N
~'CN Me0 ~ N
N-(3-{4-[3-Chloro-4-(3-fluoro-benzyloxy)-phenylamino]-7-methoxy-quinazolin-6-
yloxy} -
propyl)-N'-cyano-N"-methylguanidine
Prepared similarly as N'-(3-{4-[3-Chloro-4-(pyridin-2-ylmethoxy)-phenylamino]-
7-
methoxy-quinazolin-6-yloxy}-propyl)-N-(2-dimethylamino-ethyl)-N-methyl- N-
cyanoguanidine,
except that 3-Chloro-4-(3-fluoro-benzyloxy)-phenylamine in stead of 3-Chloro-4-
(pyridin-2-
ylmethoxy)-phenylamine was used in Step G in Example 6. MS ESI (+) mlz 539
(M+1) '
detected;'H NMR (400 MHz, deuterated I~MSO) 6 9.43 (s, IH), 8.46 (s, IH),
7.96(d, J--3 Hz,
I H), 7.82 (s, 1 H), 7.7 (dd, J--9,2 Hz, 1 H), 7.5 (q, .~ 6 Hz, 1 H), 7.33 (d,
J--8 Hz, 1 H), 7.3 I (d, J--9
Hz, 1 H), 7.27 (d, J--9 Hz, 1.H), 7.2-7. I (m, 2H), 7.03 (m, 1 H), 7.69 (m, 1
H), 5.2 (s, 2 H), 4. I 8 (t,
J--6 Hz, 2H), 3.94 (s, 3H), 2.68 (d, J--6 Hz,,3H), 2.05 (m, 2H).
42
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Example 9
F
N
N .~ I o N
HN
~N H \ ~ ~ N
N
N-cyano-N'-(3-{4-[1-(3-Fluoro-benzyl)-1H-indazol-5-ylamino]-quinazolin-6-yl}-
prop-2-ynyl)-
morpholine-4-carboxamidine
Step A: 1.-(3-Fluoro-benzyl)-5-nitro-1H-indazole
A modified procedure from WO 99/35146, p. 6lwas followed. 5-nitroindazole
(3.915 g,
24 mmol) treated with potassium carbonate (3.65 g, 1.1 equiv.), and 3-
fluorobenzyl bromide (5
g, 1.1 equiv.) in 41 ml of dry DMF under NZ. Reaction mixture is stirred at ?5
°C for 4 hours.
The crude product (yellow solid, 5.536 g) is isolated as in the reference
procedure. Acetone (26
ml) is added to the crude product, and the insoluble solids are filtered off.
To filtered solution is
added water dropwise (12 ml) upon which an oil forms..The mixture is store in
freezer at -20 C
for 15 min, upon which the oil solidifies and remains solid after warming to
r.t. Chromatography
of the solid (silica, 0-10% EtOAclhexanes) afforded 2.49 g of high Rf material
(1-H regioisomer,
9.2 mmol, 38%), 0.7 g of the low Rf material .(2-H isomer, ~11%) and mixed
fractions (0.71 g,
3%).
43
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Step B: 1-(3-Fluoro-benzyl)-1H-indazol-S-ylamine
Follow modified procedure from WO 99/35146. 1-(3-Fluoro-benzyl)-5-vitro-1H-
indazole
(2.49 g, 9.2 mmol) is suspended in 40 ml absolute EtOH and PtIC (5%, wet, 150
mg) is added.
The reaction mixture is stirred and heated at 60 °C under a hydrogen
atmosphere (balloon).
Roughly 4 hours into the reaction LC/MS reveals the formation of substantial
amounts of
product. The mixture is filtered through Celite and concentrated under reduced
pressure. Yield:
2.01 g (90.8%) of a white solid.
Step C: [1-(3-Fluoro-benzyl)-1H-indazol-5-yl]-(6-iodo-quinazolin-4-yl)-amine
hydrochloride
Follow general procedure from Example 1, step E. 4-chloro-6-iodoquinazoline
(1.18 g,
4.06 mmol) is mixed with 1-(3-Fluoro-benzyl)-IH-indazol-5-ylamine (1.09 g,
4.52 mmol), and a
mixture of DCE (lOml) and t-BuQH {IO ml) is added. The mixture is heated at 90
°C (oil bath
temperature) for 8 hours. At 5 hours of heating LC/MS reveals substantial
amount of product.
Yield is 1.35 g (56%).
Step D: (3- f 4-[I-(3-Fluoro-ben~yl)-1H-indazol-5-ylamino]-quinazolin-6-yl}-
prop-2-ynyl)-
carbamic acid tert-butyl ester
[ 1-(3-Fluoro-benzyl)-1 H-indazol-5-yl]-(6-iodo-quinazolin-4-yl)-amine
hydrochloride
(0.334 g, 0.628 mmol) and Prop-2-ynyl-carbamic acid tent-butyl ester (113 mg,
1.16 equiv.) is
44
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
treated with i-Pr~NH (2 equiv.) in dry THF (4 ml) under N2. Pd(PPh3)a (25 mg,
0.0356 mmol, 5.7
mol%) and solid CuI (5 mol%) are added next, and the mixture is stirred at
r.t. for 3 hours.
Workup: THF is removed under reduced pressure and DCM (10 ml) is added. The
organic layer
is washed with sat. aq. NHaCI solution and brine, dried and concentrated.
Chromatography on
silica (EtOAc/hexanes) affords the desired pure product (293 mg, 89%).
Step E: [6-(3-Amino-prop-1-ynyl)-quinazolin-4-yl]-[1-(3-fluoro-benzyl)-1H-
indazol-5-yl]-amine
(3-{4-[ 1-(3-Fluoro-benzyl)-1 H-indazol-5-ylamino]-quinazolin-6-yl}-prop-2-
ynyl)-
carbamic acid tert-butyl ester (285 mg, 0.545 mmol) is suspended in DCM (6 ml)
and TFA (6
ml) is added dropwise. The reaction is stirred at r.t. fox 2 hours. The
solvents are removed in a N2
stream, DCM (IO ml) is added, and the organic layer is treated with sat. aq.
NaHC03 and brine,
dried, and concentrated to afford the pure product (197.6 mg, 86%).
Step F: 1-(3-{4-[1-(3-Fluoro-benzyl)-1H-indazol-5-ylamino]-quinazolin-6-yl}-
prop-2-ynyl)-2-
phenyl-3-cyano-isourea
[6-(3-Amino-prop-1-ynyl)-quinazolin-4-yl]-[ 1-(3-fluoro-benzyl)-1 H-indazol-5-
yl]-amine
(280 mg, 0.663 mmol) is treated with diphenyl cyanocarbonimidate (163 mg,
1.031 equiv.) in a
mixture of i-PrOH (12 ml) and THF (4 ml). Stir mixture overnight, then
concentrate to dryness
and chromatograph on silica (EtOAc/hexanes) to obtain 186.3 mg (49.6 %) of
pure desired
product.
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Step G: N-cyano-N'-(3-{4-[1-(3-Fluoro-benzyl)-1H-indazol-5-ylamino]-quinazolin-
6-yl}-prop-
.2-ynyl)-morpholine-4-carboxamidine
Material from step F (9 mg, 0.016 mmol) is placed in a reaction vial and
dissolved in 2
ml of a 1:1 THF:i-PrOH mixture. Morpholine (0.08 mmol) is added at r.t, and
the sealed vial is
heated in an oil bath at 80 °C. Reaction progress at 80 °C is
followed by LC/MS, and the reaction
is stopped after reaching 90% conversion (3 hours). Chromatography of the
crude on silica
(MeOH/EtOAc) affords pure desired product (1.8 rng, 20 %). MS ESI (+) m/z 560
(M+1)
detected; 1H NMR (400 MHz, deuterated acetone containing 10% deuterated
methanol) & 8.58
(s, 1 H), 8.55 (s, 1 H), 8.39 (s, 1 H), 8.11 (s, 1 H), 7.84-7.76 (m, 4H), 7.65
(d, 1 H), 7.37 (q; 1 H),
7.12 (d, IH), 7.04 (m, 2H).
Example 10
F
N
~N
~CN
N HN
-MeO~N~N ~ ~ ~ N
H ~ i NJ
N-cyano-N'-(3- { 4-[ 1-(3-Fluoro-benzyl)-1 H-indazol-5-ylamino]-quinazo lin-6-
yl } -allyl)-
N"-(2-methoxy-ethyl)-N"-methylguanidine
46
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Step A: (3-{4-[1-(3-Fluoro-benzyl)-1H-indazol-5-ylamino]-quinazolin-6-yl}-
allyl)-carbamic
acid tent-butyl ester
To a cold (ice-water bath) Red-A1 (0.52 m1, 65% wt solution in toluene, 1.74
mmol)
solution in THF (3 ml) added a solution of the s.m. alkyne (350 mg, 0.670
mmol) in THF (4 ml).
Stir at 0 °C for 2.5 hours. Reaction is quenched with 10% aqueous
potassium carbonate solution
and diluted with distilled water. The mixture is extensively extracted with
EtOAc and DCM,
dried, and concentrated, Yield after chromatography (silica, EtOAclhexanes) is
215.4 mg of pure
product (61 %).
Step B: [6-(3-Amino-propenyl)-quinazolin-4-yl]-[1-(3-fluoro-benzyl)-1H-inda~ol-
5-yl]-amine
The desired product was obtained through a procedure analogous to the one
outlined in
Example 9, step E.
Step C: 1-(3-{4-[I-(3-Fluoro-benzyl)-1H-indazol-5-ylamino]-quinazolin-6-yl}-
allyl)-2-phenyl-3-
cyano-isourea
The desired material was obtained through a procedure analogous to the one
outlined in
Example 9, step F.
47
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Step D: N-cyano-N'-(3- f 4-[I-(3-Fluoro-benzyl)-1H-indazol-5-ylaminoJ-
quinazolin-6-yl}-allyl)-
N"-(2-methoxy-ethyl)-N"-methylguanidine
Material from step C (10 mg, 0.0168 mmol) dissolved in 2ml of a 1:1 THF:i-PrOH
mia~ture and treated with 10 equivalents of MeHNCHaCH2OMe and heat to 80
°C. The reaction
was monitored by LC/MS. Up to five additional equivalents of amine can be
added when
necessary to drive the reaction forward. The reaction was stopped (by cooling
vial to room
'temperature) when the conversion exceeded 60% (80-90% conversion is usually
attained before
end of the reaction). Chromatography of the crude on silica (0-10% MeOH-EtOAc)
followed by
preparative TLC (silica, MeOH/EtOAc) afforded 3.2 mg (34%) of the desired
product. MS ESI
(+) m/z 564 (M+1) detected;'H NMR (400 MHz, denterated acetone) & 9.34 (s,
1H), 8.56 (s,
1 H), 8.45 (d, l H), 8.44 (m, 1 H), 8.10 (s, 1 H), 7.97 (dd, 1 H), 7.77 '(m,
2H), 7.64 (d, 1 H), 7.37 (m,
IH), 7.12~(d, 1H), 7.05 (m, 2H), 6.79 (m, 2H), 6.55 (dt, 1H), 5.73 (s, 2H),
4.31 (m, 2H), 3.36 (s,
3H), 3.12 (s, 3H), 3.63 (m, 4H).
Ezample 11
The extent to which the compounds of the present invention modulate ErbB
kinase
activity can be determined using the following enzyme-linked immunosorbent
assay (ELISA),
which employs a microtiter plate coated with a protein tyrosine kinase
specific polymer
substrate. The phosphorylation reaction is performed on poly-Glu-Tyr 4:1 (PGT)
coated
microtiter plates in the presence on Mgr, ATP and EGFR. The phosphorylated
polymer
substrate is detected with a phosphotyrosine specific monoclonal antibody
conjugated to
48
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
.horseradish peroxidase (HRP). Chromogenic substrate {TMB) color is
quantitiated by
spectrophotometry.
The assay is performed in a 96-well microtiter plate (Immunlon 4, available
from
Dynex). To prepare the plate, 100 pL of 0.25 mglmL Poly (Glu,Tyr) 4:1 Sodium
Salt (available
from Sigma, Catalog Number P027S) in phosphate buffered saline (PBS) is~added
to each well
and the plates are sealed. Following incubation overnight at ambient
temperature, this coating
solution is removed and the plates are washed three times with 300 p,L of
0.1°!o Tween 20
(available from Sigma, Catalog Number P2287) in PBS. If not using immediately,
the coated
microtiter plates may be stored at 2-8 °C with 150 p.L of 0.1°!o
Tween 20 in PBS in each well.
The compound to be tested is dissolved in DMSO at an initial concentration of
1.0 mM.
This initial concentration is diluted 1:25 in DMSO, and the resulting solution
is further serially
diluted 1:5 eight times in DMSO. To 10 ~.L of the initial concentration and
each dilution are
added 240 ~L Reaction Buffer (SO mM HEPES, 125 mM NaCI, 24 mM MgCl2, 0.1 mM
Na3V04
(boiled at pH 10 until colorless -- approximately 10 minutes - and cooled
prior to use), pH 7.3,
filtered through a 0.2 micron filter). 25 ~L of each compound solution
(4°!o DMSO in Reaction
Buffer for control) is placed in a separate microtiter plate well, along with
50 p,L Reaction Buffer
+ ATP ( 15 ~L 10 of mM ATP added to 5 mL Reaction Buffer) and 25 p,L Reaction
Buffer into
which a catalytic amount of baculovirus ErbB2 has been added. The plate is
then coveied and
incubated for 30 minutes at room temperature, after which time all liquid is
aspirated from each
well. The plate is washed three times with 300 p.L of 0.1°!o Tween in
PBS. Residual wash
solution is removed by inverting the plate and blotting on a paper towel.
49
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
To each well is then added 100 ~L of PBS containing 3% bovine serum albumin
(protease-free, IgG-free, Jackson Catalog Number 001-000-162), 0.05% Tween 20
and 0.2
~glmL anti-phosphotyrosine horseradish peroxidase (available from Zymed,
Laboratories, Inc.,
Catalog Number 03-?720). The plate is covered and incubated for 30 minutes at
room
temperature, after which time all liquid is aspirated from each well. The
plate is washed three
times with 300 p.L of 0.1 % Tween in PBS. Residual wash solution is removed by
inverting the
plate and blotting on a paper towel. To each well is then added 100 pL of TMB
peroxidase
substrate system (KPL Catalog Number SO-?6-00), and the plate is allowed to
incubate for 25
minutes at room temperature, at which time the reaction is stopped by the
addition of 100 p,L of
1M phosphoric acid to each well. The plate is tapped gently to ensure mixing.
Within about thirty minutes after the reaction is stopped, the optical density
at 450 nm of
each well is determined using a microtiter plate reader. A dose response curve
is generated by
plotting optical density versus compound concentration. IC$~ is calculated
from this curve using
methods known in the art. '
With this assay, the following ICso values of selected compounds of the
present invention
set forth in Table 1 below were determined.
CA 02506503 2005-05-17
WO 2004/046101 PCT/US2003/035670
Table 1
Example ICso (nm)
#
1 85
' 2 410
3 8
4 13
31
6 14
7 33
8 40
9 12
17
Obviously, numerous modifications and variations of the present invention are
possible in
light of the above teachings. It is therefore to be understood that, within
the scope of the
appended claims, the invention may be practiced otherwise than as specifically
described herein.
S1
