U.S. patent application number 15/292468 was filed with the patent office on 2017-02-02 for macrocyclic compounds as alk, fak and jak2 inhibitors.
The applicant listed for this patent is Cephalon, Inc.. Invention is credited to Henry J. Breslin, Bruce D. Dorsey, Gregory R. Ott.
Application Number | 20170027948 15/292468 |
Document ID | / |
Family ID | 45908071 |
Filed Date | 2017-02-02 |
United States Patent
Application |
20170027948 |
Kind Code |
A1 |
Breslin; Henry J. ; et
al. |
February 2, 2017 |
MACROCYCLIC COMPOUNDS AS ALK, FAK AND JAK2 INHIBITORS
Abstract
The present invention provides compounds of Formula I
##STR00001## or a pharmaceutically acceptable salt forms thereof,
wherein R1, R2, R3, R4, R5, A and X are as defined herein, methods
of treatment and uses thereof.
Inventors: |
Breslin; Henry J.;
(Lansdale, PA) ; Dorsey; Bruce D.; (Ambler,
PA) ; Ott; Gregory R.; (Media, PA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Cephalon, Inc. |
Frazer |
PA |
US |
|
|
Family ID: |
45908071 |
Appl. No.: |
15/292468 |
Filed: |
October 13, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14025986 |
Sep 13, 2013 |
9487529 |
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15292468 |
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PCT/US2012/028856 |
Mar 13, 2012 |
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14025986 |
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61453259 |
Mar 16, 2011 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/5377 20130101;
A61K 31/529 20130101; C07D 487/08 20130101; C07D 487/18
20130101 |
International
Class: |
A61K 31/529 20060101
A61K031/529; A61K 31/5377 20060101 A61K031/5377 |
Claims
1. A method of treating a subject suffering from an ALK-, FAK- or
JAK2-mediated disorder or condition comprising administering to the
subject a therapeutically effective amount of a compound of Formula
I ##STR00035## wherein: A is H, cyano, F, Cl, Br, CH.sub.3 or
CF.sub.3; R1 is H, heterocyclyl, heteroaryl, carbocyclyl, aryl,
C.sub.1-6alkyl, C.sub.1-6alkoxy, carbocyclyloxy, or
C.sub.1-6alkylamino, where, when R1 is a nitrogen containing
heterocyclyl, the nitrogen may be substituted with C.sub.1-6alkyl,
hydroxy(C.sub.2-3)alkyl, dihydroxy (C.sub.3)alkyl,
C.sub.1-6alkoxy(C.sub.2-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.2-3)alkyl,
(C.sub.1-6 alkyl)sulfonyl, amino(C.sub.2-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl, where the
alkyl groups of
N,N-di-(C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl may be the
same or different, heteroaryl or heterocyclyl, and where, when R1
is heterocyclyl, heteroaryl, carbocyclyl, or aryl, such
heterocyclyl, heteroaryl, carbocyclyl, or aryl may be
unsubstituted, or substituted with one or two substituents
independently selected from the group consisting of C.sub.1-6alkyl,
hydroxyl, hydroxy(C.sub.1-3)alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxy(C.sub.1-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
(C.sub.1-6)alkylsulfonyl, fluorine, carboxy,
amino(C.sub.1-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl, where the
alkyl groups of
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl may be the
same or different, and heterocyclyl, and where, when R1 is
C.sub.1-6alkyl, C.sub.1-6alkoxy, or C.sub.1-6alkylamino, the alkyl
groups of such C.sub.1-6alkyl, C.sub.1-6alkoxy, or
C.sub.1-6alkylamino may be unsubstituted, or substituted with one,
two or three substituents independently selected from the group
consisting of heterocyclyl, heteroaryl, hydroxyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
carboxy, aminocarbonyl, N--(C.sub.1-6)alkylaminocarbonyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or different,
aminosulfonyl, and C.sub.1-6alkylsulfonyl; R2 is H, heterocyclyl,
heteroaryl, carbocyclyl, aryl, C.sub.1-6alkyl, C.sub.1-6alkoxy, or
C.sub.1-6alkylamino, where, when R2 is a nitrogen containing
heterocyclyl, the nitrogen may be substituted with H,
C.sub.1-6alkyl, hydroxy(C.sub.2-3)alkyl, dihydroxy (C.sub.3)alkyl,
C.sub.1-6alkoxy(C.sub.2-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.2-3)alkyl,
(C.sub.1-6 alkyl)sulfonyl, amino(C.sub.2-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl, where the
alkyl groups of
N,N-(di-C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl may be the
same or different, heteroaryl or heterocyclyl, and where, when R2
is heterocyclyl, heteroaryl, carbocyclyl, or aryl, such
heterocyclyl, heteroaryl, carbocyclyl, or aryl may be
unsubstituted, or substituted with one or two substituents
independently selected from the group consisting of C.sub.1-6alkyl,
hydroxyl, hydroxy(C.sub.1-3)alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxy(C.sub.1-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
(C.sub.1-6)alkylsulfonyl, fluoro, carboxy,
amino(C.sub.1-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl, where the
alkyl groups of
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl may be the
same or different, and heterocyclyl, and where, when R2 is
C.sub.1-6alkyl, C.sub.1-6alkoxy, or C.sub.1-6alkylamino, the alkyl
groups of such C.sub.1-6alkyl, C.sub.1-6alkoxy, or
C.sub.1-6alkylamino may be unsubstituted, or substituted with one,
two or three substituents independently selected from the group
consisting of heterocyclyl, heteroaryl, hydroxyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
carboxy, aminocarbonyl, N--(C.sub.1-6)alkylaminocarbonyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or different,
aminosulfonyl, and C.sub.1-6alkylsulfonyl; or R1 and R2, together
with the phenyl ring to which they are attached, form a five- to
eight-membered monocarbocyclic or monoheterocyclic ring, where the
ring so formed by R1 and R2, may be unsubstituted, or substituted
with one to three substituents independently selected from the
group consisting of C.sub.1-6alkyl, amino(C.sub.1-3)alkyl,
N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl, where the alkyl
groups of N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl may be the
same or different, alkoxy(C.sub.1-6)alkyl, hydroxyl,
hydroxy(C.sub.1-6)alkyl, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl,
heterocyclyl, amino, (C.sub.1-6)alkylamino,
di-(C.sub.1-6)alkylamino, where the alkyl groups of
di-(C.sub.1-6)alkylamino may be the same or different, carboxy,
aminocarbonyl, N--(C.sub.1-6)alkylaminocarbonyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or different,
aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, and aminosulfonyl; R3 is H, C.sub.1-6alkoxy, or
Cl; where at least one of R1 and R3 is other than H; R4 and R5,
independently, are H, C.sub.1-6alkoxy, aminocarbonyl,
N--(C.sub.1-6alkyl)aminocarbonyl,
N,N-di-(C.sub.1-6alkyl)aminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminocarbonyl may be the same or different,
(C.sub.1-6alkyl)sulfonyl, aminosulfonyl,
N--(C.sub.1-6alkyl)aminosulfonyl,
N,N-di-(C.sub.1-6alkyl)aminosulfonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminosulfonyl may be the same or different,
N--[(C.sub.1-6alkyl)sulfonyl]amino,
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino, where the
alkyl groups of
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino may be the
same or different, (C.sub.1-6alkyl).sub.2phosphinyl, where the
alkyl groups of (C.sub.1-6alkyl).sub.2phosphinyl may be the same or
different, or heteroaryl, where, when R4 or R5 is heteroaryl, such
heteroaryl may be unsubstituted, or substituted with one or two
substituents independently selected from the group consisting of
C.sub.1-6alkyl, hydroxyl, hydroxy(C.sub.1-3)alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxy(C.sub.1-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
(C.sub.1-6alkyl)sulfonyl, fluorine, carboxy, aminocarbonyl,
N--(C.sub.1-6)alkylaminocarbonyl, and
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or different,
and where, when R4 or R5 is N--(C.sub.1-6alkyl)aminocarbonyl or
C.sub.1-6alkoxy, the alkyl groups of such
N--(C.sub.1-6alkyl)aminocarbonyl or C.sub.1-6alkoxy may be
unsubstituted, or substituted by one, two or three substituents
independently selected from the group consisting of heterocyclyl,
hydroxyl, amino, (C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino,
where the alkyl groups of di-(C.sub.1-6)alkylamino may be the same
or different, cyano, carboxy, aminocarbonyl,
N--(C.sub.1-6alkyl)aminocarbonyl,
N,N-di-(C.sub.1-6alkyl)aminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminocarbonyl may be the same or different,
or R4 and R5, together with the phenyl ring to which the are
attached, form a five- to eight-membered monoheterocyclic ring,
where the ring so-formed by R4 and R5 may be unsubstituted, or
substituted with one or two substituents independently selected
from the group consisting of C.sub.1-6alkyl, amino(C.sub.1-3)alkyl,
N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl, where the alkyl
groups of N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl may be the
same or different, alkoxy(C.sub.1-6)alkyl, hydroxyl,
hydroxy(C.sub.1-6)alkyl, (C.sub.1-6alkyl)sulfonyl,
(C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, heterocyclyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
carboxy, aminocarbonyl, N--(C.sub.1-6alkyl)aminocarbonyl,
N,N-di-(C.sub.1-6alkyl)aminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminocarbonyl may be the same or different,
aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, and aminosulfonyl; and --X-- is --CH.dbd.CH--,
--CH.sub.2CH.sub.2--, --NH--CO--, --CONH-- or --(CH(OH)).sub.2--;
or a pharmaceutically acceptable salt thereof.
2. The method of claim 1 wherein the ALK-, FAK- or JAK2-mediated
disorder or condition is cancer.
3. The method of claim 1 wherein the ALK-, FAK- or JAK2-mediated
disorder or condition is selected from colon cancer, breast cancer,
renal cancer, lung cancer, hemangioma, squamous cell myeloid
leukemia, melanoma, glioblastoma, and astrocytoma.
4. A method of treating a proliferative disorder in a subject in
need thereof, comprising administering to the subject a
therapeutically effective amount of a compound of Formula I
##STR00036## wherein: A is H, cyano, F, Cl, Br, CH.sub.3 or
CF.sub.3; R1 is H, heterocyclyl, heteroaryl, carbocyclyl, aryl,
C.sub.1-6alkyl, C.sub.1-6alkoxy, carbocyclyloxy, or
C.sub.1-6alkylamino, where, when R1 is a nitrogen containing
heterocyclyl, the nitrogen may be substituted with C.sub.1-6alkyl,
hydroxy(C.sub.2-3)alkyl, dihydroxy (C.sub.3)alkyl,
C.sub.1-6alkoxy(C.sub.2-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.2-3)alkyl,
(C.sub.1-6 alkyl)sulfonyl, amino(C.sub.2-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl, where the
alkyl groups of
N,N-di-(C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl may be the
same or different, heteroaryl or heterocyclyl, and where, when R1
is heterocyclyl, heteroaryl, carbocyclyl, or aryl, such
heterocyclyl, heteroaryl, carbocyclyl, or aryl may be
unsubstituted, or substituted with one or two substituents
independently selected from the group consisting of C.sub.1-6alkyl,
hydroxyl, hydroxy(C.sub.1-3)alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxy(C.sub.1-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
(C.sub.1-6)alkylsulfonyl, fluorine, carboxy,
amino(C.sub.1-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl, where the
alkyl groups of
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl may be the
same or different, and heterocyclyl, and where, when R1 is
C.sub.1-6alkyl, C.sub.1-6alkoxy, or C.sub.1-6alkylamino, the alkyl
groups of such C.sub.1-6alkyl, C.sub.1-6alkoxy, or
C.sub.1-6alkylamino may be unsubstituted, or substituted with one,
two or three substituents independently selected from the group
consisting of heterocyclyl, heteroaryl, hydroxyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
carboxy, aminocarbonyl, N--(C.sub.1-6)alkylaminocarbonyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or different,
aminosulfonyl, and C.sub.1-6alkylsulfonyl; R2 is H, heterocyclyl,
heteroaryl, carbocyclyl, aryl, C.sub.1-6alkyl, C.sub.1-6alkoxy, or
C.sub.1-6alkylamino, where, when R2 is a nitrogen containing
heterocyclyl, the nitrogen may be substituted with H,
C.sub.1-6alkyl, hydroxy(C.sub.2-3)alkyl, dihydroxy (C.sub.3)alkyl,
C.sub.1-6alkoxy(C.sub.2-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.2-3)alkyl,
(C.sub.1-6 alkyl)sulfonyl, amino(C.sub.2-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl, where the
alkyl groups of
N,N-(di-C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl may be the
same or different, heteroaryl or heterocyclyl, and where, when R2
is heterocyclyl, heteroaryl, carbocyclyl, or aryl, such
heterocyclyl, heteroaryl, carbocyclyl, or aryl may be
unsubstituted, or substituted with one or two substituents
independently selected from the group consisting of C.sub.1-6alkyl,
hydroxyl, hydroxy(C.sub.1-3)alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxy(C.sub.1-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
(C.sub.1-6)alkylsulfonyl, fluoro, carboxy,
amino(C.sub.1-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl, where the
alkyl groups of
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl may be the
same or different, and heterocyclyl, and where, when R2 is
C.sub.1-6alkyl, C.sub.1-6alkoxy, or C.sub.1-6alkylamino, the alkyl
groups of such C.sub.1-6alkyl, C.sub.1-6alkoxy, or
C.sub.1-6alkylamino may be unsubstituted, or substituted with one,
two or three substituents independently selected from the group
consisting of heterocyclyl, heteroaryl, hydroxyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
carboxy, aminocarbonyl, N--(C.sub.1-6)alkylaminocarbonyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or different,
aminosulfonyl, and C.sub.1-6alkylsulfonyl; or R1 and R2, together
with the phenyl ring to which they are attached, form a five- to
eight-membered monocarbocyclic or monoheterocyclic ring, where the
ring so formed by R1 and R2, may be unsubstituted, or substituted
with one to three substituents independently selected from the
group consisting of C.sub.1-6alkyl, amino(C.sub.1-3)alkyl,
N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl, where the alkyl
groups of N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl may be the
same or different, alkoxy(C.sub.1-6)alkyl, hydroxyl,
hydroxy(C.sub.1-6)alkyl, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl,
heterocyclyl, amino, (C.sub.1-6)alkylamino,
di-(C.sub.1-6)alkylamino, where the alkyl groups of
di-(C.sub.1-6)alkylamino may be the same or different, carboxy,
aminocarbonyl, N--(C.sub.1-6)alkylaminocarbonyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or different,
aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, and aminosulfonyl; R3 is H, C.sub.1-6alkoxy, or
Cl; where at least one of R1 and R3 is other than H; R4 and R5,
independently, are H, C.sub.1-6alkoxy, aminocarbonyl,
N--(C.sub.1-6alkyl)aminocarbonyl,
N,N-di-(C.sub.1-6alkyl)aminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminocarbonyl may be the same or different,
(C.sub.1-6alkyl)sulfonyl, aminosulfonyl,
N--(C.sub.1-6alkyl)aminosulfonyl,
N,N-di-(C.sub.1-6alkyl)aminosulfonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminosulfonyl may be the same or different,
N--[(C.sub.1-6alkyl)sulfonyl]amino,
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino, where the
alkyl groups of
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino may be the
same or different, (C.sub.1-6alkyl).sub.2phosphinyl, where the
alkyl groups of (C.sub.1-6alkyl).sub.2phosphinyl may be the same or
different, or heteroaryl, where, when R4 or R5 is heteroaryl, such
heteroaryl may be unsubstituted, or substituted with one or two
substituents independently selected from the group consisting of
C.sub.1-6alkyl, hydroxyl, hydroxy(C.sub.1-3)alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxy(C.sub.1-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
(C.sub.1-6alkyl)sulfonyl, fluorine, carboxy, aminocarbonyl,
N--(C.sub.1-6)alkylaminocarbonyl, and
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or different,
and where, when R4 or R5 is N--(C.sub.1-6alkyl)aminocarbonyl or
C.sub.1-6alkoxy, the alkyl groups of such
N--(C.sub.1-6alkyl)aminocarbonyl or C.sub.1-6alkoxy may be
unsubstituted, or substituted by one, two or three substituents
independently selected from the group consisting of heterocyclyl,
hydroxyl, amino, (C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino,
where the alkyl groups of di-(C.sub.1-6)alkylamino may be the same
or different, cyano, carboxy, aminocarbonyl,
N--(C.sub.1-6alkyl)aminocarbonyl,
N,N-di-(C.sub.1-6alkyl)aminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminocarbonyl may be the same or different,
or R4 and R5, together with the phenyl ring to which the are
attached, form a five- to eight-membered monoheterocyclic ring,
where the ring so-formed by R4 and R5 may be unsubstituted, or
substituted with one or two substituents independently selected
from the group consisting of C.sub.1-6alkyl, amino(C.sub.1-3)alkyl,
N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl, where the alkyl
groups of N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl may be the
same or different, alkoxy(C.sub.1-6)alkyl, hydroxyl,
hydroxy(C.sub.1-6)alkyl, (C.sub.1-6alkyl)sulfonyl,
(C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, heterocyclyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
carboxy, aminocarbonyl, N--(C.sub.1-6alkyl)aminocarbonyl,
N,N-di-(C.sub.1-6alkyl)aminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminocarbonyl may be the same or different,
aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, and aminosulfonyl; and --X-- is --CH.dbd.CH--,
--CH.sub.2CH.sub.2--, --NH--CO--, --CONH-- or --(CH(OH)).sub.2--;
or a pharmaceutically acceptable salt thereof.
5. The method according to claim 4 wherein the proliferative
disorder is cancer.
6. The method according to claim 4 wherein the proliferative
disorder is selected from colon cancer, breast cancer, renal
cancer, lung cancer, hemangioma, squamous cell myeloid leukemia,
melanoma, glioblastoma, and astrocytoma.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of Ser. No.
14/025,986, filed Sep. 13, 2013, which is a continuation
application of International Application No. PCT/US2012/028856,
filed Mar. 13, 2012, which claims priority to U.S. Provisional
Application Ser. No. 61/453,259, filed Mar. 16, 2011. The
disclosures of the aforementioned applications are incorporated
herein by reference in their entireties for all purposes.
BACKGROUND OF THE INVENTION
[0002] Anaplastic lymphoma kinase (ALK) is an orphan receptor
tyrosine kinase (RTK) originally identified as part of the
nucleophosmin (NPM)-ALK fusion gene in anaplastic large cell
lymphoma (ALCL) with a t(2;5) chromosomal translocation (Morris, S.
W. et al., Science, 1994, 263, 1281-1284; Shiota, M. et al.,
Oncogene, 1994, 9, 1567-1574). ALK expression is mainly restricted
to the central and peripheral nervous systems, implicating a
potential role in the physiological development and function of the
nervous system (Iwahara, T. et al., Oncogene, 1997, 14, 439-449;
Morris, S. W., et al., Oncogene, 1997, 14, 2175-2188). ALK knockout
mice possess a full life span and have no overt abnormalities
(Webb, T. R. et al., Expert Rev. Anti-cancer Ther., 2009, 9,
331-356), suggesting ALK inhibition could be well tolerated without
severe adverse effects. While the physiological role of ALK
receptor has not been well defined, involvement of ALK in the
oncogenesis of various human cancers has been well documented and
characterized. Besides NPM-ALK, various other ALK fusion genes were
subsequently detected in ALCL, inflammatory myofibroblastic tumor
(IMT), diffuse large B-cell lymphoma (DLBCL), systemic
histiocytosis, and most notably, in non-small cell lung cancer
(NSCLC), resulting in the generation of oncogenic ALK fusion
proteins with constitutive phosphorylation/activation of ALK, which
plays causative role in tumorgenesis by aberrant phosphorylation of
intracellular downstream substrates (Webb, T. R. et al., Expert
Rev. Anti-cancer Ther., 2009, 9, 331-356; Palmer, R. H. et al.,
Biochem. J., 2009, 420, 345-361; Chiarle, R. et al., Nature Rev.
Cancer, 2008, 8, 11-23; Mano H., Cancer Sci., 2008, 99, 2349-2355).
In NSCLC, at least seven isoforms of an oncogenic fusion gene
comprised of portions of the echinoderm microtubule-associated
protein-like 4 (EML4) gene and ALK gene were identified in about
3-15% patients examined (Soda, M. et al., Nature, 2007, 448,
561-566; Choi Y. L. et al., Cancer Res., 2008, 68, 4971-4976;
Takeuchi, K. et al., Clin. Cancer Res., 2009, 15, 3143-3149).
Experimental data indicate that inhibition of ALK could markedly
impair the growth of ALK-positive lymphoma and lung cancer cells in
vitro and in vivo, indicating that ALK-positive ALCL and NSCLC
cells displayed "ALK oncogene addiction" (Piva, R. et al., Blood,
2006, 107, 689-697; Wan, W. et al., 2006; Galkin, A. V. et al.,
Proc. Natl. Acad. Sci. USA, 2007, 104, 270-275; Christensen, J. G.
et al., Mol. Cancer. Ther., 2007, 6, 3389-3395; Soda, M. et al.,
Proc. Natl. Acad. Sci. USA, 2008, 105, 19893-19897; Koivunen, J. P.
et al., Clin. Cancer Res., 2008, 14, 4275-4283). Recently, it has
also been reported that germline mutations in ALK are the cause of
most hereditary neuroblastoma cases, and ALK activation by mutation
and/or gene amplification is functionally relevant in high-risk
sporadic neuroblastoma (Mosse, Y. P. et al., Nature, 2008, 455,
930-936; Chen, Y. et al., Nature, 2008, 455, 971-974; George, R. E.
et al., Nature, 2008, 455, 975-978; Janoueix-Lerosey, I. et al.,
Nature, 2008, 455, 967-970; McDermott, U. et al., Cancer Res.,
2008, 68, 3389-3395; Passoni, L. et al., Cancer Res., 2009, 69,
7338-7346). Attenuation and inhibition of ALK activating mutants or
wild type (WT) receptor resulted in profound growth inhibition in
human neuroblastoma cell lines (Mosse, Y. P. et al., Nature, 2008,
455, 930-936; Chen, Y. et al., Nature, 2008, 455, 971-974; George,
R. E. et al., Nature, 2008, 455, 975-978; Janoueix-Lerosey, I. et
al., Nature, 2008, 455, 967-970; McDermott, U. et al., Cancer Res.,
2008, 68, 3389-3395; Passoni, L. et al., Cancer Res., 2009, 69,
7338-7346), indicating that the ALK receptor, either the activating
mutants or overexpressed WT form, is a critical player in
neuroblastoma development. Altogether, these findings indicate that
ALK is a major therapeutic target for human cancers and inhibition
of ALK with a small molecule ALK inhibitor would offer a
potentially more effective and less toxic therapy for patients with
ALK-positive tumors than conventional chemotherapy.
[0003] ALK belongs to the insulin receptor (IR) RTK superfamily,
which includes insulin-like growth factor-1 receptor (IGF-1R),
insulin related receptor (IRR) and leukocyte tyrosine kinase (LTK)
(Iwahara, T. et al., Oncogene, 1997, 14, 439-449; Morris, S W. et
al., Oncogene, 1997, 14, 2175-2188). Of these IR family members,
inhibition of IR itself poses a potential liability due to its
involvement in glucose uptake and metabolism (Espinal, Trends
Biochem Sci, 1988, 13, 367-368; Saperstein, R. et al.,
Biochemistry, 1989, 28, 5694-5701). Development of ALK inhibitors
with selectivity over IR is, therefore, desirable.
[0004] Focal adhesion kinase (FAK) is an evolutionarily conserved
non-receptor tyrosine kinase localized at focal adhesions, sites of
cellular contact with the extracellular matrix that functions as a
critical transducer of signaling from integrin receptors and
multiple receptor tyrosine kinases (Parsons, J. T. et al., Clin.
Cancer Res., 2008, 14, 627-632; Han, E. K. and McGonigal, T.,
Anticancer Agents Med. Chem. 2007, 7: 681-684; Schwock, J. et al.,
Expert Opin. Ther. Targets, 2010, 14, 77-94). The
integrin-activated FAK forms a binary complex with Src which can
phosphorylate other substrates and trigger multiple signaling
pathways. Given the central role of FAK in mediating signal
transduction with multiple SH2- and SH3-domain effector proteins
(Mitra, S. K. et al., Mol. Cell Biol., 2005, 6, 56-68), activated
FAK plays a central role in mediating cell adhesion, migration,
morphogenesis, proliferation and survival in normal and malignant
cells (Mitra, S. K. et al., Mol. Cell Biol., 2005, 6, 56-68;
McClean, G. W. et al., Nature Rev. Cancer, 2005, 5, 505-515; Han,
E. K. and McGonigal, T., Anticancer Agents Med. Chem. 2007, 7:
681-684; Chatzizacharias, N. A. et al., Histol. Histopathol., 2008,
23, 629-650). Compared to normal quiescent cells, FAK
over-expression and activation is a hallmark of multiple solid
tumors, particularly those with a propensity for bone metastasis,
specifically breast cancer, ovarian cancer, NSCLC, prostate cancer,
and head/neck squamous cell carcinoma (HNSCC) (Zhao, J. and Guan J.
L., Cancer Metastasis Rev., 2009, 28, 35-49; Schwock, J. et al.,
Expert Opin. Ther. Targets, 2010, 14, 77-94). Moreover, FAK
over-expression and activation mediate anchorage-independent cell
survival and are associated with an enhanced invasive and
metastatic phenotype and tumor angiogenesis in these malignancies
(Owens, L. V. et al., Cancer Res., 1995, 55, 2752-2755., Kornberg,
I. J., et al., Head and Neck, 1998, 20: 634-639; McClean, G. W. et
al., Nature Reviews Cancer, 2005, 5, 505-515; Han, E. K. and
McGonigal, T. Anticancer Agents Med. Chem. 2007, 7: 681-684;
Chatzizacharias, N. A. et al., Histol. Histopathol., 2008, 23,
629-650). Elevated FAK levels in tumors are often caused by
amplification of the FAK gene locus i.e. in breast carcinomas, and
the critical role of FAK in the metastatic progression of breast
cancer has been demonstrated pre-clinically in conditional knockout
studies (van Nimwegen, M. J. et al., Cancer Res., 2005, 65,
4698-4706; Pylayeva, Y. et al., J. Clin. Invest., 2009, 119,
252-266). FAK activation also protects tumor cells from
chemotherapy-induced apoptosis, contributing further to tumor
survival and resistance (Han, E. K. and McGonigal, T. Anticancer
Agents Med. Chem. 2007, 7, 681-684; Haider, J. et al., Clin. Cancer
Res., 2006, 12, 4916-4924). Multiple proof-of-concept studies
conducted in various solid tumors using siRNA (Haider, J. et al.,
Clin. Cancer Res., 2006, 12, 4916-4924), dominant-negative FAK
(Kohno, M. et al, Int. J. Cancer. 2002, 97, 336-343) and small
molecule FAK inhibitors (Haider, J. et al., Cancer Res., 2007, 67,
10976-10983; Roberts, W. G. et al., Cancer Res., 2008, 68,
1935-1944; Bagi, C. M. et al., Cancer, 2008, 112, 2313-2321) have
provided pre-clinical support for the therapeutic utility of FAK
inhibition as an anti-tumor/anti-angiogenic strategy, particularly
for androgen-independent prostate cancers, breast cancers, and
HNSCC.
[0005] The Janus kinase (JAK)/Signal transducers and activators of
transcription (STAT) pathway is the major signaling cascade
downstream from cytokine receptors and growth factor receptors
including growth hormone, prolactin and leptin (Rane, S. G. et al.,
Oncogene 2002, 21, 3334-3358; Schindler, C. et al., J. Biol. Chem.
2007, 282, 20059-20066; Baker, S. J. et al., Oncogene, 2007, 15,
6724-6737). The signaling cascade consists of the family of
non-receptor tyrosine kinases, JAK and transcription factors, STAT.
Activated JAK phosphorylate and activate STAT, allowing formation
of homo- and heterodimers that translocate to the nucleus to
regulate the transcription of STAT-dependent genes. In addition,
STAT can be directly phosphorylated by non-receptor tyrosine
kinases like Src or Abl. Under normal physiological conditions
ligand-dependent activation of JAK/STAT signaling is transient and
tightly regulated (Alexander, W. S., Nature Rev. Immunol., 2002, 2,
1-7; Shuai, K. et al., Nature Rev. Immunol., 2003, 3: 900-910).
Constitutive activation of JAK and STAT was detected in a wide
spectrum of human cancers, both solid and hematopoietic, and often
correlated with a more malignant and metastatic phenotype and
refractory tumors (Ferrajoli, A. et al., Curr. Cancer Drug Targets,
2006, 6, 671-9; Yu, H. et al., Nature Rev. Cancer, 2004, 4,
97-105). In most tumors JAK2/STAT activation was mediated by a
constitutive expression of cytokines (IL-6, IL-4, GM-CSF) and/or by
inactivation of endogenous repressors of the JAK/STAT pathway,
including members of the suppressor of cytokine signaling (SOCS)
family or phosphatase SHP-1 due to promoter methylation or specific
deletions (Yoshikava, H. et al., Nature Genetics, 2001, 28, 29-35;
Weber, A., at al., Oncogene, 2005, 24, 6699-708; Melzner, I., et
al., Oncogene, 2005, 24, 6699-708; Weniger, M. et al., Oncogene,
2006, 25, 2679-84). In some tumors, activating mutations in JAK1
(Flex, E. et al., J. Exp. Med. 2008, 205, 751-758; Hornakova, T. et
al., J. Biol. Chem. 2009, 384, 6773-6781), JAK2, JAK3 or JAK2
chimeric molecules were directly implicated in tumorigenesis. In
addition, amplification of the JAK2 locus was found in 35% of
Hodgkin lymphoma (HL) and 50% of primary mediastinal B-cell
lymphoma (PMBL) cases (Melzner, I, et al., Blood, 2005, 105,
2535-2542). The ectopic expression of JAK1, JAK2 and JAK3, as well
as STAT3 and STAT5 resulted in oncogenic transformation in
recipient cells, demonstrating that the activated JAK2/STAT pathway
was sufficient to mediate oncogenesis in various solid and
hematological tumors (Bromberg, J. et al., Cell, 1999, 98, 295-303;
Knoops, L. et al., Oncogene, 2008, 27, 1511-9; Scheeren, F. A. et
al., Blood. 2008, 111, 4706-4715). In multiple studies, inhibition
of JAK2/STAT signaling in various tumor cells, including prostate,
breast, colon and lung carcinomas, gliomas as well as leukemias and
lymphomas resulted in inhibition of growth, induction of apoptosis
and suppression of tumor growth in vivo (Yu, H. et al., Nature Rev.
Cancer, 2004, 4, 97-105; Li, H. et al., Cancer Res. 2004, 64,
4774-4782; Iwamaru, A. et al., Oncogene, 2007, 26, 2435-2444; Gao,
S., et al., J. Clin. Invest., 2007, 117, 3846-3856; Ding, B. et
al., Blood, 2008, 111, 1515-1523,). Pre-clinical studies have
demonstrated that constitutively activated JAK2/STAT signaling in
tumor cells not only promoted uncontrolled cell proliferation and
anti-apoptotic signaling, but also mediated tumor immune evasion
and angiogenesis (Kortylewski, R., et al., Nat. Med., 2005, 11,
1314-21; Nefedova, Y. et al., Curr. Cancer Drug Targets, 2007, 7,
71-77). Therefore, inhibitors of JAK/STAT signaling would
potentially suppress multiple mechanisms underlying tumor formation
and progression.
[0006] Human cancers are notoriously heterogeneous, even in those
well established "oncogene addicted" tumors, since some cancer
cells likely contain additional oncogenic event(s) or redundant
active signaling pathways which may render the cancer cells less
dependent on the primary oncogene for growth and survival (Hanahan,
D. and Weinberg, R. A., Cell, 2000, 100, 57-70). Concomitant
inhibition of the secondary oncogenic event(s) in those cancer
cells would likely lead to increase the efficacy of treatment with
a kinase inhibitor, either by combination therapy or developing a
small molecule inhibitor against both the primary and secondary
targets. For example, FAK was found to be hyperphosphorylated and
activated in a majority of human NSCLC cell lines and contributed
to promote cancer cell invasion and metastasis (Rikova, K., Cell,
2007, 131, 1190-1203). An ALK inhibitor with concomitant FAK
activity may provide additive or synergistic anti-tumor activity
against ALK-positive NSCLC, and potentially additional solid and
hematological tumors containing functional ALK and FAK. Similarly,
JAK2/STAT5 mediated signaling pathway plays an important role in
androgen-independent prostate tumorigenesis (Li, H., et al. Cancer
Res., 2004, 64, 4774-4782; Dagvadorj, A., et al., Endocrinology,
2007, 148, 3089-3101) and FAK activity was found to be critical for
maintaining the invasive and metastatic phenotype of
androgen-independent prostate cancer (Owen, L. V. et al, Cancer
Res., 1995, 55, 2752-2755; Tremblay, L., et al., Int. J. Cancer,
1996, 68, 164-171). Therefore, there is a compelling rationale for
the utility of a dual JAK2/FAK inhibitor to treat this type of
cancer.
[0007] On the other hand, although kinase inhibitors have been
extremely effective in specific patient populations with tumors
containing mutated, oncogenic forms of protein tyrosine kinases
(PTK), clinical studies thus far have shown that some patients
eventually develop resistance to these drugs, either due to the
selection of cancer cells with mutations in the targeted PTK or the
induction of compensatory oncogenic signaling pathways (Shah, N. P.
and Sawyers, C. L., Oncogene, 2003, 22, 7389-7395; Engelman, J. A.
and Settleman, J., Curr. Opin. Genet. Develop., 2008, 18, 1-7; Liu,
J. et al., Leukemia, 2008, 22, 791-799; Desai, J. et al., Clin.
Cancer Res., 2007, 13, 5398-5405; Engelman, J. A. and Janne, P. A.,
Clin. Cancer Res., 2008, 14, 2895-2899). In that regard, a kinase
inhibitor simultaneously inhibiting two or more critical,
non-redundant signaling pathways may prevent or decrease the
incident of resistant tumors to develop.
[0008] A need exists for ALK, FAK and JAK2 inhibitors for use as
pharmaceutical agents.
SUMMARY OF THE INVENTION
[0009] The present invention provides a compound of Formula I
##STR00002##
or a pharmaceutically acceptable salt form thereof, wherein A, R1,
R2, R3, R4, R5 and X are as defined herein.
[0010] The compound of Formula I has ALK, FAK and/or JAK2
inhibitory activity, and may be used to treat ALK-, FAK or
JAK2-mediated disorders or conditions.
[0011] The present invention further provides a pharmaceutical
composition comprising at least one compound of the present
invention together with at least one pharmaceutically acceptable
carrier, diluent, or excipient therefor.
[0012] In another aspect, the present invention provides a method
of treating a subject suffering from an ALK-, FAK or JAK2-mediated
disorder or condition comprising: administering to the subject the
pharmaceutical composition of the present invention.
[0013] The present invention further provides a method of treating
a proliferative disorder in a subject, comprising administering to
the subject a therapeutically effective amount of a compound of the
present invention.
DETAILED DESCRIPTION OF THE INVENTION
I. Definitions
[0014] As used herein, the following terms have the meanings
ascribed to them unless specified otherwise.
[0015] "Alkylamino" or an "alkylamino group" refers to an NH-alkyl
group.
[0016] "Alkoxy" or "alkoxy group" refers to an O-alkyl group.
[0017] "Alkyl" or "alkyl group" refers to a branched or unbranched
saturated hydrocarbon chain. Examples include, but are not limited
to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl,
n-octyl, n-nonyl, n-decyl, isopropyl, tert-butyl, isobutyl, etc.
Alkyl groups typically contain 1-10 carbon atoms, such as 1-6
carbon atoms.
[0018] The term "C.sub.x-y" indicates the number of carbon atoms in
a group. For example, a "C.sub.1-6-alkyl" is an alkyl group having
from one (1) to six (6) carbon atoms.
[0019] The term "cyano" refers to a CN group.
[0020] "Carbocyclyl" or "carbocyclyl group" refers to a
non-aromatic carbocyclic ring system, which may be saturated or
unsaturated, substituted or unsubstituted, and may be monocyclic,
bicyclic, or tricyclic, and may be bridged, spiro, and/or fused.
Examples include, but are not limited to, cyclopropyl,
cyclopropenyl, cyclobutyl, cyclobutenyl, cyclopentyl,
cyclopentenyl, cyclohexyl, cyclohexenyl, norbornyl, norbornenyl,
bicyclo[2.2.1]hexane, bicyclo[2.2.1]heptane, bicyclo[2.2.1]heptene,
bicyclo[3.1.1]heptane, bicyclo[3.2.1]octane, bicyclo[2.2.2]octane,
bicyclo[3.2.2]nonane, bicyclo[3.3.1]nonane, and
bicyclo[3.3.2]decane. Preferably, the cycloalkyl group contains
from 3 to 10 ring atoms. More preferably, the cycloalkyl group
contains from 3 to 7 ring atoms, such as 3 ring atoms, 5 ring
atoms, 6 ring atoms, or 7 ring atoms.
[0021] "Carbocyclyloxy" or a "carbocyclyloxy" group refers to a
carbocyclyl-O-- group.
[0022] "Aryl" or "aryl group" refers to phenyl and 7-15 membered
monoradical bicyclic or tricyclic hydrocarbon ring systems,
including bridged, spiro, and/or fused ring systems, in which at
least one of the rings is aromatic. Aryl groups can be substituted
or unsubstituted. Examples include, but are not limited to, phenyl,
naphthyl, indanyl, 1,2,3,4-tetrahydronaphthalenyl,
6,7,8,9-tetrahydro-5H-benzocycloheptenyl, and
6,7,8,9-tetrahydro-5H-benzocycloheptenyl. Preferably, the aryl
group contains 6 (i.e., phenyl) or 9 to 15 ring atoms. More
preferably, the aryl group contains 6 (i.e., phenyl), 9 or 10 ring
atoms.
[0023] "Heterocyclyl" or "heterocyclyl group" refers to 3-15
membered monocyclic, bicyclic, and tricyclic non-aromatic rings,
which may be saturated or unsaturated, can be substituted or
unsubstituted, may be bridged, spiro, and/or fused, and which
contain, in addition to carbon atom(s), at least one heteroatom,
such as nitrogen, oxygen or sulfur. Examples include, but are not
limited to, tetrahydrofuranyl, pyrrolidinyl, pyrrolinyl,
imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl,
piperidyl, piperazinyl, indolinyl, isoindolinyl, morpholinyl,
thiomorpholinyl, homomorpholinyl, homopiperidyl, homopiperazinyl,
thiomorpholinyl, tetrahydropyranyl, piperidinyl, tetrahydrothienyl,
homopiperidinyl, oxazolidinonyl, dihydropyrazolyl, dihydropyrrolyl,
dihydropyrazinyl, dihydropyridinyl, dihydropyrimidinyl,
dihydrofuryl, dihydropyranyl, quinuclidinyl,
2-oxa-5-azabicyclo[2.2.1]heptane, 8-oxa-3-aza-bicyclo[3.2.1]octane,
3,8-diaza-bicyclo[3.2.1]octane, 2,5-diaza-bicyclo[2.2.1]heptane,
3,8-diaza-bicyclo[3.2.1]octane, 3,9-diaza-bicyclo[4.2.1]nonane and
2,6-diaza-bicyclo[3.2.2]nonane. Preferably, the heterocyclyl group
contains from 3 to 10 ring atoms. More preferably, the heterocycyl
group contains from 3 to 7 ring atoms. More preferably, the
heterocyclyl group contains from 5 to 7 ring atoms, such as 5 ring
atoms, 6 ring atoms, or 7 ring atoms. Unless otherwise indicated,
the foregoing heterocyclyl groups can be C-attached or N-attached
where such is possible and results in the creation of a stable
structure. For example, piperidinyl can be piperidin-1-yl
(N-attached) or piperidin-4-yl (C-attached). A heterocyclyl group
can also include ring systems substituted on ring carbons with one
or more --OH functional groups (which may further tautomerize to
give a ring C.dbd.O group) and/or substituted on a ring sulfur atom
by one (1) or two (2) oxygen atoms to give S.dbd.O or SO.sub.2
groups, respectively.
[0024] "Heteroaryl" or "heteroaryl group" refers to (a) 5 and 6
membered monocyclic aromatic rings, which contain, in addition to
carbon atom(s), at least one heteroatom, such as nitrogen, oxygen
or sulfur, and (b) 7-15 membered bicyclic and tricyclic rings,
which contain, in addition to carbon atom(s), at least one
heteroatom, such as nitrogen, oxygen or sulfur, and in which at
least one of the rings is aromatic. Heteroaryl groups can be
substituted or unsubstituted, and may be bridged, spiro, and/or
fused. Examples include, but are not limited to,
2,3-dihydrobenzofuranyl, 1,2-dihydroquinolinyl,
3,4-dihydroisoquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl,
1,2,3,4-tetrahydroquinolinyl, benzoxazinyl, benzthiazinyl,
chromanyl, furanyl, 2-furanyl, 3-furanyl, imidazolyl, isoxazolyl,
isothiazolyl, oxadiazolyl, oxazolyl, pyridinyl, 2-, 3-, or
4-pyridinyl, pyrimidinyl, 2-, 4-, or 5-pyrimidinyl, pyrazolyl,
pyrrolyl, 2- or 3-pyrrolyl, pyrazinyl, pyridazinyl, 3- or
4-pyridazinyl, 2-pyrazinyl, thienyl, 2-thienyl, 3-thienyl,
tetrazolyl, thiazolyl, thiadiazolyl, triazinyl, triazolyl,
pyridin-2-yl, pyridin-4-yl, pyrimidin-2-yl, pyridazin-4-yl,
pyrazin-2-yl, naphthyridinyl, pteridinyl, phthalazinyl, purinyl,
alloxazinyl, benzimidazolyl, benzofuranyl, benzofurazanyl,
2H-1-benzopyranyl, benzothiadiazine, benzothiazinyl,
benzothiazolyl, benzothiophenyl, benzoxazolyl, cinnolinyl,
furopyridinyl, indolinyl, indolizinyl, indolyl, or 2-, 3-, 4-, 5-,
6-, or 7-indolyl, 3H-indolyl, quinazolinyl, quinoxalinyl,
isoindolyl, isoquinolinyl,
10-aza-tricyclo[6.3.1.0*2,7*]dodeca-2(7),3,5-trienyl,
12-oxa-10-aza-tricyclo[6.3.1.0*2,7*]dodeca-2(7),3,5-trienyl,
12-aza-tricyclo[7.2.1.0*2,7*]dodeca-2(7),3,5-trienyl,
10-aza-tricyclo[6.3.2.0*2,7*]trideca-2(7),3,5-trienyl,
2,3,4,5-tetrahydro-1H-benzo[d]azepinyl,
1,3,4,5-tetrahydro-benzo[d]azepin-2-onyl,
1,3,4,5-tetrahydro-benzo[b]azepin-2-onyl,
2,3,4,5-tetrahydro-benzo[c]azepin-1-onyl,
1,2,3,4-tetrahydro-benzo[e][1,4]diazepin-5-onyl,
2,3,4,5-tetrahydro-1H-benzo[e][1,4]diazepinyl,
5,6,8,9-tetrahydro-7-oxa-benzocycloheptenyl,
2,3,4,5-tetrahydro-1H-benzo[b]azepinyl,
1,2,4,5-tetrahydro-benzo[e][1,3]diazepin-3-onyl,
3,4-dihydro-2H-benzo[b][1,4]dioxepinyl,
3,4-dihydro-2H-benzo[f][1,4]oxazepin-5-onyl,
6,7,8,9-tetrahydro-5-thia-8-aza-benzocycloheptenyl,
5,5-dioxo-6,7,8,9-tetrahydro-5-thia-8-aza-benzocycloheptenyl, and
2,3,4,5-tetrahydro-benzo[f][1,4]oxazepinyl. Preferably, the
heteroaryl group contains 5, 6, or 8-15 ring atoms. More
preferably, the heteroaryl group contains 5 to 10 ring atoms, such
as 5, 6, 9, or 10 ring atoms. A heteroaryl group can also include
ring systems substituted on ring carbons with one or more --OH or
C.dbd.O functional groups and/or substituted on a ring sulfur atom
by one (1) or two (2) oxygen atoms to give S.dbd.O or SO.sub.2
groups, respectively.
[0025] "Hydroxyalkyl" or "hydroxyalky group" refers to an alkyl
group containing an --OH group substituent.
[0026] "Dihydroxyalkyl" or "dihydroxyalkyl group" refers to an
alkyl group containing two --OH group substituents.
[0027] "Alkoxyalkyl" or "alkoxyalkyl group" refers to an alkyl
group containing an alkoxy group substituent.
[0028] "Aminocarbonylalkyl" or "aminocarbonylalkyl group" refers to
an H.sub.2N--C(O)-- alkyl- group.
[0029] "N-alkylaminocarbonylalkyl" or "N-alkylaminocarbonylalkyl
group" refers to an alkyl-NH--C(O)-alkyl- group.
[0030] "N,N-dialkylaminocarbonylalkyl" or
"N,N-dialkylaminocarbonylalkyl group" refers to an
(alkyl)(alkyl)N--C(O)-alkyl- group. In such a group the alkyl
groups substituting the nitrogen may be the same or different.
[0031] "Alkylsulfonylalkyl" or "alkylsulfonylalkyl" group refers to
an alkyl-S(O).sub.2-alkyl-group.
[0032] "Alkylsulfonyl" or "alkylsulfonyl" group refers to an
alkyl-S(O).sub.2-- group.
[0033] "Aminoalkylcarbonyl" or "aminoalkylcarbonyl group" refers to
an H.sub.2N-alkyl-C(O)-- group.
[0034] "N-alkylaminoalkylcarbonyl" or "N-alkylaminoalkylcarbonyl
group" refers to an alkyl-NH-alkyl-C(O)-- group.
[0035] "N,N-dialkylaminoalkylcarbonyl" or
"N,N-dialkylaminoalkylcarbonyl group" refers to an
(alkyl)(alkyl)N-alkyl-C(O)-- group. In such a group the alkyl
groups substituting the nitrogen may be the same or different.
[0036] "Hydroxyl", "hydroxy", "hydroxyl group" or "hydroxyl group"
refers to an --OH group.
[0037] "Alkoxyalky" or "alkoxyalkyl group" refers to an alkyl group
substituted by an alkoxy group.
[0038] "Amino" or "amino group" refers to an --NH.sub.2 group.
[0039] "Alkylamino" or "alkylamino group" refers to an alkyl-N(H)--
group.
[0040] "Dialkylamino" or "dialkylamino" group refers to an
(alkyl)(alkyl)N-- group. In such a group the alkyl groups
substituting the nitrogen may be the same or different.
[0041] "Carboxy", "carboxyl", "carboxy group" or "carboxyl group"
refers to a --COOH group.
[0042] "Aminocarbonyl" or "aminocarbonyl group" refers to an
H.sub.2N--C(O)-- group.
[0043] "N-alkylaminocarbonyl" or "N-alkylaminocarbonyl group"
refers to an alkyl-NH--C(O)-- group.
[0044] "N,N-dialkylaminocarbonyl" or "N,N-dialkylaminocarbonyl"
refers to an (alkyl)(alkyl)N--C(O)-- group. In such a group, the
alkyl group substituents on the nitrogen may be the same or
different.
[0045] "Aminosulfonyl" or "aminosulfonyl group" refers to an
H.sub.2N--S(O).sub.2-- group.
[0046] "N-alkylaminosulfonyl" or "N-alkylaminosulfonyl group"
refers to an (alkyl)N(H)--S(O).sub.2-- group.
[0047] "N,N-dialkylaminosulfonyl" or "N,N-dialkylaminosulfonyl
group" refers to an (alkyl)(alkyl)N--S(O).sub.2-- group. In such a
group the alkyl group substituents on the nitrogen may be the same
of different.
[0048] "N-[(alkyl)sulfonyl]amino" or "N-[(alkyl)sulfonyl]amino
group" refers to an (alkyl)S(O).sub.2--NH-- group.
[0049] "N-[(alkyl)sulfonyl]-N-alkylamino" or
"N-[(alkyl)sulfonyl]-N-alkylamino group" refers to an
(alkyl)S(O).sub.2--N(alkyl)- group. In such a group the alkyl group
substituent on sulfur, and the alkyl group substituent on nitrogen
may be the same or different.
[0050] "Aminoalkyl" or "aminoalkyl group" refers to an
H.sub.2N-alkyl- group.
[0051] "N-alkylaminoalkyl" or "N-alkylaminoalkyl group" refers to
an (alkyl)N(H)-alkyl-group.
[0052] "N,N-dialkyl-aminoalkyl" or "N,N-dialkyl-aminoalkyl group"
refers to an (alkyl)(alkyl)N-alkyl- group. In such a group the
alkyl groups substituting the nitrogen may be the same of
different.
[0053] "(Alkyl).sub.2phosphinyl" or "(alkyl).sub.2phosphinyl group"
refers to a (alkyl)(alkyl)P(O)-- group. In such a group, the alkyl
substituents on phosphorus may be the same or different.
[0054] "Chemically stable" or "stable" refers to a compound that is
sufficiently robust to be isolated to a useful degree of purity
from a reaction mixture. The present invention is directed only to
chemically stable compounds.
[0055] "Pharmaceutical composition" refers to a composition
suitable for administration in medical or veterinary use.
[0056] When lists of alternative substituents include members
which, owing to valency requirements, chemical stability, or other
reasons, cannot be used to substitute a particular group, the list
is intended to be read in context to include those members of the
list that are suitable for substituting the particular group.
[0057] "Functionalized derivative" refers to a compound that
contains at least one additional functional group as compared to a
reference compound. An example of a functionalized derivative of
benzene is bromobenzene. An example of a functionalized derivative
of bromobenzene is 2-bromophenol. Functional groups include, but
are not limited to, halogen, nitro, hydroxy, alkoxy, aryloxy,
ketone, ester, amide, amino, alkylamino, alkyl, double bond, triple
bond, alkoxyalkyl, aminoalkyl, haloalkyl, alkenyl, alkynyl, aryl,
heteroaryl, cycloalkyl, heterocycloalkyl, pseudohalogen, alkylthio,
sulfonyl, alkylsulfonyl, alkylaminosulfonyl, alkylcarbonyloxy,
aminocarbonyloxy, alkylaminocarbonyloxy, alkylcarbonylamino,
alkoxycarbonylamino, alkylsulfonylamino, aminocarbonylamino, and
alkylaminocarbonylamino functional group, and derivatives of these
and other functional groups in which a heteroatom is derivatized
with a removable protecting group.
[0058] "Pharmaceutically acceptable" refers to physiologically
tolerable materials, which do not typically produce an allergic or
other untoward reaction, such as gastric upset, dizziness and the
like, when administered to a mammal.
[0059] "Therapeutically effective amount" refers to an amount of a
compound, or a pharmaceutically acceptable salt thereof, sufficient
to inhibit, halt, or cause an improvement in a disorder or
condition being treated in a particular subject or subject
population. For example in a human or other mammal, a
therapeutically effective amount can be determined experimentally
in a laboratory or clinical setting, or may be the amount required
by the guidelines of the United States Food and Drug
Administration, or equivalent foreign agency, for the particular
disease and subject being treated.
[0060] It should be appreciated that determination of proper dosage
forms, dosage amounts, and routes of administration is within the
level of ordinary skill in the pharmaceutical and medical arts, and
is described below.
[0061] "Subject" refers to a member of the class Mammalia. Examples
of mammals include, without limitation, humans, primates,
chimpanzees, rodents, mice, rats, rabbits, horses, livestock, dogs,
cats, sheep, and cows.
[0062] "Treatment" refers to the acute or prophylactic diminishment
or alleviation of at least one symptom or characteristic associated
or caused by a disorder being treated. For example, treatment can
include diminishment of several symptoms of a disorder or complete
eradication of a disorder.
[0063] "Administering" refers to the method of contacting a
compound with a subject. Modes of "administering" include, but are
not limited to, methods that involve contacting the compound
intravenously, intraperitoneally, intranasally, transdermally,
topically, via implantation, subcutaneously, parentally,
intramuscularly, orally, systemically, and via adsorption.
II. Compounds
[0064] The present invention provides compounds of Formula I
##STR00003##
[0065] wherein:
[0066] A is H, cyano, F, Cl, Br, CH.sub.3 or CF.sub.3;
[0067] R1 is H, heterocyclyl, heteroaryl, carbocyclyl, aryl,
C.sub.1-6alkyl, C.sub.1-6alkoxy, carbocyclyloxy, or
C.sub.1-6alkylamino,
[0068] where, when R1 is a nitrogen containing heterocyclyl, the
nitrogen may be substituted with C.sub.1-6alkyl,
hydroxy(C.sub.2-3)alkyl, dihydroxy (C.sub.3)alkyl,
C.sub.1-6alkoxy(C.sub.2-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.2-3)alkyl,
(C.sub.1-6 alkyl)sulfonyl, amino(C.sub.2-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl, where the
alkyl groups of
N,N-di-(C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl may be the
same or different, heteroaryl or heterocyclyl,
[0069] and where, when R1 is heterocyclyl, heteroaryl, carbocyclyl,
or aryl, such heterocyclyl, heteroaryl, carbocyclyl, or aryl may be
unsubstituted, or substituted with one or two substituents
independently selected from the group consisting of C.sub.1-6alkyl,
hydroxyl, hydroxy(C.sub.1-3)alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxy(C.sub.1-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
(C.sub.1-6)alkylsulfonyl, fluorine, carboxy,
amino(C.sub.1-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl, where the
alkyl groups of
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl may be the
same or different, and heterocyclyl,
[0070] and where, when R1 is C.sub.1-6alkyl, C.sub.1-6alkoxy, or
C.sub.1-6alkylamino, the alkyl groups of such C.sub.1-6alkyl,
C.sub.1-6alkoxy, or C.sub.1-6alkylamino may be unsubstituted, or
substituted with one, two or three substituents independently
selected from the group consisting of heterocyclyl, heteroaryl,
hydroxyl, amino, (C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino,
where the alkyl groups of di-(C.sub.1-6)alkylamino may be the same
or different, carboxy, aminocarbonyl,
N--(C.sub.1-6)alkylaminocarbonyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or different,
aminosulfonyl, and C.sub.1-6alkylsulfonyl;
[0071] R2 is H, heterocyclyl, heteroaryl, carbocyclyl, aryl,
C.sub.1-6alkyl, C.sub.1-6alkoxy, or C.sub.1-6alkylamino,
[0072] where, when R2 is a nitrogen containing heterocyclyl, the
nitrogen may be substituted with H, C.sub.1-6alkyl,
hydroxy(C.sub.2-3)alkyl, dihydroxy (C.sub.3)alkyl,
C.sub.1-6alkoxy(C.sub.2-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.2-3)alkyl,
(C.sub.1-6 alkyl)sulfonyl, amino(C.sub.2-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl, where the
alkyl groups of
N,N-(di-C.sub.1-6alkyl)amino(C.sub.2-3)alkylcarbonyl may be the
same or different, heteroaryl or heterocyclyl,
[0073] and where, when R2 is heterocyclyl, heteroaryl, carbocyclyl,
or aryl, such heterocyclyl, heteroaryl, carbocyclyl, or aryl may be
unsubstituted, or substituted with one or two substituents
independently selected from the group consisting of C.sub.1-6alkyl,
hydroxyl, hydroxy(C.sub.1-3)alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxy(C.sub.1-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
(C.sub.1-6)alkylsulfonyl, fluoro, carboxy,
amino(C.sub.1-3)alkylcarbonyl,
N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl,
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl, where the
alkyl groups of
N,N-(di-C.sub.1-6alkyl)amino(C.sub.1-3)alkylcarbonyl may be the
same or different, and heterocyclyl,
[0074] and where, when R2 is C.sub.1-6alkyl, C.sub.1-6alkoxy, or
C.sub.1-6alkylamino, the alkyl groups of such C.sub.1-6alkyl,
C.sub.1-6alkoxy, or C.sub.1-6alkylamino may be unsubstituted, or
substituted with one, two or three substituents independently
selected from the group consisting of heterocyclyl, heteroaryl,
hydroxyl, amino, (C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino,
where the alkyl groups of di-(C.sub.1-6)alkylamino may be the same
or different, carboxy, aminocarbonyl,
N--(C.sub.1-6)alkylaminocarbonyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or different,
aminosulfonyl, and C.sub.1-6alkylsulfonyl; or
[0075] R1 and R2, together with the phenyl ring to which they are
attached, form a five- to eight-membered monocarbocyclic or
monoheterocyclic ring, where the ring so formed by R1 and R2, may
be unsubstituted, or substituted with one to three substituents
independently selected from the group consisting of C.sub.1-6alkyl,
amino(C.sub.1-3)alkyl, N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl, where the alkyl
groups of N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl may be the
same or different, alkoxy(C.sub.1-6)alkyl, hydroxyl,
hydroxy(C.sub.1-6)alkyl, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl,
heterocyclyl, amino, (C.sub.1-6)alkylamino,
di-(C.sub.1-6)alkylamino, where the alkyl groups of
di-(C.sub.1-6)alkylamino may be the same or different, carboxy,
aminocarbonyl, N--(C.sub.1-6)alkylaminocarbonyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or different,
aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, and aminosulfonyl;
[0076] R3 is H, C.sub.1-6alkoxy, or Cl;
[0077] where at least one of R1 and R3 is other than H;
[0078] R4 and R5, independently, are H, C.sub.1-6alkoxy,
aminocarbonyl, N--(C.sub.1-6alkyl)aminocarbonyl,
N,N-di-(C.sub.1-6alkyl)aminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminocarbonyl may be the same or different,
(C.sub.1-6alkyl)sulfonyl, aminosulfonyl,
N--(C.sub.1-6alkyl)aminosulfonyl,
N,N-di-(C.sub.1-6alkyl)aminosulfonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminosulfonyl may be the same or different,
N--[(C.sub.1-6alkyl)sulfonyl]amino,
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino, where the
alkyl groups of
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino may be the
same or different, (C.sub.1-6alkyl).sub.2phosphinyl, where the
alkyl groups of (C.sub.1-6alkyl).sub.2phosphinyl may be the same or
different, or heteroaryl, where, when R4 or R5 is heteroaryl, such
heteroaryl may be unsubstituted, or substituted with one or two
substituents independently selected from the group consisting of
C.sub.1-6alkyl, hydroxyl, hydroxy(C.sub.1-3)alkyl, C.sub.1-6alkoxy,
C.sub.1-6alkoxy(C.sub.1-3)alkyl, aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, (C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
(C.sub.1-6alkyl)sulfonyl, fluorine, carboxy, aminocarbonyl,
N--(C.sub.1-6)alkylaminocarbonyl, and
N,N-di-(C.sub.1-6)alkylaminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl may be the same or
different,
[0079] and where, when R4 or R5 is N--(C.sub.1-6alkyl)aminocarbonyl
or C.sub.1-6alkoxy, the alkyl groups of such
N--(C.sub.1-6alkyl)aminocarbonyl or C.sub.1-6alkoxy may be
unsubstituted, or substituted by one, two or three substituents
independently selected from the group consisting of heterocyclyl,
hydroxyl, amino, (C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino,
where the alkyl groups of di-(C.sub.1-6)alkylamino may be the same
or different, cyano, carboxy, aminocarbonyl,
N--(C.sub.1-6alkyl)aminocarbonyl,
N,N-di-(C.sub.1-6alkyl)aminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminocarbonyl may be the same or different,
or
[0080] R4 and R5, together with the phenyl ring to which the are
attached, form a five- to eight-membered monoheterocyclic ring,
where the ring so-formed by R4 and R5 may be unsubstituted, or
substituted with one or two substituents independently selected
from the group consisting of C.sub.1-6alkyl, amino(C.sub.1-3)alkyl,
N--(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl, where the alkyl
groups of N,N-di-(C.sub.1-6alkyl)amino(C.sub.1-3)alkyl may be the
same or different, alkoxy(C.sub.1-6)alkyl, hydroxyl,
hydroxy(C.sub.1-6)alkyl, (C.sub.1-6alkyl)sulfonyl,
(C.sub.1-6alkyl)sulfonyl(C.sub.1-3)alkyl, heterocyclyl, amino,
(C.sub.1-6)alkylamino, di-(C.sub.1-6)alkylamino, where the alkyl
groups of di-(C.sub.1-6)alkylamino may be the same or different,
carboxy, aminocarbonyl, N--(C.sub.1-6alkyl)aminocarbonyl,
N,N-di-(C.sub.1-6alkyl)aminocarbonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminocarbonyl may be the same or different,
aminocarbonyl(C.sub.1-3)alkyl,
N--(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl,
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl, where the
(C.sub.1-6)alkyl groups of
N,N-di-(C.sub.1-6)alkylaminocarbonyl(C.sub.1-3)alkyl may be the
same or different, and aminosulfonyl; and
[0081] --X-- is --CH.dbd.CH--, --CH.sub.2CH.sub.2--, --NH--CO--,
--CONH-- or --(CH(OH)).sub.2--; or
[0082] pharmaceutically acceptable salts thereof.
[0083] A preferred embodiment of the present invention provides
compounds of Formula I wherein A is Cl, CH.sub.3 or CF.sub.3. In a
more preferred embodiment, A is Cl.
[0084] Another preferred embodiment of the present invention
provides compounds of Formula I wherein R1 is C.sub.1-6alkylamino,
C.sub.1-6alkoxy where C.sub.1-6alkoxy is substituted by
heterocyclyl, heterocyclyl, or R1 and R2, together with the phenyl
ring to which they are attached, form a five- to eight-membered
monocarbocyclic ring. In a more preferred embodiment, R1 is
C.sub.1-6alkoxy where C.sub.1-6alkoxy is substituted by
heterocyclyl, or heterocyclyl. In a most preferred embodiment, R1
is heterocyclyl.
[0085] Another preferred embodiment of the present invention
provides compounds of Formula I wherein R2 is H, or R1 and R2,
together with the phenyl ring to which they are attached, form a
five- to eight-membered monocarbocyclic ring. In a more preferred
embodiment R2 is H.
[0086] Another preferred embodiment of the present invention
provides compounds of Formula I wherein R3 is C.sub.1-6alkoxy or
H.
[0087] Another preferred embodiment of the present invention
provides compounds of Formula I wherein R4 is H, C.sub.1-6alkoxy,
N--(C.sub.1-6alkyl)aminocarbonyl, (C.sub.1-6alkyl)sulfonyl,
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino, where the
alkyl groups of
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino may be the
same or different, or heteroaryl. In a more preferred embodiment,
R4 is H, C.sub.1-6alkoxy, N--(C.sub.1-6alkyl)aminocarbonyl,
(C.sub.1-6alkyl)sulfonyl, or
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino, where the
alkyl groups of
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino may be the
same or different.
[0088] Another preferred embodiment of the present invention
provides compounds of Formula I wherein R5 is H,
N--(C.sub.1-6alkyl)aminosulfonyl, or
N,N-di-(C.sub.1-6alkyl)aminosulfonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminosulfonyl may be the same or
different.
[0089] In another preferred embodiment, --X-- is --CH.dbd.CH-- or
--CH.sub.2CH.sub.2--.
[0090] A particularly preferred embodiment of the present invention
provides compounds of Formula I wherein:
[0091] A is Cl, CH.sub.3 or CF.sub.3;
[0092] R1 is C.sub.1-6alkylamino, C.sub.1-6alkoxy where
C.sub.1-6alkoxy is substituted by heterocyclyl, heterocyclyl, or R1
and R2, together with the phenyl ring to which they are attached,
form a five- to eight-membered monocarbocyclic ring;
[0093] R2 is H, or R1 and R2, together with the phenyl ring to
which they are attached, form a five- to eight-membered
monocarbocyclic ring;
[0094] R3 is C.sub.1-6alkoxy or H;
[0095] R4 is H, C.sub.1-6alkoxy, N--(C.sub.1-6alkyl)aminocarbonyl,
(C.sub.1-6alkyl)sulfonyl,
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino, where the
alkyl groups of
N--[(C.sub.1-6alkyl)sulfonyl]-N--(C.sub.1-6alkyl)amino may be the
same or different, or heteroaryl;
[0096] R5 is H, N--(C.sub.1-6alkyl)aminosulfonyl, or
N,N-di-(C.sub.1-6alkyl)aminosulfonyl, where the alkyl groups of
N,N-di-(C.sub.1-6alkyl)aminosulfonyl may be the same or different;
and
[0097] X is --CH.dbd.CH-- or --CH.sub.2CH.sub.2--.
[0098] In other preferred embodiments, the present invention
provides any of the compounds as described in the Examples.
[0099] The present invention provides pharmaceutically acceptable
salts of compounds of Formula I. Pharmaceutically acceptable acid
addition salts of the compounds of Formula I include, but are not
limited to, salts derived from inorganic acids such as
hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydriodic,
and phosphorus, as well as the salts derived from organic acids,
such as aliphatic mono- and dicarboxylic acids, phenyl-substituted
alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic
acids, and aliphatic and aromatic sulfonic acids. Such salts thus
include, but are not limited to, sulfate, pyrosulfate, bisulfate,
sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate,
dihydrogenphosphate, metaphosphate, pyrophosphate, chloride,
bromide, iodide, acetate, propionate, caprylate, isobutyrate,
oxalate, malonate, succinate, suberate, sebacate, fumarate,
maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate,
dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate,
phenylacetate, citrate, lactate, maleate, tartrate, and
methanesulfonate. Also contemplated are the salts of amino acids
such as arginate, gluconate, galacturonate, and the like; see, for
example, Berge et al., "Pharmaceutical Salts," J. of Pharmaceutical
Science, 1977; 66:1-19.
[0100] The acid addition salts of the basic compounds may be
prepared by contacting the free base form with a sufficient amount
of the desired acid to produce the salt in the conventional manner.
The free base form may be regenerated by contacting the salt form
with a base and isolating the free base in the conventional manner.
The free base forms differ from their respective salt forms
somewhat in certain physical properties such as solubility in polar
solvents, but otherwise the salts are in general equivalent to
their respective free base for purposes of the present
invention.
[0101] Pharmaceutically acceptable base addition salts of compounds
of Formula I are formed with metals or amines, such as alkali and
alkaline earth metal hydroxides, or of organic amines. Examples of
metals used as cations include, but are not limited to, sodium,
potassium, magnesium, and calcium. Examples of suitable amines
include, but are not limited to, N,N'-dibenzylethylenediamine,
chloroprocaine, choline, diethanolamine, ethylenediamine
(ethane-1,2-diamine), N-methylglucamine, and procaine; see, for
example, Berge et al., supra., 1977.
[0102] The base addition salts of acidic compounds may be prepared
by contacting the free acid form with a sufficient amount of the
desired base to produce the salt in the conventional manner. The
free acid form may be regenerated by contacting the salt form with
an acid and isolating the free acid in a conventional manner. The
free acid forms differ from their respective salt forms somewhat in
certain physical properties such as solubility in polar solvents,
but otherwise the salts are in general equivalent to their
respective free acid for purposes of the present invention.
[0103] Some of the compounds in the present invention may exist as
stereoisomers, including enantiomers, diastereomers, and geometric
isomers. Geometric isomers include compounds of the present
invention that have alkenyl groups, which may exist as entgegen or
zusammen conformations, in which case all geometric forms thereof,
both entgegen and zusammen, cis and trans, and mixtures thereof,
are within the scope of the present invention. Some compounds of
the present invention have carbocyclyl groups, which may be
substituted at more than one carbon atom, in which case all
geometric forms thereof, both cis and trans, and mixtures thereof,
are within the scope of the present invention. All of these forms,
including (R), (S), epimers, diastereomers, cis, trans, syn, anti,
(E), (Z), tautomers, and mixtures thereof, are contemplated in the
compounds of the present invention.
[0104] The compounds to be used in the present invention can exist
in unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms, including hydrated forms,
are equivalent to unsolvated forms and are intended to be
encompassed within the scope of the present invention.
III. Pharmaceutical Compositions
[0105] The present invention further provides pharmaceutical
compositions comprising a compound of the present invention (e.g.,
a compound of Formula I or a pharmaceutically acceptable salt
thereof), together with a pharmaceutically acceptable carrier,
diluent, or excipient therefor. The pharmaceutical composition may
contain two or more compounds of the present invention (i.e., two
or more compounds of the present invention may be used together in
the pharmaceutical composition). Preferably, the pharmaceutical
composition contains a therapeutically effective amount of at least
one compound of the present invention. In another embodiment, these
compositions are useful in the treatment of an ALK-, FAK- or
JAK2-mediated disorder or condition. The compounds of the invention
can also be combined in a pharmaceutical composition that also
comprises compounds that are useful for the treatment of cancer or
another ALK-, FAK- or JAK2-mediated disorder.
[0106] A compound of the present invention can be formulated as a
pharmaceutical composition in the form of a syrup, an elixir, a
suspension, a powder, a granule, a tablet, a capsule, a lozenge, a
troche, an aqueous solution, a cream, an ointment, a lotion, a gel,
an emulsion, etc. Preferably, a compound of the present invention
will cause a decrease in symptoms or a disease indicia associated
with an ALK-, FAK- or JAK2-mediated disorder as measured
quantitatively or qualitatively.
[0107] For preparing a pharmaceutical composition from a compound
of the present invention, pharmaceutically acceptable carriers can
be either solid or liquid. Solid form preparations include powders,
tablets, pills, capsules, cachets, suppositories, and dispersible
granules. A solid carrier can be one or more substances which may
also act as diluents, flavoring agents, binders, preservatives,
tablet disintegrating agents, or an encapsulating material.
[0108] In powders, the carrier is a finely divided solid which is
in a mixture with the finely divided active component (i.e.,
compound of the present invention). In tablets, the active
component is mixed with the carrier having the necessary binding
properties in suitable proportions and compacted in the shape and
size desired.
[0109] The powders and tablets contain from 1% to 95% (w/w) of the
active compound (i.e., compound of the present invention). In
another embodiment, the active compound ranges from 5% to 70%
(w/w). Suitable carriers are magnesium carbonate, magnesium
stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin,
tragacanth, methylcellulose, sodium carboxymethylcellulose, a low
melting wax, cocoa butter, and the like. The term "preparation" is
intended to include the formulation of the active compound with
encapsulating material as a carrier providing a capsule in which
the active component with or without other carriers, is surrounded
by a carrier, which is thus in association with it. Similarly,
cachets and lozenges are included. Tablets, powders, capsules,
pills, cachets, and lozenges can be used as solid dosage forms
suitable for oral administration.
[0110] For preparing suppositories, a low melting wax, such as a
mixture of fatty acid glycerides or cocoa butter, is first melted
and the active component is dispersed homogeneously therein, as by
stirring. The molten homogeneous mixture is then poured into
convenient sized molds, allowed to cool, and thereby to
solidify.
[0111] Liquid form preparations include solutions, suspensions, and
emulsions, for example, water or water/propylene glycol solutions.
For parenteral injection, liquid preparations can be formulated in
solution in aqueous polyethylene glycol solution.
[0112] Aqueous solutions suitable for oral use can be prepared by
dissolving the active component in water and adding suitable
colorants, flavors, stabilizers, and thickening agents as desired.
Aqueous suspensions suitable for oral use can be made by dispersing
the finely divided active component in water with viscous material,
such as natural or synthetic gums, resins, methylcellulose, sodium
carboxymethylcellulose, and other well-known suspending agents.
[0113] Also included are solid form preparations which are intended
to be converted, shortly before use, to liquid form preparations
for oral administration. Such liquid forms include solutions,
suspensions, and emulsions. These preparations may contain, in
addition to the active component, colorants, flavors, stabilizers,
buffers, artificial and natural sweeteners, dispersants,
thickeners, solubilizing agents, and the like.
[0114] The pharmaceutical preparation is preferably in unit dosage
form. In such form the preparation is subdivided into unit doses
containing appropriate quantities of the active component. The unit
dosage form can be a packaged preparation, the package containing
discrete quantities of preparation, such as packeted tablets,
capsules, and powders in vials or ampoules. Also, the unit dosage
form can be a capsule, tablet, cachet, or lozenge itself, or it can
be the appropriate number of any of these in packaged form.
[0115] The quantity of active component in a unit dose preparation
may be varied or adjusted from 0.1 mg to 1000 mg, preferably 1.0 mg
to 100 mg, or from 1% to 95% (w/w) of a unit dose, according to the
particular application and the potency of the active component. The
composition can, if desired, also contain other compatible
therapeutic agents.
[0116] Pharmaceutically acceptable carriers are determined in part
by the particular composition being administered, as well as by the
particular method used to administer the composition. Accordingly,
there is a wide variety of suitable formulations of pharmaceutical
compositions of the present invention (see, e.g., Remington: The
Science and Practice of Pharmacy, 20th ed., Gennaro et al. Eds.,
Lippincott Williams and Wilkins, 2000).
[0117] A compound of the present invention, alone or in combination
with other suitable components, can be made into aerosol
formulations (i.e., they can be "nebulized") to be administered via
inhalation. Aerosol formulations can be placed into pressurized
acceptable propellants, such as dichlorodifluoromethane, propane,
nitrogen, and the like.
[0118] Formulations suitable for parenteral administration, such
as, for example, by intravenous, intramuscular, intradermal, and
subcutaneous routes, include aqueous and non-aqueous, isotonic
sterile injection solutions, which can contain antioxidants,
buffers, bacteriostats, and solutes that render the formulation
isotonic with the blood of the intended recipient, and aqueous and
nonaqueous sterile suspensions that can include suspending agents,
solubilizers, thickening agents, stabilizers, and preservatives. In
the practice of this invention, compositions can be administered,
for example, by intravenous infusion, orally, topically,
intraperitoneally, intravesically or intrathecally. The
formulations of compounds can be presented in unit-dose or
multi-dose sealed containers, such as ampoules and vials. Injection
solutions and suspensions can be prepared from sterile powders,
granules, and tablets of the kind previously described.
[0119] The dose administered to a subject, in the context of the
present invention, should be sufficient to effect a beneficial
therapeutic response in the subject over time. The dose will be
determined by the efficacy of the particular compound employed and
the condition of the subject, as well as the body weight or surface
area of the subject to be treated. The size of the dose also will
be determined by the existence, nature, and extent of any adverse
side-effects that accompany the administration of a particular
compound in a particular subject. In determining the effective
amount of the compound to be administered in the treatment or
prophylaxis of the disorder being treated, the physician can
evaluate factors such as the circulating plasma levels of the
compound, compound toxicities, and/or the progression of the
disease, etc. In general, the dose equivalent of a compound is from
about 1 .mu.g/kg to 10 mg/kg for a typical subject. Many different
administration methods are known to those of skill in the art.
[0120] For administration, compounds of the present invention can
be administered at a rate determined by factors that can include,
but are not limited to, the LD.sub.50 of the compound, the
pharmacokinetic profile of the compound, contraindicated drugs, and
the side-effects of the compound at various concentrations, as
applied to the mass and overall health of the subject.
Administration can be accomplished via single or divided doses.
IV. Methods of Treatment
[0121] In another aspect, the present invention provides a method
of treating a subject suffering from an ALK-, FAK- or JAK2-mediated
disorder or condition comprising: administering to the subject a
therapeutically effective amount of a compound of Formula I or a
pharmaceutically acceptable salt form thereof. In another aspect,
the present invention provides a compound of Formula I or a
pharmaceutically acceptable salt form thereof for use in treating a
subject suffering from an ALK-, FAK- or JAK2-mediated disorder or
condition. Preferably, the compound of Formula I or a
pharmaceutically acceptable salt form thereof is administered to
the subject in a pharmaceutical composition comprising a
pharmaceutically acceptable carrier. In another aspect, the present
invention provides a pharmaceutical composition comprising a
compound of Formula I or a pharmaceutically acceptable salt form
thereof for use in treating a subject suffering from an ALK-, FAK-
or JAK2-mediated disorder or condition. In another embodiment, the
ALK-, FAK- or JAK2-mediated condition or disorder is cancer. In
another embodiment, the ALK-, FAK- or JAK2-mediated condition is
selected from anaplastic large cell lymphoma, inflammatory
myofibroblastic tumor, glioblastoma, and other solid tumors. In
another embodiment, the ALK-, FAK- or JAK2-mediated condition is
selected from colon cancer, breast cancer, renal cancer, lung
cancer, hemangioma, squamous cell myeloid leukemia, melanoma,
glioblastoma, and astrocytoma.
[0122] The ALK-, FAK- or JAK2-mediated disorder or condition can be
treated prophylactically, acutely, and chronically using compounds
of the present invention, depending on the nature of the disorder
or condition. Typically, the host or subject in each of these
methods is human, although other mammals can also benefit from the
administration of a compound of the present invention.
[0123] In another embodiment, the present invention provides a
method of treating a proliferative disorder in a subject in need
thereof, comprising administering to the subject a therapeutically
effective amount of a compound of Formula I or a pharmaceutically
acceptable salt form thereof. In another aspect, the present
invention provides a compound of Formula I or a pharmaceutically
acceptable salt form thereof for use in treating a proliferative
disorder in a subject in need thereof. Preferably, the compound of
Formula I or a pharmaceutically acceptable salt form thereof is
administered to the subject in a pharmaceutical composition
comprising a pharmaceutically acceptable carrier. In another
aspect, the present invention provides a pharmaceutical composition
comprising a compound of Formula I or a pharmaceutically acceptable
salt form thereof for use in treating a proliferative disorder in a
subject. In certain embodiments, the proliferative disorder is
ALK-, FAK- or JAK2-mediated. In certain embodiments, the
proliferative disorder is cancer. In certain embodiments, the
proliferative disorder is selected from anaplastic large cell
lymphoma, inflammatory myofibroblastic tumor, glioblastoma, and
other solid tumors. In certain embodiments, the prolifereative
disorder is selected from colon cancer, breast cancer, renal
cancer, lung cancer, hemangioma, squamous cell myeloid leukemia,
melanoma, glioblastoma, and astrocytoma.
[0124] The proliferative disorder can be treated prophylactically,
acutely, and chronically using compounds of the present invention,
depending on the nature of the disorder or condition. Typically,
the host or subject in each of these methods is human, although
other mammals can also benefit from the administration of a
compound of the present invention.
[0125] In therapeutic applications, the compounds of the present
invention can be prepared and administered in a wide variety of
oral and parenteral dosage forms. Thus, the compounds of the
present invention can be administered by injection, that is,
intravenously, intramuscularly, intracutaneously, subcutaneously,
intraduodenally, or intraperitoneally. Also, the compounds
described herein can be administered by inhalation, for example,
intranasally. Additionally, the compounds of the present invention
can be administered transdermally. In another embodiment, the
compounds of the present invention are delivered orally. The
compounds can also be delivered rectally, bucally or by
insufflation.
[0126] The compounds utilized in the pharmaceutical method of the
invention can be administered at the initial dosage of about 0.001
mg/kg to about 100 mg/kg daily. In another embodiment, the daily
dose range is from about 0.1 mg/kg to about 10 mg/kg. The dosages,
however, may be varied depending upon the requirements of the
subject, the severity of the condition being treated, and the
compound being employed. Determination of the proper dosage for a
particular situation is within the skill of the practitioner.
Generally, treatment is initiated with smaller dosages which are
less than the optimum dose of the compound. Thereafter, the dosage
is increased by small increments until the optimum effect under the
circumstances is reached. For convenience, the total daily dosage
may be divided and administered in portions during the day, if
desired.
V. Chemistry
[0127] Unless otherwise indicated, all reagents and solvents were
obtained from commercial sources and used as received. .sup.1H NMRs
were obtained on a Bruker Avance at 400 MHz in the solvent
indicated with tetramethylsilane as an internal standard.
Analytical HPLC was run using a Zorbax RX-C8, 5.times.150 mm column
eluting with a mixture of acetonitrile and water containing 0.1%
trifluoroacetic acid with a gradient of 10-100%. LCMS results were
obtained from a Bruker Esquire 2000 Mass Spec with the Agilent 1100
HPLC equipped with an Agilent Eclipse XDB-C8, 2.times.30 mm 3.5
micron column. The column was at room temperature, with a run time
of five (5) minutes, a flow rate of 1.0 mL/min, and a solvent
mixture of 10% (0.1% formic acid/water):100% (acetonitrile/0.1%
formic acid). Automated normal phase column chromatography was
performed on a CombiFlash Companion (ISCO, Inc.). Reverse phase
preparative HPLC was performed on a Gilson GX-281 equipped with
Gilson 333 and 334 pumps using a Phenomenex 00F-4454-00-AX
Gemini-NX 5.mu. C18 column. Melting points were taken on a Mel-Temp
apparatus and are uncorrected.
Synthesis
[0128] The compounds of the present invention can be prepared in a
number of ways well known to one skilled in the art of organic
synthesis. The compounds of the present invention can be
synthesized using the methods described below as well as using
methods known to one skilled in the art of organic chemistry or
variations thereon as appreciated by those skilled in the art. The
preferred methods include, but are not limited to or by, those
described below. Unless otherwise stated, compounds are of
commercial origin or readily synthesized by standard methods well
known to one skilled in the art of organic synthesis.
[0129] The compounds of this invention may be prepared using the
reactions and techniques described in this section. The reactions
are performed in solvents appropriate to the reagents, and
materials employed are suitable for the transformations being
effected. Also, in the description of the synthetic methods below,
it is to be understood that all proposed reaction conditions,
including choice of solvent, reaction atmosphere, reaction
temperature, duration of experiment and workup procedures are
chosen to be conditions standard for that reaction which should be
readily recognized by one skilled in the art of organic
synthesis.
[0130] It is understood that the examples and embodiments described
herein are for illustrative purposes only and that various
modifications or changes in light thereof will be suggested to
persons skilled in the art and are to be included within the spirit
and purview of this application and the scope of the appended
claims. Specific chemical transformations are listed in the ensuing
schemes and one skilled in the art appreciates that a variety of
different reagents may be used in place of those listed. Common
replacements for such reagents can be found in, but not limited to,
texts such as "Encyclopedia of Reagents for Organic Synthesis" Leo
A. Paquette, John Wiley & Son Ltd (1995) or "Comprehensive
Organic Transformations: A Guide to Functional Group Preparations"
Richard C. Larock. Wiley-VCH and "Strategic Applications of Named
Reactions in Organic Synthesis" Kurti and Czako, Elsevier, 2005 and
references therein.
[0131] Compounds of the present invention, wherein X is
--CH.dbd.CH-- or --CH.sub.2CH.sub.2-- may be prepared according to
Scheme I.
##STR00004##
[0132] In Scheme I, R1', R2', R3', R4', R5' and A, are R1, R2, R3,
R4, R5 and A, respectively, as defined herein, or are synthetic
precursor moieties thereto, that may be converted to R1, R2, R3,
R4, R5 and A, respectively, by methods known in the art, or as
disclosed herein. In Scheme I, one of W or Y is a vinyl group, and
the other is bromo.
[0133] In Scheme I, an aniline is coupled with a chloropyrimidine,
in the presence of base, to form a phenylaminopyrimidine. This, in
turn, is coupled with another aniline, in an alcohol, in the
presence of an acid catalyst, to form a disubstituted pyrimidine.
The olefin-containing macrocycle is formed under Heck reaction
conditions, for example in the presence of palladium acetate,
tri-o-tolyl phosphine and triethyl amine. The macrocyclic olefin is
then reduced to afford the compound of Formula I wherein X is
--CH.sub.2--CH.sub.2--.
EXAMPLES
Example 1
6,13,19-Triaza-4-chloro-7,14.sup.1,3-dibenzena-1.sup.2,4-pyrimidinacyclodo-
cosaph-12-one
##STR00005##
[0134] Example 1a
3-(2,5-Dichloro-pyrimidin-4-ylamino)-benzoic acid methyl ester
[0135] A mixture of 2,4,5-Trichloro-pyrimidine (840 mg, 4.58 mmol),
3-amino-benzoic acid methyl ester (690 mg, 4.6 mmol), potassium
carbonate (K.sub.2CO.sub.3) (1.9 g, 14 mmol) in dimethylformamide
(DMF) (20 mL) were stirred at 80.degree. C. for 4 hours. After
cooling to room temperature, the mixture was treated with water and
the resulting solid was filtered and rinsed liberally with water.
After air drying there remained 770 mg of crude product
3-(2,5-Dichloro-pyrimidin-4-ylamino)-benzoic acid methyl ester,
which was used for the subsequent step without further
manipulation.
Example 1b
3-[2-(3-Amino-phenylamino)-5-chloro-pyrimidin-4-ylamino]-benzoic
acid methyl ester
[0136] 3-(2,5-Dichloro-pyrimidin-4-ylamino)-benzoic acid methyl
ester (435 mg, 1.46 mmol), m-phenylenediamine (100 mg, 1 mmol), and
4M hydrogen chloride (HCl) in 1,4-dioxane (300 mL) were combined in
methanol (10 mL) and warmed to reflux under an inert atmosphere for
5 hours. The resulting solid was filtered off, and the filtrate
concentrated under reduced pressure. The residue from the filtrate
was dissolved in dimethylsulfoxide (DMSO) and purified via
preparative HPLC. The desired fractions were partitioned between
saturated aqueous sodium bicarbonate (NaHCO.sub.3) and ethyl
acetate (EtOAc). The organic phase was dried over Na.sub.2SO.sub.4,
filtered, and concentrated under reduced pressure to yield 170 mg
(40%) of desired
3-[2-(3-Amino-phenylamino)-5-chloro-pyrimidin-4-ylamino]-benzoic
acid methyl ester, which was used without further manipulation. LC:
95%; LC/MS: M+H=370.1.
Example 1c
3-[2-(3-Amino-phenylamino)-5-chloro-pyrimidin-4-ylamino]-benzoic
acid
[0137] Lithium hydroxide (39 mg, 1.6 mmol) in water was added to a
mixture of
3-[2-(3-Amino-phenylamino)-5-chloro-pyrimidin-4-ylamino]-benzoic
acid methyl ester (120 mg, 0.32 mmol) in methanol (25 mL) and the
mixture was warmed at 55.degree. C. for 16 hours. The mixture was
concentrated under reduced pressure, then treated with water (2 mL)
followed by 1N HCl (1.7 mL). The mixture was concentrated under
reduced pressure and the residue was used for the subsequent step
without further manipulation. M+H=356.2.
Example 1d
6,13,19-Triaza-4-chloro-7,14.sup.1,3-dibenzena-1.sup.2,4-pyrimidinacyclodo-
cosaph-12-one
[0138] At room temperature
N-(3-Dimethylaminopropyl)-N'ethylcarbodiimide hydrochloride (130
mg, 0.67 mmol) was added to a mixture of
3-[2-(3-Amino-phenylamino)-5-chloro-pyrimidin-4-ylamino]-benzoic
acid (110 mg, 0.322 mmol) and hydroxybenzotriazole (65 mg, 0.48
mmol) in DMF (5 mL). After 16 hours the reaction mixture was
concentrated under reduced pressure and the residue purified by
Gilson reverse phase HPLC. Solid precipitated from the purified
fractions and was filtered to yield 19 mg of
6,13,19-Triaza-4-chloro-7,14.sup.1,3-dibenzena-1.sup.2,4-pyrimid-
inacyclodocosaph-12-one as a white solid. The filtrate was
lyophylized to yield a second 42 mg portion of
6,13,19-Triaza-4-chloro-7,14.sup.1,3-dibenzena-1.sup.2,4-pyrimidinacyclod-
ocosaph-12-one. LC: 100%; LC/MS: M+H=338.17; 1H NMR (DMSO-d6)
.delta. 9.82 (s, 1H, exchangeable), 9.28 (s, 1H, exchangeable),
9.25 (s, 1H, exchangeable), 8.07 (s, 1H), 7.96 (S, 1H), 7.76 (s,
1H), 7.25-7.21 (m, 1H), 7.09-7.00 (m, 3H), 6.73-6.71 (d, 1H),
6.68-6.66 (d, 1H).
Example 2
(14Z)-6-Chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14.-
3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decae-
ne
##STR00006##
[0139] Example 2a
N(2)-[3-Bromo-4-(4-methyl-piperain-1-yl)-phenyl]-5-chloro-N(4)-(3-vinyl-ph-
enyl)-pyrimidine-2,4-diamine
[0140]
N(2)-[3-Bromo-4-(4-methyl-piperain-1-yl)-phenyl]-5-chloro-N(4)-(3-v-
inyl-phenyl)-pyrimidine-2,4-diamine was prepared in a similar
manner as
N(2)-[3-Bromo-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-(2-methox-
y-5-vinyl-phenyl)-pyrimidine-2,4-diamine (prepared according to
Example 13d) after substituting
(2,5-Dichloro-pyrimidin-4-yl)-3-vinyl-phenyl)-amine for
(2,5-dichloro-pyrimidin-4-yl)-(2-methoxy-5-vinyl-phenyl)-amine and
after substituting 1-methoxy-2-propanol as the preferred solvent.
Also the reaction was warmed to 110.degree. C. and the workup
varied in that the resulting reaction solution was concentrated
under reduced pressure and the residue was partitioned between
CHCl.sub.3 and saturated aqueous NaHCO.sub.3. The organic phase was
dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure. The residue was triturated with CH.sub.3CN,
filtered, and rinsed with a small amount of CH.sub.3CN. The desired
product
N(2)-[3-Bromo-4-(4-methyl-piperain-1-yl)-phenyl]-5-chloro-N(4)-(3-vinyl-p-
henyl)-pyrimidine-2,4-diamine was isolated in 62% as a tan solid,
mp 149-153.degree. C. LC: 93%; LC/MS: M+H=501.0; .sup.1H NMR
(DMSO-d6) .delta. 9.35 (s, 1H), 8.85 (s, 1H), 8.16 (s, 1H), 7.83
(s, 1H), 7.70 (s, 1H), 7.61-7.58 (m, 2H), 7.37-7.33 (t, 1H),
7.28-7.26 (d, 1H), 6.97-6.95 (d, 1H), 6.75-6.68 (m, 1H), 5.81-5.77
(d, 1H), 5.27-5.24 (d, 1H), 2.87 (s, 4H), 2.46 (s, 4H), 2.23 (s,
3H).
Example 2b
(14Z)-6-Chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14.-
3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decae-
ne
[0141]
(14Z)-6-Chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracy-
clo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,1-
8-decaene was prepared in a similar manner as Methyl
(14Z)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene-10-carboxylate (prepared as in Example 16c) after substituting
N(2)-[3-Bromo-4-(4-methyl-piperain-1-yl)-phenyl]-5-chloro-N(4)-(3-vinyl-p-
henyl)-pyrimidine-2,4-diamine for
4-Bromo-2-(5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylamino]-p-
yrimidin-4-ylamino)-benzoic acid methyl ester. The desired product
(14Z)-6-Chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene was isolated in 75% yield as an off white solid. LC: 92%,
LC/MS: M+H=419.1.
[0142] A 25 mg sample of the 92% pure material was further purified
via preparative HPLC to return 17 mg of
(14Z)-6-Chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene as a TFA salt white lyophylate, which was the sample used for
biological testing. LC: 100%; LC/MS: M+H=419.1; .sup.1H NMR
(DMSO-d6) .delta. 9.58 (bs, 1H), 9.29 (s, 1H), 9.12 (s, 1H), 8.61
(s, 1H), 8.45-8.44 (d, 1H), 8.12 (s, 1H), 7.29-7.25 (t, 1H),
7.20-7.18 (d, 1H), 7.02-6.97 (m, 3H), 6.83-6.80 (d, 1H), 6.75-6.72
(d, 1H), 3.74 (bs, H.sub.2O), 3.54-3.51 (d, 2H), 3.29-3.22 (m, 4H),
2.93-2.89 (m, 5H).
Example 3
6-Chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.-
sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene
Trifluoroacetate (1:1)
##STR00007##
[0143]
6-Chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene
was prepared in a similar manner as
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)--
19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa--
1(20),3(22),4,6,9(21),10,12,16,18-nonaene (prepared according to
Example 5) after substituting
(14Z)-6-Chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene for
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpip-
erazin-1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup-
.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene. The
desired product
6-Chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[-
14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonae-
ne was isolated in 63% yield as a TFA salt, white lyophylate. LC:
99%, M+H=421.0; .sup.1H NMR (DMSO-d6) .delta. 9.7 (bs, 1H), 9.40
(s, 1H), 9.27 (s, 1H), 8.14 (s, 1H), 8.03-8.02 (d, 1H), 7.82 (s,
1H), 7.26-7.22 (t, 1H), 7.07-7.05 (d, 1H), 7.02-7.01 (d, 1H), 7.00
(s, 1H), 6.93-6.90 (m, 1H), 3.52-3.49 (d, 2H), 3.26-3.2 (m, 2H),
3.09-3.05 (d, 2H), 2.99-2.90 (m, 9H).
Example 4
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-
-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]do-
cosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene
##STR00008##
[0144] Example 4a
N-(4-Bromo-2-nitro-phenyl)-methanesulfonamide
[0145] Potassium nitrate (0.34 g, 3.36 mmol) was added neat to a
0.degree. C. solution of N-(4-bromo-phenyl)-methanesulfonamide
(0.75 g, 3.0 mmol) in acetonitrile (CH.sub.3CN) (20 mL) and
trifluoroacetic anhydride (2.6 mL). After 2 hours the 0.degree. C.
reaction mixture was slowly treated with saturated aqueous
NaHCO.sub.3 until the reaction mixture was basic. The resulting
solid was filtered and rinsed liberally with ice cold water. After
air drying there remained 0.75 g (85%) of desired
N-(4-Bromo-2-nitro-phenyl)-methanesulfonamide as a yellow solid,
which was used for the subsequent step without further
manipulation. LC: 98%, .sup.1H NMR (DMSO-d6) .delta. 9.88 (s, 1H),
8.23 (s, 1H), 7.95-7.93 (d, 1H), 7.59-7.57 (d, 1H), 3.152 (s,
3H).
Example 4b
N-(4-Bromo-2-nitro-phenyl)-N-methyl-methanesulfonamide
[0146] Methyl iodide (430 .mu.L, 6.9 mmol) was added to a mixture
of N-(4-Bromo-2-nitro-phenyl)-methanesulfonamide (0.68 g, 2.3 mmol)
and K.sub.2CO.sub.3 (1.3 g, 9.2 mmol) in DMF (10 mL). After 16
hours the reaction mixture was treated with water (20 mL). The
resulting solid was filtered, rinsed liberally with water, and air
dried to yield 0.68 g (96%) of desired
N-(4-Bromo-2-nitro-phenyl)-N-methyl-methanesulfonamide as a white
solid, which was used for the subsequent step without further
manipulation. LC: 100%, .sup.1H NMR (DMSO-d6) .delta. 8.23 (s, 1H),
8.02-8.00 (d, 1H), 7.80-7.77 (d, 1H), 3.26 (s, 3H), 3.06 (s,
3H).
Example 4c
N-Methyl-N-(2-nitro-4-vinyl-phenyl)-methanesulfonamide
[0147] N-Methyl-N-(2-nitro-4-vinyl-phenyl)-methanesulfonamide was
prepared in a similar manner as 2-Methoxy-5-vinyl-phenylamine
(prepared according to Example 13d) after substituting
N-(4-Bromo-2-nitro-phenyl)-N-methyl-methanesulfonamide for
5-bromo-2-methoxy-phenylamine. The workup was varied in that after
concentration of the reaction mixture, the residue was partitioned
between EtOAc (2.times.) and water. The combined organic phases
were dried over magnesium sulfate (MgSO.sub.4), filtered, and the
crude product in the filtrate was adsorbed directly onto silica gel
under reduced pressure. The residue was purified via normal phase
chromatography (EtOAc/Hexane eluent) to give desired
N-Methyl-N-(2-nitro-4-vinyl-phenyl)-methanesulfonamide in 69%
yield. LC: 98%; LC/MS: M+23=279.0.
Example 4d
N-(2-Amino-4-vinyl-phenyl)-N-methyl-methanesulfonamide
[0148] N-(2-Amino-4-vinyl-phenyl)-N-methyl-methanesulfonamide was
prepared in a similar manner as
4-(4-Methyl-piperazin-1-yl)-3-vinyl-phenylamine (prepared according
to Example 9e) after substituting
N-Methyl-N-(2-nitro-4-vinyl-phenyl)-methanesulfonamide for
1-Methyl-4-(4-nitro-2-vinyl-phenyl)-piperazine. The workup was
similar, except EtOAc was used for extractions rather than
chloroform (CHCl.sub.3). Compound
N-(2-Amino-4-vinyl-phenyl)-N-methyl-methanesulfonamide was
recovered in 99% yield as a brown oil and used directly after
workup for subsequent steps without further manipulation. LC: 97%;
LC/MS: M+23=249.0; .sup.1H NMR (CDCl.sub.3-d3) .delta. 7.11-7.09
(d, 1H), 6.85-6.83 (m, 2H), 6.66-6.59 (m, 1H), 5.74-5.70 (d, 1H),
5.29-5.27 (d, 1H), 4.24 (bs, 2H), 3.25 (s, 3H), 2.98 (s, 3H).
Example 4e
1-Bromo-2-fluoro-4-methoxy-5-nitro-benzene (10c) and
1-Bromo-4-fluoro-2-methoxy-5-nitro-benzene
[0149] 1-Bromo-2,4-difluoro-5-nitro-benzene (960 mg, 4.0 mmol) was
dissolved in MeOH (20 mL) was cooled to 0.degree. C. and treated
with 0.5M Sodium Methoxide in MeOH solution (8.1 mL, 4 mmol). After
2 hours the reaction solution was concentrated under reduced
pressure and the residue was partitioned between EtOAc and water.
The organic phase was dried over sodium sulfate (Na.sub.2SO.sub.4),
filtered, and concentrated under reduced pressure to give 833 mg
(83%) of yellowish oil. 1H NMR spectral analysis supported that the
product was a mixture of 1-Bromo-2-fluoro-4-methoxy-5-nitro-benzene
and 1-Bromo-4-fluoro-2-methoxy-5-nitro-benzene (.about.1:2 ratio).
The assignments of the respective regioisomers were determined
based on further analysis of a purified
1-Bromo-2-fluoro-4-methoxy-5-nitro-benzene analog, generated from
the subsequent steps to this reaction. .sup.1H NMR (DMSO-d6)
.delta. 8.41-8.39 (d, 0.63H), 8.36-8.34 (d, 0.26H), 7.57-7.54 (d,
0.30H), 7.45-7.42 (d, 0.70H), 4.01 (s, 2.2H), 3.95 (s, 0.9H).
Example 4f
1-(2-Bromo-5-methoxy-4-nitro-phenyl)-4-methyl-piperazine
[0150] The 833 mg mixture of
1-Bromo-2-fluoro-4-methoxy-5-nitro-benzene and
1-Bromo-4-fluoro-2-methoxy-5-nitro-benzene was treated at room
temperature with 1-methyl-piperazine (10 mL, 100 mmol). After 3
hours the mixture was diluted with water and then extracted with
EtOAc. The organic phase was dried over MgSO.sub.4, filtered, and
concentrated under reduced pressure to yield 1.13 g of oil. This
material was purified via preparative HPLC to separate
1-(2-Bromo-5-methoxy-4-nitro-phenyl)-4-methyl-piperazine from its
respective regioisomer. The purified fractions of
1-(2-Bromo-5-methoxy-4-nitro-phenyl)-4-methyl-piperazine from the
HPLC were partitioned between EtOAc and saturated aqueous
NaHCO.sub.3. The organic phase was dried over Na.sub.2SO.sub.4,
filtered, and concentrated under reduced pressure to give 0.26 g
(24%) of desired
1-(2-Bromo-5-methoxy-4-nitro-phenyl)-4-methyl-piperazine as an oil,
which crystallized upon sitting. Free basing of the purified
regioisomer
1-(4-Bromo-5-methoxy-2-nitro-phenyl)-4-methyl-piperazine, resulted
in a return of 0.56 g (51%) of this side product. Preliminary
analysis of
1-(2-Bromo-5-methoxy-4-nitro-phenyl)-4-methyl-piperazine: LC: 100%,
LC/MS: M+H=330.0.
Example 4g
5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine
[0151] 5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine
was prepared in a similar manner as
3-Bromo-4-(2-pyrrolidin-1-yl-ethoxy)-phenylamine (prepared
according to Example 22b) after substituting
1-(2-Bromo-5-methoxy-4-nitro-phenyl)-4-methyl-piperazine for
1-[3-(2-bromo-4-nitro-phenoxy)-propyl]-pyrrolidine. After workup a
mass return of 100% was returned, but .sup.1H NMR and LC/MS
confirmed the crude to be 80%
5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine, with a
20% impurity of the des-Br analog of
5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine. This
crude material was used for subsequent reactions without further
manipulation. It was also found that
5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine could be
prepared pure in 96% yield cleanly by following a procedure similar
to 4-(4-Methyl-piperazin-1-yl)-3-vinyl-phenylamine (prepared
according to Example 9e) after substituting
1-(2-Bromo-5-methoxy-4-nitro-phenyl)-4-methyl-piperazine for
1-Methyl-4-(4-nitro-2-vinyl-phenyl)-piperazine. LC/MS: M+H=302.0;
.sup.1H NMR (CDCl.sub.3-d3) .delta. 6.93 (s, 1H), 6.62 (s, 1H),
3.85 (s, 3H), 3.66 (s, 2H), 2.99 (bs, 4H), 2.61-2.59 (bm, 4H), 2.38
(s, 3H).
Example 4h
N-[2-(2,5-Dichloro-pyrimidin-4-ylamino)-4-vinyl-phenyl]-N-methyl-methanesu-
lfonamide
[0152]
N-[2-(2,5-Dichloro-pyrimidin-4-ylamino)-4-vinyl-phenyl]-N-methyl-me-
thanesulfonamide was prepared in a similar manner as
4-Bromo-2-(2,5-dichloro-pyrimidin-4-ylamino)-benzoic acid methyl
ester (prepared according to Example 16a) after substituting
N-(2-Amino-4-vinyl-phenyl)-N-methyl-methanesulfonamide for
2-Amino-4-bromo-benzoic acid methyl ester. The workup varied in
that after concentration of the reaction mixture the residue was
partitioned between EtOAc and water. The organic phase was dried
over Na.sub.2SO.sub.4, filtered, and the crude product in the
filtrate was adsorbed directly onto silica gel. The crude material
was purified via normal phase chromatography (EtOAc/hexane eluent)
to give desired
N-[2-(2,5-Dichloro-pyrimidin-4-ylamino)-4-vinyl-phenyl]-N-methyl-methanes-
ulfonamide in 50% yield as a yellow solid. LC: 93%; LC/MS:
M+H=373.2; .sup.1H NMR (DMSO-d6) .delta. 9.04 (s, 1H,
exchangeable), 8.45 (s, 1H), 7.93 (s, 1H), 7.64-7.62 (d, 1H),
7.47-7.45 (d, 1H), 6.82-6.75 (m, 1H), 5.91-5.87 (d, 1H), 5.40-5.37
(d, 1H), 3.17 (s, 3H), 3.04 (s, 3H).
Example 4i
N-(2-{2-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamino]-5-chlo-
ro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
[0153]
N-[2-(2,5-Dichloro-pyrimidin-4-ylamino)-4-vinyl-phenyl]-N-methyl-me-
thanesulfonamide (192 mg, 0.52 mmol), and
5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine (170 mg,
0.57 mmol), along with methanesulfonic acid (43.4 .mu.L, 0.67 mmol)
were combined in 2-methoxy-ethanol (3 mL) and warmed to 105.degree.
C. for 4 hours. The resulting solution was concentrated under
reduced pressure, dissolved in dimethyl sulfoxide (DMSO) and
purified via preparative HPLC. The purest fractions of desired
N-(2-{2-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamino]-5-chl-
oro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
were combined and partitioned between EtOAc (2.times.) and
saturated aqueous NaHCO.sub.3. The combined organic phases were
dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure. The 270 mg of yellow residue was triturated with
ice cold CH.sub.3CN, filtered, and rinsed with a small amount of
ice cold CH.sub.3CN to yield 110 mg (34%) of desired
N-(2-{2-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamino]-5-chl-
oro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
as an off white solid, which was used without further manipulation
for its subsequent cyclization to
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene. LC: 98%; LC/MS:
M+H=638.01. .sup.1H NMR (DMSO-d6) .delta. 8.41 (s, 1H,
exchangeable), 8.23 (s, 1H, exchangeable), 8.21 (s, 1H), 8.18 (s,
1H), 7.82 (s, 1H), 7.58-7.56 (d, 1H, J=8.25 Hz), 7.36-7.34 (d, 1H,
J=8.04 Hz), 6.81 (s, 1H), 6.61-6.54 (m, 1H), 5.77-5.72 (d, 1H,
J=17.69 Hz), 5.27-5.25 (d, 1H, J=10.77 Hz), 3.79 (s, 3H), 3.18 (s,
3H), 3.10 (s, 3H), 2.98 (s, 4H), 2.5 (s, 4H+DMSO), 2.24 (s,
3H).
Example 4j
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-
-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]do-
cosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene
[0154] Palladium(II) acetate (Pd(OAc).sub.2) (6.5 mg, 0.03 mmol)
was dissolved in CH.sub.3CN (3 mL), then tri-o-tolylphosphine (55
mg, 0.18 mmol) was added and the mixture was stirred at room
temperature for 15 minutes under argon (Ar).
N-(2-{2-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamino]-5-chl-
oro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
(90 mg, 0.14 mmol) and Et.sub.3N (300 .mu.L, 2 mmol) were added to
the resulting reaction mixture. The mixture was treated under
microwave irradiation at 120.degree. C. for 15 minutes. After
cooling the resulting solution was filtered through a Phenomenex
Strat-X-C cation catch and release resin, which was then rinsed
twice with CH.sub.3CN followed by methanol (MeOH). The resin
cartridge was transferred to a new filter flask, and the resin was
rinsed twice with 2M ammonia (NH.sub.3) in MeOH. The latter
methanol solution was concentrated under reduced pressure to give a
yellow residue. This material was treated with CH.sub.3CN (2 mL) to
yield a yellow solid, which was filtered and rinsed with a small
amount of ice cold CH.sub.3CN to give 57 mg (72%) of desired
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene as a yellow
solid, mp 278-280.degree. C. LC: 97%; LC/MS: M+H=556.1; .sup.1H NMR
(DMSO-d6) .delta. 8.90 (s, 1H), 8.37 (s, 1H), 8.21 (s, 1H), 8.18
(s, 1H; exchangeable), 8.02 (s, 1H; exchangeable), 7.57-7.55 (d,
1H, J=8.25 Hz), 7.15-7.13 (d, 1H, J=8.08 Hz), 6.78-6.75 (d, 1H,
J=13.0 Hz), 6.99 (s, 1H), 6.58-6.55 (d, 1H, J=13.1 Hz), 3.85 (s,
3H), 3.21 (s, 3H), 3.08 (s, 3H), 2.92 (bm, 4H), 2.5 (4H+DMSO), 2.25
(s, 3H).
Example 5
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)-1-
9-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1-
(20),3(22),4,6,9(21),10,12,16,18-nonaene Trifluoroacetate (1:1)
##STR00009##
[0156] At rt dipotassium azodicarboxylate (640 mg, 3.3 mmol) was
added to a solution of
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene (42 mg, 0.076
mmol) in pyridine (7 mL). The resulting mixture was treated with
acetic acid (0.53 mL, 9.24 mmol) and immediately warmed to
35.degree. C. After 24 hours the reaction mixture was treated with
an additional portion of dipotassium azodicarboxylate (270 mg, 1.4
mmol) and warming continued at 35.degree. C. for two days. The
mixture was then combined with water and after fizzing had ceased
the aqueous solution was partitioned between saturated aqueous
NaHCO.sub.3 and EtOAc. The organic phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced
pressure. The residue was purified via preparative HPLC. The
desired product
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)--
19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa--
1(20),3(22),4,6,9(21),10,12,16,18-nonaene was isolated in 74% yield
as a TFA salt, white lyophylate. LC: 99%; LC/MS: M+H=558.0; .sup.1H
NMR (DMSO-d6) .delta. 9.83 (bs, 1H, exchangeable), 8.32 (s, 1H,
exchangeable), 8.25 (s, 1H, exchangeable), 8.19 (s, 1H), 8.04 (s,
1H), 7.85 (s, 1H), 7.51-7.49 (d, 1H, J=8.21 Hz), 7.11-7.09 (d, 1H,
J=8.20 Hz), 6.72 (s, 1H), 3.81 (s, 3H), 3.54-3.51 (d, 2H),
3.28-3.21 (m, 2H), 3.18-3.14 (m, 5H), 3.04-2.92 (m, 8H).
Example 6
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-19-methoxy-2,-
4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4-
,6,9(21),10,12,16,18-nonaene Trifluoroacetate (1:1)
##STR00010##
[0157] Example 6a
4-Bromo-1-isopropylsulfanyl-2-nitro-benzene
[0158] 2-Propanethiol (0.76 mL, 8 mmol) was added to a rt solution
of 4-bromo-1-fluoro-2-nitro-benzene (1.10 g, 5.0 mmol) in ethanol
(EtOH) (5 mL) and the resulting solution was warmed at 50.degree.
C. for 4 days. After cooling to rt the solution was treated with
water (15 mL). The resulting yellow solid was filtered and rinsed
liberally with water. After air drying there remained 1.35 g (98%)
of desired 4-Bromo-1-isopropylsulfanyl-2-nitro-benzene as a yellow
solid. This material was used for the subsequent step without
further manipulation. LC: 96%; .sup.1H NMR (DMSO-d6) .delta.
8.30-8.29 (d, 1H), 7.90-7.87 (m, 1H), 7.67-7.64 (d, 1H), 3.76-3.73
(m, 1H), 1.30-1.28 (d, 6H).
Example 6b
4-Bromo-2-nitro-1-(propane-2-sulfonyl)-benzene
[0159] M-Chloroperbenzoic acid (1.18 g, 6.84 mmol) was added neat
to a 0.degree. C. solution of
4-Bromo-1-isopropylsulfanyl-2-nitro-benzene (0.55 g, 2.0 mmol) in
methylene chloride (CH.sub.2Cl.sub.2) (15 mL). The reaction was
warmed to room temperature (rt) and stirred for 16 hours. The
reaction solution was concentrated under reduced pressure and the
residual white solid was treated with saturated aqueous NaHCO.sub.3
(40 mL). After stirring for 10 minutes, the mixture was filtered
and the resulting solid was washed liberally with water. After air
drying there remained 0.56 g (91%) of desired
4-Bromo-2-nitro-1-(propane-2-sulfonyl)-benzene as a white solid.
This material was used for the subsequent step towards without
further manipulation. LC: 95%; .sup.1H NMR (DMSO-d6) .delta.
8.49-8.48 (d, 1H), 8.19-8.16 (m, 1H), 7.97-7.95 (d, 1H), 3.77-3.73
(m, 1H), 1.27-1.26 (d, 6H).
Example 6c
5-Bromo-2(propane-2-sulfonyl)-phenylamine
[0160] Iron (710 mg, 13 mmol) was added at rt to a stirring
solution of 4-Bromo-2-nitro-1-(propane-2-sulfonyl)-benzene (539 mg,
1.75 mmol) in tetrahydrofuran (THF) (10 mL) and acetic acid (14
mL). The mixture was warmed to 35.degree. C. and stirred 2 hours.
The mixture was then cooled to rt, filtered, and the resulting
solution was concentrated under reduced pressure. The residue was
partitioned between CHCl.sub.3 (2.times.) and saturated aqueous
NaHCO.sub.3. The combined organic phases were dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure
to yield 334 mg (69%) of 5-Bromo-2(propane-2-sulfonyl)-phenylamine
as a clear oil, which crystallized to a white solid upon sitting.
This material was used for its subsequent step without further
manipulation. LC: 97%; LC/MS: M+H=279.8.
Example 6d
2-Nitro-1-(propane-2-sulfonyl)-4-vinyl-benzene
[0161] 2-Nitro-1-(propane-2-sulfonyl)-4-vinyl-benzene was prepared
in a similar manner as 2-Methoxy-5-vinyl-phenylamine (prepared
according to Example 13b) after substituting
4-Bromo-2-nitro-1-(propane-2-sulfonyl)-benzene for
5-bromo-2-methoxy-phenylamine. The workup varied in that after
concentration of the reaction mixture, the residue was partitioned
between EtOAc (2.times.) and water. The combined organic phases
were dried over MgSO.sub.4, filtered, and the filtrate was adsorbed
directly onto silica gel under reduced pressure. The residue was
purified via normal phase chromatography (EtOAc/Hexane eluent) to
give desired 2-Nitro-1-(propane-2-sulfonyl)-4-vinyl-benzene in 55%
yield. LC: 99%; .sup.1H NMR (DMSO-d6) .delta. 8.26 (s, 1H),
8.02-7.98 (m, 2H), 6.95-6.87 (m, 1H), 6.27-6.23 (d, 1H), 5.67-5.64
(d, 1H), 3.78-3.70 (m, 1H), 1.28-1.26 (d, 6H).
Example 6e
2-(Propane-2-sulfonyl)-5-vinyl-phenylamine
[0162] Iron (391 mg, 7 mmol) was added at rt to a stirring solution
of 2-Nitro-1-(propane-2-sulfonyl)-4-vinyl-benzene (255 mg, 1 mmol)
in THF (5 mL) and acetic acid (10 mL). The mixture was warmed to
35.degree. C. and stirred 16 hours. The mixture was then cooled to
rt, filtered, and the resulting solution was concentrated under
reduced pressure. The residue was partitioned between EtOAc
(2.times.) and saturated aqueous NaHCO.sub.3. The combined organic
phases were dried over Na.sub.2SO.sub.4, filtered, and concentrated
under reduced pressure to yield 225 mg (99%) of
2-(Propane-2-sulfonyl)-5-vinyl-phenylamine as a clear oil. This
material was used for subsequent steps without further
manipulation. LC/MS: M+H=226.0; .sup.1H NMR (CDCl.sub.3-d3) .delta.
7.62-7.60 (d, 1H), 6.89-6.87 (d, 1H), 6.72 (s, 1H), 6.68-6.61 (m,
1H), 5.86-5.82 (d, 1H), 5.42-5.39 (d, 1H), 5.08 (bs, 2H), 3.37-3.30
(m, 1H), 1.34-1.32 (d, 6H).
Example 6f
(2,5-Dichloro-pyrimidin-4-yl)-[2-(propane-2-sulfonyl)-5-vinyl-phenyl]-amin-
e
[0163] 2-(Propane-2-sulfonyl)-5-vinyl-phenylamine (989 mg, 4.4
mmol), 2,4,5-trichloro-pyrimidine (7.02 mL, 61.2 mmol) and
ethyldiisopropylamine (EtN(i-Pr).sub.2) (1.2 mL, 7.2 mmol) were
warmed to 115.degree. C. for 16 hours. The reaction mixture was
concentrated under reduced pressure and the residue was partitioned
between CHCl.sub.3 (2.times.) and water. The organic phase was
dried over Na.sub.2SO.sub.4, filtered, and the crude product in the
filtrate was adsorbed directly onto silica gel. The crude material
was purified via normal phase chromatography (EtOAc/hexane eluent)
to give desired
(2,5-Dichloro-pyrimidin-4-yl)-[2-(propane-2-sulfonyl)-5-vinyl-phenyl]-ami-
ne in 51% yield as a yellow solid. LC: 100%; LC/MS: M+H=372.1;
.sup.1H NMR (DMSO-d6) .delta. 9.79 (s, 1H), 8.55-8.32 (d, 1H),
7.85-7.83 (bs, 1H), 6.89-6.82 (m, 1H), 6.07-6.03 (d, 1H), 5.57-5.54
(d, 1H), 3.52 (bm, 1H), 1.17-1.15 (d, 6H).
Example 6g
N(2)-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)--
[2-(propane-2-sulfonyl)-5-vinyl-phenyl]-pyrimidine-2,4-diamine
[0164]
(2,5-Dichloro-pyrimidin-4-yl)-[2-(propane-2-sulfonyl)-5-vinyl-pheny-
l]-amine (200 mg, 0.54 mmol), and
5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine (177 mg,
0.59 mmol), along with methanesulfonic acid (45.3 .mu.L, 0.70 mmol)
were combined in 2-methoxy-ethanol (3 mL) and warmed to 105.degree.
C. for 3 hours. The resulting solution was concentrated under
reduced pressure, dissolved in DMSO and purified via preparative
HPLC. The purest fractions of desired
N(2)-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-
-[2-(propane-2-sulfonyl)-5-vinyl-phenyl]-pyrimidine-2,4-diamine
were combined and partitioned between EtOAc (2.times.) and
saturated aqueous NaHCO.sub.3. The combined organic phases were
dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure. The 215 mg of yellow residue was triturated with
ice cold CH.sub.3CN, filtered, and rinsed with a small amount of
ice cold CH.sub.3CN to yield 70 mg (20%) of desired
N(2)-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-
-[2-(propane-2-sulfonyl)-5-vinyl-phenyl]-pyrimidine-2,4-diamine as
an off white solid, which was used without further manipulation for
its subsequent cyclization to
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-19-met-
hoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),-
3(22),4,6,9(21),10,12,14,16,18-decaene. LC: 99%; LC/MS: M+H=637.1;
.sup.1H NMR (DMSO-d6) .delta. 9.48 (s, 1H, exchangeable), 8.53 (s,
1H), 8.46 (s, 1H, exchangeable), 8.32 (s, 1H), 7.78-7.76 (m, 2H),
7.49-7.47 (d, 1H, J=7.64 Hz), 6.80 (s, 1H), 6.67-6.57 (m, 1H),
5.87-5.83 (d, 1H, J=17.72 Hz), 5.40-5.37 (d, 1H, J=10.85 Hz), 3.78
(s, 3H), 3.46-3.41 (m, 1H), 2.97 (s, 4H), 2.5 (s, 4H+DMSO), 2.24
(s, 3H), 1.18-1.16 (d, 6H, J=6.68 Hz).
Example 6h
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-19-meth-
oxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3-
(22),4,6,9(21),10,12,14,16,18-decaene
[0165] Pd(OAc).sub.2 (6.5 mg, 0.03 mmol) was dissolved in
CH.sub.3CN (3 mL), then tri-o-tolylphosphine (55 mg, 0.18 mmol) was
added and the mixture was stirred at rt for 15 minutes under Ar.
N(2)-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-
-[2-(propane-2-sulfonyl)-5-vinyl-phenyl]-pyrimidine-2,4-diamine (60
mg, 0.094 mmol) and Et.sub.3N (92 .mu.L, 0.66 mmol) were added to
the resulting reaction mixture. The mixture was treated under
microwave irradiation at 120.degree. C. for 15 minutes. After
cooling the resulting solution was filtered through a Phenomenex
Strat-X-C cation catch and release resin, which was then rinsed
twice with CH.sub.3CN followed by MeOH. The resin cartridge was
transferred to a new filter flask, and the resin was rinsed twice
with 2M NH.sub.3 in MeOH. The latter methanol solution was
concentrated under reduced pressure to give a yellow residue. This
material was treated with CH.sub.3CN (2 mL) to yield a yellow
solid, which was filtered and rinsed with a small amount of ice
cold CH.sub.3CN to give 33 mg (63%) of desired
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-19-met-
hoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),-
3(22),4,6,9(21),10,12,14,16,18-decaene as a yellow solid. LC: 97%;
LC/MS: M+H=555.1; .sup.1H NMR (DMSO-d6) .delta. 9.49 (s, 1H,
exchangeable), 9.27 (s, 1H), 8.42 (s, 1H), 8.25 (s, 1H), 8.16 (s,
1H, exchangeable), 7.74-7.72 (d, 1H, J=8.21 Hz), 7.28-7.26 (d, 1H,
8.00 Hz), 6.92-6.89 (d, 1H, J=13.08 Hz), 6.72 (s, 1H), 6.62-6.58
(d, 1H, J=13.20 Hz), 3.87 (s, 3H), 3.40-3.37 (m, 1H), 2.92 (s, 4H),
2.54 (s, 4H), 2.25 (s, 3H), 1.17-1.15 (d, 6H, J=6.76 Hz).
Example 6i
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-19-methoxy-2,-
4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4-
,6,9(21),10,12,16,18-nonaene Trifluoroacetate (1:1)
[0166]
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-19-met-
hoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),-
3(22),4,6,9(21),10,12,16,18-nonaene was prepared in a similar
manner as
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)--
19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa--
1(20),3(22),4,6,9(21),10,12,16,18-nonaene (prepared according to
Example 5) after substituting
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-19-met-
hoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),-
3(22),4,6,9(21),10,12,14,16,18-decaene for
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene. The desired
product
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-19-methoxy-2-
,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),-
4,6,9(21),10,12,16,18-nonaene was isolated in 53% yield as a TFA
salt white lyophylate. LC: 99%; LC/MS: M+H=557.10; .sup.1H NMR
(DMSO-d6) .delta. 9.71 (bs, 1H, exchangeable), 9.30 (s, 1H,
exchangeable), 8.38 (bs, 2H, 1 exchangeable), 7.86 (s, 1H),
7.70-7.68 (d, 1H, J=8.17 Hz), 7.22-7.20 (d, 1H, J=8.16 Hz), 6.73
(s, 1H), 3.81 (s, 3H), 3.54-3.51 (d, 2H), 3.40-3.33 (m, 1H),
3.29-3.21 (m, 2H), 3.19-3.15 (d, 2H), 3.05-2.91 (m, 9H), 1.13-1.11
(d, 6H, J=6.77 Hz).
Example 7
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(1-methyl-piperidin--
4-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene
##STR00011##
[0167] Example 7a
4-(2-Bromo-5-methoxy-phenyl)-piperidine
[0168] 4-(2-Bromo-5-methoxy-phenyl)-piperidine-1-carboxylic acid
tert-butyl ester (1.85 g, 5 mmol) in CH.sub.2Cl.sub.2 (6 mL) was
treated with trifluoroacetic acid (TFA) (2.5 mL, 32 mmol) at
0.degree. C. After 1 hour the mixture was warmed to rt and stirred
an additional 4 hours.
[0169] The mixture was then concentrated under reduced pressure and
the resulting residue partitioned between CHCl.sub.3 (2.times.) and
saturated aqueous NaHCO.sub.3. The combined organic phases were
dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure to yield desired
4-(2-Bromo-5-methoxy-phenyl)-piperidine in 100% yield as a tan oil,
which was used for the subsequent reaction without further
manipulation. LC: 95%.
Example 7b
4-(2-Bromo-5-methoxy-phenyl)-1-methyl-piperidine
[0170] A rt solution of 4-(2-Bromo-5-methoxy-phenyl)-piperidine
(220 mg, 0.80 mmol) in methanol (2 mL) and CH.sub.2Cl.sub.2 (3 mL)
was treated with 40% formaldehyde in water (0.13 mL, 1.6 mmol),
then acetic acid (54.6 uL, 0.96 mmol), followed by sodium
triacetoxyborohydride (305 mg, 1.44 mmol). After 5 hours the
mixture was treated with saturated aqueous NaHCO.sub.3. After
fizzing ceased, the reaction was diluted with additional
CH.sub.2Cl.sub.2 and the layers separated. The organic phase was
dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure to give desired
4-(2-Bromo-5-methoxy-phenyl)-1-methyl-piperidine as a clear oil in
98% yield. LC: 95%; LC/MS: M+H=286.0.
Example 7c
4-(2-Bromo-5-methoxy-phenyl)-1-methyl-piperidine nitrate
[0171] A solution of (2-Bromo-5-methoxy-phenyl)-1-methyl-piperidine
(970 mg, 3.4 mmol) in ether (Et.sub.2O) (2 mL) was added to a
premixed solution of fuming nitric acid (HNO.sub.3) (169 .mu.L) in
EtOH (2 mL). The resulting solution was cooled in an ice bath.
After a short time solid precipitated, which was filtered and
rinsed with Et.sub.2O yielding
4-(2-Bromo-5-methoxy-phenyl)-1-methyl-piperidine nitrate as a white
solid in 81% yield, mp 135-137.degree. C. LC: 96%; LC/MS:
M+H=286.0, .sup.1H NMR (DMSO-d6) .delta. 9.24 (bs, 1H), 7.55-7.53
(d, 1H), 6.85-6.83 (d, 1H), 5.78 (s, 1H), 3.77 (s, 3H), 3.53-3.50
(d, 2H), 3.18-3.07 (m, 3H), 2.82 (s, 3H), 2.00-1.97 (d, 2H),
1.85-1.76 (m, 2H).
Example 7d
4-(2-Bromo-5-methoxy-4-nitro-phenyl)-1-methyl-piperidine
[0172] A solution of
4-(2-Bromo-5-methoxy-phenyl)-1-methyl-piperidine nitrate (710 mg,
2.0 mmol) in water (2 mL) was added over 15 seconds to a 0.degree.
C. concentrated sulfuric acid (5 mL). After 10 minutes the reaction
mixture was diluted with ice (25 g), then while keeping cool
basified with 45% potassium hydroxide (KOH). The salts which
precipitated were filtered off and rinsed liberally with EtOAc. The
biphasic filtrate layers were separated, and the organic phase was
dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure to yield 670 mg of crude material. This material
was purified via preparative HPLC and the purest fractions of
desired product were combined and partitioned between EtOAc and
saturated aqueous NaHCO.sub.3. The organic phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure
to yield 320 mg (48%) of desired
4-(2-Bromo-5-methoxy-4-nitro-phenyl)-1-methyl-piperidine as a clear
oil. LC: 93%; LC/MS: 329.1; .sup.1H NMR (CDCl.sub.3-d3) .delta.
8.08 (s, 1H), 7.00 (s, 1H), 3.93 (s, 3H), 3.03-2.97 (m, 3H), 2.36
(s, 3H), 2.16-2.11 (m, 2H), 1.91-1.88 (d, 2H), 1.79-1.70 (m,
2H).
Example 7e
5-Bromo-2-methoxy-4-(1-methyl-piperidin-4-yl)-phenylamine
[0173] Iron (78 mg, 1.4 mmol) was added to a solution of
4-(2-Bromo-5-methoxy-4-nitro-phenyl)-1-methyl-piperidine (52 mg,
0.16 mmol) in THF (1 mL) and acetic acid (AcOH) (1 mL), and the
resulting mixture was warmed to 35.degree. C. for 5 hours, then at
rt for 2 days. The resulting solids were filtered off, rinsed with
THF, and the filtrate was concentrated under reduced pressure. The
residue was partitioned between EtOAc (2.times.) and saturated
aqueous NaHCO.sub.3. The combined organic phases were dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure
to yield 25 mg (53%) of desired
5-Bromo-2-methoxy-4-(1-methyl-piperidin-4-yl)-phenylamine as a
yellow oil. LC/MS: M+H=299.0; .sup.1H NMR (CDCl.sub.3-d3) .delta.
6.86 (s, 1H), 6.69 (s, 1H), 3.80 (s, 3H), 3.73 (bs, 2H), 2.98-2.95
(d, 2H), 2.89-2.83 (m, 1H), 2.33 (s, 3H), 2.12-2.06 (m, 2H),
1.84-1.81 (d, 2H), 1.75-1.66 (m, 2H).
Example 7f
N-(2-{2-[5-Bromo-2-methoxy-4-(1-methyl-piperidin-4-yl)-phenylamino]-5-chlo-
ro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
[0174]
N-(2-{2-[5-Bromo-2-methoxy-4-(1-methyl-piperidin-4-yl)-phenylamino]-
-5-chloro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
was prepared in a similar manner as
N-(2-{2-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamino]-5-chl-
oro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
(prepared according to Example 4i) after substituting
5-Bromo-2-methoxy-4-(1-methyl-piperidin-4-yl)-phenylamine for
5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine yielding
N-(2-{2-[5-Bromo-2-methoxy-4-(1-methyl-piperidin-4-yl)-phenylamino]-5-chl-
oro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
as a white solid in 52%, mp 196-198.degree. C. LC: 98%; LC/MS:
M+H=637.0.
Example 7g
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(1-methyl-piperidin--
4-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene
[0175]
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(1-methyl-pip-
eridin-4-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup-
.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene was
prepared in a similar manner as
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene (prepared
according to Example 4j) after substituting
N-(2-{2-[5-Bromo-2-methoxy-4-(1-methyl-piperidin-4-yl)-phenylamino]-5-chl-
oro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
for
N-(2-{2-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamino]-5-chl-
oro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
yielding
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(1-methyl--
piperidin-4-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.-
sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene as a
light tan solid in 62%, mp 244-246.degree. C. LC: 95%; LC/MS:
M+H=555.1; .sup.1H NMR (DMSO-d6) .delta. 8.7 (s, 1H), 8.23 (s, 1H,
exchangeable), 8.18 (s, 1H, exchangeable), 8.16 (s, 1H), 7.98 (s,
1H), 7.52-7.50 (d, 1H, J=7.18 Hz), 7.09-7.07 (d, 1H, J=8.52 Hz),
6.91-6.88 (d, 1H, J=13.16 Hz), 6.83 (s, 1H), 6.73-6.70 (d, 1H,
J=12.24 Hz), 3.81 (s, 3H), 3.18 (s, 3H), 3.06 (s, 3H), 2.88-2.85
(d, 2H), 2.73 (m, 1H), 2.20 (s, 3H), 2.01-1.95 (m, 2H), 1.71-1.63
(m, 4H).
Example 8
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(1-methyl-piperidin-4-yl)--
19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa--
1(20),3(22),4,6,9(21),10,12,16,18-nonaene Trifluoroacetate
(1:1)
##STR00012##
[0177]
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(1-methyl-piperidin-
-4-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]-
docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene was prepared in a
similar manner as
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpipera-
zin-1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,-
13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene (prepared
according to Example 5) after substituting
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(1-methyl-piperidin-
-4-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]-
docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene for
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene yielding
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(1-methyl-piperidin-4-yl)-
-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-
-1(20),3(22),4,6,9(21),10,12,16,18-nonaene as a TFA salt lyophylate
in 60% yield. LC: 99%; LC/MS: M+H=557.1; .sup.1H NMR (DMSO-d6)
.delta. 9.31 (bs, 1H, exchangeable), 8.26 (s, 1H, exchangeable),
8.22 (s, 1H, exchangeable), 8.17 (s, 1H), 8.12 (s, 1H), 7.71 (s,
1H), 7.48-7.46 (d, 1H, J=8.44 Hz), 7.09-7.07 (d, 1H, J=8.16 Hz),
6.69 (s, 1H), 3.84 (s, 3H), 3.7-3.5 (bm, 2H+H.sub.2O), 3.17 (s,
3H), 3.09-2.94 (bm, 10H), 2.84 (s, 3H), 1.91 (bm, 4H).
Example 9
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-
-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),-
3(22),4,6,9(21),10,12,14,16,18-decaene Trifluoroacetate (1:1)
##STR00013##
[0178] Example 9a
N-(2-Amino-4-bromo-phenyl)-N-methyl-methanesulfonamide
[0179] Iron (810 mg, 14 mmol) was added to a solution of
N-(4-Bromo-2-nitro-phenyl)-N-methyl-methanesulfonamide (618 mg, 2.0
mmol) in THF (10 mL) and AcOH (16 mL), and the resulting mixture
was warmed to 35.degree. C. for 5 hours. The resulting solids were
filtered off, rinsed with THF, and the filtrate was concentrated
under reduced pressure. The residue was partitioned between EtOAc
(2.times.) and saturated aqueous NaHCO.sub.3. The combined organic
phases were dried over Na.sub.2SO.sub.4, filtered, and concentrated
under reduced pressure to yield 550 mg (98%) of desired
N-(2-Amino-4-bromo-phenyl)-N-methyl-methanesulfonamide as a clear
oil, and used directly for subsequent steps without further
manipulation. LC: 93%; .sup.1H NMR (DMSO-d6) .delta. 7.14-7.12 (d,
1H), 6.92 (s, 1H), 6.69-6.67 (d, 1H), 5.43 (s, 2H), 3.05 (s,
6H).
Example 9b
N-[4-Bromo-2-(2,5-dichloro-pyrimidin-4-ylamino)-phenyl]-N-methyl-methanesu-
lfonamide (5 g)
[0180] N-(2-Amino-4-bromo-phenyl)-N-methyl-methanesulfonamide (550
mg, 2.0 mmol) and 2,4,5-trichloropyrimidine (678 .mu.L, 5.9 mmol)
were combined with EtN(i-Pr).sub.2 (940 .mu.L, 5.4 mmol) in
N-Methylpyrrolidinone (5 mL) and warmed to 100.degree. C. for 2
days. The resulting mixture was concentrated under high vacuum and
the residue treated with 1:1 H.sub.2O:CH.sub.3CN (8 mL). The
resulting solid was filtered and rinsed with ice cold 1:1
H.sub.2O:CH.sub.3CN (2 mL), then with MeOH, yielding 587 mg (70%)
of desired
N-[4-Bromo-2-(2,5-dichloro-pyrimidin-4-ylamino)-phenyl]-N-methyl-methanes-
ulfonamide as a tannish solid, which was used for subsequent steps
without further manipulation. LC: 95%; LC/MS: M+H=426.9.
Example 9c
1-(2-Bromo-4-nitro-phenyl)-4-methyl-piperazine
[0181] 1-(2-Bromo-4-nitro-phenyl)-4-methyl-piperazine was prepared
in a similar manner as
1-[3-(2-bromo-4-nitro-phenoxy)-propyl]-pyrrolidine (prepared
according to Example 22a) after substituting 1-methyl-piperazine
for N-.beta.-hydroxyethylpyrrolidine. In addition, no
K.sub.2CO.sub.3 was used, and the reaction was run at 90.degree. C.
rather than 100.degree. C. The desired product
1-(2-Bromo-4-nitro-phenyl)-4-methyl-piperazine was isolated in 97%
as a yellow solid. LC: 98%; .sup.1H NMR (DMSO-d6) .delta. 8.38-8.37
(d, 1H), 8.21-8.18 (m, 1H), 7.29-7.27 (d, 1H), 3.18-3.16 (bm, 4H),
2.50 (s, 4H+DMSO), 2.25 (s, 3H).
Example 9d
1-Methyl-4-(4-nitro-2-vinyl-phenyl)-piperazine
[0182] To a room temperature mixture of
1-(2-Bromo-4-nitro-phenyl)-4-methyl-piperazine (300 mg, 1.0 mmol),
K.sub.2CO.sub.3 (663 mg, 4.8 mmol), and
tetrakis(triphenylphosphine)palladium(0) (Pd(PPh.sub.3).sub.4) (200
mg, 0.2 mmol) in 1,2-dimethoxyethane (6 mL) and water (1 mL) was
added vinylboronic acid dibutyl ester (0.52 mL, 2.4 mmol). The
mixture was then warmed to 90.degree. C. for 5 h. The resulting
mixture was concentrated under reduced pressure, and the residue
was partitioned between aqueous 1N HCl and EtOAc. The aqueous phase
was basified with 8N KOH while cooling. The resulting solid was
filtered off, rinsed with ice cold water, and air dried yielding
1-Methyl-4-(4-nitro-2-vinyl-phenyl)-piperazine in 70% yield as a
yellow solid. LC: 98%; LC/MS M+H=248.1, .sup.1H NMR (DMSO-d6)
.delta. 8.24-8.23 (d, 1H), 8.12-8.09 (m, 1H), 7.18-7.16 (d, 1H),
6.82-6.75 (m, 1H), 5.97-5.93 (d, 1H), 5.46-5.43 (d, 1H), 3.08-3.06
(m, 4H), 2.50 (s, 4H+DMSO), 2.25 (s, 3H).
Example 9e
4-(4-Methyl-piperazin-1-yl)-3-vinyl-phenylamine
[0183] Iron (2.93 g, 52 mmol) was added at rt to a stirring
solution of 1-Methyl-4-(4-nitro-2-vinyl-phenyl)-piperazine (1.95 g,
7.9 mmol) in THF (30 mL) and acetic acid (60 mL). The mixture was
warmed to 35.degree. C. and stirred 16 hours. The mixture was then
cooled to rt, filtered, and the resulting solution was concentrated
under reduced pressure. The residue was partitioned between
CHCl.sub.3 (2.times.) and saturated aqueous NaHCO.sub.3. The
combined organic phases were dried over Na.sub.2SO.sub.4, filtered,
and concentrated under reduced pressure to yield 1.71 g (100%) of
4-(4-Methyl-piperazin-1-yl)-3-vinyl-phenylamine as a yellow tinted
oil, which crystallized upon sitting. This material was used for
its subsequent step without further manipulation. LC/MS:
M+H=218.1.
Example 9f
N-(4-Bromo-2-{5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylamino]-
-pyrimidin-4-ylamino}-phenyl)-N-methyl-methanesulfonamide
[0184]
N-[4-Bromo-2-(2,5-dichloro-pyrimidin-4-ylamino)-phenyl]-N-methyl-me-
thanesulfonamide (213 mg, 0.50 mmol), and
4-(4-Methyl-piperazin-1-yl)-3-vinyl-phenylamine (109 mg, 0.50
mmol), along with methanesulfonic acid (46 .mu.L, 0.71 mmol) were
combined in 2-methoxy-ethanol (6 mL) and warmed to 110.degree. C.
for 4 hours. The resulting solution was concentrated under reduced
pressure and the residue partitioned between EtOAc (2.times.) and
saturated aqueous NaHCO.sub.3. The combined organic phases were
dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure. The remaining brown oil was dissolved in
CH.sub.3CN (3 mL). The solid which crystallized was filtered and
rinsed with a small amount of ice cold CH.sub.3CN to yield 118 mg
(39%) of desired
N-(4-Bromo-2-{5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylamino-
]-pyrimidin-4-ylamino}-phenyl)-N-methyl-methanesulfonamide as an
off white solid, which was used without further manipulation for
its subsequent cyclization in the next step. LC: 90%; LC/MS:
M+H=608.1.
Example 9g
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-
-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),-
3(22),4,6,9(21),10,12,14,16,18-decaene Trifluoroacetate (1:1)
[0185] Pd(OAc).sub.2 (24 mg, 0.10 mmol), tri-o-tolylphosphine (140
mg, 0.45 mmol), and
N-(4-Bromo-2-{5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylamino-
]-pyrimidin-4-ylamino}-phenyl)-N-methyl-methanesulfonamide (80 mg,
0.13 mmol) were combined with CH.sub.3CN (2 mL) and triethylamine
(Et.sub.3N) (120 .mu.L, 0.9 mmol). The mixture was treated under
microwave irradiation at 120.degree. C. for 45 minutes. By LC
analysis, the reaction had not proceeded to completion, so the
reaction mixture was filtered and the solution recharged with
second portions of Pd(OAc).sub.2 (35 mg, 0.16 mmol),
tri-o-tolylphosphine (170 mg, 0.56 mmol), and Et.sub.3N (160 .mu.L,
1.1 mmol). The mixture was again treated under microwave
irradiation at 120.degree. C. for 120 minutes. After cooling, the
resulting mixture was filtered and the filtrate was concentrated
under reduced pressure. The concentrated residue was partitioned
between 1N HCl (1.4 mL) and EtOAc (2.times.). The aqueous phase was
purified directly via preparative HPLC and the purest fractions
combined and lyophilized to yield desired
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20)-
,3(22),4,6,9(21),10,12,14,16,18-decaene as its TFA salt in 37%
yield. This material was carried on for subsequent conversion
without further manipulation. LC: 100%; LC/MS: M+H=526.1; .sup.1H
NMR (DMSO-d6) .delta. 9.80 (bs, 1H), 9.42 (s, 1H), 8.89 (s, 1H),
8.42 (s, 1H), 8.23 (s, 1H), 8.20 (s, 1H), 7.58-7.56 (d, 1H),
7.20-7.18 (d, 1H), 7.07-7.01 (m, 2H), 6.88-6.84 (d, 1H), 6.74-6.71
(d, 1H), 3.53-3.51 (d, 2H), 3.35-3.21 (m, 7H), 3.09 (s, 3H),
2.97-2.91 (m, 2H), 2.89 (s, 3H).
Example 10
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)-2-
,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),-
4,6,9(21),10,12,16,18-nonaene Trifluoroacetate (1:1)
##STR00014##
[0187]
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20)-
,3(22),4,6,9(21),10,12,16,18-nonaene was prepared in a similar
manner as
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)--
19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa--
1(20),3(22),4,6,9(21),10,12,16,18-nonaene (prepared according to
Example 5) after substituting
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20)-
,3(22),4,6,9(21),10,12,14,16,18-decaene for
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene. The desired
product
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)--
2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22)-
,4,6,9(21),10,12,16,18-nonaene was isolated in 72% yield as a white
lyophylate. LC: 100% pure; LC/MS: M+H=528.1; .sup.1H NMR (DMSO-d6)
.delta. 9.58 (bs, 1H), 9.38 (s, 1H), 8.21 (s, 1H), 8.18 (s, 1H),
8.10 (s, 1H), 8.02 (s, 1H), 7.53-7.51 (d, 1H, J=8.09 Hz), 7.14-7.12
(d, 1H, J=8.12 Hz), 7.06-7.04 (d, 1H, J=8.43 Hz), 6.96-6.94 (d, 1H,
J=8.48 Hz), 3.53-3.50 (d, 2H), 3.25-3.22 (m, 2H), 3.19 (s, 3H),
3.11-2.91 (m, 14H).
Example 11
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,2-
2-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(-
21),10,12,14,16,18-decaene Trifluoroacetate (1:1)
##STR00015##
[0188] Example 11a
5-Bromo-2(propane-2-sulfonyl)-phenylamine
[0189] Iron (710 mg, 13 mmol) was added at rt to a stirring
solution of 4-Bromo-2-nitro-1-(propane-2-sulfonyl)-benzene (539 mg,
1.75 mmol) in THF (10 mL) and acetic acid (14 mL). The mixture was
warmed to 35.degree. C. and stirred 2 hours. The mixture was then
cooled to rt, filtered, and the resulting solution was concentrated
under reduced pressure. The residue was partitioned between
CHCl.sub.3 (2.times.) and saturated aqueous NaHCO.sub.3. The
combined organic phases were dried over Na.sub.2SO.sub.4, filtered,
and concentrated under reduced pressure to yield 334 mg (69%) of
5-Bromo-2(propane-2-sulfonyl)-phenylamine as a clear oil, which
crystallized to a white solid upon sitting. This material was used
for its subsequent step without further manipulation. LC: 97%;
LC/MS: M+H=279.8.
Example 11b
[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-(2,5-dichloro-pyrimidin-4-yl)-amin-
e
[0190] A mixture of 5-Bromo-2(propane-2-sulfonyl)-phenylamine (310
mg, 1.11 mmol) and 2,4,5-trichloropyrimidine (153 .mu.L, 1.34 mmol)
in DMF (2 mL) at 0.degree. C. was treated with neat sodium hydride
(NaH) (89 mg, 2.2 mmol; 60% oil dispersion) over 30 seconds. After
an hour at 0.degree. C., additional 2,4,5-trichloropyrimidine (150
.mu.L, 1.30 mmol) was added followed by neat NaH (160 mg, 4 mmol;
60% oil dispersion). After another hour the mixture was treated
with saturated aqueous NH.sub.4Cl, then diluted with water. The
resulting brownish solid was filtered and washed liberally with
water. The solid was then rinsed with ice cold CH.sub.3CN (3 mL).
The resulting 415 mg (88%) of tannish solid
[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-(2,5-dichloro-pyrimidin-4-yl)-ami-
ne was used for subsequent steps without further manipulation. LC:
100%; LC/MS: M+H=425.9.
Example 11c
N(4)-[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-5-chloro-N(2)-[4-(4-methyl-pi-
perazin-1-yl)-3-vinyl-phenyl]-pyrimidine-2,4-diamine
[0191]
[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-(2,5-dichloro-pyrimidin-4-y-
l)-amine (150 mg, 0.35 mmol), and
4-(4-Methyl-piperazin-1-yl)-3-vinyl-phenylamine (77 mg, 0.35 mmol),
along with methanesulfonic acid (33 .mu.L, 0.50 mmol) were combined
in 2-methoxy-ethanol (4 mL) and warmed to 110.degree. C. for 3
hours. The resulting solution was concentrated under reduced
pressure and the residue partitioned between CHCl.sub.3 (2.times.)
and saturated aqueous NaHCO.sub.3. The combined organic phases were
dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure. The 270 mg of remaining brown oil was dissolved
in CH.sub.3CN (3 mL). The solid which crystallized was filtered and
rinsed with a small amount of ice cold CH.sub.3CN to yield 90 mg
(40%) of desired
N(4)-[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-5-chloro-N(2)-[4-(4-methyl-p-
iperazin-1-yl)-3-vinyl-phenyl]-pyrimidine-2,4-diamine as a light
brown solid, which was used without further manipulation for its
subsequent cyclization to
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,-
22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9-
(21),10,12,14,16,18-decaene. LC: 92%; LC/MS: M+H=607.0.
Example 11d
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,2-
2-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(-
21),10,12,14,16,18-decaene Trifluoroacetate (1:1)
[0192] Pd(OAc).sub.2 (48 mg, 0.21 mmol), tri-o-tolylphosphine (275
mg, 0.90 mmol), and
N(4)-[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-5-chloro-N(2)-[4-(4-methyl-p-
iperazin-1-yl)-3-vinyl-phenyl]-pyrimidine-2,4-diamine (160 mg, 0.26
mmol) were combined with CH.sub.3CN (4 mL) and Et.sub.3N (250
.mu.L, 1.8 mmol). The mixture was treated under microwave
irradiation at 120.degree. C. for 120 minutes. After cooling, the
resulting mixture was filtered and the filtrate was concentrated
under reduced pressure. The concentrated residue was partitioned
between 1N HCl and EtOAc (2.times.). The aqueous phase was cooled
and neutralized with KOH solution until approximately pH 7, and
then diluted with saturated aqueous NaHCO.sub.3. The resulting
basic solution was extracted with CHCl.sub.3 (2.times.). The
combined CHCl.sub.3 extracts were dried over Na.sub.2SO.sub.4,
filtered, and concentrated under reduced pressure. The residue was
purified via preparative HPLC and the purest fractions combined and
lyophilized to yield desired
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,-
22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9-
(21),10,12,14,16,18-decaene as its TFA salt in 32% yield. This
material was carried on for subsequent conversion to
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22-tet-
raazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),1-
0,12,16,18-nonaene without further manipulation. LC: 100%; LC/MS:
M+H=525.1; .sup.1H NMR (DMSO-d6) .delta. 9.65 (bs, 1H), 9.55 (s,
1H), 9.48 (s, 1H), 9.26-9.25 (d, 1H), 8.46-8.45 (d, 1H), 8.28 (s,
1H), 7.77-7.75 (d, 1H), 7.33-7.31 (m, 1H), 7.12-7.05 (m, 2H),
7.02-6.98 (d, 1H), 6.77-6.74 (d, 1H), 4.1 (bs, H.sub.2O), 3.54-3.51
(d, 2H), 3.42-3.38 (m, 1H), 3.30-3.22 (m, 4H), 2.96-2.89 (m, 5H),
1.17-1.15 (d, 6H).
Example 12
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetr-
aazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10-
,12,16,18-nonaene
##STR00016##
[0194]
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,-
22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9-
(21),10,12,16,18-nonaene was prepared in a similar manner as
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)--
19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa--
1(20),3(22),4,6,9(21),10,12,16,18-nonaene (prepared according to
Example 5) after substituting
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,-
22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9-
(21),10,12,14,16,18-decaene for
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene. The workup
varied in that after the extractive workup the resulting crude
material was not purified via preparative HPLC, but rather via
crystallization from CH.sub.3CN. The desired product
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22-tet-
raazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),1-
0,12,16,18-nonaene was isolated in 50% yield as a yellow tinted
solid, mp>250.degree. C. LC: 98% pure; LC/MS: M+H=527.2; .sup.1H
NMR (DMSO-d6) .delta. 9.47 (s, 1H), 9.27 (s, 1H), 8.36-8.35 (d,
1H), 8.25 (s, 1H), 7.99-7.97 (d, 1H), 7.71-7.69 (d, 1H), 7.28-7.26
(m, 1H), 7.04-7.02 (d, 1H), 6.95-6.92 (m, 1H), 3.31 (bs,
1H+H.sub.2O), 3.04-2.98 (m, 4H), 2.81-2.79 (m, 4H), 2.5 (bs,
4H+DMSO), 2.25 (s, 3H), 1.14-1.12 (d, 6H).
Example 13
(14Z)-6-Chloro-10-methoxy-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatet-
racyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,-
16,18-decaene
##STR00017##
[0195] Example 13a
3-Bromo-4-(4-methyl-piperazin-1-yl)-phenylamine
[0196] 3-Bromo-4-(4-methyl-piperazin-1-yl)-phenylamine was prepared
in a similar manner as
3-Bromo-4-(2-pyrrolidin-1-yl-ethoxy)-phenylamine (prepared
according to Example 22b) after substituting
1-(2-Bromo-4-nitro-phenyl)-4-methyl-piperazine for
1-[3-(2-bromo-4-nitro-phenoxy)-propyl]-pyrrolidine. The crude
material was triturated with water to give
3-Bromo-4-(4-methyl-piperazin-1-yl)-phenylamine as a light brown
solid in 93% yield, which was used without further manipulation.
.sup.1H NMR (DMSO-d6) .delta. 6.90-6.88 (d, 1H), 6.82-6.81 (d, 1H),
6.54-6.51 (m, 1H), 5.03 (s, 2H), 2.79-2.77 (m, 4H), 2.43 (bm, 2H),
2.21 (s, 3H).
Example 13b
2-Methoxy-5-vinyl-phenylamine
[0197] To a room temperature mixture of
5-bromo-2-methoxy-phenylamine (610 mg, 3.0 mmol), K.sub.2CO.sub.3
(1.73 g, 12.5 mmol), and Pd(PPh.sub.3).sub.4(600 mg, 0.5 mmol) in
1,2-dimethoxyethane (18 mL) and water (3 mL) was added vinylboronic
acid dibutyl ester (2.37 mL, 10.8 mmol). The mixture was then
warmed to 90.degree. C. for 16 h. The resulting mixture was
concentrated under reduced pressure, and the residue was acidified
with aqueous 1N HCl. Attempted extraction with Et.sub.2O or EtOAc
yielded a solid between the biphasic mix, which was filtered off.
The aqueous phase of the filtrate was neutralized while cooling
with 30% NaOH, and the resulting mixture was washed with EtOAc
(2.times.). The latter EtOAc extracts were combined, dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure
to yield 320 mg (71%) of 2-Methoxy-5-vinyl-phenylamine as an oil,
which was used for subsequent steps without further manipulation.
.sup.1H NMR (DMSO-d6) .delta. 6.79-6.78 (d, 1H), 6.77-6.75 (d, 1H),
6.64-6.61 (m, 1H), 6.58-6.50 (m, 1H), 5.53-5.49 (d, 1H), 5.05-5.02
(d, 1H), 4.91 (bs, 2H), 3.75 (s, 3H).
Example 13c
(2,5-Dichloro-pyrimidin-4-yl)-(2-methoxy-5-vinyl-phenyl)-amine
[0198] 2-Methoxy-5-vinyl-phenylamine (970 mg, 6.5 mmol),
2,4,5-trichloro-pyrimidine (1.2 g, 6.5 mmol) and EtN(i-Pr).sub.2
(1.2 mL, 7.2 mmol) were combined in DMF (9 mL) at rt and stirred
for 72 hours. The reaction mixture was then diluted with ice cold
water (100 mL). The resulting solid was filtered and rinsed
liberally with water. The solid was further purified by trituration
with methanol (2.times.) to yield desired
(2,5-Dichloro-pyrimidin-4-yl)-(2-methoxy-5-vinyl-phenyl)-amine in
66% yield as an off white solid, LC: 97%; LC/MS: M+H=296.1; .sup.1H
NMR (DMSO-d6) .delta. 9.04 (s, 1H), 8.36 (s, 1H), 7.70 (s, 1H),
7.36-7.35 (d, 1H), 7.13-7.11 (d, 1H), 6.73-6.66 (m, 1H), 5.72-5.68
(d, 1H), 5.19-5.17 (d, 1H), 3.82 (s, 3H).
Example 13d
N(2)-[3-Bromo-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-(2-methoxy-
-5-vinyl-phenyl)-pyrimidine-2,4-diamine
[0199]
N(2)-[3-Bromo-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-(2--
methoxy-5-vinyl-phenyl)-pyrimidine-2,4-diamine was prepared in a
similar manner as
N(4)-[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-5-chloro-N(2)-[4-(-
4-methyl-piperazin-1-yl)-3-vinyl-phenyl]-pyrimidine-2,4-diamine
(prepared according to Example 11c) after substituting
3-Bromo-4-(4-methyl-piperazin-1-yl)-phenylamine for
4-(4-Methyl-piperazin-1-yl)-3-vinyl-phenylamine, substituting
(2,5-dichloro-pyrimidin-4-yl)-(2-methoxy-5-vinyl-phenyl)-amine for
[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-(2,5-dichloro-pyrimidin-4-yl)-ami-
ne, and after altering the solvent system from 2-methoxy-ethanol to
a 1:1 mixture of tert-butyl alcohol: 1,2-dimethoxyethane (120 mL of
solvent for a mmol reaction). The reaction mixture was warmed for 6
days at 85.degree. C. After concentrating the reaction mixture, the
crude mix was purified straightaway via preparative HPLC. The
purest fractions were then partitioned between EtOAc and saturated
aqueous NaHCO.sub.3. The organic phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure
to give desired
N(2)-[3-Bromo-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-(2-methox-
y-5-vinyl-phenyl)-pyrimidine-2,4-diamine in 20% yield. This
material was used without further manipulation for subsequent
macro-cyclization. LC: 93%; LC/MS: M+H=531.0.
Example 13e
(14Z)-6-Chloro-10-methoxy-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatet-
racyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,-
16,18-decaene
[0200]
(14Z)-6-Chloro-10-methoxy-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetr-
aazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10-
,12,14,16,18-decaene was prepared in a similar manner as
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene (prepared
according to Example 4j) after substituting
N(2)-[3-Bromo-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-(2-methox-
y-5-vinyl-phenyl)-pyrimidine-2,4-diamine for
N-[2-(2,5-Dichloro-pyrimidin-4-ylamino)-4-vinyl-phenyl]-N-methyl-methanes-
ulfonamide. The workup also varied in that desired product
(14Z)-6-Chloro-10-methoxy-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazate-
tracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14-
,16,18-decaene precipitated directly out of the reaction mixture.
The desired product was filtered, rinsed with a small amount of ice
cold CH.sub.3CN, and after air drying was isolated in 94% yield as
an off white solid, LC: 94%, M+H=449.1.
[0201] A small sample of the 94% pure material was further purified
via preparative HPLC to return pure
(14Z)-6-Chloro-10-methoxy-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazate-
tracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14-
,16,18-decaene as a TFA salt white lyophylate. LC: 99%; LC/MS:
M+H=449.1; .sup.1H NMR (DMSO-d6) .delta. 9.6 (bs, 1H), 9.38 (s,
1H), 9.04-9.03 (d, 1H), 8.41-8.40 (d, 1H), 8.19 (d, 1H), 7.84 (s,
1H), 7.11-7.01 (m, 4H), 6.68-6.60 (m, 2H), 3.90 (s, 3H), 3.5-3.2
(bm, 6H+H.sub.2O), 2.96-2.89 (m, 5H).
Example 14
6-Chloro-10-methoxy-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracycl-
o[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-non-
aene Trifluoroacetate (1:1)
##STR00018##
[0203]
6-Chloro-10-methoxy-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazate-
tracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16-
,18-nonaene was prepared in a similar manner as
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)--
19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa--
1(20),3(22),4,6,9(21),10,12,16,18-nonaene (prepared according to
Example 5) after substituting
(14Z)-6-Chloro-10-methoxy-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazate-
tracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14-
,16,18-decaene for
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene. The desired
product
6-Chloro-10-methoxy-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyc-
lo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-no-
naene was isolated in 41% yield as a TFA salt, white lyophylate.
LC: 99%; LC/MS: M+H=451.2; .sup.1H NMR (DMSO-d6) .delta. 9.6 (bs,
1H), 9.38 (s, 1H), 8.15 (s, 1H), 8.04 (bs, 1H), 7.95 (bs, 1H),
7.92-7.91 (d, 1H), 7.06-7.03 (d, 1H), 6.98-6.92 (m, 3H), 3.83 (s,
3H), 3.61 (bs, H.sub.2O), 3.53-3.50 (d, 2H), 3.25-3.22 (m, 2H),
3.10-3.07 (d, 2H), 2.99-2.96 (d, 2H), 2.92-2.87 (m, 7H).
Example 15
6-Chloro-17-(4-methylpiperazin-1-yl)-19-methoxy-2,4,8,22-tetraazatetracycl-
o[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-non-
aene Trifluoroacetate (1:1)
##STR00019##
[0204] Example 15a
(2,5-Dichloro-pyrimidin-4-yl)-3-vinyl-phenyl)-amine
[0205] 3-Vinyl-phenylamine (840 mg, 7.1 mmol),
2,4,5-trichloro-pyrimidine (1.3 g, 7.1 mmol) and EtN(i-Pr).sub.2
(1.23 mL, 7.1 mmol) were combined in DMF (10 mL) at rt and stirred
for 16 hours. The reaction mixture was then diluted with ice cold
water. The resulting white solid was filtered and rinsed liberally
with water. After air drying there remained 1.64 g (87%) of
(2,5-Dichloro-pyrimidin-4-yl)-3-vinyl-phenyl)-amine as a white
solid. LC: 97%; LC/MS: M+H=266.0.
Example 15b
N(2)-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)--
(3-vinyl-phenyl)-pyrimidine-2,4-diamine
[0206]
N(2)-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chlor-
o-N(4)-(3-vinyl-phenyl)-pyrimidine-2,4-diamine was prepared in a
similar manner as
N-(2-{2-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylam-
ino]-5-chloro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfona-
mide (prepared according to Example 4i) after substituting
(2,5-Dichloro-pyrimidin-4-yl)-3-vinyl-phenyl)-amine for
N-[2-(2,5-Dichloro-pyrimidin-4-ylamino)-4-vinyl-phenyl]-N-methyl-methanes-
ulfonamide, and heating the reaction for 8 hours rather than 4
hours to give desired
N(2)-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-
-(3-vinyl-phenyl)-pyrimidine-2,4-diamine as an oil in 10% yield.
The majority of
N(2)-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-
-(3-vinyl-phenyl)-pyrimidine-2,4-diamine was used without further
manipulation for its subsequent macro-cyclization. A small sample
of
N(2)-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-
-(3-vinyl-phenyl)-pyrimidine-2,4-diamine was crystallized from
CH.sub.3CN to yield it as a white solid, mp 126-129.degree. C.
LC/MS: M+H=451.1.
Example 15c
(14Z)-6-Chloro-17-(4-methylpiperazin-1-yl)-19-methoxy-2,4,8,22-tetraazatet-
racyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,-
16,18-decaene
[0207]
(14Z)-6-Chloro-17-(4-methylpiperazin-1-yl)-19-methoxy-2,4,8,22-tetr-
aazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10-
,12,14,16,18-decaene was prepared in a similar manner as
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene (prepared
according to Example 4j) after substituting
N(2)-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-
-(3-vinyl-phenyl)-pyrimidine-2,4-diamine for
N-[2-(2,5-Dichloro-pyrimidin-4-ylamino)-4-vinyl-phenyl]-N-methyl-methanes-
ulfonamide. The workup varied slightly in that the reaction mixture
was concentrated under reduced pressure. The residue was
partitioned between 1N HCl and EtOAc (2.times.). The aqueous phase
was neutralized, and extracted with CHCl.sub.3 (3.times.). The
CHCl.sub.3 extracts were combined, dried over Na.sub.2SO.sub.4,
filtered, and concentrated under reduced pressure to give crude
(14Z)-6-Chloro-17-(4-methylpiperazin-1-yl)-19-methoxy-2,4,8,22-tetraazate-
tracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14-
,16,18-decaene in 51% yield, which was carried on for subsequent
reduction towards
6-Chloro-17-(4-methylpiperazin-1-yl)-19-methoxy-2,4,8,22-tetraaza-
tetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,-
16,18-nonaene without further manipulation. LC/MS: M+H=449.0.
Example 15d
6-Chloro-17-(4-methylpiperazin-1-yl)-19-methoxy-2,4,8,22-tetraazatetracycl-
o[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-non-
aene Trifluoroacetate (1:1)
[0208]
6-Chloro-17-(4-methylpiperazin-1-yl)-19-methoxy-2,4,8,22-tetraazate-
tracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16-
,18-nonaene was prepared in a similar manner as
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)--
19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa--
1(20),3(22),4,6,9(21),10,12,16,18-nonaene (prepared according to
Example 5) after substituting
(14Z)-6-Chloro-17-(4-methylpiperazin-1-yl)-19-methoxy-2,4,8,22-tetraazate-
tracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14-
,16,18-decaene for
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene. The desired
product
6-Chloro-17-(4-methylpiperazin-1-yl)-19-methoxy-2,4,8,22-tetraazatetracyc-
lo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-no-
naene was isolated in 40% yield as a TFA salt, white lyophylate.
LC: 99%; LC/MS: M+H=451.1; .sup.1H NMR (DMSO-d6) .delta. 9.7 (bs,
1H), 9.36 (s, 1H), 8.14 (s, 1H), 8.12 (s, 1H), 7.90 (s, 1H), 7.76
(s, 1H), 7.25-7.21 (t, 1H), 7.05-7.03 (d, 1H), 7.00-6.98 (d, 1H),
6.69 (s, 1H), 3.81 (s, 3H), 3.70 (bs, water), 3.53 (d, 2H),
3.27-3.24 (m, 2H), 3.14-3.11 (d, 2H), 3.01-2.98 (d, 2H), 2.94 (s,
3H), 2.92-2.91 (d, 4H).
Example 16
(14Z)-6-Chloro-10-(N-methylcarboxamido)-17-(4-methylpiperazin-1-yl)-2,4,8,-
22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9-
(21),10,12,14,16,18-decaene
##STR00020##
[0209] Example 16a
4-Bromo-2-(2,5-dichloro-pyrimidin-4-ylamino)-benzoic acid methyl
ester
[0210] 2-Amino-4-bromo-benzoic acid methyl ester (1.2 g, 5.0 mmol)
was combined with 2,4,5-trichloro-pyrimidine (5.33 mL, 46.5 mmol)
and EtN(i-Pr).sub.2 (0.95 mL, 5.5 mmol), and the mixture was warmed
to 120.degree. C. After 16 hours the excess
2,4,5-trichloro-pyrimidine was removed under high vacuum, and the
residue was then treated with 1:1 MeOH:water. The resulting solid
was filtered, retriturated with 1:1 MeOH:water, refiltered, and the
resulting solid triturated in MeOH at 50.degree. C. After
filtering, the solid was rinsed liberally with MeOH. After air
drying 4-Bromo-2-(2,5-dichloro-pyrimidin-4-ylamino)-benzoic acid
methyl ester was recovered in 52% yield as a brown solid and was
carried on for subsequent steps without further manipulation.
LC/MS: M+H=377.9.
Example 16b
4-Bromo-2-(5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylamino]-py-
rimidin-4-ylamino)-benzoic acid methyl ester
[0211]
4-Bromo-2-(5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylam-
ino]-pyrimidin-4-ylamino)-benzoic acid methyl ester was prepared in
a similar manner as
N-(4-Bromo-2-{5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylamino-
]-pyrimidin-4-ylamino}-phenyl)-N-methyl-methanesulfonamide
(prepared according to Example 9f) after substituting
4-Bromo-2-(2,5-dichloro-pyrimidin-4-ylamino)-benzoic acid methyl
ester for
N-[4-Bromo-2-(2,5-dichloro-pyrimidin-4-ylamino)-phenyl]-N-methyl-meth-
anesulfonamide. The desired product
4-Bromo-2-(5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylamino]-p-
yrimidin-4-ylamino)-benzoic acid methyl ester was isolated in 52%
as a tan solid, LC: 98%; LC/MS: M+H=559.0
Example 16c
Methyl
(14Z)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracy-
clo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,1-
8-decaene-10-carboxylate
[0212] Methyl
(14Z)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene-10-carboxylate was prepared in a similar manner as
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene (prepared
according to Example 4j) after substituting
4-Bromo-2-(5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylamino]-p-
yrimidin-4-ylamino)-benzoic acid methyl ester for
N-[2-(2,5-Dichloro-pyrimidin-4-ylamino)-4-vinyl-phenyl]-N-methyl-methanes-
ulfonamide. The desired product Methyl
(14Z)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene-10-carboxylate was filtered directly from the reaction mixture,
rinsed with CH.sub.3CN and isolated in 54% yield as a bronze solid.
LC: 100%; LC/MS: M+H=477.1; .sup.1H NMR (DMSO-d6) .delta. 10.72 (s,
1H), 9.59 (s, 1H), 9.42 (s, 1H), 8.39 (s, 1H), 8.23 (s, 1H),
7.97-7.95 (d, 1H), 7.13-7.06 (m, 2H), 7.01-6.99 (d, 1H), 6.94-6.90
(d, 1H), 6.68-6.65 (d, 1H), 3.91 (s, 3H), 2.86 (s, 4H), 2.50 (s,
4H+DMSO), 2.25 (s, 3H)..quadrature.
Example 16d
(14Z)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14.-
3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decae-
ne-10-carboxylic acid
[0213] A solution of LiOH (11 mg, 0.46 mmol) in water (0.6 mL) was
added to a solution of Methyl
(14Z)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene-10-carboxylate (29 mg, 0.06 mmol) in MeOH (6 mL), and the
resulting mixture was warmed to 65.degree. C. for 16 hours. After
cooling to 0.degree. C., the mixture was treated with 1N HCl (0.46
mL, 0.46 mmol), then concentrated under reduced pressure and used
directly for the subsequent reaction towards
(14Z)-6-Chloro-10-(N-methylcarboxamido)-17-(4-methylpiperazin-1-yl)-2,4,8-
,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,-
9(21),10,12,14,16,18-decaene without further manipulation, LC:
100%, LC/MS: M+H=463.1.
Example 16e
(14Z)-6-Chloro-10-(N-methylcarboxamido)-17-(4-methylpiperazin-1-yl)-2,4,8,-
22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9-
(21),10,12,14,16,18-decaene
[0214] N-(3-Dimethylaminopropyl)-N'ethylcarbodiimide hydrochloride
(150 mg, 0.8 mmol) was added to a rt mixture of
(14Z)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene-10-carboxylic acid (28 mg, 0.06 mmol), 1-hydroxybenzotriazole
(11 mg, 0.084 mmol), methylammonium chloride (100 mg, 1 mmol), and
triethylamine (100 .mu.L, 0.7 mmol). After 7 days the reaction
mixture was partitioned between EtOAc (2.times.) and saturated
aqueous NaHCO.sub.3. The organic phases were combined, dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure
to yield 28 mg (98%) of
(14Z)-6-Chloro-10-(N-methylcarboxamido)-17-(4-methylpiperazin-1-yl)-2,4,8-
,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,-
9(21),10,12,14,16,18-decaene, which was used for subsequent step
without further manipulation. LC: 98%; LC/MS: M+H 476.2. A small
sample of the 98% pure material was further purified via
preparative HPLC to return pure
(14Z)-6-Chloro-10-(N-methylcarboxamido)-17-(4-methylpiperazin-1-yl)--
2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22)-
,4,6,9(21),10,12,14,16,18-decaene as a TFA salt white lyophylate.
LC: 100%; LC/MS: M+H=476.1; .sup.1H NMR (DMSO-d6) .delta. 11.55 (s,
1H), 9.55 (bs, 1H), 9.40 (s, 1H), 9.39 (s, 1H), 8.76-8.74 (m, 1H),
8.46-8.45 (d, 1H), 8.19 (s, 1H), 7.77-7.75 (d, 1H), 7.13-7.04 (m,
3H), 6.91-6.87 (d, 1H), 6.70-6.66 (d, 1H), 3.5 (d, 2H), 3.35-3.2
(bm, 4H+H.sub.2O), 2.95-2.89 (m, 5H), 2.81-2.80 (d, 3H).
Example 17
6-Chloro-10-(N-methylcarboxamido)-17-(4-methylpiperazin-1-yl)-2,4,8,22-tet-
raazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),1-
0,12,16,18-nonaene Trifluoroacetate (1:1)
##STR00021##
[0216]
6-Chloro-10-(N-methylcarboxamido)-17-(4-methylpiperazin-1-yl)-2,4,8-
,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,-
9(21),10,12,16,18-nonaene was prepared in a similar manner as
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)--
19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa--
1(20),3(22),4,6,9(21),10,12,16,18-nonaene (prepared according to
Example 5) after substituting
(14Z)-6-Chloro-10-(N-methylcarboxamido)-17-(4-methylpiperazin-1-yl)-2,4,8-
,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,-
9(21),10,12,14,16,18-decaene for
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene. The desired
product
6-Chloro-10-(N-methylcarboxamido)-17-(4-methylpiperazin-1-yl)-2,4,8,22-te-
traazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),-
10,12,16,18-nonaene was isolated in 20% yield as a TFA salt, white
lyophylate. LC: 100%; LC/MS: M+H=478.2; .sup.1H NMR (DMSO-d6)
.delta. 11.2 (s, 1H), 9.61 (bs, 1H), 9.38 (s, 1H), 8.66-8.65 (m,
1H), 8.40 (s, 1H), 8.18 (s, 1H), 8.08 (s, 1H), 7.70-7.67 (d, 1H),
7.08-6.96 (m, 3H), 3.53 (d, 2H), 3.26-3.23 (m, 2H), 3.12-3.09 (d,
2H), 3.0-2.93 (m, 6H), 2.91 (d, 3H), 2.79 (d, 3H).
Example 18
(14Z)-6-Chloro-10-(methanesulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22-t-
etraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21)-
,10,12,14,16,18-decaene Trifluoroacetate (1:1)
##STR00022##
[0217] Example 18a
(5-Bromo-2-methanesulfonyl-phenyl)-(2,5-dichloro-pyrimidin-4-yl)-amine
[0218]
[5-Bromo-2-methanesulfonyl)-phenyl]-(2,5-dichloro-pyrimidin-4-yl)-a-
mine was prepared in a similar manner as
[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-(2,5-dichloro-pyrimidin-4-yl)-ami-
ne (prepared according to Example 11a) after substituting
5-bromo-2-methanesulfonyl-phenylamine for
5-Bromo-2(propane-2-sulfonyl)-phenylamine.
(5-Bromo-2-methanesulfonyl-phenyl)-(2,5-dichloro-pyrimidin-4-yl)-amine
was isolated in 98% yield as a yellow solid. LC: 97%; LC/MS:
M+H=397.9.
Example 18b
N(4)-(5-Bromo-2-methanesulfonyl-phenyl)-5-chloro-N(2)-[4-(4-methyl-piperaz-
in-1-yl)-3-vinyl-phenyl]-pyrimidine-2,4-diamine
[0219]
N(4)-(5-Bromo-2-methanesulfonyl-phenyl)-5-chloro-N(2)-[4-(4-methyl--
piperazin-1-yl)-3-vinyl-phenyl]-pyrimidine-2,4-diamine was prepared
in a similar manner as
N(4)-[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-5-chloro-N(2)-[4-(4-methyl-p-
iperazin-1-yl)-3-vinyl-phenyl]-pyrimidine-2,4-diamine (prepared
according to Example 11c) after substituting
(5-Bromo-2-methanesulfonyl-phenyl)-(2,5-dichloro-pyrimidin-4-yl)-amine
for
[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-(2,5-dichloro-pyrimidin-4-yl)-
-amine.
N(4)-(5-Bromo-2-methanesulfonyl-phenyl)-5-chloro-N(2)-[4-(4-methyl-
-piperazin-1-yl)-3-vinyl-phenyl]-pyrimidine-2,4-diamine. LC: 92%;
LC/MS: M+H=579.0.
Example 18c
(14Z)-6-Chloro-10-(methanesulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22-t-
etraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21)-
,10,12,14,16,18-decaene Trifluoroacetate (1:1)
[0220]
(14Z)-6-Chloro-10-(methanesulfonyl)-17-(4-methylpiperazin-1-yl)-2,4-
,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,-
6,9(21),10,12,14,16,18-decaene was prepared in a similar manner as
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,-
22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9-
(21),10,12,14,16,18-decaene (prepared according to Example 11d)
after substituting
N(4)-[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-5-chloro-N(2)-[4-(4-methyl-p-
iperazin-1-yl)-3-vinyl-phenyl]-pyrimidine-2,4-diamine for
N(4)-[5-Bromo-2-(propane-2-sulfonyl)-phenyl]-5-chloro-N(2)-[4-(4-methyl-p-
iperazin-1-yl)-3-vinyl-phenyl]-pyrimidine-2,4-diamine.
(14Z)-6-Chloro-10-(methanesulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22--
tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21-
),10,12,14,16,18-decaene was isolated as a TFA salt, yellow
lyophylate in 20% yield. LC: 100%; LC/MS: M+H=497.1.
Example 19
6-Chloro-10-(methanesulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraaz-
atetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12-
,16,18-nonaene
##STR00023##
[0222]
6-Chloro-10-(methanesulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22--
tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21-
),10,12,16,18-nonaene was prepared in a similar manner as
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22-tet-
raazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),1-
0,12,16,18-nonaene (prepared according to Example 12) after
substituting
(14Z)-6-Chloro-10-(methanesulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22--
tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21-
),10,12,14,16,18-decaene for
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,-
22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9-
(21),10,12,14,16,18-decaene. The desired product
6-Chloro-10-(methanesulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraa-
zatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,1-
2,16,18-nonaene was isolated in 20% yield as a yellow tinted solid,
mp>250.degree. C. LC: 97%; LC/MS: M+H=499.1; .sup.1H NMR
(DMSO-d6) .delta. 9.41 (s, 1H), 9.08 (s, 1H), 8.35 (s, 1H), 8.24
(s, 1H), 7.98-7.97 (d, 1H), 7.77-7.75 (d, 1H), 7.27-7.25 (d, 1H),
7.03-7.01 (d, 1H), 6.93-6.90 (m, 1H), 3.21 (s, 3H), 3.08-2.99 (m,
4H), 2.80-2.78 (m, 4H), 2.50 (bm, 4H+DMSO), 2.25 (s, 3H).
Example 20
(14Z)-6-Chloro-10-methoxy-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo[-
14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-de-
caene
##STR00024##
[0223] Example 20a
N(2)-[3-Bromo-4-morpholin-4-yl-phenyl]-5-chloro-N(4)-(2-methoxy-5-vinyl-ph-
enyl)-pyrimidine-2,4-diamine
[0224]
N(2)-[3-Bromo-4-morpholin-4-yl-phenyl]-5-chloro-N(4)-(2-methoxy-5-v-
inyl-phenyl)-pyrimidine-2,4-diamine was prepared in a similar
manner as
N(2)-[3-Bromo-4-(4-methyl-piperazin-1-yl)-phenyl]-5-chloro-N(4)-(2-methox-
y-5-vinyl-phenyl)-pyrimidine-2,4-diamine (prepared according to
Example 13d) after substituting
3-Bromo-4-morpholin-4-yl-phenylamine for
3-Bromo-4-(4-methyl-piperazin-1-yl)-phenylamine and after altering
the solvent system from 2-methoxy-ethanol to a 1:1 mixture of
tert-butyl alcohol: 1,2-dimethoxyethane (120 mL of solvent for a
mmol reaction). The reaction mixture was warmed for 6 days at
85.degree. C., followed by the standard workup using EtOAc for the
extractive workup. Some desired
N(2)-[3-Bromo-4-morpholin-4-yl-phenyl]-5-chloro-N(4)-(2-methoxy-5-vinyl-p-
henyl)-pyrimidine-2,4-diamine crystallized relatively pure from
EtOAc on concentrating the filtrate, the remainder was purified via
normal phase chromatography to yield the desired product
N(2)-[3-Bromo-4-morpholin-4-yl-phenyl]-5-chloro-N(4)-(2-methoxy-5-vinyl-p-
henyl)-pyrimidine-2,4-diamine in 55% overall yield as a yellow
tinted solid, mp 186-190.degree. C. LC: 100%; LC/MS: M+H=518.0;
.sup.1H NMR (DMSO-d6) .delta. 9.36 (s, 1H), 8.23 (s, 1H), 8.16 (s,
1H), 7.95 (s, 1H), 7.8 (s, 1H), 7.58-7.57 (m, 1H), 7.33-7.32 (m,
1H), 7.13-7.11 (d, 1H), 6.96-6.94 (d, 1H), 6.7-6.6 (m, 1H),
5.67-5.62 (d, 1H), 5.13-5.10 (d, 1H), 3.85 (s, 3H), 3.74-3.71 (m,
4H), 2.88-2.85 (m, 4H).
Example 20b
(14Z)-6-Chloro-10-methoxy-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo[-
14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-de-
caene
[0225]
(14Z)-6-Chloro-10-methoxy-17-(morpholin-4-yl)-2,4,8,22-tetraazatetr-
acyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,1-
6,18-decaene was prepared in a similar manner as was Methyl
(14Z)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene-10-carboxylate (prepared according to Example 16c) after
substituting
N(2)-[3-Bromo-4-morpholin-4-yl-phenyl]-5-chloro-N(4)-(2-methoxy-5-vinyl-p-
henyl)-pyrimidine-2,4-diamine for
4-Bromo-2-(5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylamino]-p-
yrimidin-4-ylamino)-benzoic acid methyl ester. The desired product
(14Z)-6-Chloro-10-methoxy-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo-
[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-d-
ecaene was isolated in 53% yield as a gray solid, LC: 87%, LC/MS:
M+H=436.2.
Example 21
6-Chloro-10-methoxy-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo[14.3.1-
.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene
Trifluoroacetate (1:1)
##STR00025##
[0227]
6-Chloro-10-methoxy-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo-
[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nona-
ene was prepared in a similar manner as
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22-tet-
raazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),1-
0,12,16,18-nonaene (prepared according to Example 12) after
substituting
(14Z)-6-Chloro-10-methoxy-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo-
[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-d-
ecaene for
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1--
yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3-
(22),4,6,9(21),10,12,14,16,18-decaene. The desired product
6-Chloro-10-methoxy-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo[14.3.-
1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene
was isolated in 19% yield as a TFA salt, white lyophylate. LC: 99%;
LC/MS: M+H=438.22; .sup.1H NMR (DMSO-d6) .delta. 9.38 (bs, 1H),
8.20 (bs, 1H), 8.16 (s, 1H), 7.93 (s, 1H), 7.87-7.86 (d, 1H),
7.05-6.90 (m, 4H), 3.84 (s, 3H), 3.76-3.74 (m, 4H), 3.5 (bs, H2O),
2.96-2.89 (M, 4H), 2.80-2.78 (m, 4H).
Example 22
(14Z)-6-Chloro-17-[2-(pyrrolidin-1-yl)ethoxy]-2,4,8,22-tetraazatetracyclo[-
14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-de-
caene
##STR00026##
[0228] Example 22a
1-[3-(2-bromo-4-nitro-phenoxy)-propyl]-pyrrolidine
[0229] A mixture of 2-Bromo-1-fluoro-4-nitro-benzene (1.10 g, 5.0
mmol), N-.beta.-hydroxyethylpyrrolidine (6.0 mL, 52 mmol), and
K.sub.2CO.sub.3 (1.38 g, 10 mmol) was warmed to 100.degree. C. for
24 h. The mixture was cooled to rt and diluted with water. The
resulting solid was filtered, rinsed with water, and air dried to
yield 1.32 g (84%) of
1-[3-(2-bromo-4-nitro-phenoxy)-propyl]-pyrrolidine as a copper
colored solid. LC: 93%; .sup.1H NMR (DMSO-d6) .delta. 8.43-8.42 (d,
1H), 8.28-8.25 (m, 1H), 7.36-7.34 (d, 1H), 4.36-4.31 (t, 2H),
2.88-2.86 (t, 2H), 2.58-2.55 (m, 4H), 1.70-1.67 (m, 4H).
Example 22b
3-Bromo-4-(2-pyrrolidin-1-yl-ethoxy)-phenylamine
[0230] 1-[3-(2-bromo-4-nitro-phenoxy)-propyl]-pyrrolidine (0.74 g,
2.3 mmol) was combined with RaNi (120 mg) in ethanol (EtOH) (50 mL)
and the mixture was warmed to reflux. Hydrazine hydrate (1.0 mL, 2
mmol) was added dropwise to the refluxing solution and the elevated
temperature was maintained for 1 h. After cooling, the reaction
mixture was filtered, and the filtrate concentrated under reduced
pressure to yield 0.58 g (87%) of
3-Bromo-4-(2-pyrrolidin-1-yl-ethoxy)-phenylamine as a brown oil,
which was used without further manipulation. LC/MS: M+H=285.30.
Example 22c
N(2)-[3-Bromo-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5-chloro-N(4)-(3-vinyl--
phenyl)-pyrimidine-2,4-diamine
[0231]
N(2)-[3-Bromo-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5-chloro-N(4)-(3-
-vinyl-phenyl)-pyrimidine-2,4-diamine was prepared in a similar
manner as
N(2)-[3-Bromo-4-(4-methyl-piperain-1-yl)-phenyl]-5-chloro-N(4)-(3-vinyl-p-
henyl)-pyrimidine-2,4-diamine (prepared according to Example 2a)
after substituting 3-Bromo-4-(2-pyrrolidin-1-yl-ethoxy)-phenylamine
for 3-Bromo-4-(4-methyl-piperazin-1-yl)-phenylamine. The desired
product
N(2)-[3-Bromo-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5-chloro-N(4)-(3-vinyl-
-phenyl)-pyrimidine-2,4-diamine was isolated in 52% as white solid,
M+H=515.4
Example 22d
(14Z)-6-Chloro-17-[2-(pyrrolidin-1-yl)ethoxy]-2,4,8,22-tetraazatetracyclo[-
14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-de-
caene
[0232]
(14Z)-6-Chloro-17-[2-(pyrrolidin-1-yl)ethoxy]-2,4,8,22-tetraazatetr-
acyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,1-
6,18-decaene was prepared in a similar manner as Methyl
(14Z)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene-10-carboxylate (prepared according to Example 16c) after
substituting
N(2)-[3-Bromo-4-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-5-chloro-N(4)-(3-vinyl-
-phenyl)-pyrimidine-2,4-diamine for
4-Bromo-2-(5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylamino]-p-
yrimidin-4-ylamino)-benzoic acid methyl ester. The desired product
(14Z)-6-Chloro-17-[2-(pyrrolidin-1-yl)ethoxy]-2,4,8,22-tetraazatetracyclo-
[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-d-
ecaene was isolated in 62% yield as an copper colored solid. LC:
100%; LC/MS: M+H=434.1; .sup.1H NMR (DMSO-d6) .delta. 9.7 (bs, 1H),
9.24 (s, 1H), 9.11 (s, 1H), 8.63 (s, 1H), 8.51 (d, 1H), 8.12 (s,
1H), 7.29-7.25 (t, 1H), 7.21-7.19 (d, 1H), 7.04-6.99 (m, 2H),
6.96-6.94 (d, 1H), 6.89-6.86 (d, 1H), 6.73-6.70 (d, 1H), 4.28-4.26
(m, 2H), 3.64-3.56 (bm, 2H+water), 3.18 (m, 2H), 2.07 (bm, 2H),
1.91-1.89 (m, 2H).
Example 23
6-Chloro-17-[2-(pyrrolidin-1-yl)ethoxy]-2,4,8,22-tetraazatetracyclo[14.3.1-
.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene
Trifluoroacetate (1:1)
##STR00027##
[0234]
6-Chloro-17-[2-(pyrrolidin-1-yl)ethoxy]-2,4,8,22-tetraazatetracyclo-
[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nona-
ene was prepared in a similar manner as
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22-tet-
raazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),1-
0,12,16,18-nonaene (prepared according to Example 12) after
substituting
(14Z)-6-Chloro-17-[2-(pyrrolidin-1-yl)ethoxy]-2,4,8,22-tetraazatetracyclo-
[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-d-
ecaene for
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1--
yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3-
(22),4,6,9(21),10,12,14,16,18-decaene. The workup varied slightly
in that after the addition of water no solid precipitated, so the
aqueous solution was partitioned between saturated aqueous
NaHCO.sub.3 and EtOAc. The organic phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced
pressure. The residue was purified via preparative HPLC. The
desired product
6-Chloro-17-[2-(pyrrolidin-1-yl)ethoxy]-2,4,8,22-tetraazatetracyclo[14.3.-
1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene
was isolated as a TFA salt, white lyophylate in 40% yield. LC:
100%; LC/MS: M+H=436.1; .sup.1H NMR (DMSO-d6) .delta. 9.7 (bs, 1H),
9.16 (s, 1H), 9.05 (s, 1H), 8.09 (s, 1H), 7.82 (s, 1H), 7.80 (s,
1H), 7.23-7.19 (t, 1H), 7.07-7.05 (d, 1H), 6.97-6.95 (d, 1H), 6.88
(m, 2H), 4.25-4.23 (m, 2H), 3.64 (bs, 2H+H.sub.2O), 3.20 (m, 2H),
2.91 (s, 4H), 2.08 (m, 2H), 1.91 (m, 2H).
Example 24
(14Z)-6-Chloro-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.su-
p.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene
##STR00028##
[0235] Example 24a
N(2)-(3-Bromo-4-morpholin-4-yl-phenyl)-5-chloro-N(4)-3-vinyl-phenyl)-pyrim-
idine-2,4-diamine
[0236] (2,5-Dichloro-pyrimidin-4-yl)-3-vinyl-phenyl)-amine (270 mg,
1.0 mmol), 3-bromo-4-morpholin-4-yl-phenylamine (260 mg, 1.0 mmol),
and 4.0 M HCl in dioxane (280 .mu.L, 1.10 mmol) were combined in
2-methoxy-ethanol (11 mL) and warmed to 110.degree. C. for 16
hours. The resulting solution was concentrated under reduced
pressure and the residue was partitioned between ethyl acetate and
saturated aqueous NaHCO.sub.3. The organic phase was dried over
Na.sub.2SO.sub.4, filtered, and concentrated under reduced
pressure. The residue was triturated with CH.sub.3CN, filtered, and
rinsed with a small amount of CH.sub.3CN to yield
N(2)-(3-Bromo-4-morpholin-4-yl-phenyl)-5-chloro-N(4)-3-vinyl-phenyl)-pyri-
midine-2,4-diamine in 71% as white solid, mp 142-144.degree. C. LC:
91%; LC/MS: M+H=406.1; .sup.1H NMR (DMSO-d6) .delta. 9.37 (s, 1H),
8.86 (s, 1H), 8.16 (s, 1H), 7.85 (s, 1H), 7.70 (s, 1H), 7.63-7.60
(m, 2H), 7.38-7.34 (t, 1H), 7.28-7.26 (d, 1H), 6.99-6.96 (d, 1H),
6.76-6.68 (m, 1H), 5.81-5.77 (d, 1H), 5.27-5.24 (d, 1H), 3.73-3.71
(m, 4H), 2.88-2.86 (m, 4H).
Example 24b
(14Z)-6-Chloro-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.su-
p.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene
[0237] Palladium acetate (Pd(OAc).sub.2) (35 mg, 0.16 mmol),
tri-o-tolylphosphine (220 mg, 0.71 mmol), and
N(2)-(3-Bromo-4-morpholin-4-yl-phenyl)-5-chloro-N(4)-3-vinyl-phenyl)-pyri-
midine-2,4-diamine (150 mg, 0.31 mmol) were combined with
CH.sub.3CN (4 mL) and triethylamine (Et.sub.3N) (300 .mu.L, 2
mmol). The mixture was treated under microwave irradiation at
120.degree. C. for 30 minutes. After cooling, the resulting solid
was filtered and rinsed with ice cold CH.sub.3CN. After the
CH.sub.3CN trituration, the sample was also triturated with ethyl
ether (Et.sub.2O). The desired product
(14Z)-6-Chloro-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.s-
up.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene
was isolated in 43% yield as an off white solid, mp 267-270.degree.
C. LC: 95%; LC/MS: M+H=406.1; .sup.1H NMR (DMSO-d6) .delta. 9.18
(s, 1H), 9.03 (s, 1H), 8.65 (s, 1H), 8.44-8.43 (d, 1H), 8.10 (s,
1H), 7.27-7.23 (t, 1H), 7.18-7.16 (d, 1H), 7.01-6.97 (m, 2H),
6.95-6.93 (d, 1H), 6.85-6.82 (d, 1H), 6.72-6.68 (d, 1H), 3.76-3.75
(m, 4H), 2.84 (m, 4H).
Example 25
6-Chloro-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.-
1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene
##STR00029##
[0239] At rt dipotassium azodicarboxylate (784 mg, 4.0 mmol) was
added to a solution of
(14Z)-6-Chloro-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.s-
up.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene
(70 mg, 0.172 mmol) in pyridine (10 mL). The resulting mixture was
treated with acetic acid (AcOH) (0.56 mL, 9.8 mmol). After 3 days
the reaction mixture was treated with additional portions of
dipotassium azodicarboxylate (330 mg, 1.7 mmol) and AcOH (0.28 mL,
4.9 mmol) and warmed to 35.degree. C. for two days (LC supported
reaction >95% complete). The mixture was then combined with
water and the resulting solid was collected. This solid was
dissolved in DMSO and purified via preparative HPLC to yield 28 mg
(31%) of desired
6-Chloro-17-(morpholin-4-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7-
.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene as a
white lyophylate. LC: 100%; LC/MS: M+H=408.2; .sup.1H NMR (DMSO-d6)
.delta. 9.47 (bs, 2H), 8.16 (s, 1H), 7.95-7.94 (d, 1H), 7.80 (s,
1H), 7.25-7.21 (t, 1H), 7.05-7.00 (m, 3H), 6.91-6.88 (m, 1H),
3.76-3.74 (m, 4H), 3.02-2.96 (m, 4H), 2.78-2.77 (m, 4H).
Example 26
(14R,15S)-6-chloro-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13-
]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene-14,15-diol
##STR00030##
[0241] 4.8 M N-Methylmorpholine N-oxide in water (300 .mu.L, 1
mmol) was added to a solution of
(14Z)-6-Chloro-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]do-
cosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene (110 mg, 0.34
mmol) in THF (15 mL) and then a solution of potassium osmate,
dihydrate (25 mg, 0.068 mmol) in water (1 mL) was added and the
mixture was warmed to 55.degree. C. After 1.5 h, added acetone (5
mL) and additional water (1 mL), then heating was continued an
additional 2 hours. The reaction mixture was concentrated under
reduced pressure, the residue dissolved in 1000 uL of DMSO,
filtered and purified via preparative reverse phase HPLC. The front
running material was lyophilized to yield 6 mg (5%) of desired
product
(14R,15S)-6-chloro-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,1-
3]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene-14,15-diol as a
white lyophylate. LC: 98%; LC/MS: M+H=355.1; NMR (DMSO-d6) .delta.
8.12 (s, 1H), 7.79 (s, 1H), 7.73 (s, 1H), 7.29-7.08 (m, 5H),
6.99-6.97 (d, 1H), 4.80 (s, 1H), 4.5 (s, 1H).
Example 27
(14R,15S)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo-
[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nona-
ene-14,15-diol
##STR00031##
[0243] A room temperature solution of
(14Z)-6-Chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene (65 mg, 0.16 mmol) in pyridine (3 mL) was treated with a 0.2 M
aqueous solution of osmium tetraoxide (1.04 mL, 0.163 mmol). After
4.5 h the mixture was treated with a 0.5 M aqueous solution of
sodium sulfite (1 mL, 0.50 mmol), which was stirred for 5 minutes.
The mixture was extracted twice with CHCl.sub.3. The organic phases
were combined, dried over Na.sub.2SO.sub.4, filtered, and
concentrated under reduced pressure. The residue was purified via
preparative reverse phase HPLC to yield 4 mg (6%) of desired
(14R,15S)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracycl-
o[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-non-
aene-14,15-diol as a brown lyophylate. LC: 100%; LC/MS: M+H=453.1;
NMR (CDCl.sub.3/MeOH-d4) .delta. 8.06 (s, 1H), 7.99-7.98 (d, 1H),
7.82 (bs, 1H), 7.46-7.44 (d, 1H), 7.38-7.34 (m, 1H), 7.22-7.20 (d,
1H), 7.12-7.11 (d, 1H), 7.04-7.02 (m, 1H), 5.6 (s, 1H), 4.85
(2H+H.sub.2O), 3.60-3.43 (m, 6H), 3.13-3.10 (m, 2H), 3.01 (s,
3H).
Example 28
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(piperidin-4-yl)-19-methox-
y-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(2-
2),4,6,9(21),10,12,16,18-nonaene Trifluoroacetate (1:1)
##STR00032##
[0245] Under an inert atmosphere at rt
6-chloro-10-[2-methanesulfonyl-methyl-amino]-19-methoxy-17-(1-methyl-pipe-
ridin-4-yl)-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-
-1(20),3(22),4,6,9(21),10,12,16,18-nonaene (8 mg, 0.014 mmol) in
CHCl.sub.3 (3 mL) was treated with N,N-diisopropylethylamine (32
.mu.L, 0.19 mmol) followed by .alpha.-chloroethyl chloroformate (18
.mu.L, 0.17 mmol). The resulting mixture was warmed to 60.degree.
C. for 1 hour, and then concentrated under reduced pressure. The
resulting residue was treated with methanol (5 mL) and warmed to
50.degree. C. for 1 hour. The reaction mixture was again
concentrated under reduced pressure; the residue was dissolved in
DMSO and purified by preparative HPLC yielding the desired
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(piperidin-4-yl)-19-metho-
xy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(-
22),4,6,9(21),10,12,16,18-nonaene as a TFA salt, white lyophylate
(58%). LC: 97%; LC/MS: M+H=543.1; .sup.1H NMR (DMSO-d6) .delta.
8.59-8.56 (bd, 1H, exchangeable), 8.28-8.24 (bm, 3H, exchangeable),
8.17 (s, 1H), 8.11 (s, 1H), 7.70 (s, 1H), 7.48-7.46 (d, 1H, J=8.36
Hz), 7.08-7.06 (d, 1H, J=8.25 Hz), 6.71 (s, 1H), 3.79 (s, 3H),
3.40-3.38 (d, 2H), 3.17 (s, 3H), 3.12-2.94 (m, 9H), 1.92-1.80 (m,
4H).
Example 29
N-[(20S)-6-chloro-24-methoxy-20-(morpholin-4-yl)-2,4,8,27-tetraazapentacyc-
lo[14.8.1.1.sup.3,7.1.sup.9,13.0.sup.17,23]heptacosa-1(25),3(27),4,6,9(26)-
,10,12,16,23-nonaen-10-yl]N-methylmethanesulfonamide
##STR00033##
[0246] Example 29a
N-{2-[2-(4-Bromo-1-methoxy-7-morpholin-4-yl-6,7,8,9-tetrahydro-5H-benzocyc-
lohepten-2-ylamino)-5-chloro-pyrimidin-4-ylamino]-4-vinyl-phenyl}-N-methyl-
sulfonamide
[0247]
N-{2-[2-(4-Bromo-1-methoxy-7-morpholin-4-yl-6,7,8,9-tetrahydro-5H-b-
enzocyclohepten-2-ylamino)-5-chloro-pyrimidin-4-ylamino]-4-vinyl-phenyl}-N-
-methylsulfonamide was prepared in a similar manner as
N-(2-{2-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamino]-5-chl-
oro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
(prepared as in Example 4i) after substituting
4-Bromo-1-methoxy-7-morpholin-4-yl-6,7,8,9-tetrahydro-5H-benzocyclohepten-
-2-ylamine for
5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine. The
crude reaction mixture was concentrated under reduced pressure and
the residue purified via preparative HPLC yielding
N-{2-[2-(4-Bromo-1-methoxy-7-morpholin-4-yl-6,7,8,9-tetrahydro-5H-benzocy-
clohepten-2-ylamino)-5-chloro-pyrimidin-4-ylamino]-4-vinyl-phenyl}-N-methy-
lsulfonamide as a TFA salt white lyophylate in 30% yield, which was
used for the subsequent reaction without further manipulation. LC:
94%; LC/MS: M+H=693.0.
Example 29b
N-[(14Z,20S)-6-chloro-24-methoxy-20-(morpholin-4-yl)-2,4,8,27-tetraazapent-
acyclo[14.8.1.1.sup.3,7.1.sup.9,13.1.sup.17,23]heptacosa-1(25),3(27),4,6,9-
(26),10,12,14,16,23-decaen-10-yl]N-methylmethanesulfonamide
[0248]
N-[(14Z,20S)-6-chloro-24-methoxy-20-(morpholin-4-yl)-2,4,8,27-tetra-
azapentacyclo[14.8.1.1.sup.3,7.1.sup.9,13.0.sup.17,23]heptacosa-1(25),3(27-
),4,6,9(26),10,12,14,16,23-decaen-10-yl]N-methylmethanesulfonamide
was prepared in a similar manner as Methyl
(14Z)-6-chloro-17-(4-methylpiperazin-1-yl)-2,4,8,22-tetraazatetracyclo[14-
.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-deca-
ene-10-carboxylate (prepared as in Example 16c) after substituting
N-{2-[2-(4-Bromo-1-methoxy-7-morpholin-4-yl-6,7,8,9-tetrahydro-5H-benzocy-
clohepten-2-ylamino)-5-chloro-pyrimidin-4-ylamino]-4-vinyl-phenyl}-N-methy-
lsulfonamide for
4-Bromo-2-(5-chloro-2-[4-(4-methyl-piperazin-1-yl)-3-vinyl-phenylamino]-p-
yrimidin-4-ylamino)-benzoic acid methyl ester. The desired product
N-[(14Z,20S)-6-chloro-24-methoxy-20-(morpholin-4-yl)-2,4,8,27-tetraazapen-
tacyclo[14.8.1.1.sup.3,7.1.sup.9,13.0.sup.17,23]heptacosa-1(25),3(27),4,6,-
9(26),10,12,14,16,23-decaen-10-yl]N-methylmethanesulfonamide was
isolated in 68% yield as a black tinted solid. LC: 96%; LC/MS:
M+H=611.1.
Example 29c
N-[(20S)-6-chloro-24-methoxy-20-(morpholin-4-yl)-2,4,8,27-tetraazapentacyc-
lo[14.8.1.1.sup.3,7.1.sup.9,13.0.sup.17,23]heptacosa-1(25),3(27),4,6,9(26)-
,10,12,16,23-nonaen-10-yl]N-methylmethanesulfonamide
[0249]
N-[(20S)-6-chloro-24-methoxy-20-(morpholin-4-yl)-2,4,8,27-tetraazap-
entacyclo[14.8.1.1.sup.3,7.1.sup.9,13.0.sup.17,23]heptacosa-1(25),3(27),4,-
6,9(26),10,12,16,23-nonaen-10-yl]N-methylmethanesulfonamide was
prepared in a similar manner as
6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,22-tet-
raazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),1-
0,12,16,18-nonaene (prepared as in Example 12) after substituting
N-[(14Z,20S)-6-chloro-24-methoxy-20-(morpholin-4-yl)-2,4,8,27-tetraazapen-
tacyclo[14.8.1.1.sup.3,7.1.sup.9,13.0.sup.17,23]heptacosa-1(25),3(27),4,6,-
9(26),10,12,14,16,23-decaen-10-yl]N-methylmethanesulfonamide for
(14Z)-6-Chloro-10-(propane-2-sulfonyl)-17-(4-methylpiperazin-1-yl)-2,4,8,-
22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa-1(20),3(22),4,6,9-
(21),10,12,14,16,18-decaene. The workup varied slightly in that
after the addition of water no solid precipitated, so the aqueous
solution was partitioned between saturated aqueous NaHCO.sub.3 and
EtOAc. The organic phase was dried over Na.sub.2SO.sub.4, filtered,
and concentrated under reduced pressure. The residue was purified
via preparative HPLC. The desired product
N-[(20S)-6-chloro-24-methoxy-20-(morpholin-4-yl)-2,4,8,27-tetraazapentacy-
clo[14.8.1.1.sup.3,7.1.sup.9,13.0.sup.17,23]heptacosa-1(25),3(27),4,6,9(26-
),10,12,16,23-nonaen-10-yl]N-methylmethanesulfonamide was isolated
as a TFA salt, white lyophylate in 41% yield. LC: 97%; LC/MS:
M+H=613.1; .sup.1H NMR (DMSO-d6) .delta. 9.51 (bm, 1H,
exchangeable), 8.62 (s, 1H, exchangeable), 8.27 (s, 1H,
exchangeable), 8.24 (s, 1H), 8.21 (s, 1H), 7.58 (s, 1H), 7.48-7.46
(d, 1H), 7.06-7.04 (d, 1H), 3.98-3.95 (d, 2H), 3.72-3.66 (m, 2H),
3.59 (s, 3H), 3.50-2.94 (bm, 17H+water), 2.58-2.37 (4H+DMSO),
1.44-1.39 (m, 2H).
Example 30
6-Trifluoromethyl-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]-
docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene Trifluoroacetate
(1:1)
##STR00034##
[0250] Example 30a
(3-Bromo-phenyl)-2-chloro-5-trifluoromethyl-pyrimidin-4-yl)-amine
[0251]
(3-Bromo-phenyl)-2-chloro-5-trifluoromethyl-pyrimidin-4-yl)-amine
was prepared in a similar manner as
(2,5-Dichloro-pyrimidin-4-yl)-(2-methoxy-5-vinyl-phenyl)-amine
(prepared as in Example 13c) after substituting
2,4-dichloro-5-trifluoromethylpyrimidine for
2,4,5-trichloro-pyrimidine and 3-bromoaniline for
2-methoxy-5-vinyl-phenylamine. The crude mixture was concentrated
under reduced pressure and partitioned between organic and
saturated aqueous NaHCO.sub.3. The organic phase was dried under
Na.sub.2SO.sub.4, filtered, and concentrated. The crude
(3-Bromo-phenyl)-2-chloro-5-trifluoromethyl-pyrimidin-4-yl)-amine
was purified by normal phase chromatography (ethyl acetate/hexane),
after which it was used for the subsequent reaction without further
manipulation.
Example 30b
N(4)-(3-Bromo-phenyl)-5-trifluoromethyl-N(2)-(3-vinyl-phenyl)-pyrimidine-2-
,4-diamine
[0252]
N(4)-(3-Bromo-phenyl)-5-trifluoromethyl-N(2)-(3-vinyl-phenyl)-pyrim-
idine-2,4-diamine was prepared in a similar manner as
N-(2-{2-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamino]-5-chl-
oro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide
(prepared as in Example 4i) after substituting
(3-Bromo-phenyl)-2-chloro-5-trifluoromethyl-pyrimidin-4-yl)-amine
for
N-[2-(2,5-Dichloro-pyrimidin-4-ylamino)-4-vinyl-phenyl]-N-methyl-methanes-
ulfonamide, 3-vinyl-phenylamine for
5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamine, and 4M
HCl in dioxane for methanesulfonic acid. The crude
N(4)-(3-Bromo-phenyl)-5-trifluoromethyl-N(2)-(3-vinyl-phenyl)-pyrimidine--
2,4-diamine was purified via preparative reverse phase HPLC, then
free based by partitioning between CHCl.sub.3 and saturated aqueous
NaHCO.sub.3. The organic phase was dried over Na.sub.2SO.sub.4,
filtered, and concentrated to give
N(4)-(3-Bromo-phenyl)-5-trifluoromethyl-N(2)-(3-vinyl-phenyl)-pyrimidine--
2,4-diamine, which was used for the subsequent step without further
manipulation.
Example 30c
(14Z)-6-Trifluoromethyl-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup-
.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene
[0253]
(14Z)-6-Trifluoromethyl-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,-
7.1.sup.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene
was prepared in a similar manner as
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene (prepared as in
Example 4j) after substituting
N(4)-(3-Bromo-phenyl)-5-trifluoromethyl-N(2)-(3-vinyl-phenyl)-pyrimidine--
2,4-diamine for
N-(2-{2-[5-Bromo-2-methoxy-4-(4-methyl-piperazin-1-yl)-phenylamino]-5-chl-
oro-pyrimidin-4-ylamino}-4-vinyl-phenyl)-N-methyl-methanesulfonamide.
The workup varied in that
(14Z)-6-Trifluoromethyl-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.su-
p.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene was
purified chromatographically rather than via crystallization. MP:
167-169.degree. C., LC: 95%; LC/MS: M+H=355.1.
Example 30d
6-Trifluoromethyl-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]-
docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene Trifluoroacetate
(1:1)
[0254]
6-Trifluoromethyl-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.su-
p.9,13]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene was
prepared in a similar manner as
6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin-1-yl)--
19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]docosa--
1(20),3(22),4,6,9(21),10,12,16,18-nonaene (prepared as in Example
5) after substituting
(14Z)-6-Trifluoromethyl-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.su-
p.9,13]docosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene for
(14Z)-6-Chloro-10-(2-methanesulfonyl-methyl-amino)-17-(4-methylpiperazin--
1-yl)-19-methoxy-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13]d-
ocosa-1(20),3(22),4,6,9(21),10,12,14,16,18-decaene. The desired
6-Trifluoromethyl-2,4,8,22-tetraazatetracyclo[14.3.1.1.sup.3,7.1.sup.9,13-
]docosa-1(20),3(22),4,6,9(21),10,12,16,18-nonaene was isolated as a
white lyophylate. LC: 100%, LC/MS: M+H=357.1.
VI. Biology
[0255] ALK Kinase Assay
[0256] Example compounds were tested for their ability to inhibit
the kinase activity of baculovirus-expressed ALK using a
modification of the ELISA protocol reported for trkA in Angeles, T.
S. et al., Anal. Biochem. 1996, 236, 49-55, which is incorporated
herein by reference in its entirety. Phosphorylation of the
substrate, phospholipase C-gamma (PLC-.gamma.) generated as a
fusion protein with glutathione S-transferase (GST) as reported in
Rotin, D. et al., EMBO J. 1992, 11, 559-567, which is incorporated
herein by reference in its entirety, was detected with a
europium-labeled anti-phosphotyrosine antibody and measured by
time-resolved fluorescence (TRF). Briefly, each 96-well plate was
coated with 100 .mu.L/well of 10 .mu.g/mL substrate
(GST-PLC-.gamma.) in Tris-buffered saline (TBS). The assay mixture
(total volume=100 .mu.L/well) consisting of 20 mM HEPES, pH 7.2, 1
.mu.M ATP (K.sub.m level), 5 mM MnCl.sub.2, 0.1% BSA, 2.5% DMSO,
and various concentrations of test compound was then added to the
assay plate. The reaction was initiated by adding enzyme (30 ng/mL
ALK) and was allowed to proceed at 37.degree. C. for 15 minutes.
Detection of the phosphorylated product was performed by adding 100
.mu.L/well of Eu-N1 labeled PT66 antibody (Perkin Elmer # AD0041).
Incubation at 37.degree. C. then proceeded for 1 hour, followed by
addition of 100 .mu.L enhancement solution (Wallac #1244-105). The
plate was gently agitated and after thirty minutes, the
fluorescence of the resulting solution was measured using the
PerkinElmer EnVision.TM. 2102 (or 2104) multilabel plate
reader.
[0257] Data analysis was performed using ActivityBase (IDBS,
Guilford, UK). IC.sub.50 values were calculated by plotting percent
inhibition versus log.sub.10 of the concentration of compound and
fitting to the nonlinear regression sigmoidal dose-response
(variable slope) equation in XLFit (IDBS, Guilford, UK).
[0258] JAK2 Kinase Assay
[0259] Compounds were tested for their ability to inhibit the
kinase activity of baculovirus-expressed JAK2 using the TRF
detection system. IC.sub.50 runs were conducted in 96-well Costar
high binding plates (Corning Costar #3922, Corning, N.Y.). The
plates were coated first with 100 .mu.L/well of 10 g/mL Neutravidin
(Pierce #31000, Rockford, Ill.) in TBS at 37.degree. C. for 2
hours, followed by 100 .mu.L/well of 1 g/mL 15-mer peptide
substrate (biotinyl-amino-hexanoyl-EQEDEPEGDYFEWLE-amide, Infinity
Biotech Research and Resource, Aston, Pa.) at 37.degree. C. for
another hour. The JAK2 assay mixture (total volume=100 .mu.L/well)
consisting of 20 mM HEPES, pH 7.2, 0.2 M ATP, 1 mM MnCl.sub.2, 0.1%
BSA, and test compound (diluted in DMSO; 2.5% DMSO final in assay)
was then added to the assay plate. Enzyme (15 ng/ml JAK2) was added
and the reaction was allowed to proceed at room temperature for 20
minutes. Detection of the phosphorylated product was performed by
adding 100 .mu.L/well of Eu-N1 labeled PY100 antibody (PerkinElmer
Life Sciences #AD0041, Boston, Mass.). Incubation at the room
temperature then proceeded for 1 hour, followed by addition of 100
.mu.l enhancement solution (PerkinElmer Life Sciences #1244-105,
Boston, Mass.). The plate was gently agitated and after thirty
minutes, the fluorescence of the resulting solution was measured
using the PerkinElmer EnVision.TM. 2102 (or 2104) multi-label plate
reader.
[0260] Data analysis was performed using ActivityBase (IDBS,
Guilford, UK). IC.sub.50 values were calculated by plotting percent
inhibition versus log.sub.10 of the concentration of compound and
fitting to the nonlinear regression sigmoidal dose-response
(variable slope) equation in XLFit (IDBS, Guilford, UK).
[0261] FAK Enzyme Assay
[0262] Example compounds were tested for their ability to inhibit
the kinase activity of baculovirus-expressed recombinant human FAK
using the TRF detection system as described above for JAK2.
IC.sub.50 runs were performed in 96-well Costar high binding plates
(#3922). The plates were coated first with 100 .mu.L/well of 10
.mu.g/mL neutravidin in TBS at 37.degree. C. for 2 h, followed by
100 .mu.L/well of 1 g/mL 15-mer peptide substrate
(biotinyl-amino-hexanoyl-EQEDEPEGDYFEWLE-amide) at 37.degree. C.
for another hour. The FAK assay mixture (total volume=100
.mu.L/well) consisting of 20 mM HEPES, pH 7.2, 10 .mu.M ATP, 5 mM
MgCl.sub.2, 0.5 mM DTT, 0.1% BSA, and test compound (diluted in
DMSO; 2.5% DMSO final in assay) was then added to the assay plate.
Enzyme (10 ng/mL FAK, Invitrogen #PV3832) was added and the
reaction was allowed to proceed at room temperature for 30 minutes.
Detection of the phosphorylated product was performed by adding 100
.mu.L/well of Eu-N1 labeled PY100 antibody (diluted 1:75000 in
antibody dilution buffer). Samples were incubated at room
temperature for 1 hour, followed by addition of 100 .mu.L
enhancement solution. Plates were agitated for 10 minutes and
fluorescence of the resulting solution measured using the
PerkinElmer EnVision.TM. 2102 (or 2104) multi-label plate
reader.
[0263] Data analysis was performed using ActivityBase (IDBS,
Guilford, UK). IC.sub.50 values were calculated by plotting percent
inhibition versus log.sub.10 of the concentration of compound and
fitting to the nonlinear regression sigmoidal dose-response
(variable slope) equation in XLFit (IDBS, Guilford, UK).
[0264] Insulin Receptor Kinase Assay
[0265] Example compounds were tested for their ability to inhibit
the kinase activity of baculovirus-expressed human insulin receptor
cytoplasmic domain (.beta.IR.sub.CD) using the TRF assay as
described above for ALK. Phosphorylation of the substrate,
recombinant GST-PLC-.gamma., was detected with a europium-labeled
anti-phosphotyrosine antibody and measured by TRF. Each 96-well
plate (Greiner#655074) was coated with 100 .mu.L/well of 20
.mu.g/mL substrate solution in TBS. The IR assay mixture (total
volume=100 .mu.L/well) consisting of 20 mM HEPES (pH 7.2), 20 .mu.M
ATP, 5 mM MnCl.sub.2, 0.1% BSA, and test compound (diluted in DMSO;
2.5% DMSO final in assay) was added to the assay plates. Enzyme (20
ng/mL .beta.IR.sub.CD) was added and the reaction was allowed to
proceed at room temperature for 20 minutes. Detection of the
phosphorylated product was performed by adding 100 .mu.L/well of
Eu-N1 labeled PY100 antibody (PerkinElmer# AD0160; diluted 1:10,000
in TBS-T containing 0.25% BSA). Incubation at 37.degree. C. for 1
hour was followed by addition of 50 .mu.L enhancement solution
(Wallace#1244-105). Plates were agitated for 10 minutes and the
fluorescence was measured using the PerkinElmer EnVision.TM. 2102
or 2104 multi-label plate reader.
[0266] Data analysis was performed using ActivityBase (IDBS,
Guilford, UK). IC.sub.50 values were calculated by plotting percent
inhibition versus log.sub.10 of the concentration of compound and
fitting to the nonlinear regression sigmoidal dose-response
(variable slope) equation in XLFit (IDBS, Guilford, UK).
[0267] Results
[0268] Biological data for Example compounds is presented in the
following Table 1. Unless otherwise specified in Table 1, IC.sub.50
nanomolar value ranges designated as A, B, C, D or E, indicate the
following ranges:
[0269] IC.sub.50<10 nM A;
[0270] IC.sub.50 10 nM to 100 nM B;
[0271] IC.sub.50 101 nM to 1,000 nM C;
[0272] IC.sub.50 1,001 nM to 10,000 nM D; and
[0273] IC.sub.50>10,000 nM E.
[0274] "NT" denotes not tested.
[0275] Unless otherwise specified, all values are an average of two
or more determinations.
TABLE-US-00001 TABLE 1 ALK, JAK2, FAK, Insulin Receptor Kinase
Inhibition Example FAK IC.sub.50 ALK IC.sub.50 JAK2 IC.sub.50 IR
IC.sub.50 No. (nM) (nM) (nM) (nM) 1 E E C NT 24 E >3000 A E 25 E
E A E 2 C C A D 3 C B A C 4 >3000 C A D 26 >1000 D A >3000
21 E E A E 20 NT NT NT NT 14 B A A C 13 C C B >3000 15 C A B C
22 C C A D 23 B C A C 30 D >3000 C NT 18 B C B NT 16 C C B NT 27
D C B NT 19 B B B C 17 A B A C 11 B B A D 12 A A A B 9 A B A
>2000 10 A A A C 29 B B C >3000 4 A A C >2000 5 A A B B 6
A A C B 7 A B D >2000 8 A A C C 28 A A B C
* * * * *