U.S. patent application number 14/295398 was filed with the patent office on 2014-09-25 for heteroaryl compounds and their uses.
This patent application is currently assigned to NOVARTIS AG. The applicant listed for this patent is Paul A. Barsanti, Cheng Hu, Jeff Jin, Robert Keyes, Robert Kucejko, Xiaodong Lin, Yue Pan, Keith B. Pfister, Martin Sendzik, James Sutton, Lifeng Wan. Invention is credited to Paul A. Barsanti, Cheng Hu, Jeff Jin, Robert Keyes, Robert Kucejko, Xiaodong Lin, Yue Pan, Keith B. Pfister, Martin Sendzik, James Sutton, Lifeng Wan.
Application Number | 20140288076 14/295398 |
Document ID | / |
Family ID | 42727614 |
Filed Date | 2014-09-25 |
United States Patent
Application |
20140288076 |
Kind Code |
A1 |
Barsanti; Paul A. ; et
al. |
September 25, 2014 |
HETEROARYL COMPOUNDS AND THEIR USES
Abstract
The present invention provides a compound of formula (I):
##STR00001## and pharmaceutically acceptable salts, enantiomers,
stereoisomers, rotamers, tautomers, diastereomers, or racemates
thereof. Also provided is a method of treating a disease or
condition mediated by CDK9.
Inventors: |
Barsanti; Paul A.; (Pleasant
Hill, CA) ; Hu; Cheng; (Menlo Park, CA) ; Jin;
Jeff; (San Ramon, CA) ; Keyes; Robert;
(Pleasant Paraie, WI) ; Kucejko; Robert;
(Philadelphia, PA) ; Lin; Xiaodong; (Walnut Creek,
CA) ; Pan; Yue; (Albany, CA) ; Pfister; Keith
B.; (San Ramon, CA) ; Sendzik; Martin; (San
Mateo, CA) ; Sutton; James; (Pleasanton, CA) ;
Wan; Lifeng; (Richmond, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Barsanti; Paul A.
Hu; Cheng
Jin; Jeff
Keyes; Robert
Kucejko; Robert
Lin; Xiaodong
Pan; Yue
Pfister; Keith B.
Sendzik; Martin
Sutton; James
Wan; Lifeng |
Pleasant Hill
Menlo Park
San Ramon
Pleasant Paraie
Philadelphia
Walnut Creek
Albany
San Ramon
San Mateo
Pleasanton
Richmond |
CA
CA
CA
WI
PA
CA
CA
CA
CA
CA
CA |
US
US
US
US
US
US
US
US
US
US
US |
|
|
Assignee: |
NOVARTIS AG
Basel
CH
|
Family ID: |
42727614 |
Appl. No.: |
14/295398 |
Filed: |
June 4, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13773052 |
Feb 21, 2013 |
8778951 |
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14295398 |
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12843494 |
Jul 26, 2010 |
8415381 |
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13773052 |
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Current U.S.
Class: |
514/237.2 ;
514/235.5; 514/253.01; 514/255.05; 514/256; 514/275; 514/278;
514/318; 514/333; 514/334 |
Current CPC
Class: |
C07D 401/04 20130101;
A61P 31/18 20180101; C07D 413/14 20130101; A61P 33/00 20180101;
C07D 405/14 20130101; A61P 13/10 20180101; A61P 25/28 20180101;
A61P 43/00 20180101; A61P 37/06 20180101; A61P 1/18 20180101; A61P
9/00 20180101; A61P 35/00 20180101; C07D 241/20 20130101; A61P
13/12 20180101; A61P 17/06 20180101; C07D 401/12 20130101; A61P
31/12 20180101; A61P 3/10 20180101; A61P 35/02 20180101; A61K
31/497 20130101; A61P 1/00 20180101; A61P 9/10 20180101; A61P 31/10
20180101; A61P 19/00 20180101; A61P 19/02 20180101; C07D 409/14
20130101; A61P 11/00 20180101; A61P 15/00 20180101; A61P 29/00
20180101; A61P 37/02 20180101; C07D 213/74 20130101; C07D 401/14
20130101 |
Class at
Publication: |
514/237.2 ;
514/334; 514/275; 514/318; 514/255.05; 514/333; 514/256; 514/278;
514/253.01; 514/235.5 |
International
Class: |
C07D 409/14 20060101
C07D409/14; C07D 401/14 20060101 C07D401/14; C07D 405/14 20060101
C07D405/14; C07D 401/04 20060101 C07D401/04 |
Claims
1. A method to treat a disease or condition selected from cancer,
cardiac hypertrophy, HIV and inflammatory diseases, which comprises
administering to a subject in need thereof a therapeutically
effective amount of a compound of the Formula: ##STR00507## or a
pharmaceutically acceptable salt thereof, wherein: A.sub.1 is
CR.sub.6; A.sub.2 is CR.sub.7; A.sub.3 is CR.sub.8; A.sub.4 is
NR.sub.9 or O; L is selected from optionally substituted
C.sub.1-4alkyl, C.sub.3-6 cycloalkyl, C.sub.3-6 heterocycloalkyl,
or C.sub.2-4 alkenyl; R.sub.1 is X--R.sub.16; X is a bond or
C.sub.1-4 alkyl; R.sub.16 is selected from the group consisting of
C.sub.1-6alkyl, C.sub.3-6branched alkyl, C.sub.3-8cycloalkyl,
heterocycloalkyl, C.sub.3-8-partially unsaturated cycloalkyl, aryl,
and heteroaryl; wherein R.sub.16 is substituted with one to three
groups independently selected from halogen, hydrogen,
C.sub.1-6alkyl, C.sub.1-6haloalkyl, C.sub.3-6branched alkyl,
C.sub.3-4branched haloalkyl, OH, C.sub.1-6alkoxy,
R.sub.22--OR.sub.12, S(O).sub.0-2R.sub.12,
R.sub.22--S(O).sub.0-2R.sub.12, S(O).sub.2NR.sub.13R.sub.14,
R.sub.22--S(O).sub.2NR.sub.13R.sub.14, C(O)OR.sub.12,
R.sub.22--C(O)OR.sub.12, C(O)R.sub.19, R.sub.22--C(O)R.sub.19,
O--C.sub.1-3 alkyl, OC.sub.1-3 haloalkyl, OC(O)R.sub.19,
R.sub.22--OC(O)R.sub.19, C(O)NR.sub.13R.sub.14,
R.sub.22--C(O)NR.sub.13R.sub.14, NR.sub.15S(O).sub.2R.sub.12,
R.sub.22--NR.sub.15S(O).sub.2R.sub.12, NR.sub.17R.sub.18,
R.sub.22--NR.sub.17R.sub.18, NR.sub.15C(O)R.sub.19,
R.sub.22--NR.sub.15C(O)R.sub.19, NR.sub.15C(O)OCH.sub.2Ph,
R.sub.22--NR.sub.15C(O)OCH.sub.2Ph, NR.sub.15C(O)OR.sub.12,
R.sub.22--NR.sub.15C(O)OR.sub.12, NR.sub.15C(O)NR.sub.13R.sub.14,
and R.sub.22--NR.sub.15C(O)NR.sub.13R.sub.14; R.sub.17 and R.sub.18
are each, independently, selected from the group consisting of
hydrogen, hydroxyl, C.sub.1-6alkyl, C.sub.1-6haloalkyl,
C.sub.3-6branched alkyl, C.sub.3-6 cycloalkyl, R.sub.22--OR.sub.12,
R.sub.22--S(O).sub.0-2R.sub.12,
R.sub.22--S(O).sub.2NR.sub.13R.sub.14, R.sub.22--C(O)OR.sub.12,
R.sub.22--C(O)R.sub.19, R.sub.22--OC(O)R.sub.19,
R.sub.22--C(O)NR.sub.13R.sub.14,
R.sub.22--NR.sub.15S(O).sub.2R.sub.12, R.sub.22--NR.sub.23R.sub.24,
R.sub.22--NR.sub.15C(O)R.sub.19,
R.sub.22--NR.sub.15C(O)OCH.sub.2Ph,
R.sub.22--NR.sub.15C(O)OR.sub.12,
R.sub.22--NR.sub.15C(O)NR.sub.13R.sub.14, cycloalkyl,
heterocycloalkyl and heteroaryl; alternatively, R.sub.17 and
R.sub.18 along with the nitrogen atom to which they are attached to
can be taken together to form a four to six membered heterocyclic
ring wherein the carbon atoms of said ring are optionally
substituted with R.sub.20, and the nitrogen atoms of said ring are
optionally substituted with R.sub.21; R.sub.19 is selected from
optionally substituted alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, and optionally substituted heteroaryl; R.sub.20 is selected
from the group consisting of C.sub.1-6 alkyl or C.sub.1-6
haloalkyl; R.sub.21 is selected from the group consisting of
C.sub.1-6alkyl, C.sub.1-6haloalkyl, C(O)R.sub.12, C(O)OR.sub.12,
S(O).sub.2R.sub.12; R.sub.22 is selected from the group consisting
of C.sub.1-6 alkyl, C.sub.1-6 haloalkyl, C.sub.3-6 branched alkyl,
C.sub.3-6branched haloalkyl; R.sub.23 and R.sub.24 are each,
independently, selected from the group consisting of hydrogen,
C.sub.1-6 alkyl, C.sub.1-6haloalkyl, C.sub.3-6 branched alkyl,
C.sub.3-6 branched haloalkyl; R.sub.2 is selected from the group
consisting of optionally substituted C.sub.1-6 alkyl, optionally
substituted C.sub.3-6 branched alkyl, optionally substituted
C.sub.3-6 cycloalkyl, optionally substituted heterocycloalkyl,
optionally substituted aryl, and optionally substituted heteroaryl;
R.sub.4, R.sub.5, and R.sub.6 are each, independently, selected
from the group consisting of hydrogen, hydroxyl, cyano, halogen,
C.sub.1-4 alkyl, C.sub.1-4haloalkyl, C.sub.2-4 alkenyl, C.sub.2-4
alkynyl, amino, NR.sub.10R.sub.11, and alkoxy; R.sub.3 and R.sub.7
are each, independently, selected from the group consisting of
hydrogen, hydroxyl, cyano, halogen, alkyl, haloalkyl, alkenyl,
alkynyl, alkoxy, NR.sub.10R.sub.11, C(O)R.sub.12, C(O)OR.sub.12,
C(O)NR.sub.13R.sub.14, S(O).sub.0-2R.sub.12,
S(O).sub.0-2NR.sub.13R.sub.14, and optionally substituted C.sub.3-4
cycloalkyl; R.sub.8 is selected from Cl, F, and methyl; R.sub.9 is
selected from the group consisting of hydrogen, C.sub.1-4 alkyl,
alkoxy, C(O)R.sub.12, C(O)OR.sub.15, C(O)NR.sub.13R.sub.14,
S(O).sub.0-2R.sub.12, S(O).sub.0-2NR.sub.13R.sub.14, optionally
substituted C.sub.3-4 cycloalkyl, and optionally substituted
heterocycloalkyl; R.sub.10 and R.sub.11 are each, independently,
selected from the group consisting of hydrogen, hydroxyl, alkyl,
alkoxy, C(O)R.sub.12, C(O)OR.sub.12, C(O)NR.sub.13R.sub.14,
S(O).sub.0-2R.sub.12, and S(O).sub.0-2NR.sub.13R.sub.14;
alternatively, R.sub.10 and R.sub.11 along with the nitrogen atom
to which they are attached to can be taken together to form an
optionally substituted four to six membered heteroaromatic, or a
non-aromatic heterocyclic ring; R.sub.12 and R.sub.15 are each,
individually, selected from the group consisting of hydrogen,
alkyl, branched alkyl, haloalkyl, branched haloalkyl,
(CH.sub.2).sub.0-3-cycloalkyl, (CH.sub.2).sub.0-3-heterocycloalkyl,
(CH.sub.2).sub.0-3-aryl, and heteroaryl; R.sub.13 and R.sub.14 are
each, independently, selected from the group consisting of
hydrogen, hydroxyl, alkyl, branched alkyl, haloalkyl, branched
haloalkyl, alkoxy, cycloalkyl or heterocycloalkyl; and
alternatively, R.sub.13 and R.sub.14 along with the nitrogen atom
to which they are attached to can be taken together to form an
optionally substituted four to six membered heteroaromatic, or
non-aromatic heterocyclic ring.
2. The method of claim 1, wherein: R.sub.8 is Cl.
3. The method of claim 1, wherein: R.sub.1 is X--R.sub.16; X is a
bond, or C.sub.1-2 alkyl; R.sub.16 is selected from the group
consisting of C.sub.1-2-alkyl, C.sub.4-6cycloalkyl,
heterocycloalkyl, phenyl, and heteroaryl; wherein R.sub.16 is
substituted with one to three groups independently selected from
halogen, hydrogen, C.sub.1-3alkyl, C.sub.3-6branched alkyl, OH,
C.sub.1-2alkoxy, R.sub.22--OR.sub.12, S(O).sub.1-2R.sub.12,
C(O)OR.sub.12, R.sub.22--C(O)OR.sub.12, C(O)R.sub.19,
R.sub.22--OC(O)R.sub.19, C(O)NR.sub.13R.sub.14,
NR.sub.15S(O).sub.2R.sub.12, NR.sub.17R.sub.18,
R.sub.22--NR.sub.17R.sub.18, NR.sub.15C(O)R.sub.19,
R.sub.22--NR.sub.15C(O)R.sub.19, and NR.sub.15C(O)OCH.sub.2Ph.
4. The method of claim 1, wherein: R.sub.16 is selected from the
group consisting of C.sub.1-2-alkyl, cyclopentyl, cyclohexyl,
piperidine, piperazine, morpholine, pyridine, pyrrolidine,
cyclohexenyl, and tetrahydro-2H-pyran; wherein R.sub.16 is
substituted with one to three groups selected from amino, hydroxyl,
NHCH.sub.2-phenyl, CH.sub.2-amino, COO-t-butyl, H, methoxy,
NH--SO.sub.2-ethyl, CH.sub.2--NHSO.sub.2-ethyl, SO.sub.2-ethyl,
t-butyl, methyl, CH.sub.2--COOH, CO--NHCH.sub.3,
CON(CH.sub.3).sub.2, NHC(CH.sub.3)--CH.sub.2--SO.sub.2--CH.sub.3,
NH--COO--CH.sub.2-phenyl, hydroxy-methyl, CH.sub.2--NH--CH.sub.3,
CH.sub.2--NH-ethyl, NH--CH.sub.2--CH.sub.2-methoxy,
CH.sub.2--NH--CO--CH.sub.3, NH--CH.sub.2--CH.sub.2OH,
NH--CO--CH.sub.2--N(CH.sub.3).sub.2, NH--CO-methylpyrrolidine,
NH--CH.sub.2--C(CH.sub.3)-dioxolane, NH--CO-pyridyl, NH-ethyl,
pyrrolidine, CH.sub.2--NH--CO-pyridyl, NH-tetrahydropyran,
COCH.sub.2--N(CH.sub.3).sub.2,
NH--CH.sub.2--C(CH.sub.3)-dimethyldioxolane, tetrahydropyran,
CO-methylpyrrolidine, CH.sub.2-methylpiperidine, NH--CO--CH.sub.3,
NH--SO.sub.2--CH.sub.3, NH--CH(CH.sub.2--OCH.sub.3).sub.2,
NH--CH.sub.2-tetrahydrofuran, NH--CH.sub.2-oxetane,
NH-tetrahydropyran, NH--CH.sub.2-dioxane,
N(CH.sub.3)--CH.sub.2CH.sub.2--OCH.sub.3, CH(OH)--CH.sub.2-amino,
NH--CH.sub.2CH.sub.2--OCF.sub.3, NHCH.sub.2--OCH.sub.3,
NH--CH.sub.2--CH(CF.sub.3)--OCH.sub.3,
NH--CH(CH.sub.3)--CH.sub.2--OH, F, NH-oxetane,
CH.sub.2--CH.sub.2--OCH.sub.3, CH.sub.2--OCH.sub.3,
CH.sub.2-tetrahydropyran, CH.sub.2-methylpiperizine,
NH.sub.2--CH.sub.2--CH(OH)--CF.sub.3, piperidine,
CH.sub.2-pyrrolidine, NH--CH(CH.sub.3)CH.sub.2OCH.sub.3,
NH-tetrahydrofuran, (CH.sub.2).sub.3--NH.sub.2, hydroxyethyl,
propyl, CH.sub.2-pyridyl, CH.sub.2-piperidine, morpholine,
NH-chloropyrimidine, NH--CH.sub.2CH.sub.2--SO.sub.2-methyl,
(CH.sub.3).sub.3--N(CH.sub.3).sub.2, piperizine, ##STR00508## and
CH.sub.2-morpholine.
5. The method of claim 1 wherein: R.sub.3 is selected from H,
methyl, cyano, chloro, CONH.sub.2, amino, cyclopropyl, ethyl, and
fluoro; R.sub.4 is selected from halogen, methyl, hydrogen, and
halo-methyl; R.sub.6 is H; R.sub.7 is selected from H, COOH, Cl, F,
CONH.sub.2, CN, and CF.sub.3; R.sub.8 is Cl; R.sub.17 and R.sub.18
are each, independently, selected from the group consisting of
hydrogen, C.sub.1-3alkyl, C.sub.1-4haloalkyl, C.sub.3-6branched
alkyl, R.sub.22--OR.sub.12, R.sub.22--S(O).sub.2R.sub.12,
R.sub.22--NR.sub.15S(O).sub.2R.sub.12, heterocycloalkyl or
heteroaryl; alternatively, R.sub.17 and R.sub.18 along with the
nitrogen atom to which they are attached to can be taken together
to form a four to six membered heterocyclic ring wherein said ring
carbon atoms are optionally substituted with R.sub.20, and the ring
nitrogen atoms are optionally substituted with R.sub.21; R.sub.19
is selected from C.sub.1-3-alkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl or optionally
substituted heteroaryl; R.sub.20 represents the group
C.sub.1-3alkyl; and R.sub.22 is selection from the group consisting
of C.sub.1-4alkyl, and C.sub.3-6 branched alkyl.
6. The method of claim 1, wherein: L is selected from
C.sub.1-4-alkyl, and cyclopropyl; R.sub.2 is selected from the
group consisting of C.sub.3-7 cycloalkyl, a five to seven membered
heterocycloalkyl, phenyl, and pyridyl, wherein each said R.sub.2
group is substituted with one, two, or three substituents
independently selected from hydrogen, cyano, CO--NH.sub.2, halogen,
methoxy, dihalo-methoxy, trihalo-methoxy, trihalo alkyl,
C.sub.1-3-alkyl, and hydroxy; and R.sub.9 represents methyl,
hydrogen, or ethyl.
7. The method of claim 1, wherein: L is C.sub.1-2 alkyl, or
C.sub.3-4-cycloalkyl; R.sub.1 is X--R.sub.16; X is a bond, or
C.sub.1-2 alkyl; R.sub.16 is selected from the group consisting of
C.sub.1-2-alkyl, cyclopentyl, cyclohexyl, piperidine, piperazine,
morpholine, pyridine, pyrrolidine, cyclohexenyl, and
tetrahydro-2H-pyran; wherein R.sub.16 is substituted with one to
three groups independently selected from amino, hydroxyl,
NHCH.sub.2-phenyl, CH.sub.2-amino, COO-t-butyl, H, methoxy,
NH--SO.sub.2-ethyl, CH.sub.2--NHSO.sub.2-ethyl, SO.sub.2-ethyl,
t-butyl, methyl, CH.sub.2--COOH, CO--NHCH.sub.3,
CON(CH.sub.3).sub.2, NHCH.sub.2--CH.sub.2--SO.sub.2--CH.sub.3,
NH--COO--CH.sub.2-phenyl, hydroxy-methyl, CH.sub.2--NH--CH.sub.3,
CH.sub.2--NH-ethyl, NH--CH.sub.2--CH.sub.2-methoxy,
CH.sub.2--NH--CO--CH.sub.3, NH--CH.sub.2--CH.sub.2OH,
NH--CO--CH.sub.2--N(CH.sub.3).sub.2, NH--CO-methylpyrrolidine,
NH--CO-pyridyl, NH-ethyl, pyrrolidine, CH.sub.2--NH--CO-pyridyl,
COCH.sub.2--N(CH.sub.3).sub.2, tetrahydropyran,
CO-methylpyrrolidine, CH.sub.2-methylpiperidine, NH--CO--CH.sub.3,
NH--SO.sub.2--CH.sub.3, NH--CH.sub.2-tetrahydrofuran,
NH--CH.sub.2-dioxane, N(CH.sub.3)--CH.sub.2CH.sub.2--OCH.sub.3,
CH(OH)--CH.sub.2-amino, NH--CH.sub.2CH.sub.2--OCF.sub.3,
NH(CH.sub.3)--CH.sub.2--OCH.sub.3,
NH--CH.sub.2--CH(CF.sub.3)--OCH.sub.3, F, NH-oxetane,
CH.sub.2--CH.sub.2--OCH.sub.3, CH.sub.2--OCH.sub.3,
CH.sub.2-tetrahydropyran, CH.sub.2-methylpiperizine,
NH.sub.2--CH.sub.2--CH(OH)--CF.sub.3, piperidine,
CH.sub.2-pyrrolidine, NH--CH(CH.sub.3)CH.sub.2OCH.sub.3,
NH-tetrahydrofuran, (CH.sub.2).sub.3--NH.sub.2, hydroxyethyl,
propyl, CH.sub.2-pyridyl, CH.sub.2-piperidine, morpholine,
NH-chloropyrimidine, NH--CH.sub.2CH.sub.2--SO.sub.2-methyl,
(CH.sub.3).sub.3--N(CH.sub.3).sub.2, piperizine,
CH.sub.2-morpholine, NH--CH.sub.2--C(CH.sub.3)-dioxolane,
NH-tetrahydropyran, NH--CH.sub.2--C(CH.sub.3)-dimethyldioxolane,
NH--CH(CH.sub.2--OCH.sub.3).sub.2, NH--CH.sub.2-oxetane,
NH-tetrahydropyran, N(CH.sub.3)--CH.sub.2CH.sub.2--OCH.sub.3,
NH--CH(CH.sub.3)--CH.sub.2--OH, ##STR00509## and
NH--CH(CH.sub.3)--CH.sub.2--OH; R.sub.2 is selected from the group
consisting of cyclohexyl, 1,3-dioxane, pyridinyl, phenyl,
tetrahydropyranyl, cycloheptyl, 1,4-dioxane, morpholinyl, alkyl
substituted dioxane, tetrahydrofuranyl, dioxepane, piperidinyl, and
##STR00510## wherein each said R.sub.2 group is substituted with
one, two, or three substituents independently selected from Cl, Br,
F, methoxy, hydroxy-methyl, hydrogen, carboxamide, cyano,
dihalo-methoxy, trihalo-methoxy, trifluoro-methyl, hydroxyl, and
methyl; and R.sub.4, is chloro, hydrogen, trifluoro-methyl, fluoro,
or bromo; R.sub.5, and R.sub.6 are each independently hydrogen;
R.sub.3 is selected from hydrogen, fluoro, cyano, CO--NH.sub.2,
chloro, amino, methyl, and cyclopropyl; R.sub.7 is selected from H,
trifluoro-methyl, COOH, CO--NH.sub.2, and cyano; R.sub.8 represents
Cl; and R.sub.9 is selected from the group consisting of H, ethyl,
and methyl.
8. The method of claim 1 wherein the compound is selected from:
N2'-(trans-4-aminocyclohexyl)-5'-chloro-3,5-difluoro-N6-((tetrahydro-2H-p-
yran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-fluoro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine;
3,5'-dichloro-N2'-(trans-4-(((R)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)am-
inocyclohexyl)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6--
diamine;
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)dideuteromethyl)-N2'-(tra-
ns-4-(((S)-tetrahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2'-
,6-diamine;
5'-chloro-5-fluoro-N2'-(trans-4-(2-(methylsulfonyl)ethylamino)cyclohexyl)-
-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-5-fluoro-N2'-(trans-4-(oxetan-2-yl-methylamino)cyclohexyl)-N6-(-
(tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-(((S)-tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2',6-diamine;
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-(((R)-tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2',6-diamine;
4-((5'-chloro-2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexylamino-
)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-(3-fluorobenzyl)-2,4'-
-bipyridine-2',6-diamine;
2'-(trans-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'--
bipyridine-5-carbonitrile;
N2'-(trans-4-aminocyclohexyl)-3-chloro-5'-fluoro-N6-((tetrahydro-2H-pyran-
-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-N6-(3-fluorobenzyl)-N2'-((1R,3S)-3-if
methylamino)methyl)cyclopentyl)-2,4'-bipyridine-2',6-diamine;
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-3-fluoro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexy-
l)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-5-fluoro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexy-
l)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-N6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)-N2-(trans-4-(-
(R)-1-methoxypropan-2-yl-amino)cyclohexyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-N6-(((S)-2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)-N2'-(tran-
s-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-2,4'-bipyridine-2',6-diami-
ne;
5'-chloro-5-fluoro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4--
((1,1-dioxotetrahydro-2H-thiopyran-4-yl)methyl)aminocyclohexyl)-2,4'-bipyr-
idine-2',6-diamine;
5'-chloro-5-fluoro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tet-
rahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3,5-difluorobenzyl)-2,4'-bipy-
ridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((5-fluoropyridin-3-yl)methyl)-
-2,4'-bipyridine-2',6-diamine;
trans-4-(5'-chloro-6-(3,5-difluorobenzylamino)-2,4'-bipyridin-2-yl-amino)-
cyclohexanol;
(R)-5'-chloro-N6-(3-fluorobenzyl)-N2'-(2-(piperidin-3-yl)ethyl)-2,4'-bipy-
ridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine;
3,5'-dichloro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tetrahyd-
ro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
2-(trans-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2,4'--
bipyridin-2'-yl-amino)cyclohexylamino)ethanol;
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((R)-te-
trahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine;
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((S)-te-
trahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2'6-diamine;
3,5'-dichloro-N2'-(trans-4-((2-methoxyethyl)(methyl)amino)cyclohexyl)-N6--
((tetrahydro-2 H-pyran-4-yl)methyl)-2,4'-bipyridine-2,6-diamine;
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((R)-tetrah-
ydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((S)-tetrah-
ydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-3-fluoro-N6-((tetrahydro-2H-pyran-
-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-3-fluoro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tet-
rahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-3-bromo-5'-chloro-N6-((tetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2'6-diamine;
3-bromo-5'-chloro-N2'-(trans-4-(2-methoxyeth
ylamino)cyclohexyl)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-
-2',6-diamine; trans-4-(3,5'-dichloro-6-((2,2-di
methyltetrahydro-2H-pyran-4-yl)methyl)amino-2,4'-bipyridin-2'-yl-amino)cy-
clohexanol;
(2S)-3-(trans-4-(35'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)me-
thy)amino-2,4'-bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoropropa-
n-2-ol;
(2R)-3-(trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-
-4-yl)methyl)amino-2,4'-bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifl-
uoropropan-2-ol;
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(2-(trif-
luoromethoxy)ethylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((R)-2,2-dimethyltetrahydro-2-
H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((S)-2,2-dimethyltetrahydro-2-
H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-3,5,5'-trichloro-N6-((tetrahydro-2H-pyran-4-
-yl)methyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-((tetrahydro-2H-pyran-
-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((4-methyltetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-((4-methyltetrahydro--
2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((4-fluorotetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)cyc-
lohexanol;
5'-chloro-N2'-(trans-4-(dimethylamino)cyclohexyl)-N6-(3-fluorob-
enzyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-(2-methoxyethylamino)cyclohexy-
l)-2,4'-bipyridine-2',6-diamine;
2-(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)-
cyclohexylamino)ethanol;
5'-chloro-N6-(3,5-difluorobenzyl)-N2'-(trans-4-(2-methoxyethylamino)cyclo-
hexyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-((2-methoxyethyl)(methyl)amino-
)cyclohexyl)-2,4'-bipyridin-2,6-diamine;
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((2R,6S)-2,6-dimethyltetrahyd-
ro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
4-((5'-chloro-5-fluoro-2'-(trans-4-(2-methoxyethylamino)cyclohexylamino)--
2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile;
4-((2'-(trans-4-aminocyclohexylamino)-5'-chloro-5-fluoro-2,4'-bipyridin-6-
-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile;
4-((5'-chloro-5-fluoro-2'-(trans-4-(propylamino)cyclohexylamino)-2,4'-bip-
yridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile;
4-((5'-chloro-2'-(trans-4-(dipropylamino)cyclohexylamino)-5-fluoro-2,4'-b-
ipyridin-6-yl-amino)methyl)tetrahydro-2 H-pyran-4-carbonitrile;
4-((5'-chloro-5-fluoro-2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cycloh-
exylamino)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonit-
rile;
4-((5'-chloro-2'-(trans-4-((2-methyl-1,3-dioxolan-2-yl)methyl)aminoc-
yclohexylamino)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-car-
bonitrile;
(4-((5'-chloro-2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cycl-
ohexylamino)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-yl)met-
hanol; and
5'-chloro-5-fluoro-N6-((4-methyltetrahydro-2H-pyran-4-yl)methyl-
)-N2'-(trans-4-(1,1-dioxotetrahydrothiophen-3-yl-amino)cyclohexyl)-2,4'-bi-
pyridine-2',6-diamine.
9. The method of claim 1, wherein: R.sub.1 is X--R.sub.16; X is a
bond, or C.sub.1-2 alkyl; R.sub.16 is selected from the group
consisting of C.sub.1-2-alkyl, cyclopentyl, cyclohexyl, piperidine,
piperazine, morpholine, pyridine, pyrrolidine, cyclohexenyl, and
tetrahydro-2H-pyran; wherein R.sub.16 is substituted with one to
three groups selected from amino, hydroxyl, NHCH.sub.2-phenyl,
CH.sub.2-amino, COO-t-butyl, H, methoxy, NH--SO.sub.2-ethyl,
CH.sub.2--NHSO.sub.2-ethyl, SO.sub.2-ethyl, t-butyl, methyl,
CH.sub.2--COOH, CO--NHCH.sub.3, CON(CH.sub.3).sub.2,
NHC(CH.sub.3)--CH.sub.2--SO.sub.2--CH.sub.3,
NH--COO--CH.sub.2-phenyl, hydroxy-methyl, CH.sub.2--NH--CH.sub.3,
CH.sub.2--NH-ethyl, NH--CH.sub.2--CH.sub.2-methoxy,
CH.sub.2--NH--CO--CH.sub.3, NH--CH.sub.2--CH.sub.2OH,
NH--CO--CH.sub.2--N(CH.sub.3).sub.2, NH--CO-methylpyrrolidine,
NH--CH.sub.2--C(CH.sub.3)-dioxolane, NH--CO-pyridyl, NH-ethyl,
pyrrolidine, CH.sub.2--NH--CO-pyridyl, NH-tetrahydropyran,
COCH.sub.2--N(CH.sub.3).sub.2,
NH--CH.sub.2--C(CH.sub.3)-dimethyldioxolane, tetrahydropyran,
CO-methylpyrrolidine, CH.sub.2-methylpiperidine, NH--CO--CH.sub.3,
NH--SO.sub.2--CH.sub.3, NH--CH(CH.sub.2--OCH.sub.3).sub.2,
NH--CH.sub.2-tetrahydrofuran, NH--CH.sub.2-oxetane,
NH-tetrahydropyran, NH--CH.sub.2-dioxane,
N(CH.sub.3)--CH.sub.2CH.sub.2--OCH.sub.3, CH(OH)--CH.sub.2-amino,
NH--CH.sub.2CH.sub.2--OCF.sub.3, NHCH.sub.2--OCH.sub.3,
NH--CH.sub.2--CH(CF.sub.3)--OCH.sub.3,
NH--CH(CH.sub.3)--CH.sub.2--OH, F, NH-oxetane,
CH.sub.2--CH.sub.2--OCH.sub.3, CH.sub.2--OCH.sub.3,
CH.sub.2-tetrahydropyran, CH.sub.2-methylpiperizine,
NH.sub.2--CH.sub.2--CH(OH)--CF.sub.3, piperidine,
CH.sub.2-pyrrolidine, NH--CH(CH.sub.3)CH.sub.2OCH.sub.3,
NH-tetrahydrofuran, (CH.sub.2).sub.3--NH.sub.2, hydroxyethyl,
propyl, CH.sub.2-pyridyl, CH.sub.2-piperidine, morpholine,
NH-chloropyrimidine, NH--CH.sub.2CH.sub.2--SO.sub.2-methyl,
(CH.sub.3).sub.3--N(CH.sub.3).sub.2, piperizine, ##STR00511## and
CH.sub.2-morpholine; R.sub.3 is selected from H, methyl, cyano,
chloro, CONH.sub.2, amino, cyclopropyl, ethyl, and fluoro; R.sub.4
is selected from halogen, methyl, hydrogen, and halo-methyl;
R.sub.6 is H; R.sub.7 is selected from H, COOH, Cl, F, CONH.sub.2,
CN, and CF.sub.3; R.sub.8 is Cl; R.sub.17 and R.sub.18 are each,
independently, selected from the group consisting of hydrogen,
C.sub.1-3alkyl, C.sub.1-4haloalkyl, C.sub.3-6branched alkyl,
R.sub.22--OR.sub.12, R.sub.22--S(O).sub.2R.sub.12,
R.sub.22--NR.sub.15S(O).sub.2R.sub.12, heterocycloalkyl or
heteroaryl; alternatively, R.sub.17 and R.sub.18 along with the
nitrogen atom to which they are attached to can be taken together
to form a four to six membered heterocyclic ring wherein said ring
carbon atoms are optionally substituted with R.sub.20, and the ring
nitrogen atoms are optionally substituted with R.sub.21; R.sub.19
is selected from C.sub.1-3-alkyl, optionally substituted
heterocycloalkyl, optionally substituted aryl or optionally
substituted heteroaryl; R.sub.20 represents the group
C.sub.1-3alkyl; R.sub.22 is selection from the group consisting of
C.sub.1-4alkyl, and C.sub.3-6 branched alkyl; L is selected from
C.sub.1-4-alkyl, and cyclopropyl; R.sub.2 is selected from the
group consisting of C.sub.3-7 cycloalkyl, a five to seven membered
heterocycloalkyl, phenyl, and pyridyl, wherein each said R.sub.2
group is substituted with one, two, or three substituents
independently selected from hydrogen, cyano, CO--NH.sub.2, halogen,
methoxy, dihalo-methoxy, trihalo-methoxy, trihalo alkyl,
C.sub.1-3-alkyl, and hydroxy; and R.sub.9 represents methyl,
hydrogen, or ethyl.
10. The method of claim 1, wherein the compound is selected from:
3,5'-dichloro-N2'-(trans-4-(((R)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)am-
inocyclohexyl)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6--
diamine;
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cycloh-
exyl)-N6-(((S)-tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2',6-diami-
ne; 3,5'-di
chloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-(((R)--
tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2',6-diamine;
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-3-fluoro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexy-
l)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-5-fluoro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexy-
l)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-5-fluoro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-((1-
,1-dioxotetrahydro-2H-thiopyran-4-yl)methyl)aminocyclohexyl)-2,4'-bipyridi-
ne-2',6-diamine;
5'-chloro-5-fluoro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tet-
rahydro-2H-pyran-4;
3,5'-dichloro-N2'-(trans-A-(2-methoxyethylamino)cyclohexyl)-N6-((tetrahyd-
ro-2H-pyran-4-yl)methyl)-2,4-bipyridine-2',6-diamine;
2-(trans-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2,4'--
bipyridin-2'-yl-amino)cyclohexylamino)ethanol;
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((R)-te-
trahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine;
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((S)-te-
trahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine;
3,5'-dichloro-N2'-(trans-4-((2-methoxyethyl)(methyl)amino)cyclohexyl)-N6--
((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2,6-diamine;
5'-chloro-3-fluoro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tet-
rahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
3-bromo-5'-chloro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tetr-
ahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)am-
ino-2,4'-bipyridin-2'-yl-amino)cyclohexanol;
(2S)-3-(trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)m-
ethyl)amino-2,4'-bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoropro-
pan-2-ol;
(2R)-3-(trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyr-
an-4-yl)methyl)amino-2,4'-bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-tri-
fluoropropan-2-ol;
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(2-(trif-
luoromethoxy)ethylamino)cyclohexyl)-2,4'-bipyridin-2',6-diamine;
4-((5'-chloro-5-fluoro-2'-(trans-4-(2-methoxyethylamino)cyclohexylamino)--
2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile;
4-((2'-(trans-4-aminocyclohexylamino)-5'-chloro-5-fluoro-2,4'-bipyridin-6-
-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile;
4-((5'-chloro-5-fluoro-2'-(trans-4-(propylamino)cyclohexylamino)-2,4'-bip-
yridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile;
4-((5'-chloro-2'-(trans-4-(dipropylamino)cyclohexylamino)-5-fluoro-2,4'-b-
ipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile;
4-((5'-chloro-5-fluoro-2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cycloh-
exylamino)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonit-
rile; and
5'-chloro-5-fluoro-N6-((4-methyltetrahydro-2H-pyran-4-yl)methyl)-
-N2'-(trans-4-(1,1-dioxotetrahydrothiophen-3-yl-amino)cyclohexyl)-2,4'-bip-
yridine-2',6-diamine.
11. The method of claim 1, wherein the compound is selected from:
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)dideuteromethyl)-N2'-(trans-4-(((-
S)-tetrahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diami-
ne;
4-((5'-chloro-2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexylam-
ino)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile;
5'-chloro-N6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4--
((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-2,4-bipyridine-2',6-diamine;
5'-chloro-N6-(((S)-2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)-N2'-(tran-
s-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-2,4'-bipyridine-2',6-diami-
ne;
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((R)-tet-
rahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine;
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((S)-tetrah-
ydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine;
4-((5'-chloro-2'-(trans-4-((2-methyl-1,3-dioxolan-2-yl)methyl)aminocycloh-
exylamino)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonit-
rile; and
(4-((5'-chloro-2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclo-
hexylamino)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-yl)meth-
anol.
12. The method of claim 1, wherein the condition is selected from
cardiac hypertrophy, HIV and inflammatory diseases.
13. The method of claim 1, wherein the condition is a cancer is
selected from the group consisting of bladder, head and neck,
breast, stomach, ovary, colon, lung, brain, larynx, lymphatic
system, hematopoietic system, genitourinary tract,
gastrointestinal, ovarian, prostate, gastric, bone, small-cell
lung, glioma, colorectal, and pancreatic cancer.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of U.S. Ser.
No. 13/773,052, filed Feb. 21, 2013, which is a divisional
applicant of U.S. Ser. No. 12/843,494, filed Jul. 26, 2010, which
claims priority to U.S. provisional application Ser. No.
61/273,154, filed Jul. 30, 2009, and U.S. provisional application
Ser. No. 61/357,720, filed Jun. 23, 2010, the contents of which are
incorporated herein by reference in their entirety.
BACKGROUND
[0002] The search for new therapeutic agents has been greatly aided
in recent years by a better understanding of the structure of
enzymes and other biomolecules associated with diseases. One
important class of enzymes that has been the subject of extensive
study is protein kinases.
[0003] Protein kinases constitute a large family of structurally
related enzymes that are responsible for the control of a variety
of signal transduction processes within the cell. (Hardie, G. and
Hanks, S. The Protein Kinase Facts Book, I and II, Academic Press,
San Diego, Calif.: 1995). Protein kinases are thought to have
evolved from a common ancestral gene due to the conservation of
their structure and catalytic function. Almost all kinases contain
a similar 250-300 amino acid catalytic domain. The kinases may be
categorized into families by the substrates they phosphorylate
(e.g., protein-tyrosine, protein-serine/threonine, lipids, etc.).
Sequence motifs have been identified that generally correspond to
each of these kinase families (See, for example, Hanks, S. K.,
Hunter, T., FASEB J. 1995, 9, 576-596; Knighton et al., Science
1991, 253, 407-414; Hiles et al., Cell 1992, 70, 419-429; Kunz et
al., Cell 1993, 73, 585-596; Garcia-Bustos et al., EMBO J. 1994,
13, 2352-2361).
[0004] Many diseases are associated with abnormal cellular
responses triggered by the protein kinase-mediated events described
above. These diseases include, but are not limited to, autoimmune
diseases, inflammatory diseases, bone diseases, metabolic diseases,
neurological and neurodegenerative diseases, cancer, cardiovascular
diseases, allergies and asthma, Alzheimer's disease, viral
diseases, and hormone-related diseases. Accordingly, there has been
a substantial effort in medicinal chemistry to find protein kinase
inhibitors that are effective as therapeutic agents.
[0005] The cyclin-dependent kinase (CDK) complexes are a class of
kinases that are targets of interest. These complexes comprise at
least a catalytic (the CDK itself) and a regulatory (cyclin)
subunit. Some of the more important complexes for cell cycle
regulation include cyclin A (CDK1--also known as cdc2, and CDK2),
cyclin B1-B3 (CDK1) and cyclin DI-D3 (CDK2, CDK4, CDK5, CDK6),
cyclin E (CDK2). Each of these complexes is involved in a
particular phase of the cell cycle. Additionally, CDKs 7, 8, and 9
are implicated in the regulation of transcription.
[0006] The activity of CDKs is regulated post-translationally, by
transitory associations with other proteins, and by alterations of
their intracellular localization. Tumor development is closely
associated with genetic alteration and deregulation of CDKs and
their regulators, suggesting that inhibitors of CDKs may be useful
anti-cancer therapeutics. Indeed, early results suggest that
transformed and normal cells differ in their requirement for, e.g.,
cyclin A/CDK2 and that it may be possible to develop novel
antineoplastic agents devoid of the general host toxicity observed
with conventional cytotoxic and cytostatic drugs. While inhibition
of cell cycle-related CDKs is clearly relevant in, e.g., oncology
applications, inhibition of RNA polymerase-regulating CDKs may also
be highly relevant in cancer indications.
[0007] The CDKs have been shown to participate in cell cycle
progression and cellular transcription, and loss of growth control
is linked to abnormal cell proliferation in disease (see e.g.,
Malumbres and Barbacid, Nat. Rev. Cancer 2001, 1:222). Increased
activity or temporally abnormal activation of cyclin-dependent
kinases has been shown to result in the development of human tumors
(Sherr C. J., Science 1996, 274: 1672-1677). Indeed, human tumor
development is commonly associated with alterations in either the
CDK proteins themselves or their regulators (Cordon-Cardo C., Am.
J. Pat1/701. 1995; 147: 545-560; Karp J. E. and Broder S., Nat.
Med. 1995; 1: 309-320; Hall M. et al., Adv. Cancer Res. 1996; 68:
67-108).
[0008] Naturally occurring protein inhibitors of CDKs such as p16
and p27 cause growth inhibition in vitro in lung cancer cell lines
(Kamb A., Curr. Top. Microbiol. Immunol. 1998; 227: 139-148).
[0009] CDKs 7 and 9 seem to play key roles in transcription
initiation and elongation, respectively (see, e.g., Peterlin and
Price. Cell 23: 297-305, 2006, Shapiro. J. Clin. Oncol. 24:
1770-83, 2006). Inhibition of CDK9 has been linked to direct
induction of apoptosis in tumor cells of hematopoetic lineages
through down-regulation of transcription of antiapoptotic proteins
such as Mcl1 (Chao, S.- H. et al. J. Biol. Chem. 2000;
275:28345-28348; Chao, S.- H. et al. J. Biol. Chem. 2001;
276:31793-31799; Lam et. al. Genome Biology 2: 0041.1-11, 2001;
Chen et al. Blood 2005; 106:2513; MacCallum et al. Cancer Res.
2005; 65:5399; and Alvi et al. Blood 2005; 105:4484). In solid
tumor cells, transcriptional inhibition by downregulation of CDK9
activity synergizes with inhibition of cell cycle CDKs, for example
CDK1 and 2, to induce apoptosis (Cai, D.- P., Cancer Res 2006,
66:9270. Inhibition of transcription through CDK9 or CDK7 may have
selective non-proliferative effect on the tumor cell types that are
dependent on the transcription of mRNAs with short half lives, for
example Cyclin D1 in Mantle Cell Lymphoma. Some transcription
factors such as Myc and NF-kB selectively recruit CDK9 to their
promoters, and tumors dependent on activation of these signaling
pathways may be sensitive to CDK9 inhibition.
[0010] Small molecule CDK inhibitors may also be used in the
treatment of cardiovascular disorders such as restenosis and
atherosclerosis and other vascular disorders that are due to
aberrant cell proliferation. Vascular smooth muscle proliferation
and intimal hyperplasia following balloon angioplasty are inhibited
by over-expression of the cyclin-dependent kinase inhibitor
protein. Moreover, the purine CDK2 inhibitor CVT-313 (Ki=95 nM)
resulted in greater than 80% inhibition of neointima formation in
rats.
[0011] CDK inhibitors can be used to treat diseases caused by a
variety of infectious agents, including fungi, protozoan parasites
such as Plasmodium falciparum, and DNA and RNA viruses. For
example, cyclin-dependent kinases are required for viral
replication following infection by herpes simplex virus (HSV)
(Schang L. M. et al., J. Virol. 1998; 72: 5626) and CDK homologs
are known to play essential roles in yeast.
[0012] Inhibition of CDK9/cyclin T function was recently linked to
prevention of HIV replication and the discovery of new CDK biology
thus continues to open up new therapeutic indications for CDK
inhibitors (Sausville, E. A. Trends Molec. Med. 2002, 8,
S32-S37).
[0013] CDKs are important in neutrophil-mediated inflammation and
CDK inhibitors promote the resolution of inflammation in animal
models. (Rossi, A. G. et al, Nature Med. 2006, 12:1056). Thus CDK
inhibitors, including CDK9 inhibitors, may act as anti-inflammatory
agents.
[0014] Selective CDK inhibitors can be used to ameliorate the
effects of various autoimmune disorders. The chronic inflammatory
disease rheumatoid arthritis is characterized by synovial tissue
hyperplasia; inhibition of synovial tissue proliferation should
minimize inflammation and prevent joint destruction. In a rat model
of arthritis, joint swelling was substantially inhibited by
treatment with an adenovirus expressing a CDK inhibitor protein p
16. CDK inhibitors are effective against other disorders of cell
proliferation including psoriasis (characterized by keratinocyte
hyperproliferation), glomerulonephritis, chronic inflammation, and
lupus.
[0015] Certain CDK inhibitors are useful as chemoprotective agents
through their ability to inhibit cell cycle progression of normal
untransformed cells (Chen, et al. J. Natl. Cancer Institute, 2000;
92: 1999-2008). Pre-treatment of a cancer patient with a CDK
inhibitor prior to the use of cytotoxic agents can reduce the side
effects commonly associated with chemotherapy. Normal proliferating
tissues are protected from the cytotoxic effects by the action of
the selective CDK inhibitor.
[0016] Accordingly, there is a great need to develop inhibitors of
protein kinases, such as CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7,
CDK8 and CDK9, as well as combinations thereof.
SUMMARY OF THE INVENTION
[0017] There remains a need for new treatments and therapies for
protein kinase-associated disorders. There is also a need for
compounds useful in the treatment or prevention or amelioration of
one or more symptoms of cancer, inflammation, cardiac hypertrophy,
and HIV. Furthermore, there is a need for methods for modulating
the activity of protein kinases, such as CDK1, CDK2, CDK3, CDK4,
CDK5, CDK6, CDK7, CDK8 and CDK9, and combinations thereof, using
the compounds provided herein. In one aspect, the invention
provides a compound of Formula I:
##STR00002##
or a pharmaceutically acceptable salt thereof, wherein: A is N or
CR.sub.6;
A.sub.2 is N, N(O) or CR.sub.7;
A.sub.3 is N or CR.sub.8;
[0018] A.sub.4 is selected from a bond, SO.sub.2, NR.sub.9, or O; L
is selected from a bond, optionally substituted C.sub.1-4alkyl,
C.sub.3-6 cycloalkyl, C.sub.3-6 heterocycloalkyl, or C.sub.2-4
alkenyl;
R.sub.1 is X--R.sub.16;
[0019] X is a bond, or C.sub.1-4 alkyl and; R.sub.16 is selected
from the group consisting of C.sub.1-6 alkyl, C.sub.3-6 branched
alkyl, C.sub.3-6cycloalkyl, heterocycloalkyl, C.sub.3-8-partially
unsaturated cycloalkyl, aryl, and heteroaryl; wherein R.sub.16 is
substituted with one to three groups independently selected from
halogen, hydrogen, C.sub.1-6alkyl, C.sub.1-6haloalkyl,
C.sub.3-6branched alkyl, C.sub.3-6branched haloalkyl, OH,
C.sub.1-6alkoxy, R.sub.22--OR.sub.12, S(O).sub.0-2R.sub.12,
R.sub.22--S(O).sub.0-2R.sub.12, S(O).sub.2NR.sub.13R.sub.14,
R.sub.22--S(O).sub.2NR.sub.13R.sub.14, C(O)OR.sub.12,
R.sub.22--C(O)OR.sub.12, C(O)R.sub.19, R.sub.22--C(O)R.sub.19,
O--C.sub.1-3 alkyl, OC.sub.1-3 haloalkyl, OC(O)R.sub.19,
R.sub.22--OC(O)R.sub.19, C(O)NR.sub.13R.sub.14,
R.sub.22--C(O)NR.sub.13R.sub.14, NR.sub.15S(O).sub.2R.sub.12,
R.sub.22--NR.sub.15S(O).sub.2R.sub.12, NR.sub.17R.sub.18,
R.sub.22--NR.sub.17R.sub.18, NR.sub.15C(O)R.sub.19,
R.sub.22--NR.sub.15C(O)R.sub.19, NR.sub.15C(O)OCH.sub.2Ph,
R.sub.22--NR.sub.15C(O)OCH.sub.2Ph, NR.sub.15C(O)OR.sub.12,
R.sub.22--NR.sub.15C(O)OR.sub.12, NR.sub.15C(O)NR.sub.13R.sub.14,
and R.sub.22--NR.sub.15C(O)NR.sub.13R.sub.14; R.sub.17 and R.sub.18
are each, independently, selected from the group consisting of
hydrogen, hydroxyl, C.sub.1-6alkyl, C.sub.1-6haloalkyl,
C.sub.3-6branched alkyl, C.sub.3-6 cycloalkyl, R.sub.22--OR.sub.12,
R.sub.22--S(O).sub.0-2R.sub.12,
R.sub.22--S(O).sub.2NR.sub.13R.sub.14, R.sub.22--C(O)OR.sub.12,
R.sub.22--C(O)R.sub.19, R.sub.22--OC(O)R.sub.19,
R.sub.22--C(O)NR.sub.13R.sub.14,
R.sub.22--NR.sub.15S(O).sub.2R.sub.12, R.sub.22--NR.sub.23R.sub.24,
R.sub.22--NR.sub.15C(O)R.sub.19,
R.sub.22--NR.sub.15C(O)OCH.sub.2Ph,
R.sub.22--NR.sub.15C(O)OR.sub.12,
R.sub.22--NR.sub.15C(O)NR.sub.13R.sub.14, cycloalkyl,
heterocycloalkyl and heteroaryl; alternatively, R.sub.17 and
R.sub.18 along with the nitrogen atom to which they are attached to
can be taken together to form a four to six membered heterocyclic
ring wherein the carbon atoms of said ring are optionally
substituted with R.sub.20, and the nitrogen atoms of said ring are
optionally substituted with R.sub.21; R.sub.19 is selected from
optionally substituted alkyl, optionally substituted cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, and optionally substituted heteroaryl; R.sub.20 is selected
from the group consisting of C.sub.1-6 alkyl or C.sub.1-6
haloalkyl; R.sub.21 is selected from the group consisting of
C.sub.1-6alkyl, C.sub.1-6haloalkyl, C(O)R.sub.12, C(O)OR.sub.12,
S(O).sub.2R.sub.12; R.sub.22 is selected from the group consisting
of C.sub.1-6 alkyl, C.sub.1-6haloalkyl, C.sub.3-6 branched alkyl,
C.sub.3-6branched haloalkyl;
[0020] R.sub.23 and R.sub.24 are each, independently, selected from
the group consisting of hydrogen, C.sub.1-6 alkyl,
C.sub.1-6haloalkyl, C.sub.3-6 branched alkyl, C.sub.3-6 branched
haloalkyl;
R.sub.2 is selected from the group consisting of optionally
substituted C.sub.1-6 alkyl, optionally substituted C.sub.3-6
branched alkyl, optionally substituted C.sub.3-8 cycloalkyl,
optionally substituted heterocycloalkyl, optionally substituted
aryl, and optionally substituted heteroaryl; R.sub.4, R.sub.5, and
R.sub.6 are each, independently, selected from the group consisting
of hydrogen, hydroxyl, cyano, halogen, C.sub.1-4 alkyl,
C.sub.1-4haloalkyl, C.sub.2-4 alkenyl, C.sub.2-4 alkynyl, amino,
NR.sub.10R.sub.11, and alkoxy; R.sub.3, R.sub.7 and R.sub.8 are
each, independently, selected from the group consisting of
hydrogen, hydroxyl, cyano, halogen, alkyl, haloalkyl, alkenyl,
alkynyl, alkoxy, NR.sub.10R.sub.11, C(O)R.sub.12, C(O)OR.sub.12,
C(O)NR.sub.13R.sub.14, S(O).sub.0-2R.sub.12,
S(O).sub.0-2NR.sub.13R.sub.14, and optionally substituted C.sub.3-4
cycloalkyl; R.sub.9 is selected from the group consisting of
hydrogen, C.sub.1-4 alkyl, alkoxy, C(O)R.sub.12, C(O)OR.sub.15,
C(O)NR.sub.13R.sub.14, S(O).sub.0-2R.sub.12, S(O).sub.0-2
NR.sub.13R.sub.14, optionally substituted C.sub.3-4 cycloalkyl, and
optionally substituted heterocycloalkyl; R.sub.10 and R.sub.11 are
each, independently, selected from the group consisting of
hydrogen, hydroxyl, alkyl, alkoxy, C(O)R.sub.12, C(O)OR.sub.12,
C(O)NR.sub.13R.sub.14, S(O).sub.0-2R.sub.12, and
S(O).sub.0-2NR.sub.13R.sub.14; alternatively, R.sub.11 and R.sub.12
along with the nitrogen atom to which they are attached to can be
taken together to form an optionally substituted four to six
membered heteroaromatic, or a non-aromatic heterocyclic ring;
[0021] R.sub.12 and R.sub.15 are each, individually, selected from
the group consisting of hydrogen, alkyl, branched alkyl, haloalkyl,
branched haloalkyl, (CH.sub.2).sub.0-3-cycloalkyl,
(CH.sub.2).sub.0-3-heterocycloalkyl, (CH.sub.2).sub.0-3-aryl, and
heteroaryl;
R.sub.13 and R.sub.14 are each, independently, selected from the
group consisting of hydrogen, hydroxyl, alkyl, branched alkyl,
haloalkyl, branched haloalkyl, alkoxy, cycloalkyl or
heterocycloalkyl; and alternatively, R.sub.13 and R.sub.14 along
with the nitrogen atom to which they are attached to can be taken
together to form an optionally substituted four to six membered
heteroaromatic, or non-aromatic heterocyclic ring.
[0022] One preferred embodiment of the present invention provides a
compound of Formula I wherein, A.sub.1 is N; A.sub.2 is N; and
A.sub.3 is CR.sub.8. Another aspect of the present invention
provides a compound of Formula I wherein A.sub.1 is CR.sub.6,
A.sub.2 is CR.sub.7, and A.sub.3 is CR.sub.8. Yet another preferred
embodiment provides a compound of Formula I wherein, A.sub.1 is N;
A.sub.2 is CR.sub.7; and A.sub.3 is CR.sub.8. A further preferred
embodiment of the preceding aspects of the present invention
provides a compound of Formula I wherein, R.sub.8 is selected from
halogen, hydrogen, CN, CF.sub.3, O--C.sub.1-3-alkyl, and
C.sub.1-3-alkyl, with Cl, F, and methyl being the preferred R.sub.8
substituents, and Cl being the particularly preferred R.sub.8
substituent. Other preferred embodiment of the present invention
provides a compound of Formula I wherein, R.sub.1 is X--R.sub.16; X
is a bond, or C.sub.1-2 alkyl; R.sub.16 is selected from the group
consisting of C.sub.1-2-alkyl, C.sub.4-6cycloalkyl,
heterocycloalkyl, phenyl, and heteroaryl; wherein R.sub.16 is
substituted with one to three groups independently selected from
halogen, hydrogen, C.sub.1-3alkyl, C.sub.3-6branched alkyl, OH,
C.sub.1-2alkoxy, R.sub.22--OR.sub.12, S(O).sub.1-2R.sub.12,
C(O)OR.sub.12, R.sub.22--C(O)OR.sub.12, C(O)R.sub.19,
R.sub.22--OC(O)R.sub.19, C(O)NR.sub.13R.sub.14,
NR.sub.15S(O).sub.2R.sub.12, NR.sub.17R.sub.18,
R.sub.22--NR.sub.17R.sub.18, NR.sub.15C(O)R.sub.19,
R.sub.22--NR.sub.15C(O)R.sub.19, and NR.sub.15C(O)OCH.sub.2Ph.
Particularly preferred R.sub.16 substituents are selected from the
group consisting of C.sub.1-2-alkyl, cyclopentyl, cyclohexyl,
piperidine, piperazine, morpholine, pyridine, pyrrolidine,
cyclohexenyl, and tetrahydro-2H-pyran; wherein R.sub.16 is
substituted with one to three groups selected from amino, hydroxyl,
NHCH.sub.2-phenyl, CH.sub.2-amino, COO-t-butyl, H, methoxy,
NH--SO.sub.2-ethyl, CH.sub.2--NHSO.sub.2-ethyl, SO.sub.2-ethyl,
t-butyl, methyl, CH.sub.2--COOH, CO--NHCH.sub.3,
CON(CH.sub.3).sub.2, NHC(CH.sub.3)--CH.sub.2--SO.sub.2--CH.sub.3,
NH--COO--CH.sub.2-phenyl, hydroxy-methyl, CH.sub.2--NH--CH.sub.3,
CH.sub.2--NH-ethyl, NH--CH.sub.2--CH.sub.2-methoxy,
CH.sub.2--NH--CO--CH.sub.3, NH--CH.sub.2--CH.sub.2OH,
NH--CO--CH.sub.2--N(CH.sub.3).sub.2, NH--CO-methylpyrrolidine,
NH--CH.sub.2--C(CH.sub.3)-dioxolane, NH--CO-pyridyl, NH-ethyl,
pyrrolidine, CH.sub.2--NH--CO-pyridyl, NH-tetrahydropyran,
COCH.sub.2--N(CH.sub.3).sub.2,
NH--CH.sub.2--C(CH.sub.3)-dimethyldioxolane, tetrahydropyran,
CO-methylpyrrolidine, CH.sub.2-methylpiperidine, NH--CO--CH.sub.3,
NH--SO.sub.2--CH, NH--CH(CH.sub.2--OCH.sub.3).sub.2,
NH--CH.sub.2-tetrahydrofuran, NH--CH.sub.2-oxetane,
NH-tetrahydropyran, NH--CH.sub.2-dioxane,
N(CH.sub.3)--CH.sub.2CH.sub.2--OCH.sub.3, CH(OH)--CH.sub.2-amino,
NH--CH.sub.2CH.sub.2--OCF.sub.3, NHCH.sub.2--OCH.sub.3,
NH--CH.sub.2--CH(CF.sub.3)--OCH.sub.3,
NH--CH(CH.sub.3)--CH.sub.2--OH, F, NH-oxetane,
CH.sub.2--CH.sub.2--OCH.sub.3, CH.sub.2--OCH.sub.3,
CH.sub.2-tetrahydropyran, CH.sub.2-methylpiperizine,
NH.sub.2--CH.sub.2--CH(OH)--CF.sub.3, piperidine,
CH.sub.2-pyrrolidine, NH--CH(CH.sub.3)CH.sub.2OCH.sub.3,
NH-tetrahydrofuran, (CH.sub.2).sub.3--NH.sub.2, hydroxyethyl,
propyl, CH.sub.2-pyridyl, CH.sub.2-piperidine, morpholine,
NH-chloropyrimidine, NH--CH.sub.2CH.sub.2--SO.sub.2-methyl,
(CH.sub.3).sub.3--N(CH.sub.3).sub.2, piperizine,
##STR00003##
and CH.sub.2-morpholine.
[0023] Another preferred embodiment of the present invention
provides a compound of Formula I wherein, R.sub.3 is selected from
H, methyl, cyano, chloro, CONH.sub.2, amino, cyclopropyl, ethyl,
and fluoro; R.sub.4 is selected from halogen, methyl, hydrogen, and
halo-methyl; R.sub.6 is H; R.sub.7 is selected from H, COOH, Cl, F,
CONH.sub.2, CN, and CF.sub.3; R.sub.8 is Cl; R.sub.17 and R.sub.18
are each, independently, selected from the group consisting of
hydrogen, C.sub.1-3alkyl, C.sub.1-4haloalkyl, C.sub.3-6branched
alkyl, R.sub.22--OR.sub.12, R.sub.22--S(O).sub.2R.sub.12,
R.sub.22--NR.sub.15S(O).sub.2R.sub.12, heterocycloalkyl or
heteroaryl; alternatively, R.sub.17 and R.sub.18 along with the
nitrogen atom to which they are attached to can be taken together
to form a four to six membered heterocyclic ring wherein said ring
carbon atoms are optionally substituted with R.sub.20, and the ring
nitrogen atoms are optionally substituted with R.sub.21;
[0024] R.sub.19 is selected from C.sub.1-3-alkyl, optionally
substituted heterocycloalkyl, optionally substituted aryl or
optionally substituted heteroaryl; R.sub.20 represents the group
C.sub.1-3alkyl; and R.sub.22 is selection from the group consisting
of C.sub.1-4alkyl, and C.sub.3-6 branched alkyl. Further preferred
are compounds of Formula I wherein, A.sub.4 is selected from
NR.sub.9, O, and a bond; L is selected from a bond,
C.sub.1-4-alkyl, and cyclopropyl; R.sub.2 is selected from the
group consisting of C.sub.3-7 cycloalkyl, a five to seven membered
heterocycloalkyl, phenyl, and pyridyl, wherein each said R.sub.2
group is substituted with one, two, or three substituents
independently selected from hydrogen, cyano, CO--NH.sub.2, halogen,
methoxy, dihalo-methoxy, trihalo-methoxy, trihalo alkyl,
C.sub.1-3-alkyl, and hydroxy; and R.sub.9 represents methyl,
hydrogen, or ethyl.
[0025] Provided in a particularly preferred embodiment are
compounds of Formula I, wherein, A.sub.1 is CR.sub.6; A.sub.2 is
CR.sub.7; A.sub.3 is CR.sub.8; A.sub.4 is selected from NR.sub.9,
O, and a bond; L is a bond, C.sub.1-2 alkyl, or
C.sub.3-4-cycloalkyl; R.sub.1 is X--R.sub.16; X is a bond, or
C.sub.1-2 alkyl; R.sub.16 is selected from the group consisting of
C.sub.1-2-alkyl, cyclopentyl, cyclohexyl, piperidine, piperazine,
morpholine, pyridine, pyrrolidine, cyclohexenyl, and
tetrahydro-2H-pyran; wherein R.sub.16 is substituted with one to
three groups independently selected from amino, hydroxyl,
NHCH.sub.2-phenyl, CH.sub.2-amino, COO-t-butyl, H, methoxy,
NH--SO.sub.2-ethyl, CH.sub.2--NHSO.sub.2-ethyl, SO.sub.2-ethyl,
t-butyl, methyl, CH.sub.2--COOH, CO--NHCH.sub.3,
CON(CH.sub.3).sub.2, NHCH.sub.2--CH.sub.2--SO.sub.2--CH.sub.3,
NH--COO--CH.sub.2-phenyl, hydroxy-methyl, CH.sub.2--NH--CH.sub.3,
CH.sub.2--NH-ethyl, NH--CH.sub.2--CH.sub.2-methoxy,
CH.sub.2--NH--CO--CH.sub.3, NH--CH.sub.2--CH.sub.2OH,
NH--CO--CH.sub.2--N(CH.sub.3).sub.2, NH--CO-methylpyrrolidine,
NH--CO-pyridyl, NH-ethyl, pyrrolidine, CH.sub.2--NH--CO-pyridyl,
COCH.sub.2--N(CH.sub.3).sub.2, tetrahydropyran,
CO-methylpyrrolidine, CH.sub.2-methylpiperidine, NH--CO--CH.sub.3,
NH--SO.sub.2--CH.sub.3, NH--CH.sub.2-tetrahydrofuran,
NH--CH.sub.2-dioxane, N(CH.sub.3)--CH.sub.2CH.sub.2--OCH.sub.3,
CH(OH)--CH.sub.2-amino, NH--CH.sub.2CH.sub.2--OCF.sub.3,
NH(CH.sub.3)--CH.sub.2--OCH.sub.3,
NH--CH.sub.2--CH(CF.sub.3)--OCH.sub.3, F, NH-oxetane,
CH.sub.2--CH.sub.2--OCH.sub.3, CH.sub.2--OCH.sub.3,
CH.sub.2-tetrahydropyran, CH.sub.2-methylpiperizine,
NH.sub.2--CH.sub.2--CH(OH)--CF.sub.3, piperidine,
CH.sub.2-pyrrolidine, NH--CH(CH.sub.3)CH.sub.2OCH.sub.3,
NH-tetrahydrofuran, (CH.sub.2).sub.3--NH.sub.2, hydroxyethyl,
propyl, CH.sub.2-pyridyl, CH.sub.2-piperidine, morpholine,
NH-chloropyrimidine, NH--CH.sub.2CH.sub.2--SO.sub.2-methyl,
(CH.sub.3).sub.3--N(CH.sub.3).sub.2, piperizine,
CH.sub.2-morpholine, NH--CH.sub.2--C(CH.sub.3)-dioxolane,
NH-tetrahydropyran, NH--CH.sub.2--C(CH.sub.3)-dimethyldioxolane,
NH--CH(CH.sub.2--OCH.sub.3).sub.2, NH--CH.sub.2-oxetane,
NH-tetrahydropyran, N(CH.sub.3)--CH.sub.2CH.sub.2--OCH.sub.3,
NH--CH(CH.sub.3)--CH.sub.2--OH,
##STR00004##
[0026] and NH--CH(CH.sub.3)--CH.sub.2--OH;
[0027] R.sub.2 is selected from the group consisting of cyclohexyl,
1,3-dioxane, pyridinyl, phenyl, tetrahydropyranyl, cycloheptyl,
1,4-dioxane, morpholinyl, alkyl substituted dioxane,
tetrahydrofuranyl, dioxepane, piperidinyl, and
##STR00005##
[0028] wherein each said R.sub.2 group is substituted with one,
two, or three substituents independently selected from Cl, Br, F,
methoxy, hydroxy-methyl, hydrogen, carboxamide, cyano,
dihalo-methoxy, trihalo-methoxy, trifluoro-methyl, hydroxyl, and
methyl; and
[0029] R.sub.4, is chloro, hydrogen, trifluoro-methyl, fluoro, or
bromo;
[0030] R.sub.5, and R.sub.6 are each independently hydrogen;
[0031] R.sub.3 is selected from hydrogen, fluoro, cyano,
CO--NH.sub.2, chloro, amino, methyl, and cyclopropyl;
[0032] R.sub.7 is selected from H, trifluoro-methyl, COOH,
CO--NH.sub.2, and cyano;
[0033] R.sub.8 represents Cl; and
[0034] R.sub.9 is selected from the group consisting of H, ethyl,
and methyl.
[0035] Provided in a specifically preferred embodiment of the
present invention is a compound of Formula I selected from: [0036]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-3,5-difluoro-N6-((tetrahydro-2H-p-
yran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0037]
N2'-(trans-4-aminocyclohexyl)-5'-fluoro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine; [0038]
3,5'-dichloro-N2'-(trans-4-(((R)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)am-
inocyclohexyl)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6--
diamine; [0039]
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)dideuteromethyl)-N2'-(trans-4-(((-
S)-tetrahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diami-
ne; [0040]
5'-chloro-5-fluoro-N2'-(trans-4-(2-(methylsulfonyl)ethylamino)c-
yclohexyl)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diam-
ine; [0041]
5'-chloro-5-fluoro-N2'-(trans-4-(oxetan-2-yl-methylamino)cyclohexyl)-N6-(-
(tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
[0042]
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-(((S)-tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2',6-diamine;
[0043]
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohe-
xyl)-N6-(((R)-tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2',6-diamin-
e; [0044]
4-((5'-chloro-2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cycloh-
exylamino)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonit-
rile; [0045]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-(3-fluorobenzyl)-2,4'-
-bipyridine-2',6-diamine; [0046]
2'-(trans-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'--
bipyridine-5-carbonitrile; [0047]
N2'-(trans-4-aminocyclohexyl)-3-chloro-5'-fluoro-N6-((tetrahydro-2H-pyran-
-4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0048]
5'-chloro-N6-(3-fluorobenzyl)-N2'-((1R,3S)-3-((methylamino)methyl)cyclope-
ntyl)-2,4'-bipyridine-2',6-diamine; [0049]
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
[0050]
5'-chloro-3-fluoro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexy-
l)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
[0051]
5'-chloro-5-fluoro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cy-
clohexyl)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diami-
ne; [0052]
5'-chloro-N6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)-N2'-
-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-2,4'-bipyridine-2',6-
-diamine; [0053]
5'-chloro-N6-(((S)-2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)-N2'-(tran-
s-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-2,4'-bipyridine-2',6-diami-
ne; [0054]
5'-chloro-5-fluoro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(t-
rans-4-((1,1-dioxotetrahydro-2H-thiopyran-4-yl)methyl)aminocyclohexyl)-2,4-
'-bipyridine-2',6-diamine; [0055]
5'-chloro-5-fluoro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tet-
rahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0056]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine; [0057]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3,5-difluorobenzyl)-2,4'-bipy-
ridine-2',6-diamine; [0058]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((5-fluoropyridin-3-yl)methyl)-
-2,4'-bipyridine-2',6-diamine; [0059]
trans-4-(5'-chloro-6-(3,5-difluorobenzylamino)-2,4'-bipyridin-2'-yl-amino-
)cyclohexanol; [0060]
(R)-5'-chloro-N6-(3-fluorobenzyl)-N2'-(2-(piperidin-3-yl)ethyl)-2,4'-bipy-
ridine-2',6-diamine; [0061]
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine; [0062]
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine; [0063]
3,5'-dichloro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tetrahyd-
ro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0064]
2-(trans-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2,4'--
bipyridin-2'-yl-amino)cyclohexylamino)ethanol; [0065]
trans-N1-(5-chloro-4-(6-(((R)-tetrahydro-2H-pyran-3-yl)methyl)aminopyrazi-
n-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine; [0066]
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((R)-te-
trahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine;
[0067]
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(-
((S)-tetrahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-dia-
mine; [0068]
3,5'-dichloro-N2'-(trans-4-((2-methoxyethyl)(methyl)amino)cyclohexyl)-N6--
((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine;
[0069]
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((R)-tetrah-
ydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine;
[0070]
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((S)-
-tetrahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine-
; [0071]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-3-fluoro-N6-((tetrahydro--
2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0072]
5'-chloro-3-fluoro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tet-
rahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0073]
N2'-(trans-4-aminocyclohexyl)-3-bromo-5'-chloro-N6-((tetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0074]
3-bromo-5'-chloro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tetr-
ahydro-2H-pyran-4-yl)methyl)-2,4-bipyridine-2',6-diamine; [0075]
trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)am-
ino-2,4'-bipyridin-2'-yl-amino)cyclohexanol; [0076]
(2S)-3-(trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)m-
ethyl)amino-2,4'-bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoropro-
pan-2-ol; [0077]
(2R)-3-(trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)m-
ethyl)amino-2,4'-bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoropro-
pan-2-ol; [0078]
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(2-(trif-
luoromethoxy)ethylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine;
[0079]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((R)-2,2-dimethyltetrahydro-2-
H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0080]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((S)-2,2-dimethyltetrahydro-2-
H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0081]
N2'-(trans-4-aminocyclohexyl)-3,5,5'-trichloro-N6-((tetrahydro-2H-pyran-4-
-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0082]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-((tetrahydro-2H-pyran-
-4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0083]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((4-methyltetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0084]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-((4-methyltetrahydro--
2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0085]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((4-fluorotetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0086]
trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)cyc-
lohexanol; [0087]
5'-chloro-N2'-(trans-4-(dimethylamino)cyclohexyl)-N6-(3-fluorobenzyl)-2,4-
'-bipyridine-2',6-diamine; [0088]
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-(2-methoxyethylamino)cyclohexy-
l)-2,4'-bipyridine-2',6-diamine; [0089]
2-(trans-4-(5'-chloro-6-(3-fluorobenzyl
amino)-2,4'-bipyridin-2'-yl-amino)cyclohexylamino)ethanol; [0090]
5'-chloro-N6-(3,5-difluorobenzyl)-N2'-(trans-4-(2-methoxyethylamino)cyclo-
hexyl)-2,4'-bipyridine-2',6-diamine; [0091]
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-((2-methoxyethyl)(methyl)amino-
)cyclohexyl)-2,4'-bipyridine-2',6-diamine; [0092]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((2R,6S)-2,6-dimethyltetrahyd-
ro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine; [0093]
((5'-chloro-5-fluoro-2'-(trans-4-(2-methoxyethylamino)cyclohexylamino)-2,-
4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile;
[0094]
((2'-(trans-4-aminocyclohexylamino)-5'-chloro-5-fluoro-2,4'-bipyridin-6-y-
l-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile; [0095]
((5'-chloro-5-fluoro-2'-(trans-4-(propylamino)cyclohexylamino)-2,4'-bipyr-
idin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile; [0096]
((5'-chloro-2'-(trans-4-(dipropylamino)cyclohexylamino)-5-fluoro-2,4'-bip-
yridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile; [0097]
((5'-chloro-5-fluoro-2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohex-
ylamino)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitri-
le; [0098]
((5'-chloro-2'-(trans-4-((2-methyl-1,3-dioxolan-2-yl)methyl)ami-
nocyclohexylamino)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-ca-
rbonitrile; [0099]
(4-((5'-chloro-2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexylamin-
o)-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-pyran-4-yl)methanol;
and [0100]
5'-chloro-5-fluoro-N6-((4-methyltetrahydro-2H-pyran-4-yl)methyl)-N-
2'-(trans-4-(1,1-dioxotetrahydrothiophen-3-yl-amino)cyclohexyl)-2,4'-bipyr-
idine-2',6-diamine. [0101] Yet another preferred embodiment of the
present invention provides a compound of Formula I selected
from:
[0102]
trans-N1-(4-(3-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyra-
zin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine; [0103]
trans-N1-(5-chloro-4-(3-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminop-
yrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine; [0104]
trans-4-(5-chloro-4-(5-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopy-
razin-2-yl)pyridin-2-yl-amino)cyclohexanol; [0105]
trans-N1-(5-chloro-4-(5-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminop-
yrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine; [0106]
trans-4-(5-chloro-4-(6-(((S)-tetrahydro-2H-pyran-3-yl)methyl)aminopyrazin-
-2-yl)pyridin-2-yl-amino)cyclohexanol; [0107]
trans-4-(5-chloro-4-(6-(((R)-tetrahydro-2H-pyran-3-yl)methyl)aminopyrazin-
-2-yl)pyridin-2-yl-amino)cyclohexanol; [0108]
trans-N1-(5-chloro-4-(6-(((S)-tetrahydro-2H-pyran-3-yl)methyl)aminopyrazi-
n-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine; [0109]
trans-N1-(5-chloro-4-(6-(((R)-tetrahydro-2H-pyran-3-yl)methyl)aminopyrazi-
n-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine; [0110]
trans-N1-(5-chloro-4-(6-(methyl((tetrahydro-2H-pyran-4-yl)methyl)amino)py-
razin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine; [0111]
trans-N1-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyrazin-2--
yl)pyridin-2-yl)-N4-(2-methoxyethyl)cyclohexane-1,4-diamine; [0112]
trans-4-(5-chloro-4-(6-((tetrahydro-2H-pyran-3-yl)methyl)aminopyrazin-2-y-
l)pyridin-2-yl-amino)cyclohexanol; [0113]
trans-4-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyrazin-2-y-
l)pyridin-2-yl-amino)cyclohexanol; and [0114]
trans-N1-(5-chloro-4-(6-(3-fluorobenzylamino)pyrazin-2-yl)pyridin-2-yl)cy-
clohexane-1,4-diamine.
[0115] Another aspect of the present invention provides a compound
of Formula I, or pharmaceutically acceptable salt or solvate
thereof, for use in therapy. Yet another aspect of the present
invention provides a compound of Formula I or a pharmaceutically
acceptable salt or solvate thereof for use in a method of treating
a disease or condition mediated by CDK9.
[0116] Yet another aspect of the present invention provides a
method of treating a disease or condition mediated by CDK9
comprising administration to a subject in need thereof a
therapeutically effective amount of a compound of Formula I, or a
pharmaceutically acceptable salt thereof. Provided in yet another
aspect of the present invention is a compound of Formula I for use
in a method of treating a disease or condition mediated by CDK9 is
selected from cancer, cardiac hypotrophy, HIV and inflammatory
diseases.
[0117] Another aspect of the present invention provides a method of
treating a cancer selected from the group consisting of bladder,
head and neck, breast, stomach, ovary, colon, lung, brain, larynx,
lymphatic system, hematopoetic system, genitourinary tract,
gastrointestinal, ovarian, prostate, gastric, bone, small-cell
lung, glioma, colorectal. and pancreatic cancer.
[0118] Yet another aspect of the present invention provides a
pharmaceutical composition comprising a compound of Formula I, r a
pharmaceutically acceptable salt thereof, and a pharmaceutically
acceptable carrier, diluent or excipient.
[0119] In another aspect, the invention provides a method of
regulating, modulating, or inhibiting protein kinase activity which
comprises contacting a protein kinase with a compound of the
invention. In one embodiment, the protein kinase is selected from
the group consisting of CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7,
CDK8 and CDK9, or any combination thereof. In another embodiment,
the protein kinase is selected from the group consisting of CDK1,
CDK2 and CDK9, or any combination thereof. In still another
embodiment, the protein kinase is in a cell culture. In yet another
embodiment, the protein kinase is in a mammal.
[0120] In another aspect, the invention provides a method of
treating a protein kinase-associated disorder comprising
administering to a subject in need thereof a pharmaceutically
acceptable amount of a compound of the invention such that the
protein kinase-associated disorder is treated. In one embodiment,
the protein kinase is selected from the group consisting of CDK1,
CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8 and CDK9.
[0121] In one embodiment, the protein kinase-associated disorder is
cancer. In still another embodiment, the cancer is selected from
the group consisting of bladder, head and neck, breast, stomach,
ovary, colon, lung, brain, larynx, lymphatic system, hematopoetic
system, genitourinary tract, gastrointestinal, ovarian, prostate,
gastric, bone, small-cell lung, glioma, colorectal and pancreatic
cancer.
[0122] In one embodiment, the protein kinase-associated disorder is
inflammation. In another embodiment, the inflammation is related to
rheumatoid arthritis, lupus, type 1 diabetes, diabetic nephropathy,
multiple sclerosis, glomerulonephritis, chronic inflammation, and
organ transplant rejections.
[0123] In another embodiment, the protein kinase-associated
disorder is a viral infection. In one embodiment, the viral
infection is associated with the HIV virus, human papilloma virus,
herpes virus, poxyirus virus, Epstein-Barr virus, Sindbis virus, or
adenovirus.
[0124] In still another embodiment, the protein kinase-associated
disorder is cardiac hypertrophy.
[0125] In another aspect, the invention provides a method of
treating cancer comprising administering to a subject in need
thereof a pharmaceutically acceptable amount of a compound of the
invention such that the cancer is treated. In one embodiment, the
cancer is selected from the group consisting of bladder, head and
neck, breast, stomach, ovary, colon, lung, brain, larynx, lymphatic
system, hematopoetic system, genitourinary tract, gastrointestinal,
ovarian, prostate, gastric, bone, small-cell lung, glioma,
colorectal and pancreatic cancer.
[0126] In another aspect, the invention provides a method of
treating inflammation comprising administering to a subject in need
thereof a pharmaceutically acceptable amount of a compound such
that the inflammation is treated, wherein the compound is a
compound of the invention. In one embodiment, the inflammation is
related to rheumatoid arthritis, lupus, type 1 diabetes, diabetic
nephropathy, multiple sclerosis, glomerulonephritis, chronic
inflammation, and organ transplant rejections.
[0127] In another aspect, the invention provides a method of
treating cardiac hypertrophy comprising administering to a subject
in need thereof a pharmaceutically acceptable amount of a compound
such that the cardiac hypertrophy is treated, wherein the compound
is a compound of the invention.
[0128] In another aspect, the invention provides a method of
treating a viral infection comprising administering to a subject in
need thereof a pharmaceutically acceptable amount of a compound
such that the viral infection is treated, wherein the compound is a
compound of the invention. In one embodiment, the viral infection
is associated with the HIV virus, human papilloma virus, herpes
virus, poxyirus virus, Epstein-Barr virus, Sindbis virus, or
adenovirus.
[0129] In one embodiment, the subject to be treated by the
compounds of the invention is a mammal. In another embodiment, the
mammal is a human.
[0130] In another aspect, the compounds of the invention is
administered, simultaneously or sequentially, with an
antiinflammatory, antiproliferative, chemotherapeutic agent,
immunosuppressant, anti-cancer, cytotoxic agent or kinase inhibitor
or salt thereof. In one embodiment, the compound, or salt thereof,
is administered, simultaneously or sequentially, with one or more
of a PTK inhibitor, cyclosporin A, CTLA4-Ig, antibodies selected
from anti-ICAM-3, anti-IL-2 receptor, anti-CD45RB, anti-CD2,
anti-CD3, anti-CD4, anti-CD80, anti-CD86, and monoclonal antibody
OKT3, CVT-313, agents blocking the interaction between CD40 and
gp39, fusion proteins constructed from CD40 and gp39, inhibitors of
NF-kappa B function, non-steroidal antiinflammatory drugs,
steroids, gold compounds, FK506, mycophenolate mofetil, cytotoxic
drugs, TNF-.alpha. inhibitors, anti-TNF antibodies or soluble TNF
receptor, rapamycin, leflunimide, cyclooxygenase-2 inhibitors,
paclitaxel, cisplatin, carboplatin, doxorubicin, carminomycin,
daunorubicin, aminopterin, methotrexate, methopterin, mitomycin C,
ecteinascidin 743, porfiromycin, 5-fluorouracil, 6-mercaptopurine,
gemcitabine, cytosine arabinoside, podophyllotoxin, etoposide,
etoposide phosphate, teniposide, melphalan, vinblastine,
vincristine, leurosidine, epothilone, vindesine, leurosine, or
derivatives thereof.
[0131] In another aspect, the invention provides a packaged protein
kinase-associated disorder treatment, comprising a protein
kinase-modulating compound of the Formula I or Formula II, packaged
with instructions for using an effective amount of the protein
kinase-modulating compound to treat a protein kinase-associated
disorder.
[0132] In certain embodiments, the compound of the present
invention is further characterized as a modulator of a protein
kinase, including, but not limited to, protein kinases selected
from the group consisting of abl, ATK, Bcr-abl, Blk, Brk, Btk,
c-fms, c-kit, c-met, c-src, CDK, cRaf1, CSFIR, CSK, EGFR, ErbB2,
ErbB3, ErbB4, ERK, Fak, fes, FGFRI, FGFR2, FGFR3, FGFR4, FGFR5,
Fgr, FLK-4, flt-1, Fps, Frk, Fyn, GSK, Gst-Flk1, Hck, Her-2, Her-4,
IGF-1R, INS-R, Jak, JNK, KDR, Lck, Lyn, MEK, p38, panHER, PDGFR,
PLK, PKC, PYK2, Raf, Rho, ros, SRC, TRK, TYK2, UL97, VEGFR, Yes,
Zap70, Aurora-A, GSK3-alpha, HIPK1, HIPK2, HIP3, IRAKI, JNK1, JNK2,
JNK3, TRKB, CAMKII, CK1, CK2, RAF, GSK3Beta, MAPK1, MKK4, MKK7,
MST2, NEK2, AAK1, PKCalpha, PKD, RIPK2 and ROCK-II. (I think we
should consider whether to include such an expansive list. May be
restrict to those that are identified in the expanded cell plate
assay?)
[0133] In a preferred embodiment, the protein kinase is selected
from the group consisting of CDK1, CDK2, CDK3, CDK4, CDK5, CDK6,
CDK7, CDK8 and CDK9 and any combination thereof, as well as any
other CDK, as well as any CDK not yet identified. In a particularly
preferred embodiment, the protein kinase is selected from the group
consisting of CDK1, CDK2 and CDK9. In a particularly preferred
embodiment, the protein kinase is selected from the group
consisting of CDK9.
[0134] In a particular embodiment, CDK combinations of interest
include CDK4 and CDK9; CDK1, CDK2 and CDK9; CDK9 and CDK7; CDK9 and
CDK1; CDK9 and CDK2; CDK4, CDK6 and CDK9; CDK1, CDK2, CDK3, CDK4,
CDK6 and CDK9.
[0135] In other embodiments, the compounds of the present invention
are used for the treatment of protein kinase-associated disorders.
As used herein, the term "protein kinase-associated disorder"
includes disorders and states (e.g., a disease state) that are
associated with the activity of a protein kinase, e.g., the CDKs,
e.g., CDK1, CDK2 and/or CDK9. Non-limiting examples of protein
kinase-associated disorders include abnormal cell proliferation
(including protein kinase-associated cancers), viral infections,
fungal infections, autoimmune diseases and neurodegenerative
disorders.
[0136] Non-limiting examples of protein-kinase associated disorders
include proliferative diseases, such as viral infections,
auto-immune diseases, fungal disease, cancer, psoriasis, vascular
smooth cell proliferation associated with atherosclerosis,
pulmonary fibrosis, arthritis glomerulonephritis, chronic
inflammation, neurodegenerative disorders, such as Alzheimer's
disease, and post-surgical stenosis and restenosis. Protein
kinase-associated diseases also include diseases related to
abnormal cell proliferation, including, but not limited to, cancers
of the breast, ovary, cervix, prostate, testis, esophagus, stomach,
skin, lung, bone, colon, pancreas, thyroid, biliary passages,
buccal cavity and pharynx (oral), lip, tongue, mouth, pharynx,
small intestine, colon-rectum, large intestine, rectum, brain and
central nervous system, glioblastoma, neuroblastoma,
keratoacanthoma, epidermoid carcinoma, large cell carcinoma,
adenocarcinoma, adenocarcinoma, adenoma, adenocarcinoma, follicular
carcinoma, undifferentiated carcinoma, papillary carcinoma,
seminoma, melanoma, sarcoma, bladder carcinoma, liver carcinoma,
kidney carcinoma, myeloid disorders, lymphoid disorders, Hodgkin's,
hairy cells, and leukemia.
[0137] Additional non-limiting examples of protein
kinase-associated cancers include carcinomas, hematopoietic tumors
of lymphoid lineage, hematopoietic tumors of myeloid lineage,
tumors of mesenchymal origin, tumors of the central and peripheral
nervous system, melanoma, seminoma, teratocarcinoma, osteosarcoma,
xenoderoma pigmentosum, keratoctanthoma, thyroid follicular cancer
and Kaposi's sarcoma.
[0138] Protein kinase-associated disorders include diseases
associated with apoptosis, including, but not limited to, cancer,
viral infections, autoimmune diseases and neurodegenerative
disorders.
[0139] Non-limiting examples of protein-kinase associated disorders
include viral infections in a patient in need thereof, wherein the
viral infections include, but are not limited to, HIV, human
papilloma virus, herpes virus, poxyirus, Epstein-Barr virus,
Sindbis virus and adenovirus.
[0140] Non-limiting examples of protein-kinase associated disorders
include tumor angiogenesis and metastasis. Non-limiting examples of
protein-kinase associated disorders also include vascular smooth
muscle proliferation associated with atherosclerosis, postsurgical
vascular stenosis and restenosis, and endometriosis.
[0141] Further non-limiting examples of protein-kinase associated
disorders include those associated with infectious agents,
including yeast, fungi, protozoan parasites such as Plasitiodium
falciparum, and DNA and RNA viruses.
[0142] In another embodiment, the compound of the present invention
is further characterized as a modulator of a combination of protein
kinases, e.g., the CDKs, e.g., CDK1, CDK2 and/or CDK9. In certain
embodiments, a compound of the present invention is used for
protein kinase-associated diseases, and/or as an inhibitor of any
one or more protein kinases. It is envisioned that a use can be a
treatment of inhibiting one or more isoforms of protein
kinases.
[0143] The compounds of the invention are inhibitors of
cyclin-dependent kinase enzymes. Without being bound by theory,
inhibition of the CDK4/cyclin D1 complex blocks phosphorylation of
the Rb/inactive E2F complex, thereby preventing release of
activated E2F and ultimately blocking E2F-dependent DNA
transcription. This has the effect of inducing G.sub.1 cell cycle
arrest. In particular, the CDK4 pathway has been shown to have
tumor-specific deregulation and cytotoxic effects. Accordingly, the
ability to inhibit the activity of combinations of CDKs will be of
beneficial therapeutic use.
[0144] Furthermore, the cell's ability to respond and survive
chemotherapeutic assault may depend on rapid changes in
transcription or on activation of pathways which are highly
sensitive to CDK9/cyclinT1 (PTEF-b) activity. CDK9 inhibition may
sensitize cells to TNFalpha or TRAIL stimulation by inhibition of
NF-kB, or may block growth of cells by reducing myc-dependent gene
expression. CDK9 inhibition may also sensitize cells to genotoxic
chemotherapies, HDAC inhibition, or other signal transduction based
therapies.
[0145] As such, the compounds of the invention can lead to
depletion of anti-apoptotic proteins, which can directly induce
apoptosis or sensitize to other apoptotic stimuli, such as cell
cycle inhibition, DNA or microtubule damage or signal transduction
inhibition. Depletion of anti-apoptotic proteins by the compounds
of the invention may directly induce apoptosis or sensitize to
other apoptotic stimuli, such as cell cycle inhibition, DNA or
microtubule damage or signal transduction inhibition.
[0146] The compounds of the invention can be effective in
combination with chemotherapy, DNA damage arresting agents, or
other cell cycle arresting agents. The compounds of the invention
can also be effective for use in chemotherapy-resistant cells.
[0147] The present invention includes treatment of one or more
symptoms of cancer, inflammation, cardiac hypertrophy, and HIV
infection, as well as protein kinase-associated disorders as
described above, but the invention is not intended to be limited to
the manner by which the compound performs its intended function of
treatment of a disease. The present invention includes treatment of
diseases described herein in any manner that allows treatment to
occur, e.g., cancer, inflammation, cardiac hypertrophy, and HIV
infection.
[0148] In certain embodiments, the invention provides a
pharmaceutical composition of any of the compounds of the present
invention. In a related embodiment, the invention provides a
pharmaceutical composition of any of the compounds of the present
invention and a pharmaceutically acceptable carrier or excipient of
any of these compounds. In certain embodiments, the invention
includes the compounds as novel chemical entities.
[0149] In one embodiment, the invention includes a packaged protein
kinase-associated disorder treatment. The packaged treatment
includes a compound of the invention packaged with instructions for
using an effective amount of the compound of the invention for an
intended use.
[0150] The compounds of the present invention are suitable as
active agents in pharmaceutical compositions that are efficacious
particularly for treating protein kinase-associated disorders,
e.g., cancer, inflammation, cardiac hypertrophy, and HIV infection.
The pharmaceutical composition in various embodiments has a
pharmaceutically effective amount of the present active agent along
with other pharmaceutically acceptable excipients, carriers,
fillers, diluents and the like. The phrase, "pharmaceutically
effective amount" as used herein indicates an amount necessary to
administer to a host, or to a cell, issue, or organ of a host, to
achieve a therapeutic result, especially the regulating,
modulating, or inhibiting protein kinase activity, e.g., inhibition
of the activity of a protein kinase, or treatment of cancer,
inflammation, cardiac hypertrophy, and HIV infection.
[0151] In other embodiments, the present invention provides a
method for inhibiting the activity of a protein kinase. The method
includes contacting a cell with any of the compounds of the present
invention. In a related embodiment, the method further provides
that the compound is present in an amount effective to selectively
inhibit the activity of a protein kinase.
[0152] In other embodiments, the present invention provides a use
of any of the compounds of the invention for manufacture of a
medicament to treat cancer, inflammation, cardiac hypertrophy, and
HIV infection in a subject.
[0153] In other embodiments, the invention provides a method of
manufacture of a medicament, including formulating any of the
compounds of the present invention for treatment of a subject.
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS
[0154] The term "treat," "treated." "treating" or "treatment"
includes the diminishment or alleviation of at least one symptom
associated or caused by the state, disorder or disease being
treated. In certain embodiments, the treatment comprises the
induction of a protein kinase-associated disorder, followed by the
activation of the compound of the invention, which would in turn
diminish or alleviate at least one symptom associated or caused by
the protein kinase-associated disorder being treated. For example,
treatment can be diminishment of one or several symptoms of a
disorder or complete eradication of a disorder.
[0155] The term "use" includes any one or more of the following
embodiments of the invention, respectively: the use in the
treatment of protein kinase-associated disorders; the use for the
manufacture of pharmaceutical compositions for use in the treatment
of these diseases, e.g., in the manufacture of a medicament;
methods of use of compounds of the invention in the treatment of
these diseases; pharmaceutical preparations having compounds of the
invention for the treatment of these diseases; and compounds of the
invention for use in the treatment of these diseases; as
appropriate and expedient, if not stated otherwise. In particular,
diseases to be treated and are thus preferred for use of a compound
of the present invention are selected from cancer, inflammation,
cardiac hypertrophy, and HIV infection, as well as those diseases
that depend on the activity of protein kinases. The term "use"
further includes embodiments of compositions herein which bind to a
protein kinase sufficiently to serve as tracers or labels, so that
when coupled to a fluor or tag, or made radioactive, can be used as
a research reagent or as a diagnostic or an imaging agent.
[0156] The term "subject" is intended to include organisms, e.g.,
prokaryotes and eukaryotes, which are capable of suffering from or
afflicted with a disease, disorder or condition associated with the
activity of a protein kinase. Examples of subjects include mammals,
e.g., humans, dogs, cows, horses, pigs, sheep, goats, cats, mice,
rabbits, rats, and transgenic non-human animals. In certain
embodiments, the subject is a human, e.g., a human suffering from,
at risk of suffering from, or potentially capable of suffering from
cancer, inflammation, cardiac hypertrophy, and HIV infection, and
other diseases or conditions described herein (e.g., a protein
kinase-associated disorder). In another embodiment, the subject is
a cell.
[0157] The language "protein kinase-modulating compound,"
"modulator of protein kinase" or "protein kinase inhibitor" refers
to compounds that modulate, e.g., inhibit, or otherwise alter, the
activity of a protein kinase. Examples of protein kinase-modulating
compounds include compounds of the invention, i.e., Formula I and
Formula II, as well as the compounds of Table A, Table B, and Table
C (including pharmaceutically acceptable salts thereof, as well as
enantiomers, stereoisomers, rotamers, tautomers, diastereomers,
atropisomers or racemates thereof).
[0158] Additionally, a method of the invention includes
administering to a subject an effective amount of a protein
kinase-modulating compound of the invention, e.g., protein
kinase-modulating compounds of Formula I and Formula II, as well as
Table A, Table B, and Table C (including pharmaceutically
acceptable salts thereof, as well as enantiomers, stereoisomers,
rotamers, tautomers, diastereomers, atropisomers or racemates
thereof).
[0159] Where linking groups are specified by their conventional
chemical formula herein, written from left to right, they equally
encompass the chemically identical substituents that would result
from writing the structure from right to left, e.g., --CH.sub.2O--
is intended to include --OCH.sub.2-- for this purpose only.
[0160] The term "alkyl," by itself or as part of another
substituent, means, unless otherwise stated, a fully saturated
straight-chain (linear; unbranched) or branched chain, or a
combination thereof, having the number of carbon atoms specified,
if designated (i.e. C.sub.1-C.sub.10 means one to ten carbons).
Examples include, but are not limited to, groups such as methyl,
ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl,
homologs and isomers of, for example, n-pentyl, n-hexyl, n-heptyl,
n-octyl, and the like. If no size is designated, the alkyl groups
mentioned herein contain 1-10 carbon atoms, typically 1-8 carbon
atoms, and often 1-6 or 1-4 carbon atoms, and preferably 1-2 carbon
atoms. If the alkyl group is a branched alkyl group, and the number
of carbon atoms is not mentioned, the branched alkyl group will
consist of 3-8 carbon atoms, typically about 3-6 carbon atoms, and
particularly 3-4 carbon atoms.
[0161] The term "alkenyl" refers to unsaturated aliphatic groups
including straight-chain (linear; unbranched), branched-chain
groups, and combinations thereof, having the number of carbon atoms
specified, if designated, which contain at least one double bond
(--C.dbd.C--). All double bonds may be independently either (E) or
(Z) geometry, as well as mixtures thereof.
[0162] Examples of alkenyl groups include, but are not limited to,
--CH.sub.2--CH.dbd.CH--CH.sub.3; --CH.dbd.CH--CH.dbd.CH.sub.2 and
--CH.sub.2--CH.dbd.CH--CH(CH.sub.3)--CH.sub.2--CH.sub.3. If no size
is specified, the alkenyl groups discussed herein contain 2-6
carbon atoms.
[0163] The term "alkynyl" refers to unsaturated aliphatic groups
including straight-chain (linear; unbranched), branched-chain
groups, and combinations thereof, having the number of carbon atoms
specified, if designated, which contain at least one carbon-carbon
triple bond (--C.ident.C--). Examples of alkynyl groups include,
but are not limited to, --CH.sub.2--C.ident.C.ident.CH.sub.3;
--C.ident.C--C.ident.CH and
--CH.sub.2--C.ident.C--CH(CH.sub.3)--CH.sub.2--CH.sub.3. If no size
is specified, the alkynyl groups discussed herein contain 2-6
carbon atoms.
[0164] Alkynyl and alkenyl groups can contain more than one
unsaturated bond, or a mixture of double and triple bonds, and can
be otherwise substituted as described for alkyl groups.
[0165] The terms "alkoxy," "alkenyloxy." and "alkynyloxy" refer to
--O-alkyl, --O-alkenyl, and --O-alkynyl, respectively.
[0166] The term "cycloalkyl" by itself or in combination with other
terms, represents, unless otherwise stated, cyclic versions of
alkyl, alkenyl, or alkynyl, or mixtures thereof. Additionally,
cycloalkyl may contain fused rings, but excludes fused aryl and
heteroaryl groups, and cycloalkyl groups can be substituted unless
specifically described as unsubstituted. Examples of cycloalkyl
include, but are not limited to, cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl,
cyclohexynyl, cyclohexynyl, cyclohexadienyl, cyclopentadienyl,
cyclopentenyl, cycloheptyl, norbornyl, and the like. If no ring
size is specified, the cycloalkyl groups described herein contain
3-8 ring members, or 3-6 ring members.
[0167] The term "heterocyclic" or "heterocycloaklyl" or
"heterocyclyl," by itself or in combination with other terms,
represents a cycloalkyl radical containing at least one annular
carbon atom and at least one annular heteroatom selected from the
group consisting of O, N, P, Si and S, preferably from N, O and S,
wherein the ring is not aromatic but can contain unsaturations. The
nitrogen and sulfur atoms in a heterocyclic group may optionally be
oxidized and the nitrogen heteroatom may optionally be quaternized.
In many embodiments, the annular heteroatoms are selected from N, O
and S. The heterocyclic groups discussed herein, if not otherwise
specified, contain 3-10 ring members, and at least one ring member
is a heteroatom selected from N, O and S; commonly not more than
three of these heteroatoms are included in a heterocyclic group,
and generally not more than two of these heteroatoms are present in
a single ring of the heterocyclic group. The heterocyclic group can
be fused to an additional carboclic, heterocyclic, or aryl ring. A
heterocyclic group can be attached to the remainder of the molecule
at an annular carbon or annular heteroatom, and the heterocyclic
groups can be substituted as described for alkyl groups.
Additionally, heterocyclic may contain fused rings, but excludes
fused systems containing a heteroaryl group as part of the fused
ring system. Examples of heterocyclic groups include, but are not
limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl,
2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl,
tetrahydrofuran-2-yl, 1,2,3,4-tetrahydropyridyl, dihydroindole
(indoline), tetrahydrofuran-3-yl, tetrahydrothien-2-yl,
tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the
like.
[0168] As with other moieties described herein, heterocycloalkyl
moieties can be unsubstituted, or substituted with various
substituents known in the art, e.g., hydroxy, halo, oxo (C.dbd.O),
alkylimino (RN.dbd., wherein R is a loweralkyl or loweralkoxy
group), amino, alkylamino, dialkylamino, acylaminoalkyl, alkoxy,
thioalkoxy, polyalkoxy, loweralkyl, cycloalkyl or haloalkyl.
Non-limiting examples of substituted heterocycloalkyl groups
include the following, where each moiety may be attached to the
parent molecule at any available valence:
##STR00006##
[0169] Also included within heterocyclic are piperidine,
morpholine, thiomorpholine, piperazine, pyrrolidine,
tetrahydrofuran, oxetane, oxepane, oxirane, tetrahydrothiofuran,
thiepane, thiirane, and optionally substituted versions of each of
these.
[0170] The terms "cycloalkyloxy" and "hetcrocycloalkyloxy" refer to
--O-cycloalkyl and --O-heterocycloalkyl groups, respectively (e.g.,
cyclopropoxy, 2-piperidinyloxy, and the like).
[0171] The term "aryl" means, unless otherwise stated, an aromatic
hydrocarbon group which can be a single ring or multiple rings
(e.g., from 1 to 3 rings) which are fused together. Aryl may
contain fused rings, wherein one or more of the rings is optionally
cycloalkyl, but not including heterocyclic or heteroaromatic rings;
a fused system containing at least one heteroaromatic ring is
described as a heteroaryl group, and a phenyl ring fused to a
heterocyclic ring is described herein as a heterocyclic group. An
aryl group will include a fused ring system wherein a phenyl ring
is fused to a cycloalkyl ring. Examples of aryl groups include, but
are not limited to, phenyl, 1-naphthyl, tetrahydro-naphthalene,
dihydro-1H-indene, 2-naphthyl, tetrahydronaphthyl and the like.
[0172] The term "heteroaryl" as used herein refers to groups
comprising a single ring or two or three fused rings, where at
least one of the rings is an aromatic ring that contain from one to
four heteroatoms selected from N, O, and S as ring members (i.e.,
it contains at least one heteroaromatic ring), wherein the nitrogen
and sulfur atoms are optionally oxidized, and the nitrogen atom(s)
are optionally quaternized. A heteroaryl group can be attached to
the remainder of the molecule through an annular carbon or annular
heteroatom, and it can be attached through any ring of the
heteroaryl moiety, if that moiety is bicyclic or tricyclic.
Heteroaryl may contain fused rings, wherein one or more of the
rings is optionally cycloalkyl or heterocycloalkyl or aryl,
provided at least one of the rings is a heteroaromatic ring.
Non-limiting examples of heteroaryl groups are 1-pyrrolyl,
2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl,
pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl,
3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl,
5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl,
3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl,
purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,
2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl.
Substituents for each of the above noted aryl and heteroaryl ring
systems are selected from the group of acceptable substituents
described below.
[0173] Aryl and/or heteroaryl groups commonly contain up to four
substituents per ring (0-4), and sometimes contain 0-3 or 0-2
substituents. The terms "aryloxy" and "heteroaryloxy" refer to aryl
and heteroaryl groups, respectively, attached to the remainder of
the molecule via an oxygen linker (--O--).
[0174] The term "arylalkyl" or "aralkyl" designates an alkyl-linked
aryl group, where the alkyl portion is attached to the parent
structure and the aryl is attached to the alkyl portion of the
arylalkyl moiety. Examples are benzyl, phenethyl, and the like.
"Heteroarylalkyl" or "heteroaralkyl" designates a heteroaryl moiety
attached to the parent structure via an alkyl residue. Examples
include furanylmethyl, pyridinylmethyl, pyrimidinylethyl, and the
like. Aralkyl and heteroaralkyl also include substituents in which
at least one carbon atom of the alkyl group is present in the alkyl
group and wherein another carbon of the alkyl group has been
replaced by, for example, an oxygen atom (e.g., phenoxymethyl,
2-pyridylmethoxy, 3-(1-naphthyloxy)propyl, and the like).
[0175] The terms "halo" or "halogen," by themselves or as part of
another substituent, mean, unless otherwise stated, a fluorine,
chlorine, bromine, or iodine atom. Additionally, terms such as
"haloalkyl," are meant to include monohaloalkyl and perhaloalkyl.
For example, the term "halo(C.sub.1-C.sub.4)alkyl" is meant to
include, but not be limited to, trifluoromethyl,
2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.
The prefix "perhalo" refers to the respective group wherein all
available valences are replaced by halo groups. For example
"perhaloalkyl" includes --CCl.sub.3, --CF.sub.3,
--CCl.sub.2CF.sub.3, and the like. The terms "perfluoroalkyl" and
"perchloroalkyl" are a subsets of perhaloalkyl wherein all
available valences are replaced by fluoro and chloro groups,
respectively. Non limiting examples of perfluoroalkyl include
--CF.sub.3 and --CF.sub.2CF.sub.3. Non limiting examples of
perchloroalkyl include --CCl.sub.3 and --CCl.sub.2CCl.sub.3.
[0176] "Amino" refers herein to the group --NH.sub.2 or --NRR',
where R and R' are each independently selected from hydrogen or an
alkyl (e.g, lower alkyl). The term "arylamino" refers herein to the
group --NRR' where R is aryl and R' is hydrogen, alkyl, or an aryl.
The term "aralkylamino" refers herein to the group --NRR' where R
is an aralkyl and R' is hydrogen, an alkyl, an aryl, or an aralkyl.
"Substituted amino" refers to an amino wherein at least one of R
and R' is not H, i.e., the amino has at least one substituent group
on it. The term alkylamino refers to -alkyl-NRR' where R and R' are
each independently selected from hydrogen or an alkyl (e.g, lower
alkyl).
[0177] The term "aminocarbonyl" refers herein to the group
--C(O)--NH.sub.2, i.e., it is attached to the base structure
through the carbonyl carbon atom. "Substituted aminocarbonyl"
refers herein to the group --C(O)--NRR' where R is alkyl and R' is
hydrogen or an alkyl. The term "arylaminocarbonyl" refers herein to
the group --C(O)--NRR' where R is an aryl and R' is hydrogen, alkyl
or aryl. "Aralkylaminocarbonyl" refers herein to the group
--C(O)--NRR' where R is aralkyl and R' is hydrogen, alkyl, aryl, or
aralkyl.
[0178] "Aminosulfonyl" refers herein to the group
--S(O).sub.2--NH.sub.2. "Substituted aminosulfonyl" refers herein
to the group --S(O).sub.2--NRR' where R is alkyl and R' is hydrogen
or an alkyl. The term "aralkylaminosulfonlyaryl" refers herein to
the group -aryl-S(O).sub.2--NH-aralkyl.
[0179] "Carbonyl" refers to the divalent group --C(O)--.
[0180] The term "sulfonyl" refers herein to the group --SO.sub.2--.
"Alkylsulfonyl" refers to a substituted sulfonyl of the structure
--SO.sub.2R in which R is alkyl. Alkylsulfonyl groups employed in
compounds of the present invention are typically loweralkylsulfonyl
groups having from 1 to 6 carbon atoms in R. Thus, exemplary
alkylsulfonyl groups employed in compounds of the present invention
include, for example, methylsulfonyl (i.e., where R is methyl),
ethylsulfonyl (i.e., where R is ethyl), propylsulfonyl (i.e., where
R is propyl), and the like. The term "arylsulfonyl" refers herein
to the group --SO.sub.2-aryl. The term "aralkylsulfonyl" refers
herein to the group --SO.sub.2-aralkyl. The term "sulfonamido"
refers herein to --SO.sub.2NH.sub.2, or to --SO.sub.2NRR' if
substituted.
[0181] Unless otherwise stated, each radical/moiety described
herein (e.g., "alkyl," "cycloalkyl," "heterocycloalkyl," "aryl,"
"heteroaryl," "alkoxy," etc.) is meant to include both substituted
and unsubstituted forms.
[0182] "Optionally substituted" as used herein indicates that the
particular group or groups being described may have no non-hydrogen
substituents (i.e., it can be unsubstituted), or the group or
groups may have one or more non-hydrogen substituents. If not
otherwise specified, the total number of such substituents that may
be present is equal to the number of H atoms present on the
unsubstituted form of the group being described. Typically, a group
will contain up to three (0-3) substituents. Where an optional
substituent is attached via a double bond, such as a carbonyl
oxygen (.dbd.O), the group takes up two available valences on the
group being substituted, so the total number of substituents that
may be included is reduced according to the number of available
valences. Suitable substituent groups include, for example,
hydroxyl, nitro, amino, imino, cyano, halo, thio, sulfonyl,
thioamido, amidino, imidino, oxo, oxamidino, methoxamidino,
imidino, guanidino, sulfonamido, carboxyl, formyl, loweralkyl,
loweralkoxy, loweralkoxyalkyl, alkylcarbonyl, aminocarbonyl,
arylcarbonyl, aralkylcarbonyl, carbonylamino, heteroarylcarbonyl,
heteroaralkylcarbonyl, alkylthio, aminoalkyl, cyanoalkyl, aryl,
alkylamino, alkylsulfonyl, aralkylamino, alkylcarbonylamino,
carbonyl, piperidinyl, morpholinyl, pyrrolidinyl and the like.
Deuterium, when introduced into a compound at levels at least
5.times. above natural abundance, can also be considered a
substituent for purposes of describing the compounds herein. Note
that because deuterium is an isotope of hydrogen that does not
substantially change the shape of the molecule, deuterium is exempt
from the typical numerical limitations placed on numbers of
substituents: deuterium (D) can be included in place of hydrogen
(H) in addition to other substituents and should not be counted in
the numerical limitations that apply to other substituents.
[0183] A substituent group can itself be substituted by the same
groups described herein for the corresponding type of structure.
The group substituted onto the substituted group can be carboxyl,
halo, nitro, amino, cyano, hydroxyl, loweralkyl, loweralkenyl,
loweralkynyl, loweralkoxy, aminocarbonyl, --SR, thioamido,
--SO.sub.3H, --SO.sub.2R, N-methylpyrrolidinyl, piperidinyl,
piperazinyl, N-methylpiperazinyl, 4-chloropyrimidinyl, pyrindinyl,
tetrahydropyranyl (or heterocycloalkyl, heteroaryl?) or cycloalkyl,
where R is typically hydrogen or loweralkyl.
[0184] When the substituted substituent includes a straight chain
group, the substituent can occur either within the chain (e.g.,
2-hydroxypropyl, 2-aminobutyl, and the like) or at the chain
terminus (e.g., 2-hydroxyethyl, 3-cyanopropyl, and the like).
Substituted substituents can be straight chain, branched or cyclic
arrangements of covalently bonded carbon or heteroatoms (N, O or
S).
[0185] The term "cycloalkyl" may be used herein to describe a
carbocyclic non-aromatic group that is connected via a ring carbon
atom, and "cycloalkylalkyl" may be used to describe a carbocyclic
non-aromatic group that is connected to the molecule through an
alkyl linker. Similarly, "heterocyclyl" may be used to describe a
non-aromatic cyclic group that contains at least one heteroatom as
a ring member and that is connected to the molecule via a ring
atom, which may be C or N; and "heterocyclylalkyl" may be used to
describe such a group that is connected to another molecule through
a linker. The sizes and substituents that are suitable for the
cycloalkyl, cycloalkylalkyl, heterocyclyl, and heterocyclylalkyl
groups are the same as those described above for alkyl groups. As
used herein, these terms also include rings that contain a double
bond or two, as long as the ring is not aromatic.
[0186] As used herein, "isomer" includes all stereoisomers of the
compounds referred to in the formulas herein, including
enantiomers, diastereomers, as well as all conformers, rotamers,
and tautomers, unless otherwise indicated. The invention includes
all enantiomers of any chiral compound disclosed, in either
substantially pure levorotatory or dextrorotatory form, or in a
racemic mixture, or in any ratio of enantiomers. For compounds
disclosed as an (R)-enantiomer, the invention also includes the
(S)-enantiomer, for compounds disclosed as the (S)-enantiomer, the
invention also includes the (R)-enantiomer. The invention includes
any diastereomers of the compounds referred to in the above
formulas in diastereomerically pure form and in the form of
mixtures in all ratios.
[0187] Unless stereochemistry is explicitly indicated in a chemical
structure or chemical name, the chemical structure or chemical name
is intended to embrace all possible stereoisomers, conformers,
rotamers, and tautomers of the compound depicted. For example, a
compound containing a chiral carbon atom is intended to embrace
both the (R) enantiomer and the (S) enantiomer, as well as mixtures
of enantiomers, including racemic mixtures; and a compound
containing two chiral carbons is intended to embrace all
enantiomers and diastereomers (including (R,R), (S,S), (R,S), and
(R,S) isomers).
[0188] In all uses of the compounds of the formulas disclosed
herein, the invention also includes use of any or all of the
stereochemical, enantiomeric, diastereomeric, conformational,
rotomeric, tautomeric, solvate, hydrate, polymorphic, crystalline
form, non-crystalline form, salt, pharmaceutically acceptable salt,
metabolite and prodrug variations of the compounds as
described.
[0189] The term "heteroatom" includes atoms of any element other
than carbon or hydrogen. Preferred heteroatoms are nitrogen,
oxygen, sulfur and phosphorus.
[0190] Additionally, the phrase "any combination thereof" implies
that any number of the listed functional groups and molecules may
be combined to create a larger molecular architecture. For example,
the terms "phenyl," "carbonyl" (or ".dbd.O"), "--O--," "--OH," and
C.sub.1-6 (i.e., --CH.sub.3 and --CH.sub.2CH.sub.2CH.sub.2--) can
be combined to form a 3-methoxy-4-propoxybenzoic acid substituent.
It is to be understood that when combining functional groups and
molecules to create a larger molecular architecture, hydrogens can
be removed or added, as required to satisfy the valence of each
atom.
[0191] The description of the disclosure herein should be construed
in congruity with the laws and principals of chemical bonding. For
example, it may be necessary to remove a hydrogen atom in order
accommodate a substitutent at any given location. Furthermore, it
is to be understood that definitions of the variables (i.e., "R
groups"), as well as the bond locations of the generic formulae of
the invention (e.g., formulas I or II), will be consistent with the
laws of chemical bonding known in the art. It is also to be
understood that all of the compounds of the invention described
above will further include bonds between adjacent atoms and/or
hydrogens as required to satisfy the valence of each atom. That is,
bonds and/or hydrogen atoms are added to provide the following
number of total bonds to each of the following types of atoms:
carbon: four bonds; nitrogen: three bonds; oxygen: two bonds; and
sulfur: two-six bonds.
[0192] As used herein, "isomer" includes all stereoisomers of the
compounds referred to in the formulas herein, including
enantiomers, diastereomers, as well as all conformers, rotamers,
and tautomers, unless otherwise indicated. The invention includes
all enantiomers of any chiral compound disclosed, in either
substantially pure levorotatory or dextrorotatory form, or in a
racemic mixture, or in any ratio of enantiomers. For compounds
disclosed as an (R)-enantiomer, the invention also includes the
(S)-enantiomer, for compounds disclosed as the (S)-enantiomer, the
invention also includes the (R)-enantiomer. The invention includes
any diastereomers of the compounds referred to in the above
formulas in diastereomerically pure form and in the form of
mixtures in all ratios.
[0193] Unless stereochemistry is explicitly indicated in a chemical
structure or chemical name, the chemical structure or chemical name
is intended to embrace all possible stereoisomers, conformers,
rotamers, and tautomers of the compound depicted. For example, a
compound containing a chiral carbon atom is intended to embrace
both the (R) enantiomer and the (S) enantiomer, as well as mixtures
of enantiomers, including racemic mixtures; and a compound
containing two chiral carbons is intended to embrace all
enantiomers and diastereomers (including (R,R), (S,S), (R,S), and
(R,S) isomers).
[0194] In all uses of the compounds of the formulas disclosed
herein, the invention also includes use of any or all of the
stereochemical, enantiomeric, diastereomeric, conformational,
rotomeric, tautomeric, solvate, hydrate, polymorphic, crystalline
form, non-crystalline form, salt, pharmaceutically acceptable salt,
metabolite and prodrug variations of the compounds as
described.
[0195] It will also be noted that the substituents of some of the
compounds of this invention include isomeric cyclic structures. It
is to be understood accordingly that constitutional isomers of
particular substituents are included within the scope of this
invention, unless indicated otherwise. For example, the term
"tetrazole" includes tetrazole, 2H-tetrazole, 3H-tetrazole,
4H-tetrazole and 5H-tetrazole.
[0196] Certain compounds of the present invention can exist in
unsolvated forms as well as solvated forms (i.e., solvates).
Compounds of the invention may also include hydrated forms (i.e.,
hydrates). In general, the solvated and hydrated forms are
equivalent to unsolvated forms for purposes of biological utility
and are encompassed within the scope of the present invention. The
invention also includes all polymorphs, including crystalline and
non-crystalline forms. In general, all physical forms are
equivalent for the uses contemplated by the present invention and
are intended to be within the scope of the present invention.
[0197] The present invention includes all salt forms of the
compounds described herein, as well as methods of using such salts.
The invention also includes all non-salt forms of any salt of a
compound named herein, as well as other salts of any salt of a
compound named herein. In one embodiment, the salts of the
compounds comprise pharmaceutically acceptable salts.
"Pharmaceutically acceptable salts" are those salts which retain
the biological activity of the free compounds and which can be
administered as drugs or pharmaceuticals to humans and/or animals.
The desired salt of a basic functional group of a compound may be
prepared by methods known to those of skill in the art by treating
the compound with an acid. Examples of inorganic acids include, but
are not limited to, hydrochloric acid, hydrobromic acid, sulfuric
acid, nitric acid, and phosphoric acid. Examples of organic acids
include, but are not limited to, formic acid, acetic acid,
propionic acid, glycolic acid, hippuric, pyruvic acid, oxalic acid,
maleic acid, malonic acid, succinic acid, fumaric acid, tartaric
acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,
sulfonic acids, and salicylic acid. The desired salt of an acidic
functional group of a compound can be prepared by methods known to
those of skill in the art by treating the compound with a base.
Examples of inorganic salts of acid compounds include, but are not
limited to, alkali metal and alkaline earth salts, such as sodium
salts, potassium salts, magnesium salts, and calcium salts;
ammonium salts; and aluminum salts. Examples of organic salts of
acid compounds include, but are not limited to, procaine,
dibenzylamine, N-ethylpiperidine, N,N'-dibenzylethylenediamine, and
triethylamine salts.
[0198] Pharmaceutically acceptable metabolites and prodrugs of the
compounds referred to in the formulas herein are also embraced by
the invention. The term "pharmaceutically acceptable prodrugs" as
used herein refers to those prodrugs of the compounds of the
present invention which are, within the scope of sound medical
judgment, suitable for use in contact with the tissues of humans
and lower animals without undue toxicity, irritation, allergic
response, and the like, commensurate with a reasonable benefit/risk
ratio, and effective for their intended use, as well as the
zwitterionic forms, where possible, of the compounds of the
invention. The term "prodrug" refers to compounds that are rapidly
transformed in vivo to yield the parent compound of the above
formula, for example by hydrolysis in blood. A thorough discussion
is provided in T. Higuchi and V. Stella, PRO-DRUGS AS NOVEL
DELIVERY SYSTEMS, Vol. 14 of the A.C.S. Symposium Series, and in
Edward B. Roche, ed., BIOREVERSIBLE CARRIERS IN DRUG DESIGN,
American Pharmaceutical Association and Pergamon Press, 1987.
[0199] Pharmaceutically acceptable esters of the compounds referred
to in the formulas herein are also embraced by the invention. As
used herein, the term "pharmaceutically acceptable ester" refers to
esters, which hydrolyze in vivo and include those that break down
readily in the human body to leave the parent compound or a salt
thereof. Suitable ester groups include, for example, those derived
from pharmaceutically acceptable aliphatic carboxylic acids,
particularly alkanoic, alkenoic, cycloalkanoic and alkanedioic
acids, in which each alkyl or alkenyl moiety advantageously has not
more than 6 carbon atoms. Examples of particular esters include
formates, acetates, propionates, butyrates, acrylates and
ethylsuccinates.
[0200] The invention further provides deuterated versions of the
above-described compounds. As used herein, "deuterated version"
refers to a compound in which at least one hydrogen atom is
enriched in the isotope deuterium beyond the natural rate of
deuterium occurrence. Typically, the hydrogen atom is enriched to
be at least 50% deuterium, frequently at least 75% deuterium, and
preferably at least about 90% deuterium. Optionally, more than one
hydrogen atom can be replaced by deuterium. For example, a methyl
group can be deuterated by replacement of one hydrogen with
deuterium (i.e., it can be --CH.sub.2D), or it can have all three
hydrogen atoms replaced with deuterium (i.e., it can be
--CD.sub.3). In each case, D signifies that at least 50% of the
corresponding H is present as deuterium.
[0201] A substantially pure compound means that the compound is
present with no more than 15% or no more than 10% or no more than
5% or no more than 3% or no more than 1% of the total amount of
compound as impurity and/or in a different form. For instance,
substantially pure S,S compound means that no more than 15% or no
more than 10% or no more than 5% or no more than 3% or no more than
1% of the total R,R; S,R; and R,S forms are present.
[0202] As used herein, "therapeutically effective amount" indicates
an amount that results in a desired pharmacological and/or
physiological effect for the condition. The effect may be
prophylactic in terms of completely or partially preventing a
condition or symptom thereof and/or may be therapeutic in terms of
a partial or complete cure for the condition and/or adverse effect
attributable to the condition. Therapeutically effective amounts of
the compounds of the invention generally include any amount
sufficient to detectably inhibit Raf activity by any of the assays
described herein, by other Raf kinase activity assays known to
those having ordinary skill in the art or by detecting an
inhibition or alleviation of symptoms of cancer.
[0203] As used herein, the term "pharmaceutically acceptable
carrier," and cognates thereof, refers to adjuvants, binders,
diluents, etc. known to the skilled artisan that are suitable for
administration to an individual (e.g., a mammal or non-mammal).
Combinations of two or more carriers are also contemplated in the
present invention. The pharmaceutically acceptable carrier(s) and
any additional components, as described herein, should be
compatible for use in the intended route of administration (e.g.,
oral, parenteral) for a particular dosage form. Such suitability
will be easily recognized by the skilled artisan, particularly in
view of the teaching provided herein. Pharmaceutical compositions
described herein include at least one pharmaceutically acceptable
carrier or excipient; preferably, such compositions include at
least one carrier or excipient other than or in addition to
water.
[0204] As used herein, the term "pharmaceutical agent" or
"additional pharmaceutical agent," and cognates of these terms, are
intended to refer to active agents other than the claimed compounds
of the invention, for example, drugs, which are administered to
elicit a therapeutic effect. The pharmaceutical agent(s) may be
directed to a therapeutic effect related to the condition that a
claimed compound is intended to treat or prevent (e.g., conditions
mediated by Raf kinase, including, but not limited to those
conditions described herein (e.g., cancer)) or, the pharmaceutical
agent may be intended to treat or prevent a symptom of the
underlying condition (e.g., tumor growth, hemorrhage, ulceration,
pain, enlarged lymph nodes, cough, jaundice, swelling, weight loss,
cachexia, sweating, anemia, paraneoplastic phenomena, thrombosis,
etc.) or to further reduce the appearance or severity of side
effects of administering a claimed compound.
[0205] When used with respect to methods of treatment/prevention
and the use of the compounds and formulations thereof described
herein, an individual "in need thereof" may be an individual who
has been diagnosed with or previously treated for the condition to
be treated. With respect to prevention, the individual in need
thereof may also be an individual who is at risk for a condition
(e.g., a family history of the condition, life-style factors
indicative of risk for the condition, etc.). Typically, when a step
of administering a compound of the invention is disclosed herein,
the invention further contemplates a step of identifying an
individual or subject in need of the particular treatment to be
administered or having the particular condition to be treated.
[0206] In some embodiments, the individual is a mammal, including,
but not limited to, bovine, horse, feline, rabbit, canine, rodent,
or primate. In some embodiments, the mammal is a primate. In some
embodiments, the primate is a human. In some embodiments, the
individual is human, including adults, children and premature
infants. In some embodiments, the individual is a non-mammal. In
some variations, the primate is a non-human primate such as
chimpanzees and other apes and monkey species. In some embodiments,
the mammal is a farm animal such as cattle, horses, sheep, goats,
and swine; pets such as rabbits, dogs, and cats; laboratory animals
including rodents, such as rats, mice, and guinea pigs; and the
like. Examples of non-mammals include, but are not limited to,
birds, and the like. The term "individual" does not denote a
particular age or sex.
[0207] In some variations, the individual has been identified as
having one or more of the conditions described herein.
Identification of the conditions as described herein by a skilled
physician is routine in the art (e.g., via blood tests, X-rays, CT
scans, endoscopy, biopsy, etc.) and may also be suspected by the
individual or others, for example, due to tumor growth, hemorrhage,
ulceration, pain, enlarged lymph nodes, cough, jaundice, swelling,
weight loss, cachexia, sweating, anemia, paraneoplastic phenomena,
thrombosis, etc. In some embodiments, the individual has further
been identified as having a cancer that expresses a mutated Raf,
such as a mutated B-Raf.
[0208] In some embodiments, the individual has been identified as
susceptible to one or more of the conditions as described herein.
The susceptibility of an individual may be based on any one or more
of a number of risk factors and/or diagnostic approaches
appreciated by the skilled artisan, including, but not limited to,
genetic profiling, family history, medical history (e.g.,
appearance of related conditions), lifestyle or habits.
[0209] As used herein and in the appended claims, the singular
forms "a", "an" and "the" include plural forms, unless the context
clearly dictates otherwise.
[0210] Unless defined otherwise or clearly indicated by context,
all technical and scientific terms used herein have the same
meaning as commonly understood by one of ordinary skill in the art
to which this invention belongs.
General Synthetic Methods
[0211] The compounds disclosed herein can be prepared from readily
available starting materials using the following general methods
and procedures. It will be appreciated that where typical or
preferred process conditions (i.e., reaction temperatures, times,
mole ratios of reactants, solvents, pressures, etc.) are given,
other process conditions can also be used unless otherwise stated.
Optimum reaction conditions may vary with the particular reactants
or solvent used, but such conditions can be determined by one
skilled in the art by routine optimization procedures.
[0212] Additionally, as will be apparent to those skilled in the
art, conventional protecting groups may be necessary to prevent
certain functional groups from undergoing undesired reactions.
Suitable protecting groups for various functional groups as well as
suitable conditions for protecting and deprotecting particular
functional groups are well known in the art. For example, numerous
protecting groups are described in T. W. Greene and G. M. Wuts,
Protecting Groups in Organic Synthesis, Third Edition, Wiley, New
York, 1999, and references cited therein.
[0213] Furthermore, the compounds disclosed herein may contain one
or more chiral centers. Accordingly, if desired, such compounds can
be prepared or isolated as pure stereoisomers, i.e., as individual
enantiomers or diastereomers, or as stereoisomer enriched mixtures.
All such stereoisomers (and enriched mixtures) are included within
the scope of the embodiments, unless otherwise indicated. Pure
stereoisomers (or enriched mixtures) may be prepared using, for
example, optically active starting materials or stereoselective
reagents well-known in the art. Alternatively, racemic mixtures of
such compounds can be separated using, for example, chiral column
chromatography, chiral resolving agents and the like.
[0214] The starting materials for the following reactions are
generally known compounds or can be prepared by known procedures or
obvious modifications thereof. For example, many of the starting
materials are available from commercial suppliers such as Aldrich
Chemical Co. (Milwaukee, Wis., USA), Bachem (Torrance, Calif.,
USA), Emka-Chemce or Sigma (St. Louis, Mo., USA). Others may be
prepared by procedures, or obvious modifications thereof, described
in standard reference texts such as Fieser and Fieser's Reagents
for Organic Synthesis, Volumes 1-15 (John Wiley and Sons, 1991),
Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals
(Elsevier Science Publishers, 1989), Organic Reactions, Volumes
1-40 (John Wiley and Sons, 1991), March's Advanced Organic
Chemistry, (John Wiley and Sons, 4.sup.th Edition), and Larock's
Comprehensive Organic Transformations (VCH Publishers Inc.,
1989).
[0215] The various starting materials, intermediates, and compounds
of the embodiments may be isolated and purified where appropriate
using conventional techniques such as precipitation, filtration,
crystallization, evaporation, distillation, and chromatography.
Characterization of these compounds may be performed using
conventional methods such as by melting point, mass spectrum,
nuclear magnetic resonance, and various other spectroscopic
analyses.
[0216] Compounds of the embodiments may generally be prepared using
a number of methods familiar to one of skill in the art, and may
generally be made in accordance with the following reaction Schemes
1 and 2, which are described in detail in the Examples below.
EXAMPLES
[0217] Referring to the examples that follow, compounds of the
embodiments were synthesized using the methods described herein, or
other methods known to one skilled in the art.
[0218] The compounds and/or intermediates were characterized by
high performance liquid chromatography (HPLC) using a Waters
Millenium chromatography system with a 2695 Separation Module
(Milford, Mass.). The analytical columns were reversed phase
Phenomenex Luna C18 5.mu., 4.6.times.50 mm, from Alltech
(Deerfield, Ill.). A gradient elution was used (flow 2.5 mL/min),
typically starting with 5% acetonitrile/95% water and progressing
to 100% acetonitrile over a period of 10 minutes. All solvents
contained 0.1% trifluoroacetic acid (TFA). Compounds were detected
by ultraviolet light (UV) absorption at either 220 or 254 nm. HPLC
solvents were from Burdick and Jackson (Muskegan, Mich.), or Fisher
Scientific (Pittsburgh, Pa.).
[0219] In some instances, purity was assessed by thin layer
chromatography (TLC) using glass or plastic backed silica gel
plates, such as, for example, Baker-Flex Silica Gel 1B2-F flexible
sheets. TLC results were readily detected visually under
ultraviolet light, or by employing well known iodine vapor and
other various staining techniques.
[0220] Mass spectrometric analysis was performed on LCMS
instruments: Waters System (Acuity UPLC and a Micromass ZQ mass
spectrometer, Column: Acuity HSS C18 1.8-micron, 2.1.times.50 mm;
gradient: 5-95% acetonitrile in water with 0.05% TFA over a 1.8 min
period; flow rate 1.2 mL/min; molecular weight range 200-1500; cone
Voltage 20 V; column temperature 50.degree. C.). All masses were
reported as those of the protonated parent ions.
[0221] GCMS analysis is performed on a Hewlett Packard instrument
(HP6890 Series gas chromatograph with a Mass Selective Detector
5973; injector volume: 1 L; initial column temperature: 50.degree.
C.; final column temperature: 250.degree. C.; ramp time: 20
minutes; gas flow rate: 1 mL/min; column: 5% phenyl methyl
siloxane, Model No. HP 190915-443, dimensions: 30.0 m.times.25
m.times.0.25 m).
[0222] Nuclear magnetic resonance (NMR) analysis was performed on
some of the compounds with a Varian 300 MHz NMR (Palo Alto, Calif.)
or Varian 400 MHz MR NMR (Palo Alto, Calif.). The spectral
reference was either TMS or the known chemical shift of the
solvent. Some compound samples were run at elevated temperatures
(e.g., 75.degree. C.) to promote increased sample solubility.
[0223] The purity of some of the compounds is assessed by elemental
analysis (Desert Analytics, Tucson, Ariz.).
[0224] Melting points are determined on a Laboratory Devices
Mel-Temp apparatus (Holliston, Mass.).
[0225] Preparative separations are carried out using a Combiflash
Rf system (Teledyne Isco, Lincoln, Nebr.) with RediSep silica gel
cartridges (Teledyne Isco, Lincoln, Nebr.) or SiliaSep silica gel
cartridges (Silicycle Inc., Quebec City, Canada) or by flash column
chromatography using silica gel (230-400 mesh) packing material, or
by HPLC using a Waters 2767 Sample Manager, C-18 reversed phase
column, 30.times.50 mm, flow 75 mL/min. Typical solvents employed
for the Combiflash Rf system and flash column chromatography are
dichloromethane, methanol, ethyl acetate, hexane, heptane, acetone,
aqueous ammonia (or ammonium hydroxide), and triethyl amine.
Typical solvents employed for the reverse phase HPLC are varying
concentrations of acetonitrile and water with 0.1% trifluoroacetic
acid.
[0226] The examples below as well as throughout the application,
the following abbreviations have the following meanings. If not
defined, the terms have their generally accepted meanings.
ABBREVIATIONS
ACN: Acetonitrile
[0227] BINAP: 2,2'-bis(diphenylphosphino)-1,1'-binapthyl
DCM: Dichloromethane
[0228] DIEA: diisopropylethylamine
DIPEA: N,N-diisopropylethylamine
[0229] DME: 1,2-dimethoxyethane
DMF: N,N-dimethylformamide
[0230] DMSO dimethyl sulfoxide DPPF
1,1'-bis(diphenylphosphino)ferrocene eq equivalent EtOAc ethyl
acetate EtOH ethanol HATU
2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate HPLC high performance liquid chromatography
MCPBA meta-chloroperoxybenzoic acid MeOH methanol
NBS N-bromosuccinimide
[0231] NMP N-methyl-2-pyrrolidone Rt retention time THF
tetrahydrofuran
Synthetic Examples
[0232] Compounds of the present invention can be synthesized by the
schemes outlined b
##STR00007## ##STR00008##
[0233] As shown in Scheme 1a, synthesis can start with a
functionalized pyridine or pyrimidine I wherein LG is a leaving
group such as F, Cl, OTf, and the like. X can be a functional group
like Cl, Br, I or OTf. Compound I can be converted into boronic
acid or boronic ester II by:
[0234] 1) PdCl.sub.2(dppf) DCM adduct, potassium acetate,
bis(pinacolato)diboron heating from 30-120.degree. C. in solvents
such as THF, DMF, DME, DMA, toluene and dioxane; and 2) In a
solvent such as THF or diethylether, anion halogen exchange by
addition of nBuLi or LDA followed by quenching the anion with
triisopropyl borate. Upon hydrolysis a boronic acid can be
obtained. Suzuki cross-coupling reaction between compound II and
pyridine or pyrazine III then gives bi-heteroaryl intermediate IV.
The SN.sub.AR reaction between IV and a functionalized amine
NH.sub.2R.sub.1' under basic condition (DIEA, TEA, lutidine,
pyridine) in a solvent such as DMF, THF, DMSO, NMP, dioxane with
heating (30-130.degree. C.) can give compound V. When R.sub.1' is
not identical to R.sub.1, further functional manipulation is needed
to obtain VI. When R.sub.1' is identical to R.sub.1, compound V
will be the same as compound VI. Alternatively, VI can be obtained
by following Scheme 1b. In which the Suzuki cross-coupling step is
carried out between I and VII. Boronic acid or ester VII is
synthesized from III in the same fashion as described above.
##STR00009## ##STR00010##
[0235] Another alternative route is illustrated in Scheme 2. As
described in Scheme 1a, boronic ester or acid, X, can be prepared
from aminopyridine or aminopyrimidine IX. Suzuki cross-coupling
reaction between compound X and pyridine or pyrazine XI then can
give the bi-heteroaryl intermediate XII. The SN.sub.AR reaction
between XII and functionalized amine HA.sub.4LR.sub.2 under basic
condition (DIEA, TEA, lutidine, pyridine) in a solvent such as DMF,
THF, DMSO, NMP, dioxane with heating (30-130.degree. C.) can give
compound V. When R.sub.1' is not identical to R.sub.1, further
functional manipulation will be needed to obtain VI. When R.sub.1'
is identical with R.sub.1, compound V will be the same as compound
VII.
##STR00011## ##STR00012##
[0236] Compounds of the present invention, listed in Table I, were
prepared by following the specific procedures outlined below. The
procedures include synthesis of intermediates and using these
intermediates to make compounds of Formula I.
[0237] Synthesis of Intermediates
Synthesis of 6-bromo-N-(3-fluorobenzyl) pyridin-2-amine
(Intermediate A)
##STR00013##
[0239] A solution of 2,6-dibromopyridine (7.1 g, 30.0 mmol) in NMP
(16 mL) was mixed with a mixture of (3-fluorophenyl)methanamine
(4.13 g, 33.0 mmol) and Huenig's Base (5.76 mL, 33.0 mmol). The
resulting mixture was stirred under argon at 115-120.degree. C. for
about 168 hr. The mixture was then cooled to ambient temperature
and diluted with EtOAc (250 mL). The organic layer was separated,
washed with saturated aqueous sodium bicarbonate (2.times.), water
(2.times.), brine (1.times.), dried over sodium sulfate, filtered,
and concentrated in vacuo to yield a crude material. The crude
material was purified by column chromatography [SiO.sub.2, 120 g,
EtOAc/hexane=0/100 to 20/80] providing 6-bromo-N-(3-fluorobenzyl)
pyridin-2-amine (7.11 g) as an off-white solid. LCMS (m/z):
281.1/283.1 [M+H]+; Retention time=1.03 min.
Synthesis of
5'-chloro-2'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine
##STR00014##
[0241] A mixture of 6-bromo-N-(3-fluorobenzyl)pyridin-2-amine (A,
2.0 g, 7.11 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid (2.0
g, 11.4 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (0.465 g,
0.569 mmol), DME (27 mL) and 2M aqueous Na.sub.2CO.sub.2 (9.25 mL,
18.50 mmol) was stirred at about 100.degree. C. for 3 hr. After
cooling to ambient temperature, the mixture was diluted with EtOAc
(25 mL) and MeOH (20 mL), filtered, and concentrated in vacuo to
yield a crude material. The crude material was purified by column
chromatography [silica gel, 120 g, EtOAc/hexane=0/100 to 20/80]
providing
5'-chloro-2'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine (1.26
g) as an off-white solid. LCMS (m/z): 332.2 [M+H]+; Retention
time=0.92 min.
Synthesis of
6-bromo-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(Intermediate C)
##STR00015##
[0243] A mixture of 2-bromo-6-fluoropyridine (750 mg, 4.26 mmol) in
DMSO (3 mL) was mixed with (tetrahydro-2H-pyran-4-yl)methanamine
hydrochloride (775 mg, 5.11 mmol) and NEt.sub.3 (1.426 mL, 10.23
mmol). The resulting mixture was heated at about 110.degree. C. for
18 hr. The mixture was cooled to ambient temperature and diluted
with EtOAc. The organic layer was separated, washed with saturated
aqueous sodium bicarbonate solution, water, and brine, dried over
sodium sulfate, filtered and concentrated in vacuo to yield a
resulting residue. The resulting residue was purified by column
chromatography [SiO.sub.2, 40 g, EtOAc/heptane=0/100 to 30/70].
Pure fractions were combined and concentrated in vacuo providing
6-bromo-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine (B1,
940 mg) as a white solid. LCMS (m/z): 271.0/272.9 [M+H]+; Retention
time=0.81 min.
Synthesis of
5'-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridin-6-
-amine (Intermediate D)
##STR00016##
[0245] A mixture of
6-bromo-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine (C, 271
mg, 1 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid (351 mg,
2.000 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (82 mg, 0.100
mmol) in DME (4.5 mL) and 2M Na.sub.2CO.sub.3 (318 mg, 3.00 mmol)
was heated in a sealed tube at about 103.degree. C. for about 2 hr.
The mixture then was cooled to ambient temperature, diluted with
EtOAc (.about.25 mL) and MeOH (.about.5 mL), filtered, and
concentrated in vacuo to yield a resulting residue. The resulting
residue was purified by column chromatography [SiO.sub.2, 12 g,
EtOAc/heptane=10/90 to 50/50]. Fractions were combined and
concentrated in vacuo providing
5'-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridin-6-
-amine (260 mg) as a yellow thick oil. LCMS (m/z): 322.1/323.9
[M+H]+; Retention time=0.60 min.
Synthesis of
6-bromo-5-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(E) and
6-bromo-3-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(Intermediate F)
##STR00017##
[0247] A solution of
6-bromo-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine (C,
1000 mg, 3.69 mmol) in chloroform (15 mL) was diluted with
1-chloropyrrolidine-2,5-dione (NCS, 492 mg, 3.69 mmol). The mixture
then was heated in a sealed tube at about 33.degree. C. for about
16 hr, followed by heating the reaction mixture for about 24 hr at
about 37.degree. C., and then for an additional 5 days at about
43.degree. C. The reaction mixture then was cooled to ambient
temperature, diluted with 1N aqueous sodium hydroxide solution and
DCM. The organic layer was separated, washed with brine, dried over
sodium sulfate, filtered off and concentrated in vacuo. The
resulting residue was purified by column chromatography [ISCO.
SiO2, 80 g, EtOAc/heptane=5/95 2 min, 5/95 to 30/70 2-15 min, to
35/65 15-18 min, then 35%]. Fractions were combined and
concentrated in vacuo yielding
6-bromo-3-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(F, 453 mg), and
6-bromo-5-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(E, .about.500 mg). (F): LCMS (m/z): 305.0 [M+H]+; Retention
time=1.01 min. (E): LCMS (m/z): 305.0 [M+H]+; Retention time=0.96
min.
Synthesis of
3,5'-dichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine (Intermediate G)
##STR00018##
[0249] A mixture of
6-bromo-5-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(E, 300 mg, 0.982 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid
(344 mg, 1.963 mmol), PdCl2(dppf).CH2Cl2 adduct (80 mg, 0.098 mmol)
in DME (4.5 mL) and 2M aqueous sodium carbonate (4.5 mL, 4.50 mmol)
was heated in a sealed tube at about 103.degree. C. for about 16
hr. The reaction mixture was cooled to ambient temperature, diluted
with EtOAc (.about.100 mL) and saturated aqueous sodium carbonate
solution. The organic layer was separated, washed with saturated
aqueous sodium carbonate solution (2.times.), dried over sodium
sulfate, filtered off and concentrated in vacuo. The resulting
residue was purified by column chromatography [ISCO, SiO2, 25 g,
EtOAc/heptane=0/100 to 25/75]. Fractions were combined and
concentrated in vacuo providing
3,5'-dichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine (140 mg) as a light brown liquid. LCMS (m/z): 356.1
[M+H]+; Retention time=0.96 min.
Synthesis of
5,5'-dichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine (Intermediate H)
##STR00019##
[0251] A mixture of
6-bromo-3-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(F, 200 mg, 0.654 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid
(230 mg, 1.309 mmol), PdCl2(dppf).CH2Cl2 adduct (53.4 mg, 0.065
mmol) in DME (3 mL) and 2M aqueous sodium carbonate (3 mL, 6.00
mmol) was heated in a sealed tube at about 103.degree. C. for 16
hr. The reaction mixture was cooled to ambient temperature, diluted
with EtOAc (.about.100 mL) and saturated aqueous sodium bicarbonate
solution. The organic layer was separated, washed with saturated
aqueous sodium bicarbonate solution (2.times.), dried over sodium
sulfate, filtered off and concentrated in vacuo. The resulting
residue was purified by column chromatography [ISCO, SiO2, 25 g,
EtOAc/heptane=0/100 to 30/70]. Fractions were combined and
concentrated in vacuo providing
5,5'-dichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine (130 mg) as a nearly colorless liquid. LCMS (m/z): 356.1
[M+H]+; Retention time=1.10 min.
Synthesis of
5'-chloro-2',5-difluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine (Intermediate I)
##STR00020##
[0252] Step 1. Preparation of
3,6-difluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
[0253] A mixture of 2,3,6-trifluoropyridine (3 g, 22.54 mmol),
(tetrahydro-2H-pyran-4-yl)methanamine (3.89 g, 33.8 mmol) and
triethylamine (7.86 mL, 56.4 mmol) in NMP (60 mL) was heated at
about 70.degree. C. for about 1 hr. The reaction mixture was cooled
to ambient temperature, diluted with EtOAc (.about.100 mL), brine
(.about.50 mL) and water (.about.50 mL). The separated organic
layer was washed with brine (1.times.), 0.3N aqueous HCl
(2.times.), saturated aqueous NaHCO.sub.3 solution (1.times.),
brine (1.times.), dried over Na.sub.2SO.sub.4, filtered off and
concentrated in vacuo providing crude
3,6-difluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine,
which was directly used in the next reaction without further
purification. Yield: 3.5 g. LCMS (m/z): 229.1 [M+H]+; Retention
time=0.79 min.
Step 2. Preparation of
3-fluoro-6-methoxy-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
[0254] To a solution of
3,6-difluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine (5
g, 21.91 mmol) in MeOH (35 mL) was added sodium methoxide (25 wt. %
in MeOH, 15.03 mL, 65.7 mmol). The resulting mixture was heated in
a steel bomb at about 135.degree. C. for .about.18 hr. The mixture
then was cooled to ambient temperature and concentrated in vacuo.
The resulting residue was taken up in water (.about.250 mL)
yielding a precipitate, which was collected by filtration, and then
washed with water. The solid then was dissolved in toluene (10
mL)/DCM (10 mL), decanted from the dark brownish film and
concentrated in vacuo. The resulting residue was dried in high
vacuo providing crude
3-fluoro-6-methoxy-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
as a nearly colorless oil, which was directly used in the next
reaction without further purification. Yield: 4.96 g. LCMS (m/z):
241.1 [M+H]+; Retention time=0.87 min.
Step 3. Preparation of
5-fluoro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyridin-2-ol
[0255] To a solution of
3-fluoro-6-methoxy-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
[0256] (4.6 g, 19.14 mmol) in acetonitrile (50 mL) was added sodium
iodide (20.09 g, 134 mmol) and TMS-chloride (17.13 mL, 134 mmol).
The resulting mixture was stirred at about 95.degree. C. for 20 hr.
The reaction mixture was cooled to ambient temperature and then
diluted with EtOAc (80 mL) and water (40 mL). The diluted mixture
was stirred vigorously for about 30 min. The organic layer was
separated and washed with 0.1N aqueous HCl solution. The combined
aqueous layers were carefully neutralized (pH .about.7) with solid
NaHCO.sub.3 solution and extracted with EtOAc (1.times.100 mL) and
DCM (2.times.50 mL). The combined organic layers were washed with
saturated aqueous NaHCO.sub.3 solution and brine, dried over
Na.sub.2SO.sub.4, filtered off and concentrated in vacuo. The
resulting residue was purified by column chromatography [SiO.sub.2,
80 g, EtOAc/heptane=10/90 for 2 min, EtOAc/heptane=10/90 to 100/0
over 23 min, then EtOAc/heptane=100/0] providing
5-fluoro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyridin-2-ol as a
highly viscous oil which turned to purple upon standing at room
temperature. Yield: 780 mg. LCMS (m/z): 227.1 [M+H]+; Retention
time=0.42 min.
Step 4. Preparation of
5-fluoro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyridin-2-yl
trifluoromethanesulfonate
[0257] A solution of
5-fluoro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyridin-2-ol (500
mg, 2.210 mmol) and triethylamine (0.462 mL, 3.31 mmol) in DCM (20
m L) was gradually diluted at about 0.degree. C. with
trifluoromethanesulfonic anhydride (1.120 mL, 6.63 mmol). The
resulting mixture was stirred for about 2 hr at 0.degree. C. and
carefully mixed with ice-cooled saturated aqueous NaHCO; solution.
The aqueous layer was separated, and extracted with DCM (2.times.).
The combined organic layers were dried over Na.sub.2SO.sub.4,
filtered off and concentrated in vacuo. The resulting residue was
purified by column chromatography [SiO.sub.2, 40 g, 30 min,
EtOAc/heptane=5/95 to 40/60] providing
5-fluoro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyridin-2-yl
trifluoromethanesulfonate as a colorless oil. Yield: 743 mg. LCMS
(m/z): 359.0 [M+H]+; Retention time=1.02 min.
Step 5. Preparation of
5'-chloro-2',5-difluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine
[0258] A mixture of
5-fluoro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyridin-2-yl
trifluoromethanesulfonate (712 mg, 1.987 mmol),
5-chloro-2-fluoropyridin-4-ylboronic acid (697 mg, 3.97 mmol),
PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (162 mg, 0.199 mmol) in
DME (8 mL) and 2 M aqueous Na.sub.2CO.sub.3 solution (2.6 mL, 1.987
mmol) in a sealed tube was heated at 95.degree. C. for 3 hr. The
mixture was allowed to cool to ambient temperature and was diluted
with EtOAc (.about.100 mL) and saturated aqueous NaHCO.sub.3
solution. The separated organic layer was washed with saturated
aqueous NaHCO.sub.3 (2.times.), dried over Na.sub.2SO.sub.4,
filtered off and concentrated in vacuo. The resulting resulting
residue was purified by column chromatography [SiO.sub.2, 40 g,
EtOAc/heptane=0/100 to 25/75 over 20 min] providing
5'-chloro-2',5-difluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine as a white solid. Yield: 570 mg. LCMS (m/z): 340.1
[M+H]+; Retention time=0.99 min.
Synthesis of
(R/S)-5'-chloro-N-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)-2'-fluor-
o-2,4'-bipyridin-6-amine (Intermediate J)
##STR00021##
[0259] Step 1. Preparation of tert-butyl
6-bromopyridin-2-ylcarbamate
[0260] To a solution of 6-bromopyridin-2-amine (3 g, 17.34 mmol),
triethylamine (3.14 mL, 22.54 mmol) and DMAP (0.424 g, 3.47 mmol)
in DCM (24 mL) was added slowly a solution of BOC-anhydride (4.83
mL, 20.81 mmol) in DCM (6 mL). The reaction mixture was stirred at
ambient temperature for .about.24 hr. The mixture was diluted with
water, brine and EtOAc. The separated aqueous layer was extracted
with EtOAc. The combined organic layers were dried over sodium
sulfate and concentrated in vacuo. The resulting residue was
purified by column chromatography providing tert-butyl
6-bromopyridin-2-ylcarbamate as a white solid. Yield: 1.67 g. LCMS
(m/z): 274.9 [M+H]+; Retention time=0.95 min.
Step 2: Preparation of
(R/S)-(2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl
4-methylbenzenesulfonate
[0261] To a solution of
(2,2-dimethyltetrahydro-2H-pyran-4-yl)methanol (1 g, 6.93 mmol) in
DCM (5 mL) and pyridine (5 mL, 61.8 mmol) was added
para-toluenesulfonyl chloride (1.586 g, 8.32 mmol) and DMAP (0.042
g, 0.347 mmol). The mixture was stirred for 18 hr at ambient
temperature. The reaction mixture was concentrated in vacuo and the
resulting resulting residue was diluted with water and DCM. The
separated organic layer was washed with 0.2N aqueous HCl
(1.times.), 1N aqueous HCl (2.times.), brine, dried over sodium
sulfate, filtered off and concentrated in vacuo. The resulting
residue was purified by column chromatography [SiO.sub.2, 40 g,
EtOAc/hexane=0/100 to 50/50; 25 min] providing
(R/S)-(2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl
4-methylbenzenesulfonate as a colorless oil. Yield: 2.05 g. LCMS
(m/z): 299.1 [M+H]+; Retention time=0.96 min.
Step 3: Preparation of (R/S)-tert-butyl
6-bromopyridin-2-yl((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)carbamat-
e
[0262] To a mixture of tert-butyl 6-bromopyridin-2-ylcarbamate (686
mg, 2.51 mmol), K.sub.2CO.sub.3 (347 mg, 2.51 mmol),
(2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl
4-methylbenzenesulfonate (750 mg, 2.51 mmol) in DMF (10 mL) was
added carefully NaH (60 wt. %, 141 mg, 3.52 mmol) in portions
[Caution: gas development!]. The resulting mixture was stirred at
about 45.degree. C. for 4 hr. The mixture was warmed to ambient
temperature and was diluted with EtOAc (.about.50 mL) and saturated
aqueous NaHCO.sub.3. The organic layer was separated, washed with
saturated aqueous NaHCO.sub.3 solution (1.times.), dried over
Na.sub.2SO.sub.4, filtered off and concentrated in vacuo. The
resulting residue was purified by column chromatography [SiO.sub.2,
40 g, 25 min, EtOAc/heptane=0/100 to 25/75 over 25 min] providing
(R/S)-tert-butyl
6-bromopyridin-2-yl((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)carbamat-
e as highly viscous, colorless oil. Yield: 723 mg. LCMS (m/z):
344.9 {loss of tert Bu-group}/(399.0). [M+H]+; Retention time=1.22
min.
Step 4: Preparation of (R/S)-tert-butyl
5'-chloro-2'-fluoro-2,4'-bipyridin-6-yl((2,2-dimethyltetrahydro-2H-pyran--
4-yl)methyl)carbamate
[0263] A mixture of tert-butyl
6-bromopyridin-2-yl((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)carbamat-
e (710 mg, 1.778 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid,
PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (145 mg, 0.178 mmol) in
DME (7 mL) and 2M aqueous Na.sub.2CO.sub.3 solution (2.3 mL, 1.778
mmol) was heated in a sealed tube at about 98.degree. C. for 2 hr.
The mixture was cooled to ambient temperature and diluted with
EtOAc (.about.100 mL) and saturated aqueous NaHCO.sub.3 solution.
The separated organic layer was washed with saturated aqueous
NaHCO.sub.3 (2.times.), dried over Na.sub.2SO.sub.4, filtered off
and concentrated in vacuo. The resulting residue was purified by
column chromatography [SiO.sub.2, 40 g, 25 min, EtOAc/heptane=0/100
to 25/75 over 25 min] providing (R/S)-tert-butyl
5'-chloro-2'-fluoro-2,4'-bipyridin-6-yl((2,2-dimethyltetrahydro-2H-pyran--
4-yl)methyl)carbamate as a highly viscous, colorless oil. Yield:
605 mg. LCMS (m/z): 394.1 {loss of tert Bu-group}/450.2 [M+H]+;
Retention time=1.24 min.
Step 5. Preparation of
(R/S)-5'-chloro-N-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)-2'-fluor-
o-2,4'-bipyridin-6-amine
[0264] To a solution of tert-butyl
5'-chloro-2'-fluoro-2,4'-bipyridin-6-yl((2,2-dimethyltetrahydro-2H-pyran--
4-yl)methyl)carbamate (950 mg, 2.111 mmol) in methanol (5 mL) was
added 4M HCl/dioxane (15 mL, 494 mmol). The resulting mixture was
stirred for .about.45 min at ambient temperature. The mixture then
was concentrated in vacuo and the resulting residue was dissolved
in EtOAc (.about.50 mL) and saturated aqueous NaHCO.sub.3 solution
(.about.50 mL). The separated organic layer was washed with
saturated aqueous NaHCO.sub.3 solution (1.times.), brine
(1.times.), dried over Na.sub.2SO.sub.4, filtered off and
concentrated in vacuo providing crude
(R/S)-5'-chloro-N-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)-2'-fluor-
o-2,4'-bipyridin-6-amine as a colorless oil, which was directly
used in the next reaction without further purification. Yield: 740
mg. LCMS (m/z): 350.1 [M+H]+; Retention time=0.69 min.
Synthesis of 5'-chloro-2',3,6-trifluoro-2,4'-bipyridine
(Intermediate K)
##STR00022##
[0265] Step 1. Preparation of 3,6-difluoro-2-methoxypyridine
[0266] 2,3,6-Trifluoropyridine (17.91 ml, 188 mmol) was dissolved
in anhydrous MeOH (300 ml) and the resulting mixture was placed
under argon. This mixture then was treated with a 25 wt %
methanolic solution of sodium methoxide (43.0 ml, 188 mmol). The
resulting mixture was then heated at about 65.degree. C. for 2 hr.
The reaction mixture was cooled to ambient temperature, and
concentrated in vacuo to yield a residue which then was mixed with
brine (200 mL), and extracted with Et2O (3.times.200 ml). The
combined extracts were dried (Na2SO4), filtered, and concentrated
in vacuo to give 21.5 g (79% yield) of crude
3,6-difluoro-2-methoxypyridine as a white solid which was carried
on to the next step without purification.
Step 2. Preparation of 3,6-difluoro-2-hydroxypyridine
[0267] To 3,6-difluoro-2-methoxypyridine (21.5 g, 148 mmol) in
acetonitrile (250 ml) was added sodium iodide (66.6 g, 445 mmol)
and chlorotrimethylsilane (56.8 ml, 445 mmol). The resulting
mixture was heated at 80-85.degree. C. for 2.5 hr. The mixture was
cooled to ambient temperature and diluted with EtOAc (300 mL) and
water (300 mL) and vigorously stirred for another hr. The layers
were separated, and the aqueous phase was extracted with additional
ethyl acetate (200 mL). The combined organic layers were washed
sequentially with 0.6 N aqueous HCl (250 mL) and brine (250 mL) and
concentrated in vacuo to yield a slurry. The slurry was filtered
and rinsed three times with cold acetonitrile to yield 10.8 g of
desired product as a white solid. The filtrate was concentrated and
purified by flash chromatography over silica gel (heptanes:ethyl
acetate gradient) to give an additional 4.2 g (77% yield combined)
of 3,6-difluoro-2-hydroxypyridine as a white solid. LCMS (m/z):
132.0 [M+H]+; retention time=0.47 min.
Step 3. Preparation of 3,6-difluoropyridin-2-yl
trifluoromethanesulfonate
[0268] An ice water bath-cooled solution of
3,6-difluoro-2-hydroxypyridine (10.75 g, 82 mmol) and triethylamine
(22.86 ml, 164 mmol) in DCM (550 ml) was mixed with a solution of
trifluoromethanesulfonic anhydride (16.63 ml, 98 mmol) in DCM (100
ml) over 20 min. The resulting mixture then was stirred for 2 hr at
0.degree. C., with the progress of the reaction followed by TLC
(2:1 heptanes:ethyl acetate). The reaction mixture was quenched
with saturated aqueous NaHCO3 solution (200 mL). The separated
aqueous layer was extracted with DCM (2.times.). The combined
organic layers were dried over sodium sulfate, filtered, and
concentrated in vacuo. The resulting residue was purified by column
chromatography over silica gel (EtOAc/heptane gradient) to give
16.3 g (76% yield) of 3,6-difluoropyridin-2-yl
trifluoromethanesulfonate as a yellow oil.
Step 4. Preparation of
5'-chloro-2',3,6-trifluoro-2,4'-bipyridine
[0269] A mixture of 3,6-difluoropyridin-2-yl
trifluoromethanesulfonate (3.50 g, 13.30 mmol) and
5-chloro-2-fluoropyridine-4-boronic acid (3.27 g, 18.62 mmol) in
THF (27 ml) was degassed by bubbling Argon gas for 10 min. Aqueous
sodium carbonate (13.30 ml, 26.6 mmol) and
PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (0.652 g, 0.798 mmol) were
added, and the mixture was degassed for an additional 5 min. The
resulting reaction mixture was stirred at about 100.degree. C. for
2 hr in a sealed vessel. The reaction mixture was cooled to ambient
temperature, diluted with EtOAc and water. The separated organic
layer was dried over Na.sub.2SO.sub.4, filtered, and concentrated
in vacuo. The resulting residue was purified by column
chromatography over silica gel (heptanes/ethyl acetate gradient) to
yield 2.78 g (85% yield) of
5'-chloro-2',3,6-trifluoro-2,4'-bipyridine as a crystalline solid.
LCMS (m/z): 244.9 [M+H]+; retention time=0.86 min.
Synthesis of
5'-chloro-N-(((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)methyl)-2'-flu-
oro-2,4'-bipyridin-6-amine (Intermediate L)
##STR00023##
[0270] Step 1. Preparation of
(2R,6S)-2,6-dimethyldihydro-2H-pyran-4(3H)-one
[0271] A solution of 2,6-dimethyl-4H-pyran-4-one (2 g, 16.1 mmol)
in 20 ml ethanol was stirred over 10% Pd/C (0.2 g) under hydrogen
(15 psi) for 16 hours at ambient temperature. TLC showed two spots;
one was desired product and second one was side product in a 1:1
ratio. GCMS M+ 128 for product, and M+ 130 for side product.
[0272] Suspension was filtered off, and the filtrate was
concentrated to remove solvent to give 2.3 g crude product which
contained .about.30% of the side product. The resulting oily
residue was treated with 2.3 g Dess-Martin periodinane in 15 ml DCM
at ambient temperature for 16 hours. GCMS showed oxidation was
complete, desired product formation was confirmed by GCMS at M+
128. .about.3 ml NaS2CO3 was added to the suspension and the
resulting mixture was stirred for 1 hour at ambient temperature,
then 20 ml saturated sodium bicarbonate solution was added to, and
new mixture was stirred for another hour. The organic phase was
separated, washed with water, brine, dried and filtered through
celite. The filtrate was concentrated and resulting residue was
purified by ISCO eluting with 10% ethyl acetate in heptane to yield
600 mg of the desired product. GCMS: M=128. HNMR: 1.5 ppm (6H), 2.3
ppm (4H), 3.75 ppm (2H).
Step 2. Preparation of
(2R,6S,E)-4-(methoxymethylene)-2,6-dimethyltetrahydro-2H-pyran
[0273] To a suspension of (methoxymethyl)triphenyl phosphine
chloride (1.5 g, 4.45 mmol) in 8 ml THF at -10.degree. C., was
added dropwise 4.45 ml 1.0M/THF solution of sodium
bis(trimethylsilyl)amide. The resulting reaction mixture was
stirred for 1 hour, followed by addition of a solution of
(2R,6S)-2,6-dimethyldihydro-2H-pyran-4(3H)-one (380 mg, 2.96 mmol)
in 2 ml THF. The resulting mixture was warmed to ambient
temperature and stirred for an additional 3 hours. GCMS showed
formation of desired product at M+156, as major component. The
reaction mixture was quenched with 15 ml water, and was extracted
with diethyl ether (2.times.30 ml). The combined organic phase was
washed with brine, dried and concentrated. The resulting residue
was purified by ISCO eluting with 10% ethyl acetate in heptane to
yield 240 mg of the desired product as a colorless oil, GCMS showed
M=156. HNMR: 5.9 ppm (1H), 3.45 ppm (3H), 3.25 ppm (2H), 2.45 ppm
(1H), 1.85 ppm (1H), 1.6 ppm (1H), 1.38 ppm (1H), 1.1 ppm (6H).
Step 3. Preparation of
(2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-carbaldehyde
[0274] A mixture of
(2R,6S,E)-4-(methoxymethylene)-2,6-dimethyltetrahydro-2H-pyran (240
mg, 1.53 mmol) and 88% formic acid (1.5 ml, 34.4 mmol) in water was
heated in an oil bath under Argon to about 90.degree. C. for 1
hour. GCMS indicated that reaction was complete under the
condition. The reaction mixture was cooled in an ice bath,
neutralised with 6N NaOH to a pH=6, and extracted with diethyl
ether. The organic phase were dried and concentrated to dryness to
yield 120 mg of the desired product as yellow colored oil. GCMS
M=142. HNMR showed 9.51 ppm (s, 1H, CHO).
Step 4. Preparation of
6-bromo-N4((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)methyl)
pyridin-2-amine
[0275] The mixture of
(2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-carbaldehyde (120 mg,
0.84 mmol) and 6-bromo-2-aminopyridine (219 mg, 1.26 mmol) in 5 ml
DCM was stirred at ambient temperature for about 40 min. To this
solution was added sodium triacetoxy borohydride (268 mg, 1.26
mmol), followed by the addition of 0.01 ml acetic acid. The
resulting solution was stirred at ambient temperature for about 40
hours. The reaction mixture was concentrated in vacuo to yield a
residue was diluted with ethyl acetate, washed with sodium
bicarbonate, brine, dried, concentrated. The resulting residue was
purified by ISCO eluting with 10% to 20% ethyl acetate in heptane
to yield 110 mg of the desired product as colorless oil. LCMS
(m/z): 299/301 (MH+), retention time=1.01 min.
Step 5. Preparation of
5'-chloro-N-(((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)methyl)-2'-flu-
oro-2,4'-bipyridin-6-amine
[0276] A mixture of
6-bromo-N-(((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)methyl)
pyridin-2-amine (110 mg, 0.36 mmol),
5-chloro-2-fluoro-pyridine-4-boronic acid (193 mg, 1.10 mmol), 0.55
ml 2.0M saturated sodium carbonate aqueous solution in 2 ml DME was
purged with Argon for 3 min, PdCl.sub.2(dppf)CH.sub.2Cl.sub.2 (30
mg, 0.037 mmol) was added to this purged. The resulting mixture was
heated at about 95.degree. C. in an oil bath for 3.5 hours.
Formation of the desired product was confirmed by LCMS: MH+ 350,
0.70 min. The preceding reaction mixture was diluted with ethyl
acetate, washed with water, brine, dried over sodium sulfate and
concentrated. The resulting residue was purified by ISCO eluting
with 10% ethyl acetate in heptane to give 90 mg desired product as
colorless oil. LCMS (m/z): 350 (MH+), retention time=0.70 min.
Synthesis of
5'-chloro-N6-(((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)methyl)-2,4'--
bipyridine-2',6-diamine (Intermediate M)
##STR00024##
[0278] A mixture of
5'-chloro-N-(((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)methyl)-2'-flu-
oro-2,4'-bipyridin-6-amine (60 mg, 0.17 mmol), and 3.0 ml 28%
ammonium hydroxide aqueous solution was heated at about 130.degree.
C. in an oil bath for 17 hours. Formation of compound M was
Reaction confirmed by LCMS/LC data. The reaction mixture was
diluted with ethyl acetate, washed with water, saturated sodium
bicarbonate, and brine, dried over sodium sulfate and concentrated
to yield 50 mg of the desired product. LCMS (m/z): 347 (MH+),
retention time=0.53 min.
Synthesis of
3-bromo-5'-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bip-
yridin-6-amine (Intermediate N)
##STR00025##
[0280] A mixture of
5'-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridin-6-
-amine (516 mg, 1.60 mmol) and N-bromosuccinimide (286 mg, 1.60
mmol) in acetonitrile (12 mL) was stirred at 90.degree. C. for 3 hr
in a sealed vessel. Volatiles were removed under reduced pressure.
The resulting residue was dissolved in ethyl acetate and washed
sequentially with saturated aqueous sodium bicarbonate and brine.
The organic phase was dried (Na2SO4), filtered, and concentrated.
The crude material was purified by column chromatography over
silica gel (heptanes/ethyl acetate gradient) to yield 608 mg of the
desired product. LCMS (m/z): 402.0 [M+H]+; Retention time=1.03
min.
Synthesis of intermediate (4-methoxytetrahydro-2H-pyran-4-yl)
methyl 4-methylbenzenesulfonate (Intermediate O)
##STR00026##
[0281] Step 1. Synthesis of 1,6-dioxaspiro[2.5]octane
[0282] To a clear solution of trimethylsulfonium iodide (3.27 g, 16
mmol) in 20 ml of DMSO was added dihydro-2H-pyran-4(3H)-one (1.0 g,
10 mmol) with stirring. To this mixture, under nitrogen, was then
slowly added KO.sup.tBu (1.68 g, 15 mmol) in 15 ml of DMSO. The
resulting solution was then stirred overnight at ambient
temperature. Water (50 ml) was slowly added to the mixture, and the
resulting mixture was extracted with diethyl ether (3.times.20 ml).
The ether layers were combined, dried and concentrated in vacuo to
yield 650 mg of the crude product. 1H NMR (300 MHz, CHLOROFORM-d)
.delta. ppm 1.44-1.62 (m, 2H) 1.76-1.98 (m, 2H) 2.70 (s, 2H)
3.70-3.98 (m, 4H).
Step 2. Synthesis of (4-methoxytetrahydro-2H-pyran-4-yl)
methanol
[0283] To a solution of 1,6-dioxaspiro[2.5]octane (600 mg, 5.26
mmol) in methanol (10 ml) at 0.degree. C. (ice-water) under
nitrogen was added camphorsulfonic acid (50 mg, 0.21 mmol) and the
resulting mixture was stirred at about 0.degree. C. for 2 hours.
The mixture was concentrated in vacuo and the crude residue was
used in the next step without purification. The desired product was
obtained as a light yellow oil (707 mg).
Step 3
[0284] To a solution of (4-methoxytetrahydro-2H-pyran-4-yl)
methanol (300 mg, 2.05 mmol) in pyridine (4 ml) at ambient
temperature was added toluenesulfonic chloride (430 mg, 2.25 mmol)
and the resulting mixture was stirred overnight at about 25.degree.
C. The stirred mixture was concentrated and the solid residue was
dissolved in DCM and purified by silica gel chromatography using a
12 g column, eluting with 0-30% ethyl acetate in heptane to yield
the desired compound "O" as a light yellow solid (360 mg). 1H NMR
(300 MHz, CHLOROFORM-d) .delta. ppm 1.45-1.63 (m, 2H) 1.61-1.79 (m,
2H) 2.46 (s, 3H), 3.16 (s, 3H) 3.53-3.75 (m, 4H) 3.93 (s, 2H), 7.36
(d, J=8.20 Hz, 2H) 7.81 (d, J=8.20 Hz, 2H).
Synthesis of tert-butyl
6-bromo-5-chloropyridin-2-yl((4-methoxytetrahydro-2H-pyran-4-yl)methyl)ca-
rbamate (Intermediate P)
##STR00027##
[0286] To a stirred solution of tert-butyl
6-bromo-5-chloropyridin-2-ylcarbamate (140 mg, 0.455 mmol) in DMF
(2 ml) under nitrogen was added NaH (60%, 30 mg, 0.774 mmol). The
resulting mixture was stirred at ambient temperature for one hour.
A solution of (4-methoxytetrahydro-2H-pyran-4-yl)methyl
4-methylbenzenesulfonate (intermediate O, 164 mg, 0.546 mmol) in
DMF (1.5 ml) was then added to the preceding mixture. The resulting
mixture was then stirred overnight at about 85.degree. C. The
stirred mixture was diluted with 30 ml of ethyl acetate, washed
with water (20 ml.times.3) and dried. After concentration the
resulting residue was purified by silica gel chromatography using a
12 g column, eluting with 5-20% ethyl acetate in hexane to yield
the desired compound "P" as a viscous oil (92 mg), which solidified
upon standing overnight. LCMS (m/z): 437.0 [M+H]+; Retention
time=1.158 min.
Synthesis of (1-methoxycyclohexyl)methyl 4-methylbenzenesulfonate
(Intermediate Q)
##STR00028##
[0288] This compound was synthesized from cyclohexanone following
the procedure described for
(4-methoxytetrahydro-2H-pyran-4-yl)methyl 4-methylbenzenesulfonate
(Intermediate O).
[0289] LCMS (m/z): 299.2 [M+H]+; Retention time=1.055 min.
Synthesis of 4-(aminomethyl)tetrahydro-2H-pyran-4-carbonitrile
(Intermediate R)
##STR00029##
[0290] Step 1. Synthesis of
dihydro-2H-pyran-4,4(3H)-dicarbonitrile
[0291] A mixture of malononitrile (0.991 g, 15 mmol),
1-bromo-2-(2-bromoethoxy)ethane (3.83 g, 16.50 mmol) and DBU (4.97
ml, 33.0 mmol) in DMF (6 ml) was heated at about 85.degree. C. for
3 hours, and then cooled to ambient temperature. The mixture was
concentrated in vacuo, the resulting residue was diluted with ethyl
acetate, washed three times with water and dried overnight under
high vacuum to yield the desired product as a light brown solid
(1.65 g). GC-MS: 136 [M]; Retention time=5.76 min. 1H NMR (300 MHz,
CHLOROFORM-d) .delta. ppm 2.14-2.32 (m, 4H) 3.77-3.96 (m, 4H).
Step 2
[0292] A mixture of dihydro-2H-pyran-4,4(3H)-dicarbonitrile (450
mg, 3.31 mmol)<autotext key="0BD391A6" name="[Reactants]"
index="1" field="Reactants" type="field" length="34"/> and
Sodium borohydride (375 mg, 9.92 mmol)<autotext key="0BD391A7"
name="[Reactants]" index="2" field="Reactants" type="field"
length="38"/> in EtOH (15 ml)<autotext key="0BD391 A8"
name="[Solvents]" index="1" field="Solvents" type="field"
length="12"/> was stirred at ambient temperature for about 4
hours. The mixture was concentrated and the resulting residue was
diluted with ethyl acetate, washed with water and dried.
Concentration in vacuo afforded 388 mg of the crude product which
was used directly in the next step. LCMS (m/z): 141.0 [M+H]+;
Retention time=0.18 min.
Synthesis of
4-((6-bromopyridin-2-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile
(Intermediate S)
##STR00030##
[0294] To 2-bromo-6-fluoropyridine (400 mg, 2.273 mmol) in DMSO (4
ml) at ambient temperature was sequentially added
4-(aminomethyl)tetrahydro-2H-pyran-4-carbonitrile (Intermediate R.
382 mg, 2.73 mmol) and triethylamine (0.792 ml, 5.68 mmol). The
resulting light brown mixture was heated at 110.degree. C. in a
sealed glass bomb for 18 hours. The reaction mixture then was
cooled to ambient temperature, reaction mixture diluted with EtOAc,
washed with saturated NaHCO3 solution and brine, dried over sodium
sulfate and concentrated in vacuo to yield 890 mg of a light brown
liquid. The crude material was purified by silica gel
chromatography using a 12 g column, eluting with 5%-20% ethyl
acetate in hexane to afford 410 mg (60.9%) of the desired product
"S". LCMS (m/z): 297.9 [M+H]+; Retention time=0.823 min. 1H NMR
(400 MHz, CHLOROFORM-d) .delta. ppm: 1.67-1.96 (m, 4H), 3.59-3.78
(m, 4H), 3.98 (m, 2H), 4.82 (t, J=6.65 Hz, 1H), 6.39 (d, J=8.22,
1H), 6.72-6.84 (m, 1H), 7.16-7.33 (m, 1H).
Synthesis of
5'-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-5-(trifluorometh-
yl)-2,4'-bipyridin-6-amine (Intermediate T) and
5'-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-3-(trifluorometh-
yl)-2,4'-bipyridin-6-amine (Intermediate U)
##STR00031##
[0295] Step 1. Synthesis of
6-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)-5-(trifluoromethyl)pyridin--
2-amine and
6-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)-3-(trifluoromethyl)pyridin--
2-amine
##STR00032##
[0297] To a solution of 2,6-dichloro-3-(trifluoromethyl)pyridine
(320 mg, 1.482 mmol) in DMSO (1.5 ml) at ambient temperature was
added (tetrahydro-2H-pyran-4-yl)methanamine (188 mg, 1.630 mmol)
and triethylamine (0.207 ml, 1.482 mmol). The resulting light brown
mixture was heated at about 120.degree. C. in a sealed glass bomb
for about 18 hours. The reaction mixture was cooled to ambient
temperature, diluted with EtOAc (20 mL), washed with saturated
NaHCO.sub.3 solution and brine, dried over sodium sulfate and
concentrated in vacuo to yield 502 mg of a light brown crude
liquid, which was purified by column chromatography (5 to 50% ethyl
acetate in heptane) to yield the desired products.
[0298]
6-chloro-N-(tetrahydro-2H-pyran-4-yl)methyl)-5-(trifluoromethyl)pyr-
idin-2-amine: 340 mg, 78%: LCMS (m/z): 295.2 [M+H]+; Retention
time=0.971 min; and
6-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)-3-(trifluoromethyl-
)pyridin-2-amine: 80 mg, 18%. LCMS (m/z): 295.1 [M+H]+; Retention
time=1.033 min.
Step 2a
[0299] A mixture of
6-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)-3-(trifluoromethyl)pyridin--
2-amine (100 mg, 0.339 mmol), 5-chloro-2-fluoropyridin-4-ylboronic
acid (89 mg, 0.509 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct
(27.7 mg, 0.034 mmol), DME (1.5 mL) and 2M aqueous Na.sub.2CO.sub.2
(0.5 mL, 1 mmol) was stirred in a sealed glass vessel at about
100.degree. C. for about 3 hours. After cooling to ambient
temperature the mixture was diluted with EtOAc (25 mL) and MeOH (20
mL), filtered and concentrated in vacuo. The resulting crude
material was purified by column chromatography [silica gel, 12 g,
EtOAc/hexane=5/100 to 50/50] to yield
5'-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-5-(trifluorometh-
yl)-2,4'-bipyridin-6-amine (Intermediate T, 102 mg, 77%). LCMS
(m/z): 390.2 [M+H]+; Retention time=1.12 min.
Step 2b
[0300] Intermediate U was synthesized following the procedure
described for
5'-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-5-(trifluoro-
methyl)-2,4'-bipyridin-6-amine
[0301] LCMS (m/z): 390.2 [M+H]+; Retention time=1.01 min.
Synthesis of
3,5'-dichloro-N-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)-2'-fluoro--
2,4'-bipyridin-6-amine (Intermediate V)
##STR00033##
[0303] Step 1. 6-Bromo-2-aminopyridine (15 g, 87 mmol) and TEA
(13.3 mL, 95 mmol) were dissolved in 173 mL of DCM. BOC-anhydride
(20.8 g, 95 mmol) was then dissolved in 100 mL of DCM and added
over 10 min using a syringe pump. The reaction mixture was stirred
at ambient temperature for 72 hr. The solvents were evaporated and
the resulting residue was purified by silica gel chromatography
(heptane:EtOAc 1:0 to 7:3) to give the product as a colorless solid
(23.0 g, 97%). LCMS (m/z): 272.8/274.8 (M+H), retention time=0.97
min.
[0304] Step 2. tert-Butyl 6-bromopyridin-2-ylcarbamate (23.0 g, 84
mmol) was mixed with acetonitrile, (CH.sub.3CN, 281 mL), and NCS
(11.24 g, 84 mmol). The reaction mixture mixture was heated at
about 85.degree. C. for 3 hours, and an additional 5.5 g of NCS was
then added. Heating was continued at about 85.degree. C. for an
additional 3 hours, followed by addition of 5.5 g of NCS. All
starting materials were consumed after about 1 hour. Brine (50 mL)
was added and acetonitrile was evaporated under vacuum. The
residual aqueous solution was extracted three times with EtOAc. All
EtOAc layers were combined, dried over Na2SO4, filtered through a
fritted filter and concentrated under vacuum. The resulting residue
was purified on silica gel, eluting with 3% EtOAc in heptane to
afford the product as a colorless solid (14.6 g, 56.3%). LCMS
(m/z): 306.9/308.9/310.9 (M+H), retention time=1.14 min.
[0305] Step 3. A solution of tert-Butyl
6-bromo-5-chloropyridin-2-ylcarbamate (2.32 g, 7.54 mmol) in DMF
(25 mL) was mixed with sodium hydride (60% dispersion in mineral
oil. 513 mg, 12.8 mmol), and the resulting mixture reaction mixture
was stirred for 30 minutes at ambient temperature.
(2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl
4-methylbenzenesulfonate (3.15 g, 10.56 mmol), dissolved in 5 mL
DMF, was then added and the resulting mixture was stirred at about
25.degree. C. for 3 hours. The reaction mixture was partitioned
between water and EtOAc. The layers were separated and the EtOAc
layer was washed twice with water. The EtOAc layer was then dried
over sodium sulfate, filtered through a fritted filter and
concentrated under vacuum. The resulting residue was purified using
silica gel chromatography (0 to 30% EtOAc in heptane) to yield the
product as a colorless solid (2.16 g, 66%). LCMS (m/z):
432.9/434.9/436.9 (M+H), retention time=1.28 min.
[0306] Step 4. A mixture of tert-butyl
6-bromo-5-chloropyridin-2-yl((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl-
)carbamate (1.86 g, 4.29 mmol),
5-chloro-2-fluoropyridin-4-ylboronic acid (1.50 g, 8.58 mmol),
PdCl2(dppf)*DCM adduct (350 mg, 0.429 mmol), DME (15.6 mL) and 2 M
aqueous sodium carbonate solution (5.4 mL) were combined in a glass
bomb. The bomb was sealed and heated at about 98.degree. C. for 2
hours. The reaction mixture was cooled to ambient temperature and
then diluted with EtOAc. The diluted mixture was washed three times
with saturated aqueous NaHCO.sub.3 solution, dried over sodium
sulfate, filtered through a fritted filter and concentrated under
vacuum. Purification was done using silica gel chromatography (15%
EtOAc in heptane) to yield the product as a colorless solid (1.5 g,
72%). LCMS (m/z): 484.2/486.1 (M+H), retention time=1.33 min.
[0307] Step 5. tert-Butyl
3,5'-dichloro-2'-fluoro-2,4'-bipyridin-6-yl((2,2-dimethyltetrahydro-2H-py-
ran-4-yl)methyl)carbamate (8 mg, 0.017 mmol), DCM (1 mL) and TFA
(0.1 mL, 1.3 mmol) were combined in a 4 mL screw cap vial. The vial
was capped and the reaction mixture was stirred at ambient
temperature for 1 hour. The solvent was evaporated under vacuum and
the residual material was converted to the free base using sodium
bicarbonate. (5.8 mg, 91%). LCMS (m/z): 3484.2/386.1/388.2 (M+H),
retention time=1.07 min.
Synthesis of 2,3-difluoropyridin-4-ylboronic acid (Intermediate
W)
##STR00034##
[0309] A mixture of THF and hexanes (6 mL, 1:1 v:v), and
diisopropyl amine (0.681 mL, 4.78 mmol) was cooled to -78.degree.
C. BuLi (2.5 M in hexanes, 2.00 mL, 5.00 mmol) was added to the
cooled mixture, followed by addition of 2,3-difluoropyridine after
about 15 minutes. The mixture was stirred for 1 hour at -78.degree.
C. before being transferred to a 3 mL THF solution of triisopropyl
borate (1.11 mL, 4.78 mmol) at -78.degree. C. via a cannula. The
resulting solution was stirred at -78.degree. C. for 1 hour, slowly
warmed up to ambient temperature and then quenched with 2 M NaOH
solution (20 mL). The two layers were separated and the aqueous
phase was washed once with ether. The aqueous phase was then
acidified with HCl to pH 5 and extracted three times with EtOAc.
The organic layers were combined, dried over sodium sulfate and
concentrated to yield the product as a light yellow solid, which
was used in the next step without purification. LCMS (m/z): 159.9
(M+H), retention time=0.35 min.
Synthesis of
trans-N1-(1,3-dimethoxypropan-2-yl)cyclohexane-1,4-diamine
(Intermediate X)
##STR00035##
[0311] Step 1. To NaH (0.366 g, 9.16 mmol) in THF (12 mL) at
0.degree. C. was added 1,3-dimethoxy-2-propanol (1 g, 8.32 mmol) in
THF (8 mL) solution. The mixture was warmed to ambient temperature
and stirred for 0.5 hour. To this was added tosyl chloride (1.587
g, 8.32 mmol) in one portion. The resulting white cloudy mixture
then was stirred at ambient temperature for 16 hours. LC/MS showed
complete conversion to 1,3-dimethoxypropan-2-yl
4-methylbenzenesulfonate. The reaction mixture was poured into
water and extracted with EtOAc. The organic extracts were combined,
washed with brine, dried with sodium sulfate and concentrated in
vacuo to yield 2 g of a colorless oil. The crude mixture was
purified by Analogix system (silica gel column 80 g, gradient: 0
min, 100% n-heptane; 5-12 min, 20% EtOAc in Heptane; 12-15 min. 30%
EtOAc in Heptane and hold until 30 min). The pure fractions were
combined and concentrated in vacuo to yield 1.25 g of the tosylate
product 1,3-dimethoxypropan-2-yl 4-methylbenzenesulfonate as a
colorless oil, which solidified upon standing.
[0312] Step 2. To the tosylate obtained in Step 1 (0.8 g, 2.92
mmol) in DMSO (8 ml) was added 1,4-trans-cyclohexane diamine (0.999
g, 8.75 mmol). The resulting brown mixture was heated in a capped
vial to about 95.degree. C., with stirring, for 2 hours. The
reaction mixture was poured into 10% HCl in water (10 mL) at
0.degree. C. (ice cubes in HCl) and extracted with DCM (1.times.20
mL). The aqueous (light pink) was basified with 6N NaOH to a
pH>12 and extracted with DCM (2.times.20 mL). The organic
extracts were combined, dried with sodium sulfate and concentrated
in vacuo to yield compound "X" as a purple liquid. LC/MS showed
containing desired product (M+1=217, Rt=0.32 min, no UV absorption
at 214 nm wavelength). This was used in the next step without
further purification.
Synthesis of
4-((5'-chloro-2',5-difluoro-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2-
H-pyran-4-carbonitrile (Intermediate AA)
##STR00036##
[0313] Step 1: Synthesis of
4-((3,6-difluoropyridin-2-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitr-
ile
##STR00037##
[0315] To 2,3,6-Trifluoropyridine (0.6 g, 4.5 mmol) in DMSO (5 ml)
at room temperature was added
4-(aminomethyl)tetrahydro-2H-pyran-4-carbonitrile (Intermediate R,
1.01 g, 7.23 mmol) and triethylamine (1.57 ml, 11.24 mmol)
sequentially. The light brown mixture was heated at 105.degree. C.
in a sealed glass bomb for 18 hours. After cooled to room
temperature the reaction mixture was extracted with EtOAc (40 ml),
washed with saturated NaHCO.sub.3 solution and brine, dried over
sodium sulfate and concentrated in vacuo to give a light brown
liquid. This crude material was purified by silica gel
chromatography using a 12 g column, eluting with 5%-20%/ethyl
acetate in hexane to afford 550 mg (48.2% yield) of the desired
product. LCMS (m/z): 254.1 [M+H]+; retention time=0.743 min. 1H NMR
(400 MHz, CHLOROFORM-d) .delta. ppm 1.69-1.95 (m, 4H) 3.60-3.82 (m,
4H) 4.00 (ddd, J=12.13, 4.30, 1.96 Hz, 2H) 5.02 (br. S., 1H) 6.12
(td, J=5.58, 2.54 Hz, 1H) 7.19-7.33 (m, 1H).
Step 2: Synthesis of
4-((6-(benzyloxy)-3-fluoropyridin-2-yl-amino)methyl)tetrahydro-2H-pyran-4-
-carbonitrile
##STR00038##
[0317] Benzyl alcohol (352 mg, 3.26 mmol) was dissolved in
anhydrous DMF (2 ml) and placed under argon. This was then treated
with a 60% dispersion in oil of SODIUM HYDRIDE (78.7 mg, 3.26
mmol). This resultant suspension was then stirred at room
temperature for 15 min. At this time it was treated with a solution
of
4-((3,6-difluoropyridin-2-ylamino)methyl)tetrahydro-2H-pyran-4-carbonitri-
le (275 mg, 1.09 mmol) dissolved in anhydrous DMF (2 ml). Once the
addition was complete the reaction was stirred at 90.degree. C. for
5 hours. The reaction was allowed to cool to room temperature. It
was then poured into brine (20 ml). This was extracted with EtOAc
(3.times.15 ml). The combined extracts were washed with H.sub.2O
(3.times.10 ml) followed by brine (1.times.10 ml). The organic
layer was dried (Na.sub.2SO.sub.4), filtered, and the solvent
removed in vacuo to give the crude material which was purified
using the ISCO and a 12 g SiO.sub.2 column. Eluted using 100
hexanes to 30 EtOAc/70 hexanes over 20 min. 245 mg (66% yield) of
the desired product was obtained as a viscous liquid. LCMS (m/z):
342.1 [M+H]+; retention time=1.017 min.
Step 3: Synthesis of
4-((3-fluoro-6-hydroxypyridin-2-yl-amino)methyl)tetrahydro-2
H-pyran-4-carbonitrile
##STR00039##
[0319] A mixture of
4-((6-(benzyloxy)-3-fluoropyridin-2-ylamino)methyl)tetrahydro-2H-pyran-4--
carbonitrile (200 mg, 0.586 mmol), AMMONIUM FORMATE (111.3 mg,
1.758 mmol) and Pd--C (10%, wet, 25 mg) in methanol (4 ml) was
stirred at 70.degree. C. for 45 min and cooled. The mixture was
then filtered to remove Pd--C and inorganics, the filterate was
then concentrated and dried further via high vacuum to afford 141
mg (96% yield) of the crude product as a light pink solid. LCMS
(m/z): 252.1 [M+H]+; retention time=0.540 min.
Step 4: Synthesis
6-((4-cyanotetrahydro-2H-pyran-4-yl)methyl)-amino-5-fluoropyridin-2-yl
trifluoromethanesulfonate
##STR00040##
[0321] To a solution of
5-fluoro-6-((4-cyano-tetrahydro-2H-pyran-4-yl)methyl)aminopyridin-2-ol
(141 mg, 0.562 mmol) and TEA (0.782 ml, 5.60 mmol) in DCM (6 ml)
was added trifluoromethanesulfonic anhydride (0.142 ml, 0.842 mmol)
slowly at 0.degree. C. The mixture was stirred for 2 hours at
0.degree. C. and one hour at room temperature and poured carefully
into ice-cooled saturated aqueous NaHCO.sub.3 solution. The
separated aqueous layer was extracted with DCM (2.times.10 ml). The
combined organic layers were dried over Na.sub.2SO.sub.4, filtered
off and concentrated in vacuo. The residue was purified by column
chromatography [ISCO, SiO.sub.2, 12 g, 15 min, EtOAc/heptane=5/95
for 2 min, then EtOAc/heptane=5/95 to 40/60 for 2 min-17 min]. Pure
fractions were combined and concentrated in vacuo to give a
colorless oil (200 mg, 0.522 mmol, 93% yield) as the desired
product. LCMS (m/z): 384.0 [M+H]+; Rt=0.946 min.
Step 5: Synthesis of
4-((5'-chloro-2',5-difluoro-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2-
H-pyran-4-carbonitrile (Intermediate AA)
[0322] A mixture of
5-fluoro-6-((4-cyano-tetrahydro-2H-pyran-4-yl)methylamino)pyridin-2-yl
trifluoromethanesulfonate (200 mg, 0.522 mmol),
5-chloro-2-fluoropyridin-4-ylboronic acid (183.2 mg, 1.044 mmol),
PdCl.sub.2(dppf)-CH.sub.2Cl.sub.2 adduct (85.1 mg, 0.104 mmol), and
SODIUM CARBONATE (221.6 mg, 2.08 mmol, in 1 ml of water) in DME (3
ml) was de-gassed and heated at 110.degree. C. for 20 min in a
sealed microwave vial, cooled. The upper layer of mixture was
separated, the bottom one was extracted with ethyl acetates, the
organic layers were combined and concentrated to afford the crude
product, which was purified by ISCO (10 to 50% ethyl acetate in
heptane, 20 min) to afford 150 mg (79% yield) of the desired
product was an off-white solid. LCMS (m/z): 365.1 [M+H]+; retention
time=0.929 min.
Synthesis of
4-((5'-chloro-2'-fluoro-2,4'-bipyridin-6-yl-amino)methyl)tetrahydro-2H-py-
ran-4-carbonitrile (Intermediate AB)
##STR00041##
[0324] A mixture of
4-((6-bromopyridin-2-yl-amino)methyl)tetrahydro-2H-pyran-4-carbonitrile
(Intermediate S, 410 mg, 1.384 mmol),
5-chloro-2-fluoropyridin-4-ylboronic acid (362.2 mg, 2.07 mmol),
PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (113 mg, 0.14 mmol), DME
(5 Ml) and 2 M aqueous Na.sub.2CO.sub.2 (1.75 Ml, 3.5 mmol) was
sealed and stirred at 110.degree. C. for 20 min using microwave
reactor. After cooling to room temperature the mixture was
extracted with EtOAc (35 Ml), filtered and concentrated in vacuo.
The crude material was purified by column chromatography [silica
gel, 24 g, EtOAc/hexane=5/100 to 50/50] to provide
4-((5'-chloro-2'-fluoro-2,4'-bipyridin-6-ylamino)methyl)tetrahydr-
o-2H-pyran-4-carbonitrile (360 mg, 75% yield). LCMS (m/z): 347
[M+H]+; retention time=0.814 min.
Synthesis of
5'-chloro-2'-fluoro-N-((4-methoxytetrahydro-2H-pyran-4-yl)methyl)-2,4'-bi-
pyridin-6-amine (Intermediate AC)
##STR00042##
[0325] Step 1: Synthesis of tert-butyl
6-bromopyridin-2-yl((4-methoxytetrahydro-2H-pyran-4-yl)methyl)
carbamate
##STR00043##
[0327] To a solution of tert-butyl 6-bromo-pyridin-2-ylcarbamate
(136 mg, 0.50 mmol) in DMF (2 ml) under nitrogen was added NaH
(60%, 40 mg, 1.0 mmol) under stirring. The resultant mixture was
stirred at room temperature for one hour. A solution of
(4-methoxytetrahydro-2H-pyran-4-yl)methyl 4-methylbenzenesulfonate
(Intermediate O, 152 mg, 0.506 mmol) in DMF (1.5 ml) was then
added. The resulting mixture was then stirred at 85.degree. C. for
about 18 hours. The mixture was diluted with 30 ml of ethyl
acetate, washed with water (20 ml.times.3) and dried with sodium
sulfate. After concentration the residue was purified by silica gel
chromatography using a 12 g column, eluting with 5-20% ethyl
acetate in hexane to give the desired title compound as a viscous
oil (92 mg, 46% yield), which solidified upon standing overnight.
LCMS (m/z): 403.1 [M+H]+; Rt=1.026 min.
Step 2: Synthesis of tert-butyl
5'-chloro-2'-fluoro-2,4'-bipyridin-6-yl((4-methoxytetrahydro-2H-pyran-4-y-
l)methyl)carbamate
##STR00044##
[0329] A mixture of tert-butyl
6-bromo-pyridin-2-yl((4-methoxytetrahydro-2H-pyran-4-yl)methyl)carbamate
(50 mg, 0.125 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid
(43.7 mg, 0.249 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct
(15.2 mg, 0.019 mmol), DME (1.5 Ml) and 2M aqueous Na.sub.2CO.sub.2
(0.25 Ml, 0.5 mmol) was sealed and stirred at 100.degree. C. for 3
hours. After cooling to room temperature the mixture was diluted
with EtOAc (15 Ml), filtered and concentrated in vacuo. The crude
material was purified by column chromatography [silica gel, 12 g,
EtOAc/hexane=5/100 to 50/50] to provide tert-butyl
5'-chloro-2'-fluoro-2,4'-bipyridin-6-yl((4-methoxytetrahydro-2H-pyran-4-y-
l)methyl)carbamate (32 mg, 57% yield). LCMS (m/z): 452.2 [M+H]+;
retention time=1.068 min.
Step 3: Synthesis of
5'-chloro-2'-fluoro-N-((4-methoxytetrahydro-2H-pyran-4-yl)methyl)-2,4'-bi-
pyridin-6-amine (Intermediate AC)
[0330] A solution of tert-butyl
5'-chloro-2'-fluoro-2,4'-bipyridin-6-yl((4-methoxytetrahydro-2H-pyran-4-y-
l)methyl)carbamate (32 mg, 0.071 mmol) and TRIFLUOROACETIC ACID
(0.982 ml, 12.75 mmol) in DCM (2 ml) was stirred at room
temperature for 40 min. The mixture was then concentrated to afford
22 mg of the crude material which was used in the next step without
purification. LCMS (m/z): 352.2 [M+H]+; Rt=0.634 min.
Example 1a (Compound 1)
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridin-
e-2',6-diamine
##STR00045##
[0331] Step 1. Preparation of
trans-N1-(5-chloro-4-iodopyridin-2-yl)cyclohexane-1,4-diamine
[0332] A mixture of 5-chloro-2-fluoro-4-iodopyridine (1000 mg, 3.88
mmol), DMSO (7 ml), and trans-cyclohexane-1,4-diamine (2661 mg,
23.31 mmol) reaction mixture was stirred at about 85.degree. C. for
2 hours, followed by LCMS. The crude reaction mixture then was
mixed with 5 ml DMSO, filtered and purified by prep LC. After
lyapholization, 1.17 grams of the title compound was obtained as a
TFA salt. LCMS (m/z): 352.1 (MH+), retention time=0.50 min.
Step 2. Preparation of
trans-N1-(5'-chloro-6-fluoro-2,4'-bipyridin-2'-yl)cyclohexane-1,4-diamine
[0333] A mixture of
trans-N1-(5-chloro-4-iodopyridin-2-yl)cyclohexane-1,4-diamine (from
step 1 above, 300 mg, 0.853 mmol),
2-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(285 mg, 1.280 mmol), PdCl2(dppf).CH2Cl2 adduct (84 mg, 0.102
mmol), DME (4 ml), Ethanol (1 ml), and 2M sodium carbonate (1.706
ml, 3.41 mmol) reaction mixture was stirred at about 90.degree. C.
until done by LCMS. The reaction mixture was cooled, then diluted
with 25 ml of ethyl acetate and 10 ml of methanol, filtered, and
concentrated to yield a crude solid. The crude solid was dissolved
in DMSO, filtered and purified by prep LC. After lyapholization,
200 mg of the title compound was obtained as a TFA salt. LCMS
(m/z): 321.0 (MH+), retention time=0.48 min.
Step 3. Preparation of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine
[0334] To
trans-N1-(5'-chloro-6-fluoro-2,4'-bipyridin-2'-yl)cyclohexane-1,-
4-diamine (from Step 2 above, 200 mg, 0.623 mmol) was added DMSO (2
ml) and (3-fluorophenyl)methanamine (351 mg, 2.81 mmol). The crude
reaction mixture was stirred at 115.degree. C. until done, as
indicated by LCMS. The excess amine was removed under reduced
pressure. The resulting crude residue was dissolved in 2 ml of
DMSO, filtered, purified by prep LC and lyphilized to yield a TFA
salt. The TFA salt was free-based using 200 ml of ethyl acetate and
washed with saturated sodium bicarbonate 35 ml (1.times.), water
(2.times.), saturated brine (1.times.), dried over sodium sulfate,
filtered and concentrated to yield a solid. The solid was dissolved
in (1:1 ACN/water), filtered, and lyapholized to yield 80 mg of the
title compound as free-base. LCMS (m/z): 426.1 (MH+), retention
time=0.61 min.; 1H NMR (300 MHz, METHANOL-d4, 25.degree. C.)
1.21-1.40 (m, 4H) 1.89-2.00 (m, 2H) 2.07 (d, J=10.56 Hz, 2H)
2.69-2.79 (m, 1H) 3.55-3.64 (m, 1H) 4.57 (s, 2H) 6.53 (d, J=8.61
Hz, 1H) 6.59 (s, 1H) 6.80 (d, J=7.04 Hz, 1H) 6.90-6.97 (m, 1H) 7.09
(d, J=10.17 Hz, 1H) 7.14-7.20 (m, 1H) 7.25-7.34 (m, 1H) 7.48 (t,
J=7.83 Hz, 1H) 7.93 (s, 1H)
Example 1b (Compound 1)
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridin-
e-2',6-diamine
##STR00046##
[0335] Step 1. Preparation of
6-bromo-N-(3-fluorobenzyl)pyridin-2-amine
[0336] A mixture of 2,6-dibromopyridine (7.1 g, 30.0 mmol), NMP (16
ml), (3-fluorophenyl)methanamine (4.13 g, 33.0 mmol) and Hunig's
Base (5.76 ml, 33.0 mmol) was flushed with argon. The crude
reaction mixture was stirred at 115-120.degree. C. for about 168
hours. LC/MS was used to monitor the reaction. The crude mixture
was then cooled to room temperature, and then diluted with 250 ml
of ethyl acetate, washed with saturated sodium bicarbonate
(2.times.), water (2.times.), saturated. salt solution (1.times.),
dried over sodium sulfate, filtered, and concentrated under reduced
pressure to yield a residue. The residue was purified by silica gel
chromatography using a 120 g column, eluting from 0%-20% ethyl
acetate with hexane. The desired fractions were concentrated to
yield, 7.11 grams of the titled compound as a free base, which was
used in the next step without further purification. LCMS (m/z):
281.1/283.1 (MH+), retention time=1.03 min.
Step 2. Preparation of
5'-chloro-2'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine
[0337] A mixture of 6-bromo-N-(3-fluorobenzyl)pyridin-2-amine (2.0
g, 7.11 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid (1.996 g,
11.38 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (0.465 g,
0.569 mmol), DME (27 ml), and 2M sodium carbonate (9.25 ml, 18.50
mmol) reaction mixture was stirred at about 100.degree. C. for 3
hours. The crude mixture was cooled to room temperature, diluted
with 25 ml ethyl acetate and 20 ml methanol, filtered and
concentrated to yield crude residue. The crude residue was purified
by silica gel chromatography using a 120 g column, eluting from
0%-20% ethyl acetate with hexane. The desired fractions were
concentrated to constant mass, to yield 1.259 grams of titled
compound as free base, which was used in the next step without
further purification. LCMS (m/z): 332.2 (MH+), retention time=0.92
min.
Step 3. Preparation of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine
[0338] A mixture of
5'-chloro-2'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine (725
mg, 2.185 mmol) was added DMSO (7 ml),
trans-cyclohexane-1,4-diamine (1996 mg, 17.48 mmol) and TEA (0.609
ml, 4.37 mmol) was stirred at about 100.degree. C. for 20 hours.
The reaction was monitored by LC/MS. The crude reaction mixture was
cooled to room temperature, diluted with 3 ml DMSO, filtered, and
purified by prep HPLC. (there is a general HPLC conditions in the
general experimental session). The fractions were concentrated,
mixed with 500 ml ethyl acetate, and basified with saturated sodium
bicarbonate 120 ml. The ethyl acetate layer was separated, and the
basic water layer was extracted with 300 ml ethyl acetate. The
ethyl acetate layers were combined and washed with water
(3.times.), saturated salt solution (1.times.), dried with sodium
sulfate, filtered and concentrated to yield a solid. The solid was
dissolved in (1:1 ACN/water) filtered and lyapholized to yield 755
mg of the title compound as free-base. LCMS (m/z): 426.3 (MH+),
retention time=0.59 min.; 1H NMR (300 MHz, METHANOL-d4, 25.degree.
C.) .delta. ppm 1.10-1.43 (m, 4H) 1.90 (d, J=12.01 Hz, 2H) 2.01 (d,
J=12.01 Hz, 2H) 2.70-2.84 (m, 1H) 3.47-3.60 (m, 1H) 4.48 (s, 2H)
6.44 (d, J=8.50 Hz, 1H) 6.51 (s, 1H) 6.71 (d, J=7.33 Hz, 1H)
6.79-6.91 (m, 1H) 7.00 (d, J=9.96 Hz, 1H) 7.05-7.13 (m, 1H)
7.15-7.27 (m, 1H) 7.40 (t, J=7.77 Hz, 1H) 7.85 (s, 1H)
Example 2 (Compound 2)
[0339]
N2'-(trans-4-aminocyclohexyl)-N6-(cyclohexylmethyl)-2,4'-bipyridine-
-2',6-diamine
##STR00047##
Step 1. Preparation of
trans-N1-(4-bromopyridin-2-yl)cyclohexane-1,4-diamine
[0340] A mixture of 4-bromo-2-chloropyridine (1500 mg, 7.79 mmol),
DMSO (15 ml), and trans-cyclohexane-1,4-diamine (4450 mg, 39.0
mmol) was stirred at 100.degree. C. until the formation of the
product, as indicated by LCMS. The reaction mixture was cooled to
room temperature, filtered and purified by prep LC, and lyapholized
to yield 393 mg of the title compound as a TFA salt. LCMS (m/z):
270.2/272.2 (MH+), retention time=0.31 min.
Step 2. Preparation of
trans-N1-(6-fluoro-2,4'-bipyridin-2'-yl)cyclohexane-1,4-diamine
[0341] A mixture of
trans-N1-(4-bromopyridin-2-yl)cyclohexane-1,4-diamine (102 mg,
0.377 mmol),
2-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(80 mg, 0.359 mmol), PdCl2(dppf).CH.sub.2Cl.sub.2 adduct (29.3 mg,
0.036 mmol), DME (2 ml), Ethanol (0.2 ml), and 2M sodium carbonate
(0.717 ml, 1.435 mmol) reaction mixture was stirred at about
85.degree. C. until completion, as indicated by LCMS. The crude
mixture was cooled to room temperature, diluted with 5 ml of ethyl
acetate and 2 ml of methanol, filtered and concentrated to yield a
crude solid. The solid was dissolved in DMSO, refiltered, purified
by prep LC, and lyapholized to yield 64 mg of the title compound as
its TFA salt. LCMS (m/z): 287.2 (MH+), retention time=0.43 min.
Step 3. Preparation of
N2'-(trans-4-aminocyclohexyl)-N6-(cyclohexylmethyl)-2,4'-bipyridine-2',6--
diamine
[0342] A mixture of
trans-N1-(6-fluoro-2,4'-bipyridin-2'-yl)cyclohexane-1,4-diamine (15
mg, 0.052 mmol), DMSO (0.4 ml), and cyclohexylmethanamine (59.3 mg,
0.524 mmol) was heated at about 105.degree. C. for about 24 hours,
or until the product pormation was completed, as indicated by LCMS.
The excess amine was removed under reduced pressure to yield a
residue. The residue was mixed with 0.5 ml of DMSO, filtered and
purified by prep LC. After lyphilization, 11.3 mg of the title
compound was obtained as a TFA salt. LCMS (m/z): 380.3 (MH+),
retention time=0.61 min. 1H NMR (400 MHz, METHANOL-d4, 45.degree.
C.) .delta. ppm 0.97-1.11 (m, 2H) 1.17-1.36 (m, 3H) 1.49-1.72 (m,
6H) 1.71-1.80 (m, 2H) 1.84 (d, J=12.91 Hz, 2H) 2.11-2.28 (m, 4H)
3.13-3.25 (m, 1H) 3.28 (d, 2H, App.) 3.65-3.75 (m, 1H) 6.65 (d.
J=8.61 Hz, 1H) 7.16 (d, J=7.43 Hz, 1H) 7.43-7.48 (m, 1H) 7.52 (t,
J=7.83 Hz, 1H) 7.64 (s, 1H) 7.85 (d, J=7.04 Hz, 1H)
Example 3 (Compound 3)
trans-N1-(5-chloro-4-(6-(cyclohexylmethylamino)pyridin-2-yl)pyrimidin-2-yl-
)cyclohexane-1,4-diamine
##STR00048##
[0343] Step 1. Preparation of
2,5-dichloro-4-(6-fluoropyridin-2-yl)pyrimidine
[0344] A mixture of 2,4,5-trichloropyrimidine (49.3 mg, 0.269
mmol),
2-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(50 mg, 0.224 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct
(18.31 mg, 0.022 mmol), DME (0.7 ml), and 2M sodium carbonate
(0.247 ml, 0.493 mmol) reaction mixture was stirred at about
80.degree. C. until the reaction mixture was complete, as indicated
by LCMS. The reaction mixture was cooled, diluted with 5 ml of
ethyl acetate and 1 ml of methanol, filtered and concentrated to
yield a crude solid. The crude material was purified by silica gel
chromatography using a 12 g column, eluting from 0%-40% ethyl
acetate with hexane. The desired fractions were concentrated to
constant mass, to yield 39.5 mg of titled compound as a free base.
LCMS (m/z): 244.0 (MH+), retention time=0.89 min.
Step 2. Preparation of
trans-N1-(5-chloro-4-(6-fluoropyridin-2-yl)pyrimidin-2-yl)cyclohexane-1,4-
-diamine
[0345] A mixture of 2,5-dichloro-4-(6-fluoropyridin-2-yl)pyrimidine
(37 mg, 0.152 mmol), DMSO (1.5 ml) and
trans-cyclohexane-1,4-diamine (87 mg, 0.758 mmol) reaction mixture
was stirred at about 75.degree. C. for about 2 hours. The reaction
mixture was cooled, filter and purified by prep LC, and then
lyapholized to yield 39.5 mg of the title compound as a TFA salt.
LCMS (m/z): 322.2 (MH+), retention time=0.59 min.
Step 3. Preparation of trans-N1-(5-chloro-4-(6-(cyclohexylmethyl
amino)pyridin-2-yl)pyrimidin-2-yl)cyclohexane-1,4-diamine
[0346] A mixture of
trans-N1-(5-chloro-4-(6-fluoropyridin-2-yl)pyrimidin-2-yl)cyclohexane-1,4-
-diamine (12 mg, 0.037 mmol), cyclohexylmethanamine (42.2 mg, 0.373
mmol), and DMSO (0.35 ml) was stirred at about 105.degree. C. for
about 24 hours. The excess cyclohexylmethanamine was removed under
vacuum to yield a residue. The residue was mixed with 0.5 ml DMSO,
filtered, purified by prep HPLC and then lyapholized to yield 9.4
mg of the title compound as a TFA salt. LCMS (m/z): 415.3 (MH+),
retention time=0.67 min.; 1H NMR (400 MHz, METHANOL-d4, 45.degree.
C.) .delta. ppm 0.89-1.07 (m, 2H) 1.10-1.30 (m, 3H) 1.30-1.54 (m,
4H) 1.55-1.65 (m, 2H) 1.69 (d, J=12.91 Hz, 2H) 1.76 (d, J=12.91 Hz,
2H) 1.96-2.14 (m, 4H) 2.98-3.10 (m, 1H) 3.18 (d, J=6.65 Hz, 2H)
3.71-3.82 (m, 1H) 7.03 (d, J=9.00 Hz, 1H) 7.49 (br. s., 1H) 7.83
(t, J=8.22 Hz, 1H) 8.35 (s, 1H)
Example 4 (Compound 4)
(N2'-(trans-4-(aminomethyl)cyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'--
bipyridine-2',6-diamine
##STR00049##
[0347] Step 1. Preparation of tert-butyl
(trans-4-(5-chloro-4-iodopyridin-2-yl-amino)cyclohexyl)methylcarbamate
[0348] A mixture of 5-chloro-2-fluoro-4-iodopyridine (517 mg, 2.008
mmol), tert-butyl (trans-4-aminocyclohexyl)methylcarbamate (550 mg,
2.410 mmol), DMSO (2 ml) and TEA (0.336 ml, 2.410 mmol) reaction
mixture was stirred at about 95.degree. C. for about 26 hours. The
crude reaction mixture was cooled to room temperature, mixed with
125 ml ethyl acetate, washed with saturated sodium bicarbonate
(2.times.), water (3.times.), saturated salt solution (1.times.),
dried sodium sulfate, filtered and concentrated under reduced
pressure to yield a residue. The residue was purified by silica gel
chromatography using a 40 g column, eluting from 0%-35% ethyl
acetate with hexane. The desired fractions were concentrated to
constant mass, yielding 656 mg of titled compound as free base.
LCMS (m/z): 466.1 (MH+), retention time=0.93 min.
Step 2. Preparation of tert-butyl
(trans-4-(5'-chloro-6-fluoro-2,4'-bipyridin-2'-yl-amino)cyclohexyl)methyl-
carbamate
[0349] A mixture of tert-butyl
(trans-4-(5-chloro-4-iodopyridin-2-yl-amino)cyclohexyl)methylcarbamate
(510 mg, 1.095 mmol),
2-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(440 mg, 1.971 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (89
mg, 0.109 mmol), DME (7.5 ml), and 2M sodium carbonate (2.464 ml,
4.93 mmol) reaction mixture was stirred at about 100.degree. C. for
about 2 hours. The reaction mixture was cooled to room temperature,
mixed with 20 ml ethyl acetate, filtered and concentrated to yield
a crude solid. The crude solid was purified by silica gel
chromatography using 40 g column, eluting from 0%-45% ethyl acetate
with hexane. The desired fractions were concentrated to constant
mass, yielding 396 mg of titled compound as a free base. LCMS
(m/z): 435.2 (MH+), retention time=0.85 min.
Step 3. Preparation of
N-(trans-4-(aminomethyl)cyclohexyl)-5'-chloro-6-fluoro-2,4'-bipyridin-2'--
amine
[0350] A mixture of tert-butyl
(trans-4-(5'-chloro-6-fluoro-2,4'-bipyridin-2'-yl-amino)cyclohexyl)methyl-
carbamate (390 mg, 0.897 mmol), 4M HCl in Dioxane (5604 .mu.l,
22.42 mmol) reaction mixture was stirred at ambient temperature for
1 hr. The crude reaction mixture mixture was concentrated, and then
dried under high vacuum to a constant mass giving 335 mg of the
title compound as a HCL salt. LCMS (m/z): 335.1 (MH+), retention
time=0.51 min.
Step 4. Preparation of
N2'-(trans-4-(aminomethyl)cyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'--
bipyridine-2',6-diamine
[0351] A mixture of
N-(trans-4-(aminomethyl)cyclohexyl)-5'-chloro-6-fluoro-2,4'-bipyridin-2'--
amine (15 mg, 0.045 mmol), DMSO (0.35 ml), TEA (0.012 ml, 0.090
mmol) and (3-fluorophenyl)methanamine (50.5 mg, 0.403 mmol)
reaction mixture was flushed with argon and then stirred at about
105.degree. C. for about 40 hours. The excess
(3-fluorophenyl)methanamine was removed under reduced pressure to
yield a crude material, which was mixed with 0.5 mlDMSO, filtered,
purified by prep LC, and then lyapholized to yield 11.2 mg of the
title compound, as a TFA salt. LCMS (m/z): 440.2 (MH+), retention
time=0.62 min. 1H NMR (300 MHz, METHANOL-d4, 25.degree. C.) .delta.
ppm 1.11-1.28 (m, 2H) 1.28-1.47 (m, 2H) 1.67 (ddd, J=10.92, 7.40,
3.66 Hz, 1H) 1.92 (d, J=11.72 Hz, 2H) 2.14 (d, J=10.55 Hz, 2H) 2.83
(d, J=6.74 Hz, 2H) 3.57-3.69 (m, 1H) 4.63 (s, 2H) 6.84 (d, J=8.79
Hz, 1H) 6.90 (s, 1H) 6.94 (d, J=7.03 Hz, 1H) 6.96-7.03 (m, 1H) 7.10
(d, J=9.96 Hz, 1H) 7.18 (d, J=7.62 Hz, 1H) 7.29-7.39 (m, 1H)
7.69-7.77 (m, 1H) 8.01 (s, 1H)
Example 5 (Compound 5)
(5'-chloro-N6-(3-fluorobenzyl)-N2'-(piperidin-4-yl)-2,4'-bipyridine-2',6-d-
iamine
##STR00050##
[0352] Step 1. Preparation of tert-butyl
4-(5-chloro-4-iodopyridin-2-yl-amino)piperidine-1-carboxylate
[0353] A mixture of 5-chloro-2-fluoro-4-iodopyridine (517 mg, 2.008
mmol), tert-butyl 4-aminopiperidine-1-carboxylate (603 mg, 3.01
mmol), DMSO (2 ml) and TEA (0.420 ml, 3.01 mmol) reaction mixture
was stirred at 90.degree. C. for 18 hours. The reaction mixture was
cooled to room temperature, mixed with 150 ml of ethyl acetate,
washed with saturated sodium bicarbonate (2.times.), water
(3.times.), saturated salt solution (1.times.), dried sodium
sulfate, filtered and concentrated to yield a crude material, which
was purified by silica gel chromatography using a 40 g column,
eluting from 0%-40% ethyl acetate with hexane. The desired
fractions were concentrated to constant mass, giving 585 mg of the
title compound as free base. LCMS (m/z): 438.1 (MH+), retention
time=1.00 min.
Step 2. Preparation of tert-butyl
4-(5'-chloro-6-fluoro-2,4'-bipyridin-2'-yl-amino)piperidine-1-carboxylate
[0354] A mixture of tert-butyl
4-(5-chloro-4-iodopyridin-2-yl-amino)piperidine-1-carboxylate (468
mg, 1.069 mmol),
2-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(429 mg, 1.925 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (87
mg, 0.107 mmol), DME (7.5 ml), and 2M sodium carbonate (2.406 ml,
4.81 mmol) reaction mixture was stirred at 100.degree. C. for 2 hr.
The reaction mixture was cooled to room temperature, mixed with 20
ml of ethyl acetate, filtered and concentrated to yield a crude
material. The crude material was purified by silica gel
chromatography using a 40 g column, eluting from 0%-40% ethyl
acetate with hexane. The desired fractions were combined and
concentrated to constant mass, giving 360 mg of the title compound
as free base. LCMS (m/z): 407.2 (MH+), retention time=0.85 min.
Step 3. Preparation of tert-butyl
4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)piperidin-
e-1-carboxylate
[0355] A mixture of tert-butyl
4-(5'-chloro-6-fluoro-2,4'-bipyridin-2'-yl-amino)piperidine-1-carboxylate
(200 mg, 0.492 mmol), DMSO (2 ml), TEA (0.137 ml, 0.983 mmol) and
(3-fluorophenyl)methanamine (554 mg, 4.42 mmol) reaction mixture
was flushed with argon and stirred at 100.degree. C. for 40 hr, as
the reaction mixture progress was followed by LCMS. The reaction
mixture was cooled to room temperature, mixed with 150 ml of ethyl
acetate, washed with saturated sodium bicarbonate (2.times.), water
(3), saturated salt solution (1.times.), dried over sodium sulfate,
filtered and concentrated to yield a crude material, which was
purified by silica gel chromatography using a 12 g column, eluting
from 0%-35% ethyl acetate with hexane. The desired fractions were
collected and concentrated to constant mass, giving 225 mg of the
title compound as a free base. LCMS (m/z): 512.3 (MH.sup.+),
retention time=0.91 min.
Step 4. Preparation of
5'-chloro-N6-(3-fluorobenzyl)-N2'-(piperidin-4-yl)-2,4'-bipyridine-2',6-d-
iamine
[0356] A mixture of tert-butyl
4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)piperidin-
e-1-carboxylate (220 mg, 0.430 mmol), HCl 4M in Dioxane (7 mL, 28.0
mmol) was stirred at ambient temperature for 1 hr. The solvent was
evaporated under reduced pressure to yield a solid which was
further dried under high vacuum to yield 250 mg of the title
compound as a HCl salt. A portion of the title compound was
purified by prep LC, and then lyapholized to yield 19.0 mg of the
title compound as a TFA salt. LCMS (m/z): 412.2 (MH+), retention
time=0.60 min.; 1H NMR (300 MHz, METHANOL-d4, 25.degree. C.)
.delta. ppm 1.66-1.83 (m, 2H) 2.25 (dd, J=14.21, 3.08 Hz, 2H)
3.08-3.21 (m, 2H) 3.36-3.51 (m, 2H) 3.96-4.12 (m, 1H) 4.65 (s, 2H)
6.74 (s, 1H) 6.91 (s, 1H) 6.94 (s, 1H) 6.98-7.06 (m, 1H) 7.12 (d,
J=9.96 Hz, 1H) 7.19 (d, J=7.62 Hz, 1H) 7.31-7.43 (m, 1H) 7.77-7.85
(m, 1H) 8.09 (s, 1H)
Example 6 (Compound 6)
5'-chloro-N2'-(1-(ethylsulfonyl)piperidin-4-yl)-N6-(3-fluorobenzyl)-2,4'-b-
ipyridine-2',6-diamine
##STR00051##
[0357] Preparation of
5'-chloro-N2'-(1-(ethylsulfonyl)piperidin-4-yl)-N6-(3-fluoro
benzyl)-2,4'-bipyridine-2',6-diamine
[0358] A mixture of
5'-chloro-N6-(3-fluorobenzyl)-N2'-(piperidin-4-yl)-2,4'-bipyridine-2',6-d-
iamine (Example 6, 16 mg, 0.039 mmol), dichloromethane (0.5 ml),
and TEA (0.022 ml, 0.155 mmol) was cooled to 0.degree. C. This
cooled mixture was then diluted with a solution of 0.03 ml of
dichlormethane with ethanesulfonyl chloride (6.99 mg, 0.054 mmol).
The reaction mixture then was warmed to ambient temperature and
stirred for 1 hour, followed by LCMS. The reaction mixture solvent
was removed under reduced pressure, to yield a residue which was
dissolved in 0.75 ml DMSO, filtered, purified by prep LC and then
lyapholized to yield 9.9 mg of the title compound, as a TFA salt.
LCMS (m/z): 504.2 (MH+), retention time=0.77 min.; 1H NMR (300 MHz,
METHANOL-d4, 25.degree. C.) .delta. ppm 1.32 (t, J=7.33 Hz, 3H)
1.47-1.67 (m, 2H) 2.08 (d, J=10.84 Hz, 2H) 2.96-3.12 (m, 4H) 3.75
(d, J=12.89 Hz, 2H) 3.80-3.92 (m, 1H) 4.65 (s, 2H) 6.83 (s, 1H)
6.92 (d, J=9.08 Hz, 1H) 6.95 (d, J=7.62 Hz, 1H) 7.01 (t, J=8.64 Hz,
1H) 7.11 (d, J=9.96 Hz, 1H) 7.19 (d, J=7.62 Hz, 1H) 7.30-7.41 (m,
1H) 7.75-7.85 (m, 1H) 8.06 (s, 1H)
Example 7 (Compound 7)
N-(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)c-
yclohexyl)-2-(dimethylamino)acetamide
##STR00052##
[0359] Preparation of
N-(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)-
cyclohexyl)-2-(dimethylamino)acetamide
[0360] A mixture of 2-(dimethylamino)acetic acid (6.05 mg, 0.059
mmol), NMP (0.5 ml), Huenig's Base (0.023 ml, 0.132 mmol), and HATU
(24.55 mg, 0.065 mmol) was stirred at ambient temperature for 5
minutes, followed by addition of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine (Example 1) (12.5 mg, 0.029 mmol). The resulting
mixture was stirred at ambient temperature for 4 hours. The crude
reaction mixture was diluted with 0.25 ml of DMSO, filtered,
purified by prep LC and then lyapholized to yield 6.8 mg of the
title compound, as a TFA salt. LCMS (m/z): 511.3 (MH+), retention
time=0.62 min.; 1H NMR (300 MHz, METHANOL-d4, 25.degree. C.)
.delta. ppm 1.32-1.53 (m, 4H) 1.98-2.07 (m, 2H) 2.07-2.18 (m, 2H)
2.92 (s, 6H) 3.60-3.68 (m, 1H) 3.70-3.82 (m, 1H) 3.90 (s, 2H) 4.63
(s, 2H) 6.83 (d, J=8.79 Hz, 1H) 6.86 (s, 1H) 6.93 (d, J=7.03 Hz,
1H) 6.99 (s, 1H) 7.10 (d, J=9.67 Hz, 1H) 7.18 (d, J=7.62 Hz, 1H)
7.28-7.40 (m 1H) 7.68-7.77 (m, 1H) 8.01 (s, 1H)
Example 8 (Compound 8)
trans-4-(5'-chloro-6-(piperidin-4-yl-amino)-2,4'-bipyridin-2'-yl-amino)cyc-
lohexanol
##STR00053##
[0361] Step 1. Preparation of
trans-4-(5-chloro-4-iodopyridin-2-yl-amino)cyclohexanol
[0362] To 5-chloro-2-fluoro-4-iodopyridine (600 mg, 2.331 mmol) was
added DMSO (2.2 ml), trans-4-aminocyclohexanol (1074 mg, 9.32 mmol)
and TEA (0.390 ml, 2.80 mmol). The resulting reaction mixture was
stirred at 75.degree. C. for 24 hr, followed by LCMS. The reaction
mixture was cooled to room temperature, mixed with 150 ml of ethyl
acetate, washed with saturated sodium bicarbonate (1.times.), water
(1.times.), saturated salt solution (1.times.), dried over sodium
sulfate, filtered and concentrated to yield a crude material. The
crude material was purified by silica gel chromatography using a 40
g column eluting from 15%-75% ethyl acetate with hexane. The
desired fractions were combined and concentrated to constant mass,
giving 750 mg of the title compound as free base, which was used in
the next step without further purification. LCMS (m/z): 353.0
(MH+), retention time=0.56 min.
Step 2. Preparation of
trans-4-(5'-chloro-6-fluoro-2,4'-bipyridin-2'-yl-amino)cyclohexanol
[0363] A mixture of
trans-4-(5-chloro-4-iodopyridin-2-yl-amino)cyclohexanol (575 mg,
1.631 mmol),
2-fluoro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine
(655 mg, 2.94 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (133
mg, 0.163 mmol), DME (15 ml), and t 2M sodium carbonate (4.48 ml,
8.97 mmol) reaction mixture was stirred at 95.degree. C. for 2 hr,
followed by LCMS. The reaction mixture was cooled to room
temperature, mixed with 20 ml of ethyl acetate, 15 ml of methanol,
filtered and concentrated to yield a crude product. The crude was
purified by silica gel chromatography using a 40 g column, eluting
from 35%-85% ethyl acetate with hexane. The desired fractions were
combined and concentrated to constant mass, giving 440 mg of titled
compound as free base. LCMS (m/z): 322.2 (MH+), retention time=0.53
min.
Step 3. Preparation of
trans-4-(5'-chloro-6-(piperidin-4-yl-amino)-2,4'-bipyridin-2'-yl-amino)cy-
clohexanol
[0364] A mixture of
trans-4-(5'-chloro-6-fluoro-2,4'-bipyridin-2'-yl-amino)cyclohexanol
(15.5 mg, 0.048 mmol), DMSO (0.4 ml), and tert-butyl
4-aminopiperidine-1-carboxylate (48.2 mg, 0.241 mmol) reaction
mixture was stirred at 105.degree. C. for 40 hr. LCMS indicated
formation of the intermediate tert-butyl
4-(5'-chloro-2'-(trans-4-hydroxycyclohexylamino)-2,4'-bipyridin-6-yl-amin-
o)piperidine-1-carboxylate (LCMS (m/z): 502.4 (MH+), retention
time=0.70 min.). The Boc protecting group was removed from the
intermediate by adding HCL 6M aq (140 .mu.l, 0.840 mmol) to the
crude reaction mixture, followed by stirring the mixture at
90.degree. C. for 45 minutes. The reaction mixture was cooled, 0.5
ml of DMSO was added, filtered and purified by prep LC.
Lyapholization of the material yielded 9.8 mg of the title
compound, as a TFA salt. LCMS (m/z): 402.3 (MH+), retention
time=0.41 min.; 1H NMR (300 MHz, METHANOL-d4, 25.degree. C.)
.delta. ppm 1.32-1.52 (m, 4H) 1.71-1.87 (m, 2H) 1.96-2.12 (m, 4H)
2.27 (dd, J=14.21, 3.37 Hz, 2H) 3.06-3.18 (m, 2H) 3.39-3.50 (m, 2H)
3.54-3.68 (m, 2H) 4.05-4.17 (m, 1H) 6.72 (d, J=8.50 Hz, 1H) 6.90
(d, J=7.33 Hz, 1H) 7.00 (s, 1H) 7.56-7.64 (m, 1H) 8.01 (s, 1H)
Example 9 (Compound 9)
N-(trans-4-(aminomethyl)cyclohexyl)-5'-chloro-6-(3-fluorobenzyloxy)-2,4'-b-
ipyridin-2'-amine
##STR00054##
[0365] Step 1. Preparation of
2-bromo-6-(3-fluorobenzyloxy)pyridine
[0366] To 2-bromo-6-fluoropyridine (176 mg, 1.000 mmol) was added
DMF (1.5 ml) and (3-fluorophenyl)methanol (139 mg, 1.100 mmol) and
cesium carbonate (391 mg, 1.200 mmol), and the resulting mixture
reaction mixture was stirred at 95.degree. C. for 6 hr, as the
progress of the reaction mixture was followed by LCMS. The reaction
mixture was cooled to room temperature, diluted with 120 ml of
ethyl acetate, washed with saturated sodium bicarbonate (1.times.),
water (1.times.), saturated salt solution (1.times.), dried over
sodium sulfate, filtered and concentrated to yield a crude product
which was purified by silica gel chromatography using a 12 g column
eluting from 0%-20% ethyl acetate with hexane. The desired
fractions were combined and concentrated to constant mass, giving
156 mg of the title compound as a free base. LCMS (m/z):
282.0/284.0 (MH+), retention time=1.19 min.
Step 2. Preparation of
5'-chloro-2'-fluoro-6-(3-fluorobenzyloxy)-2,4'-bipyridine
[0367] A mixture of 2-bromo-6-(3-fluorobenzyloxy)pyridine (145 mg,
0.514 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid (144 mg,
0.822 mmol), PalladiumTetrakis (71.3 mg, 0.062 mmol), DME (3 ml),
and t 2M sodium carbonate (1.028 ml, 2.056 mmol) was reaction
mixture was stirred at 100.degree. C. for 3 hr, followed by LCMS.
The reaction mixture was cooled, diluted with 10 ml of ethyl
acetate, filtered and concentrated to yield a crude product, which
was purified by silica gel chromatography using a 12 g column
eluting from 0%-20% ethyl acetate with hexane. The desired
fractions were concentrated to constant mass, giving 100 mg of
titled compound as a free base. LCMS (m/z): 333.1 (MH+), retention
time=1.26 min.
Step 3. Preparation of
N-(trans-4-(aminomethyl)cyclohexyl)-5'-chloro-6-(3-fluorobenzyloxy)-2,4'--
bipyridin-2'-amine
[0368] To 5'-chloro-2'-fluoro-6-(3-fluorobenzyloxy)-2,4'-bipyridine
(30 mg, 0.090 mmol) was added DMSO (0.8 ml). TEA (0.025 ml, 0.180
mmol), and tert-butyl (trans-4-aminocyclohexyl)methylcarbamate
(41.2 mg, 0.180 mmol). The reaction mixture was flushed with argon
and stirred at 100-105.degree. C. for 40 hr. Formation of the
intermediate product tert-butyl
(trans-4-(5'-chloro-6-(3-fluorobenzyloxy)-2,4'-bipyridin-2'-yl-amino)cycl-
ohexyl)methylcarbamate was indicated by LCMS. (LCMS (m/z): 541.4
(MH+), retention time=1.05 min.). The solvent DMSO was removed
under reduce pressure. The Boc group was removed from the
intermediate by adding 4M HCl in Dioxane (1.5 ml, 6.00 mmol),
followed with stirring at ambient temperature for 90 minutes. The
solvent was removed under reduced pressure. The crude product was
dissolved in 1.0 ml of DMSO with 0.075 ml of water, filtered and
purified by prep LC. After lyphilization, 28.3 mg of the title
compound was obtained as a TFA salt. LCMS (m/z): 441.3 (MH+),
retention time=0.76 min.; 1H NMR (300 MHz, METHANOL-d4, 25.degree.
C.) .delta. ppm 1.12-1.29 (m, 2H) 1.29-1.47 (m, 2H) 1.60-1.76 (m,
J=14.76, 7.51, 3.66, 3.66 Hz, 1H) 1.92 (d, J=12.60 Hz, 2H) 2.16 (d,
J=10.55 Hz, 2H) 2.84 (d, J=6.74 Hz, 2H) 3.58-3.71 (m, 1H) 5.43 (s,
2H) 6.92-6.99 (m, 2H) 6.99-7.08 (m, 1H) 7.18 (d, J=9.67 Hz, 1H)
7.25 (d, J=7.62 Hz, 1H) 7.30-7.42 (m, 2H) 7.83 (t, J=7.77 Hz, 1H)
8.01 (s, 1H)
Example 10 (Compound 10)
trans-N1-benzyl-N4-(4-(6-(3-fluorobenzylamino)pyrazin-2-yl)pyridin-2-yl)cy-
clohexane-1,4-diamine
##STR00055##
[0369] Step 1. Preparation of
6-chloro-N-(3-fluorobenzyl)pyrazin-2-amine To 2,6-dichloropyrazine
(175 mg, 1.175 mmol) was added DMSO (1.5 ml), TEA (0.196 ml, 1.410
mmol) and (3-fluorophenyl)methanamine (368 mg, 2.94 mmol)l. The
reaction mixture then was stirred at 90.degree. C. until completion
as indicated by LCMS, about 1 hour. To the reaction mixture was
added 3 ml of DMSO, filtered and the residue was purified by prep
LC. After lyphilization, 160 mg of the title compound was obtained
as a TFA. LCMS (m/z): 238.1 (MH+), retention time=0.96 min.
Step 2. Preparation of
N-(3-fluorobenzyl)-6-(2-fluoropyridin-4-yl)pyrazin-2-amine
[0370] To 6-chloro-N-(3-fluorobenzyl)pyrazin-2-amine (140 mg, 0.589
mmol) was added 2-fluoropyridin-4-ylboronic acid (125 mg, 0.884
mmol), PalladiumTetrakis (82 mg, 0.071 mmol), DME (3.3 ml), and 2M
sodium carbonate (1.031 ml, 2.062 mmol). The resulting reaction
mixture was stirred at 110.degree. C. until completion as indicated
by LCMS, about 3 hours. The reaction mixture was cooled to room
temperature, diluted with 20 ml of ethyl acetate, filtered and
concentrated to yield a crude solid. The solid was dissolved in
DMSO, filtered and purified by prep LC. After lyphilization, 72 mg
of the title compound was botained as a TFA salt. LCMS (m/z): 299.1
(MH+), retention time=0.89 min.
Step 3. Preparation of
trans-N1-(4-(6-(3-fluorobenzylamino)pyrazin-2-yl)pyridin-2-yl)cyclohexane-
-1,4-diamine
[0371] To
N-(3-fluorobenzyl)-6-(2-fluoropyridin-4-yl)pyrazin-2-amine (30 mg,
0.101 mmol) was added DMSO (0.6 ml) and
trans-cyclohexane-1,4-diamine (115 mg, 1.006 mmol). The reaction
mixture then was stirred at 105.degree. C. until completion as
indicated by LCMS, about 40 hours. To the crude reaction mixture,
after cooling to room temperature mixture was added 0.75 ml of
DMSO, the resulting mixture filtered and purified by prep LC. After
lyphilization, 34 mg of the title compound was obtained as a TFA
salt. LCMS (m/z): 393.2 (MH+), retention time=0.54 min.
Step 4. Preparation of
trans-N1-benzyl-N4-(4-(6-(3-fluorobenzylamino)
pyrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
[0372] To trans-N1-(4-(6-(3-fluorobenzyl
amino)pyrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine (19 mg,
0.048 mmol) was added NMP (0.6 ml), acetic acid (0.042 ml, 0.726
mmol) and benzaldehyde (10.27 mg, 0.097 mmol). The resulting
reaction mixture was stirred overnight at ambient temperature. To
the stirred reaction mixture was added sodium triacetoxyborohydride
(41.0 mg, 0.194 mmol) and the resulting mixture was stirred
overnight (24 hours) at ambient temperature. To the reaction
mixture then was added additional sodium triacetoxyborohydride
(21.0 mg, 0.099 mmol) and the resulting mixture was stirred for an
additional 2 more hours. To the crude mixture then was added 0.8 ml
of DMSO, filtered and purified by prep LC. After lyphilization, 7.0
mg of the title compound was obtained as a TFA salt. LCMS (m/z):
483.2 (MH+), retention time=0.65 min.; 1H NMR (300 MHz,
METHANOL-d4, 25.degree. C.) .delta. ppm 1.35-1.51 (m, 2H) 1.51-1.69
(m, 2H) 2.04-2.36 (m, 4H) 3.08-3.18 (m, 1H) 3.56-3.70 (m, 1H) 4.16
(s, 2H) 4.60 (s, 2H) 6.82-6.93 (m, 1H) 7.03 (d, J=9.67 Hz, 1H) 7.11
(d, J=7.62 Hz, 1H) 7.19-7.29 (m, 1H) 7.32 (d, J=6.74 Hz, 1H)
7.35-7.46 (m, 5H) 7.54 (s, 1H) 7.80 (d, J=6.74 Hz, 1H) 7.97 (s, 1H)
8.25 (s, 1H)
Example 11 (Compound 11)
N2'-(trans-4-aminocyclohexyl)-N6-(3-fluorobenzyl)-4-(trifluoromethyl)-2,4'-
-bipyridine-2',6-diamine
##STR00056##
[0373] Step 1. Preparation of
6-chloro-N-(3-fluorobenzyl)-4-(trifluoromethyl)pyridin-2-amine
[0374] To 2,6-dichloro-4-(trifluoromethyl)pyridine (250 mg, 1.157
mmol) was added DMSO (2 ml), TEA (0.194 ml, 1.389 mmol), and
(3-fluorophenyl)methanamine (290 mg, 2.315 mmol). The reaction
mixture was stirred at 90.degree. C. until completion as indicated
by LCMS, about 1 hour. To the crude reaction mixture was added 1.5
ml of DMSO, filtered and purified by prep LC. After lyphilization,
158 mg of the title compound was obtained as a TFA salt. LCMS
(m/z): 305.1 (MH+), rt=1.21 min.
Step 2. Preparation of
2'-fluoro-N-(3-fluorobenzyl)-4-(trifluoromethyl)-2,4'-bipyridin-6-amine
[0375] To
6-chloro-N-(3-fluorobenzyl)-4-(trifluoromethyl)pyridin-2-amine (70
mg, 0.230 mmol) was added 2-fluoropyridin-4-ylboronic acid (58.3
mg, 0.414 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (22.52
mg, 0.028 mmol), DME (1.2 ml), and 2M sodium carbonate (0.460 ml,
0.919 mmol). The resulting reaction mixture was stirred at
105.degree. C. until completion as indicated by LCMS, about 6
hours. The reaction mixture was cooled, 15 ml of ethyl acetate and
5 ml of methanol was added, filtered and concentrated to yield a
crude solid. The solid was purified by prep LC. The product was
free-based using 200 ml of ethyl acetate and washed with saturated
sodium bicarbonate (1.times.), water (2.times.), saturated salt
solution (1.times.), dried sodium sulfate, filtered and
concentrated to a constant mass, yielding 35 mg of titled compound
as free base. LCMS (m/z): 366.2 (MH+), retention time=1.20 min.
Step 3. preparation of
N2'-(trans-4-aminocyclohexyl)-N6-(3-fluorobenzyl)-4-(trifluoromethyl)-2,4-
'-bipyridine-2',6-diamine
[0376] To
2'-fluoro-N-(3-fluorobenzyl)-4-(trifluoromethyl)-2,4'-bipyridin--
6-amine (34 mg, 0.093 mmol) was added DMSO (1.7 ml) and
trans-cyclohexane-1,4-diamine (159 mg, 1.396 mmol). The resulting
reaction mixture was stirred at 105.degree. C. until completion as
indicated by LCMS, about 40 hours. To the crude reaction mixture
was added 0.75 ml of DMSO, filtered and purified by prep LC. After
lyphilization, 28.1 mg of the title compound was obtained as a TFA
salt. LCMS (m/z): 460.3 (MH+), retention time=0.72 min.; 1H NMR
(300 MHz, METHANOL-d4, 25.degree. C.) .delta. ppm 1.40-1.72 (m, 4H)
2.18 (t, J=13.77 Hz, 4H) 3.11-3.24 (m, 1H) 3.62-3.76 (m, 1H) 4.72
(s, 2H) 6.95 (s, 1H) 7.12 (d, J=9.96 Hz, 1H) 7.17-7.24 (m, 1H)
7.27-7.37 (m, 1H) 7.40 (s, 1H) 7.42-7.48 (m, 1H) 7.69 (s, 1H) 7.87
(d, J=6.74 Hz, 1H)
Example 12 (Compound 12)
2-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N.sup.6-(3-fluorobenzyl)-2,-
4'-bipyridine-2',6-diamine
##STR00057##
[0377] Step 1. Preparation of
3,6-difluoro-N-(3-fluorobenzyl)pyridine-2-amine
[0378] 2,3,6-trifluoropyridine (1.07 mL, 1.5 g, 11.27 mmol),
3-fluorobenzylamine (3.18 mL, 3.53 g, 28.2 mmol), and triethylamine
(4.71 mL, 3.42 g, 33.8 mmol) were dissolved in NMP (39 mL) to form
a mixture This mixture reaction mixture was stirred at 100.degree.
C. for 1 hr. The reaction mixture was then extracted with EtOAc
(3.times.75 mL). The combined extracts were washed with H.sub.2O
(4.times.75 mL) followed by brine (1.times.75 mL). The organic
layer was dried (Na.sub.2SO.sub.4), filtered, and the solvent
removed in vacuo. The resulting residue was subjected to silica gel
column chromatography. Elution using 100 hexanes to 30 EtOAc/70
hexanes yielded 2.63 g (98%) of
3,6-difluoro-N-(3-fluorobenzyl)pyridine-2-amine. LCMS (m/z): 239.1
(MH.sup.+), retention time=1.01 min.
Step 2. Preparation of
3-fluoro-N-(3-fluorobenzyl)-6-methoxypyridin-2-amine
[0379] 3,6-difluoro-N-(3-fluorobenzyl)pyridine-2-amine (0.5209 g,
2.19 mmol), was dissolved in anhydrous MeOH (6.6 mL) and placed
under argon. This mixture then was treated with sodium methoxide
(0.500 mL, 0.473 g, 2.19 mmol, 25% in MeOH) by slow addition. The
resulting mixture was then heated in the microwave at 150.degree.
C. for four 30 min. The reaction mixture was then poured into brine
(25 mL). This mixture was extracted with EtOAc (3.times.25 mL), the
combined extracts were washed with brine (1.times.25 mL) and dried
(Na.sub.2SO.sub.4). After filtration the solvent removed in vacuo.
The resulting residue was subjected to silica gel column
chromatography. Elution using 100 hexanes to 25 EtOAc/75 hexanes
afforded 0.3408 g (62%) of
3-fluoro-N-(3-fluorobenzyl)-6-methoxypyridin-2-amine. LCMS (m/z):
251.1 (MH.sup.+), retention time=1.07 min.
Step 3. Preparation of
5-fluoro-6-(3-fluorobenzylamino)pyridine-2-ol
[0380] 3-fluoro-N-(3-fluorobenzyl)-6-methoxypyridin-2-amine (0.100
g, 0.400 mmol) was dissolved in anhydrous CH.sub.3CN (1.6 mL). This
mixture was treated with sodium iodide (0.301 g, 2.01 mmol)
followed by trimethylsilylchloride (0.257 mL, 0.218 g, 2.01 mmol).
The resulting reaction mixture was then heated at reflux for 2 hr.
The reaction mixture was then treated with MeOH (1 ml), and the
resulting mixture was stirred at ambient temperature for 2 hr, and
then concentrated in vacuo. The resulting residue was dissolved in
EtOAc (25 ml) and partitioned with H.sub.2O (25 ml). The H.sub.2O
layer was extracted with EtOAc (2.times.25 ml). The organic layers
were combined and washed with brine (1.times.25 ml). The organic
layer was dried (Na.sub.2SO.sub.4), filtered, and the solvent
removed in vacuo. The resulting residue was subjected to silica gel
column chromatography. Elution using 10 EtOAc/90 hexanes to 60
EtOAc/40 hexanes gave 0.060 g (64%) of
5-fluoro-6-(3-fluorobenzylamino)pyridine-2-ol. LCMS (m/z): 237.2
(MH.sup.+), retention time=0.74 min.
Step 4. Preparation of
5-fluoro-6-(3-fluorobenzylamino)pyridine-2-yl
trifluoromethanesulfonate
[0381] 5-fluoro-6-(3-fluorobenzylamino)pyridine-2-ol (0.060 g,
0.254 mmol) was dissolved in anhydrous CH.sub.2Cl.sub.2 (2.0 mL)
and placed under argon. The solution was cooled to 0.degree. C. in
an ice bath. It was then treated with triethylamine (0.096 mL,
0.070 g, 0.691 mmol) followed by dropwise addition of
trifluoromethanesulfonic anhydride (0.058 mL, 0.096 g, 0.340 mmol).
Once the addition was complete, the reaction mixture was stirred at
0.degree. C. for 2 hr. The reaction mixture was then poured into
saturated NaHCO.sub.3 (25 mL). This mixture was extracted with
EtOAc (2.times.25 mL). The combined extracts were washed with brine
(1.times.25 mL), dried (Na.sub.2SO.sub.4), filtered, and the
solvent removed in vacuo. The resulting residue was subjected to
silica gel column chromatography. Elution using 5 EtOAc/95 hexanes
to 60 EtOAc/40 hexanes yielded 0.081 g (87%) of
5-fluoro-6-(3-fluorobenzylamino)pyridine-2-yl
trifluoromethanesulfonate. LCMS (m/z): 369.1 (MH.sup.+), retention
time=1.15 min.
Step 5. Preparation of
5'-chloro-2',5-difluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine
[0382] 5-fluoro-6-(3-fluorobenzylamino)pyridine-2-yl
trifluoromethanesulfonate (0.0811 g, 0.220 mmol),
5-chloro-2-fluoropyridin-4-ylboronic acid (0.116 g, 0.661 mmol),
and sodium carbonate (0.286 mL, 0.573 mmol, 2M in H.sub.2O) were
dissolved in DME (2 mL). The solution was then degassed by sparging
with argon for 5 min. It was then treated with PdCl.sub.2(dppf)
CH.sub.2Cl.sub.2 adduct (0.036 g, 0.044 mmol). The reaction mixture
was then heated in the microwave at 120.degree. C. for 10 min. The
reaction mixture was then filtered through a pad of Celite. The
filtrate was concentrated in vacuo. The resulting residue was
subjected to silica gel column chromatography. Elution using 5
EtOAc/95 hexanes to 60 EtOAc/40 hexanes yielded 0.044 g (57%) of
5'-chloro-2',5-difluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amin-
e. LCMS (m/z): 350.0 (MH.sup.+), retention time=1.16 min.
Step 6. Preparation of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-(3-fluorobenzyl)-2,4'-
-bipyridine-2',6-diamine
[0383]
5'-chloro-2',5-difluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine
(0.022 g, 0.063 mmol) was dissolved in anhydrous DMSO (0.93 mL) and
charged to a microwave vial, and then treated with
trans-cyclohexane-1,4-diamine (0.072 g, 0.629 mmol). The reaction
mixture then was heated at 100.degree. C. for 18 hr. The material
was purified by preparative reverse-phase HPLC to yield 0.0151 g
(44%) of
N.sup.2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N.sup.6-(3-fluorobe-
nzyl)-2,4'-bipyridine-2',6-diamine as the TFA salt. LCMS (m/z):
444.2 (MH.sup.+), retention time=0.7 min.
Example 13 (Compound 13)
2'-((1r,4r)-4-amonocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'-
-bipyridine-5-carbonitrile
##STR00058##
[0384] Step 1. Preparation of
6-chloro-2-(3-fluorobenzylamino)nicotinonitrile
[0385] 2,6-dichloronicotinonitrile (0.500 g, 2.89 mmol),
(4-fluorophenyl)methanamine (0.816 mL, 0.904 g, 7.23 mmol), and
triethylamine (1.21 mL, 0.877 g, 8.67 mmol) were all mixed in NMP
(10 mL). The resulting solution then was heated at 50.degree. C.
for 18 hr. The reaction mixture was then poured in H.sub.2O (25
mL), and extracted with EtOAc (3.times.25 mL). The combined
extracts were washed with H.sub.2O (4.times.25 mL), and brine
(1.times.25 mL). The organic layer was separated and dried
(Na.sub.2SO.sub.4), filtered, and the solvent removed in vacuo. The
resulting residue was purified using silica gel column
chromatography. Elution using 1 EtOAc/3 hexanes to 3 EtOAc/1
hexanes afforded 0.6024 g (80%) of
6-chloro-2-(3-fluorobenzylamino)nicotinonitrile. LCMS (m/z): 350.0
(MH.sup.+), retention time=0.96 min. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 4.58 (d, J=5.86 Hz, 2H) 5.48 (br. s., 1H) 6.30
(d, J=8.50 Hz, 1H) 6.96-7.06 (m, 2H) 7.10 (d, J=7.62 Hz, 1H)
7.28-7.38 (m, 1H) 7.58 (d, J=8.79 Hz, 1H).
Step 2. Preparation of
5'-chloro-2'-fluoro-6-(3-fluorobenzylamino)-2,4'-bipyridine-5-carbonitril-
e
[0386] 6-chloro-2-(3-fluorobenzylamino)nicotinonitrile (0.602 g,
2.30 mmol). 5-chloro-2-fluoropyridin-4-ylboronic acid (1.21 g, 6.91
mmol), and sodium carbonate (2.99 mL, 5.99 mmol, 2M in H.sub.2O)
were dissolved in DME (10.5 mL). The resulting solution was then
degassed by sparging with argon for 5 min. It was then treated with
PdCl.sub.2(dppf) CH.sub.2Cl.sub.2 adduct (0.376 g, 0.460 mmol). The
resulting reaction mixture was heated in the microwave at
120.degree. C. for 10 min. It was then filtered through a pad of
Celite. The filtrate was concentrated in vacuo. The resulting
residue was subjected to silica gel column chromatography. Elution
using 5 EtOAc/95 hexanes to 50 EtOAc/50 hexanes yielded 0.2689 g
(33%) of
5'-chloro-2'-fluoro-6-(3-fluorobenzylamino)-2,4'-bipyridine-5-carbonitril-
e. LCMS (m/z): 357.2 (MH.sup.+), retention time=1.02 min.
Step 3. Preparation of
2'-((1r,4r)-4-amonocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4-
'-bipyridine-5-carbonitrile
[0387]
5'-chloro-2'-fluoro-6-(3-fluorobenzylamino)-2,4'-bipyridine-5-carbo-
nitrile (0.2689 g, 0.754 mmol) was dissolved in anhydrous DMSO
(11.0 mL) and charged to a microwave vial. This mixture was treated
with trans-cyclohexane-1,4-diamine (0.861 g, 7.54 mmol), and the
reaction mixture was then heated at 100.degree. C. for 5 hr. The
reaction mixture mixture was cooled to ambient temperature, and the
material was purified by preparative reverse-phase HPLC and
freebased to yield 0.2539 g (75%) of
2'-((1r,4r)-4-amonocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)--
2,4'-bipyridine-5-carbonitrile. LCMS (m/z): 451.2 (MH.sup.+),
retention time=0.67 min.
Example 14 (Compound 14)
2'-((1r,4r)-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'-
-bipyridine-5-carboxamide
##STR00059##
[0388] Step 1. Preparation of
2'-((1r,4r)-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4-
'-bipyridine-5-carboxamide
[0389] 2'-((1r,4r)-4-amonocyclohexyl
amino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridine-5-carbonitrile
(0.028 g, 0.055 mmol) was dissolved in DMSO (0.5 mL), and the
solution was cooled to 0.degree. C. in an ice bath. The cooled
solution was treated with potassium carbonate (0.0011 g, 0.0078
mmol) followed by hydrogen peroxide (0.007 mL, 0.0069 mmol). The
ice bath was removed and the reaction mixture was stirred at
ambient temperature for 2 hr. More of the reagents in the same
amounts were added and the reaction mixture was heated to
50.degree. C. for 16 hr. This procedure was repeated and the
reaction mixture was heated at 65.degree. C. for an additional 4
hr. The reaction mixture was diluted with brine (10 mL), extracted
with EtOAc (3.times.10 m L), the combined extracts were washed with
brine (1.times.10 mL) and dried (Na.sub.2SO.sub.4), filtered and
concentrated in vacuo. The material was purified by preparative
reverse-phase HPLC to afford 0.0042 g (13.degree. %) of
2'-((1r,4r)-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4-
'-bipyridine-5-carboxamide as the TFA salt. LCMS (m/z): 469.1
(MH.sup.+), retention time=0.56 min.
Example 15 (Compound 15)
[0390]
2'-(trans-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-
-2,4'-bipyridine-4-carbonitrile
##STR00060##
Step 1. Preparation of
2-chloro-6-(3-fluorobenzylamino)isonicotinonitrile
[0391] To a scintillation vial containing
2,6-dichloroisonicotinonitrile (500 mg, 2.89 mmol) was added NMP (6
ml) and (3-fluorophenyl)methanamine (868 mg, 6.94 mmol). The
homogenous reaction mixture was capped and heated to 110.degree. C.
in a oil bath for 1 hr. The reaction mixture was diluted with EtOAc
and washed with sat NaHCO.sub.3, H.sub.2O and sat NaCl. The organic
layer was dried Na.sub.2SO.sub.4, filtered and concentrated. The
crude residue was purified by column chromatography on silica gel
(0-20% EtOAc/Hexane) to give
2-chloro-6-(3-fluorobenzylamino)isonicotinonitrile (750 mg, 95%).
LCMS (m/z): 262.0 (MH.sup.+), retention time=1.03 min.
Step 2. Preparation of
5'-chloro-2'-fluoro-6-(3-fluorobenzylamino)-2,4'-bipyridine-4-carbonitril-
e
[0392] To a degassed suspension of 2-chloro-6-(3-fluorobenzylamino)
isonicotinonitrile (150 mg, 0.573 mmol) and
5-chloro-2-fluoropyridin-4-ylboronic acid (151 mg, 0.860 mmol) in
DME (5 ml) was added Na.sub.2CO.sub.3 (1.433 ml, 2M, 2.87 mmol) and
Pd(Ph.sub.3P).sub.4 (66.2 mg, 0.057 mmol). The reaction mixture was
capped and heated to 110.degree. C. in an oil bath for 2 hr. The
reaction mixture was diluted with EtOAc and washed with sat
NaHCO.sub.3, and then sat NaCl. The organic layer was dried over
Na2SO4, filtered and concentrated. The resulting residue was
purified by column chromatography on silica gel (0-20%
EtOAc/Hexane) to give
5'-chloro-2'-fluoro-6-(3-fluorobenzylamino)-2,4'-bipyridine-4-carbonitril-
e (95 mg, 47%). LCMS (m/z): 357.0 (MH.sup.+), retention time=1.09
min
Step 3. Preparation of
2'-(trans-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'--
bipyridine-4-carbonitrile
[0393] To a scintillation vial containing
5'-chloro-2'-fluoro-6-(3-fluorobenzylamino)-2,4'-bipyridine-4-carbonitril-
e (72 mg, 0.202 mmol) was added DMSO (3 ml) and
trans-cyclohexane-1,4-diamine (230 mg, 2.018 mmol). The homogenous
yellow reaction mixture was capped and heated to 105.degree. C. in
a oil bath for 3 hr. The reaction mixture was diluted with EtOAc
and washed with sat NaHCO.sub.3, sat NaCl. The organic layer was
dried Na.sub.2SO.sub.4, filtered and concentrated. The crude solid
was purified by Prep HPLC and the collected fractions were combined
and diluted with EtOAc and neutralized with sat NaHCO.sub.3 and
then sat NaCl. The organic layer was dried over Na.sub.2SO.sub.4,
filtered and concentrated to afford
2'-(trans-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'--
bipyridine-4-carbonitrile (73 mg, 80%). LCMS (m/z): 451.2
(MH.sup.+), retention time=0.70 min. 1H NMR (400 MHz, METHANOL-d4)
d ppm 1.34-1.48 (m, 2H) 1.50-1.64 (m, 2H) 2.06-2.22 (m, 4H)
3.08-3.20 (m, 1H) 3.63-3.74 (m, 1H) 4.61 (s, 2H) 6.81 (s, 1H) 6.87
(s, 1H) 6.91-6.99 (m, 1H) 7.02 (s, 1H) 7.04-7.10 (m, 1H) 7.12-7.18
(m, 1H) 7.25-7.36 (m, 1H) 8.00 (s, 1H).
Examples 16 and 17 (Compounds 16 and 17)
2'-(trans-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'-b-
ipyridine-4-carboxamide &
2'-(trans-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'--
bipyridine-4-carboxylic acid
##STR00061##
[0394] Step 4. Preparation of
2'-(trans-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'--
bipyridine-4-carboxamide
[0395] To a scintillation vial containing
2'-(trans-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'--
bipyridine-4-carbonitrile (11 mg, 0.024 mmol) and K.sub.2CO.sub.3
(33.7 mg, 0.244 mmol) at 0.degree. C. was added DMSO (1 ml) and
H.sub.2O.sub.2 (10.68 .mu.l, 0.122 mmol). The reaction mixture was
capped and stirred at 0.degree. C. for 10 min and rt for 10 min.
The reaction mixture was diluted with EtOAc and washed with
H.sub.2O, sat NaCl. The organic layer was dried over
Na.sub.2SO.sub.4, filtered and concentrated. The crude oil/solid
was purified by reverse phase preparative HPLC to yield a TFA salt
of
2'-(trans-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamin-
o)-2,4'-bipyridine-4-carboxamide (3.5 mg, 25%), LCMS (m/z): 469.2
(MH.sup.+), retention time=0.56 min and
2'-(trans-4-aminocyclohexylamino)-5'-chloro-6-(3-fluorobenzylamino)-2,4'--
bipyridine-4-carboxylic acid (3.2 mg, 22%), LCMS (m/z): 470.2
(MH.sup.+), retention time=0.61 min.
Example 18 (Compound 18)
5'-chloro-N2'-(trans-4-(dimethylamino)cyclohexyl)-N6-(3-fluorobenzyl)-2,4'-
-bipyridine-2',6-diamine
##STR00062##
[0397] Step 1. Preparation of
5'-chloro-N2'-(trans-4-(dimethylamino)cyclohexyl)-N6-(3-fluorobenzyl)-2,4-
'-bipyridine-2',6-diamine: To a scintillation vial containing
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine (7 mg, 0.016 mmol) and formaldehyde (6.12 .mu.l,
0.082 mmol) was added McOH (0.3 ml) and Pd/C (5.25 mg, 4.93
.mu.mol). The reaction mixture was stirred under hydrogen at room
temperature for 16 hours. The reaction mixture was filtered over
celite and concentrated. The crude solid was purified by reverse
phase preparative HPLC to yield
5'-chloro-N2'-(trans-4-(dimethylamino)cyclohexyl)-N6-(3-fluorobenzyl)-2,4-
'-bipyridine-2',6-diamine (2.0 mg, 24%). LCMS (m/z): 454.2
(MH.sup.+), retention time=0.61 min. as a TFA salt after
lypholyzing.
Example 19 (Compound 19)
2-(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)c-
yclohexylamino)ethanol
##STR00063##
[0399] Step 1. Preparation of
N2'-(trans-4-(2-(tert-butyldimethylsilyloxy)ethylamino)cyclohexyl)-5'-chl-
oro-N6-(3-fluorobenzyl)-2,4'-bipyridine-2',6-diamine: To a
scintillation vial containing
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine (17 mg, 0.040 mmol) and K.sub.2CO.sub.3 (22.06 mg,
0.160 mmol) was added DMF (0.3 ml) and
(2-bromoethoxy)(tert-butyl)dimethylsilane (9.55 mg, 0.040 mmol).
The reaction mixture was capped and heated to 75.degree. C. for 7
hr. The reaction mixture was diluted with DCM and washed with
H.sub.2O, sat NaCl. The organic layer was dried over
Na.sub.2SO.sub.4, filtered and concentrated. The crude solid was
purified by reverse phase preparative HPLC. Collected fractions
were combined, neutralized with Saturated NaHCO.sub.3 and extracted
with EtOAc. The organic layer was dried over Na2SO4, filtered,
concentrated and used directly in next step. LCMS (m/z): 584.3
(MH.sup.+), retention time=0.87 min.
[0400] Step 2. Preparation of
2-(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)-
cyclohexylamino)ethanol: To a scintillation vial containing
N2'-(trans-4-(2-(tert-butyldimethylsilyloxy)ethylamino)cyclohexyl)-5'-chl-
oro-N6-(3-fluorobenzyl)-2,4'-bipyridine-2',6-diamine (from step 1)
was added THF (0.300 ml) and TBAF (0.160 ml, 0.319 mmol). The
homogenous reaction mixture was capped, and stirred at ambient
temperature for 3 hours. The reaction mixture was concentrated and
purified by reverse phase preparative HPLC to yield
2-(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)-
cyclohexylamino)ethanol (2.2 mg, 9%). LCMS (m/z): 470.3 (MH.sup.+),
retention time=0.58 min as a TFA salt after lypholyzing. 1H NMR
(400 MHz, METHANOL-d4) .delta. ppm 1.31-1.46 (m, 2H) 1.51-1.67 (m,
1H) 2.21 (d, J=10.56 Hz, 2H) 3.11-3.20 (m, 2H) 3.66-3.77 (m, 1H)
3.77-3.83 (m, 1H) 4.62 (s, 1H) 6.74 (s, 1H) 6.78-6.84 (m, 1H)
6.87-6.92 (m, 1H) 6.96-7.03 (m, 1H) 7.08-7.14 (m, 1H) 7.15-7.21 (m,
1H) 7.31-7.38 (m, 1H) 7.68-7.76 (m, 1H) 8.02 (s, 1H).
Example 20 (Compound 20)
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-(2-(methylsulfonyl)ethylamino)c-
yclohexyl)-2,4'-bipyridine-2',6-diamine
##STR00064##
[0402] Step 1. Preparation of 2-(methylsulfonyl)ethyl
methanesulfonate: To a round-bottom flask containing
2-(methylsulfonyl)ethanol (400 mg, 3.22 mmol) at 0.degree. C. was
added DCM (10 ml) and triethylamine (4.91 .mu.l, 0.035 mmol),
followed by dropwise addition of mesyl chloride (2.96 mg, 0.026
mmol). The ice bath was removed and the reaction mixture was
stirred at ambient temperature for 2 hr. The reaction mixture was
diluted with DCM and washed with sat NaHCO.sub.3 and then sat NaCl.
The organic layer was dried over Na2SO4, filtered and concentrated.
The resulting residue was purified via ISCO (0-60% EtOAc/Hexane) to
yield 2-(methylsulfonyl)ethyl methanesulfonate (400 mg, 61%). LCMS
(m/z): 203.0 (MH.sup.+), retention time=0.37 min.
[0403] Step 2. Preparation of
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-(2-(methylsulfonyl)ethylamino)-
cyclohexyl)-2,4'-bipyridine-2',6-diamine: To a scintillation vial
containing
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine (10 mg, 0.023 mmol) and K2CO3 (8 mg, 0.058 mmol)
was added DMSO (0.5 ml) and 2-(methylsulfonyl)ethyl
methanesulfonate (30 mg). The reaction mixture was capped and
heated to 120.degree. C. in an oil bath for 4 hr. The resulting
solution was purified by reverse phase preparative HPLC to yield
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-(2-(methylsulfonyl)ethylamino)-
cyclohexyl)-2,4'-bipyridine-2',6-diamine (3.7 mg, 24%). LCMS (m/z):
532.2 (MH.sup.+), retention time=0.62 min as a TFA salt after
lypholyzing.
Example 21 (Compound 21)
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-(methylamino)cyclohexyl)-2,4'-b-
ipyridine-2',6-diamine
##STR00065##
[0404] Step 1. Preparation of
(1s,4s)-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)c-
yclohexyl methanesulfonate
[0405] To a round-bottom flask containing
(1s,4s)-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)c-
yclohexanol (obtained following example 2) (85 mg, 0.199 mmol) at
0.degree. C. was added DCM (2 ml) and triethylamine (0.042 ml,
0.299 mmol), followed by dropwise addition of Mesyl Chloride (0.020
ml, 0.259 mmol). The ice bath was removed and the reaction mixture
was stirred at ambient temperature for 2 hr. The reaction mixture
was diluted with DCM and washed with sat NaHCO.sub.3, and then sat
NaCl. The organic layer was dried over Na.sub.2SO.sub.4, filtered
and concentrated to yield
(1s,4s)-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)c-
yclohexyl methanesulfonate (90 mg, 90% yield), LCMS (m/z): 505.3
(MH.sup.+), retention time=0.77 min. The resulting residue was used
in next step without further purification.
Step 2. Preparation of
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-(methylamino)cyclohexyl)-2,4'--
bipyridine-2',6-diamine
[0406] To a scintillation vial containing
(1s,4s)-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)c-
yclohexyl methanesulfonate (20 mg, 0.040 mmol) was added MeOH (1
ml) and methyl amine (0.594 ml, 2M, 1.188 mmol). The reaction
mixture was capped and heated to 70.degree. C. in a oil bath for 16
hr. Solvent was evaporated and recharge the vial with 0.6 ml 30%
methyl amine in ethanol. After heating at 70.degree. C. for another
6 hr, the reaction mixture was concentrated and purified by reverse
phase preparative HPLC to yield
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-(methylamino)cyclohexyl)-2,4'--
bipyridine-2',6-diamine (6.5 mg, 0.015 mmol, 37.3%), LCMS (m/z):
440.3 (MH.sup.+), retention time=0.61 min and
5'-chloro-N2'-(cyclohex-3-enyl)-N6-(3-fluorobenzyl)-2,4'-bipyridine-2',6--
diamine (3.5 mg, 22%) LCMS (m/z): 409.2 (MH.sup.+), retention
time=0.82 min. 1H NMR (400 MHz, METHANOL-d4) .delta. ppm 1.32-1.46
(m, 2H) 1.47-1.62 (m, 2H) 2.20 (d, J=11.35 Hz, 4H) 3.01-3.11 (m,
1H) 3.67-3.78 (m, 1H) 4.64 (s, 2H) 6.81 (s, 1H) 6.88-6.97 (m, 3H)
6.97-7.05 (m, 1H) 7.08-7.14 (m, 1H) 7.16-7.22 (m, 1H) 7.31-7.41 (m,
1H) 7.75-7.83 (m, 1H) 8.05 (s, 1H).
Example 22 (Compound 22)
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-((methylamino)methyl)cyclohexyl-
)-2,4'-bipyridine-2',6-diamine
##STR00066##
[0407] Step 1. Preparation of
(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)cy-
clohexyl)methyl methanesulfonate
[0408] To a round-bottom flask containing
(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)cy-
clohexyl) methanol (obtained following example 2) (102 mg, 0.231
mmol) at 0.degree. C. was added DCM (2 ml) and triethylamine (0.048
ml, 0.347 mmol), followed by dropwise addition of Mesyl Chloride
(0.023 ml, 0.301 mmol). The ice bath was removed and the reaction
mixture was stirred at rt for 2 hr. The reaction mixture was
diluted with DCM and washed with sat NaHCO.sub.3 and then sat NaCl.
The organic layer was dried over Na.sub.2SO.sub.4, filtered and
concentrated to yield
(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)cy-
clohexyl)methyl methanesulfonate (110 mg, 92% yield), LCMS (m/z):
519.2 (MH.sup.+), retention time=0.80 min. The resulting residue
was used in next step without further purification.
Step 2. Preparation of
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-((methylamino)methyl)cyclohexy-
l)-2,4'-bipyridine-2',6-diamine
[0409] To a scintillation vial containing
(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)cy-
clohexyl) methyl methanesulfonate (15 mg, 0.029 mmol) was added
MeOH (1 ml) and a solution of methyl amine (0.144 ml, 0.289 mmol)
in MeOH. The reaction mixture was capped and heated to 70.degree.
C. in a oil bath for 16 hr. The resulting solution was concentrated
and purified by reverse phase preparative HPLC to yield
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-((methylamino)methyl)cyclohexy-
l)-2,4'-bipyridine-2',6-diamine (8.6 mg, 52%), LCMS (m/z): 454.2
(MH.sup.+), retention time=0.64 min as a TFA salt after
lypholyzing. 1H NMR (400 MHz, METHANOL-d4) .delta. ppm 1.15-1.29
(m, 2H) 1.29-1.42 (m, 2H) 1.67-1.80 (m, 1H) 1.86-1.96 (m, 2H)
2.09-2.21 (m, 2H) 2.71 (s, 3H) 2.90 (d, J=7.04 Hz, 2H) 3.58-3.70
(m, 1H) 4.63 (s, 2H) 6.88 (s, 2H) 6.94 (d, J=7.43 Hz, 1H) 6.96-7.03
(m 1H) 7.07-7.13 (m, 1H) 7.15-7.21 (m, 1H) 7.29-7.39 (m, 1H)
7.69-7.78 (m, 1H) 8.01 (s, 1H).
Example 23 (Compound 23)
5'-chloro-N6-(3,5-difluorobenzyl)-N2'-(trans-4-(pyrrolidin-1-yl)cyclohexyl-
)-2,4'-bipyridine-2',6-diamine
##STR00067##
[0410] Step 1. Preparation of
5'-chloro-N6-(3,5-difluorobenzyl)-N2'-(trans-4-(pyrrolidin-1-yl)cyclohexy-
l)-2,4'-bipyridine-2',6-diamine
[0411] To a scintillation vial containing
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3,5-difluorobenzyl)-2,4'-bipy-
ridine-2',6-diamine (12.3 mg, 0.028 mmol) (obtained following
example 2) and K.sub.2CO.sub.3 (15.32 mg, 0.111 mmol) was added
DMSO (0.5 ml) and 1,4-dibromobutane (5.98 mg, 0.028 mmol). The
reaction mixture was capped and heated at 60.degree. C. for 7 hr.
The reaction mixture was diluted with DCM and washed with H.sub.2O,
sat NaCl. The organic layer was dried over Na.sub.2SO.sub.4,
filtered and concentrated. The crude solid was purified by reverse
phase preparative HPLC to yield
5'-chloro-N6-(3,5-difluorobenzyl)-N2'-(trans-4-(pyrrolidin-1-yl)cyclohexy-
l)-2,4'-bipyridine-2',6-diamine (7.8 mg, 46.0%), LCMS (m/z): 498.3
(MH.sup.+), retention time=0.65 min as a TFA salt after
lypholyzing. 1H NMR (400 MHz, METHANOL-d4) .delta. ppm 1.26-1.40
(m, 2H) 1.48-1.62 (m, 2H) 1.85-1.98 (m, 2H) 1.99-2.24 (m, 7H)
2.99-3.14 (m, 4H) 3.51-3.68 (m, 3H) 4.54 (s, 2H) 6.69-6.80 (m, 3H)
6.81-6.90 (m, 3H) 7.60-7.69 (m, 1H) 7.94 (s, 1H).
Example 24 (Compounds 256+257)
N2'-trans-4-aminocyclohehexyl)-5'-chloro-N6-(((R)-2,2-dimethyltetrahydro-2-
H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine and
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((S)-2,2-dimethyltetrahydro-2-
H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00068##
[0412] Step 1: Preparation of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((R/S)-2,2-dimethyltetrahydro-
-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
[0413] A mixture of
(R/S)-5'-chloro-N-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)-2'-fluor-
o-2,4'-bipyridin-6-amine (35 mg, 0.100 mmol),
trans-cyclohexane-1,4-diamine (91 mg, 0.800 mmol), DIPEA (20.25 mg,
0.200 mmol) in DMSO (0.35 mL) was heated at 109.degree. C. for 16
hr. The mixture was diluted with DMSO, filtered through a syringe
filter and purified by HPLC to give
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((R/S)-2,2-dimethyltetrahydro-
-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine as its
trifluoroacetic acid salt. Yield: 29 mg. LCMS (m/z): 444.2 [M+H]+;
Retention time=0.51 min.
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((R)-2,2-dimethyltetrahydro-2H-
-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine and
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((S)-2,2-dimethyltetrahydro-2-
H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
[0414]
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((R/S)-2,2-dimethyltetr-
ahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
trifluoroacetic acid salt was dissolved in MeOH (2 mL) and filtered
through VariPure.TM. IPE [500 mg per 6 mL tube; 0.9 mmol (nominal);
part no.: PL3540-C603VP], eluted with MeOH (6 mL) and concentrated
in vacuo providing
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((R/S)-2,2-dimethyltetrahydro-
-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine as a colorless
oil. Yield: 20 mg. The enantiomers were resolved by chiral HPLC
[Chiralpak AD column 21.times.50 mm, 20 mic; 20 mg/2 mL EtOH;
heptane/IPA; 85:15 (v:v); 20 mL/min, 330 psi]. Fraction 1: White
solid. Yield: 7.2 mg. Retention time: 10.4 min. [Chiralpak AD-H,
column 4.6.times.100 mm, 5 mic; 20 mg/2 mL EtOH; heptane/IPA; 85:15
(v:v); 1 mL/min]. .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta.
[ppm] 1.07-1.18 (m, 2H) 1.20 (s, 3H) 1.21 (s, 3H) 1.23-1.41 (m, 4H)
1.65-1.74 (m, 2H) 1.90-1.99 (m, 2H) 2.09 (m, 3H) 2.71 (br. s., 1H)
3.19 (d, J=6.65 Hz, 2H) 3.57-3.67 (m, 1H) 3.67-3.74 (m, 2H) 6.52
(d, 1H) 6.61 (s, 1H) 6.71 (d, 1H) 7.42-7.50 (m, 1H) 7.94 (s,
1H).
[0415] Fraction 2: White solid. Yield: 6.6 mg. Retention time: 17.4
min. [Chiralpak AD-H, column 4.6.times.100 mm, 5 mic; 20 mg/2 mL
EtOH; heptane/IPA; 85:15 (v:v); 1 mL/min]. .sup.1H NMR (400 MHz,
METHANOL-d.sub.4) .delta. [ppm] 1.06-1.18 (m, 2H) 1.20 (s, 3H) 1.21
(s, 3H) 1.24-1.42 (m, 4H) 1.63-1.74 (m, 2H) 1.91-2.01 (m, 2H)
2.04-2.19 (m, 3H) 2.75 (br. s., 1H) 3.19 (d, J=7.04 Hz, 2H)
3.57-3.66 (m, 1H) 3.66-3.74 (m, 2H) 6.52 (d, 1H) 6.61 (s, 1H) 6.72
(d, 1H) 7.43-7.50 (m, 1H) 7.94 (s, 1H). Absolute stereochemistry of
compounds in Fraction 1 and Fraction 2 is not determined.
Example 25 (Compound 269)
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-((tetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00069##
[0417] A mixture of
5'-chloro-2',5-difluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine (30 mg, 0.088 mmol) and trans-cyclohexane-1,4-diamine
(81 mg, 0.706 mmol) in DMSO (0.3 mL) under argon in a scaled tube
was heated at 103.degree. C. for 18 hr. The mixture was allowed to
cool to ambient temperature. The mixture was diluted with DMSO and
filtered through a syringe filter. Purification by HPLC provided
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-((tetrahydro-2H-pyran-
-4-yl)methyl)-2,4'-bipyridine-2',6-diamine as its trifluoroacetic
acid salt. Yield: 22.3 mg. LCMS (m/z): 434.1 [M+H]+; Retention
time=0.57 min.
[0418] .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. [ppm] 1.22
(dd, J=12.91, 4.30 Hz, 2H) 1.31-1.65 (m, 6H) 1.87 (ddd, J=11.05,
7.34, 3.91 Hz, 1H) 2.07 (dd, 4H) 3.00-3.13 (m, 1H) 3.24-3.34 (m,
4H) 3.50-3.64 (m, 1H) 3.84 (dd. J=11.15, 2.93 Hz, 2H) 6.79 (dd, 1H)
6.93 (s, 1H) 7.20 (dd, 1H) 7.93 (s, 1H).
Example 26 (Compound 155)
Ethyl
2-(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-a-
mino)cyclohexylamino)oxazole-4-carboxylate
##STR00070##
[0420] A mixture of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine (25 mg, 0.059 mmol), ethyl
2-chlorooxazole-4-carboxylate (12.88 mg, 0.073 mmol), triethylamine
(0.041 mL, 0.293 mmol) in dioxane (1 mL) was heated at 80.degree.
C. for .about.20 hr. The mixture was concentrated in vacuo. The
resulting residue was dissolved in DMSO and purified by HPLC
providing ethyl
2-(trans-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)-
cyclohexylamino)oxazole-4-carboxylate as its trifluoroacetic acid
salt. Yield: 7.1 mg. LCMS (m/z): 565.2 [M+H]+; Retention time=0.85
min.
Example 27 (Compound 156)
5'-chloro-N2'-(trans-4-(6-chloropyrimidin-4-yl-amino)cyclohexyl)-N6-(3-flu-
orobenzyl)-2,4'-bipyridine-2',6-diamine
##STR00071##
[0422] A mixture of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridi-
ne-2',6-diamine (25 mg, 0.059 mmol), 4,6-dichloropyrimidine (10.93
mg, 0.073 mmol), triethylamine (0.020 mL, 0.147 mmol) in dioxane (1
mL) was heated at 80.degree. C. for .about.16 hr. The mixture was
concentrated in vacuo. The resulting residue was dissolved in DMSO
and purified by HPLC providing
5'-chloro-N2'-(trans-4-(6-chloropyrimidin-4-yl-amino)cyclohexyl-
)-N6-(3-fluorobenzyl)-2,4'-bipyridine-2',6-diamine as its
trifluoroacetic acid salt. Yield: 18 mg. LCMS (m/z): 538.1 [M+H]+;
Retention time=0.82 min.
Example 28 (Compound 266)
N2'-(trans-4-aminocyclohexyl)-3,5,5'-trichloro-N6-((tetrahydro-2H-pyran-4--
yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00072##
[0423] Step 1: Preparation of
6-bromo-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
[0424] To a solution of 2-bromo-6-fluoropyridine (3 g, 17.05 mmol)
in DMSO (8 mL) was added (tetrahydro-2H-pyran-4-yl)methanamine
(3.10 g, 20.46 mmol) and triethylamine (5.68 mL, 40.9 mmol). The
mixture was heated at 110.degree. C. for 18 hr. The mixture was
allowed to cool to ambient temperature and diluted with EtOAc. The
organic layer was washed with saturated aqueous NaHCO.sub.3
solution (1.times.), water (1.times.), brine (1.times.), dried over
Na.sub.2SO.sub.4, filtered off and concentrated in vacuo. The
resulting residue was purified by column chromatography over silica
gel providing
6-bromo-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine as a
white solid. Yield: 4.24 g. LCMS (m/z): 270.9/273.0 [M+H]+;
Retention time=0.78 min.
Step 2: Preparation of
6-bromo-3,5-dichloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
[0425] Step 2a: To a solution of
6-bromo-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine (20 g,
74 mmol) in acetonitrile (240 mL) was added NCS (9.85 g, 74 mmol).
The mixture was heated to 80.degree. C. for 3 hr. The reaction
mixture was allowed to cool to ambient temperature and concentrated
in vacuo. The resulting residue was diluted with brine (200 mL) and
extracted with EtOAc (3.times.200 mL). The combined organic layers
were concentrated in vacuo. The resulting residue was purified by
column chromatography [SiO.sub.2, EtOAc/heptane=0/100 to 50/50]
providing
6-bromo-5-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(12 g) and a mixture of
6-bromo-3,5-dichloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine/-
6-bromo-3-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(5 g, ratio.about.2:3).
[0426] Step 2b: To a solution of a mixture of
6-bromo-3,5-dichloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine/-
6-bromo-3-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(4.5 g, ratio.about.2:3) in acetonitrile (40 mL) was added NCS
(1.250 g, 9.36 mmol). The mixture was heated to 80.degree. C. for
50 min. The mixture was allowed to cool to ambient temperature and
concentrated in vacuo. The resulting residue was purified by column
chromatography [SiO.sub.2, 120 g, EtOAc/heptane] providing
6-bromo-3,5-dichloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
as white solid. Yield: 2.25 g. LCMS (m/z): 340.9 [M+H]+; Retention
time=1.11 min.
Step 3: Preparation of
3,5,5'-trichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipy-
ridin-6-amine
[0427] A mixture of
6-bromo-3,5-dichloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(1 g, 2.94 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid (0.774
g, 4.41 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (0.240 g,
0.294 mmol) in DME (12 mL) and 2M aqueous Na.sub.2CO.sub.3 solution
(4 mL, 2.94 mmol) in a sealed tube was heated at 90.degree. C. for
2 hr. The mixture was allowed to cool to ambient temperature and
was diluted with EtOAc (.about.100 mL) and saturated aqueous
NaHCO.sub.3. The separated organic layer was washed with saturated
aqueous NaHCO.sub.3 (2.times.), brine, dried over Na.sub.2SO.sub.4,
filtered off and concentrated in vacuo. The resulting residue was
purified by column chromatography [SiO.sub.2, 80 g,
EtOAc/heptane=0/100 to 30/70 over 25 min] providing
3,5,5'-trichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipy-
ridin-6-amine as a colorless liquid. Yield: 510 mg. LCMS (m/z):
391.9 [M+H]+; Retention time=1.14 min.
Step 4: Preparation of
N2'-(trans-4-aminocyclohexyl)-3,5,5'-trichloro-N6-((tetrahydro-2H-pyran-4-
-yl)methyl)-2,4'-bipyridine-2',6-diamine
[0428] A mixture of
3,5,5'-trichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipy-
ridin-6-amine (35 mg, 0.090 mmol) and trans-cyclohexane-1,4-diamine
(10.23 mg, 0.090 mmol) in DMSO (0.3 mL) under argon in a sealed
tube was heated at 100.degree. C. for 18 hr. The mixture was
allowed to cool to ambient temperature. The mixture was diluted
with DMSO, filtered through a syringe filter. Purification by HPLC
provided
N2'-(trans-4-aminocyclohexyl)-3,5,5'-trichloro-N6-((tetrahydro-2H-pyran-4-
-yl)methyl)-2,4'-bipyridine-2',6-diamine as its trifluoroacetic
acid salt. Yield: 38 mg. LCMS (m/z): 486.0 [M+H]+; Retention
time=0.70 min.
[0429] .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. [ppm] 1.28
(dd, J=13.11, 4.11 Hz, 2H) 1.34-1.48 (m, 2H) 1.49-1.69 (m, 4H)
1.85-2.01 (m, 1H) 2.10 (d, J=12.13 Hz, 2H) 2.15-2.26 (m, 2H)
3.07-3.20 (m, 1H) 3.31-3.40 (m, 4H) 3.65-3.75 (m, 1H) 3.91 (dd,
J=11.35, 2.74 Hz, 2H) 6.59 (s, 1H) 7.69 (s, 1H) 8.02 (s, 1H).
Example 29 (Compound 311)
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((4-methyltetrahydro-2H-pyran-4-
-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00073##
[0430] Step 1: Preparation of
4-methyltetrahydro-2H-pyran-4-carbonitrile (following reference:
WO2005/058860)
[0431] To a solution of tetrahydro-2H-pyran-4-carbonitrile (2 g,
18.00 mmol) in THF (10 mL) at 0-5.degree. C. was slowly added LHMDS
(21.59 mL, 21.59 mmol). The mixture was stirred for 1.5 hr
0.degree. C. Iodomethane (3.37 mL, 54.0 mmol) was added slowly and
stirring was continued for 30 min at .about.0.degree. C. and
.about.2 hr at ambient temperature. The mixture was cooled to
0.degree. C. and carefully diluted with 1N aqueous HCl (30 mL) and
EtOAc (5 mL) and concentrated. The resulting residue was taken up
in diethylether and the separated organic layer was washed with
brine, dried over Na.sub.2SO.sub.4, filtered off and concentrated
in vacuo providing crude 4-methyltetrahydro-2H-pyran-4-carbonitrile
as an orange oil, which was directly used in the next reaction
without further purification. Yield: 1.8 g. LCMS (m/z): 126.1
[M+H]+; Retention time=0.44 min.
Step 2: Preparation of
(4-methyltetrahydro-2H-pyran-4-yl)methanamine
[0432] To a solution of 4-methyltetrahydro-2H-pyran-4-carbonitrile
(1.8 g, 14.38 mmol) in THF (30 mL) was added carefully 1M LAH/THF
(21.57 mL, 21.57 mmol) at 0.degree. C. The reaction mixture was
stirred for 15 min at 0.degree. C., allowed to warm to ambient
temperature and stirred for .about.3 hours at ambient temperature.
To the reaction mixture was carefully added water (0.9 mL), 1N
aqueous NaOH (2.7 mL) and water (0.9 mL) [Caution: gas
development!]. The mixture was vigorously stirred for 30 min. The
precipitate was filtered off and rinsed with THF. The solution was
concentrated in vacuo providing crude
(4-methyltetrahydro-2H-pyran-4-yl)methanamine as a yellowish solid,
which was directly used in the next step without further
purification. Yield: 1.54 g. LCMS (m/z): 130.1 [M+H]+; Retention
time=0.21 min.
Step 3: Preparation of
6-bromo-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
[0433] To a solution of 2-bromo-6-fluoropyridine (619 mg, 3.52
mmol) in DMSO (3 mL) was added
(4-methyltetrahydro-2H-pyran-4-yl)methanamine (500 mg, 3.87 mmol)
and triethylamine (498 mg, 4.93 mmol). The mixture was heated at
110.degree. C. for 18 hr. The mixture was allowed to cool to
ambient temperature and diluted with EtOAc. The organic layer was
washed with saturated aqueous NaHCO.sub.3 solution (1.times.),
water (1.times.), brine (1.times.), dried over Na.sub.2SO.sub.4,
filtered off and concentrated in vacuo. The resulting residue was
purified by column chromatography [SiO.sub.2, 24 g,
EtOAc/heptane=0/100 2 min, 0/100 to 40/60 2-25 min] providing
6-bromo-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
as a white solid. Yield: 750 mg. LCMS (m/z): 285.0/287.0 [M+H]+;
Retention time=0.88 min.
Step 4: Preparation of
5'-chloro-2'-fluoro-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)-2,4'-bip-
yridin-6-amine
[0434] A mixture of
6-bromo-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(750 mg, 2.63 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid (830
mg, 4.73 mmol), PdCl.sub.2(dppf)-CH.sub.2Cl.sub.2Adduct (215 mg,
0.263 mmol) in DME (12 mL) and 2M aqueous Na.sub.2CO.sub.3 (4 mL,
8.00 mmol) in a sealed tube was heated at 103.degree. C. for 4 hr.
The mixture was allowed to cool to ambient temperature and was
diluted with EtOAc (.about.50 mL) and saturated aqueous NaHCO.sub.3
solution. The separated organic layer was washed with saturated
aqueous NaHCO.sub.3 solution (2.times.), dried over
Na.sub.2SO.sub.4, filtered off and concentrated in vacuo. The
resulting residue was purified by column chromatography [SiO.sub.2,
40 g, 20 min, EtOAc/heptane=0/100 for 2 min, then
EtOAc/heptane=5/95 to 50/50 over 18 min, then EtOAc/heptane=50/50]
providing
5'-chloro-2'-fluoro-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)-2,4'-bip-
yridin-6-amine as a colorless oil. Yield: 691 mg. LCMS (m/z): 336.2
[M+H]+; Retention time=0.66 min.
Step 5: Preparation of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((4-methyltetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine
[0435] A mixture of
5'-chloro-2'-fluoro-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)-2,4'-bip-
yridin-6-amine (50 mg, 0.149 mmol), trans-cyclohexane-1,4-diamine
(136 mg, 1.191 mmol), DIPEA (30.1 mg, 0.298 mmol) in DMSO (0.5 mL)
was heated at 107.degree. C. for 16 hr. The mixture was diluted
with EtOAc and saturated aqueous NaHCO.sub.3 solution. The
separated aqueous layer was extracted with EtOAc (2.times.). The
combined organic layers were dried over Na.sub.2SO.sub.4, filtered
off and concentrated in vacuo. The resulting residue was dissolved
in DMSO/water (1/1), filtered through a syringe filter and purified
by HPLC providing
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((4-methyltetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine as its trifluoroacetic
acid salt. Yield: 59.5 mg. LCMS (m/z): 430.3 [M+H]+; Retention
time=0.48 min.
[0436] .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. [ppm] 1.13
(s, 3H) 1.33-1.49 (m, 4H) 1.49-1.68 (m, 4H) 2.06-2.23 (m, 4H)
3.07-3.22 (m, 1H) 3.37 (s, 2H) 3.60-3.69 (m, 2H) 3.70-3.80 (m, 3H)
6.77 (s, 1H) 6.90 (d, J=7.04 Hz, 1H) 7.12 (d, J=9.00 Hz, 1H)
7.81-7.91 (m, 1H) 8.09 (s, 1H).
Example 30 (Compound 312)
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-((4-methyltetrahydro-2-
H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00074##
[0437] Step 1: Preparation of
3,6-difluoro-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
[0438] A mixture of 2,3,6-trifluoropyridine (858 mg, 6.45 mmol),
(4-methyltetrahydro-2H-pyran-4-yl)methanamine (1000 mg, 7.74 mmol)
and triethylamine (2.158 mL, 15.48 mmol) in NMP (16 mL) was heated
at 70.degree. C. for 1 hr. The reaction mixture was allowed to
ambient temperature and was diluted with EtOAc (.about.100 mL),
brine (.about.50 mL) and water (.about.50 mL). The separated
organic layer was washed with brine (1.times.), 0.3N aqueous HCl
(2.times.), saturated aqueous NaHCO.sub.3 solution (1.times.),
brine (1.times.), dried over Na.sub.2SO.sub.4, filtered off and
concentrated in vacuo to provide crude
3,6-difluoro-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
as a colorless oil, which was directly used in the next reaction
without further purification. Yield: 1.4 g. LCMS (m/z): 243.1
[M+H]+; Retention time=0.86 min.
Step 2: Preparation of
3-fluoro-6-methoxy-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)pyridin-2--
amine
[0439] To a solution of
3,6-difluoro-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(1.4 g, 5.78 mmol) in MeOH (14 mL) was added sodium methoxide (25
wt. % in MeOH, 7 mL, 30.8 mmol). The mixture was heated in a steel
bomb at 135.degree. C. for 3 days. The mixture was cooled to
ambient temperature and concentrated in vacuo. The resulting
residue was taken up in water (200 mL), and the resulting
precipitate was filtered off and rinsed with water. The solid was
dissolved in DCM. The organic solution was washed with brine, dried
over Na.sub.2SO.sub.4, filtered off and concentrated in vacuo. The
resulting residue was purified by column chromatography [SiO.sub.2,
80 g, 20 min, EtOAc/heptane=0/100 for 2 min, then
EtOAc/heptane=5/95 to 25/75 over 23 min, EtOAc/heptane=25/75]
providing
3-fluoro-6-methoxy-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)pyridin-2--
amine as an off-white solid. Yield: 1.22 g. LCMS (m/z): 255.1
[M+H]+; Retention time=0.89 min.
Step 3: Preparation of
5-fluoro-6-((4-methyltetrahydro-2H-pyran-4-yl)methyl)aminopyridin-2-ol
[0440] To
3-fluoro-6-methoxy-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)p-
yridin-2-amine in acetonitrile (12 mL) was added sodium iodide
(4.24 g, 28.3 mmol) and slowly TMS-Cl (3.62 mL, 28.3 mmol). The
mixture was heated to reflux (oil bath: 83.degree. C.) for 4 hr.
The mixture was allowed to cool to ambient temperature and was
diluted with EtOAc and saturated aqueous NaHCO.sub.3 solution. The
mixture was vigorously stirred for 15 min and acidified with 0.5N
aqueous NaHSO.sub.4 solution and stirring was continued for 5 min.
The mixture was neutralized with saturated aqueous NaHCO.sub.3
solution. The separated aqueous phase was extracted with EtOAc
(3.times.). The combined organic layers were dried over sodium
sulfate, filtered off and concentrated in vacuo. The resulting
residue was purified by column chromatography [SiO.sub.2, 40 g, 25
min, EtOAc/heptane=5/95 for 2 min, 5/95 to 50/50 over 18 min, then
50/50] providing
5-fluoro-6-((4-methyltetrahydro-2H-pyran-4-yl)methyl)aminopyrid-
in-2-ol as colorless highly viscous oil. Yield: 420 mg. LCMS (m/z):
241.1 [M+H]+; Retention time=0.55 min.
Step 4: Preparation of
5-fluoro-6-((4-methyltetrahydro-2H-pyran-4-yl)methyl)aminopyridin-2-yl
trifluoromethanesulfonate
[0441] To a solution of
5-fluoro-6-((4-methyltetrahydro-2H-pyran-4-yl)methyl)aminopyridin-2-ol
(420 mg, 1.748 mmol) and triethylamine (0.731 mL, 5.24 mmol) in DCM
(16 mL) was added trifluoromethanesulfonic anhydride (0.443 mL,
2.62 mmol) slowly at 0.degree. C. The mixture was stirred for 2 hr
at 0.degree. C. and poured carefully into ice-cooled saturated
aqueous NaHCO.sub.3 solution. The separated aqueous layer was
extracted with DCM (2.times.). The combined organic layers were
dried over Na.sub.2SO.sub.4, filtered off and concentrated in
vacuo. The resulting residue was purified by column chromatography
[SiO.sub.2, 24 g, EtOAc/heptane=5/95 for 2 min, then
EtOAc/heptane=5/95 to 40/60 over 13 min, then EtOAc/heptane=40/60]
providing
5-fluoro-6-((4-methyltetrahydro-2H-pyran-4-yl)methyl)aminopyrid-
in-2-yl trifluoromethanesulfonate as colorless oil. Yield: 600
mg.
Step 5: Preparation of
5'-chloro-2',5-difluoro-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)-2,4'-
-bipyridin-6-amine
[0442] A mixture of
5-fluoro-6-((4-methyltetrahydro-2H-pyran-4-yl)methyl)aminopyridin-2-yl
trifluoromethanesulfonate (600 mg, 1.611 mmol),
5-chloro-2-fluoropyridin-4-ylboronic acid (565 mg, 3.22 mmol),
PdCl.sub.2(dppf)-CH.sub.2Cl.sub.2 adduct (132 mg, 0.161 mmol) in
DME (8 mL) and 2M aqueous Na.sub.2CO.sub.3 (3 mL, 6.00 mmol) in a
sealed tube was heated at 102.degree. C. for 10 hr. The mixture was
cooled to ambient temperature and was diluted with EtOAc
(.about.100 mL) and saturated aqueous NaHCO.sub.3 solution. The
separated organic layer was washed with saturated aqueous
NaHCO.sub.3 solution (2.times.), dried over Na.sub.2SO.sub.4,
filtered off and concentrated in vacuo. The resulting residue was
purified by column chromatography [SiO.sub.2, 40 g,
EtOAc/heptane=0/100 for 3 min, EtOAc/heptane=0/100 to 30/70 over 17
min, then EtOAc/heptane=30/70] providing
5'-chloro-2',5-difluoro-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)-2,4'-
-bipyridin-6-amine as a colorless oil. Yield: 490 mg. LCMS (m/z):
354.2 [M+H]+; Retention time=1.05 min.
Step 6: Preparation of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-((4-methyltetrahydro--
2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
[0443] A mixture of
5'-chloro-2',5-difluoro-N-((4-methyltetrahydro-2H-pyran-4-yl)methyl)-2,4'-
-bipyridin-6-amine (50 mg, 0.141 mmol),
trans-cyclohexane-1,4-diamine (129 mg, 1.131 mmol), DIPEA (28.6 mg,
0.283 mmol) in DMSO (0.5 mL) was heated at 107.degree. C. for 16
hr. The mixture was diluted with EtOAc and saturated aqueous
NaHCO.sub.3 solution. The separated aqueous layer was extracted
with EtOAc (2.times.). The combined organic layers were dried over
Na.sub.2SO.sub.4, filtered off and concentrated in vacuo. The
resulting residue was dissolved in DMSO/water (1/1), filtered
through a syringe filter and purified by HPLC providing
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-((4-methyltetrahydro--
2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine as its
trifluoroacetic acid salt. Yield: 61.3 mg. LCMS (m/z): 448.2
[M+H]+; Retention time=0.62 min.
[0444] .sup.1H NMR (400 MHz, METHANOL-d.sub.4) .delta. [ppm] 1.06
(s, 3H) 1.28-1.54 (m, 4H) 1.54-1.65 (m, 4H) 2.06-2.25 (m, 4H)
3.09-3.22 (m, 1H) 3.49 (s, 2H) 3.57-3.72 (m, 3H) 3.72-3.81 (m, 2H)
6.86 (dd, 1H) 6.92 (s, 1H) 7.31 (dd, 1H) 7.99 (s, 1H)
Example 31 (Compound 313)
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((4-fluorotetrahydro-2H-pyran-4-
-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00075##
[0445] Step 1: Preparation of
4-fluorotetrahydro-2H-pyran-4-carbaldehyde (as described in
WO2009/011836)
[0446] Step 1a: To a solution of DIPEA (6.12 mL, 35.0 mmol) in DCM
(80 mL) was added trimethylsilyl trifluoromethanesulfonate (7.79 g,
35.0 mmol) followed by a solution of
tetrahydro-2H-pyran-4-carbaldehyde (2 g, 17.52 mmol) in DCM (80 mL)
at 0.degree. C. Upon completion of the addition, the reaction
mixture was allowed to stir at ambient temperature for 2 hr. The
mixture was concentrated in vacuo and the resulting residue was
treated with hexane (200 mL). The precipitate was filtered off and
the solution was concentrated in vacuo providing crude
trimethylsilyl ether, which was directly used in the next step
without further purification.
[0447] Step 1b: To a solution of crude trimethylsilyl ether in DCM
(100 mL) was added dropwise a solution of
N-fluorobenzenesulfonimide (5.53 g, 17.52 mmol), dissolved in DCM
(50 mL), at 0.degree. C. The mixture was stirred for 3 hr at
ambient temperature and the crude solution of
4-fluorotetrahydro-2H-pyran-4-carbaldehyde was directly used in the
next reaction.
Step 2: Preparation of
6-bromo-N-((4-fluorotetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
[0448] To 6-bromopyridin-2-amine (3.03 g, 17.50 mmol) was added the
crude solution of 4-fluorotetrahydro-2H-pyran-4-carbaldehyde in
DCM. To the mixture was added acetic acid (1.002 mL, 17.50 mmol)
and sodium triacetoxyborohydride (5.56 g, 26.3 mmol) in portions.
The mixture was stirred for 2 hr at ambient temperature. The
mixture was diluted carefully with saturated aqueous NaHCO.sub.3
solution. The separated aqueous layer was extracted with DCM
(1.times.). The combined organic layers were washed with water
(1.times.), saturated aqueous NaHCO.sub.3 solution (1.times.) and
concentrated in vacuo. The solid resulting residue was dissolved in
DCM (100 mL) and 3M aqueous HCl (60 mL). The separated organic
layer was extracted with 3M aqueous HCl (3.times.20 mL). The
combined acidic layers were washed with DCM (1.times.). Solid
NaHCO.sub.3 was added carefully to the acidic solution [Caution:
gas development!] until pH>.about.8. The aqueous mixture was
extracted with DCM (2.times.) and EtOAc (2.times.). The combined
organic layers were concentrated in vacuo. The resulting residue
was dissolved in EtOAc. The solution was washed with 0.3M aqueous
HCl, and brine, dried over Na.sub.2SO.sub.4, filtered off and
concentrated in vacuo. The resulting residue was purified by column
chromatography [SiO.sub.2, 40 g, EtOAc/heptane=5/95 for 3 min, then
EtOAc/heptane=5/95 to 30/70 over 15 min, then EtOAc/heptane=30/70]
providing
6-bromo-N-((4-fluorotetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
as a white solid. Yield: 1.82 g. LCMS (m/z): 288.9/291.0 [M+H]+;
Retention time=0.84 min.
Step 3: Preparation of
5'-chloro-2'-fluoro-N-((4-fluorotetrahydro-2H-pyran-4-yl)methyl)-2,4'-bip-
yridin-6-amine
[0449] A mixture of
6-bromo-N-((4-fluorotetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(1 g, 3.46 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid (1.092
g, 6.23 mmol), PdCl.sub.2(dppf)-CH.sub.2Cl.sub.2 adduct (0.282 g,
0.346 mmol) in DME (13 mL) and 2M aqueous Na.sub.2CO.sub.3 (5.19
mL, 10.38 mmol) in a sealed tube was heated at 100.degree. C. for 2
hr. The mixture was cooled to ambient temperature and was diluted
with EtOAc (.about.50 mL) and saturated aqueous NaHCO.sub.3. The
separated organic layer was washed with saturated aqueous
NaHCO.sub.3 (2.times.), dried over Na.sub.2SO.sub.4, filtered off
and concentrated in vacuo. The resulting residue was purified by
column chromatography [SiO.sub.2, 80 g. EtOAc/heptane=5/95 for 4
min, then EtOAc/heptane=5/95 to 50/50 over 18 min, then
EtOAc/heptane=50/50] providing
5'-chloro-2'-fluoro-N-((4-fluorotetrahydro-2H-pyran-4-yl)methyl)-2,4'-bip-
yridin-6-amine as a colorless oil. Yield: 1.00 g. LCMS (m/z): 340.1
[M+H]+; Retention time=0.67 min.
Step 4: Preparation of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((4-fluorotetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine
[0450] A mixture of
5'-chloro-2'-fluoro-N-((4-fluorotetrahydro-2H-pyran-4-yl)methyl)-2,4'-bip-
yridin-6-amine (75 mg, 0.221 mmol) and
trans-cyclohexane-1,4-diamine (202 mg, 1.766 mmol) in DMSO (1 mL)
under argon in a sealed tube was heated at 103.degree. C. for 18
hr. The mixture was cooled to ambient temperature and diluted with
EtOAc and water. The separated organic layer was washed with
saturated aqueous NaHCO.sub.3 solution and concentrated in vacuo.
The resulting residue was dissolved in DMSO/water (.about.2/1),
filtered through a syringe filter. Purification by HPLC provided
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((4-fluorotetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine as its trifluoroacetic
acid salt. The material was dissolved in MeOH (.about.3 mL),
filtered through VariPure.TM. IPE [500 mg per 6 mL tube; 0.9 mmol
(nominal); part no.: PL3540-C603VP], eluted with MeOH (15 mL) and
concentrated in vacuo. The resulting residue was dissolved in
acetonitrile/water (.about.3/1) and lyophilized providing
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((4-fluorotetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine. Yield: 58 mg. LCMS
(m/z): 434.2 [M+H]+; Retention time=0.50 min.
[0451] .sup.1H NMR (400 MHz. METHANOL-d.sub.4) .delta. [ppm] 1.32
(d, J=9.78 Hz, 4H) 1.73-1.88 (m, 4H) 1.91-1.99 (m, 2H) 2.08 (d,
J=9.78 Hz, 2H) 2.67-2.78 (m, 1H) 3.57-3.73 (m, 5H) 3.75-3.84 (m,
2H) 6.60 (d, J=8.61 Hz, 1H) 6.63 (s, 1H) 6.78 (d, J=7.43 Hz, 1H)
7.34-7.55 (m, 1H) 7.94 (s, 1H).
Example 32 (Compound 152)
N2'-((1S,3S,4S)-4-amino-3-methylcyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)--
2,4'-bipyridine-2',6-diamine/N2'-((1R,3R,4R)-4-amino-3-methylcyclohexyl)-5-
'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridine-2',6-diamine
##STR00076##
[0452] Step 1: Preparation of 4-(dibenzylamino)cyclohexanol
[0453] To a mixture of 4-aminocyclohexanol (3.51 g, 23.15 mmol) and
K.sub.2CO: (12.80 g, 93 mmol) in acetonitrile (100 mL) was added
benzylbromide (5.64 mL, 47.5 mmol) and the mixture was stirred at
reflux for 17 hr. The crude mixture was concentrated in vacuo and
the resulting residue was dissolved in water and EtOAc. The
separated aqueous layer was extracted with EtOAc
(2.times..about.100 mL). The combined organic layers were washed
with brine, dried over Na.sub.2SO.sub.4, filtered off and
concentrated in vacuo providing crude 4-(dibenzylamino)cyclohexanol
as a viscous oil, which was directly used in the next step without
further purification. Yield: 6.12 g. LCMS (m/z): 296.1 [M+H]+;
Retention time=0.59 min.
Step 2: Preparation of 4-(dibenzylamino)cyclohexanone (following
reference WO96/07657)
[0454] To a solution of oxalylic acid (2.03 mL, 20.31 mmol) in DCM
(80 mL) at -60.degree. C. was added dropwise DMSO (3.46 mL, 48.8
mmol). After stirring for 5 min, a solution of
4-(dibenzylamino)cyclohexanol (6 g, 20.31 mmol) in DCM (40 mL) was
added slowly. The mixture was stirred for 15 min and NEt.sub.3
(14.3 mL, 103 mmol) was added slowly. After stirring for 15 min the
ice bath was removed and the mixture was stirred for additional 16
hr. The mixture was diluted with water (100 mL). The separated
organic layer was washed with brine (1.times..about.75 mL), dried
over Na.sub.2SO.sub.4, filtered off and concentrated in vacuo. The
resulting residue was purified by column chromatography [SiO.sub.2,
120 g, EtOAc/hexane=10/90 to 50/50] providing
4-(dibenzylamino)cyclohexanone as a white solid. Yield: 5.5 g. LCMS
(m/z): 294.1 [M+H]+; Retention time=0.58 min.
Step 3: Preparation of
(2S,4S)-4-(dibenzylamino)-2-methylcyclohexanone/(2R,4R)-4-(dibenzylamino)-
-2-methylcyclohexanone
[0455] A solution of 4-(dibenzylamino)cyclohexanone (4 g, 13.63
mmol) in THF (27 mL) was added to KHMDS/toluene (32.7 mL, 16.36
mmol) at ambient temperature. The mixture was stirred for 15 min at
ambient temperature. Triethylborane (1M in THF, 17.72 mL, 17.72
mmol) was added dropwise and the mixture was allowed to stir an
additional 30 min. Iodomethane (1.6 mL, 25.7 mmol) was added and
the mixture was stirred for 20 hr at ambient temperature. Aqueous
1M NaOH solution was added (.about.25 mL) and the mixture was
vigorously stirred for 3 hr. The mixture was extracted with EtOAc
(4.times..about.100 mL) and the combined organic layers were washed
with brine, dried over Na.sub.2SO.sub.4, filtered off, and
concentrated in vacuo. The resulting residue was purified by column
chromatography [SiO.sub.2, 125 g, EtOAc/hexane=0/100 to 20/80].
Fractions were combined and concentrated in vacuo providing
(2S,4S)-4-(dibenzylamino)-2-methylcyclohexanone/(2R,4R)-4-(dibenzylamino)-
-2-methylcylohexanone as a highly viscous oil, which became
partially a white solid. Yield: 3.1 g. LCMS (m/z): 308.2[M+H]+;
Retention time=0.65 min (major isomer). Ratio major/minor isomer:
.about.9:1.
Step 4: Preparation of
(1R,2S,4S)-4-(dibenzylamino)-2-methylcyclohexanol/(1S,2R,4R)-4-(dibenzyla-
mino)-2-methylcyclohexanol
[0456] To a solution of
(2S,4S)-4-(dibenzylamino)-2-methylcyclohexanone/(2R,4R)-4-(dibenzylamino)-
-2-methylcyclohexanone (3.1 g, 10.08 mmol) in THF (55 mL) at
-78.degree. C. was added L-selectride (15.13 mL, 15.13 mmol)
dropwise. After stirring for 5 min at -78.degree. C. the mixture
was allowed to warm up to 0.degree. C. Stirring was continued for
18 hr as the reaction mixture was warmed from 0.degree. C. to
ambient temperature. The mixture was diluted carefully with 1N aq
NaOH (15 mL) and stirred vigorously for 3 hr. The mixture was
extracted with EtOAc (3.times..about.100 mL). The combined organic
layers were washed with brine (.about.100 mL), dried over
Na.sub.2SO.sub.4, filtered off and concentrated in vacuo. The
resulting residue was purified by column chromatography [SiO.sub.2,
120 g, EtOAc/hexane=0/100 to 20/80 over-25 min; EtOAc/hexane=20/80
to 40/60 over 5 min] providing
(1R,2S,4S)-4-(dibenzylamino)-2-methylcyclohexanol/(1S,2R,4R)-4-(dibenzyla-
mino)-2-methylcyclohexanol as a colorless liquid. Yield: 2.83 g.
LCMS (m/z): 310.3 [M+H]+; Retention time=0.66 min.
Step 5: Preparation of
(1S,3S,4S)-4-azido-N,N-dibenzyl-3-methylcyclohexanamine/(1R,3R,4R)-4-azid-
o-N,N-dibenzyl-3-methylcyclohexanamine
[0457] A mixture of DIAD (5.03 mL, 25.9 mmol) and
triphenylphosphine (6.78 g, 25.9 mmol) in THF (35 mL) was allowed
to form a salt. After 30 min a solution of
(1R,2S,4S)-4-(dibenzylamino)-2-methylcyclohexanol/(1S,2R,4R)-4-(dibenzyla-
mino)-2-methylcyclohexanol (2 g, 6.46 mmol) and diphenyl
phosphorazidate (2.507 mL, 11.63 mmol) in THF (25 mL) was added and
the mixture was stirred for 20 hr at 55.degree. C. The mixture was
cooled to ambient temperature and diluted with EtOAc and brine. The
separated organic layer was dried over Na.sub.2SO.sub.4, filtered
off and concentrated in vacuo providing crude
(1S,3S,4S)-4-azido-N,N-dibenzyl-3-methylcyclohexanamine/(1R,3R,4R)-4-azid-
o-N,N-dibenzyl-3-methylcyclohexanamine as orange oil, which was
directly used in the next step without further purification. LCMS
(m/z): 335.1 [M+H]+; Retention time=0.81 min.
Step 6: Preparation of
(1S,3S,4S)--N1,N1-dibenzyl-3-methylcyclohexane-1,4-diamine/(1R,3R,4R)--N1-
,N1-dibenzyl-3-methylcyclohexane-1,4-diamine
[0458] To a solution of
(1S,3S,4S)-4-azido-N,N-dibenzyl-3-methylcyclohexanamine/(1R,3R,4R)-4-azid-
o-N,N-dibenzyl-3-methylcyclohexanamine (2.174 g, 6.5 mmol) in
acetic acid (50 mL) was added slowly Zn-dust (0.638 g, 9.75 mmol).
The mixture was stirred for 30 min at ambient temperature.
Additional Zn-dust was added (150 mg) and stirring was continued
for .about.15 min. The mixture was diluted carefully with 1N
aqueous HCl and diethylether. The separated aqueous layer was
extracted with diethylether (5.times..about.100 mL). The aquous
layer was partially lyophilized and concentrated to dryness in
vacuo. The resulting residue was diluted with 1N aqueous HCl and
concentration to dryness was repeated. Dilution with 1N HCl and
concentration was repeated. The resulting residue was dissolved in
water/acetonitrile and lyophilized to provide crude
(1S,3S,4S)--N1,N1-dibenzyl-3-methylcyclohexane-1,4-diamine/(1R,3R,4R)--N1-
,N1-dibenzyl-3-methylcyclohexane-1,4-diamine as fluffy white solid.
The crude material was directly used in the next step without
further purification. Yield: 2.292 g. LCMS (m/z): 309.3 [M+H]+;
Retention time=0.50 min.
Step 7: Preparation of tert-butyl
(1S,2S,4S)-4-(dibenzylamino)-2-methylcyclohexylcarbamate/tert-butyl
(1R,2R,4R)-4-(dibenzylamino)-2-methylcyclohexylcarbamate
[0459] To
(1S,3S,4S)--N1,N1-dibenzyl-3-methylcyclohexane-1,4-diamine/(1R,3-
R,4R)--N1,N1-dibenzyl-3-methylcyclohexane-1,4-diamine (1.851 g, 6
mmol) in dioxane (200 mL) and saturated aqueous NaHCO.sub.3
solution (100 mL) was added BOC-anhydride (2.438 mL, 10.50 mmol),
dissolved in dioxane (.about.5 mL). The resulting white suspension
was stirred vigorously for 18 hr. The mixture was extracted with
DCM (4.times.300 mL) and EtOAc (1.times.100 mL). The combined
organic layers were concentrated in vacuo. The resulting residue
was dissolved in EtOAc, washed with brine, dried over
Na.sub.2SO.sub.4, filtered of and concentrated in vacuo. The
resulting residue was purified by column chromatography [SiO.sub.2,
120 g, DCM/MeOH=100/0 to 95/5]. Fractions containing product were
combined, concentrated in vacuo providing tert-butyl
(1S,2S,4S)-4-(dibenzylamino)-2-methylcyclohexylcarbamate/tert-butyl
(1R,2R,4R)-4-(dibenzylamino)-2-methylcyclohexylcarbamate. Yield:
778 mg. LCMS (m/z): 409.2 [M+H]+; Retention time=0.84 min.
Step 8: Preparation of tert-butyl
(1S,2S,4S)-4-amino-2-methylcyclohexylcarbamate/tert-butyl
(1R,2R,4R)-4-amino-2-methylcyclohexylcarbamate
[0460] A mixture of tert-butyl
(1S,2S,4S)-4-(dibenzylamino)-2-methylcyclohexylcarbamate/tert-butyl
(1R,2R,4R)-4-(dibenzylamino)-2-methylcyclohexylcarbamate (750 mg,
1.836 mmol) and Pearlman's catalyst (290 mg, 2.73 mmol) in EtOH (35
mL) was hydrogenated in a steel bomb under H.sub.2-atmosphere
(pressure .about.75 psi) for 16 hr. The steel bomb was flushed with
Argon, Celite and methanol were added. The mixture was filtered and
concentrated in vacuo. The white resulting residue was dissolved in
acetonitrile/water (1:1) and lyophilized giving crude tert-butyl
(1S,2S,4S)-4-amino-2-methylcyclohexylcarbamate/tert-butyl
(1R,2R,4R)-4-amino-2-methylcyclohexylcarbamate, which was directly
used in the next step without further purification. Yield: 412 mg.
LCMS (m/z): 173.2/229.3 [M+H]+; Retention time=0.54 min.
Step 9: Preparation of
N2'-((1S,3S,4S)-4-amino-3-methylcyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-
-2,4'-bipyridine-2',6-diamine/N2'-((1R,3R,4R)-4-amino-3-methylcyclohexyl)--
5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridine-2',6-diamine
[0461] Step 9a: A mixture of Intermediate B (preparation of
intermediate B is described in the intermediate session which is in
front of the Examples) (25 mg, 0.075 mmol), tert-butyl
(1S,2S,4S)-4-amino-2-methylcyclohexylcarbamate/tert-butyl
(1R,2R,4R)-4-amino-2-methylcyclohexylcarbamate (25.8 mg, 0.113
mmol), triethylamine (28 .mu.l, 0.201 mmol) in DMSO (0.25 mL) was
heated at 100.degree. C. for 3 days. The mixture was allowed to
cool to ambient temperature and diluted with EtOAc (20 mL) and
saturated aqueous NaHCO.sub.3 solution (10 mL). The separated
aqueous layer was extracted with EtOAc (3.times.). The combined
organic layers were dried over Na.sub.2SO.sub.4, filtered off and
concentrated in vacuo providing crude tert-butyl
(1S,2S,4S)-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amin-
o)-2-methylcyclohexylcarbamate/tert-butyl
(1R,2R,4R)-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amin-
o)-2-methylcyclohexylcarbamate, which was directly used in the next
step without further purification.
[0462] Step 9b: To a solution of crude tert-butyl
(1S,2S,4S)-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amin-
o)-2-methylcyclohexylcarbamate/tert-butyl
(1R,2R,4R)-4-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amin-
o)-2-methylcyclohexylcarbamate was dissolved in MeOH (3 mL) was
added 4M HCl/dioxane (9 mL. 36.0 mmol). The mixture was stirred for
1 hr and concentrated in vacuo. The resulting residue was dissolved
in DMSO, filtered over a syringe filter and purified by HPLC
providing
N2'-((1S,3S,4S)-4-amino-3-methylcyclohexyl)-5'-chloro-N6-(3-fluorobenzyl)-
-2,4'-bipyridine-2',6-diamine/N2'-((1R,3R,4R)-4-amino-3-methylcyclohexyl)--
5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyridine-2',6-diamine as the
trifluoroacetic acid salt. Yield: 28.1 mg. LCMS (m/z): 440.1
[M+H]+; Retention time=0.62 min.
Example 33 (Compound 224)
5-(2-(trans-4-aminocyclohexylamino)-5-chloropyridin-4-yl)-3-((tetrahydro-2-
H-pyran-4-yl)methyl)aminopyrazine-2-carboxamide
##STR00077##
[0463] Step 1. Preparation of
5-(5-chloro-2-fluoropyridin-4-yl)-3-((tetrahydro-2H-pyran-4-yl)methyl)ami-
nopyrazine-2-carboxamide
[0464]
3-chloro-6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran--
4-yl)methyl)pyrazin-2-amine (0.0342 g, 0.096 mmol), CuCN (0.034 g,
0.383 mmol), and dppf (0.085 g, 0.153 mmol) were dissolved in
dioxane (1.5 ml). The solution was then degassed by sparging with
argon for 5 min. It was then treated with Pd.sub.2(dba).sub.3
(0.035 g, 0.038 mmol). The reaction mixture was then heated at
100.degree. C. for 5 hr. The reaction mixture was filtered through
a pad of Celite then it was concentrated in vacuo to give 0.110 g
of
5-(5-chloro-2-fluoropyridin-4-yl)-3-((tetrahydro-2H-pyran-4-yl)methyl)ami-
nopyrazine-2-carboxamide. LCMS (m/z): 366 (MH.sup.+), retention
time=0.89 min.
Step 2. Preparation of
5-(2-(trans-4-aminocyclohexylamino)-5-chloropyridin-4-yl)-3-((tetrahydro--
2H-pyran-4-yl)methyl)aminopyrazine-2-carboxamide
[0465]
5-(5-chloro-2-fluoropyridin-4-yl)-3-((tetrahydro-2H-pyran-4-yl)meth-
yl)aminopyrazine-2-carboxamide (0.035 g, 0.096 mmol) was dissolved
in DMSO (2 ml). This was treated with 1,4-diaminocyclohexane (0.109
g, 0.957 mmol). The reaction mixture was then heated at 100.degree.
C. for 4 hr. The material was purified by preparative reverse-phase
HPLC to give 0.0053 g of
5-(2-(trans-4-aminocyclohexylamino)-5-chloropyridin-4-yl)-3-((tetrahydro--
2H-pyran-4-yl)methyl)aminopyrazine-2-carboxamide as the TFA salt.
LCMS (m/z): 460.1 (MH.sup.+), retention time=0.54 min.
Example 34 (Compound 231)
trans-N1-(5-chloro-4-(5-methyl-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopy-
razin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
##STR00078##
[0466] Step 1. Preparation of
6-(5-chloro-2-fluoropyridin-4-yl)-3-methyl-N-(tetrahydro-2H-pyran-4-yl-me-
thyl)pyrazine-2-amine
[0467]
3-chloro-6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran--
4-yl)methyl)pyrazin-2-amine (0.0275 g, 0.077 mmol), methylboronic
acid (0.014 g, 0.231 mmol), and sodium carbonate (0.100 ml, 0.200
mmol, 2M aq solution) were dissolved in DME (1.0 ml). The solution
was then degassed by sparging with argon for 5 min. It was then
treated with PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (0.013 g,
0.015 mmol). The reaction mixture was then heated in the microwave
at 105.degree. C. for 20 min. More of the above reagents in the
same amounts were added to the reaction mixture and heating in the
microwave was continued at 115.degree. C. for 20 min. The reaction
mixture was allowed to cool to ambient temperature. It was then
filtered through a pad of Celite. The filtrate was concentrated in
vacuo to yield 0.0497 g of a mixture of
6-(5-chloro-2-fluoropyridin-4-yl)-3-methyl-N-((tetrahydro-2H-pyran-4-yl)p-
yrazine-2-amine and
6-(2-fluoro-5-methylpyridin-4-yl)-3-methyl-N-((tetrahydro-2H-pyran-4-yl-m-
ethyl)pyrazine-2-amine.
Step 2. Preparation of
trans-N1-(5-chloro-4-(5-methyl-6-((tetrahydro-2H-pyran-4-yl)methyl)aminop-
yrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
[0468] The mixture of
6-(5-chloro-2-fluoropyridin-4-yl)-3-methyl-N-((tetrahydro-2H-pyran-4-yl)p-
yrazine-2-amine and
6-(2-fluoro-5-methylpyridin-4-yl)-3-methyl-N-((tetrahydro-2H-pyran-4-yl-m-
ethyl)pyrazine-2-amine (0.025 g, 0.074 mmol) and (0.023 g, 0.074
mmol) respectively was dissolved in DMSO (1 ml). This was treated
with 1,4-diaminocyclohexane (0.085 g, 0.742 mmol). The reaction
mixture was then heated at 100.degree. C. for 18 hr. The material
was purified by preparative reverse-phase HPLC to give 0.0047 g of
trans-N1-(5-chloro-4-(5-methyl-6-((tetrahydro-2H-pyran-4-yl)methyl)aminop-
yrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine as the TFA salt.
LCMS (m/z): 431.2 (MH.sup.+), retention time=0.49 min.
Example 35 (Compound 240)
trans-N.sup.1-(5-chloro-4-(5-cyclopropyl-6-((tetrahydro-2H-pyran-4-yl)meth-
yl)aminopyrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
##STR00079##
[0469] Step 1. Preparation of
6-(5-chloro-2-fluoropyridin-4-tl)-3-cyclopropyl-N-((tetrahydro-2H-pyran-4-
-yl)methyl)pyrazin-2-amine
[0470]
3-chloro-6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran--
4-yl)methyl)pyrazin-2-amine (0.0316 g, 0.088 mmol), potassium
cyclopropyltrifluoroborate (0.026 g, 0.177 mmol), and potassium
phosphate (0.113 g, 0.531 mmol) were dissolved in a mixture of
toluene (1 ml) and H.sub.2O (0.170 ml). The solution was then
degassed by sparging with argon for 5 min. At this time it was
treated with PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (0.014 g,
0.018 mmol). The reaction mixture was then heated in the microwave
at 115.degree. C. for 25 min. The reaction mixture was filtered
through a plug of Celite and the filtrate was concentrated in vacuo
to give 0.0445 g of the crude product. The resulting residue was
subjected to silica gel column chromatography. Elution using 20
EtOAc/80 heptane to 70 EtOAc/30 heptane gave 0.0271 g (84%) of
6-(5-chloro-2-fluoropyridin-4-tl)-3-cyclopropyl-N-((tetrahydro-2H-pyran-4-
-yl)methyl)pyrazin-2-amine. LCMS (m/z): 363.1 (MH.sup.+), retention
time=1.06 min.
Step 2. Preparation of
trans-N.sup.1-(5-chloro-4-(5-cyclopropyl-6-((tetrahydro-2H-pyran-4-yl)met-
hyl)aminopyrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
[0471]
6-(5-chloro-2-fluoropyridin-4-tl)-3-cyclopropyl-N-((tetrahydro-2H-p-
yran-4-yl)methyl)pyrazin-2-amine (0.0267 g, 0.074 mmol) was
dissolved in DMSO (1 ml). This was treated with
1,4-diaminocyclohexane (0.084 g, 0.736 mmol). The reaction mixture
was then heated at 100.degree. C. for 4 hr. Additional
1,4-diaminocyclohexane (0.084 g, 0.736 mmol) and triethylamine
(0.0204 ml, 0.028 g, 0.294 mmol) were added. Heating at 100.degree.
C. was continued for 17 hr. The reaction mixture was purified using
prep HPLC. The material was purified by preparative reverse-phase
HPLC to yield 0.0240 g of
trans-N.sup.1-(5-chloro-4-(5-cyclopropyl-6-((tetrahydro-2H-pyran-4-yl)met-
hyl)aminopyrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine as the
TFA salt. LCMS (m/z): 457.2 (MH.sup.+), retention time=0.60
min.
Example 36 (Compound 241)
[0472]
trans-N.sup.1-(5-chloro-4-(5-ethyl-6-((tetrahydro-2H-pyran-4-yl)met-
hyl)aminopyrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
##STR00080##
Step 1. Preparation of
6-(5-chloro-2-fluoropyridin-4-yl)-3-ethyl-N-((tetrahydro-2H-pyran-4-yl)me-
thyl)pyrazine-2-amine
[0473]
3-chloro-6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran--
4-yl)methyl)pyrazin-2-amine (0.0347 g, 0.097 mmol), ethylboronic
acid (0.014 g, 0.194 mmol), and sodium carbonate (0.126 ml g. 0.253
mmol, 2 M aq solution) were dissolved in DME (1 ml). The solution
was then degassed by sparging with argon for 5 min. At this time it
was treated with PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (0.016 g,
0.019 mmol). The reaction mixture was then heated in the microwave
at 115.degree. C. for 25 min. More ethylboronic acid (0.014 g,
0.194 mmol) and PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (0.016 g,
0.019 mmol) were added. The reaction mixture was then heated in the
microwave at 115.degree. C. for 25 min. The reaction mixture was
filtered through a plug of Celite and the filtrate was concentrated
in vacuo to afford 0.0709 g of crude product. The material was
purified using the Isco with a 4 g SiO2 column. The resulting
residue was subjected to silica gel column chromatography. Elution
using 20 EtOAc/80 heptane to 70 EtOAc/30 heptane gave 0.0049 g
(14%) of
6-(5-chloro-2-fluoropyridin-4-yl)-3-ethyl-N-((tetrahydro-2H-pyran-4-yl)me-
thyl)pyrazine-2-amine. LCMS (m/z): 351.1 (MH.sup.+), retention
time=0.97 min.
Step 2. Preparation of
trans-N.sup.1-(5-chloro-4-(5-ethyl-6-((tetrahydro-2H-pyran-4-yl)methyl)am-
inopyrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
[0474]
6-(5-chloro-2-fluoropyridin-4-yl)-3-ethyl-N-((tetrahydro-2H-pyran-4-
-yl)methyl)pyrazine-2-amine (0.0053 g, 0.015 mmol) was dissolved in
DMSO (1 ml). This was treated with 1,4-diaminocyclohexane (0.017 g,
0.151 mmol). The reaction mixture was then heated at 100.degree. C.
for 4 hr. Additional 1,4-diaminocyclohexane (0.017 g, 0.151 mmol)
and triethylamine (0.0084 ml, 0.012 g, 0.060 mmol) were added.
Heating at 100.degree. C. was continued for 17 hr. The reaction
mixture was purified using prep HPLC. The material was purified by
preparative reverse-phase HPLC to give 0.0040 g of
trans-N.sup.1-(5-chloro-4-(5-ethyl-6-((tetrahydro-2H-pyran-4-yl)methyl)am-
inopyrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine as the TFA
salt. LCMS (m/z): 445.2 (MH.sup.+), retention time=0.54 min.
Example 37 (Compound 255)
[0475]
trans-N1-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)amino-3-(-
trifluoromethyl)pyrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
##STR00081##
Step 1. Preparation of
6-chloro-5-iodo-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
[0476] 6-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
(0.250 g, 1.098 mmol) was dissolved in a mixture of DMSO (4.30 ml)
and H.sub.2O (0.105 ml). It was cooled to 0.degree. C. in an ice
bath and was then treated with N-iodosuccinimide (0.247 g, 1.098
mmol) by portion-wise addition. Once the addition was complete the
reaction mixture was stirred at ambient temperature for 24 hr.
Additional NIS (0.025 g, 0.111 mmol) was added. Stirring at ambient
temperature was continued for 24 hr. The reaction mixture was
diluted with H.sub.2O (50 ml). This was extracted with EtOAc
(3.times.50 ml). The organic layers were combined and washed with
brine (1.times.50 ml). The oranic layer was dried
(Na.sub.2SO.sub.4), filtered, and the solvent removed in vacuo to
give 0.410 g of crude product. The resulting residue was subjected
to silica gel column chromatography. Elution using 30 EtOAc/70
heptane to 100 EtOAc gave 0.2144 g (55%) of
6-chloro-5-iodo-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine.
LCMS (m/z): 353.9 (MH.sup.+), retention time=0.92 min. .sup.1H NMR
(400 MHz, CHLOROFORM-d) d ppm 1.37 (qd, 2H) 1.59 (s, 2H) 1.67 (d,
J=12.91 Hz, 2H) 1.77-1.94 (m, J=14.87, 7.63, 7.63, 3.52 Hz, 1H)
3.25 (t, J=6.46 Hz, 2H) 3.39 (td, J=11.74, 1.96 Hz, 2H) 4.00 (dd,
J=11.15, 3.72 Hz, 2H) 4.80 (br. s., 1H) 7.62 (s, 1H).
Step 2. Preparation of
t-butyl-6-chloro-5-iodopyrazin-2-yl((tetrahydro-2H-pyran-4-yl)methyl)carb-
amate
[0477]
6-chloro-5-iodo-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
(0.0801 g, 0.227 mmol) was dissolved in anhydrous DMF and placed
under nitrogen. It was then treated with sodium hydride (0.0109 g.
0.272 mmol, 60% dispersion in mineral oil) followed by
di-t-butyldicarbonate (0.099 g, 0.453 mmol). The reaction mixture
was then stirred at 50.degree. C. for 24 hr. More NaH (0.0109 g,
0.072 mmol) and Boc.sub.2O (0.099 g, 0.453 mmol) were added. The
reaction mixture was then heated at 70.degree. C. for 18 hr. The
reaction mixture was cooled to ambient temperature, and then it was
poured into brine (25 ml). This was extracted with EtOAc
(3.times.25 ml). The combined extracts were washed with H.sub.2O
(3.times.25 ml) followed by brine (1.times.25 ml). The organic
layer was dried (Na.sub.2SO.sub.4), filtered, and the solvent
removed in vacuo to yield 0.0846 g of crude product. The resulting
residue was subjected to silica gel column chromatography. Elution
using 25 EtOAc/75 heptane to 75 EtOAc/25 heptane gave 0.0569 g
(55%) of
t-butyl-6-chloro-5-iodopyrazin-2-yl((tetrahydro-2H-pyran-4-yl)methyl)carb-
amate. LCMS (m/z): 454.0 (MH.sup.+), retention time=1.20 min.
.sup.1H NMR (400 MHz, CHLOROFORM-d) d ppm 1.28-1.46 (m, 4H)
1.46-1.64 (m, 26H) 1.81-2.02 (m, 2H) 3.26-3.42 (m, 3H) 3.86 (d,
J=7.04 Hz, 3H) 3.96 (dd, J=11.54, 2.93 Hz, 3H) 8.86 (s, 1H).
Step 3. Preparation of
6-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)-5-(trifluoromethyl)pyrazin--
2-amine
[0478]
t-butyl-6-chloro-5-iodopyrazin-2-yl((tetrahydro-2H-pyran-4-yl)methy-
l)carbamate (0.0569 g, 0.125 mmol), methyl
2-chloro-2,2-difluoroacetate (0.047 ml, 0.063 g, 0.439 mmol),
potassium fluoride (0.015 g, 0.251 mmol), and copper (I) iodide
(0.100 g, 0.527 mmol) were dissovled in anhydrous DMF (0.80 ml) and
placed under argon. The reaction mixture was then heated at
115.degree. C. for 17 hr. It was allowed to cool to ambient
temperature. The reaction mixture was filtered through a pad of
Celite. The filtrate was poured into brine (25 ml). This was
extracted with EtOAc (3.times.25 ml). The combined extracts were
washed with H.sub.2O (1.times.25 ml) followed by brine (1.times.25
ml). The organic layer was dried (Na.sub.2SO.sub.4), filtered, and
the solvent removed in vacuo to give 0.0401 g of crude product. The
resulting residue was subjected to silica gel column
chromatography. Elution using 25 EtOAc/75 heptane to 100 EtOAc gave
0.0569 g (55%) of
6-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)-5-(trifluoromethyl)pyrazin--
2-amine. LCMS (m/z): 296.0 (MH.sup.+), retention time=0.93 min. 1H
NMR (400 MHz, CHLOROFORM-d) d ppm 1.39 (qd, J=12.33, 4.50 Hz, 2H)
1.68 (d, J=11.35 Hz, 3H) 1.80-2.00 (m, J=14.87, 7.63, 7.63, 3.52
Hz, 1H) 3.32-3.47 (m, 4H) 4.01 (dd, J=11.35, 3.52 Hz, 2H) 5.26 (br.
s., 1H) 7.76 (s, 1H).
Step 4. Preparation of
6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)methyl)-5--
(trifluoromethyl)pyrazin-2-amine
[0479]
6-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)-5-(trifluoromethyl)py-
razin-2-amine (0.020 g, 0.068 mmol),
5-chloro-2-fluoropyridin-4-ylboronic acid (0.036 g, 0.203 mmol),
and sodium carbonate (0.088 ml, 0.176 mmol, 2 M in H.sub.2O) were
dissolved in DME (0.70 ml). The solution was then degassed by
sparging with argon for 5 min. It was then treated with
PdCl.sub.2(dppf) CH.sub.2Cl.sub.2 adduct (0.011 g, 0.014 mmol). The
reaction mixture was then heated in a microwave at 110.degree. C.
for 25 min. Boronic acid (.about.0.036 g, 0.203 mmol) and
PdCl.sub.2(dppf) CH.sub.2Cl.sub.2 adduct (.about.0.011 g, 0.014
mmol) were added. Heating in the microwave was continued at
110.degree. C. for 25 min. The reaction mixture was then filtered
through a pad of Celite. The filtrate was then concentrated in
vacuo to give 0.0759 g of crude product. The resulting residue was
subjected to silica gel column chromatography. Elution using 25
EtOAc/75 heptane to 100 EtOAc gave 0.0178 g (67%) of
6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)methyl)-5--
(trifluoromethyl)pyrazin-2-amine. LCMS (m/z): 391.1 (MH.sup.+),
retention time=0.96 min.
Step 5. Preparation of
trans-N.sup.1-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)amino-3-(t-
rifluoromethyl)pyrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
[0480]
6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)meth-
yl)-5-(trifluoromethyl)pyrazin-2-amine (0.0178 g, 0.046 mmol) was
dissolved in anhydrous DMSO (1.0 ml) and charged to a microwave
vial. This was treated with trans-cyclohexane-1,4-diamine (0.052 g,
0.456 mmol). The reaction mixture was then heated at 100.degree. C.
for 18 hr. The reaction mixture was allowed to cool to ambient
temperature. The material was purified by preparative reverse-phase
HPLC to give 0.0086 g (32%) of
trans-N.sup.1-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)a-
mino-3-(trifluoromethyl)pyrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
as the TFA salt. LCMS (m/z): 485.3 (MH.sup.+), retention time=0.63
min.
Example 38 (Compound 260)
N2'-(trans-4-aminocyclohexyl)-3-chloro-5'-fluoro-N6-((tetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00082##
[0481] Step 1. Preparation of 2,5-difluoropyridin-4-ylboronic
acid
[0482] Diisopropylamine (1.74 ml, 1.24 g, 12.20 mmol) was dissolved
in anhydrous THF (22 ml) and placed under argon. The solution was
cooled to -20.degree. C. and then treated with n-butyllithium (7.66
ml, 12.25 mmol, 1.6 M in hexanes) by slow addition over 10 min. The
newly formed LDA (LDA=lithium diisopropylamide, this acronyl should
be listed in the general session) was then cooled to -78.degree. C.
and treated with a solution of 2,5-difluoropyridine (1.05 ml, 1.33
g, 11.56 mmol) dissolved in anhydrous THF (3 ml) by slow addition
over 30 min. Once the addition was complete the reaction mixture
was allowed to stir at -78.degree. C. for 4 hr. At this time the
reaction mixture was treated with a solution of triisopropyl borate
(5.90 ml, 4.78 g, 25.4 mmol) dissolved in anhydrous THF (8.6 ml) by
dropwise addition. Once the addition was complete the reaction
mixture was allowed to warm to ambient temperature then stirred at
ambient temperature for an additional hour. The reaction mixture
was then quenched by adding 4% aq NaOH (34 ml). The layers were
separated and the aqueous layer was cooled in an ice bath. It was
then acidified to pH=4 with 6N HCl (.about.10 ml) not letting the
temperature go above 10.degree. C. This was then extracted with
EtOAc (3.times.50 ml). The extracts were then washed with brine
(1.times.50 ml), dried (Na.sub.2SO.sub.4), filtered, and the
solvent removed in vacuo. The resulting residue was triturated with
Et2O to give 0.8084 g (44%) of 2,5-difluoropyridin-4-ylboronic
acid.
Step 2. Preparation of
3-chloro-2',5'-difluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine
[0483]
6-bromo-5-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amin-
e (0.500 g, 1.64 mmol), 2,5-difluoropyridin-4-ylboronic acid (0.260
g, 1.64 mmol), and sodium carbonate (2.45 ml, 4.91 mmol, 2 M in
H.sub.2O) were dissolved in DME (7.36 ml). The solution was then
degassed by sparging with argon for 5 min. It was then treated with
PdCl.sub.2(dppf) CH.sub.2Cl.sub.2 adduct (0.267 g, 0.327 mmol). The
reaction mixture was then heated in the microwave at 105.degree. C.
for 25 min. More boronic acid (0.260 g, 1.64 mmol) and
PdCl.sub.2(dppf) CH.sub.2Cl.sub.2 adduct (0.267 g, 0.327 mmol), and
H.sub.2O (.about.2 ml) were added. Heating in the microwave was
continued at 110.degree. C. for 30 min. The reaction mixture was
then filtered through a pad of Celite. The filtrate was then
concentrated in vacuo to give 1.2090 g of crude product. The
resulting residue was subjected to silica gel column
chromatography. Elution using 10 EtOAc/90 heptane to 80 EtOAc/20
heptane gave 0.3584 g (65%) of
3-chloro-2',5'-difluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine. LCMS (m/z): 340.0 (MH.sup.+), retention time=0.90 min.
1H NMR (400 MHz, CHLOROFORM-d) d ppm 1.37 (qd, 3H) 1.60 (br. s.,
2H) 1.68 (d, J=12.91 Hz, 3H) 1.84 (ddd, J=11.15, 7.24, 4.30 Hz, 1H)
3.21 (t, J=6.26 Hz, 2H) 3.32-3.45 (m, 3H) 4.00 (dd, J=11.15, 3.72
Hz, 2H) 4.74 (br. s., 1H) 6.45 (d, J=9.00 Hz, 1H) 6.99-7.07 (m, 1H)
7.51 (d, J=8.61 Hz, 1H) 8.12 (s, 1H).
Step 3. Preparation of
N2'-(trans-4-aminocyclohexyl)-3-chloro-5'-fluoro-N6-((tetrahydro-2H-pyran-
-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
[0484] 3-chloro-2',5'-di
fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridin-6-amine
(0.0509 g, 0.150 mmol) was dissolved in anhydrous DMSO (3.0 ml) and
charged to a microwave vial. This was treated with
trans-cyclohexane-1,4-diamine (0.171 g, 1.498 mmol). The reaction
mixture was then heated at 100.degree. C. for 18 hr. More
trans-cyclohexane-1,4-diamine (0.171 g, 1.498 mmol) was added and
the reaction mixture was stirred at 120.degree. C. for 18 hr. The
reaction mixture was allowed to cool to ambient temperature. The
material was purified by preparative reverse-phase HPLC to give
0.2410 g (30%) of
N2'-(trans-4-aminocyclohexyl)-3-chloro-5'-fluoro-N6-((tetrahydro-2H-pyran-
-4-yl)methyl)-2,4'-bipyridine-2',6-diamine as the TFA salt. LCMS
(m/z): 434.2 (MH.sup.+), retention time=0.55 min.
Example 39 (Compound 282)
N2'-(trans-4-aminocyclohexyl)-5'-chloro-3-fluoro-N6-((tetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00083##
[0486] Step 1. Preparation of
trans-N1-(5'-chloro-3,6-difluoro-2,4'-bipyridin-2'-yl)cyclohexane-1,4-dia-
mine: To a solution of 5'-chloro-2',3,6-trifluoro-2,4'-bipyridine
(95 mg, 0.388 mmol) in DMSO (2.5 mL) was added
trans-1,4-diaminocyclohexane (177 mg, 1.55 mmol). The mixture was
stirred at 90.degree. C. for 2 hr. The cooled reaction mixture was
diluted with ethyl acetate and washed with water. The organic layer
was dried (Na2SO4), filtered, and concentrated to give 137 mg of
crude
trans-N1-(5'-chloro-3,6-difluoro-2,4'-bipyridin-2'-yl)cyclohexane-1,4-dia-
mine which was used without further purification. LCMS (m/z): 339.0
(MH+), retention time=0.54 min
[0487] Step 2. Preparation of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-3-fluoro-N6-((tetrahydro-2H-pyran-
-4-yl)methyl)-2,4'-bipyridine-2',6-diamine: To a solution of
trans-N1-(5'-chloro-3,6-difluoro-2,4'-bipyridin-2'-yl)cyclohexane-1,4-dia-
mine (79 mg, 0.388 mmol) in DMSO (1.5 ml) was added
4-aminomethyltetrahydropyran (161 mg, 1.40 mmol). The mixture was
irradiated by microwave at 180.degree. C. for 1 hr in a sealed
microwave vial. The crude reaction mixture was purified by reverse
phase HPLC and lyophilized to give
N2'-(trans-4-aminocyclohexyl)-5'-chloro-3-fluoro-N6-((tetrahydro-2H-pyran-
-4-yl)methyl)-2,4'-bipyridine-2',6-diamine as its TFA salt. LCMS
(m/z): 434.2 (MH+), retention time=0.57 min.; 1H NMR (400 MHz,
DMSO-d6) .delta. ppm 1.07-1.32 (m, 2H) 1.32-1.49 (m, 1H) 1.59 (d,
J=12.91 Hz, 1H) 1.68-1.83 (m, 1H) 1.96 (dd, 2H) 2.93-3.04 (m, 1H)
3.06 (d, J=6.65 Hz, 1H) 3.24 (t, J=10.76 Hz, 1H) 3.54-3.70 (m, 1H)
3.82 (dd, J=10.96, 2.74 Hz, 1H) 6.53 (s, 1H) 6.57 (dd, J=9.19, 2.93
Hz, 1H) 7.41 (t, 1H) 7.79 (d, J=3.91 Hz, 2H) 8.04 (s, 1H)
Example 40 (Compound 283)
5'-chloro-3-fluoro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tetr-
ahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00084##
[0489] Preparation of
5'-chloro-3-fluoro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tet-
rahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine: To a
mixture of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-3-fluoro-N6-((tetrahydro-2H-py-
ran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine trifluoroacetate (30
mg, 0.055 mmol) and sodium carbonate (23 mg, 0.22 mmol) in DMSO
(0.75 ml) was added p-toluenesulfonic acid 2-methoxyethyl ester (15
mg, 0.066 mmol). The mixture was stirred at 85.degree. C. for 20 hr
in a sealed microwave vial. The cooled reaction mixture was
filtered. The filtrate was purified by reverse phase HPLC and
lyophilized to give 5.0 mg of
5'-chloro-3-fluoro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tet-
rahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine as its
TFA salt. LCMS (m/z): 492.2 (MH+), retention time=0.57 min.; 1H NMR
(400 MHz, CHLOROFORM-d) .delta. ppm 1.10-1.46 (m, 6H) 1.64-1.74 (m,
2H) 1.86 (br. s., 2H) 1.95-2.09 (m, 2H) 2.09-2.26 (m, 2H) 2.58 (br.
s., 1H) 2.88 (t, J=5.09 Hz, 2H) 3.17 (t, J=6.26 Hz, 2H) 3.29-3.45
(m, 5H) 3.53 (t, J=5.09 Hz, 3H) 4.00 (dd, J=11.35, 3.52 Hz, 2H)
4.34-4.47 (m, 1H) 4.54-4.68 (m, 1H) 6.35-6.48 (m, 2H) 7.31 (t,
J=8.80 Hz, 1H) 8.11 (s, 1H).
Example 41 (Compound 286)
N2'-(trans-4-aminocyclohexyl)-3-bromo-5'-chloro-N6-((tetrahydro-2H-pyran-4-
-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00085##
[0491] Preparation of
N2'-(trans-4-aminocyclohexyl)-3-bromo-5'-chloro-N6-((tetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine: To a solution of
3-bromo-5'-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bip-
yridin-6-amine (100 mg, 0.250 mmol) in DMSO (1 mL) was added
trans-1,4-diaminocyclohexane (114 mg, 0.998 mmol). The mixture was
stirred at 110.degree. C. for 19 hr. The crude reaction mixture was
purified by reverse phase HPLC and lyophilized to give 51 mg of
N2'-(trans-4-aminocyclohexyl)-3-bromo-5'-chloro-N6-((tetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine as its TFA salt. LCMS
(m/z): 494.2/496.1 (MH+), retention time=0.61 min; 1H NMR (400 MHz,
DMSO-d6) d ppm 1.06-1.31 (m, 4H) 1.31-1.49 (m, 2H) 1.49-1.64 (m,
2H) 1.64-1.82 (m, 1H) 1.85-2.11 (m, 4H) 2.93-3.12 (m, 3H) 3.22 (t,
J=10.96 Hz, 2H) 3.61 (t, J=10.76 Hz, 1H) 3.81 (dd, J=11.35, 2.74
Hz, 2H) 6.39 (s, 1H) 6.48 (d, 1H) 6.82 (br. s., 1H) 6.94 (br. s.,
1H) 7.59 (d, J=9.00 Hz, 1H) 7.78 (d, J=3=3.91 Hz, 2H) 8.02 (s,
1H)
Example 42 (Compound 288)
(R)-3-(trans-4-(5'-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2,4'-b-
ipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoropropan-2-ol
##STR00086##
[0493] Step 1. Preparation of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)met-
hyl)-2,4'-bipyridine-2',6-diamine: To a solution of
5'-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridin-6-
-amine (500 mg, 1.55 mmol) in DMSO (7 mL) was added
trans-1,4-diaminocyclohexane (710 mg, 6.22 mmol). The mixture was
stirred at 110.degree. C. for 19 hr. The cooled reaction mixture
was diluted with water and extracted with ethyl acetate. The
combined extracts were washed sequentially with water and brine,
dried over sodium sulfate, filtered, and concentrated to give 600
mg of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)met-
hyl)-2,4'-bipyridine-2',6-diamine. LCMS (m/z): 416.1 (MH+),
retention time=0.48 min.
[0494] Step 2. Preparation of
(R)-3-(trans-4-(5'-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2,4'--
bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoropropan-2-ol:
To a solution of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)met-
hyl)-2,4'-bipyridine-2',6-diamine (50 mg, 0.120 mmol) in 2-propanol
(0.8 mL) was added (R)-(+)-3,3,3-trifluoro-1,2-epoxypropane (10.4
uL, 0.120 mmol). The mixture was stirred at 60.degree. C. for 17
hr. The reaction mixture was concentrated. The resulting residue
was purified by reverse phase HPLC and lyophilized to give 63 mg of
(R)-3-(trans-4-(5'-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2,4'--
bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoropropan-2-ol
as its TFA salt. LCMS (m/z): 528.3 (MH+), retention time=0.53 min;
1H NMR (400 MHz, DMSO-d6) .delta. ppm 1.08-1.34 (m, 4H) 1.36-1.56
(m, 2H) 1.61 (d, J=12.52 Hz, 2H) 1.70-1.90 (m, 1H) 2.04 (d, J=9.39
Hz, 3H) 2.13 (d, J=11.74 Hz, 1H) 2.97-3.19 (m, 4H) 3.24 (t, J=10.76
Hz, 3H) 3.64 (d, J=10.96 Hz, 1H) 3.83 (dd, J=10.96, 2.74 Hz, 2H)
4.36-4.50 (m, 2H) 6.54-6.68 (m, 2H) 6.70 (d, J=7.04 Hz, 0H) 6.94
(br. s., 0H) 7.23 (br. s., 0H) 7.53 (br. s., 0H) 8.04 (s, 0H) 8.76
(br. s., 2H)
Example 43 (Compound 289)
(S)-3-(trans-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2,-
4'-bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoropropan-2-ol
##STR00087##
[0496] Step 1. Preparation of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine: To a solution of
3,5'-dichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine (500 mg, 1.40 mmol) in DMSO (8 mL) was added
trans-1,4-diaminocyclohexane (641 mg, 5.61 mmol). The mixture was
stirred at 95.degree. C. for 38 hr. The cooled reaction mixture was
diluted with water and extracted with ethyl acetate. The combined
extracts were washed sequentially with water and brine, dried over
sodium sulfate, filtered, and concentrated. The crude material was
purified by flash chromatography over silica gel
(dichloromethane/methanol gradient) to give 480 mg of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine. LCMS (m/z): 450.2 (MH+),
retention time=0.55 min.
[0497] Step 2. Preparation of
(S)-3-(trans-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2-
,4'-bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoropropan-2-ol:
To a solution of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine (54 mg, 0.120 mmol) in
2-propanol (0.4 mL) was added
(S)-(-)-3,3,3-trifluoro-1,2-epoxypropane (10.4 uL, 0.120 mmol). The
mixture was stirred at 70.degree. C. for 2 hr. The reaction mixture
was concentrated. The resulting residue was purified by reverse
phase HPLC and lyophilized to give 32 mg of
(S)-3-(trans-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2-
,4'-bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoropropan-2-ol
as its TFA salt. LCMS (m/z): 562.3 (MH+), retention time=0.70 min;
1H NMR (400 MHz, DMSO-d6) .delta. ppm 1.01-1.33 (m, 4H) 1.35-1.65
(m, 4H) 1.64-1.84 (m, 1H) 1.93-2.23 (m, 4H) 2.94-3.18 (m, 4H)
3.17-3.35 (m, 3H) 3.53-3.69 (m, 1H) 3.81 (dd, J=11.35, 2.74 Hz, 2H)
4.33-4.48 (m, 1H) 6.38 (s, 1H) 6.55 (d, 1H) 6.82 (br. s., 1H) 6.93
(br. s., 1H) 7.23 (br. s., 1H) 7.48 (d, 1H) 8.02 (s, 1H) 8.72 (br.
s., 2H)
Example 44 (Compound 292)
3-bromo-5'-chloro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tetra-
hydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00088##
[0499] Preparation of
3-bromo-5'-chloro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tetr-
ahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridin-2',6-diamine: To a
mixture of
N2'-(trans-4-aminocyclohexyl)-3-bromo-5'-chloro-N6-((tetrahydro-2H-pyran--
4-yl)methyl)-2,4'-bipyridine-2',6-diamine (30 mg, 0.061 mmol) and
sodium carbonate (19 mg, 0.18 mmol) in DMSO (0.6 ml) was added
p-toluenesulfonic acid 2-methoxyethyl ester (21 mg, 0.091 mmol).
The mixture was stirred at 85.degree. C. for 20 hr in a sealed
microwave vial. The cooled reaction mixture was filtered. The
filtrate was concentrated and the resulting residue was purified by
reverse phase HPLC and lyophilized to give 3.8 mg of
3-bromo-5'-chloro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((t-
etrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine as its
TFA salt. LCMS (m/z): 554.1 (MH+), retention time=0.61 min.
Example 45 (Compound 295)
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-((R)-3,3,-
3-trifluoro-2-methoxypropylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine
##STR00089##
[0501] Preparation of
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-((R)-3,3-
,3-trifluoro-2-methoxypropylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine-
: To a solution of
3,5'-dichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine (36 mg, 0.10 mmol) in DMSO (0.4 mL) was added
trans-N1-((R)-3,3,3-trifluoro-2-methoxypropyl)cyclohexane-1,4-diamine
(48 mg, 0.20 mmol) and 2,6-lutidine (0.023 mL. 0.20 mmol). The
mixture was stirred at 120.degree. C. for 20 hr. The cooled
reaction mixture was purified by reverse phase HPLC and lyophilized
to give 11.4 mg of
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-((R)-3,3-
,3-trifluoro-2-methoxypropylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine
as its TFA salt. LCMS (m/z): 576.2 (MH+), retention time=0.68
min.
Example 46 (Compound 297)
trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)ami-
no-2,4'-bipyridin-2'-yl-amino)cyclohexanol
##STR00090##
[0503] Preparation of
trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)am-
ino-2,4'-bipyridin-2'-yl-amino)cyclohexanol: To a solution of
tert-butyl
3,5'-dichloro-2'-fluoro-2,4'-bipyridin-6-yl((2,2-dimethyltetrahydro-2H-py-
ran-4-yl)methyl)carbamate (30 mg, 0.062 mmol) in DMSO (0.4 mL) was
added trans-4-aminocyclohexanol (36 mg, 0.31 mmol) and DIEA (0.022
mL, 0.12 mmol). The mixture was stirred at 135.degree. C. for 3 hr.
The cooled reaction mixture was diluted with water and extracted
with ethyl acetate. The combined extracts were dried (Na2SO4),
filtered, and concentrated. The resulting residue was re-dissolved
in trifluoroacetic acid (1 mL), stirred for 15 min at ambient
temperature, and then concentrated under reduced pressure. The
crude resulting residue was purified by reverse phase HPLC and
lyophilized to give 23 mg of
trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)methyl)am-
ino-2,4'-bipyridin-2'-yl-amino)cyclohexanol as its TFA salt. LCMS
(m/z): 479.3 (MH+), retention time=0.72 min; 1H NMR (400 MHz,
DMSO-d6) .delta. ppm 0.96 (d, J=12.91 Hz, 2H) 1.08 (s, 6H)
1.15-1.35 (m, 4H) 1.54 (d, =12.91 Hz, 2H) 1.71-2.10 (m, 5H) 3.00
(d, J=6.65 Hz, 2H) 3.31-3.63 (m, 5H) 6.47 (s, 1H) 6.58 (d, 1H) 7.50
(d, J=9.00 Hz, 1H) 8.05 (s, 1H)
Example 47 (Compound 298)
(2S)-3-(trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)me-
thyl)amino-2,4'-bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoroprop-
an-2-ol
##STR00091##
[0505] Step 1. Preparation of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((2,2-dimethyltetrahydro-2-
H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine: To a solution of
tert-butyl
3,5'-dichloro-2'-fluoro-2,4'-bipyridin-6-yl((2,2-dimethyltetrahydro-2H-py-
ran-4-yl)methyl)carbamate (40 mg, 0.083 mmol) in DMSO (0.4 m L) was
added trans-1,4-diaminocyclohexane (47 mg, 0.41 mmol) and DIEA
(0.029 mL, 0.17 mmol). The mixture was stirred at 120.degree. C.
for 2 hr. The cooled reaction mixture was diluted with water and
extracted with ethyl acetate. The combined extracts were washed
sequentially with water and brine, dried over sodium sulfate,
filtered, and concentrated. The resulting residue was re-dissolved
in trifluoroacetic acid (1 mL), stirred for 15 min at ambient
temperature, and then concentrated under reduced pressure. The
resulting residue was taken up in DCM, washed with saturated
aqueous sodium bicarbonate, dried (Na2SO4), filtered, and
concentrated to give 39 mg of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((2,2-dimethyltetrah-
ydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine. LCMS
(m/z): 478.4 (MH+), retention time=0.64 min.
[0506] Step 2. Preparation of
(S)-3-(trans-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2-
,4'-bipyridin-2'-yl-amino)cyclohexylamino)-1,1,1-trifluoropropan-2-ol:
To a solution of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((2,2-dimethyltetrahydro-2-
H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine (19 mg, 0.040
mmol) in 2-propanol (0.3 mL) was added
(S)-(-)-3,3,3-trifluoro-1,2-epoxypropane (3.4 uL, 0.040 mmol). The
mixture was stirred at 70.degree. C. for 2 hr. The reaction mixture
was concentrated. The resulting residue was purified by reverse
phase HPLC and lyophilized to give 9.1 mg of
(2S)-3-(trans-4-(3,5'-dichloro-6-((2,2-dimethyltetrahydro-2H-pyran-4-yl)m-
ethyl)amino-2,4'-bipyridin-2-yl-amino)cyclohexylamino)-1,1,1-trifluoroprop-
an-2-ol as its TFA salt. LCMS (m/z): 590.5 (MH+), retention
time=0.71 min; 1H NMR (400 MHz, DMSO-d6) .delta. ppm 0.81-1.32 (m,
12H) 1.33-1.66 (m, 4H) 1.82-1.99 (m, 1H) 1.99-2.21 (m, 4H)
2.89-3.04 (m, 2H) 3.04-3.19 (m, 2H) 3.27 (d, J=2.35 Hz, 2H) 4.40
(br. s., 1H) 6.38 (s, 1H) 6.55 (d, J=9.00 Hz, 1H) 6.77 (br. s., 1H)
6.91 (br. s., 1H) 7.21 (br. s., 1H) 7.48 (d, J=9.00 Hz, 1H) 8.03
(s, 1H)
Example 48 (Compound 301)
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(2-(trifluoro-
methoxy)ethylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine
##STR00092##
[0508] Preparation of
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(2-(trifluor-
omethoxy)ethylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine: To a
mixture of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)-
methyl)-2,4'-bipyridine-2',6-diamine (42 mg, 0.10 mmol) and
triethylamine (0.028 mL, 0.20 mmol) in chloroform (0.4 ml) was
added 2-(trifluoromethoxy)ethyl trifluoromethanesulfonate (39 mg,
0.15 mmol). The mixture was stirred at ambient temperature for 1
hr. The reaction mixture was concentrated under reduced pressure,
purified by reverse phase HPLC, and lyophilized to give 32 mg of
5'-chloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(2-(trifluor-
omethoxy)ethylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine as its
TFA salt. LCMS (m/z): 528.4 (MH+), retention time=0.53 min.; 1H NMR
(400 MHz, DMSO-d6) d ppm 1.09-1.34 (m, 4H) 1.35-1.54 (m, 2H)
1.55-1.69 (m, 2H) 1.73-1.89 (m, 1H) 1.94-2.17 (m, 4H) 3.04-3.15 (m,
1H) 3.14-3.20 (m, 2H) 3.20-3.30 (m, 2H) 3.30-3.47 (m, 2H) 3.55-3.72
(m, 1H) 3.84 (dd, J=11.15, 2.54 Hz, 2H) 4.35 (t, J=4.70 Hz, 2H)
6.65 (s, 1H) 6.67-6.83 (m, 2H) 7.05 (br. s., 0H) 7.46-7.68 (m, 0H)
8.06 (s, 0H) 8.82 (d, J=3.52 Hz, 2H)
Example 49 (Compound 302)
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(2-(trifl-
uoromethoxy)ethylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine
##STR00093##
[0510] Preparation of
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(2-(trif-
luoromethoxy)ethylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine:
To a mixture of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine (45 mg, 0.10 mmol) and
triethylamine (0.028 mL, 0.20 mmol) in chloroform (0.4 ml) was
added 2-(trifluoromethoxy)ethyl trifluoromethanesulfonate (39 mg,
0.15 mmol). The mixture was stirred at ambient temperature for 1
hr. The reaction mixture was concentrated under reduced pressure,
purified by reverse phase HPLC, and lyophilized to give 29 mg of
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(2-(trif-
luoromethoxy)ethylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine as
its TFA salt. LCMS (m/z): 562.4 (MH+), retention time=0.67 min.; 1H
NMR (400 MHz, DMSO-d6) .delta. ppm 1.07-1.32 (m, 4H) 1.36-1.52 (m,
2H) 1.58 (d, J=12.91 Hz, 2H) 1.65-1.84 (m, 1H) 2.07 (d, J=10.56 Hz,
4H) 2.99-3.17 (m, 3H) 3.23 (t, J=10.76 Hz, 2H) 3.35 (br. s., 2H)
3.64 (br. s., 1H) 3.72-3.89 (m, 2H) 4.34 (t, J=4.89 Hz, 2H)
6.32-6.47 (m, 1H) 6.49-6.65 (m, 1H) 6.67-7.10 (m, 2H) 7.49 (d,
J=9.00 Hz, 1H) 8.03 (s, 1H) 8.75 (d, J=3.91 Hz, 1H)
Example 50 (Compound 284)
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((2R,6S)-2,6-dimethyltetrahydr-
o-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00094##
[0512] The mixture of
5'-chloro-N-(((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)methyl)-2'-flu-
oro-2,4'-bipyridin-6-amine L (30 mg, 0.08 mmol),
trans-1,4-cyclohexanediamine (49 mg, 0.43 mmol) and triethylamine
(26 mg, 0.25 mmol) in 1.5 ml DMSO was heated in a reaction vessel
at 110.degree. C. in an oil bath for 16 h. Formation of desired
product was confirmed by LC/MS. The reaction mixture solution was
diluted with ethyl acetate, washed with water, dried over sodium
sulfate and concentrated. Crude compound was purified by HPLC to
give desired product as TFA salt. LCMS (m/z): 444.2/446.2 (MH+),
retention time=0.54 min.
Example 51 (Compound 285)
5'-chloro-N6-(((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)methyl)-N2'-(t-
rans-4-(2-methoxyethylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine
##STR00095##
[0514] The mixture of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-(((2R,6S)-2,6-dimethyl
tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
Compound 284 (20 mg, 0.045 mmol), p-toluenesulfonic acid
2-methoxyethyl ester (14 mg, 0.06 mmol) and sodium carbonate (9.6
mg, 0.09 mmol) in 1 ml DMSO was heated in a reaction vessel at
105.degree. C. in an oil bath for 3 h. Formation of desired product
was confirmed by LCMS, MH+ 502/504, 0.58 min, with .about.50%
conversion. Mixture was diluted with ethyl acetate, washed with
water, dried over sodium sulfate, and concentrated. Crude product
was purified by HPLC to give desired product as TFA salt. LCMS
(m/z): 502.2/504.2, retention time=0.56 min.
Example 52 (Compound 191)
N2'-((1R,3R)-3-aminocyclopentyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyri-
dine-2',6-diamine
##STR00096##
[0515] Step 1. Preparation of
6-bromo-N-(3-fluorobenzyl)pyridin-2-amine
[0516] To 2,6-dibromopyridine (7.1 g, 30.0 mmol) was added NMP (16
ml), (3-fluorophenyl)methanamine (4.13 g, 33.0 mmol) and Huenig's
Base (5.76 ml, 33.0 mmol) flushed with argon. The crude reaction
mixture was stirred at 115-120.degree. C. for 168 hr, followed by
LCMS. The crude mixture was cooled, 250 ml of ethyl acetate was
added, washed with saturated sodium bicarbonate (2.times.), water
(2.times.), saturated salt solution (1.times.), dried sodium
sulfate, filtered, concentrate. The crude was purified by silica
gel chromatography using 120 g column, eluting from 0%-20% ethyl
acetate with hexane. The desired fractions were concentrated to
constant mass, giving 7.11 grams of the title compound as a free
base used without further purification. LCMS (m/z): 281.1/283.1
(MH+), retention time=1.03 min.
Step 2. Preparation of
5'-chloro-2'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine
[0517] To 6-bromo-N-(3-fluorobenzyl)pyridin-2-amine (2.0 g, 7.11
mmol) was added 5-chloro-2-fluoropyridin-4-ylboronic acid (1.996 g,
11.38 mmol), PdCl2(dppf).CH2Cl2 adduct (0.465 g, 0.569 mmol), DME
(27 ml) and last 2M sodium carbonate (9.25 ml, 18.50 mmol). The
crude reaction was stirred at 100.degree. C. for 3 hr, followed by
LCMS. The crude mixture was cooled, 25 ml of ethyl acetate and 20
ml of methanol was added, filtered and concentrated to provide a
crude product. The crude was purified by silica gel chromatography
using a 120 g column, eluting from 0%-20%/ethyl acetate with
hexane. The desired fractions were concentrated to constant mass,
giving 1.259 grams of titled compound as free base use with out
further purification. LCMS (m/z): 332.2 (MH+), retention time=0.92
min.
Step 3. Preparation of
(1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)c-
yclopentanol
[0518] To
5'-chloro-2'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine (75
mg, 0.226 mmol), was added (1S,3R)-3-aminocyclopentanol (68.6 mg,
0.678 mmol), NMP (0.75 ml) and triethylamine (0.158 ml, 1.130
mmol). The crude reaction mixture was stirred at 100.degree. C. for
18 hr, and the reaction progress followed by LCMS. The crude
reaction mixture was cooled, filtered, and purified by prep LC. The
product fractions were collected, 50 mL of 1 M NaOH and 50 mL of
EtOAc were added. The aqueous layer was removed, the organic layer
was washed with 50 mL of saturated salt solution, dried over sodium
sulfate, and reduced to constant mass. 28 mg of the desired
compound was obtained. LCMS (m/z): 413.1 (MH+), retention time=0.67
min.; 1H NMR (400 MHz, CHLOROFORM-d, 25.degree. C.) .delta. ppm
1.71 (d, J=14.09 Hz, 1H) 1.75-1.91 (m, 2H) 1.97-2.05 (m, 1H)
2.10-2.16 (m, 1H) 2.61 (br. s., 1H) 4.03-4.18 (m, 1H) 4.39 (tt,
J=4.84, 2.59 Hz, 1H) 4.55 (d, J=5.09 Hz, 2H) 5.19 (br. s., 2H) 6.41
(d, J=8.22 Hz, 1H) 6.55 (s, 1H) 6.90-7.02 (m, 2H) 7.05-7.18 (m, 2H)
7.24-7.34 (m, 1H) 7.43-7.55 (m, 1H) 8.07 (s, 1H).
Step 4. Preparation of
N2'-((1R,3R)-3-aminocyclopentyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-bipyr-
idine-2',6-diamine
[0519] To
(1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-y-
l-amino)cyclopentanol (28 mg, 0.068 mmol) was added DCM (1 ml),
diisopropyl ethylamine (0.030 ml, 0.170 mmol) then mesyl chloride
(5.81 Ail, 0.075 mmol), stirred at ambient temperature for 1 hr,
and followed by LCMS. Another 3 uL of mesyl chloride was added and
the reaction mixture was stirred an additional 30 minutes at
ambient temperature. DCM was removed by rotary evaporation, and
crude
(1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)c-
yclopentyl LCMS (m/z): 491.2 (MH+), retention time=0.76 min. was
redissolved in 2 mL DMF. Sodium azide (8.82 mg, 0.136 mmol) and
diisopropyl ethylamine (0.030 ml, 0.170 mmol) were added, and the
reaction mixture was heated at 50.degree. C. for 18 hours, at which
point only
N2'-((1R,3R)-3-azidocyclopentyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'--
bipyridine-2',6-diamine was observed by LCMS (m/z): 438.2 (MH+),
retention time=0.83 min. The resulting reaction mixture was
partitioned between ethyl acetate and water. The aqueous layer was
removed, and the organic layer was washed with water (1.times.)
then saturated salt solution (1.times.), dried over sodium sulfate,
and reduced to constant mass. Crude
N2'-((1R,3R)-3-azidocyclopentyl)-5'-chloro-N6-(3-fluorobenzyl)-2,4'-
-bipyridine-2',6-diamine (20 mg, 0.046 mmol, LCMS (m/z): 438.2
(MH+), retention time=0.83 min.) was dissolved in 1 mL of methanol,
and 10% palladium on charcoal (4.86 mg, 0.046 mmol) was added under
argon. H2 was bubbled through the solution while stirring for 1 hr
at ambient temperature, and the reaction was followed by LCMS. The
crude reaction mixture was filtered over celite washed with
methanol, reduced, redissolved in DMSO, filtered and purified
through prep LC. The resulting product fractions were combined,
then 50 mL of 1 M NaOH and 50 mL of EtOAC were added. The aqueous
layer was removed, the organic layer was washed with saturated salt
solution, dried over sodium sulfate, and reduced to constant mass.
8 mg of the desired compound was obtained. LCMS (m/z): 412.1 (MH+),
retention time=0.58 min.; 1H NMR (300 MHz. CHLOROFORM-d, 25.degree.
C.) .delta. ppm 1.31-1.54 (m, 2H) 1.71-1.86 (m, 4H) 1.98-2.13 (m,
1H) 2.20-2.35 (m, 1H) 3.54 (qd, J=6.35, 6.15 Hz, 1H) 4.14 (sxt,
J=6.56 Hz, 1H) 4.55-4.67 (m, 3H) 5.11 (t, J=5.86 Hz, 1H) 6.40 (d,
J=8.50 Hz, 1H) 6.56 (s, 1H) 6.88-7.02 (m, 1H) 7.12-7.16 (m, 1H)
7.29-7.34 (m, 1H) 7.47-7.52 (m, 1H) 8.09 (s, 1H).
Example 53 (Compound 205)
(1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)-N-
,N-dimethylcyclopentanecarboxamide
##STR00097##
[0521] Step 1. Preparation of
6-bromo-N-(3-fluorobenzyl)pyridin-2-amine: To 2,6-dibromopyridine
(7.1 g, 30.0 mmol) was added NMP (16 ml),
(3-fluorophenyl)methanamine (4.13 g, 33.0 mmol) and Huenig's Base
(5.76 ml, 33.0 mmol) flushed with argon. The crude reaction mixture
was stirred at 115-120.degree. C. for 168 hr, followed by LCMS. The
crude mixture was cooled, 250 ml of ethyl acetate was added, washed
with saturated sodium bicarbonate (2.times.), water (2.times.),
saturated salt solution (1.times.), dried sodium sulfate, filtered,
concentrate. The crude was purified by silica gel chromatography
using 120 g column, eluting from 0%-20% ethyl acetate with hexane.
The desired fractions were concentrated to constant mass, giving
7.11 grams of the titled compound as a free base used without
further purification. LCMS (m/z): 281.1/283.1 (MH+), retention
time=1.03 min.
[0522] Step 2. Preparation of
5'-chloro-2'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine: To
6-bromo-N-(3-fluorobenzyl)pyridin-2-amine (2.0 g, 7.11 mmol) was
added 5-chloro-2-fluoropyridin-4-ylboronic acid (1.996 g, 11.38
mmol), PdCl2(dppf).CH2Cl2 adduct (0.465 g, 0.569 mmol), DME (27 ml)
and last 2M sodium carbonate (9.25 ml, 18.50 mmol). The crude
reaction mixture was stirred at 100.degree. C. for 3 hr, followed
by LCMS. The crude mixture was cooled, 25 ml of ethyl acetate and
20 ml of methanol was added, filtered and concentrated to crude
product. The crude was purified by silica gel chromatography using
a 120 g column, eluting from 0%-20% ethyl acetate with hexane. The
desired fractions were concentrated to constant mass, giving 1.259
grams of title compound as free base use with out further
purification. LCMS (m/z): 332.2 (MH+), retention time=0.92 min.
[0523] Step 3: Preparation of
(1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)c-
yclopentanecarboxylic acid: To
5'-chloro-2'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine (100
mg, 0.301 mmol), was added (1S,3R)-3-aminocyclopentanecarboxylic
acid (117 mg, 0.904 mmol), powdered potassium hydroxide (85 mg,
1.507 mmol) and dioxane (1 ml). The reaction mixture was stirred at
100.degree. C. for 18 hr in a sealed vessel and followed by LCMS.
The crude reaction mixture was partitioned between 30 mL saturated
ammonium chloride and 30 mL ethyl acetate. The organic layer was
removed, dried over sodium sulfate, and reduced. This was
redissolved in 1.5 mL DMSO, filtered, and purified through prep LC.
The product fractions were combined and extracted with 50 mL ethyl
acetate, which was dried over sodium sulfate, and concentrated to
constant mass. 10 mg of the desired compound was obtained. LCMS
(m/z): 441.2 (MH+), retention time=0.68 min. 1H NMR (400 MHz,
CHLOROFORM-d, 25.degree. C.) .delta. ppm 1.59 (m, 2H) 1.83 (m, 2H)
1.99 (m, 1H) 2.72 (m, 1H) 3.40 (br. s., 1H) 3.78 (br. s., 1H) 4.42
(br. s., 1H) 5.48 (br. s., 1H) 6.29 (d, J=8.22 Hz, 1H) 6.50 (s, 1H)
6.80-6.92 (m, 2H) 6.95-7.10 (m, 2H) 7.16-7.25 (m, 1H) 7.38 (t,
J=8.02 Hz, 1H) 7.89 (s, 1H).
[0524] Step 4. Preparation of
(1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)--
N,N-dimethylcyclopentanecarboxamide: To
(1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)c-
yclopentanecarboxylic acid U-31332-EXP080 (10 mg, 0.023 mmol), 2M
dimethyl amine in THF (0.011 ml, 0.023 mmol),
N1-((ethylimino)methylene)-N3,N3-dimethylpropane-1,3-diamine
hydrochloride (8.70 mg, 0.045 mmol),
3H-[1,2,3]triazolo[4,5-b]pyridin-3-ol (4.32 mg, 0.032 mmol) were
added then dimethylformamide (1 ml) and diisopropyl ethylamine
(0.016 ml, 0.091 mmol) were added, and the reaction mixture was
stirred at ambient temperature for 18 hr and the progress followed
by LCMS. The crude reaction mixture was filtered and purified by
preparative LC. The product fractions were combined, 50 mL of 1M
NaOH and 50 mL of ethyl acetate were added. The organic layer was
removed, washed with 50 mL 1M NaOH, 50 mL saturated salt solution,
dried over sodium sulfate, and reduced to constant mass. 3 mg of
the desired compound was obtained. LCMS (m/z): 468.1 (MH+),
retention time=0.72 min., 1H NMR (300 MHz, CHLOROFORM-d) .delta.
ppm 1.77-2.16 (m, 6H) 2.96 (s, 3H) 3.07 (s, 3H) 3.10-3.25 (m, 1H)
4.29 (m, 1H) 4.56 (d, J=5.27 Hz, 2H) 5.12 (br. s., 1H) 5.87 (br.
s., 1H) 6.38 (d, J=8.50 Hz, 1H) 6.58 (s, 1H) 6.91-7.01 (m, 1H)
7.06-7.20 (m, 1H) 7.26-7.37 (m, 2H) 7.44-7.53 (m, 1H) 8.09 (s,
1H).
Example 54 (Compound 235)
5'-chloro-N6-(3-fluorobenzyl)-N2'-((1R,3S)-3-((methylamino)methyl)cyclopen-
tyl)-2,4'-bipyridine-2',6-diamine
##STR00098##
[0525] Step 1. Preparation of
6-bromo-N-(3-fluorobenzyl)pyridin-2-amine
[0526] To 2,6-dibromopyridine (7.1 g, 30.0 mmol) was added NMP (16
ml), (3-fluorophenyl)methanamine (4.13 g, 33.0 mmol) and Huenig's
Base (5.76 ml, 33.0 mmol) flushed with argon. The crude reaction
mixture was stirred at 115-120.degree. C. for 168 hr, followed by
LCMS. The crude mixture was cooled, 250 ml of ethyl acetate was
added, washed with saturated sodium bicarbonate (2.times.), water
(2.times.), saturated salt solution (1.times.), dried sodium
sulfate, filtered, concentrate. The crude was purified by silica
gel chromatography using 120 g column, eluting from 0%-20% ethyl
acetate with hexane. The desired fractions were concentrated to
constant mass, giving 7.11 grams of the titled compound as a free
base used without further purification. LCMS (m/z): 281.1/283.1
(MH+), retention time=1.03 min.
Step 2. Preparation of
5'-chloro-2'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine
[0527] To 6-bromo-N-(3-fluorobenzyl)pyridin-2-amine (2.0 g, 7.11
mmol) was added 5-chloro-2-fluoropyridin-4-ylboronic acid (1.996 g,
11.38 mmol), PdCl2(dppf).CH2Cl2 adduct (0.465 g, 0.569 mmol), DME
(27 ml), and 2M sodium carbonate (9.25 ml, 18.50 mmol). The crude
reaction mixture was stirred at 100.degree. C. for 3 hr, and the
reaction progress followed by LCMS. The crude mixture was cooled,
25 ml of ethyl acetate and 20 ml of methanol were added, filtered
and concentrated to yield a crude product. The crude was purified
by silica gel chromatography using a 120 g ISCO column, eluting
from 0%-20% ethyl acetate with hexane. The desired fractions were
concentrated to constant mass, giving 1.259 grams of title compound
as free base use with out further purification. LCMS (m/z): 332.2
(MH+), retention time=0.92 min.
Step 3. Preparation of (1R,4S)-tert-butyl
3-oxo-2-azabicyclo[2.2.1]heptane-2-carboxylate
[0528] A mixture of (1S,4R)-2-azabicyclo[2.2.1]hept-5-en-3-one (2
g, 18.33 mmol) and 10% Pd/C (0.780 g, 0.733 mmol) in MeOH (100 ml)
was stirred under atmospheric pressure of H2 at ambient temperature
for 2 hr, and the reaction progress was followed by LCMS. Pd/C was
filtered off over Celite and the filter cake was washed with MeOH.
The combined organics were concentrated to afford crude
(1R,4S)-2-azabicyclo[2.2.1]heptan-3-one. LCMS (m/z): 112.1 (MH+),
retention time=0.30 min. The resulting residue was redissolved in
DCM (100 ml), to which di-tert-butyl dicarbonate (8.51 ml, 36.7
mmol) and DMAP (1.231 g, 10.08 mmol) were added and stirred at
ambient temperature for 18 hr and the reaction progress was
followed by LCMS. Solvent was removed, and the crude reaction
mixture was purified through column chromatography, 10-40%
EtOAc:Heptane. The desired fractions were concentrated to constant
mass, yielding (1R,4S)-tert-butyl
3-oxo-2-azabicyclo[2.2.1]heptane-2-carboxylate (2.99 g, 14.15 mmol)
of a white solid. LCMS (m/z): 156.2 (M-tBu), retention time=0.75
min.
Step 4. Preparation of tert-butyl
(1R,3S)-3-(hydroxymethyl)cyclopentylcarbamate
[0529] (1R,4S)-tert-butyl
3-oxo-2-azabicyclo[2.2.1]heptane-2-carboxylate (2.99 g, 14.15 mmol)
was dissolved in MeOH (40 ml) and cooled to 0.degree. C. Sodium
Borohydride (1.071 g, 28.3 mmol) was added and the reaction was
stirred at 0.degree. C. for 1 hr, and the reaction progress was
followed by LCMS. MeOH was removed and the resulting residue was
partitioned between EtOAc (250 mL) and H2O (250 mL). The organic
layer was washed with brine (250 mL), dried over Na2SO4, and
concentrated under reduced pressure. The crude material was
purified by column chromatography, 50-100% EtOAc in heptane to
yield tert-butyl (1R,3S)-3-(hydroxymethyl)cyclopentylcarbamate
(2.92 g, 13.56 mmol) as a white solid. LCMS (m/z): 160.2 (M-tBu),
retention time=0.65 min.
Step 5. Preparation of ((1S,3R)-3-aminocyclopentyl)methanol
[0530] Tert-butyl (1R,3S)-3-(hydroxymethyl)cyclopentylcarbamate
(2.92 g, 13.56 mmol) was dispersed in H.sub.2O (50 ml) and refluxed
at 100.degree. C. for 18 hr, followed by LCMS. Water was removed by
azeotroping with toluene (50 mL.times.3). Collected
((1S,3R)-3-aminocyclopentyl)methanol (1.92 g, 12.50 mmol) as a
clear, viscous oil which was used without further purification.
LCMS (m/z): 116.1 (MH+), retention time=0.67 min.
Step 6. Preparation of
((1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)-
cyclopentyl)methanol
[0531] To
5'-chloro-2'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine (100
mg, 0.301 mmol) was added DMSO (1 ml),
((1S,3R)-3-aminocyclopentyl)methanol (104 mg, 0.903 mmol) and TEA
(0.21 ml, 1.51 mmol). The crude mixture was stirred at 100.degree.
C. for 20 hours, followed by LCMS. The crude reaction mixture was
cooled, was diluted with EtOAc (60 mL), washed H.sub.2O (60
mL.times.2), brine (60 mL), dried over Na.sub.2SO.sub.4, and
reduced. The crude was adsorbed onto silica gel, and purified by
silica gel chromatography, 40-80% EtOAc/Heptane, 12 g ISCO silica
column, resulting in
((1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)-
cyclopentyl)methanol (101 mg, 0.237 mmol). LCMS (m/z): 427.1 (MH+).
retention time=0.69 min.
Step 7. Preparation of
(1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)c-
yclopentanecarbaldehyde
[0532] In a flame-dried argon purged 20 mL conical flask, oxalyl
chloride (0.025 ml, 0.281 mmol) was dissolved in DCM (0.5 ml) and
cooled to -78.degree. C. under argon. DMSO (0.030 ml, 0.422 mmol)
was dissolved in DCM (0.5 ml) and added dropwise to the previous
solution (I don't see issues here). This was stirred for 30 min at
-78.degree. C.
((1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-yl-amino)-
cyclopentyl)methanol (60 mg, 0.141 mmol) was dissolved in DCM (0.5
ml) and added dropwise to the reaction mixture. The resulting
mixture was stirred for 60 min at -78.degree. C. TEA (0.078 ml,
0.562 mmol) was dissolved in DCM (0.5 ml) and added dropwise to the
reaction mixture, after which the reaction mixture was allowed to
stir and warm to ambient temp over 2 hr. The reaction mixture was
diluted with EtOAc, washed with saturated NH4Cl (30 mL.times.3),
H20 (30 mL), brine (30 mL), dried over Na2SO4 and reduced. The
resulting residue was used without further purification. LCMS
(m/z): 425.2 (MH+), retention time=0.72.
Step 8. Preparation of
5'-chloro-N6-(3-fluorobenzyl)-N2'-((1R,3S)-3-((methylamino)methyl)cyclope-
ntyl)-2,4'-bipyridine-2',6-diamine
[0533] To
(1S,3R)-3-(5'-chloro-6-(3-fluorobenzylamino)-2,4'-bipyridin-2'-y-
l-amino)cyclopentanecarbaldehyde (20 mg, 0.047 mmol) was added
methyl amine in THF (0.5 ml, 1.0 mmol) and DCM (0.5 mL). Acetic
acid (2.69 .mu.l, 0.047 mmol), and sodium triacetoxyborohydride
(14.96 mg, 0.071 mmol) were added and stirred for 2 hr at ambient
temperature, and the reaction progress was followed by LCMS.
Solvents were removed, and the crude reaction mixture redissolved
in 1.5 mL of DMSO, followed by purification using preperative HPLC.
Product fractions were combined and lyophilized to afore
5'-chloro-N6-(3-fluorobenzyl)-N2'-((1R,3S)-3-((methylamino)methyl)cyclope-
ntyl)-2,4'-bipyridine-2',6-diamine (2.5 mg, 0.006 mmol) as a TFA
salt. LCMS (m/z): 440.2 (MH+), retention time=0.62 min.
Example 56 (Compound 212)
N-2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((R)-6-oxaspiro[2.5]octan-1-yl-
)-2,4'-bipyridine-2',6-diamine
##STR00099##
[0534] Step 1: Preparation of
(R)-6-bromo-N-(6-oxaspiro[2.5]octan-1-yl)pyridin-2-amine
[0535] To a solution of 2,6-dibromopyridine (200 mg, 0.84 mmol) in
NMP (0.42 mL) was added (R)-6-oxaspiro[2.5]octan-1-amine
hydrochloride (138 mg, 0.84 mmol) and potassium carbonate (350 mg,
2.53 mmol). The mixture was heated at 110.degree. C. for 18 hr. The
mixture was allowed to cool to ambient temperature and diluted with
EtOAc. The organic layer was washed with saturated aqueous sodium
bicarbonate solution, water, and brine and dried over sodium
sulfate, filtered off and concentrated in vacuo. The resulting
residue was purified by column chromatography [SiO.sub.2, 40 g,
EtOAc/heptane=0/100 to 30/70]. Pure fractions were combined and
concentrated in vacuo giving 210 mg of titled compound. LCMS (m/z):
282.9/284.9 [M+H]+, retention time=0.85 min.
Step 2. Preparation of
(R)-5'-chloro-2'-fluoro-N-(6-oxaspiro[2.5]octan-1-yl)-2,4'-bipyridin-6-am-
ine
[0536] A mixture of
(R)-6-bromo-N-(6-oxaspiro[2.5]octan-1-yl)pyridin-2-amine (C, 100
mg, 0.35 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid (136 mg,
0.77 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (23 mg, 0.028
mmol) in DME (1 mL) and 2M Na.sub.2CO.sub.3 (97 mg, 0.92 mmol) in a
sealed tube was heated at 103.degree. C. for 2 hr. The mixture was
allowed to cool to ambient temperature and was diluted with EtOAc
(.about.25 mL) and MeOH (.about.5 mL), filtered off and
concentrated in vacuo. The resulting residue was purified by column
chromatography [SiO.sub.2, 12 g, EtOAc/heptane=10/90 to 50/50].
Fractions were combined and concentrated in vacuo giving 105 mg of
titled compound. LCMS (m/z): 334.0/336.0 [M+H]+, retention
time=0.64 min.
Step 3. Preparation of
N-2'-(trans-4-aminocyclohexyl)-5'-chloro-N6-((R)-6-oxaspiro[2.5]octan-1-y-
l)-2,4'-bipyridine-2',6-diamine
[0537] A mixture of
(R)-5'-chloro-2'-fluoro-N-(6-oxaspiro[2.5]octan-1-yl)-2,4'-bipyridin-6-am-
ine (15 mg, 0.045 mmol), trans-cyclohexane-1,4-diamine (10.3 mg,
0.090 mmol), in DMSO (0.2 mmol) in a sealed tube was heated at
110.degree. C. for 18 hr. The mixture was allowed to cool to
ambient temperature. To the reaction mixture was added 0.5 ml of
DMSO, filtered and purified by prep LC. After lyophilisation, 5.0
mg of the titled compound as a TFA salt was obtained. LCMS (m/z):
428.3/430.3 (MH+), retention time=0.46 min.
Example 57 (Compound 230)
N-(4-Amino-cyclohexyl)-5'-chloro-N-(1,1-dioxo-hexahydro-1-thiopyran-4-yl-m-
ethyl)-[2,4']bipyridinyl-6,2'-diamine
##STR00100##
[0538] Step 1. Preparation of toluene-4-sulfonic acid
1,1-dioxo-hexahydro-1-thiopyran-4-yl-methyl ester
[0539] A solution of
(1,1-Dioxo-hexahydro-1-thiopyran-4-yl)-methanol (500 mg, 3.04 mmol)
in pyridine (10 mL) was added 4-methylbenzene-1-sulfonyl chloride
(871 mg, 4.57 mmol). The mixture was stirred at ambient temperature
for 18 hr. The mixture was diluted with EtOAc. The organic layer
was washed with saturated aqueous sodium bicarbonate solution,
water, and brine and dried over sodium sulfate, filtered off and
concentrated in vacuo. The resulting residue was purified by column
chromatography [SiO.sub.2, 12 g, EtOAc/heptane=0/100 to 30/70].
Pure fractions were combined and concentrated in vacuo giving 736
mg of title compound. LCMS (m/z): 319.1 (MH+), retention time=0.69
min.
Step 2. Preparation of
(6-Bromo-pyridin-2-yl)-(1,1-dioxo-hexahydro-1-thiopyran-4-yl-methyl)-amin-
e
[0540] A mixture of toluene-4-sulfonic acid
1,1-dioxo-hexahydro-1-thiopyran-4-yl-methyl ester (736 mg, 2.31
mmol), 6-bromopyridin-2-amine (400 mg, 2.312 mmol), potassium
carbonate (639 mg, 4.62 mmol), sodium hydride (111 mg, 4.62 mmol)
in a sealed tube was heated at 68.degree. C. for 18 hr. The mixture
was allowed to cool to ambient. The mixture was diluted with EtOAc.
The organic layer was washed with saturated aqueous sodium
bicarbonate solution, water, and brine and dried over sodium
sulfate, filtered off and concentrated in vacuo. The resulting
residue was purified by column chromatography [SiO.sub.2, 12 g,
EtOAc/heptane=0/100 to 30/70]. Pure fractions were combined and
concentrated in vacuo giving 240 mg of titled compound. LCMS (m/z):
318.8/320.9 (MH+), retention time=0.71 min.
Step 3. Preparation of
(5'-Chloro-2'-fluoro-[2,4']bipyridinyl-6-yl)-(1,1-dioxo-hexahydro-1-thiop-
yran-4-yl-methyl)-amine
[0541] A mixture of
(6-bromo-pyridin-2-yl)-(1,1-dioxo-hexahydro-1-thiopyran-4-yl-methyl)-amin-
e (238 mg, 0.746 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid
(261 mg, 1.491 mmol), adduct (48.7 mg, 0.060 mmol) in DME (2 mL)
and 2M Na.sub.2CO.sub.3 (205 mg, 1.938 mmol) in a sealed tube was
heated at 103.degree. C. for 2 hr. The mixture was allowed to cool
to ambient temperature and was diluted with EtOAc (.about.25 mL)
and MeOH (.about.5 mL), filtered off and concentrated in vacuo. The
resulting residue was purified by column chromatography [SiO.sub.2,
12 g, EtOAc/heptane=10/90 to 50/50]. Fractions were combined and
concentrated in vacuo giving 150 mg of the title compound. LCMS
(m/z): 370.0/372.0 (MH+); Retention time=0.56 min.
Step 4. Preparation of
N-(4-amino-cyclohexyl)-5'-chloro-N-(1,1-dioxo-hexahydro-1-thiopyran-4-yl--
methyl)-[2,4']bipyridinyl-6,2'-diamine
[0542] A mixture of
(R)-5'-chloro-2'-fluoro-N-(6-oxaspiro[2.5]octan-1-yl)-2,4'-bipyridin-6-am-
ine (40 mg, 0.108 mmol), and trans-cyclohexane-1,4-diamine (124 mg,
1.082 mmol) in DMSO (0.4 mmol) was heated in a sealed tube at
100.degree. C. for 4 hr. The mixture was allowed to cool to ambient
temperature. To the cooled reaction mixture was added 0.5 ml of
DMSO, filtered and purified by prep LC. After lyophilisation, 10.0
mg of the titled compound as a TFA salt was obtained. LCMS (m/z):
464.1/466.1 (MH+), retention time=0.44 min.
Example 58 (Compound 317)
5'-chloro-N6-(dideutero-(tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((-
S)-tetrahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diami-
ne
##STR00101##
[0543] Step 1. Preparation of
dideutero-(tetrahydro-2H-pyran-4-yl)methanamine
[0544] To a solution of tetrahydro-2H-pyran-4-carbonitrile (800 mg,
7.20 mmol) in THF (20 mL) was added aluminum(III) lithium deuteride
at 0.degree. C. The mixture was stirred at 0.degree. C. for 2 hr.
To the stirred reaction mixture was sequentially added 300 uL of
water, 900 .mu.L of 1 N NaOH and 300 .mu.L of water. The mixture
was filtered through a thin layer of celite to remove the solid.
The filtrate was dried over sodium sulfate, filtered off and
concentrated in vacuo giving 700 mg of titled compound. LCMS (m/z):
118.2 [M+H]+, retention time=0.25 min. The crude product was used
directly for next step.
Step 2. Preparation of
6-bromo-N-(dideutero(tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
[0545] To a solution of 2,6-dibromopyridine (1051 mg, 5.97 mmol) in
DMSO (5 mL) was added
dideutero(tetrahydro-2H-pyran-4-yl)methanamine (700 mg, 5.97 mmol)
and diisopropylethylamine (926 mg, 7.17 mmol). The mixture was
heated at 80.degree. C. for 2 hr. The mixture was allowed to cool
to ambient temperature and diluted with EtOAc. The organic layer
was washed with saturated aqueous sodium bicarbonate solution,
water, and brine and dried over sodium sulfate, filtered off and
concentrated in vacuo. The resulting residue was purified by column
chromatography [SiO.sub.2, 40 g, EtOAc/heptane=0/100 to 30/70].
Pure fractions were combined and concentrated in vacuo giving 780
mg of titled compound. LCMS (m/z): 272.9/274.9 [M+H]+, retention
time=0.77 min.
Step 3. Preparation of
5'-chloro-N-(dideutero(tetrahydro-2H-pyran-4-yl)methyl)-2'-fluoro-2,4'-bi-
pyridin-6-amine
[0546] A mixture of
6-bromo-N-(dideutero(tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(500 mg, 1.83 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid (642
mg, 3.66 mmol), PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (120 mg,
0.146 mmol) in DME (1 mL) and 2M Na.sub.2CO.sub.1 (2.38 ml, 4.76
mmol) was heated in a sealed tube at 80.degree. C. for 48 hr. The
mixture was allowed to cool to ambient temperature and was diluted
with EtOAc (.about.25 mL) and MeOH (.about.5 mL), filtered off and
concentrated in vacuo. The resulting residue was purified by column
chromatography [SiO.sub.2, 12 g, EtOAc/heptane=10/90 to 50/50].
Fractions were combined and concentrated in vacuo giving 180 mg of
titled compound. LCMS (m/z): 324.0/325.8 [M+H]+, retention
time=0.58 min.
Step 4. Preparation of
5'-chloro-N6-(dideutero(tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((-
S)-tetrahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diami-
ne
[0547] A mixture of
5'-chloro-N-(dideutero(tetrahydro-2H-pyran-4-yl)methyl)-2'-fluoro-2,4'-bi-
pyridin-6-amine (30 mg, 0.093 mmol),
trans-N1-(((S)-tetrahydrofuran-2-yl)methyl)cyclohexane-1,4-diamine
(60 mg, 0.30 mmol), in DMSO (0.4 mmol) was heated in a sealed tube
at 110.degree. C. for 68 hr. The mixture was allowed to cool to
ambient temperature. To the reaction mixture was added 0.5 ml of
DMSO, filtered and purified by prep LC. After lyophilisation, 10.0
mg of the titled compound as a TFA salt was obtained. LCMS (m/z):
502.3/504.3 (MH+), retention time=0.49 min.
Example 59 (Compound 324)
5'-chloro-5-fluoro-N2'-(trans-4-(oxetan-2-yl-methylamino)cyclohexyl)-N6-((-
tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00102##
[0549] To a stirred solution of
N.sup.2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N.sup.6-((tetrahydr-
o-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine (90 mg, 0.207
mmol)) in DMSO (1.0 ml) was add potassium carbonate (71.7 mg, 0.518
mmol), followed by folllloowwoxetan-2-yl-methyl
4-methylbenzenesulfonate (151 mg, 0.622 mmol). The mixture was
heated at 83.degree. C. for 2 h. The mixture was allowed to cool to
ambient temperature, then diluted with water and then extracted
with EtOAc (.times.3). The organics were combined then washed with
water (.times.2), saturated brine (.times.2), then dried
(Na.sub.2SO.sub.4), filtered and evaporated under reduced pressure.
The resulting residue was purified by reverse phase prep HPLC and
lyophilized to yield titled compound. LCMS (m/z): 504.4/506.5
(MH.sup.+) retention time=0.60 min as a TFA salt.
Example 60 (Compound 222)
trans-4-(5-chloro-4-(5-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyr-
azin-2-yl)pyridin-2-yl-amino)cyclohexanol
##STR00103##
[0550] Step 1. Preparation of
6-bromo-3-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
[0551] To a scintillation vial containing
3,5-dibromo-2-chloropyrazine (1 g, 3.67 mmol) and TEA (1.024 ml,
7.34 mmol) was added MeCN (5 ml) and
(tetrahydro-2H-pyran-4-yl)methanamine (0.557 g, 3.67 mmol). The
homogenous reaction mixture was capped, and heated to 80.degree. C.
in a oil bath for 4 hr. The reaction mixture was concentrated to
dryness, diluted with EtOAc and sequentially washed with sat
NaHCO.sub.3, and sat NaCl. The organic layer was dried
Na.sub.2SO.sub.4, filtered and concentrated. The crude was purified
by column chromatography on silica gel (20% EtOAc/Hexane) to yield
6-bromo-3-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
(688 mg, 2.244 mmol, 61.1% yield), yield), LCMS (m/z): 308.0
(MH.sup.+), retention time=0.94 min, and
6-bromo-5-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
(55 mg, 0.179 mmol, 4.89% yield), LCMS (m/z): 308.0 (MH.sup.+),
retention time=0.91 min.
Step 2. Preparation of
3-chloro-6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)m-
ethyl)pyrazin-2-amine
[0552] To a degassed suspension of
6-bromo-3-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
(358 mg, 1.168 mmol), Na2CO3 (1.518 ml. 3.04 mmol) and
5-chloro-2-fluoropyridin-4-ylboronic acid (307 mg, 1.752 mmol) in
DME (5 ml) was added PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (76
mg, 0.093 mmol). The reaction mixture was capped in a flask and
heated to 100.degree. C. for 4 hr an oil bath. The reaction mixture
was diluted with EtOAc and washed with H.sub.2O saturated NaCl. The
organic layer was dried Na.sub.2SO.sub.4, filtered and
concentrated. The crude oil/solid was purified column
chromatography on silica gel (30% EtOAc/Hexane) to yield
3-chloro-6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran--
4-yl)methyl)pyrazin-2-amine (160 mg, 0.448 mmol, 38.4% yield), LCMS
(m/z): 357.0 (MH.sup.+), retention time=1.02 min.
Step 3. Preparation of
trans-4-(5-chloro-4-(5-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopy-
razin-2-yl)pyridin-2-yl-amino)cyclohexanol
[0553] To a scintillation vial containing
3-chloro-6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)m-
ethyl)pyrazin-2-amine (20 mg, 0.056 mmol) was added DMSO (1 ml) and
trans-4-aminocyclohexanol (32.2 mg, 0.280 mmol). The reaction
mixture was capped and heated to 120.degree. C. in an oil bath for
3 hr. The reaction product was purified by reverse phase
preparative HPLC to yield
trans-4-(5-chloro-4-(5-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopy-
razin-2-yl)pyridin-2-yl-amino)cyclohexanol (2.2 mg, 4.86 .mu.mol,
8.69% yield), LCMS (m/z): 452.1 (MH.sup.+), retention time=0.76 min
as a TFA salt after lypholyzing. 1H NMR (400 MHz, METHANOL-d4)
.delta. ppm 1.17-1.26 (m, 4H) 1.27-1.39 (m, 2H) 1.58 (dd, J=13.11,
1.76 Hz, 2H) 1.84-2.02 (m, 5H) 3.30 (d, J=7.04 Hz, 4H) 3.43-3.61
(m, 2H) 3.84 (dd, J=11.35, 3.13 Hz, 2H) 6.58 (s, 1H) 7.66 (s, 1H)
7.90 (s, 1H).
Example 61 (Compound 223)
trans-N1-(5-chloro-4-(3-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopy-
razin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
##STR00104##
[0555] Step 1. Preparation of
5-chloro-6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)m-
ethyl)pyrazin-2-amine. To a suspension of
6-bromo-5-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
(20 mg, 0.065 mmol), Na.sub.2CO.sub.3 (17.98 mg, 0.170 mmol) and
5-chloro-2-fluoropyridin-4-ylboronic acid (17.16 mg, 0.098 mmol) in
DM E (1 ml) was added PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct
(4.26 mg, 5.22 .mu.mol). The reaction mixture was capped in a flask
and heated to 100.degree. C. for 4 hr an oil bath. The reaction
mixture was diluted with EtOAc and washed with H.sub.2O sat NaCl.
The organic layer was dried Na.sub.2SO.sub.4, filtered and
concentrated. The crude was purified by column chromatography on
silica gel (50% EtOAc/Hexane) to yield
5-chloro-6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)m-
ethyl)pyrazin-2-amine (10 mg, 0.028 mmol, 42.9% yield). LCMS (m/z):
357.0 (MH.sup.+), retention time=0.95 min.
[0556] Step 2. Preparation of
trans-N1-(5-chloro-4-(3-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminop-
yrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine. To a
scintillation vial containing
5-chloro-6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)m-
ethyl) pyrazin-2-amine (10 mg, 0.028 mmol) and TEA (7.80 .mu.l,
0.056 mmol) was added DMSO (1 ml) and trans-cyclohexane-1,4-diamine
(32.0 mg, 0.280 mmol). The resulting homogenous reaction mixture
was capped and heated to 100.degree. C. in an oil bath for 3 hr.
The reaction product was purified by reverse phase preparative HPLC
to yield
trans-N1-(5-chloro-4-(3-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminop-
yrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine (7.7 mg, 0.014
mmol, 48.6% yield), LCMS (m/z): 451.1 (MH.sup.+), retention
time=0.63 min and a TFA salt after lyapholization.
[0557] 1H NMR (400 MHz, METHANOL-d4) .delta. ppm 1.23-1.36 (m, 3H)
1.36-1.49 (m, 2H) 1.51-1.71 (m, 4H), 1.80-1.94 (m, 1H) 2.06-2.25
(m, 4H) 3.08-3.19 (m, 1H) 3.23 (d, J=6.65 Hz, 2H) 3.33-3.43 (m, 2H)
3.66-3.77 (m, 1H) 3.92 (dd, J=11.35, 3.13 Hz, 2H) 6.69 (s, 1H) 7.76
(s, 1H) 8.05 (s, 1H).
Example 62 (Compound 225)
3-chloro-6-(5-chloro-2-(trans-4-(pyrrolidin-1-yl)cyclohexylamino)pyridin-4-
-yl)-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
##STR00105##
[0559] Step 1. Preparation of
3-chloro-6-(5-chloro-2-(trans-4-(pyrrolidin-1-yl)cyclohexylamino)
pyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine:
To a scintillation vial containing
trans-N1-(5-chloro-4-(5-chloro-6-((tetrahydro-2H-pyran-4-yl)methyl)aminop-
yrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine (12 mg, 0.027
mmol) and K2CO3 (3.67 mg, 0.027 mmol) was added DMF (1 ml) and
1,4-dibromobutane (3.15 .mu.l, 0.027 mmol). The reaction mixture
was capped and heated to 60.degree. C. for 3 hr. The crude solution
was concentrated and purified by reverse phase preparative HPLC to
yield
3-chloro-6-(5-chloro-2-(trans-4-(pyrrolidin-1-yl)cyclohexylamino)pyridin--
4-yl)-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine (3.8 mg,
6.13 .mu.mol, 23.07% yield), LCMS (m/z): 505.2 (MH.sup.+),
retention time=0.64 min, and a TFA salt after lyapholization.
[0560] 1H NMR (400 MHz, METHANOL-d4) .delta. ppm 1.26-1.47 (m, 4H)
1.56-1.73 (m, 4H) 2.01 (m, 3H) 2.10-2.32 (m, 6H) 3.09-3.23 (m, 3H)
3.36-3.44 (m, 4H) 3.60-3.78 (m, 3H) 3.89-3.98 (m, 2H) 6.76 (s, 1H)
7.76 (s, 1H) 8.03 (s, 1H).
Example 63 (Compound 226)
6-(2-(trans-4-aminocyclohexylamino)-5-chloropyridin-4-yl)-N2-((tetrahydro--
2H-pyran-4-yl)methyl)pyrazine-2,3-diamine
##STR00106##
[0561] Step 1. Preparation of
6-bromo-N2-((tetrahydro-2H-pyran-4-yl)methyl)pyrazine-2,3-diamine
[0562] To a scintillation vial containing
3,5-dibromopyrazin-2-amine (500 mg, 1.977 mmol) and TEA (0.551 ml,
3.95 mmol) was added MeCN (6 ml) and
(tetrahydro-2H-pyran-4-yl)methanamine (300 mg, 1.977 mmol). The
homogenous reaction mixture was capped and heated to 80.degree. C.
in a oil bath for 36 hr. The reaction mixture was concentrated to
dryness, diluted with EtOAc and washed with sat NaHCO.sub.3, sat
NaCl. The organic layer was dried Na.sub.2SO.sub.4, filtered and
concentrated. The crude was purified by column chromatography on
silica gel (30% EtOAc/Hexane) to yield
6-bromo-N2-((tetrahydro-2H-pyran-4-yl)methyl)pyrazine-2,3-diamine
(351 mg, 1.222 mmol, 61.8% yield).
Step 2. Preparation of
6-(5-chloro-2-fluoropyridin-4-yl)-N2-((tetrahydro-2H-pyran-4-yl)methyl)py-
razine-2,3-diamine
[0563] To a degassed suspension of
6-bromo-N2-((tetrahydro-2H-pyran-4-yl)methyl)pyrazine-2,3-diamine
(100 mg, 0.348 mmol), Na.sub.2CO.sub.3 (96 mg, 0.905 mmol) and
5-chloro-2-fluoropyridin-4-ylboronic acid (92 mg, 0.522 mmol) in
DME (3 ml) was added PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct
(22.75 mg, 0.028 mmol). The reaction mixture was capped in a flask
and heated to 100.degree. C. for 4 hr an oil bath. The reaction
mixture was diluted with EtOAc and washed with H.sub.2O, sat NaCl.
The organic layer was dried Na2SO4, filtered and concentrated. The
crude was purified by column chromatography on silica gel (100%
EtOAc/Hexane) to yield
6-(5-chloro-2-fluoropyridin-4-yl)-N2-((tetrahydro-2H-pyran-4-yl)methyl)py-
razine-2,3-diamine (34 mg, 0.101 mmol, 28.9% yield).). LCMS (m/z):
338.2 (MH.sup.+), retention time=0.65 min.
Step 3. Preparation of
6-(2-(trans-4-aminocyclohexylamino)-5-chloropyridin-4-yl)-N2-((tetrahydro-
-2H-pyran-4-yl)methyl)pyrazine-2,3-diamine
[0564] To a scintillation vial containing
6-(5-chloro-2-fluoropyridin-4-yl)-N2-((tetrahydro-2H-pyran-4-yl)methyl)py-
razine-2,3-diamine (17 mg, 0.050 mmol) was added DMSO (1.3 ml) and
trans-cyclohexane-1,4-diamine R2 (57.5 mg, 0.503 mmol). The
homogenous reaction mixture was capped and heated to 100.degree. C.
in a oil bath for 16 hr. The reaction mixture was purified by
reverse phase preparative HPLC to yield
6-(2-(trans-4-aminocyclohexylamino)-5-chloropyridin-4-yl)-N2-((tetrahydro-
-2H-pyran-4-yl)methyl)pyrazine-2,3-diamine (13.7 mg, 0.025 mmol,
49.9% yield), LCMS (m/z): 432.1 (MH.sup.+), retention time=0.41 min
as a TFA salt after lypholyzing. 1H NMR (400 MHz, METHANOL-d4) d
ppm 1.30-1.50 (m, 4H) 1.51-1.65 (m, 2H) 1.69-1.78 (m, 2H) 1.93-2.06
(m, 1H) 2.07-2.24 (m, 4H) 3.10-3.19 (m, 1H) 3.36-3.45 (m, 2H) 3.48
(d, J=6.65 Hz, 2H) 3.64-3.75 (m, 1H) 3.96 (dd, J=11.35, 3.13 Hz,
2H) 7.04-7.10 (m, 1H) 7.64 (s, 1H) 8.01 (s, 1H).
Example 64 (Compound 233)
trans-N1-(5-chloro-4-(3-methyl-6-((tetrahydro-2H-pyran-4-yl)methyl)aminopy-
razin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
##STR00107##
[0565] Step 1. Preparation of
6-(5-chloro-2-fluoropyridin-4-yl)-5-methyl-N-((tetrahydro-2H-pyran-4-yl)m-
ethyl)pyrazin-2-amine
[0566] To a degassed suspension of
5-chloro-6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)m-
ethyl)pyrazin-2-amine (10 mg, 0.028 mmol), Na2CO3 (0.036 ml, 2 M,
0.072 mmol) and methylboronic acid (5 mg, 0.084 mmol) in DME (1 ml)
was added PdCl2(dppf).CH2Cl2 adduct (6 mg, 7.35 .mu.mol). The
reaction was capped and heated to 105.degree. C. for 4 hr an oil
bath. The reaction was diluted with EtOAc and washed with H.sub.2O,
sat NaCl. The organic layer was dried Na.sub.2SO.sub.4, filtered
and concentrated. The crude was purified by column chromatography
on silica gel (50% EtOAc/Hexane) to yield
6-(5-chloro-2-fluoropyridin-4-yl)-5-methyl-N-((tetrahydro-2H-pyran--
4-yl)methyl)pyrazin-2-amine (7 mg, 0.021 mmol, 74.2% yield). LCMS
(m/z): 337.2 (MH.sup.+), retention time=0.81 min.
Step 2. Preparation of
trans-N1-(5-chloro-4-(3-methyl-6-((tetrahydro-2H-pyran-4-yl)methyl)aminop-
yrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine
[0567] To a scintillation vial containing
6-(5-chloro-2-fluoropyridin-4-yl)-5-methyl-N-((tetrahydro-2H-pyran-4-yl)m-
ethyl) pyrazin-2-amine (7 mg, 0.021 mmol) was added DMSO and
trans-cyclohexane-1,4-diamine (23.73 mg, 0.208 mmol). The
homogenous reaction mixture was capped and heated to 100.degree. C.
in an oil bath for 4 hr. The crude solution was purified by reverse
phase preparative HPLC to yield
trans-N1-(5-chloro-4-(3-methyl-6-((tetrahydro-2H-pyran-4-yl)methyl)aminop-
yrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine (1.1 mg, 2.018
.mu.mol, 9.71% yield), LCMS (m/z): 431.2 (MH.sup.+), retention
time=0.47 min as a TFA salt after lypholyzing.
Example 65 (Compound 316)
5'-chloro-N6-((6,6-dimethyl-1,4-dioxan-2-yl)methyl)-N2'-(trans-4-((R)-1-me-
thoxypropan-2-yl-amino)cyclohexyl)-2,4'-bipyridine-2',6-diamine
##STR00108##
[0568] Step 1. Preparation of 1-(allyloxy)-2-methylpropan-2-ol
[0569] To allylic alcohol (57.4 mL, 844 mmol) at 0.degree. C. was
added NaH (60% in mineral oil, 2.43 g, 101 mmol). After 20 min
2,2-dimethyloxirane (15 mL, 169 mmol) was added and the solution
was refluxed overnight. Saturated NH.sub.4Cl solution was added and
extracted three times with ether. The organic layers were combined,
dried over Na.sub.2SO.sub.4 and concentrated to remove ether. The
resulting residue was distilled (allylic alcohol was distilled
first then the product was collected at 42 torr, bp 58-60.degree.
C.) to give the product as a colorless oil (12.3 g, 56%). .sup.1H
NMR (400 MHz, CDCl3) .delta. ppm 5.87-5.96 (1H, m), 5.26-5.31 (1H,
m), 5.18-5.21 (1H, m), 4.03-4.05 (2H, m), 3.28 (2H, s), 2.31 (1H,
br s), 1.23, (3H, s), 1.22 (3H, s).
Step 2. Preparation of
2-methyl-1-(oxiran-2-ylmethoxy)propan-2-ol
[0570] 1-(Allyloxy)-2-methylpropan-2-ol (1.50 g, 11.5 mmol) was
dissolved in DCM (50 mL) and cooled to 0.degree. C. mCPBA (77% max,
9.94 g) was added. The suspension was stirred at 0.degree. C. for
6.5 hr. and then saturated NaHCO.sub.3 solution (.about.20 ml) and
Na.sub.2S.sub.2O.sub.3 solution (.about.20 ml) were added. The
resulting mixture was stirred at 0.degree. C. for 15 min and the
two layers were separated. The aqueous layer was extracted twice
with DCM. The organic layers were combined, dried over
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified on a silica gel column (heptane:EtOAc 1:0 to 1:2) to give
the product as a colorless oil (620 mg, 37%). .sup.1H NMR (400 MHz,
CDCl3) .delta. ppm 3.64 (1H, ddd, J=12.0, 5.2, 2.8 Hz), 3.24-3.29
(1H, m), 3.17-3.21 (1H, m), 3.11-3.14 (1H, m), 2.97-3.00 (1H, m),
2.88 (1H, br s), 2.60-2.64 (1H, m), 2.44-2.47 (1H, m), 1.02 (6H,
s).
Step 3. Preparation of (6,6-dimethyl-1,4-dioxan-2-yl)methanol
[0571] 2-methyl-1-(oxiran-2-ylmethoxy)propan-2-ol (620 mg, 4.24
mmol) and 10-CSA (300 mg, 1.29 mmol) were dissolved in DCM (30 mL)
and stirred at ambient temperature for 24 hr. Saturated NaHCO.sub.3
solution was added and the two layers were separated. The aqueous
phase was extracted four times with DCM. The organic layers were
combined, dried over Na.sub.2SO.sub.4 and concentrated. The
resulting residue was purified on a silica gel column
(heptane:EtOAc 1:0 to 1:2) to give the desired product as a
colorless oil (400 mg, 64%). Some starting material was recovered.
.sup.1H NMR (400 MHz, CDCl3) .delta. ppm 3.90-3.96 (1H, m), 3.76
(1H, dd, J=11.2, 2.8 Hz), 3.56 (1H, dd, J=11.6, 4.0 Hz), 3.46-3.50
(2H, m), 3.29 (1H, t, J=11.2 Hz), 3.24 (1H, dd, J=11.6, 1.2 Hz),
2.69 (1H, br s), 1.35 (3H, s), 1.13 (3H, s).
Step 4. Preparation of (6,6-dimethyl-1,4-dioxan-2-yl)methyl
methanesulfonate
[0572] TEA (0.52 mL, 3.74 mmol) and
(6,6-dimethyl-1,4-dioxan-2-yl)methanol (390 mg, 2.67 mmol) were
dissolved in DCM (10 mL). Methanesulfonyl chloride (0.249 mL, 3.20
mmol) was slowly added at 0.degree. C. After the addition was
completed the solution was warmed to ambient temperature and
stirred for 1 hr. Saturated NaHCO.sub.3 solution was added and the
two layers were separated. The aqueous layer was extracted three
times with DCM. The organic layers were combined, dried over
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified on a silica gel column (heptane:EtOAc 4:1 to 1:1) to give
the product as a colorless oil (584 mg, 98%). .sup.1H NMR (400 MHz,
CDCl3) .delta. ppm 4.00-4.09 (3H, m), 3.74 (1H, dd, J=11.2, 2.8
Hz), 3.42 (1H, d, J=11.6 Hz), 3.16-3.23 (2H, m), 2.99 (3H, s), 1.27
(3H, s), 1.05 (3H, s).
Step 5.
6-bromo-N-((6,6-dimethyl-1,4-dioxan-2-yl)methyl)pyridin-2-amine
[0573] 6-Bromopyridin-2-amine (722 mg, 4.17 mmol) was dissolved in
8 mL of anhydrous DMF and cooled to 0.degree. C. NaH (60% in
mineral oil, 195 mg, 4.87 mmol) was added. After 10 min the
solution was warmed to ambient temperature and stirred for 45 min
until bubbling ceased. The solution was cooled to 0.degree. C.
again and (6,6-dimethyl-1,4-dioxan-2-yl)methyl methanesulfonate
(520 mg, 2.32 mmol) in 2 mL of DMF was added. After the addition
was completed the solution was warmed to ambient temperature and
stirred overnight. It was diluted with EtOAc and washed four times
with water. The aqueous layers were combined and extracted once
with EtOAc. The organic layers were combined, dried over Na2SO4 and
concentrated. The resulting residue was purified on prep HPLC and
the collected fractions were combined, concentrated, basified with
Na2CO3 and extracted with EtOAc three times. The organic layers
were combined, dried over Na2SO4 and concentrated to give the
product as a light yellow oil (270 mg, 39%). LC-MS (m/z):
301.0/303.0 (M+H), retention time=0.86 min.
Example 66 (Compound 307)
5'-chloro-N6-((5,5-dimethyl-1,4-dioxan-2-yl)methyl)-N2'-(trans-4-((R)-1-me-
thoxypropan-2-yl-amino)cyclohexyl)-2,4'-bipyridine-2',6-diamine
##STR00109##
[0574] Step 1. Preparation of 2-(allyloxy)-2-methylpropan-1-ol
[0575] 2,2-Dimethyloxirane (15.0 mL, 169 mmol) was dissolved in
allylic alcohol (57.4 mL) and cooled to 0.degree. C. Perchloric
acid (70%, 7.26 mL, 84 mmol) was slowly added. The solution was
then warmed to ambient temperature and stirred for 1.5 hr.
Saturated NaHCO.sub.3 solution was added and extracted three times
with ether. The organic layers were combined, dried over
Na.sub.2SO.sub.4 and concentrated to remove ether. The resulting
residue was distilled (allylic alcohol was distilled first then the
product was collected at 38 torr, bp 74-76.degree. C.) to give the
product as a colorless oil (9.70 g, 44%). .sup.1H NMR (400 MHz,
CDCl3) .delta. ppm 5.87-5.97 (1H, m), 5.25-5.31 (1H, m), 5.12-5.16
(1H, m), 3.92-3.94 (2H, m), 3.45 (2H, m), 1.19 (6H, s).
Step 2. Preparation of
2-methyl-2-(oxiran-2-ylmethoxy)propan-1-ol
[0576] 2-(allyloxy)-2-methylpropan-1-ol (2.37 g, 18.2 mmol) was
dissolved in DCM (70 mL) and cooled to 0.degree. C. mCPBA (77% max,
15.71 g) was added. The suspension was stirred at 0.degree. C. for
6.5 hr before saturated NaHCO.sub.3 solution and Na.sub.2S203
solution were added. It was stirred at 0.degree. C. for 15 min and
the two layers were separated. The aqueous layer was extracted
twice with DCM. The organic layers were combined, dried over Na2SO4
and concentrated. The resulting residue was purified on a silica
gel column (heptane:EtOAc 1:0 to 1:2) to give the product as a
colorless oil (910 mg, 34%). .sup.1H NMR (400 MHz, CDCl3) .delta.
ppm 3.65 (1H, dd, J=11.2, 2.8 Hz), 3.47 (1H, br s), 3.31-3.41 (3H,
m), 3.07-3.09 (1H, m), 2.74 (1H, t, J=4.8 Hz), 2.63-2.65 (1H, m),
1.12 (6H, s).
Step 3. Preparation of (5,5-dimethyl-1,4-dioxan-2-yl)methanol
[0577] 2-Methyl-2-(oxiran-2-ylmethoxy)propan-1-ol (870 mg, 5.95
mmol) and 10-CSA (207 mg, 15%) were dissolved in DCM (70 mL) and
stirred at ambient temperature for 24 hr. More 10-CSA (100 mg) was
added and the solution was stirred overnight. Saturated NaHCO3
solution was added. The two layers were separated and the aqueous
layer was extracted twice with DCM. The organic layers were
combined, dried over Na2SO4 and concentrated to give the product as
a colorless oil (750 mg, 86%). .sup.1H NMR (400 MHz, CDCl3) .delta.
ppm 3.69-3.74 (1H, m), 3.52-3.64 (5H, m), 3.43 (1H, dd, J=11.6, 0.8
Hz), 2.57 (1H, br s), 1.32 (3H, s), 1.13 (3H, s).
Step 4. Preparation of (5,5-dimethyl-1,4-dioxan-2-yl)methyl
methanesulfonate
[0578] (5,5-Dimethyl-1,4-dioxan-2-yl)methanol (740 mg, 5.06 mmol)
and TEA (0.988 mL, 7.09 mmol) were dissolved in DCM (20 mL). At
0.degree. C. MsCl (0.473 mL, 6.07 mmol) was added dropwise. After
the addition the solution was warmed to ambient temperature and
stirred for 1 hr. Saturated NaHCO3 solution was added and the two
layers were separated. The aqueous layer was extracted three times
with DCM. The organic layers were combined, dried over Na2SO4 and
concentrated. The resulting residue was purified on a silica gel
column (heptane:EtOAc 4:1 to 1:1) to give the product as a
colorless oil (805 mg, 71%). .sup.1H NMR (400 MHz, CDCl3) .delta.
ppm 4.18-4.19 (2H, m), 3.71-3.76 (1H, m), 3.66 (1H, t, J=10.8 Hz),
3.52-3.57 (2H, m), 3.37 (1H, d, J=11.6 Hz), 3.03 (3H, s), 1.28 (3H,
s), 1.09 (3H, s).
Step 5. Preparation of
6-bromo-N-((5,5-dimethyl-1,4-dioxan-2-yl)methyl)pyridin-2-amine
[0579] 6-Bromopyridin-2-amine (771 mg, 4.46 mmol) was dissolved in
10 mL of anhydrous DMF and cooled to 0.degree. C. NaH (60% in
mineral oil, 214 mg, 5.35 mmol) was added. After 10 min the
solution was warmed to ambient temperature and stirred for 15 min
until bubbling ceased, to give a dark green solution.
(5,5-Dimethyl-1,4-dioxan-2-yl)methyl methanesulfonate (500 mg, 2.23
mmol) in 2 mL of DMF was added. After the addition was completed
the solution was stirred at ambient temperature for 20 min, then
heated at 60.degree. C. for 1.5 hr. It was diluted with EtOAc and
washed four times with water. The aqueous layers were combined and
extracted once with EtOAc. The organic layers were combined, dried
over Na2SO4 and concentrated. The resulting residue was purified on
a silica gel column (heptane:EtOAc 1:0 to 1:1) to give the product
contaminated with the starting aminopyridine. Another purification
on silica gel column (DCM:ether 20:1) gave the pure product (306
mg, 46%). LC-MS (m/z): 301.0/303.0 (M+H), retention time=0.89
min.
Example 67 (Compound 291)
Synthesis of
5'-chloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-((t-
etrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00110##
[0581] Step 1. To sodium hydride (0.488 g, 12.21 mmol) in 5 mL of
THF was added via syringe (S)-(+)-3-methoxy-2-propanol (1.000 ml,
11.10 mmol) in 25 mL of THF at ambient temperature. The mixture was
stirred for 20 min. and followed by addition of p-toluenesulfonyl
chloride (2.327 g, 12.21 mmol). The white cloudy solution was
stirred at ambient temperature for 18 hrs. The reaction mixture was
diluted with saturated aq. NaHCO.sub.3 and extracted with EtOAc.
The organic extracts were combined, washed with brine, dried with
sodium sulfate and concentrated in vacuo to give 2 g of colorless
liquid. The crude mixture was purified by Analogix system (silica
gel column 40 g, gradient: 100% n-heptane to 30% EtOAc in Heptane;
30 min.). The pure fractions were concentrated in vacuo to give
1.22 g of colorless oil. LC-MS (m/z): 245 (M+H), retention
time=0.83 min.
[0582] Step 2. To the tosylate obtained from step 1 (0.6 g, 2.45
mmol) in DMSO (6 ml) at ambient temperature was added
trans-cyclohexane-1,4-diamine (0.84 g, 7.37 mmol). The light brown
mixture was heated to 99-C in a capped glass vial for 1 hr. LC/MS
showed nearly complete consumption of the starting material. The
mixture was diluted with water and extracted with DCM. The organic
extracts were combined, washed with brine, dried with sodium
sulfate and concentrated in vacuo to give 0.39 g of light brown
liquid. This was used in the next step without further
purification. LC-MS (m/z): 187 (M+H), Retention time=0.14 min.
[0583] Step 3. A mixture of Intermediate G (60 mg, 0.168 mmol), the
above cyclohexadiamine (100 mg, 0.537 mmol) and 2,6-LUTIDINE (0.039
ml, 0.0.337 mmol) in DMSO (1 ml) was heated in a capped vial on a
heating block for 18 hrs. LC/MS showed containing about 50%
product. The reaction mixture was purified by HPLC (ACN in water
with gradient 10%-50% in 16 minutes) and lyophilized to give 25 mg
of light yellow powder. LC-MS (m/z): 522/524 (M+H), retention
time=0.62 min. .sup.1H NMR (400 MHz, CDCl3) .delta. ppm 1.24-1.47
(m, 5H) 1.50-1.79 (m, 2H) 1.79-2.01 (m, 4H) 2.11-2.31 (m, 4H)
3.16-3.26 (m, 2H) 3.28-3.45 (m, 5H) 3.45-3.66 (m, 4H) 6.82 (d,
J=9.39 Hz, 1H) 7.05 (br. s., 1H) 7.59 (s, 1H) 7.78 (d, J=9.39 Hz,
1H) 7.95 (s, 1H) 8.76 (br. s., 1H)
Example 68 (Compound 197)
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-((methylamino)methyl)cyclohexyl-
)-2,4'-bipyridine-2',6-diamine
##STR00111##
[0585] Step 1. Preparation of
2'-chloro-5'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine: To a
solution of 6-bromo-N-(3-fluorobenzyl)pyridin-2-amine (636 mg,
2.262 mmol) and 2-chloro-5-fluoropyridin-4-yl-boronic acid (555 mg,
3.17 mmol) in DME (4 ml) and 2M Na.sub.2CO.sub.3 aq (2 ml) was
added PdCl.sub.2(dppf). CH.sub.2Cl.sub.2 adduct (92 mg, 0.113
mmol). This was then heated at 110.degree. C. for 16 hours. The
reaction mixture was allowed to cool and then the DME was
evaporated under reduced pressure. The resulting residue was
partitioned between EtOAc and water. The organics were combined,
then washed with H.sub.2O (.times.3), saturated aq. brine
(.times.3), then dried (Na.sub.2SO.sub.4), filtered and evaporated
under reduced pressure. The resulting residue was purified by flash
column chromatography (silica gel; 20% EtOAc/hexane) to give
2'-chloro-5'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine (84
mg).
[0586] Step 2. Preparation of
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-((methylamino)methyl)cyclohexy-
l)-2,4'-bipyridine-2',6-diamine: To a scintillation vial were added
2'-chloro-5'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine (34
mg, 0.102 mmol), trans-cyclohexane-1,4-diamine (52.7 mg, 0.461
mmol),
1,3-bis(2,6-di-isopropylphenyl)imidazol-2-ylidene(1,4-naphthoquinone)pall-
adium(0) (13.39 mg, 10.25 mol), KOH (51.8 mg, 0.922 mmol) and
Dioxane (0.6 mL). The resulting mixture was stirred with heating at
70.degree. C. for 16 h and then concentrated in vacuo. The
resulting residue was dissolved in EtOAc and washed with H.sub.2O
(.times.2) followed by saturated brine (.times.2), then dried
(Na.sub.2SO.sub.4), filtered and evaporated under reduced pressure.
The resulting residue was purified by reverse phase preparative
HPLC and then lyophallized to yield
5'-chloro-N6-(3-fluorobenzyl)-N2'-(trans-4-((methylamino)methyl)cyclohexy-
l)-2,4'-bipyridine-2',6-diamine (7.9 mg), LCMS (m/z): 410.3
(MH.sup.+), retention time=0.60 min as a TFA salt. .sup.1H-NMR (400
MHz, METHANOL-d4, 25.degree. C.) 1.40-1.70 (m, 4H) 2.05-2.25 (m,
4H) 3.10-3.25 (m, 1H) 3.55-3.64 (m, 1H) 4.57 (s, 2H) 6.76 (d, J=8.4
Hz, 1H) 6.93-7.00 (m, 1H) 7.11 (d, J=10.4 Hz, 1H) 7.20 (m, 2H)
7.28-7.36 (m, 1H) 7.52 (d, J=6.4 Hz, 1H) 7.61 (t, J=8.0 Hz, 1H)
7.96 (d, J=4.8 Hz, 1H).
Example 69 (Compound 180)
N2'-(trans-4-aminocyclohexyl)-N6-(3-fluorobenzyl)-5'-methoxy-2,4'-bipyridi-
ne-2',6-diamine
[0587] Step 1. Preparation of
2'-chloro-N-(3-fluorobenzyl)-5'-methoxy-2,4'-bipyridin-6-amine: To
a solution of 6-bromo-N-(3-fluorobenzyl)pyridin-2-amine (555 mg,
1.974 mmol) and 2-chloro-5-methoxypyridin-4-ylboronic acid (518 mg,
2.76 mmol) in DME (4 ml) and 2M Na.sub.2CO.sub.3 aq (2 ml) was
added PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (81 mg, 0.099 mmol).
This was then heated at 110.degree. C. for 5 h. The reaction
mixture was allowed to cool and then the DME was evaporated under
reduced pressure. The resulting residue was partitioned between
EtOAc and water. The organics were combined, then washed with
H.sub.2O (.times.3), saturated aq. brine (.times.3), then dried
(Na.sub.2SO.sub.4), filtered and evaporated under reduced pressure.
The resulting residue was purified by flash column chromatography
(silica gel; 15% to 25% EtOAc/hexane) to give
2'-chloro-N-(3-fluorobenzyl)-5'-methoxy-2,4'-bipyridin-6-amine (53
mg).
[0588] Step 2. Preparation of
N2'-(trans-4-aminocyclohexyl)-N6-(3-fluorobenzyl)-5'-methoxy-2,4'-bipyrid-
ine-2',6-diamine: To a scintillation vial was added
2'-chloro-N-(3-fluorobenzyl)-5'-methoxy-2,4'-bipyridin-6-amine (30
mg, 0.087 mmol), trans-cyclohexane-1,4-diamine (45 mg, 0.394 mmol),
1,3-bis(2,6-di-isopropylphenyl)imidazol-2-ylidene(1,4-naphthoquinone)pall-
adium(0) (11.4 mg, 8.73 .mu.mol), KOH (45 mg, 0.802 mmol) and
Dioxane (0.3 mL). The resulting mixture was stirred at 100.degree.
C. for 18 h. The mixture was concentrated in vacuo and then diluted
with water. The resultant solid was filtered and washed with water
(.times.3). The solid was then purified by reverse phase
preparative HPLC and then lyophallized to yield
N2'-(trans-4-aminocyclohexyl)-N6-(3-fluorobenzyl)-5'-methoxy-2,4-
'-bipyridine-2',6-diamine (6.5 mg), LCMS (m/z): 422.3 (MH.sup.+),
retention time=0.54 min as a TFA salt. .sup.1H-NMR (400 MHz,
METHANOL-d4, 25.degree. C.) 1.40-1.66 (m, 4H) 2.05-2.25 (m, 4H)
3.10-3.25 (m, 1H) 3.55-3.64 (m, 1H) 3.86 (s, 3H) 4.57 (s, 2H) 6.69
(d, J=8.4 Hz, 1H) 6.92-7.00 (m, 1H) 7.10 (d, J=10.0 Hz, 1H) 7.17
(d, J=7.6 Hz, 1H) 7.28-7.33 (m, 2H) 7.48-7.52 (m, 2H) 7.53-7.58 (m,
1H).
Example 70 (Compound 211)
N2'-(trans-4-aminocyclohexyl)-N6-(3-fluorobenzyl)-5'-methyl-2,4'-bipyridin-
e-2',6-diamine
##STR00112##
[0590] Step 1. Preparation of
2'-fluoro-N-(3-fluorobenzyl)-5'-methyl-2,4'-bipyridin-6-amine: To a
solution of 6-bromo-N-(3-fluorobenzyl)pyridin-2-amine (85 mg, 0.302
mmol) and
2-fluoro-5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyri-
dine (102 mg, 0.430 mmol) in DME (2 mL) and 2M Na.sub.2CO.sub.3 aq
(1 mL) was added PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct (21 mg,
0.026 mmol). This was then heated at 110.degree. C. for 16 h. The
reaction mixture was allowed to cool and then the DME was
evaporated under reduced pressure. The resulting residue was
partitioned between EtOAc and water. The organics were combined,
then washed with H.sub.2O (.times.3), saturated aq. brine
(.times.3), then dried (Na.sub.2SO.sub.4), filtered and evaporated
under reduced pressure. The resulting residue was purified by flash
column chromatography (silica gel; 15% to 25% EtOAc/hexane) to give
2'-fluoro-N-(3-fluorobenzyl)-5'-methyl-2,4'-bipyridin-6-amine (43
mg).
[0591] Step 2. Preparation of
N2'-(trans-4-aminocyclohexyl)-N6-(3-fluorobenzyl)-5'-methyl-2,4'-bipyridi-
ne-2',6-diamine: To a solution of
2'-fluoro-N-(3-fluorobenzyl)-5'-methyl-2,4'-bipyridin-6-amine (18
mg, 0.058 mmol) and trans-cyclohexane-1,4-diamine (39.6 mg, 0.347
mmol), in NMP (0.3 mL) was added DIPEA (20 .mu.L, 0.115 mmol). The
mixture was heated at 130.degree. C. for 48 h. The mixture was
allowed to cool then diluted with water and then extracted with
EtOAc (.times.3). The combined organics were washed with saturated
brine (.times.2), then dried (Na.sub.2SO.sub.4), filtered and
evaporated under reduced pressure. The resulting residue was
purified by reverse phase preparative HPLC and then lyophallized to
yield
N2'-(trans-4-aminocyclohexyl)-N6-(3-fluorobenzyl)-5'-methyl-2,4'-bipyridi-
ne-2',6-diamine (4.2 mg), LCMS (m/z): 406.3 (MH.sup.+), retention
time=0.53 min as a TFA salt.
Example 71 (Compound 280)
Racemic
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(-
tetrahydrofuran-3-yl-amino)cyclohexyl)-2,4'-bipyridine-2',6-diamine
##STR00113##
[0592] Step 1. Preparation of racemic benzyl
trans-4-(tetrahydrofuran-3-yl-amino)cyclohexylcarbamate
[0593] To a stirred solution of benzyl
trans-4-aminocyclohexylcarbamate (396 mg, 1.595 mmol) in
CH.sub.2Cl.sub.2 (9 ml) was added dihydrofuran-3(2H)-one (151 mg,
1.754 mmol) followed by acetic acid (150 .mu.L, 2.62 mmol) and
sodium triacetoxyborohydride (439 mg, 2.073 mmol) under Argon.
Stirred at 25.degree. C. for 16 h, then concentrated in vacuo. The
resulting residue was partitioned between EtOAc and 1M NaOH. The
organics were combined, then washed with 1M NaOH (.times.2), water
(.times.2), saturated brine (.times.2), then dried
(Na.sub.2SO.sub.4), filtered and evaporated under reduced pressure
to give racemic benzyl
trans-4-(tetrahydrofuran-3-yl-amino)cyclohexylcarbamate (495 mg).
The resulting residue was used in next step without further
purification.
Step 2. Preparation of racemic tert-butyl
trans-4-aminocyclohexyl(tetrahydrofuran-3-yl)carbamate
[0594] To a stirred solution of racemic benzyl
trans-4-(tetrahydrofuran-3-yl-amino)cyclohexylcarbamate (495 mg,
1.555 mmol) in CH.sub.2Cl.sub.2 (5 ml) was added BOC-Anhydride
(0.397 ml, 1.710 mmol) and the resulting mixture was stirred at
25.degree. C. under Argon for 21 hours. The mixture was evaporated
under reduced pressure and purified by flash column chromatography
(silica gel; 15% to 25% EtOAc/hexane). A solution of the resultant
Boc protected intermediate (135 mg, 0.323 mmol) in MeOH (5 mL) was
hydrogenated under an atmosphere of hydrogen in the presence of 10%
Pd/C (24 mg, 0.226 mmol) for 18 h. The mixture was then filtered
through Celite and the filtrate evaporated under reduced pressure
to give racemic tert-butyl
trans-4-aminocyclohexyl(tetrahydrofuran-3-yl)carbamate (100 mg).
The resulting residue was used in next step without further
purification
Step 3. Preparation of racemic
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(tetrahy-
drofuran-3-yl-amino)cyclohexyl)-2,4'-bipyridine-2',6-diamine
[0595] To a scintillation vial was added
3,5'-dichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
in-6-amine (25 mg, 0.070 mmol), racemic tert-butyl
trans-4-aminocyclohexyl(tetrahydrofuran-3-yl)carbamate (21.95 mg,
0.077 mmol), DIPEA (24.51 .mu.l, 0.140 mmol) and NMP (0.2 ml). This
was heated at 110.degree. C. for 48 h. The mixture was diluted with
EtOAc and washed with water (.times.2), saturated brine (.times.2),
then dried (Na.sub.2SO.sub.4), filtered and evaporated under
reduced pressure. The resulting residue was dissolved in
CH.sub.2Cl.sub.2 (0.4 mL) and treated with TFA (100 .mu.l, 1.298
mmol). After 30 minutes, the mixture was concentrated in vacuo and
the resulting residue was purified by reverse phase preparative
HPLC and then lyophallized to yield racemic
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(tetrahy-
drofuran-3-yl-amino)cyclohexyl)-2,4'-bipyridine-2',6-diamine (10.8
mg), LCMS (m/z): 520.1/522.0 (bis-chloro isotopic signature for
MH.sup.+), retention time=0.59 min as a TFA salt.
Example 72 (Compound 320)
3,5'-dichloro-N2'-(trans-4-(((R)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)ami-
nocyclohexyl)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2'6-di-
amine
##STR00114##
[0597] To a stirred solution of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine (68 mg, 0.151 mmol) in DMF
(0.2 ml) was added DIPEA (80 .mu.L, 0.458 mmol) followed by
(S)-(2,2-dimethyl-1,3-dioxolan-4-yl)methyl 4-methylbenzenesulfonate
(42 mg, 0.147 mmol). The mixture was heated at 75.degree. C. for 19
hours. The mixture was allowed to cool, then diluted with water and
then extracted with EtOAc (.times.3). The organics were combined
then washed with water (.times.2), saturated brine (.times.2), then
dried (Na.sub.2SO.sub.4), filtered and evaporated under reduced
pressure. The resulting residue was purified by reverse phase prep
HPLC and lyaphllized to yield
3,5'-dichloro-N2'-(trans-4-(((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-
methyl)aminocyclohexyl)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyrid-
ine-2',6-diamine (4.4 mg), LCMS (m/z): 564.4/566.3 (bis-chloro
isotopic signature for MH.sup.+) retention time=0.65 min as a TFA
salt.
Example 73 (Compounds 323 and 327)
[0598]
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohex-
yl)-N6-(((S)-tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2',6-diamine
and
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl-
)-N6-(((R)-tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00115##
[0599] Step 1. Preparation of racemic
(tetrahydro-2H-pyran-3-yl)methyl 4-methylbenzenesulfonate: To a
stirred solution of (tetrahydro-2H-pyran-3-yl)methanol (1.0 g, 8.61
mmol) and DMAP (0.053 g, 0.430 mmol) in CH.sub.2Cl.sub.2 (5.0 mL)
and pyridine (6.96 mL, 86 mmol) was added Tosyl-Cl (1.805 g, 9.47
mmol). (11:23 am). After 16 h the mixture was evaporated under
reduced pressure and the resulting residue partitioned between
EtOAc and water. The organics were separated, then washed with 0.1M
HCl (.times.3), H.sub.2O (.times.1), saturated aq. NaHCO.sub.3
(.times.2), H.sub.2O (.times.1), saturated brine (.times.1), then
dried (Na.sub.2SO.sub.4), filtered and evaporated under reduced
pressure to give racemic (tetrahydro-2H-pyran-3-yl)methyl
4-methylbenzenesulfonate (2.034 g). The resulting residue was used
in next step without further purification.
[0600] Step 2. Preparation of racemic tert-butyl
6-bromo-5-chloropyridin-2-yl((tetrahydro-2H-pyran-3-yl)methyl)carbamate:
To a cooled (0.degree. C.), stirred solution of tert-butyl
6-bromo-5-chloropyridin-2-ylcarbamate (1.00 g, 3.25 mmol) in DMF
(13.0 mL) was added 60% dispersion NaH (0.156 g, 3.90 mmol) under
Argon. Stirred at 0.degree. C. for 30 mins then added racemic
(tetrahydro-2H-pyran-3-yl)methyl 4-methylbenzenesulfonate (1.143 g,
4.23 mmol). The mixture was then allowed to warm to 25.degree. C.
and stirring continued for 19 h. The reaction mixture then was
diluted with saturated NH.sub.4Cl and then extracted with EtOAc
(.times.3). Organics washed with water (.times.2), saturated brine
(.times.2), then dried (Na.sub.2SO.sub.4), filtered and evaporated
under reduced pressure. The resulting residue was purified by flash
column chromatography (silica gel; 5% to 15% EtOAc/heptanes) to
give racemic tert-butyl
6-bromo-5-chloropyridin-2-yl((tetrahydro-2H-pyran-3-yl)methyl)carbamate
(938 mg).
[0601] Step 3. Preparation of tert-butyl
3,5'-dichloro-2'-fluoro-2,4'-bipyridin-6-yl((tetrahydro-2H-pyran-3-yl)met-
hyl)carbamate: To a scintillation vial was added racemic tert-butyl
6-bromo-5-chloropyridin-2-yl((tetrahydro-2H-pyran-3-yl)methyl)carbamate
(832 mg, 2.051 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid
(719 mg, 4.10 mmol) and PdCl.sub.2(dppf).CH.sub.2Cl.sub.2 adduct
(167 mg, 0.205 mmol) followed by DME (3 mL) and 2M Na.sub.2CO.sub.3
aq (2 mL). The mixture was heated at 90.degree. C. for 20 h, then
allowed to cool and added water and then extracted with EtOAc
(.times.3). The organics were washed with water (.times.2),
saturated brine (.times.2), then dried (Na.sub.2SO.sub.4), filtered
and evaporated under reduced pressure. The resulting residue was
purified by flash column chromatography (silica gel; 5% to 15%
EtOAc/heptanes) to give racemic tert-butyl
3,5'-dichloro-2'-fluoro-2,4'-bipyridin-6-yl((tetrahydro-2H-pyran-3-yl)met-
hyl)carbamate (374 mg)
[0602] Step 4. Preparation of
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-(((S)-tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2',6-diamine
and
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-(((R)-tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2',6-diamine:
[0603] To a scintillation vial was added racemic tert-butyl
3,5'-dichloro-2'-fluoro-2,4'-bipyridin-6-yl((tetrahydro-2H-pyran-3-yl)met-
hyl)carbamate (114 mg, 0.250 mmol),
trans-N1-((R)-1-methoxypropan-2-yl)cyclohexane-1,4-diamine (70 mg,
0.376 mmol) and DIPEA (0.088 ml, 0.501 mmol) followed by NMP (0.1
ml). The mixture was heated at 110.degree. C. for 60 hr then
concentrated in vacuo. The resulting residue was purified by
reverse phase prep HPLC and lyapholized. The resulting white solid
was free based by dissolving in EtOAc and then washing with 1M NaOH
(.times.3), water (.times.2), saturated brine (.times.2), then
dried (Na.sub.2SO.sub.4), filtered and evaporated under reduced
pressure. The resulting residue was then purified by chiral
separation chromatography to yield
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-(((S)-tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2'6-diamine
(mg), LCMS (m/z): 522.1/523.9 (MH.sup.+), t.sub.R=0.675 min. and
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-(((R)-tetrahydro-2H-pyran-3-yl)methyl)-2,4'-bipyridine-2',6-diamine
(mg) LCMS 522.1/523.9 (m/z): (MH.sup.+), retention time=0.675
min.
Example 74 (Compounds 321 and 325)
[0604]
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohex-
yl)-N6-(((S)-tetrahydro-2H-pyran-2-yl)methyl)-2,4'-bipyridine-2',6-diamine
and
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl-
)-N6-(((R)-tetrahydro-2H-pyran-2-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00116##
[0605] The compounds were prepared according to Example 73, except
using tetrahydro-2H-pyran-2-yl)methanol to give
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-(((S)-tetrahydro-2H-pyran-2-yl)methyl)-2,4'-bipyridine-2',6-diamine
LCMS (m/z): 522.1/524.1 (MH.sup.+), retention time=0.708 min and
3,5'-dichloro-N2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexyl)-N6-
-(((R)-tetrahydro-2H-pyran-2-yl)methyl)-2,4'-bipyridine-2',6-diamine
LCMS (m/z): 522.1/524.1 (MW.sup.+), retention time=0.708 min.
Example 75 (Compound 208)
trans-4-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyrazin-2-yl-
)pyridin-2-yl-amino)cyclohexanol
##STR00117##
[0606] Step 1: Preparation of
6-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
[0607] A mixture of 2,6-dichloropyrazine (950 mg, 6.38 mmol), DMSO
(14 ml), TEA (1.067 ml, 7.65 mmol) and
(tetrahydro-2H-pyran-4-yl)methanamine (771 mg, 6.70 mmol) was
stirred at 75.degree. C. for 6 hours, and the reaction progress was
followed by LCMS. The crude reaction mixture was cooled to ambient
temperature, diluted with 300 ml of ethyl acetate, washed with 1M
NaOH soln. (1.times.), water (1.times.), saturated salt soln.
(1.times.), dried with sodium sulfate, filtered, and concentrated
to constant mass, giving 1185 mg of titled compound as free base,
used without further purification. LCMS (m/z): 228.0 (MH+),
retention time=0.73 min.
Step 2. Preparation of
6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)methyl)pyr-
azin-2-amine
[0608] A mixture of
6-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine (1390
mg, 6.10 mmol), 5-chloro-2-fluoropyridin-4-ylboronic acid (2141 mg,
12.21 mmol), PdCl2(dppf).CH2Cl2 adduct (399 mg, 0.488 mmol), DME
(24 ml) and 2M sodium carbonate (9.16 ml, 18.31 mmol) was stirred
at 110-115.degree. C. for 90 min and the reaction progress was
followed by LCMS. The reaction mixture was cooled, 30 ml of ethyl
acetate and 20 ml of methanol were added, filtered and concentrated
to crude product. The crude was purified by silica gel
chromatography using 80 g column eluting with 20-75% ethyl acetate
in heptane. The desired fractions were concentrated to constant
mass, giving 980 mg of titled compound as free base. LCMS (m/z):
323.0 (MH+), retention time=0.81 min.
Step 3. Preparation of
trans-4-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyrazin-2-y-
l)pyridin-2-yl-amino)cyclohexanol
[0609] A mixture of
6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)methyl)pyr-
azin-2-amine (375 mg, 1.162 mmol), DMSO (3.5 ml) and
trans-4-aminocyclohexanol (1204 mg, 10.46 mmol) was stirred at
100.degree. C. for 18 hours and the progress was followed by LCMS.
The reaction mixture was let cool, added 300 ml of ethyl acetate,
washed with saturated sodium bicarbonate solution (3.times.), water
(2.times.), saturated salt solution (1.times.), dried with sodium
sulfate, filtered and concentrated to crude solid. The crude
material was purified by silica gel chromatography using 40 g
column, eluting slowly from (80% ethyl acetate 20% heptane with 2%
MeOH) to 100% ethyl acetate with 2% MeOH. The desired fractions are
concentrated to a constant mass, lyapholized from 1:1 ACN/water
(does not fully dissolve), re-lyophilized from 80 ml of (60/40)
ACN/water solution with sonicating to dissolve solid, giving 270 mg
of title compound as free base. LCMS (m/z): 418.3 (MH+), retention
time=0.52 min.; 1H NMR (300 MHz, METHANOL-d4, 25.degree. C.)
.delta. ppm 1.19-1.55 (m, 6H) 1.71 (d, J=12.89 Hz, 2H) 1.85-2.15
(m, 5H) 3.28-3.32 (dMeOH, 2H App.) 3.40 (td, J=11.72, 1.76 Hz, 2H)
3.50-3.73 (m 2H) 3.94 (dd, J=11.28, 3.08 Hz, 2H) 6.66 (s, 1H) 7.86
(s, 2H) 7.99 (s, 1H)
Example 76 (Compound 215 and 216)
1-((R)-3-((2'-(trans-4-aminocyclohexylamino)-5'-chloro-2,4'-bipyridin-6-yl-
-amino)methyl)piperidin-1-yl)ethanone and
1-((R)-3-((2'-(trans-4-aminocyclohexylamino)-2,4'-bipyridin-6-yl-amino)me-
thyl)piperidin-1-yl)ethanone
##STR00118##
[0610] Step 1. Preparation of (R)-tert-butyl
3-((2'-(trans-4-aminocyclohexylamino)-5'-chloro-2,4'-bipyridin-6-yl-amino-
)methyl)piperidine-1-carboxylate
[0611] A mixture of
trans-N1-(5'-chloro-6-fluoro-2,4'-bipyridin-2'-yl)cyclohexane-1,4-diamine
(Example 1a, step 2) (50 mg, 0.156 mmol), DMSO (0.75 ml),
(R)-tert-butyl 3-(aminomethyl)piperidine-1-carboxylate (167 mg,
0.779 mmol) and TEA (0.033 ml, 0.234 mmol) was stirred at
100-105.degree. C. for 40 hours and the reaction progress was
followed by LCMS. The reaction mixture was let cool, added 0.75 ml
of DMSO, filtered and purified by prep LC, and lyapholized to yield
36 mg of titled compound as a TFA salt. LCMS (m/z): 515.4 (MH+),
retention time=0.64 min.;
Step 2. Preparation of benzyl
trans-4-(5'-chloro-6-((S)-piperidin-3-yl-methylamino)-2,4'-bipyridin-2'-y-
l-amino)cyclohexylcarbamate
[0612] A mixture of (R)-tert-butyl
3-((2'-(trans-4-aminocyclohexylamino)-5'-chloro-2,4'-bipyridin-6-yl-amino-
)methyl)piperidine-1-carboxylate (36 mg, 0.070 mmol), DCM (1.2 ml),
TEA (0.019 ml, 0.140 mmol) and benzyl 2,5-dioxopyrrolidin-1-yl
carbonate (26.1 mg, 0.105 mmol) was stirred at ambient temperature
for 2 hours and the reaction progress was followed by LCMS. To this
crude reaction mixture was added 25 ml of ethyl acetate, washed
with 2M sodium carbonate, water (2.times.) and saturated salt
solution (1.times.), dried with sodium sulfate, filtered,
concentrated to crude intermediate. To the crude intermediate was
added 4M HCl in Dioxane (2 ml, 8.00 mmol) and stirred at ambient
temperature for 1 hour. The crude reaction mixture was concentrated
to constant mass, dissolved in 1 ml of DMSO and purified by prep
LC. After lypohilization, 15 mg of the title compound, was obtained
as a TFA salt. LCMS (m/z): 549.4 (MH+), retention time=0.67
min.
Step 3. Preparation of
1-((R)-3-((2'-(trans-4-aminocyclohexylamino)-5'-chloro-2,4'-bipyridin-6-y-
l-amino)methyl)piperidin-1-yl)ethanone and
1-((R)-3-((2'-(trans-4-aminocyclohexylamino)-2,4'-bipyridin-6-yl-amino)me-
thyl)piperidin-1-yl)ethanone
[0613] A mixture of benzyl
trans-4-(5'-chloro-6-((S)-piperidin-3-yl-methylamino)-2,4'-bipyridin-2'-y-
l-amino)cyclohexylcarbamate (15 mg, 0.027 mmol), DCM (2 mL), TEA
(0.011 mL, 0.082 mmol) and acetic anhydride (3.09 .mu.L, 0.033
mmol) was stirred at ambient temperature for 2 hours and the
reaction progress was followed by LCMS. The solvent was
concentrated off. The reaction mixture flask was flushed with
argon, 10% palladium on activated carbon (5 mg, 4.70 .mu.mol) was
added and followed by careful addition of MeOH (0.8 mL). The
resulting mixture was stirred under hydrogen for 45 minutes at
ambient temperature and monitored by LCMS. To the crude reaction
mixture was added 2 ml of DCM, filtered and the solvent was
concentrated off. The resulting residue was dissolved in 1.0 ml of
DMSO, filtered and purified by prep HPLC to give two fractions
corresponding to the two title compounds respectively. After
lypohilization, 4.0 mg of
1-((R)-3-((2'-(trans-4-aminocyclohexylamino)-5'-chloro-2,4'-bipyridin-6-y-
l-amino)methyl)piperidin-1-yl)ethanone, was obtained as a TFA salt.
LCMS (m/z): 457.2 (MH.sup.+), retention time=0.46 min. In addition,
1.0 mg of
1-((R)-3-((2'-(trans-4-aminocyclohexylamino)-2,4'-bipyridin-6-yl-amino)me-
thyl)piperidin-1-yl)ethanone, as TFA salt was also obtained. LCMS
(m/z): 423.2 (MH+), retention time=0.45 min. This reaction yielded
two products which are separated and purified by HPLC.
Example 77 (Compound 249)
6-(2-(trans-4-(aminomethyl)cyclohexylamino)-5-chloropyridin-4-yl)-N-methyl-
-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
##STR00119##
[0614] Step 1. Preparation of
6-chloro-N-methyl-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
[0615] A mixture of 2,6-dichloropyrazine (298 mg, 2.000 mmol), DMSO
(6 ml), TEA (0.418 ml, 3.00 mmol) and
N-methyl-1-(tetrahydro-2H-pyran-4-yl)methanamine (264 mg, 2.040
mmol) was stirred at 70.degree. C. for 16 hours, and the reaction
progress was followed by LCMS. The crude reaction mixture was let
cool to room temperature, diluted with 150 ml of ethyl acetate,
washed with 1M NaOH soln. (1.times.), water (2.times.), saturated
salt soln. (1.times.), dried with sodium sulfate, filtered, and
concentrated to constant mass, giving 475 mg of the title compound
as free base, which was used without further purification. LCMS
(m/z): 242.0 (MH+), retention time=0.85 min.
Step 2. Preparation of
6-(5-chloro-2-fluoropyridin-4-yl)-N-methyl-N-((tetrahydro-2H-pyran-4-yl)m-
ethyl)pyrazin-2-amine
[0616] To
6-chloro-N-methyl-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2--
amine (450 mg, 1.862 mmol) was added
5-chloro-2-fluoropyridin-4-ylboronic acid (588 mg, 3.35 mmol),
PdCl2(dppf).CH2Cl2 adduct (182 mg, 0.223 mmol), DME (8 ml) and 2M
sodium carbonate (2.79 ml, 5.59 mmol). The resulting reaction
mixture was stirred at 110-115.degree. C. for 90 minutes, and the
reaction progress was followed by LCMS. The reaction mixture was
cooled, 20 ml of ethyl acetate and 10 ml of methanol were added,
filtered and concentrated to crude product. The crude was purified
by silica gel chromatography using 24 g column eluting with 20-75%
ethyl acetate in heptane. The desired fractions were concentrated
to constant mass, giving 499 mg of titled compound as free base.
LCMS (m/z): 337.1 (MH+), retention time=0.90 min.
Step 3. Preparation of
6-(2-(trans-4-(aminomethyl)cyclohexylamino)-5-chloropyridin-4-yl)-N-methy-
l-N-((tetrahydro-2H-pyran-4-yl)methyl)pyrazin-2-amine
[0617] A mixture of
6-(5-chloro-2-fluoropyridin-4-yl)-N-methyl-N-((tetrahydro-2H-pyran-4-yl)m-
ethyl)pyrazin-2-amine (15 mg, 0.045 mmol), DMSO (0.4 ml), and
tert-butyl (trans-4-aminocyclohexyl)methylcarbamate (92 mg, 0.401
mmol) was stirred at 100-105.degree. C. for 18 hours, and the
reaction progress was followed by LCMS. To the crude intermediate
was added 6 M aq. HCl (120 .mu.l, 0.720 mmol) and heated at
80.degree. C. for 40 minutes, and the reaction progress was
followed by LCMS. The reaction mixture was let cool, added 0.5 ml
of DMSO, filtered and purified by prep LC. After lypohilization,
15.6 mg of the title compound, as a TFA salt was obtained. LCMS
(m/z): 445.2 (MH+), retention time=0.59 min.; 1H NMR (300 MHz,
METHANOL-d4, 25.degree. C.) .delta. ppm 1.12-1.47 (m, 6H) 1.59 (d,
J=12.60 Hz, 2H) 1.67 (ddd, J=7.18, 3.81, 3.66 Hz, 1H) 1.92 (d,
J=12.31 Hz, 2H) 2.01-2.11 (m, 1H) 2.16 (d, J=11.43 Hz, 2H) 2.83 (d,
J=7.03 Hz, 2H) 3.17 (s, 3H) 3.33-3.45 (m, 2H) 3.56 (d, J=7.33 Hz,
2H) 3.60-3.72 (m, 1H) 3.93 (dd, J=11.14, 2.93 Hz, 2H) 6.92 (s, 1H)
8.02 (d, J=2.64 Hz, 2H) 8.11 (s, 1H).
Example 78 (Compound 244)
N-(trans-4-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyrazin-2-
-yl)pyridin-2-yl-amino)cyclohexyl)acetamide
##STR00120##
[0618] Step 1: Preparation of
N-(trans-4-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyrazin--
2-yl)pyridin-2-yl-amino)cyclohexyl)acetamide
[0619] A mixture of
trans-N1-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyrazin-2--
yl)pyridin-2-yl)cyclohexane-1,4-diamine (example 85) (14 mg, 0.034
mmol), DCM (0.5 ml), THF (0.500 ml), TEA (0.014 ml, 0.101 mmol) and
acetic anhydride (3.48 .mu.l, 0.037 mmol) was stirred at ambient
temperature for 1 hour, and the reaction progress was followed by
LCMS. The solvent was concentrated off, added 1.0 ml of DMSO,
filtered and purified by prep LC. After lypohilization 6.3 mg of
title compound was obtained as a TFA salt. LCMS (m/z): 445.2 (MH+),
retention time=0.59 min.; 1H NMR (300 MHz. METHANOL-d4, 25.degree.
C.) .delta. ppm 1.21-1.55 (m, 6H) 1.70 (d, J=12.89 Hz, 2H) 1.92 (s,
3H) 1.93-2.06 (m, 3H) 2.10 (br. s., 2H) 3.28-3.32 (dMeOH, 2H App.)
3.34-3.47 (m, 2H) 3.55-3.73 (m, 2H) 3.94 (dd, J=11.28, 3.08 Hz, 2H)
7.00 (s, 1H) 7.94 (s, 2H) 8.01 (s, 1H).
Example 79 (Compound 254)
3,5'-dichloro-N2'-(trans-4-(2-(methylsulfonyl)ethylamino)cyclohexyl)-N6-((-
tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00121##
[0620] Step 1. Preparation of
3,5'-dichloro-N2'-(trans-4-(2-(methylsulfonyl)ethylamino)cyclohexyl)-N6-(-
(tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
[0621] A mixture of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine (Example 87) (40 mg, 0.089
mmol), potassium carbonate (30.7 mg, 0.222 mmol), DMSO (0.4 ml) and
2-(methylsulfonyl)ethyl methanesulfonate (Example 20, step 1) (26.9
mg, 0.133 mmol) was stirred at 100.degree. C. and the reaction
progress was followed by LCMS. After 4 hours, to the crude reaction
mixture was added 2-(methylsulfonyl)ethyl methanesulfonate (26.9
mg, 0.133 mmol) and stirred at 100.degree. C. for an additional 4
hours. The reaction mixture was cooled to room temperature, 0.5 mL
of DMSO added, filtered and purified by prep. LC. After
lypohilization to TFA salt, 16.9 mg of title compound was obtained.
LCMS (m/z): 556.2 (MH+), retention time=0.61 min.; 1H NMR (300 MHz,
METHANOL-d4, 25.degree. C.) .delta. ppm 1.18-1.53 (m, 4H) 1.53-1.72
(m, 4H) 1.86 (dddd, J=14.83, 7.58, 3.96, 3.81 Hz, 1H) 2.24 (d,
J=10.55 Hz, 4H) 3.11 (s, 3H) 3.19 (d, J=6.74 Hz, 2H) 3.25 (br. s.,
1H) 3.38 (td, J=11.72, 1.76 Hz, 2H) 3.56 (s, 4H) 3.72 (t, J=11.28
Hz, 1H) 3.92 (dd, J=11.28, 2.78 Hz, 2H) 6.61 (d, J=9.08 Hz, 1H)
6.67-6.77 (m, 1H) 7.50 (d, J=9.08 Hz, 1H) 8.05 (s, 1H).
Example 80 (Compound 258)
3,5'-dichloro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tetrahydr-
o-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00122##
[0622] Step 1: Preparation of
3,5'-dichloro-N2'-(trans-4-(2-methoxyethylamino)cyclohexyl)-N6-((tetrahyd-
ro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
[0623] A mixture of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine (example 87)(40 mg, 0.089
mmol), potassium carbonate (30.7 mg, 0.222 mmol), DMSO (0.4 ml) and
1-bromo-2-methoxyethane (18.52 mg, 0.133 mmol) was stirred at
80.degree. C. for 2 hours and the reaction progress was followed by
LCMS. To the crude reaction mixture was added BOC-Anhydride (0.041
mL, 0.178 mmol) and stirred at ambient temperature for 2 hr. The
BOC intermediate was purified by prep. LC and lyophilized to TFA
salt, which was then mixed with 4M HCL (1 mL, 4.00 mmol) and
stirred at ambient temperature for 1 hour. The solvent was
concentrated off, the resulting residue dissolved in 1 ml DMSO,
filtered and purified by prep. LC. After lypohilization to TFA
salt, 5.3 mg of the title compound was obtained. LCMS (m/z): 508.2
(MH+), retention time=0.63 min,
Example 81 (Compound 259)
2-(trans-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2,4'-b-
ipyridin-2'-yl-amino)cyclohexylamino)ethanol
##STR00123##
[0624] Step 1. Preparation of
2-(trans-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2,4'--
bipyridin-2'-yl-amino)cyclohexylamino)ethanol
[0625] A mixture of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine (example 87) (40 mg, 0.089
mmol), potassium carbonate (30.7 mg, 0.222 mmol), DMSO (0.4 ml) and
2-bromoethanol (16.65 mg, 0.133 mmol) was stirred at 80.degree. C.
for 2 hours and the reaction progress was followed by LCMS To this
crude reaction mixture was added BOC-Anhydride (0.041 mL, 0.178
mmol) and stirred at ambient temperature for 2 hr. The BOC
intermediate was purified by prep LC, and lyophilized to a TFA
salt. Then was added 4M HCl in Dioxane (1 mL. 4.00 mmol) and
stirred at ambient temperature for 1 hour. The solvent was
concentrated off, the resulting residue dissolved in DMSO, purified
by prep. LC. After lypohilization to TFA salt, 6.1 mg of the title
compound was obtained. LCMS (m/z): 494.2 (MH+), retention time=0.60
min.; 1H NMR (300 MHz, METHANOL-d4, 25.degree. C.) .delta. ppm
1.18-1.51 (m, 4H) 1.50-1.73 (m, 4H) 1.78-1.95 (m, J=14.80, 7.62,
7.47, 3.66, 3.66 Hz, 1H) 2.23 (d, J=11.43 Hz, 4H) 3.09-3.24 (m, 5H)
3.38 (td, J=11.79, 1.61 Hz, 2H) 3.64-3.77 (m, 1H) 3.77-3.84 (m, 2H)
3.92 (dd. J=11.28, 3.08 Hz, 2H) 6.59 (d, J=9.08 Hz, 1H) 6.66 (s,
1H) 7.49 (d, J=8.79 Hz, 1H) 8.03 (s, 1H)
Example 82 (Compound 265)
N2'-(trans-4-aminocyclohexyl)-3-chloro-N6-((tetrahydro-2H-pyran-4-yl)methy-
l)-2,4'-bipyridine-2',6-diamine
##STR00124##
[0626] Step 1. Preparation of
3-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridin-6--
amine
[0627] A mixture of
6-bromo-5-chloro-N-((tetrahydro-2H-pyran-4-yl)methyl)pyridin-2-amine
(intermediate E) (630 mg, 2.062 mmol), 2-fluoropyridin-4-ylboronic
acid (639 mg, 4.54 mmol), PdCl.sub.2(dppf).CH2Cl2 adduct (168 mg,
0.206 mmol), DME (9 ml) and 2M sodium carbonate (3.09 ml, 6.18
mmol) was stirred at 105.degree. C. for 2 hours, and the reaction
progress was followed by LCMS. The reaction mixture was let cool to
room temperature, diluted with 30 ml of ethyl acetate, 10 ml of
methanol, filtered and concentrated. The crude material was
purified by silica gel chromatography using 40 g column and eluting
with 5-45% ethyl acetate in heptane. The desired fractions were
concentrated to constant mass giving, 516 mg of the title compound
as free base. LCMS (m/z): 332.0 (MH+), retention time=0.88 min
Step 2. Preparation of
N2'-(trans-4-aminocyclohexyl)-3-chloro-N6-((tetrahydro-2H-pyran-4-yl)meth-
yl)-2,4'-bipyridine-2',6-diamine
[0628] A mixture of
3-chloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridin-6--
amine (250 mg, 0.777 mmol), DMSO (2 ml), and
trans-cyclohexane-1,4-diamine (798 mg, 6.99 mmol) was stirred at
105.degree. C. for 20 hours and the reaction progress was followed
by LCMS. The crude reaction mixture was cooled to room temperature,
diluted with 250 ml of ethyl acetate, washed with saturated sodium
bicarbonate (1.times.), water (2.times.), filtered and the solvent
was concentrated off. The crude was dissolved in 5 ml DMSO,
filtered and purified by prep. LC. After lyapholization to TFA
salt, 180 mg of the title compound was obtained. LCMS (m/z): 416.2
(MH+), retention time=0.52 min.; 1H NMR (300 MHz, METHANOL-d4,
25.degree. C.) .delta. ppm 1.20-1.41 (m, 2H) 1.46-1.74 (m, 6H) 1.85
(ddd, J=10.99, 7.33, 4.25 Hz, 1H) 2.06-2.30 (m, 4H) 3.19 (br. s.,
1H) 3.26 (d, J=7.03 Hz, 2H) 3.33-3.46 (m, 2H) 3.59-3.76 (m, 1H)
3.93 (dd, J=11.14, 3.22 Hz, 2H) 6.60 (d, J=8.79 Hz, 1H) 7.23 (d,
J=6.74 Hz, 1H) 7.39 (s, 1H) 7.49 (d, J=8.79 Hz, 1H) 7.88 (d, J=6.74
Hz, 1H)
Example 83 (Compound 268)
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((R)-tet-
rahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine
##STR00125##
[0629] Step 1. Preparation of (R)-(tetrahydrofuran-2-yl)methyl
methanesulfonate
[0630] A mixture of (R)-(tetrahydrofuran-2-yl)methanol (600 mg,
5.87 mmol), DCM (35 ml), TEA (0.983 ml, 7.05 mmol) was diluted with
methanesulfonyl chloride (0.467 ml, 5.99 mmol), via a dropwise
addition. The reaction mixture was stirred at ambient temperature
for 5 hours and the reaction progress was followed by LCMS. The
crude reaction mixture was washed with saturated sodium bicarbonate
(1.times.), water (2.times.), filtered and concentrated to a
constant mass, giving 980 mg of the title compound, which was used
without further purification. LCMS (m/z): 181.0 (MH+), retention
time=0.40 min
Step 2. Preparation of
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(((R)-te-
trahydrofuran-2-yl)methyl)aminocyclohexyl)-2,4'-bipyridine-2',6-diamine
[0631] To
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-p-
yran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine (example 87) (40 mg,
0.089 mmol) was added potassium carbonate (30.7 mg, 0.222 mmol),
DMSO (0.4 ml) and (R)-(tetrahydrofuran-2-yl)methyl methanesulfonate
(24.01 mg, 0.133 mmol), and the resulting reaction mixture was
stirred at 100.degree. C. for 4 hours and the reaction progress was
followed by LCMS. After about 4 hours
(R)-(tetrahydrofuran-2-yl)methyl methanesulfonate (24.01 mg, 0.133
mmol) was added and the resulting mixture was stirred at
100.degree. C. for 4 hours more. The reaction mixture was cooled to
room temperature, 0.5 mL of DMSO added, filtered and purified by
prep. LC. After lyapholization to a TFA salt, 9.1 mg of the title
compound was obtained. LCMS (m/z): 534.3 (MH+), retention time=0.62
min.
Example 84 (Compound 272)
3,5'-dichloro-N2'-(trans-4-((2-methoxyethyl)(methyl)amino)cyclohexyl)-N6-(-
(tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
##STR00126##
[0632] Step 1. Preparation of
(1s,4s)-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2,4'-b-
ipyridin-2'-yl-amino)cyclohexanol
[0633] To
3,5'-dichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4-
'-bipyridin-6-amine (intermediate G) (712 mg, 1.999 mmol) was added
DMSO (4.5 ml), TEA (1.114 ml, 8.00 mmol) and
(1s,4s)-4-aminocyclohexanol (607 mg, 4 mmol), and the reaction
mixture was stirred at 95-100.degree. C. for 96 hours and the
reaction progress was followed by LCMS. The reaction mixture was
cooled, 250 ml of ethyl acetate was added, washed with saturated
sodium bicarbonate (1.times.) water (2.times.) and concentrated to
constant mass. The crude was purified by silica gel chromatography
using 40 g column eluting with 25-95% ethyl acetate in heptane. The
desired fractions were concentrated to constant mass, giving 380 mg
of title compound as free base. LCMS (m/z): 451.1 (MH+), retention
time=0.65 min
Step 2. Preparation of
(1s,4s)-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)amino-2,4'-b-
ipyridin-2'-yl-amino)cyclohexyl methanesulfonate
[0634] To
(1s,4s)-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)ami-
no-2,4'-bipyridin-2'-yl-amino)cyclohexanol (375 mg, 0.831 mmol) was
added DCM (8 ml), and TEA (0.174 ml, 1.246 mmol) and the resulting
mixture was cooled in an ice bath to 0.degree. C. Then with
stirring was added methanesulfonyl chloride (0.071 ml, 0.914 mmol).
The reaction mixture was allowed to warm to ambient temperature and
stirred for 2 hours, and the reaction progress was followed by
LCMS. To the crude reaction mixture was added 250 ml of ethyl
acetate, washed with saturated sodium bicarbonate (1.times.) water
(2.times.) and concentrated to constant mass giving, 441 mg of
title compound as free base, used without further purification.
LCMS (m/z): 529.3 (MH+), retention time=0.75 min.
Step 3. Preparation of
3,5'-dichloro-N2'-(trans-4-((2-methoxyethyl)(methyl)amino)cyclohexyl)-N6--
((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine
[0635] To
(1s,4s)-4-(3,5'-dichloro-6-((tetrahydro-2H-pyran-4-yl)methyl)ami-
no-2,4'-bipyridin-2'-yl-amino)cyclohexyl methanesulfonate (48 mg,
0.091 mmol) was added t-Butanol (0.22 ml) and
2-methoxy-N-methylethanamine (202 mg, 2.266 mmol). The reaction
mixture was stirred at 95-100.degree. C. for 5 hours and the
reaction progress was followed by LCMS. The reaction mixture was
cooled to room temperature, 12 ml of ethyl acetate was added then
washed with saturated sodium bicarbonate (1.times.) water
(2.times.) and the solvent concentrated off. The resulting residue
was dissolved in 1 ml of DMSO, filtered and purified by prep LC.
After lyapholization to TFA salt, 8.61 mg of the title compound was
obtained. LCMS (m/z): 522.2 (MH+), retention time=0.63 min.; 1H NMR
(300 MHz, METHANOL-d4, 25.degree. C.) .delta. ppm 1.18-1.56 (m, 4H)
1.59-1.79 (m, 4H) 1.79-1.95 (m, 1H) 2.02-2.35 (m, 4H) 2.87 (s, 3H)
3.19 (d, J=6.74 Hz, 2H) 3.24 (d, J=3.52 Hz, 1H) 3.32-3.41 (m, 3H)
3.42 (s, 3H) 3.46-3.58 (m, 1H) 3.63-3.78 (m, 3H) 3.92 (dd, J=11.14,
2.93 Hz, 2H) 6.60 (d, J=9.08 Hz, 1H) 6.70 (s, 1H) 7.49 (d, J=9.08
Hz, 1H) 8.04 (s, 1H)
Example 85 (Compound 203)
trans-N1-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyrazin-2-y-
l)pyridin-2-yl)cyclohexane-1,4-diamine
##STR00127##
[0636] Step 1. Preparation of
trans-N1-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyrazin-2--
yl)pyridin-2-yl)cyclohexane-1,4-diamine
[0637] To
6-(5-chloro-2-fluoropyridin-4-yl)-N-((tetrahydro-2H-pyran-4-yl)m-
ethyl)pyrazin-2-amine (example 75 step 2) (20 mg, 0.062 mmol) was
added DMSO (0.6 ml) and trans-cyclohexane-1,4-diamine (63.7 mg,
0.558 mmol). The reaction mixture then was stirred at
100-105.degree. C. for 18 hours and the reaction progress was
followed by LCMS. The reaction mixture was let cool, diluted with
0.5 ml of DMSO, filtered and purified by prep LC. After
lyapholization to TFA salt, 13.7 mg of the title compound was
obtained. LCMS (m/z): 417.3 (MH+), retention time=0.46 min.; 1H NMR
(300 MHz, METHANOL-d4, 25.degree. C.) .delta. ppm 1.22-1.78 (m, 8H)
1.81-2.01 (m, 1H) 2.03-2.28 (m, 4H) 3.05-3.21 (m, 1H) 3.28-3.32
(dMeOH, 2H App.) 3.39 (td, J=11.72, 1.76 Hz, 2H) 3.62-3.79 (m, 1H)
3.94 (dd, J=11.14, 3.22 Hz, 2H) 6.95 (s, 1H) 7.92 (d, J=2.93 Hz,
2H) 8.05 (s, 1H).
Example 86 (Compound 243)
trans-N1-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyrazin-2-y-
l)pyridin-2-yl)-N4-(2-methoxyethyl)cyclohexane-1,4-diamine
##STR00128##
[0638] Step 1. Preparation of
trans-N1-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminopyrazin-2--
yl)pyridin-2-yl)-N4-(2-methoxyethyl)cyclohexane-1,4-diamine
[0639] To
trans-N1-(5-chloro-4-(6-((tetrahydro-2H-pyran-4-yl)methyl)aminop-
yrazin-2-yl)pyridin-2-yl)cyclohexane-1,4-diamine (Example 85) (16
mg, 0.038 mmol) was added DMSO (0.4 ml), potassium carbonate (15.91
mg, 0.115 mmol) and 1-bromo-2-methoxyethane (7.47 mg, 0.054 mmol).
The reaction mixture then was stirred at 70.degree. C. for 6 hours
and the reaction progress was followed by LCMS. The reaction
mixture was cooled to room temperature, 0.5 ml of DMSO was added,
filtered and purified by prep LC. After lyapholization to TFA salt,
2.7 mg of the title compound was obtained. LCMS (m/z): 475.2 (MH+),
retention time=0.51 min.
Example 87 (Compound 253)
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)-
methyl)-2,4'-bipyridine-2',6-diamine
##STR00129##
[0640] Step 1. Preparation of
N2'-(trans-4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl-
)methyl)-2,4'-bipyridine-2',6-diamine
[0641] To
3,5'-dichloro-2'-fluoro-N-((tetrahydro-2H-pyran-4-yl)methyl)-2,4-
'-bipyridin-6-amine (Intermediate G) (250 mg, 0.702 mmol) was added
DMSO (3 ml) and trans-cyclohexane-1,4-diamine (952 mg, 6.32 mmol).
The reaction mixture was stirred at 100.degree. C. for 20 hours and
the reaction progress was followed by LCMS. The reaction mixture
was cooled, diluted with 250 ml ethyl acetate, washed with
saturated sodium bicarbonate (1.times.), water (3.times.) and
concentrated to constant mass, giving 320 mg of product as a free
base, which was used without further purification. A portion of the
title compound, 25 mg was purified by prep LC and lyapholized to
give 17.6 mg of the title compound as a TFA salt. LCMS (m/z): 450.2
(MH+), retention time=0.58 min.; 1H NMR (300 MHz, METHANOL-d4,
25.degree. C.) .delta. ppm 1.16-1.76 (m, 8H) 1.76-1.98 (m, 1H)
2.04-2.27 (m, 4H) 3.06-3.16 (m, 1H) 3.19 (d, J=6.74 Hz, 2H) 3.37
(t, J=11.87 Hz, 2H) 3.62-3.77 (m, 1H) 3.92 (dd, J=11.28, 3.08 Hz,
2H) 6.61 (d, J=8.79 Hz, 1H) 6.73 (s, 1H) 7.50 (d, J=9.08 Hz, 1H)
8.04 (s, 1H).
Example 88 (Compound 178)
5'-chloro-N6-(3-fluorobenzyl)-N2'-methyl-2,4'-bipyridine-2',6-diamine
##STR00130##
[0642] Step 1. Preparation of
5'-chloro-N6-(3-fluorobenzyl)-N2'-methyl-2,4'-bipyridine-2',6-diamine
[0643] A mixture of
5'-chloro-2'-fluoro-N-(3-fluorobenzyl)-2,4'-bipyridin-6-amine
(Intermediate B) (15 mg, 0.045 mmol) was added DMSO (0.4 ml) and
methyl amine 40% in water (200 .mu.l, 2.293 mmol) in a microwave
tube was microwaved at 145.degree. C. for 900 seconds and the
reaction progress was followed by LCMS. Most of the amine was
removed under vacuum, 0.5 ml of DMSO was added, filtered and
purified by prep LC. After lyapholization 13.9 mg of the title
compound was obtained as a TFA salt. LCMS (m/z): 343.0 (MH+),
retention time=0.67 min.; 1H NMR (300 MHz, METHANOL-d4, 25.degree.
C.) .delta. ppm 2.97 (s, 3H) 4.62 (s, 2H) 6.81 (d, J=8.50 Hz, 1H)
6.91-7.02 (m, 3H) 7.09 (d, J=9.96 Hz, 1H) 7.17 (d, J=7.62 Hz, 1H)
7.27-7.39 (m, 1H) 7.69 (dd, J=8.50, 7.33 Hz, 1H) 8.03 (s, 1H).
Example 89 (Compound 332)
5'-chloro-5-fluoro-N6-((4-methyltetrahydro-2H-pyran-4-yl)methyl)-N2'-(tran-
s-4-((1,1-dioxo-hexahydro-1hexahydro-1-thiopyran-4-yl)-3-ylamino)cyclohexy-
l)-2,4'-bipyridene-2', 6-diamine
##STR00131##
[0645] To a solution of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-((4-methyltetrahydro--
2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine (40 mg, 0.089
mmol) in DMF (0.5 ml) was added potassium carbonate (49.4 mg, 0.357
mmol), 3-chloro-1,1-dioxo-tetrahydro-1-thiophene (83 mg, 0.536
mmol) and sodium iodide (40.2 mg, 0.268 mmol). The reaction mixture
was stirred at 100.degree. C. for 42 hours. The cooled reaction
mixture was diluted with water and extracted with ethyl acetate.
The combined extracts were washed sequentially with water and
brine, dried over sodium sulfate, filtered, and concentrated. The
residue was purified by reverse phase HPLC and lyophilized to give
3.8 mg off-white powder of the title compound as its TFA salt. LCMS
(m/z): 566.2 (MH+), retention time=0.64 min.
Example 90 (Compound 333)
5'-chloro-5-fluoro-N2'-(trans-4-((2-methyl-1,3-dioxolan-2-yl)methyl)aminoc-
yclohexyl)-N6-((tetrahydro-2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diam-
ine
##STR00132##
[0647] To a solution of
N2'-(trans-4-aminocyclohexyl)-5'-chloro-5-fluoro-N6-((4-methyltetrahydro--
2H-pyran-4-yl)methyl)-2,4'-bipyridine-2',6-diamine (21 mg, 0.048
mmol) in DCM (1.0 ml) was added
2-methyl-1,3-dioxolane-2-carbaldehyde (synthesized following the
procedure reported in Org. Lett., 2009, 11, 3542-3545), sodium
triacetoxyborohydride (20.51 mg, 0.097 mmol). The reaction mixture
was stirred at ambient temperature for 2 hours. The reaction
mixture was diluted with water and extracted with ethyl acetate.
The combined extracts were washed sequentially with water and
brine, dried over sodium sulfate, filtered, and concentrated. The
residue was purified by reverse phase HPLC and lyophilized to give
12 mg off-white powder of the title compound as its TFA salt. LCMS
(m/z): 534.1 (MH+), retention time=0.62 min.
Example 91 (Compound 349)
(4-((5'-chloro-2'-(trans-4-((R)-1-methoxypropan-2-ylamino)cyclohexylamino)-
-2,4'-bipyridin-6-ylamino)methyl)tetrahydro-2H-pyran-4-yl)methanol
##STR00133##
[0648] Step 1. Synthesis of methyl
4-cyanotetrahydro-2H-pyran-4-carboxylate
[0649] To 1-bromo-2-(2-bromoethoxy)ethane (2.57 g, 11.10 mmol) in
DMSO (6 mL, by mistake, should use DMF) at room temperature was
added methylcyanoacetate (1 g, 10.09 mmol) and DBU (3.35 ml, 22.20
mmol) sequentially. The brown mixture was heated to 85.degree. C.
in a capped glass vial for 3 hours. The resulting solution was dark
brown.
[0650] The reaction mixture was poured into water and extracted
with EtOAc. The organic extracts were combined, washed with water,
brine, dried with sodium sulfate and concentrated in vacuo to give
0.944 g of brown oil. This crude material was used in the next step
without further purification.
Step 2. Synthesis of
(4-(aminomethyl)tetrahydro-2H-pyran-4-yl)methanol
[0651] To the crude product from step 1 (0.944 g, 5.58 mmol) in THF
(5 ml) (a dark brown solution) at 0.degree. C. was added LAH (5.58
ml, 5.58 mmol) dropwise via a syringe. The brown mixture was warmed
to room temperature and stirred for 18 hours. The resulting mixture
was yellow cloudy. LC/MS showed containing desired product. To the
reaction was added sodium sulfate decahydrate solid at 0.degree. C.
The mixture was stirred at room temperature for 20 min., then
filtered and washed with DCM. The yellow filtrate was concentrated
in vacuo to give 0.74 g of orange oil. This crude material was used
in the next step without further purification.
Step 3. Synthesis of
(4-((6-bromopyridin-2-ylamino)methyl)tetrahydro-2H-pyran-4-yl)methanol
[0652] To 2-bromo-6-fluoropyridine (0.448 g, 2.55 mmol) in NMP (4
ml) at room temperature was added TRIETHYLAMINE (0.852 ml, 6.12
mmol) and the crude product obtained in step 2 (370 mg, 2.55 mmol)
sequentially. The yellow mixture was heated to 75.degree. C. in a
capped glass vial for 3 hours. LC/MS showed about 20% conversion to
the product. Continued heating at 110.degree. C. for 16 hrs. The
reaction mixture was cooled to room temperature, poured into water
and extracted with EtOAc. The organic extracts were combined,
washed with water, brine, dried with sodium sulfate and
concentrated in vacuo to give 0.5 g of brown oil. The crude mixture
was purified by Analogix system (silica gel column 24 g, gradient:
0 min, 100% n-hexane; 2-127 min, 10% EtOAc in Hex; 7-13 min. 20%
EtOAc in Hex; 13-16 min. 30% EtOAc in Hex; 16-30 min. 50% EtOAc in
Hex; 30-35 min. 100% EtOAc). The pure fractions were combined and
concentrated in vacuo to give 0.13 g of desired product as a white
crystal. LCMS (m/z): 301/303 (MH+), retention time=0.67 min.
Step 4. Synthesis of
(4-((5'-chloro-2'-fluoro-2,4'-bipyridin-6-ylamino)methyl)tetrahydro-2H-py-
ran-4-yl)methanol
[0653] Following the same procedure as in Example 1b using
(4-((6-bromopyridin-2-ylamino)methyl)tetrahydro-2H-pyran-4-yl)methanol
(from step 3) and 5-chloro-2-fluoropyridin-4-ylboronic acid, the
desired product was obtained. LCMS (m/z): 352 (MH+), retention
time=0.54 min.
Step 5. Synthesis of
(4-((5'-chloro-2'-(trans-4-((R)-1-methoxypropan-2-ylamino)cyclohexylamino-
)-2,4'-bipyridin-6-ylamino)methyl)tetrahydro-2H-pyran-4-yl)methanol
[0654] Following the same procedure as in Example 1b using
(4-((5'-chloro-2'-fluoro-2,4'-bipyridin-6-ylamino)methyl)tetrahydro-2H-py-
ran-4-yl)methanol (from step 4) and
trans-N1-((R)-1-methoxypropan-2-yl)cyclohexane-1,4-diamine, the
desired product was obtained. LCMS (m/z): 518.2 (MH+), retention
time=0.47 min.
Example 92 (Compound 348)
3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methyl)-N2'-(trans-4-(1,1-diox-
o-tetrahydro-2H-thiopyran-4-ylamino)cyclohexyl)-2,4'-bipyridine-2',6-diami-
ne
##STR00134##
[0656] Compound
N2'-(4-aminocyclohexyl)-3,5'-dichloro-N6-((tetrahydro-2H-pyran-4-yl)methy-
l)-2,4'-bipyridine-2',6-diamine (0.100 g, 0.222 mmol) (synthesized
in the same manner as in Example 1b),
2,3,5,6-tetrahydro-4H-thiopyran-4-one 1,1-dioxide (0.036 g, 0.244
mmol), and triethylamine (0.251 ml, 0.182 g, 1.798 mmol) were
dissolved in anhydrous CH.sub.2Cl.sub.2 (1.0 ml) and placed under
argon. This solution was then treated with sodium
triacetoxyborohydride (0.094 g, 0.444 mmol). The reaction was then
stirred at room temperature for 18 hours. At this time a LC-MS was
run. The reaction was about 25% complete. Additional
2,3,5,6-tetrahydro-4H-thiopyran-4-one 1,1-dioxide (.about.4
equivalents) and sodium triacetoxy borohydride (.about.8
equivalents) were added and the reaction continued for additional
27 hours. The reaction was about 60% complete as indicated by
LC/MS. The reaction was quenched with sat NaHCO.sub.3 (15 ml). This
was extracted with EtOAc (3.times.15 ml). The combined extracts
were washed with brine (1.times.15 ml), dried (Na.sub.2SO.sub.4),
filtered and the solvent removed in vacuo. The material was
purified using the HPLC and lyophilized to give 19.7 mg off-white
powder of the title compound as its TFA salt. LCMS (m/z): 582/584
(MH+), retention time=0.58 min.
Example 93 (Compound 310)
4-((5'-chloro-2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cyclohexylamino)-
-2,4'-bipyridin-6-ylamino)methyl)tetrahydro-2H-pyran-4-carbonitrile
##STR00135##
[0658] This compound was synthesized following the procedure of
Example 1b using Intermediates AB (40 mg, 0.115 mmol) and
N1-((R)-1-methoxypropan-2-yl)cyclohexane-trans-1,4-diamine
(synthesized in step 2 of Example 67, 107 mg, 0.577 mmol). The
product was obtained as an off white powder (30.2 mg, 35.5% yield).
LCMS (m/z): 513.2 [M+H]+; retention time=0.531 min. 1H NMR (400
MHz, CHLOROFORM-d) .delta. ppm 1.04 (d, J=6.26 Hz, 2H) 1.12-1.37
(m, 3H) 1.84-2.06 (m, 3H) 2.10-2.25 (m, 2H) 2.44-2.69 (m, 1H)
2.91-3.11 (m, 1H) 3.20-3.39 (m, 3H) 3.43-3.60 (m, 1H) 3.61-3.83 (m,
3H) 3.90-4.08 (m, 2H) 4.41 (d, J=8.22 Hz, 1H) 4.67-4.93 (m, 1H)
6.37-6.62 (m, 2H) 6.97 (d, J=7.43 Hz, 1H) 7.26 (s, 1H) 7.39-7.58
(m, 1H).
Example 94 (Synthesis of Compound 340)
Synthesis of
4-((5'-chloro-5-fluoro-2'-(trans-4-((R)-1-methoxypropan-2-yl-amino)cycloh-
exylamino)-2,4'-bipyridin-6-ylamino)methyl)tetrahydro-2H-pyran-4-carbonitr-
ile
##STR00136##
[0660] This compound was synthesized following the procedure of
Example 1a using intermediates AA (50 mg, 0.137 mmol) and
N1-((R)-1-methoxypropan-2-yl)cyclohexane-trans-1,4-diamine
(synthesized in step 2 of Example 67, 128 mg, 0.685 mmol). The
product was obtained as an off white powder 35 mg (33.6% yield).
LCMS (m/z): 531.2 [M+H]+; retention time=0.595 min.
[0661] Examples in Table 1 were prepared using methods analogous to
those described above. The method column in Table 1 indicates the
synthetic procedure, from a specific example, used to synthesize a
given compound. Thus for example, Compound 7 is synthesized by the
procedure outlined in Example 7, while compound 25 is synthesized
by the procedure outlined in Example 1a, and the like.
TABLE-US-00001 TABLE 1 retention Com- M + H time pound # Structure
(m/z) (min.) method 1 ##STR00137## 426.2 0.7 Example 1a 2
##STR00138## 380.3 0.61 Example 2 3 ##STR00139## 415.3 0.67 Example
3 4 ##STR00140## 440.2 0.62 Example 4 5 ##STR00141## 412.2 0.6
Example 5 6 ##STR00142## 504.2 0.77 Example 6 7 ##STR00143## 511.3
0.62 Example 7 8 ##STR00144## 402.3 0.41 Example 8 9 ##STR00145##
441.3 0.76 Example 9 10 ##STR00146## 483.2 0.65 Example 10 11
##STR00147## 460.3 0.72 Example 11 12 ##STR00148## 444.2 0.7
Example 12 13 ##STR00149## 451.2 0.67 Example 13 14 ##STR00150##
469.1 0.56 Example 14 15 ##STR00151## 451.2 0.7 Example 15 16
##STR00152## 469.2 0.56 Example 16 17 ##STR00153## 470.2 0.61
Example 17 18 ##STR00154## 454.2 0.61 Example 18 19 ##STR00155##
470.3 0.58 Example 19 20 ##STR00156## 532.2 0.62 Example 20 21
##STR00157## 440.3 0.61 Example 21 22 ##STR00158## 454.2 0.64
Example 22 23 ##STR00159## 498.3 0.65 Example 23 24 ##STR00160##
414.3 0.72 Example 1a 25 ##STR00161## 408.2 0.61 Example 1a 26
##STR00162## 409.2 0.41 Example 1a 27 ##STR00163## 444.2 0.63
Example 1a 27 ##STR00164## 444.2 0.63 Example 1a 29 ##STR00165##
444.3 0.64 Example 1a 30 ##STR00166## 393.2 0.54 Example 10 31
##STR00167## 374.3 0.56 Example 2 32 ##STR00168## 392.3 0.59
Example 2 33 ##STR00169## 375.3 0.36 Example 2 34 ##STR00170##
427.3 0.61 Example 3 35 ##STR00171## 410.3 0.41 Example 3 36
##STR00172## 474.2 0.66 Example 1a 37 ##STR00173## 442.2 0.66
Example 1a 38 ##STR00174## 427.2 0.49 Example 1a 39 ##STR00175##
444.2 0.64 Example 1a 40 ##STR00176## 416.3 0.46 Example 1a 41
##STR00177## 426.3 0.61 Example 1a 42 ##STR00178## 442.2 0.65
Example 1a 43 ##STR00179## 422.3 0.63 Example 1a 44 ##STR00180##
438.3 0.59 Example 1a 45 ##STR00181## 492.2 0.72 Example 1a 46
##STR00182## 487.1/ 489.2 0.53 Example 1a 47 ##STR00183## 476.3
0.69 Example 1a 48 ##STR00184## 486.2/ 488.2 0.67 Example 1a 49
##STR00185## 424.2 0.5 Example 1a 50 ##STR00186## 426.2 0.6 Example
1a 51 ##STR00187## 422.2 0.63 Example 1a 52 ##STR00188## 476.3 0.7
Example 1a 53 ##STR00189## 474.2 0.64 Example 1a 54 ##STR00190##
492.2 0.73 Example 1a 55 ##STR00191## 442.2 0.66 Example 1a 56
##STR00192## 428.3 0.66 Example 4 57 ##STR00193## 422.3 0.6 Example
4 58 ##STR00194## 423.3 0.41 Example 4 59 ##STR00195## 441.3 0.5
Example 4 60 ##STR00196## 423.3 0.41 Example 4 61 ##STR00197##
458.3 0.65 Example 4 62 ##STR00198## 456.3 0.67 Example 4 63
##STR00199## 458.3 0.65 Example 4 64 ##STR00200## 488.3 0.67
Example 4 65 ##STR00201## 458.3 0.66 Example 4 66 ##STR00202##
440.3 0.62 Example 4 67 ##STR00203## 512.3 0.91 Example 5 68
##STR00204## 400.3 0.64 Example 5 69 ##STR00205## 394.3 0.58
Example 5 70 ##STR00206## 413.2 0.47 Example 5 71 ##STR00207##
430.2 0.63 Example 5 72 ##STR00208## 395.2 0.39 Example 5 73
##STR00209## 428.2 0.65 Example 5 74 ##STR00210## 430.2 0.62
Example 5 75 ##STR00211## 428.4 0.68 Example 1a 76 ##STR00212##
422.3 0.7 Example 1a 77 ##STR00213## 440.2 0.73 Example 1a 78
##STR00214## 518.3 0.74 Example 6 79 ##STR00215## 532.3 0.77
Example 6 80 ##STR00216## 537.3 0.63 Example 7 81 ##STR00217##
531.3 0.64 Example 7 82 ##STR00218## 525.3 0.64 Example 7 83
##STR00219## 551.3 0.65 Example 7 84 ##STR00220## 545.3 0.66
Example 7 85 ##STR00221## 497.3 0.62 Example 7 86 ##STR00222##
523.3 0.63 Example 7 87 ##STR00223## 517.3 0.64 Example 7 88
##STR00224## 454.3 0.62 Examples 1, 10 89 ##STR00225## 400.3 0.65
Example 5 90 ##STR00226## 402.2 0.47 Example 5 91 ##STR00227##
412.2 0.61 Example 5 92 ##STR00228## 430.2 0.64 Example 5 93
##STR00229## 395.2 0.41 Example 5 94 ##STR00230## 430.2 0.63
Example 5 95 ##STR00231## 400.3 0.65 Example 5 96 ##STR00232##
402.2 0.47 Example 5 97 ##STR00233## 412.2 0.61 Example 5 98
##STR00234## 430.2 0.64 Example 5 99 ##STR00235## 395.2 0.41
Example 5 100 ##STR00236## 430.2 0.64 Example 5 101 ##STR00237##
427.2 0.6 Example 10 102 ##STR00238## 468.3 0.67 Examples 5, 10 103
##STR00239## 426.2 0.61 Examples 5, 10 104 ##STR00240## 402.3 0.46
Example 1a 105 ##STR00241## 437.3 0.53 Example 1a 106 ##STR00242##
402.3 0.46 Example 1a 107 ##STR00243## 416.3 0.47 Example 1a 105
##STR00244## 462.3 0.78 Example 1a 106 ##STR00245## 418.3 0.43
Example 1a 107 ##STR00246## 427.3 0.75 Example 9 108 ##STR00247##
430.3 0.54 Example 1a 109 ##STR00248## 425.3 0.39 Example 1a 110
##STR00249## 508.2 0.68 Example 1b, intermediate B 111 ##STR00250##
523.3 0.6 Example 1b, intermediate B 112 ##STR00251## 494.2 0.67
Example 1b, intermediate B 113 ##STR00252## 468.2 0.67 Example 1b,
intermediate B 114 ##STR00253## 427.2 0.66 Example 1b, intermediate
B 115 ##STR00254## 494.2 0.64 Example 1b, intermediate B 116
##STR00255## 496.2 0.62 Example 1b, intermediate B 117 ##STR00256##
362.3 0.38 Example 8 118 ##STR00257## 376.3 0.37 Example 8 119
##STR00258## 416.3 0.39 Example 8
120 ##STR00259## 410.2 0.42 Example 8 121 ##STR00260## 416.3 0.43
Example 8 122 ##STR00261## 410.2 0.42 Example 8 123 ##STR00262##
416.3 0.41 Example 8 124 ##STR00263## 404.2 0.39 Example 8 125
##STR00264## 403.1 0.49 Example 8 126 ##STR00265## 417.1 0.48
Example 8 127 ##STR00266## 403.1 0.49 Example 8 128 ##STR00267##
417.1 0.5 Example 8 129 ##STR00268## 403.3 0.47 Example 8 130
##STR00269## 403.3 0.49 Example 8 131 ##STR00270## 389.2 0.58
Example 8 132 ##STR00271## 415.2 0.7 Example 8 133 ##STR00272##
445.2 0.71 Example 8 134 ##STR00273## 427.2 0.66 Example 8 135
##STR00274## 419.3 0.47 Example 8 136 ##STR00275## 416.3 0.45
Example 8 137 ##STR00276## 451.2 0.67 Example 14 138 ##STR00277##
469.1 0.56 Example 15 139 ##STR00278## 426.3 0.61 Example 1b 140
##STR00279## 397.3 0.81 Example 1b 141 ##STR00280## 411.3 0.86
Example 1b 142 ##STR00281## 413.2 0.71 Example 1b 143 ##STR00282##
427.2 0.72 Example 1b 144 ##STR00283## 386.2 0.57 Example 1b 145
##STR00284## 400.3 0.58 Example 1b 146 ##STR00285## 387.2 0.62
Example 1b 147 ##STR00286## 401.2 0.71 Example 1b 148 ##STR00287##
413.2 0.7 Example 1b 149 ##STR00288## 447.3 0.87 Example 1b 150
##STR00289## 413.2 0.69 Example 1b 151 ##STR00290## 481.2 0.63
Example 1b 152 ##STR00291## 404.1 0.62 Example 32 153 ##STR00292##
426.1 0.67 Example 1b 154 ##STR00293## 471 0.75 Example 1b 155
##STR00294## 565.2 0.85 Example 26 156 ##STR00295## 538.1 0.82
Example 27 157 ##STR00296## 484.2 0.63 Example 19 158 ##STR00297##
509.2 0.58 Example 1b 159 ##STR00298## 372.2 0.7 Example 1b 160
##STR00299## 373.2 0.75 Example 1b 161 ##STR00300## 428.3 0.73
Example 1b 162 ##STR00301## 426.3 0.73 Example 1b, Example 8 163
##STR00302## 440.3 0.73 Example 1b 164 ##STR00303## 426.3 0.74
Example 1b, Example 8 165 ##STR00304## 440.3 0.77 Example 1b,
Example 8 166 ##STR00305## 426.3 0.76 Example 1b, Example 8 167
##STR00306## 440.3 0.77 Example 1b, Example 8 168 ##STR00307##
441.3 0.72 Example 1b, Example 8 169 ##STR00308## 440.3 0.76
Example 1b, Example 8 170 ##STR00309## 455.3 0.71 Example 1b 171
##STR00310## 442.2 0.75 Example 1b 172 ##STR00311## 371.2 0.85
Example 1b 173 ##STR00312## 385.2 0.91 Example 1b 174 ##STR00313##
441.3 0.86 Example 1b 175 ##STR00314## 434.2 0.74 Example 1b 176
##STR00315## 427.2 0.85 Example 1b 177 ##STR00316## 434.2 0.73
Example 1b 178 ##STR00317## 343 0.67 Example 88 179 ##STR00318##
329 0.65 Example 88 180 ##STR00319## 422.3 0.54 Example 69 181
##STR00320## 441.2 0.71 Example 1b 182 ##STR00321## 427.1 0.69
Example 1b 183 ##STR00322## 413.2 0.68 Example 1b 184 ##STR00323##
413.2 0.68 Example 1b 185 ##STR00324## 409.3 0.83 Example 21 186
##STR00325## 401.2 0.65 Example 1b 187 ##STR00326## 443.2 0.87
Example 1b 188 ##STR00327## 418.2 0.61 Example 1b 189 ##STR00328##
405.2 0.68 Example 1b 190 ##STR00329## 455.3 0.8 Example 1b 191
##STR00330## 412.1 0.58 Example 52 192 ##STR00331## 412.1 0.58
Example 52 193 ##STR00332## 502.1 0.65 Example 19 194 ##STR00333##
441.1 0.75 Example 1b 195 ##STR00334## 402.1 0.55 Example 1b 196
##STR00335## 387.1 0.64 Examples 1, 5 197 ##STR00336## 410.3 0.6
Example 68 198 ##STR00337## 472.3 0.66 Examples 1, 10 199
##STR00338## 409.1 0.64 Example 21 200 ##STR00339## 484.3 0.59
Example 19 201 ##STR00340## 441.1 0.73 Example 53 202 ##STR00341##
441.1 0.73 Example 53 203 ##STR00342## 417.3 0.46 Example 85 204
##STR00343## 454.2 0.69 Example 53 205 ##STR00344## 468.1 0.72
Example 53 206 ##STR00345## 549.4 0.67 Example 76 207 ##STR00346##
549.4 0.68 Example 76 208 ##STR00347## 418.3 0.52 Example 75 209
##STR00348## 431.3 0.47 Example 85 210 ##STR00349## 364.2 0.47
Example 85 211 ##STR00350## 406.3 0.53 Example 70 212 ##STR00351##
282.9/ 284.9 0.85 Example 56 213 ##STR00352## 494.2 0.85 Example 53
214 ##STR00353## 454.2 0.71 Example 53 215 ##STR00354## 457.2 0.46
Example 76 216 ##STR00355## 423.2 0.45 Example 76 217 ##STR00356##
457.2 0.47 Example 76 218 ##STR00357## 423.3 0.45 Example 76 219
##STR00358## 427.1 0.7 Example 54 220 ##STR00359## 427.1 0.7
Example 54 221 ##STR00360## 451.2 0.62 Example 60 222 ##STR00361##
452.1 0.76 Example 60 223 ##STR00362## 451.1 0.63 Example 61 224
##STR00363## 460 0.54 Example 33 225 ##STR00364## 505.2 0.64
Example 62 226 ##STR00365## 432.1 0.41 Example 63 227 ##STR00366##
433.1 0.45 Example 63 228 ##STR00367## 417.2 0.51 Example 77 229
##STR00368## 418.2 0.56 Example 77 230 ##STR00369## 464.1/ 466.1
0.44 Example 57 231 ##STR00370## 431.2 0.49 Example 34 232
##STR00371## 417.2 0.47 Example 61 233 ##STR00372## 431.2 0.47
Example 64 234 ##STR00373## 426.2 0.62 Example 54 235 ##STR00374##
440.2 0.62 Example 54 236 ##STR00375## 454.2 0.64 Example 54 237
##STR00376## 426.2 0.62 Example 54 238 ##STR00377## 440.2 0.61
Example 54 239 ##STR00378## 454.2 0.62 Example 54 240 ##STR00379##
457.2 0.6 Example 35 241 ##STR00380## 445.2 0.54 Example 36 242
##STR00381## 432.2 0.56 Example 75 243 ##STR00382## 475.2 0.51
Example 86 244 ##STR00383## 459.2 0.54 Example 78
245 ##STR00384## 495.2 0.57 Examples 6, 85 246 ##STR00385## 431.2
0.51 Examples 77, 85 247 ##STR00386## 473.2 0.57 Example 78 248
##STR00387## 431.2 0.58 Example 77 249 ##STR00388## 445.2 0.59
Example 77 250 ##STR00389## 432.2 0.64 Example 77 251 ##STR00390##
446.2 0.66 Example 77 252 ##STR00391## 489.3 0.57 Example 77, 86
253 ##STR00392## 450.2 0.58 Example 87 254 ##STR00393## 556.2 0.61
Example 79 255 ##STR00394## 485.3 0.63 Example 37 256 ##STR00395##
444.2 0.51 (C18 column), 10.35 (chiral column) Example 24 257
##STR00396## 444.2 0.51 (C18 column), 17.44 (chiral column) Example
24 258 ##STR00397## 508.2 0.63 Example 80 259 ##STR00398## 494.2
0.6 Example 81 260 ##STR00399## 434.2 0.55 Example 38 261
##STR00400## 417.2 0.49 Example 77 262 ##STR00401## 417.2 0.49
Example 77 263 ##STR00402## 418.2 0.54 Example 77 264 ##STR00403##
418.2 0.54 Example 77 265 ##STR00404## 416.2 0.52 Example 82 266
##STR00405## 486 0.7 Example 28 267 ##STR00406## 451.1 0.65 Example
84 268 ##STR00407## 534.3 0.62 Example 83 269 ##STR00408## 434.1
0.57 Example 25 270 ##STR00409## 534.3 0.64 Example 84 271
##STR00410## 550.3 0.62 Example 84 272 ##STR00411## 522.2 0.63
Example 84 273 ##STR00412## 500.3 0.58 Example 82, 83 274
##STR00413## 474.3 0.56 Example 80, 82 275 ##STR00414## 417.2 0.5
Example 84, Intermediate D 276 ##STR00415## 474.3 0.48 Example 80,
Intermediate D 277 ##STR00416## 500.3 0.5 Example 83, Intermediate
D 278 ##STR00417## 500.1 0.49 Example 84, Intermediate D 279
##STR00418## 488.1 0.48 Example 84, Intermediate D 280 ##STR00419##
520.1/ 522 0.59 Example 71 281 ##STR00420## Example 71 282
##STR00421## 339 0.54 Example 39 283 ##STR00422## 492.2 0.57
Example 40 284 ##STR00423## 444.2/ 446.2 0.54 Example 50 285
##STR00424## 502.2/ 504.2 0.56 Example 50 286 ##STR00425## 494.2/
496.1 0.61 Example 41 287 ##STR00426## 488 0.51 Example 67,
Intermediate D 288 ##STR00427## 528.3 0.53 Example 42 289
##STR00428## 528.3 0.53 Example 42 290 ##STR00429## 562.3 0.7
Example 43 291 ##STR00430## 522/ 524 0.62 Example 67 292
##STR00431## 554.1 0.61 Example 44 293 ##STR00432## 506 0.6 Example
39 and Example 67 294 ##STR00433## 506/ 508 0.62 Example 67,
Intermediate I 295 ##STR00434## 576.2 0.78 Example 45 296
##STR00435## 451.2 0.65 Example 46 297 ##STR00436## 479.3 0.72
Example 46 298 ##STR00437## 590.5 0.71 Example 47 299 ##STR00438##
590.5 0.71 Example 47 300 ##STR00439## 416 0.47 Example 1b,
Intermediate D 301 ##STR00440## 528.4 0.53 Example 48 302
##STR00441## 562.4 0.67 Example 49 303 ##STR00442## 402 0.48
Example 1b, Intermediate D 304 ##STR00443## 402 0.48 Example 1b,
Intermediate D 305 ##STR00444## 518.4 0.511 Example 2 306
##STR00445## 516.5 0.653 Example 2 307 ##STR00446## 518.4 0.547
Example 66 308 ##STR00447## 556 0.73 Example 2 309 ##STR00448##
556.4 0.675 Example 2 310 ##STR00449## 513.2 0.563 Example 93 311
##STR00450## 430.3 0.48 Example 29 312 ##STR00451## 448.2 0.62
Example 30 313 ##STR00452## 434.2 0.5 Example 31 314 ##STR00453##
492.3 0.6 Example 1b, Intermediate I 315 ##STR00454## 534.1 0.64
Example 1b, Intermediate W 316 ##STR00455## 301/ 303 0.86 Example
65 317 ##STR00456## 502.3/ 504.3 0.49 Example 58 318 ##STR00457##
452 0.59 Example 1b 319 ##STR00458## 554.1 0.59 Example 1b,
Intermediate I 320 ##STR00459## 564.4/ 566.3 0.65 Example 72 321
##STR00460## 522.1/ 524.0 0.708 Example 74 322 ##STR00461## 552.0/
554.1 0.589 Example 1b, Intermediate I 323 ##STR00462## 522.2/
524.1 0.672 Example 73 324 ##STR00463## 504.1/ 506.1 0.624 Example
1b, Intermediate I 325 ##STR00464## 522.1/ 524.1 0.724 Example 74
326 ##STR00465## 540.2/ 542.2 0.605 Example 1b, Intermediate I 327
##STR00466## 522.1/ 523.9 0.675 Example 73 328 ##STR00467## 516.5
0.55 (C18 column), 9.743 (chiral column) Example 67, Intermediate J
329 ##STR00468## 516.5 0.55 Example XL-1, Intermediate J 330
##STR00469## 516.5 0.55 Example XL-1, Intermediate J 331
##STR00470## 580.1 0.59 Example 1b, Intermediate I 332 ##STR00471##
566.2 0.64 Example 89 333 ##STR00472## 534.1 0.62 Example 90 334
##STR00473## 548.2 0.65 Example 90 335 ##STR00474## 559 0.59
Example 90 336 ##STR00475## 541.3 0.55 Example 90 337 ##STR00476##
560.1 0.73 Example 48 338 ##STR00477## 571.2 0.65 Example 48, 94
339 ##STR00478## 517.2 0.576 Example 94 340 ##STR00479## 531.2
0.595 Example 94 341 ##STR00480## 459.2 0.547 Example 94 342
##STR00481## 501.2 0.627 Example 94 343 ##STR00482## 543.3 0.692
Example 1B 344 ##STR00483## 499.1 0.531 Example 1B 345 ##STR00484##
441.1 0.502 Example 1B 346 ##STR00485## 580.3 0.64 Example 30, 92
347 ##STR00486## 552 0.63 Example 30 348 ##STR00487## 582/ 584 0.58
Example 92 349 ##STR00488## 512.2 0.47 Example 91 350 ##STR00489##
504 0.45 Example 91 351 ##STR00490## 513 0.6 Example 1b, 7 352
##STR00491## 520.1 0.72 Example 1b, 7 353 ##STR00492## 483.2 0.56
Example 1b, 7 354 ##STR00493## 490 0.69 Example 1b, 7 355
##STR00494## 506 0.78 Example 1b, 7 356 ##STR00495##
The following compounds were made using procedures outlined
above:
Compound/Ex. 357:
4-((5'-chloro-5-fluoro-2'-((1r,4r)-4-hydroxycyclohexylamino)-2,4'-bipyrid-
in-6-ylamino)methyl)tetrahydro-2H-pyran-4-carbonitrile
##STR00496##
[0663] M+1 (LC/MS): 460.1; Retention Time (min. LC/MS): 0.62.
[0664] .sup.1H NMR (400 MHz, METHANOL-d4) .delta. ppm 1.29-1.42 (m,
3H) 1.59-1.71 (m, 2H) 1.75-1.80 (m, 1H) 1.80-1.83 (m, 1H) 1.88-1.96
(m, 2H) 1.96-2.02 (m, 2H) 2.02-2.13 (m, 1H) 3.46-3.60 (m, 4H) 3.72
(s, 2H) 3.86 (m, J=12.13, 2.35 Hz, 2H) 6.95 (dd, J=8.02, 2.93 Hz,
1H) 7.10 (s, 1H) 7.32 (dd, J=10.96, 8.22 Hz, 1H) 7.92 (s, 1H).
Compound/Ex. 358:
4-((5'-chloro-2'-((1R,4r)-4-((R)-1-methoxypropan-2-ylamino)cyclohexylamin-
o)-2,4'-bipyridin-6-ylamino)methyl)tetrahydro-2H-pyran-4-carbonitrile
##STR00497##
[0665] Compound/Ex. 359:
4-((5'-chloro-2'-((1r,4r)-4-hydroxycyclohexylamino)-2,4'-bipyridin-6-ylam-
ino)methyl)tetrahydro-2H-pyran-4-carbonitrile
##STR00498##
[0667] M+1 (LC/MS): 442.1; Retention Time (min. LC/MS): 0.55.
[0668] 1H NMR (400 MHz, METHANOL-d4) .delta. ppm 1.29-1.42 (m, 4H)
1.58-1.70 (m, 2H) 1.75-1.84 (m, 2H) 1.87-2.04 (m, 4H) 3.45-3.60 (m,
4H) 3.66 (s, 2H) 3.86 (m, J=12.13, 2.74 Hz, 2H) 6.66 (d, J=8.22 Hz,
1H) 6.88 (d, J=7.43 Hz, 1H) 7.07 (s, 1H) 7.46-7.53 (m, 1H) 7.92 (s,
1H).
Compound/Ex. 360:
4-((5'-chloro-2'-((1r,4r)-4-(ethylamino)cyclohexylamino)-2,4'-bipyridin-6-
-ylamino)methyl)tetrahydro-2H-pyran-4-carbonitrile
##STR00499##
[0670] M+1 (LC/MS): 469.2; Retention Time (min. LC/MS): 0.55.
[0671] .sup.1H NMR (400 MHz, METHANOL-d4) .delta. ppm 1.32 (t,
J=7.24 Hz, 3H) 1.49 (br. s., 4H) 1.66-1.82 (m, 2H) 1.84-1.99 (m,
2H) 2.22 (d, J=12.52 Hz, 4H) 3.11 (t, J=7.24 Hz, 3H) 3.56-3.72 (m,
3H) 3.76 (s, 2H) 3.87-4.06 (m, 2H) 6.81 (d, J=8.61 Hz, 1H) 6.96 (d,
J=6.65 Hz, 1H) 7.06 (s, 1H) 7.54-7.69 (m, 1H) 8.06 (s, 1H).
Compound/Ex. 361:
4-((5'-chloro-2'-((1r,4r)-4-(dimethylamino)cyclohexylamino)-2,4'-bipyridi-
n-6-ylamino)methyl)tetrahydro-2H-pyran-4-carbonitrile
##STR00500##
[0673] M+1 (LC/MS): 469.2; Retention Time (min. LC/MS): 0.52
[0674] .sup.1H NMR (400 MHz, METHANOL-d4) .delta. ppm 1.39-1.58 (m,
2H) 1.64-1.83 (m, 4H) 1.90 (dd, J=13.50, 1.76 Hz, 2H) 2.10-2.35 (m,
4H) 2.87 (s, 6H) 3.57-3.72 (m, 3H) 3.76 (s, 2H) 3.96 (ddd, J=9.98,
2.35, 2.15 Hz, 2H) 6.82 (d, J=7.83 Hz, 1H) 6.97 (d, 6.65 Hz, 1H)
7.06 (s, 1H) 7.55-7.77 (m, 1H) 8.07 (s, 1H).
Compound/Ex. 362:
4-((5'-chloro-2'-((1r,4r)-4-(2-(trifluoromethoxy)ethylamino)cyclohexylami-
no)-2,4'-bipyridin-6-ylamino)methyl)tetrahydro-2H-pyran-4-carbonitrile
##STR00501##
[0676] M+1 (LC/MS): 553.3; Retention Time (min. LC/MS): 0.58.
Compound/Ex. 363:
4-((5'-chloro-2'-((1r,4r)-4-(tetrahydro-2H-pyran-4-ylamino)cyclohexylamin-
o)-2,4'-bipyridin-6-ylamino)methyl)tetrahydro-2H-pyran-4-carbonitrile
##STR00502##
[0678] M+1 (LC/MS): 525.1; Retention Time (min. LC/MS): 0.54.
[0679] .sup.1H NMR (400 MHz, METHANOL-d4) ppm 1.38-1.82 (m, 8H)
1.85-1.95 (m, 2H) 1.96-2.06 (m, 2H) 2.15-2.26 (m, 4H) 3.40-3.56 (m,
3H) 3.58-3.73 (m, 3H) 3.75 (s, 2H) 3.90-4.10 (m, 4H) 6.71-6.80 (m,
1H) 6.94 (s, 2H) 7.54-7.65 (m, 1H) 8.04 (s, 1H).
Compound/Ex. 364:
5'-chloro-N6-((4-fluorotetrahydro-2H-pyran-4-yl)methyl)-N2'-((1r,4r)-4-(2-
-methoxyethylamino)cyclohexyl)-2,4'-bipyridine-2',6-diamine
##STR00503##
[0681] M+1 (LC/MS): 492.2; Retention Time (min. LC/MS): 0.34.
[0682] .sup.1H NMR (400 MHz, METHANOL-d4) .delta. ppm 1.32-1.48 (m,
2H) 1.49-1.65 (m, 2H) 1.72-1.88 (m, 4H) 2.16-2.26 (m, 4H) 3.20-3.27
(m, 2H) 3.42 (s, 2H) 3.60-3.76 (m, 6H) 3.77-3.86 (m, 2H) 6.78 (s,
1H) 6.91 (d, J=7.04 Hz, 1H) 6.96 (d, J=8.61 Hz, 1H) 7.76 (t, J=8.02
Hz, 1H) 8.06 (s, 1H).
Compound/Ex. 365
##STR00504##
[0683] Compound/Ex. 366:
4-((5'-chloro-2'-((1r,4r)-4-(diethylamino)cyclohexylamino)-2,4'-bipyridin-
-6-ylamino)methyl)tetrahydro-2H-pyran-4-carbonitrile
##STR00505##
[0685] M+1 (LC/MS): 497.2; Retention Time (min. LC/MS): 0.58.
Compound/Ex. 367:
2-((5'-chloro-5-fluoro-2'-((1R,4r)-4-((R)-1-methoxypropan-2-ylamino)cyclo-
hexylamino)-2,4'-bipyridin-6-ylamino)methyl)propane-1,3-diol
##STR00506##
[0687] M+1 (LC/MS): 496.2; Retention Time (min. LC/MS): 0.49.
Biological Methods
[0688] Cdk9/cyclinT1 IMAP Protocol
[0689] The biological activity of the compounds of the invention
can be determined using the assay described below.
[0690] Cdk9/cyclinT1 is purchased from Millipore, cat #14-685. The
final total protein concentration in the assay 4 nM. The
5TAMRA-cdk7tide peptide substrate,
5TAMRA-YSPTSPSYSPTSPSYSTPSPS-COOH, is purchased from Molecular
Devices, cat#R7352. The final concentration of peptide substrate is
100 nM. The ATP substrate (Adenosine-5'-triphosphate) is purchased
from Roche Diagnostics, cat#1140965. The final concentration of ATP
substrate is 6 uM. IMAP (Immobilized Metal Assay for
Phosphochemicals) Progressive Binding reagent is purchased from
Molecular Devices, cat#R8139. Fluorescence polarization (FP) is
used for detection. The 5TAMRA-cdk7tide peptide is phosphorylated
by Cdk9/cyclinT1 kinase using the ATP substrate. The
Phospho-5TAMRA-cdk7tide peptide substrate is bound to the IMAP
Progressive Binding Reagent. The binding of the IMAP Progressive
Binding Reagent changes the fluorescence polarization of the
5TAMRA-cdk7tide peptide which is measured at an excitation of 531
nm and FP emission of 595 nm. Assays are carried out in 100 mM
Tris, pH=7.2, 10 mM MgCl2, 0.05% NaN3, 0.01% Tween-20, 1 mM
dithiothreitol and 2.5% dimethyl sulfoxide. IMAP Progressive
Binding Reagent is diluted 1:800 in 100% 1.times. Solution A from
Molecular Devices, cat#R7285.
[0691] General protocol is as follows: To 10 ul of cdk9/cyclinT1,
0.5 ul of test compound in dimethyl sulfoxide is added.
5TAMRA-cdk7tide and ATP are mixed. 10 ul of the 5TAMRA-cdk7tide
iATP mix is added to start the reaction. The reaction is allowed to
proceed for 4.5 hrs. 60 uL of IMAP Progressive Binding Reagent is
added. After >1 hr of incubation, plates are read on the
Envision 2101 from Perkin-Elmer. The assay is run in a 384-well
format using black Corning plates, cat#3573.
Cdk9/cyclinT1 Alpha Screen Protocol
[0692] Full length wild type Cdk9/cyclin T1 is purchased from
Invitogen, cat#PV4131. The final total protein concentration in the
assay 1 nM. The cdk7tide peptide substrate,
biotin-GGGGYSPTSPSYSPTSPSYSPTSPS-OH, is a custom synthesis
purchased from the Tufts University Core Facility. The final
concentration of cdk7tide peptide substrate is 200 nM. The ATP
substrate (Adenosine-5'-triphosphate) is purchased from Roche
Diagnostics. The final concentration of ATP substrate is 6 uM.
Phospho-Rpb1 CTD (ser2/5) substrate antibody is purchased from Cell
Signaling Technology. The final concentration of antibody is 0.67
ug/ml. The Alpha Screen Protein A detection kit containing donor
and acceptor beads is purchased from PerkinElmer Life Sciences. The
final concentration of both donor and acceptor beads is 15 ug/ml.
Alpha Screen is used for detection. The biotinylated-cdk7tide
peptide is phosphorylated by cdk9/cyclinT1 using the ATP substrate.
The biotinylated-cdk7tide peptide substrate is bound to the
streptavidin coated donor bead. The antibody is bound to the
protein A coated acceptor bead. The antibody will bind to the
phosphorylated form of the biotinylated-cdk7tide peptide substrate,
bringing the donor and acceptor beads into close proximity. Laser
irradiation of the donor bead at 680 nm generates a flow of
short-lived singlet oxygen molecules. When the donor and acceptor
beads are in close proximity, the reactive oxygen generated by the
irradiation of the donor beads initiates a
luminescence/fluorescence cascade in the acceptor beads. This
process leads to a highly amplified signal with output in the
530-620 nm range. Assays are carried out in 50 mM Hepes, pH=7.5, 10
mM MgCl2, 0.1% Bovine Serum Albumin, 0.01% Tween-20, 1 mM
Dithiolthreitol, 2.5% Dimethyl Sulfoxide. Stop and detection steps
are combined using 50 mM Hepes, pH=7.5, 18 mM EDTA, 0.1% Bovine
Serum Albumin, 0.01% Tween-20.
[0693] General protocol is as follows: To 5 ul of cdk9/cyclinT1,
0.25 ul of test compound in dimethyl sulfoxide is added. Cdk7tide
peptide and ATP are mixed. 5 ul of the cdk7tide peptide/ATP mix is
added to start the reaction. The reaction is allowed to proceed for
5 hrs. 10 uL of Ab/Alpha Screen beads/Stop-detection buffer is
added. Care is taken to keep Alpha Screen beads in the dark at all
times. Plates are incubated at room temperature overnight, in the
dark, to allow for detection development before being read. The
assay is run is a 384-well format using white polypropylene Greiner
plates.
[0694] The data shown in Table 2 below were generated using one of
the assays described above.
TABLE-US-00002 TABLE 2 Compound CDK9 # in the CYCLINT1 write-up
(IC50) 1 0.007945 2 0.025572 3 0.237603 4 0.009055 5 0.039655 6
0.136417 7 0.024792 8 0.084843 9 0.007945 10 0.018574 11 0.132509
12 0.007945 13 0.007945 14 0.522 15 0.007945 16 0.023617 17
0.02323424 18 0.007945 19 0.007945 20 0.007945 21 0.007945 22
0.007945 23 0.007945 24 0.012416 25 0.008079 26 0.016256 27
0.007945 27 0.007945 29 0.007945 30 0.007945 31 0.044838 32
0.015002 33 0.026973 34 0.048274 35 0.055761 36 0.008906 37
0.007945 38 0.007945 39 0.008896 40 0.007945 41 0.01288 42 0.048069
43 0.007945 44 0.011238 45 46 0.007945 47 0.007945 48 0.00794 49
0.014430 50 0.007945 51 0.017367 52 0.019224 53 0.011128 54
0.023156 55 0.007945 56 0.039262 57 0.032590 58 0.031203 59
0.009128 60 0.007945 61 0.018100 62 0.007945 63 0.007945 64
0.054559 65 0.007945 66 0.017131 67 2.550202 68 0.274123 69
0.154400 70 0.173426 71 0.027388 72 0.114363 73 0.035218 74
0.041585 75 0.013530 76 0.011082 77 0.007945 78 0.024249 79
0.007945 80 0.031705 81 0.054218 82 0.009047 83 0.011615 84
0.014118 85 0.068526 86 0.081460 87 0.068978 88 0.011003 89
2.582156 90 2.960356 91 0.335581 92 0.295616 93 0.928257 94 0.50746
95 1.951420 96 1.276694065 97 0.339265455 98 0.415725004 99
0.679432727 100 0.308717658 101 0.007945668 102 0.120571151 103
0.133698728 104 0.140890633 105 0.0180851 106 0.059240258 107
0.015318231 105 0.084308021 106 0.072890252 107 0.007945668 108
0.007945668 109 0.015815541 110 0.025176571 111 0.030797253 112
0.027282158 113 0.050224047 114 0.007945668 115 0.007945668 116
0.007945668 117 0.123719173 118 0.138887135 119 0.154521231 120
0.045604039 121 10.49437327 122 0.007945668 123 0.042845475 124
0.116276412 125 0.278772642 126 0.033296354 127 0.139053728 128
0.033364795 129 0.390099615 130 0.16902747 131 0.46977199 132
0.014431175 133 0.007945668 134 0.01051692 135 0.142053718 136
0.204223958 137 0.007945668 138 0.521640084 139 0.030140062 140
0.012553271 141 0.204786235 142 0.025611049 143 0.022738812 144
0.015810302 145 0.007945668 146 0.007945668 147 0.019350577 148
15.62589296 149 0.516196192 150 6.512117546 151 0.007945668 152
0.027 153 1.546 154 0.382 155 0.023 156 0.045 157 0.0079 158 0.011
159 1.383 160 0.019 161 0.026 162 0.014 163 0.013 164 0.039 165
0.0079 166 0.027 167 0.018 168 0.037 169 0.009 170 0.044 171 0.218
172 0.015 173 0.062 174 0.029 175 0.024 176 0.021 177 0.013 178
0.103 179 0.544 180 0.01213 181 0.00794 182 0.02111 183 0.00794 184
0.00911 185 0.11048 186 0.00794 187 2.73860 188 0.00794 189 0.00794
190 0.1 191 0.00794 192 0.00794 193 0.00794 194 0.04813 195 0.03556
196 0.81167 197 0.00794 198 0.00794 199 0.00794 200 0.00794 201
0.63424 202 0.01884 203 0.00794 204 0.00794 205 0.01747 206 0.13378
207 0.114147 208 0.00794 209 0.18300 210 0.085970 211 0.02101 212
0.05460 213 0.0142 214 0.04169 215 0.06545 216 0.13825 217 0.03728
218 0.12766 219 0.007945 220 0.007945 221 0.007945 222 0.007945 223
0.007945 224 0.091885 225 0.007945 226 0.007945 227 0.025907 228
0.007945 229 0.007945 230 0.007945 231 0.014853 232 0.007945 233
0.007945 234 0.007945 235 0.007945 236 0.007945 237 0.007945 238
0.013635 239 0.018420 240 0.020961
241 0.06179 242 0.015408 243 0.007945 244 0.078984 245 0.05337 246
0.01154 247 0.02018 248 0.01058 249 0.0318 250 0.02839 251 0.04320
252 0.00794 253 0.00794 254 0.00833 255 0.04232 256 0.00794 257
0.00794 258 0.00794 259 0.00794 260 0.00794 261 0.00794 262 0.00794
263 0.00794 264 0.00794 265 0.03258 266 0.00794 267 0.27007 268
0.008143 269 0.007945 270 0.00794 271 0.00794 272 0.00794 273
0.02293 274 0.03777 275 0.14630 276 0.00893 277 0.00794 278 0.00794
279 0.01310 280 0.0161 281 0.06124 282 0.00794 283 0.001 284 0.001
285 0.002 286 0.001 287 0.003 288 0.003 289 0.004 290 0.002 291
0.001 292 0.002 293 0.001 294 0.001 295 296 0.277 297 0.001 298
0.001 299 0.001 300 0.219 301 0.003 302 0.001 303 1.296 304 6.188
305 0.001 306 0.009 307 0.008 308 0.001 309 0.035 310 0.0003 311
0.001 312 0.0003 313 0.001 314 0.001 315 0.018 316 0.009 317 0.099
318 0.00026 319 0.004 320 0.001 321 0.011 322 0.003 323 0.001 324
0.002 325 0.01 326 0.00049 327 0.001 328 0.001 329 0.001 330 0.002
331 0.001 332 0.001 333 0.004 334 0.001 335 0.00027 336 337 338 339
0.00017 340 0.00023 341 0.00015 342 0.00017 343 0.00031 344 345 346
0.001 347 0.001 348 0.002 349 0.001 350 0.001 351 352 353 354 355
356 357 0.00016 358 0.00017 359 0.00024 360 0.00028 361 0.00030 362
0.00036 363 0.00043 364 0.00063 365 0.00070 366 0.0010 367
0.0031
* * * * *