U.S. patent application number 16/230408 was filed with the patent office on 2019-05-09 for heterocyclic kinase inhibitors.
The applicant listed for this patent is AbbVie Inc.. Invention is credited to Phil Cox, Kevin Cusack, Kristine E. Frank, Michael Friedman, J. Martin Herold, Michael Z. Hoemann, Raymond Huntley, Augustine Osuma, Noel S. Wilson, Xiangdong Xu.
Application Number | 20190135836 16/230408 |
Document ID | / |
Family ID | 54323386 |
Filed Date | 2019-05-09 |
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United States Patent
Application |
20190135836 |
Kind Code |
A1 |
Friedman; Michael ; et
al. |
May 9, 2019 |
HETEROCYCLIC KINASE INHIBITORS
Abstract
This invention provides compounds of Formula (I)
pharmaceutically acceptable salts, pro-drugs, biologically active
metabolites, stereoisomers, and isomers thereof wherein the
variable are defined herein. The compounds of the invention are
useful for treating immunological and oncological conditions.
##STR00001##
Inventors: |
Friedman; Michael;
(Brookline, MA) ; Cox; Phil; (Grayslake, IL)
; Frank; Kristine E.; (Grayslake, IL) ; Hoemann;
Michael Z.; (Marlborough, MA) ; Osuma; Augustine;
(Lindenhurst, IL) ; Wilson; Noel S.; (Kenosha,
WI) ; Xu; Xiangdong; (Buffalo Grove, IL) ;
Cusack; Kevin; (Holden, MA) ; Huntley; Raymond;
(Millbury, MA) ; Herold; J. Martin; (Cambridge,
MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AbbVie Inc. |
North Chicago |
IL |
US |
|
|
Family ID: |
54323386 |
Appl. No.: |
16/230408 |
Filed: |
December 21, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15304131 |
Oct 14, 2016 |
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PCT/CN2015/076766 |
Apr 16, 2015 |
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16230408 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
Y02A 50/463 20180101;
Y02A 50/30 20180101; Y02A 50/401 20180101; A61P 17/06 20180101;
A61P 19/02 20180101; A61P 13/12 20180101; A61P 37/02 20180101; C07D
487/04 20130101; A61K 31/437 20130101; A61P 43/00 20180101; A61P
25/00 20180101; A61P 35/00 20180101; A61P 13/10 20180101; C07D
519/00 20130101; A61P 7/00 20180101; A61P 37/06 20180101; A61P
35/02 20180101; A61P 11/06 20180101; C07D 471/04 20130101; A61P
29/00 20180101; A61P 19/08 20180101; A61P 1/04 20180101; C07D
498/04 20130101 |
International
Class: |
C07D 519/00 20060101
C07D519/00; A61K 31/437 20060101 A61K031/437; C07D 471/04 20060101
C07D471/04; C07D 487/04 20060101 C07D487/04; C07D 498/04 20060101
C07D498/04 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 17, 2014 |
CN |
PCT/CN2014/075560 |
Claims
1. A compound of Formula (I) ##STR00242## wherein U is CR.sup.1 or
N; X is CR.sup.2 or N; Y is CR.sup.3 or N; Z is CR.sup.4 or N;
R.sup.1 is independently H or deuterium; R.sup.2 is H, deuterium,
optionally substituted (C.sub.1-C.sub.3)alkyl, or CF.sub.3; R.sup.3
is H, deuterium or optionally substituted (C.sub.1-C.sub.3)alkyl;
R.sup.4 is H or deuterium; R.sup.5 is --R.sup.501-L-R.sup.502
wherein R.sup.501 is a bond, --O--, --OCH.sub.2--, or optionally
substituted (C.sub.1-C.sub.3)alkylene, L is --C(.dbd.O)--,
--CH.sub.2N(H)C(.dbd.O)--, --N(H)C(.dbd.O)--, or N(H)S(O).sub.2; or
L is a bond and R.sup.302 is --CN; or L is -L.sup.1-L.sup.2 wherein
L.sup.1 is attached to R.sup.501 wherein L.sup.1 is optionally
substituted phenyl, optionally substituted heteroaryl, optionally
substituted saturated or partially saturated heterocyclyl, or
optionally substituted saturated or partially saturated
(C.sub.3-C.sub.7)cycloalkyl and L.sup.2 is a bond,
--CH.sub.2N(R.sup.a)--, --CH.sub.2N(R.sup.a)C(O)--,
--N(R.sup.a)C(O)--, --N(R.sup.a)S(O).sub.2-- or --N(R.sup.a)--; or
L.sup.1 is a saturated or partially saturated heterocyclyl
containing one or more heteroatoms wherein at least one heteroatom
is nitrogen and L.sup.2 is a bond, C(O) or --S(O).sub.2--;
R.sup.502 is H, CF.sub.3, OH, optionally substituted
(C.sub.1-C.sub.6)alkyl, optionally substituted alkenyl, optionally
substituted alkynyl, CN, or optionally substituted
(C.sub.3-C.sub.6)cycloalkenyl; R.sup.6 is optionally substituted
(C.sub.1-C.sub.6)alkyl, optionally substituted
(C.sub.3-C.sub.12)cycloalkyl, optionally substituted phenyl,
optionally substituted heteroaryl, or optionally substituted
heterocyclyl; or R.sup.6 is --R.sup.601--R.sup.602 wherein
R.sup.601 is attached to the --N(H)-- and R.sup.601 is optionally
substituted heteroaryl; R.sup.602 is N(R.sup.a).sub.2, optionally
substituted (C.sub.1-C.sub.6)alkyl, optionally substituted
(C.sub.3-C.sub.6)cycloalkyl, or optionally substituted
heterocyclyl; and R.sup.a is independently H or optionally
substituted (C.sub.1-C.sub.6)alkyl; provided the compound is not
2-(3-{8-[5-(morpholine-4-carbonyl)-pyridin-2-ylamino]-imidazo[1,2-a]pyrid-
ine-6-yl}-phenyl)-N-(5,5,5-trifluoro-4-hydroxy-4-methyl-pent-2-ynyl)-aceta-
mide.
2. The compound according to claim 1 wherein U is CR.sup.1 or N; X
is CR.sup.2 or N; Y is CR.sup.3 or N; Z is CR.sup.4 or N; R.sup.1
is independently H or deuterium; R.sup.2 is H, deuterium,
optionally substituted (C.sub.1-C.sub.3)alkyl, or CF.sub.3; R.sup.3
is H, deuterium or optionally substituted (C.sub.1-C.sub.3)alkyl;
R.sup.4 is H or deuterium; R.sup.5 is --R.sup.501-L-R.sup.502
wherein R.sup.501 is a bond, --O--, --OCH.sub.2--, or optionally
substituted (C.sub.1-C.sub.3)alkylene, L is --C(.dbd.O)--,
--CH.sub.2N(H)C(.dbd.O)--, --N(H)C(.dbd.O)--, or --N(H)S(O).sub.2;
or L is a bond and R.sup.502 is --CN; or L is -L.sup.1-L.sup.2
wherein L.sup.1 is attached to R.sup.501 wherein L.sup.1 is
optionally substituted phenyl, optionally substituted heteroaryl,
optionally substituted saturated or partially saturated
heterocyclyl, or optionally substituted saturated or partially
saturated (C.sub.3-C.sub.6)cycloalkyl and L.sup.2 is a bond,
--CH.sub.2N(R.sup.a)--, --CH.sub.2N(R.sup.a)C(O)--,
--N(R.sup.a)C(O)--, --N(R.sup.a)S(O).sub.2-- or --N(R.sup.a)--; or
L.sup.1 is a saturated or partially saturated heterocyclyl
containing one or more heteroatoms wherein at least one heteroatom
is nitrogen and L.sup.2 is a bond, C(O) or --S(O).sub.2--;
R.sup.502 is H, optionally substituted alkenyl, optionally
substituted alkynyl, CN, or optionally substituted
(C.sub.3-C.sub.6)cycloalkenyl; R.sup.6 is optionally substituted
(C.sub.1-C.sub.6)alkyl, optionally substituted
(C.sub.3-C.sub.12)cycloalkyl, optionally substituted phenyl,
optionally substituted heteroaryl, or optionally substituted
heterocyclyl; and R.sup.a is independently H or optionally
substituted (C.sub.1-C.sub.6)alkyl.
3. The compound of claim 1 wherein L is --C(.dbd.O)--,
--CH.sub.2N(H)C(.dbd.O)--, --N(H)C(.dbd.O)--, or --S(O).sub.2; and
R.sup.502 is H, --CH.dbd.CH.sub.2 or --C.ident.CH; or L is a bond
and R.sup.502 is --CN; or L is -L.sup.1-L.sup.2 wherein L.sup.1 is
attached to R.sup.501 wherein L.sup.1 is optionally substituted
phenyl, optionally substituted heteroaryl or optionally substituted
saturated or partially saturated (C.sub.3-C.sub.6)cycloalkyl and
L.sup.2 is --CH.sub.2N(R.sup.a)--, --CH.sub.2N(R.sup.a)C(O)--,
--N(R.sup.a)C(O)--, --N(R.sup.a)S(O).sub.2-- or --N(R.sup.a)--; or
L.sup.1 is optionally substituted heteroaryl, optionally
substituted azepanyl, optionally substituted azetidinyl, optionally
substituted morpholinyl, optionally substituted oxazepanyl,
optionally substituted piperidinyl, optionally substituted
pyrrolidinyl, optionally substituted tetrahydrofuranyl, or
optionally substituted tetrahydropyranyl, and L.sup.2 is a bond,
C(O) or --S(O).sub.2--.
4. The compound of claim 2 wherein R.sup.6 is optionally
substituted (C.sub.1-C.sub.6)alkyl, optionally substituted phenyl,
optionally substituted bicycle[1.1.1]pentanyl, optionally
substituted 1,2,4 oxadiazolyl, optionally substituted pyrazolyl,
optionally substituted pyridazinyl, optionally substituted
pyridinyl, 4,5-dihydro-1H-benzo[b]azepin-2(3H)-one,
3,4-dihydroquinolin-2(1H)-one, 2H-benzo[b][1,4]oxazin-3(4H)-one, or
6,7-dihydro-4H-pyrazolo [5,1-c][1,4]oxazinyl.
5. The compound of claim 3 wherein R.sup.6 is optionally
substituted with one or more substituents independently selected
from (C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkoxy, optionally
substituted imidazolidinone, or morpholinyl.
6. The compound of claim 4 wherein -L-R.sup.502 forms --CN,
--CH.sub.2N(H)C(.dbd.O)CH.dbd.CH.sub.2, --C(.dbd.O)CH.dbd.CH.sub.2,
--N(H)C(.dbd.O)CH.dbd.CH.sub.2, --N(H)CN, or
--S(O).sub.2CH.dbd.CH.sub.2.
7. The compound according to claim 1 wherein the compound is
N-(3-(8-((4-morpholinophenyl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,5-a]py-
razin-5-yl)benzyl)acrylamide;
N-(3-(8-(bicyclo[1.1.1]pentan-1-ylamino)-1,8a-dihydro-[1,2,4]triazolo[1,5-
-a]pyrazin-5-yl)benzyl)acrylamide;
N-(3-(8-(bicyclo[1.1.1]pentan-1-ylamino)-1,8a-dihydro-[1,2,4]triazolo[1,5-
-a]pyrazin-5-yl)phenyl)acrylamide;
1-(3-(8-((3,4-dimethoxyphenyl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,5-a]p-
yrazin-5-yl)pyrrolidin-1-yl)prop-2-en-1-one;
8-((5-(1-acryloylpyrrolidin-3-yl)-1,8a-dihydro-[1,2,4]triazolo[1,5-a]pyra-
zin-8-yl)amino)-4,5-dihydro-1H-benzo[b]azepin-2(3H)-one;
1-(3-(8-((4-morpholinopyridin-2-yl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,-
5-a]pyrazin-5-yl)pyrrolidin-1-yl)prop-2-en-1-one;
1-(3-(8-((5-methoxypyridazin-3-yl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,5-
-a]pyridin-5-yl)pyrrolidin-1-yl)prop-2-en-1-one;
N-(2-(8-(bicyclo[1.1.1]pentan-1-ylamino)-1,8a-dihydro-[1,2,4]triazolo[1,5-
-a]pyrazin-5-yl)phenyl)acrylamide; 1-((3R)-3-(8-((3,4-di
methoxyphenyl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)pyrr-
olidin-1-yl)prop-2-en-1-one;
1-((3S)-3-(8-((3,4-dimethoxyphenyl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,-
5-a]pyrazin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one;
8-((6-(1-acryloylpiperidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino-
)-4,5-dihydro-1H-benzo[b]azepin-2(3H)-one;
N-(3-(8-((2-oxo-2,3,4,5-tetrahydro-1H-benzo[b]azepin-8-yl)amino)-[1,2,4]t-
riazolo[1,5-a]pyrazin-6-yl)phenyl)acrylamide;
N-(3-(8-((2-oxo-2,3,4,5-tetrahydro-1H-benzo[b]azepin-8-yl)amino)-[1,2,4]t-
riazolo[1,5-a]pyrazin-6-yl)benzyl)acrylamide;
1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)pyrrolidin-1-yl)prop-2-en-1-one;
7-((6-(1-acryloylpyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amin-
o)-3,4-dihydroquinolin-2(1H)-one;
6-((6-(1-acryloylpyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amin-
o)-2H-benzo [b][1,4]oxazin-3(4H)-one;
8-((6-(1-acryloylpyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)amin-
o)-4,5-dihydro-1H-benzo[b]azepin-2(3H)-one;
(S)-8-((6-(1-acryloylpyrrolidin-3-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-4,-
5-dihydro-1H-benzo [b]azepin-2(3H)-one;
(R)-8-((6-(1-acryloylpyrrolidin-3-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-4,-
5-dihydro-1H-benzo [b]azepin-2(3H)-one;
(S)--N-(3,4-dimethoxyphenyl)-6-(1-(vinylsulfonyl)pyrrolidin-3-yl)-[l
1,2,4]triazolo[1,5-a]pyrazin-8-amine;
1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]tr-
iazolo[1,5-a]pyridin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one;
1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)piperidin-1-yl)prop-2-en-1-one;
1-(3-(8-((6-morpholinopyridazin-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-
-6-yl)piperidin-1-yl)prop-2-en-1-one;
1-(3-(8-(methylamino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)pyrrolidin-1-yl)-
prop-2-en-1-one;
1-(3-(8-((2-methoxyethyl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)pyrrol-
idin-1-yl)prop-2-en-1-one;
1-(3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)piperidin-1-yl)prop-2-en-1-one;
1-(3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-
-yl)pyrrolidin-1-yl)prop-2-en-1-one;
1-(3-(8-((6-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)piperidin-1-yl)prop-2-en-1-one;
1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]tr-
iazolo[1,5-a]pyridin-6-yl)piperidin-1-yl)prop-2-en-1-one;
(S)-1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrid-
in-6-yl)pyrrolidin-1-yl)prop-2-en-1-one;
(R)-1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrid-
in-6-yl)pyrrolidin-1-yl)prop-2-en-1-one;
1-(3-(8-((6-morpholinopyridin-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-
-yl)piperidin-1-yl)prop-2-en-1-one;
1-(3-(8-((6-morpholinopyridin-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-
-yl)pyrrolidin-1-yl)prop-2-en-1-one;
1-(3-(8-((3-isopropyl-1,2,4-oxadiazol-5-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)piperidin-1-yl)prop-2-en-1-one;
N-((1R,3S)-3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclohexyl)acrylamide;
N-((1S,3S)-3-(8-((1-methyl-1H-pyrazol-3-yl)methyl)-[1,2,4]triazolo[1,5-a]-
pyridin-6-yl)cyclohexyl)acrylamide;
1-(3-(8-((3-methyl-1,2,4-oxadiazol-5-yl)amino)-[1,2,4]triazolo[1,5-a]pyri-
din-6-yl)piperidin-1-yl)prop-2-en-1-one;
N-((1S,3S)-3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclohexyl)cyanamide;
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)piperidine-1-carbonitrile;
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)pyrrolidine-1-carbonitrile;
1-(4-((6-(1-acryloylpyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)a-
mino)phenyl)-3-methylimidazolidin-2-one;
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)pyrrolidine-1-carbonitrile;
N-((1R,3S)-3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclopentyl)acrylamide; N-((1
S)-3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)cyclopentyl)acrylamide;
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)piperidine-1-carbonitrile;
(S)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,-
4]triazolo[1,5-a]pyridin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one;
(R)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,-
4]triazolo[1,5-a]pyridin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one;
1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-
-yl)pyrrolidin-1-yl)prop-2-en-1-one;
(S)-1-(3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyraz-
in-6-yl)pyrrolidin-1-yl)prop-2-en-1-one;
(R)-1-(3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyraz-
in-6-yl)pyrrolidin-1-yl)prop-2-en-1-one;
(S)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,-
4]triazolo[1,5-a]pyridin-6-yl)piperidin-1-yl)prop-2-en-1-one;
1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)imidazo[1,-
2-b]pyridazin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one; or
(R)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,-
4]triazolo[1,5-a]pyridin-6-yl)piperidin-1-yl)prop-2-en-1-one.
8. The compound according to claim 1 wherein R.sup.5 is
--R.sup.501-L-R.sup.502 wherein R.sup.501 is a bond; L is
-L.sup.1-L.sup.2 wherein L.sup.1 is attached to R.sup.501 wherein
L.sup.1 is optionally substituted saturated or partially saturated
(C.sub.3-C.sub.7)cycloalkyl and L.sup.2 is a
bond-CH.sub.2N(R.sup.a)C(O)--, or --N(R.sup.a)C(O)--; and R.sup.502
is H, CF.sub.3, OH, optionally substituted (C.sub.1-C.sub.6)alkyl,
optionally substituted alkenyl, optionally substituted alkynyl, CN,
or optionally substituted (C.sub.3-C.sub.6)cycloalkenyl.
9. A compound according to claim 8 wherein R.sup.6 is optionally
substituted (C.sub.1-C.sub.6)alkyl, optionally substituted
(C.sub.3-C.sub.12)cycloalkyl, optionally substituted phenyl,
optionally substituted pyrazolyl, optionally substituted
6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazinyl, optionally substituted
4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinyl; or R.sup.6 is
--R.sup.601--R.sup.602 wherein R.sup.601 is attached to the
--N(H)-- and R.sup.601 is optionally substituted pyrazolyl, or
optionally substituted pyridinyl; R.sup.602 is N(R.sup.a).sub.2,
optionally substituted (C.sub.1-C.sub.6)alkyl, optionally
substituted (C.sub.3-C.sub.6)cycloalkyl, optionally substituted
azetidinyl, optionally substituted morpholinyl, optionally
substituted piperidinyl, or optionally substituted
tetrahydropyranyl.
10. The compound according to claim 9 wherein R.sup.1 is H.
11. The compound according to claim 10 wherein X is N or CR.sup.2
wherein R.sup.2 is H, optionally substituted
(C.sub.1-C.sub.3)alkyl, or CF.sub.3.
12. The compound according to claim 11 wherein R.sup.3 is H,
deuterium or optionally substituted (C.sub.1-C.sub.3)alkyl.
13. The compound according to claim 12 wherein U is CH.
14. The compound according to claim 13 wherein X is N.
15. The compound according to claim 1 wherein the compound is
4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)-2-methylbutan-2-ol;
6-cyclohexyl-N-(1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)-[1,2,4]tria-
zolo[1,5-a]pyrazin-8-amine;
6-cyclohexyl-N-(1-(2,2,6,6-tetramethyltetrahydro-2H-pyran-4-yl)-1H-pyrazo-
l-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amine;
6-cyclohexyl-N-(1-(oxetan-3-yl)-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]py-
razin-8-amine;
(1R,4R)-4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H--
pyrazol-1-yl)cyclohexanol;
(1S,4S)-4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H--
pyrazol-1-yl)cyclohexanol;
6-cyclohexyl-N-(1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amine;
(6-cyclohexyl-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5--
a]pyrazin-8-amine;
6-cyclopentyl-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5--
a]pyrazin-8-amine;
6-cyclohexyl-N-(1-isopropyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazi-
n-8-amine;
6-(4,4-dimethylcyclohexyl)-N-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]-
triazolo[1,5-a]pyrazin-8-amine;
N-(1-methyl-1H-pyrazol-4-yl)-6-((1R,4R)-4-methylcyclohexyl)-[1,2,4]triazo-
lo[1,5-a]pyrazin-8-amine;
N-(1-methyl-1H-pyrazol-4-yl)-6-((1S,4S)-4-methylcyclohexyl)-[1,2,4]triazo-
lo[1,5-a]pyrazin-8-amine;
N-(1-methyl-1H-pyrazol-4-yl)-6-((1r,4r)-4-(trifluoromethyl)cyclohexyl)-[1-
,2,4]triazolo[1,5-a]pyrazin-8-amine;
N-(6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazo-
lo[5,1-c][1,4]oxazin-2-amine;
6-cyclohexyl-N-(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)-[-
1,2,4]triazolo[1,5-a]pyridin-8-amine;
6-cyclopentyl-N-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin--
8-amine;
6-cyclopentyl-N-(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazi-
n-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-amine;
1-(4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyraz-
ol-1-yl)piperidin-1-yl)ethanone;
4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)-N-methylpiperidine-1-carboxamide;
(1S,3S)-3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H--
pyrazol-1-yl)cyclohexanol;
(1R,3R)-3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H--
pyrazol-1-yl)cyclohexanol;
(1R,3S)-3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H--
pyrazol-1-yl)cyclohexanol;
(1S,3R)-3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H--
pyrazol-1-yl)cyclohexanol;
(1R,3R)-3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyra-
zin-6-yl)cyclohexanol;
1-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)-2-methylpropan-2-ol;
N-(6-cyclopentyl-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyraz-
olo[5,1-c][1,4]oxazin-2-amine;
1-(6-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyridin-8-yl)amino)pyridin-3-yl-
)piperidin-4-ol;
6-cyclohexyl-N-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazo-
lo[1,5-a]pyrazin-8-amine; (1S,4S)-ethyl
4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)cyclohexanecarboxylate;
6-cyclopentyl-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5--
a]pyrazin-8-amine;
6-cyclohexyl-N-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-
-amine;
cis-4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)--
1H-pyrazol-1-yl)cyclohexanecarboxylic acid; or
3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)propan-1-ol.
16. A method of treating a disease comprising administering a
therapeutically effective amount of a compound of claim 1 to a
patient in need thereof.
17. The method according to claim 16, wherein the disease is
rheumatoid arthritis, juvenile rheumatoid arthritis,
osteoarthritis, Crohn's disease, inflammatory bowel disease,
irritable bowel syndrome, ulcerative colitis, psoriatic arthritis,
psoriasis, ankylosing spondylitis, interstitial cystitis, asthma,
systemic lupus erythematosus, lupus nephritis, B cell chronic
lymphocytic lymphoma, multiple sclerosis, chronic lymphocytic
leukemia, small lymphocytic lymphoma, mantle cell lymphoma, B-cell
non-Hodgkin's lymphoma, activated B-cell like diffuse large B-cell
lymphoma, multiple myeloma, diffuse large B-cell lymphoma,
follicular lymphoma, hairy cell leukemia or Lymphoblastic
lymphoma.
18. A kit comprising a packaged product comprising components with
which to administer a compound of claim 1 for treatment of an
autoimmune disorder.
19. The kit according to claim 18, wherein the packaged product
comprises a compound of claim 1 and instructions for use.
20. A pharmaceutical composition comprising a compound according to
claim 1 and one or more pharmaceutically acceptable excipients.
Description
REFERENCE TO RELATED APPLICATION
[0001] This application claims priority to and the benefit of the
filing date of International Application No. PCT/CN2014/075560,
filed on Apr. 17, 2014, the entire content of which is incorporated
herein by reference.
BACKGROUND OF THE INVENTION
[0002] The protein kinases represent a large family of proteins
that play a central role in the regulation of a wide variety of
cellular processes and maintenance of cellular function. A partial,
non-limiting, list of these kinases include: non-receptor tyrosine
kinases such as the Tec family (BTK, ITK, Tec, ETK/BMX &
RLK/TXK), Janus kinase family (Jak1, Jak2, Jak3 and Tyk2); the
fusion kinases, such as BCR-Abl, focal adhesion kinase (FAK), Fes,
Lck and Syk; receptor tyrosine kinases such as colony stimulating
factor 1 receptor (CSF-1R), epidermal growth factor receptor
(EGFR), the platelet-derived growth factor receptor kinase
(PDGF-R), the receptor kinase for stem cell factor, c-kit, the
hepatocyte growth factor receptor, c-Met, and the fibroblast growth
factor receptor, FGFR3; and serine/threonine kinases such as b-RAF,
mitogen-activated protein kinases (e.g., MKK6) and SAPK2.beta..
Aberrant kinase activity has been observed in many disease states
including benign and malignant proliferative disorders as well as
diseases resulting from inappropriate activation of the immune and
nervous systems. The novel compounds of this invention inhibit the
activity of one or more protein kinases and are, therefore,
expected to be useful in the treatment of kinase-mediated
diseases.
[0003] Bruton's tyrosine kinase (BTK) is a non-receptor tyrosine
kinase with a key role in immunoreceptor signaling (BCR,
Fc.epsilon.R, Fc.gamma.R, DAP12, Dectin-1, GPVI etc) in a host of
hematopoietic cells including B cells, platelets, mast cells,
basophils, eosinophils, macrophages and neutrophils as well as
osteoclasts involved in bone destruction (for reviews, see Brunner
et al., 2005 Histol. Histopathol., 20:945, Mohamed et al., 2009
Immunol. Rev., 228:58). Mutations in BTK are known to lead to
X-linked agammaglobulinemia (XLA) in humans and X-linked
immunodeficiency (Xid) in mice, which are characterized by limited
B-cell production & reduced antibody titers (Lindvall et al.,
2005 Immunol. Rev., 203:200). The combined action of BTK in
multiple cell types makes it an attractive target for autoimmune
disease. BTK is related with sequence homology to other Tec family
kinases (ITK, Tec, ETK/BMX & RLK/TXK).
[0004] In B-lymphocytes, BTK is required for B-cell development and
for Ca.sup.2+ mobilization following of B-cell receptor (BCR)
engagement (Khan et al., 1995 Immunity 3:283; Genevier et al., 1997
Clin. Exp. Immun., 110:286) where it is believed to downstream of
Src family kinases (such as Lyn), Syk & PI3K. BTK has been
shown to be important for both thymus-dependent and
thymus-independent type 2 responses to antigens (Khan et al.,
Immunity 1995; 3; 283). In mast cells, studies using BTK mouse
knock-outs (Hata et al., 1998 J. Exp. Med., 187:1235; Schmidt et
al., 2009 Eur. J. Immun., 39:3228) indicate a role for BTK in
Fc.epsilon.RI induced signaling, histamine release & production
of cytokines such as TNF, IL-2, & IL-4. In platelets, BTK is
important for signaling through the glycoprotein VI (GPVI) receptor
that responds to collagen and has been shown to promote platelet
aggregation and contribute to cytokine production from
fibroblast-like synoviocytes (Hsu et al., 2013 Immun. Letters,
150:97). In monocytes and macrophages, the action of BTK in invoked
in Fc.gamma.RI induced signaling and may also have role in
Toll-Like Receptor-induced cytokine responses including TLR2, TLR4,
TLR8 & TLR9 (Horwood et al., 2003 J. Exp. Med., 197:1603;
Horwood et al., 2006 J. Immunol., 176:3635; Perez de Diego et al.,
2006 Allerg. Clin. Imm., 117:1462; Doyle et al., 2007 J. Biol.
Chem., 282:36959, Hasan et al., 2007 Immunology, 123:239; Sochorava
et al., 2007 Blood, 109:2553; Lee et al., 2008, J. Biol. Chem.,
283:11189).
[0005] Therefore, inhibition of BTK is expected to intervene at
several critical junctions of the inflammatory reactions resulting
in an effective suppression of autoimmune response. As such
diseases involving B-cell receptor activation, antibody-Fc receptor
interactions & GPVI receptor signaling may be modulated by
treatment with BTK inhibitors. BTK inhibition is likely to act on
both the initiation of autoimmune disease by blocking BCR signaling
and the effector phase by abrogation of FcR signaling on
macrophages, neutrophils, basophils, and mast cells. Furthermore,
blocking BTK would provide additional benefit via inhibition of
osteoclast maturation and therefore attenuate the bone erosions
& overall joint destruction associated with rheumatoid
arthritis. Inhibiting BTK may be useful in treating a host of
inflammatory and allergic diseases--for example (but not limited
to), rheumatoid arthritis (RA), systemic lupus erythematosus (SLE),
multiple sclerosis (MS) and type I hypersensitivity reactions such
as allergic rhinitis, allergic conjunctivitis, atopic dermatitis,
allergic asthma and systemic anaphylaxis. For a review on targeting
BTK as a treatment for inflammatory disorders and autoimmunity as
well as leukemias and lymphomas, see Uckun & Qazi, 2010 Expert
Opin. Ther. Pat., 20:1457. Because BTK is highly expressed in
cancers of the hematopoietic system & BTK-dependent signaling
in believed to be disregulated there, BTK inhibitors are expected
to be useful treatments for B-cell lymphomas/leukemias & other
oncologic disease--for example (but not limited to) acute
lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL),
non-Hodgkin's lymphoma (NHL), small lymphocytic lymphoma (SLL), and
acute myeloid leukemia (for review, see Buggy & Elias 2012 Int
Rev Immunol. 31:119). Taken together, BTK inhibitors provide a
strong method to treat a host of inflammatory diseases and
immunological disorders as well as hematologic cancers.
[0006] Colony stimulating factor 1 receptor (CSF-1R) is a
homodimeric, class III receptor tyrosine kinase that is encoded by
the FMS proto-oncogene. It is a 972 amino acid transmembrane
protein characterized by an extracellular ligand-binding domain, a
single transmembrane domain (TM) a juxtamembrane domain (JM), two
intracellular kinase domains (TK1 and TK2), divided by a kinase
insert domain (KI), and a c-Terminal domain, UniProt Entry P07333
(Patel et al 2009 Current Topics in Medicinal Chemistry 9:599).
Binding of CSF-1 to the extracellular domain of CSF-1R stabilizes
receptor dimerization, induces trans-autophosphorylation of the
intracellular domain, and activates downstream cytoplasmic
signaling. Small molecule inhibitors of CSF-1R active site block
receptor autophosphorylation and subsequently block the signals
that control the survival, expression, proliferation and
differentiation of macrophages.
[0007] CSF-1R regulates monocyte survival, proliferation and
differentiation as well as macrophage migration (Pixley et al 2004
TRENDS in Cell Biology, 14:628). The natural ligands for CSF-1R
have been identified as CSF-1 and IL-34. CSF-1R is expressed in
myelomonocytic lineage cell, including hemopoietic progenitors,
tissue macrophages, immature B cells, which are implicated in RA
pathogenesis (Hamilton 2008 Nature Reviews Immunology 8:533).
Activation of CSF-1R is known to play a role in a number of
diseases including, but not limited to, RA, Chrohn's disease,
ulcerative colitis, ankylosing spondylitis and cancer (Toh et al
2014 Arthritis & Rheumatology 66:2989: Hume et al 2012 Blood
119:1810 and Campbell et al 2000 Journal of Leukocyte Biology
68:144). The natural ligands, CSF-1 and IL-34, are highly expressed
in the synovial membrane of RA patients, and CSF-1 levels are
increased in the serum and synovial fluid of RA patients and
associated with disease activity (Firestein et al 1988 Journal of
Experimental Medicine 168:1573; Kawaji et al 1995 Nippon Ika
Daigaku Zasshi 62:260; Ritchlin et al 1994 Scand. J. Immunol.
40:292; Takei et al 2000 J. Rheumatol. 27:894; Hwang et al 2012
Arthritis Research & Therapy 14:R14 and Chemel et al 2012 Ann.
Rheum. Dis. 71:150).
[0008] Monocytes derived from RA patients express elevated levels
of Fc.gamma.R I, IIa and IIIa, increased CD14 and oxygen radicals,
and reduced HLA-DR (Shinohara et al 1992 J. Rheumatol. 19:211).
This monocyte phenotype can be produced in vitro and in vivo with
recombinant CSF-1 (Weiner et al 1994 Cancer Res. 54:4084).
Therefore, CSF-1 may drive the recruitment, differentiation and
survival of RA synovial macrophages, and in the local proliferation
of myeloid progenitors. Further, CSF-1 primes macrophages for
greater expression of TNF and other cytokines (Hanamura 1997
Immunopharmacology 37:15). It has been proposed that CSF-1R is
involved in a positive feedback loop for chronic inflammation where
macrophages secrete TNF and IL-1 that induce stromal cell
expression of CSF-1, leading to further expansion of macrophages
and additional expression of TNF and IL-1 (Hamilton 1993 Lancet
342:536).
[0009] CSF-1 deficient mice have been reported to be resistant to
collagen induced arthritis and in a murine model of CIA, CSF-1 was
shown to exacerbate disease while the neutralizing anti-CSF-1
antibody ameliorated disease (Campbell et al 2000 Journal of
Leukocyte Biology 68:144). An anti-CSF-1R monoclonal antibody was
also shown to be efficacious in 2 different animal models for RA
(Toh et al 2014 Arthritis & Rheumatology 66:2989). Small
molecule inhibitor, GW2580, has been shown to inhibit LPS-induced
TNF production in mice (Conway et al 2005 PNAS 102:16078).
Additionally, there are several reports of non-selective small
molecule CSF-1R inhibitors that have shown efficacy in preclinical
disease models for arthritis (Paniagua et al 2006 J. Clin. Invest.
116:2633; Conway et al 2008 J. Pharmacol. Exp. Ther. 326:41; Ohno
et al 2008 Eur. J. Immunol. 38:283; Paniagua et al 2010 Arthritis
Res. Ther. 12:R32 and Madan et al 2012 J. Imuunol. 189:4123).
[0010] Tumor associated macrophages have been associated with poor
prognosis in various cancers and are involved in the promotion of
angiogenesis, invasion and metastasis (Bingle et al 2002 J. Pathol.
196:254; Pollard 2004 Nat. Rev. Cancer 4:71 and Lewis et al 2006
Cancer Res. 66:605). CSF-1 deficient mice with MMTV-PyMT transgenic
tumors exhibited decreased macrophage recruitment and a decreased
rate of tumor progression to metastasis (Lewis et al 2006 Cancer
Res. 66:605). Mammary epithelial expression of CSF-1 was shown to
restore macrophage infiltration and metastatic tumor vasculature
was characterized, and the induction of vasculature, was shown to
be regulated by Tumor-associated macrophages (TAMs) (Lin et al 2001
J. Exp. Med. 193:727). Human mammary tumor xenografts in mice with
CSF-1 antisense oligonucleotide (ODN-196) or small interfering RNAs
CSF-1 siRNA and FMS siRNA) down-regulated target proteins and
suppressed mammary tumor growth (Biswas et al 2008 J. Immunol.
180:2011). Expression of FMS in breast cancer has been linked to
poor survivability and increased tumor size (Kluger et al 2004
Clin. Cancer Res. 10:173; Lin et al 2001 J. Exp. Med. 193:727; Yee
et al 2000 Anticancer Res. 20:4379).
[0011] CSF-1 antibodies have shown therapeutic potential in
treating solid tumors. Treatment with Anti-CSF01 Fab antibody in an
MCF-7 mammary xenograft mouse model suppressed tumor growth (Paulus
et al 2006 Cancer Res. 66:4349). A small molecule inhibitor,
Ki20227, of CSF-1R suppressed osteolytic bone destruction in a
metastasis model (Ohno 2006 Mol. Cancer Ther. 5:2634). In a
separate study, CSF-1 production was also shown to contribute to
osteoclastogenesis from TAMs and to tumor-associated osteolysis
(Yang 2002 J. Bone Joint Surg. Br. 84:452).
[0012] Therefore inhibition of CSF-1 might be of therapeutic value
in treatment of autoimmune diseases and cancer.
SUMMARY OF THE INVENTION
[0013] In a first embodiment the invention provides a compound of
Formula (I)
##STR00002##
wherein
[0014] U is CR.sup.1 or N;
[0015] X is CR.sup.2 or N;
[0016] Y is CR.sup.3 or N;
[0017] Z is CR.sup.4 or N;
[0018] R.sup.1 is independently H or deuterium;
[0019] R.sup.2 is H, deuterium, optionally substituted
(C.sub.1-C.sub.3)alkyl, or CF.sub.3;
[0020] R.sup.3 is H, deuterium or optionally substituted
(C.sub.1-C.sub.3)alkyl;
[0021] R.sup.4 is H or deuterium;
[0022] R.sup.5 is --R.sup.501-L-R.sup.502 wherein [0023] R.sup.501
is a bond, --O--, --OCH.sub.2--, or optionally substituted
(C.sub.1-C.sub.3)alkylene, [0024] L is --C(.dbd.O)--,
--CH.sub.2N(H)C(.dbd.O)--, --N(H)C(.dbd.O)--, or N(H)S(O).sub.2; or
[0025] L is a bond and R.sup.502 is --CN; or [0026] L is
-L.sup.1-L.sup.2 wherein L is attached to R.sup.501 wherein [0027]
L.sup.1 is optionally substituted phenyl, optionally substituted
heteroaryl, optionally substituted saturated or partially saturated
heterocyclyl, or optionally substituted saturated or partially
saturated (C.sub.3-C.sub.7)cycloalkyl and L.sup.2 is a bond,
--CH.sub.2N(R.sup.a)--, --CH.sub.2N(R.sup.a)C(O)--,
--N(R.sup.a)C(O)--, --N(R.sup.a)S(O).sub.2-- or --N(R.sup.a)--; or
[0028] L.sup.1 is a saturated or partially saturated heterocyclyl
containing one or more heteroatoms wherein at least one heteroatom
is nitrogen and L.sup.2 is a bond, C(O) or --S(O).sub.2--; [0029]
R.sup.502 is H, CF.sub.3, OH, optionally substituted
(C.sub.1-C.sub.6)alkyl, optionally substituted alkenyl, optionally
substituted alkynyl, CN, or optionally substituted
(C.sub.3-C.sub.6)cycloalkenyl;
[0030] R.sup.6 is optionally substituted (C.sub.1-C.sub.6)alkyl,
optionally substituted (C.sub.3-C.sub.12)cycloalkyl, optionally
substituted phenyl, optionally substituted heteroaryl, or
optionally substituted heterocyclyl; or
[0031] R.sup.6 is --R.sup.601--R.sup.602 wherein R.sup.601 is
attached to the --N(H)-- and [0032] R.sup.601 is optionally
substituted heteroaryl; [0033] R.sup.602 is N(R.sup.a).sub.2,
optionally substituted (C.sub.1-C.sub.6)alkyl, optionally
substituted (C.sub.3-C.sub.6)cycloalkyl, or optionally substituted
heterocyclyl; and
[0034] R.sup.a is independently H or optionally substituted
(C.sub.1-C.sub.6)alkyl;
provided the compound is not
2-(3-{8-[5-(morpholine-4-carbonyl)-pyridin-2-ylamino]-imidazo[1,2-a]pyrid-
ine-6-yl}-phenyl)-N-(5,5,5-trifluoro-4-hydroxy-4-methyl-pent-2-ynyl)-aceta-
mide.
[0035] In a second embodiment the invention provides a compound
according to the first embodiment wherein
[0036] U is CR.sup.1 or N;
[0037] X is CR.sup.2 or N;
[0038] Y is CR.sup.3 or N;
[0039] Z is CR.sup.4 or N;
[0040] R.sup.1 is independently H or deuterium;
[0041] R.sup.2 is H, deuterium, optionally substituted
(C.sub.1-C.sub.3)alkyl, or CF.sub.3;
[0042] R.sup.3 is H, deuterium or optionally substituted
(C.sub.1-C.sub.3)alkyl;
[0043] R.sup.4 is H or deuterium;
[0044] R.sup.5 is --R.sup.501-L-R.sup.502 wherein [0045] R.sup.501
is a bond, --O--, --OCH.sub.2--, or optionally substituted
(C.sub.1-C.sub.3)alkylene, [0046] L is --C(.dbd.O)--,
--CH.sub.2N(H)C(.dbd.O)--, --N(H)C(.dbd.O)--, or --N(H)S(O).sub.2;
or [0047] L is a bond and R.sup.502 is --CN; or [0048] L is
-L.sup.1-L.sup.2 wherein L.sup.1 is attached to R.sup.501 wherein
[0049] L.sup.1 is optionally substituted phenyl, optionally
substituted heteroaryl, optionally substituted saturated or
partially saturated heterocyclyl, or optionally substituted
saturated or partially saturated (C.sub.3-C.sub.6)cycloalkyl and
L.sup.2 is a bond, --CH.sub.2N(R.sup.a)--,
--CH.sub.2N(R.sup.a)C(O)--, --N(R.sup.a)C(O)--,
--N(R.sup.a)S(O).sub.2-- or --N(R.sup.a)--; or [0050] L.sup.1 is a
saturated or partially saturated heterocyclyl containing one or
more heteroatoms wherein at least one heteroatom is nitrogen and
L.sup.2 is a bond, C(O) or --S(O).sub.2--; [0051] R.sup.502 is H,
optionally substituted alkenyl, optionally substituted alkynyl, CN,
or optionally substituted (C.sub.3-C.sub.6)cycloalkenyl;
[0052] R.sup.6 is optionally substituted (C.sub.1-C.sub.6)alkyl,
optionally substituted (C.sub.3-C.sub.12)cycloalkyl, optionally
substituted phenyl, optionally substituted heteroaryl, or
optionally substituted heterocyclyl; and
[0053] R.sup.a is independently H or optionally substituted
(C.sub.1-C.sub.6)alkyl.
[0054] In a third embodiment the invention provides a compound
according to any of the foregoing embodiments wherein [0055] L is
--C(.dbd.O)--, --CH.sub.2N(H)C(.dbd.O)--, --N(H)C(.dbd.O)--, or
--S(O).sub.2; and R.sup.502 is H, --CH.dbd.CH.sub.2 [0056] or
--C.ident.CH; or [0057] L is a bond and R.sup.502 is --CN; or
[0058] L is -L.sup.1-L.sup.2 wherein L.sup.1 is attached to
R.sup.501 wherein [0059] L.sup.1 is optionally substituted phenyl,
optionally substituted heteroaryl or optionally substituted
saturated or partially saturated (C.sub.3-C.sub.6)cycloalkyl and
L.sup.2 is --CH.sub.2N(R.sup.a)--, --CH.sub.2N(R.sup.a)C(O)--,
--N(R.sup.a)C(O)--, --N(R.sup.a)S(O).sub.2-- or --N(R.sup.a)--; or
[0060] L.sup.1 is optionally substituted heteroaryl, optionally
substituted azepanyl, optionally substituted azetidinyl, optionally
substituted morpholinyl, optionally substituted oxazepanyl,
optionally substituted piperidinyl, optionally substituted
pyrrolidinyl, optionally substituted tetrahydrofuranyl, or
optionally substituted tetrahydropyranyl, and L.sup.2 is a bond,
C(O) or --S(O).sub.2--.
[0061] In a fourth embodiment the invention provides a compound
according to any of the foregoing embodiments wherein R.sup.6 is
optionally substituted (C.sub.1-C.sub.6)alkyl, optionally
substituted phenyl, optionally substituted bicycle[1.1.1]pentanyl,
optionally substituted 1,2,4 oxadiazolyl, optionally substituted
pyrazolyl, optionally substituted pyridazinyl, optionally
substituted pyridinyl, 4,5-dihydro-1H-benzo[b]azepin-2(3H)-one,
3,4-dihydroquinolin-2(1H)-one, 2H-benzo[b][1,4]oxazin-3(4H)-one, or
6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazinyl.
[0062] In a fifth embodiment the invention provides a compound
compound according to any of the foregoing embodiments wherein
R.sup.6 is optionally substituted with one or more substituents
independently selected from (C.sub.1-C.sub.3)alkyl,
(C.sub.1-C.sub.3)alkoxy, optionally substituted imidazolidinone, or
morpholinyl.
[0063] In a sixth embodiment the invention provides a compound
compound according to any of the foregoing embodiments wherein
-L-R.sup.502 forms --CN, --CH.sub.2N(H)C(.dbd.O)CH.dbd.CH.sub.2,
--C(.dbd.O)CH.dbd.CH.sub.2, --N(H)C(.dbd.O)CH.dbd.CH.sub.2,
--N(H)CN, or --S(O).sub.2CH.dbd.CH.sub.2.
[0064] In a seventh embodiment the invention provides a compound
according to compound according to any of the foregoing embodiments
wherein the compound is [0065]
N-(3-(8-((4-morpholinophenyl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,5-a]py-
razin-5-yl)benzyl)acrylamide; [0066]
N-(3-(8-(bicyclo[1.1.1]pentan-1-ylamino)-1,8a-dihydro-[1,2,4]triazolo[1,5-
-a]pyrazin-5-yl)benzyl)acrylamide; [0067]
N-(3-(8-(bicyclo[1.1.1]pentan-1-ylamino)-1,8a-dihydro-[1,2,4]triazolo[1,5-
-a]pyrazin-5-yl)phenyl)acrylamide; [0068]
1-(3-(8-((3,4-dimethoxyphenyl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,5-a]p-
yrazin-5-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0069]
8-((5-(1-acryloylpyrrolidin-3-yl)-1,8a-dihydro-[1,2,4]triazolo[1,5-a]pyra-
zin-8-yl)amino)-4,5-dihydro-1H-benzo[b]azepin-2(3H)-one; [0070]
1-(3-(8-((4-morpholinopyridin-2-yl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,-
5-a]pyrazin-5-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0071]
1-(3-(8-((5-methoxypyridazin-3-yl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,5-
-a]pyridin-5-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0072]
N-(2-(8-(bicyclo[1.1.1]pentan-1-ylamino)-1,8a-dihydro-[1,2,4]triazolo[1,5-
-a]pyrazin-5-yl)phenyl)acrylamide; [0073]
1-((3R)-3-(8-((3,4-dimethoxyphenyl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,-
5-a]pyrazin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0074]
1-((3S)-3-(8-((3,4-dimethoxyphenyl)amino)-1,8a-dihydro-[1,2,4]triazolo[1,-
5-a]pyrazin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0075]
8-((6-(1-acryloylpiperidine-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amin-
o)-4,5-dihydro-1H-benzo[b]azepin-2(3H)-one; [0076]
N-(3-(8-((2-oxo-2,3,4,5-tetrahydro-1H-benzo[b]azepin-8-yl)amino)-[1,2,4]t-
riaz[1,2,4]triazolo[1,5-a]pyrazin-6-yl)phenyl)acrylamide; [0077]
N-(3-(8-((2-oxo-2,3,4,5-tetrahydro-1H-benzo[b]azepin-8-yl)amino)-[1,2,4]t-
riazolo[1,5-a]pyrazin-6-yl)benzyl)acrylamide; [0078]
1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0079]
7-((6-(1-acryloylpyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amin-
o)-3,4-dihydroquinolin-2(1H)-one; [0080]
6-((6-(1-acryloylpyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amin-
o)-2H-benzo [b][1,4]oxazin-3(4H)-one; [0081]
8-((6-(1-acryloylpyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)amin-
o)-4,5-dihydro-1H-benzo[b]azepin-2(3H)-one; [0082]
(S)-8-((6-(1-acryloylpyrrolidin-3-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-4,-
5-dihydro-1H-benzo [b]azepin-2(3H)-one; [0083]
(R)-8-((6-(1-acryloylpyrrolidin-3-yl)imidazo[1,2-a]pyrazin-8-yl)amino)-4,-
5-dihydro-1H-benzo [b]azepin-2(3H)-one; [0084]
(S)--N-(3,4-dimethoxyphenyl)-6-(1-(vinylsulfonyl)pyrrolidin-3-yl)-[1,2,4]-
triazolo[1,5-a]pyrazin-8-amine; [0085]
1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]tr-
iazolo[1,5-a]pyridin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0086]
1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)piperidin-1-yl)prop-2-en-1-one; [0087]
1-(3-(8-((6-morpholinopyridazin-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-
-6-yl)piperidin-1-yl)prop-2-en-1-one; [0088]
1-(3-(8-(methylamino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)pyrrolidin-1-yl)-
prop-2-en-1-one; [0089]
1-(3-(8-((2-methoxyethyl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)pyrrol-
idin-1-yl)prop-2-en-1-one; [0090]
1-(3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)piperidin-1-yl)prop-2-en-1-one; [0091]
1-(3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-
-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0092]
1-(3-(8-((6-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)piperidin-1-yl)prop-2-en-1-one; [0093]
1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]tr-
iazolo[1,5-a]pyridin-6-yl)piperidin-1-yl)prop-2-en-1-one; [0094]
(S)-1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrid-
in-6-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0095]
(R)-1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrid-
in-6-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0096]
1-(3-(8-((6-morpholinopyridin-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-
-yl)piperidin-1-yl)prop-2-en-1-one; [0097]
1-(3-(8-((6-morpholinopyridin-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-
-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0098]
1-(3-(8-((3-isopropyl-1,2,4-oxadiazol-5-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)piperidin-1-yl)prop-2-en-1-one; [0099]
N-((1R,3S)-3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclohexyl)acrylamide; [0100]
N-((1S,3S)-3-(8-((1-methyl-1H-pyrazol-3-yl)methyl)-[1,2,4]triazolo[1,5-a]-
pyridin-6-yl)cyclohexyl)acrylamide; [0101]
1-(3-(8-((3-methyl-1,2,4-oxadiazol-5-yl)amino)-[1,2,4]triazolo[1,5-a]pyri-
din-6-yl)piperidin-1-yl)prop-2-en-1-one; [0102]
N-((1S,3S)-3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclohexyl)cyanamide; [0103]
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)piperidine-1-carbonitrile; [0104]
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)pyrrolidine-1-carbonitrile; [0105]
1-(4-((6-(1-acryloylpyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)a-
mino)phenyl)-3-methylimidazolidin-2-one; [0106]
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)pyrrolidine-1-carbonitrile; [0107]
N-((1R,3S)-3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclopentyl)acrylamide; [0108] N-((1
S)-3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)cyclopentyl)acrylamide; [0109]
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)piperidine-1-carbonitrile; [0110]
(S)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,-
4]triazolo[1,5-a]pyridin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one;
[0111]
(R)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,-
4]triazolo[1,5-a]pyridin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one;
[0112]
1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-
-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0113]
(S)-1-(3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyraz-
in-6-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0114]
(R)-1-(3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyraz-
in-6-yl)pyrrolidin-1-yl)prop-2-en-1-one; [0115]
(S)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,-
4]triazolo[1,5-a]pyridin-6-yl)piperidin-1-yl)prop-2-en-1-one;
[0116]
1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)imidazo[1,-
2-b]pyridazin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one; or [0117]
(R)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,-
4]triazolo[1,5-a]pyridin-6-yl)piperidin-1-yl)prop-2-en-1-one.
[0118] In an eighth embodiment the invention provides a compound
according to compound according to any of the foregoing embodiments
wherein [0119] R.sup.5 is --R.sup.501-L-R.sup.52 wherein [0120]
R.sup.501 is a bond; [0121] L is -L.sup.1-L.sup.2 wherein L.sup.1
is attached to R.sup.501 wherein [0122] L.sup.1 is optionally
substituted saturated or partially saturated
(C.sub.3-C.sub.7)cycloalkyl and L.sup.2 is a
bond-CH.sub.2N(R.sup.a)C(O)--, or --N(R.sup.a)C(O)--; and [0123]
R.sup.502 is H, CF.sub.3, OH, optionally substituted
(C.sub.1-C.sub.6)alkyl, optionally substituted alkenyl, optionally
substituted alkynyl, CN, or optionally substituted
(C.sub.3-C.sub.6)cycloalkenyl.
[0124] In a ninth embodiment the invention provides a compound
according to compound according to any of the foregoing embodiments
wherein [0125] R.sup.6 is optionally substituted
(C.sub.1-C.sub.6)alkyl, optionally substituted
(C.sub.3-C.sub.12)cycloalkyl, optionally substituted phenyl,
optionally substituted pyrazolyl, optionally substituted
6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazinyl, optionally substituted
4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinyl; or [0126] R.sup.6 is
--R.sup.601--R.sup.602 wherein R.sup.601 is attached to the
--N(H)-- and [0127] R.sup.601 is optionally substituted pyrazolyl,
or optionally substituted pyridinyl; [0128] R.sup.602 is
N(R.sup.a).sub.2, optionally substituted (C.sub.1-C.sub.6)alkyl,
optionally substituted (C.sub.3-C.sub.6)cycloalkyl, optionally
substituted azetidinyl, optionally substituted morpholinyl,
optionally substituted piperidinyl, or optionally substituted
tetrahydropyranyl.
[0129] In a tenth embodiment the invention provides a compound
according to claim 9 wherein R.sup.1 is H.
[0130] In an eleventh embodiment the invention provides a compound
according to compound according to any of the foregoing embodiments
wherein X is N or CR.sup.2 wherein R.sup.2 is H, optionally
substituted (C.sub.1-C.sub.3)alkyl, or CF.sub.3.
[0131] In a twelfth embodiment the invention provides a compound
according to compound according to any of the foregoing embodiments
wherein R.sup.3 is H, deuterium or optionally substituted
(C.sub.1-C.sub.3)alkyl.
[0132] In a thirteenth embodiment the invention provides a
according to compound according to any of the foregoing embodiments
wherein U is CH.
[0133] In a fourteenth embodiment the invention provides a compound
according to compound according to any of the foregoing embodiments
wherein X is N.
[0134] In a fifteenth embodiment the invention provides a compound
according to compound according to any of the foregoing embodiments
wherein the compound is [0135]
4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)-2-methylbutan-2-ol; [0136]
6-cyclohexyl-N-(1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-yl)-[1,2,4]tria-
zolo[1,5-a]pyrazin-8-amine; [0137]
6-cyclohexyl-N-(1-(2,2,6,6-tetramethyltetrahydro-2H-pyran-4-yl)-1H-pyrazo-
l-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amine; [0138]
6-cyclohexyl-N-(1-(oxetan-3-yl)-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]py-
razin-8-amine; [0139]
(1R,4R)-4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H--
pyrazol-1-yl)cyclohexanol; [0140] (1
s,4s)-4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-py-
razol-1-yl)cyclohexanol; [0141]
6-cyclohexyl-N-(1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amine;
[0142]
(6-cyclohexyl-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazo-
lo[1,5-a]pyrazin-8-amine; [0143]
6-cyclopentyl-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5--
a]pyrazin-8-amine; [0144]
6-cyclohexyl-N-(1-isopropyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazi-
n-8-amine; [0145]
6-(4,4-dimethylcyclohexyl)-N-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1-
,5-a]pyrazin-8-amine; [0146]
N-(1-methyl-1H-pyrazol-4-yl)-6-((1R,4R)-4-methylcyclohexyl)-[1,2,4]triazo-
lo[1,5-a]pyrazin-8-amine; [0147]
N-(1-methyl-1H-pyrazol-4-yl)-6-((1S,4S)-4-methylcyclohexyl)-[1,2,4]triazo-
lo[1,5-a]pyrazin-8-amine; [0148]
N-(1-methyl-1H-pyrazol-4-yl)-6-((1R,4R)-4-(trifluoromethyl)cyclohexyl)-[1-
,2,4]triazolo[1,5-a]pyrazin-8-amine; [0149]
N-(6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazo-
lo[5,1-c][1,4]oxazin-2-amine; [0150]
6-cyclohexyl-N-(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)-[-
1,2,4]triazolo[1,5-a]pyridin-8-amine; [0151]
6-cyclopentyl-N-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin--
8-amine; [0152]
6-cyclopentyl-N-(5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazin-2-yl)--
[1,2,4]triazolo[1,5-a]pyridin-8-amine; [0153]
1-(4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyraz-
ol-1-yl)piperidin-1-yl)ethanone; [0154]
4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)-N-methylpiperidine-1-carboxamide; [0155]
(1S,3S)-3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H--
pyrazol-1-yl)cyclohexanol; [0156]
(1R,3R)-3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H--
pyrazol-1-yl)cyclohexanol; [0157]
(1R,3S)-3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H--
pyrazol-1-yl)cyclohexanol; [0158]
(1S,3R)-3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H--
pyrazol-1-yl)cyclohexanol; [0159]
(1R,3R)-3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyra-
zin-6-yl)cyclohexanol; [0160]
1-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)-2-methylpropan-2-ol; [0161]
N-(6-cyclopentyl-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyraz-
olo[5,1-c][1,4]oxazin-2-amine; [0162]
1-(6-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyridin-8-yl)amino)pyridin-3-yl-
)piperidin-4-ol; [0163]
6-cyclohexyl-N-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazo-
lo[1,5-a]pyrazin-8-amine; [0164] (1S,4S)-ethyl
4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)cyclohexanecarboxylate; [0165]
6-cyclopentyl-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5--
a]pyrazin-8-amine; or [0166]
6-cyclohexyl-N-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-
-amine.
[0167] In a sixteenth embodiment, the invention provides a method
of treating a disease comprising administering a therapeutically
effective amount of a compound compound according to any of the
foregoing embodiments to a patient in need thereof.
[0168] In a seventeenth embodiment the invention provides a method
according to the sixteenth embodiment, wherein the disease is
rheumatoid arthritis, juvenile rheumatoid arthritis,
osteoarthritis, Crohn's disease, inflammatory bowel disease,
irritable bowel syndrome, ulcerative colitis, psoriatic arthritis,
psoriasis, ankylosing spondylitis, interstitial cystitis, asthma,
systemic lupus erythematosus, lupus nephritis, B cell chronic
lymphocytic lymphoma, multiple sclerosis, chronic lymphocytic
leukemia, small lymphocytic lymphoma, mantle cell lymphoma, B-cell
non-Hodgkin's lymphoma, activated B-cell like diffuse large B-cell
lymphoma, multiple myelorna, diffuse large B-cell lymphoma,
follicular lymphoma, hairy cell leukemia or Lymphoblastic
lymphoma.
[0169] In an eighteenth embodiment the invention provides a kit
comprising a packaged product comprising components with which to
administer a compound compound according to any of the first
through fifteenth embodiments for treatment of an autoimmune
disorder.
[0170] In a nineteenth embodiment the invention provides a kit
according to the eighteenth embodiment, wherein the packaged
product comprises a compound first through fifteenth embodiments
and instructions for use.
[0171] In a twentieth embodiment the invention provides a
pharmaceutical composition comprising a compound according to any
one of the first to the fifteenth embodiment, and one or more
pharmaceutically acceptable excipients.
DETAILED DESCRIPTION OF THE INVENTION
[0172] Protein kinases are a broad and diverse class, of over 500
enzymes, that include oncogenes, growth factors receptors, signal
transduction intermediates, apoptosis related kinases and cyclin
dependent kinases. They are responsible for the transfer of a
phosphate group to specific tyrosine, serine or threonine amino
acid residues, and are broadly classified as tyrosine and
serine/threonine kinases as a result of their substrate
specificity.
[0173] The protein kinases represent a large family of proteins
that play a central role in the regulation of a wide variety of
cellular processes and maintenance of cellular function. A partial,
non-limiting, list of these kinases include: non-receptor tyrosine
kinases such as the Tec family (BTK, ITK, Tec, ETK/BMX &
RLK/TXK), Janus kinase family (Jak1, Jak2, Jak3 and Tyk2); the
fusion kinases, such as BCR-Abl, focal adhesion kinase (FAK), Fes,
Lck and Syk; receptor tyrosine kinases such as epidermal growth
factor receptor (EGFR), the platelet-derived growth factor receptor
kinase (PDGF-R), the receptor kinase for stem cell factor, c-kit,
the hepatocyte growth factor receptor, c-Met, and the fibroblast
growth factor receptor, FGFR3; and serine/threonine kinases such as
b-RAF, mitogen-activated protein kinases (e.g., MKK6) and
SAPK2.beta.. Aberrant kinase activity has been observed in many
disease states including benign and malignant proliferative
disorders as well as diseases resulting from inappropriate
activation of the immune and nervous systems. The novel compounds
of this invention inhibit the activity of one or more protein
kinases and are, therefore, expected to be useful in the treatment
of kinase-mediated diseases.
[0174] Bruton's tyrosine kinase (BTK) is a non-receptor tyrosine
kinase with a key role in immunoreceptor signaling (BCR,
Fc.epsilon.R, Fc.gamma.R, DAP12, Dectin-1, GPVI, etc.) in a host of
hematopoietic cells including B cells, platelets, mast cells,
basophils, eosinophils, macrophages and neutrophils as well as
osteoclasts involved in bone destruction (for reviews, see Brunner
et al., 2005 Histol. Histopathol., 20:945, Mohamed et al., 2009
Immunol. Rev., 228:58). Mutations in BTK are known to lead to
X-linked agammaglobulinemia (XLA) in humans and X-linked
immunodeficiency (Xid) in mice, which are characterized by limited
B-cell production & reduced antibody titers (Lindvall et al.,
2005 Immunol. Rev., 203:200). The combined action of BTK in
multiple cell types makes it an attractive target for autoimmune
disease. BTK is related with sequence homology to other Tec family
kinases (ITK, Tec, ETK/BMX & RLK/TXK).
[0175] In B-lymphocytes, BTK is required for B-cell development and
for Ca2+ mobilization following of B-cell receptor (BCR) engagement
(Khan et al., 1995 Immunity 3:283; Genevier et al., 1997 Clin. Exp.
Immun., 110:286) where it is believed to downstream of Src family
kinases (such as Lyn), Syk & PI3K. BTK has been shown to be
important for both thymus-dependent and thymus-independent type 2
responses to antigens (Khan et al., Immunity 1995; 3; 283). In mast
cells, studies using BTK mouse knock-outs (Hata et al., 1998 J.
Exp. Med., 187:1235; Schmidt et al., 2009 Eur. J. Immun., 39:3228)
indicate a role for BTK in Fc.epsilon.RI induced signaling,
histamine release & production of cytokines such as TNF, IL-2,
& IL-4. In platelets, BTK is important for signaling through
the glycoprotein VI (GPVI) receptor that responds to collagen and
has been shown to promote platelet aggregation and contribute to
cytokine production from fibroblast-like synoviocytes (Hsu et al.,
2013 Immun. Letters 150:97). In monocytes and macrophages, the
action of BTK in invoked in Fc.gamma.RI induced signaling and may
also have role in Toll-Like Receptor-induced cytokine responses
including TLR2, TLR4, TLR8 & TLR9 (Horwood et al., 2003 J. Exp.
Med., 197:1603; Horwood et al., 2006 J. Immunol., 176:3635; Perez
de Diego et al., 2006 Allerg. Clin. Imm., 117:1462; Doyle et al.,
2007 J. Biol. Chem., 282:36959, Hasan et al., 2007 Immunology,
123:239; Sochorava et al., 2007 Blood, 109:2553; Lee et al., 2008,
J. Biol. Chem., 283:11189).
[0176] Therefore, inhibition of BTK is expected to intervene at
several critical junctions of the inflammatory reactions resulting
in an effective suppression of autoimmune response. As such
diseases involving B-cell receptor activation, antibody-Fc receptor
interactions & GPVI receptor signaling may be modulated by
treatment with BTK inhibitors. BTK inhibition is likely to act on
both the initiation of autoimmune disease by blocking BCR signaling
and the effector phase by abrogation of FcR signaling on
macrophages, neutrophils, basophils, and mast cells. Furthermore,
blocking BTK would provide additional benefit via inhibition of
osteoclast maturation and therefore attenuate the bone erosions
& overall joint destruction associated with rheumatoid
arthritis. Inhibiting BTK may be useful in treating a host of
inflammatory and allergic diseases--for example (but not limited
to), rheumatoid arthritis (RA), systemic lupus erythematosus (SLE),
multiple sclerosis (MS) and type I hypersensitivity reactions such
as allergic rhinitis, allergic conjunctivitis, atopic dermatitis,
allergic asthma and systemic anaphylaxis. For a review on targeting
BTK as a treatment for inflammatory disorders and autoimmunity as
well as leukemias and lymphomas, see Uckun & Qazi 2010 Expert
Opin Ther Pat 20:1457. Because BTK is highly expressed in cancers
of the hematopoietic system & BTK-dependent signaling in
believed to be disregulated there, BTK inhibitors are expected to
be useful treatments for B-cell lymphomas/leukemias & other
oncologic disease for example (but not limited to) acute
lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL),
non-Hodgkin's lymphoma (NHL), small lymphocytic lymphoma (SLL), and
acute myeloid leukemia (for review, see Buggy & Elias 2012 Int
Rev Immunol. 31:119). Taken together, BTK inhibitors provide a
strong method to treat a host of inflammatory diseases and
immunological disorders as well as hematologic cancers.
[0177] All kinases bind a common molecule, ATP, and therefore have
structurally similar binding pockets. Therefore, one of the
challenges for any kinase inhibitor is that they are prone to
inhibit more than one kinase due to the homology of the binding
pocket. For example, staurosporine, a well characterized
promiscuous kinase inhibitor, has been shown to inhibit at least
253 with a k.sub.d of <3 .mu.M kinases from the human kinome
(see Nature Biotechnology, 208, 26, p. 127). Additionally, several
marketed kinase inhibitors are known to inhibit more than one
intended kinase, for example Imatinib (Gleevec.RTM.) targets ABL,
ARG, PDGFR-.alpha./.beta. and c-KIT kinases, sorafenib
(Nexavar.RTM.) targets B-RAF, VEGFRs, PDGFR-.alpha./.beta., FLT3
and c-KIT and sunitinib (Sutent.RTM.) targets VEGFR, PDGFR, CSF-1R,
FLT3 and c-KIT (Nature Reviews Drug Discovery 2011, 10, 111).
[0178] Inhibition of certain kinases in the human kinome are known
to have undesired effects when used as pharmaceutical treatment.
For instance, a number of kinase targets have been implicated in
playing a role in the cardiotoxicity profiles for kinase inhibitors
that are currently on the market. These kinases can include, but
not limited to, VEGFR2, PI3K, AKT, PDGFR-.alpha./.beta., AMPK,
GSK3, ERKs, CDK2, Aurora, PLK, JNK, CAMKII<PDK1, mTOR, LKB1,
CAMKK.beta., MEK1/2, PKA, PKC.alpha., RAF1, B-RAF, EGFR, ERBB2,
c-Kit, ABL, ARG, JAK2, FAK, DMPK, LTK, ROCK, LKB1, LDB3, PIM, GRK2,
GRK5, ASK1, and PTEN (see Nature Reviews Drug Discovery 2011,
10:111). One example from a marketed kinase inhibitor is that in
clinical trials with sunitinib, patients were found to be at
increased risk for hypertension (see The Lancet 2006, 368:1329; and
J. Clin. Oncol. 2009, 27:3584). Subsequent research on the
mechanism for the increased hypertension suggest that while PDGFR
and VEGFR may be playing a role, off-target kinase inhibition, such
as AMPK, may also be contributing to sunitinib's increased risk for
hypertension (Curr. Hypertens. Rep. 2011, 13:436). Additionally,
there is a patent application, US 2011/0212461, that has been filed
that is a method for the prediction of cardiotoxicity based on the
activity versus a list of kinases including KIT, FYN, PDGFR beta,
FGR, LCK, Ephrin Receptor B2, FRK, ABL1, PDGFR1 alpha, HCK, ABL2,
LYN, ZAK, YES1, MAP4K4, PKN1, BRAF, DDR2, MAP4K5 and STK24.
Therefore, identification of kinase inhibitors with a selective
profile Btk or CSF-1R kinase are desirable. The compounds of this
invention are selective for the inhibition of Btk or CSF-1R over
other kinases.
[0179] Many of the kinases, whether a receptor or non-receptor
tyrosine kinase or a S/T kinase have been found to be involved in
cellular signaling pathways involved in numerous pathogenic
conditions, including immunomodulation, inflammation, or
proliferative disorders such as cancer.
[0180] Many autoimmune diseases and disease associated with chronic
inflammation, as well as acute responses, have been linked to
excessive or unregulated production or activity of one or more
cytokines.
[0181] The compounds of the invention are also useful in the
treatment of rheumatoid arthritis, asthma, allergic asthma,
osteoarthritis, juvenile arthritis, ankylosing spondylitis, an
ocular condition, interstitial cystitis, a cancer, a solid tumor, a
sarcoma, fibrosarcoma, osteoma, melanoma, retinoblastoma, a
rhabdomyosarcoma, glioblastoma, neuroblastoma, teratocarcinoma,
hypersensitivity reactions, hyperkinetic movement disorders,
hypersensitivity pneumonitis, hypertension, hypokinetic movement
disorders, aordic and peripheral aneuryisms,
hypothalamic-pituitary-adrenal axis evaluation, aortic dissection,
arterial hypertension, arteriosclerosis, arteriovenous fistula,
ataxia, spinocerebellar degenerations, streptococcal myositis,
structural lesions of the cerebellum, Subacute sclerosing
panencephalitis, Syncope, syphilis of the cardiovascular system,
systemic anaphalaxis, systemic inflammatory response syndrome,
systemic onset juvenile rheumatoid arthritis, T-cell or FAB ALL,
Telangiectasia, thromboangitis obliterans, transplants,
trauma/hemorrhage, type III hypersensitivity reactions, type IV
hypersensitivity, unstable angina, uremia, urosepsis, urticaria,
valvular heart diseases, varicose veins, vasculitis, venous
diseases, venous thrombosis, ventricular fibrillation, viral and
fungal infections, vital encephalitis/aseptic meningitis,
vital-associated hemaphagocytic syndrome, Wernicke-Korsakoff
syndrome, Wilson's disease, xenograft rejection of any organ or
tissue, heart transplant rejection, hemachromatosis, hemodialysis,
hemolytic urenic syndrome/thrombolytic thrombocytopenic purpura,
hemorrhage, idiopathic pulmonary fibrosis, antibody mediated
cytotoxicity, Asthenia, infantile spinal muscular atrophy,
inflammation of the aorta, influenza A, ionizing radiation
exposure, iridocyclitis/uveitis/optic neuritis, juvenile spinal
muscular atrophy, lymphoma, myeloma, leukaemia, malignant ascites,
hematopoietic cancers, a diabetic condition such as
insulin-dependent diabetes mellitus glaucoma, diabetic retinopathy
or microangiopathy, sickle cell anaemia, chronic inflammation,
glomerulonephritis, graft rejection, Lyme disease, von Hippel
Lindau disease, pemphigoid, Paget's disease, fibrosis, sarcoidosis,
cirrhosis, thyroiditis, hyperviscosity syndrome, Osler-Weber-Rendu
disease, chronic occlusive pulmonary disease, asthma or edema
following burns, trauma, radiation, stroke, hypoxia, ischemia,
ovarian hyperstimulation syndrome, post perfusion syndrome, post
pump syndrome, post-MI cardiotomy syndrome, preeclampsia,
menometrorrhagia, endometriosis, pulmonary hypertension, infantile
hemangioma, or infection by Herpes simplex, Herpes Zoster, human
immunodeficiency virus, parapoxvirus, protozoa or toxoplasmosis,
progressive supranucleo palsy, primary pulmonary hypertension,
radiation therapy, Raynaud's phenomenon, Raynaud's disease,
Refsum's disease, regular narrow QRS tachycardia, renovascular
hypertension, restrictive cardiomyopathy, sarcoma, senile chorea,
senile dementia of Lewy body type, shock, skin allograft, skin
changes syndrome, ocular or macular edema, ocular neovascular
disease, scleritis, radial keratotomy, uveitis, vitritis, myopia,
optic pits, chronic retinal detachment, post-laser treatment
complications, conjunctivitis, Stargardt's disease, Eales disease,
retinopathy, macular degeneration, restenosis, ischemia/reperfusion
injury, ischemic stroke, vascular occlusion, carotid obstructive
disease, ulcerative colitis, inflammatory bowel disease, irritable
bowel syndrome, diabetes, diabetes mellitus, insulin dependent
diabetes mellitus, allergic diseases, dermatitis scleroderma, graft
versus host disease, organ transplant rejection (including but not
limited to bone marrow and solid organ rejection), acute or chronic
immune disease associated with organ transplantation, sarcoidosis,
disseminated intravascular coagulation, Kawasaki's disease,
nephrotic syndrome, chronic fatigue syndrome, Wegener's
granulomatosis, Henoch-Schoenlein purpurea, microscopic vasculitis
of the kidneys, chronic active hepatitis, septic shock, toxic shock
syndrome, sepsis syndrome, cachexia, infectious diseases, parasitic
diseases, acquired immunodeficiency syndrome, acute transverse
myelitis, Huntington's chorea, stroke, primary biliary cirrhosis,
hemolytic anemia, malignancies, Addison's disease, idiopathic
Addison's disease, sporadic, polyglandular deficiency type I and
polyglandular deficiency type TI, Schmidt's syndrome, adult (acute)
respiratory distress syndrome, alopecia, alopecia areata,
seronegative arthopathy, arthropathy, Reiter's disease, psoriatic
arthropathy, ulcerative colitic arthropathy, enteropathic
synovitis, chlamydia, yersinia and salmonella associated
arthropathy, atheromatous disease/arteriosclerosis, atopic allergy,
autoimmune bullous disease, pemphigus vulgaris, pemphigus
foliaceus, pemphigoid, linear IgA disease, autoimmune haemolytic
anaemia, Coombs positive haemolytic anaemia, acquired pernicious
anaemia, juvenile pernicious anaemia, peripheral vascular
disorders, peritonitis, pernicious anemia, myalgic
encephalitis/Royal Free Disease, chronic mucocutaneous candidiasis,
giant cell arteritis, primary sclerosing hepatitis, cryptogenic
autoimmune hepatitis, Acquired Immunodeficiency Disease Syndrome,
Acquired Immunodeficiency Related Diseases, Hepatitis A, Hepatitis
B, Hepatitis C, His bundle arrythmias, HIV infection/HIV
neuropathy, common varied immunodeficiency (common variable
hypogammaglobulinaemia), dilated cardiomyopathy, female
infertility, ovarian failure, premature ovarian failure, fibrotic
lung disease, chronic wound healing, cryptogenic fibrosing
alveolitis, post-inflammatory interstitial lung disease,
interstitial pneumonitis, pneumocystis carinii pneumonia,
pneumonia, connective tissue disease associated interstitial lung
disease, mixed connective tissue disease, associated lung disease,
systemic sclerosis associated interstitial lung disease, rheumatoid
arthritis associated interstitial lung disease, systemic lupus
erythematosus associated lung disease, dermatomyositis/polymyositis
associated lung disease, Sjogren's disease associated lung disease,
ankylosing spondylitis associated lung disease, vasculitic diffuse
lung disease, hemosiderosis associated lung disease, drug-induced
interstitial lung disease, radiation fibrosis, bronchiolitis
obliterans, chronic eosinophilic pneumonia, lymphocytic
infiltrative lung disease, postinfectious interstitial lung
disease, gouty arthritis, autoimmune hepatitis, type-1 autoimmune
hepatitis (classical autoimmune or lupoid hepatitis), type-2
autoimmune hepatitis (anti-LKM antibody hepatitis), autoimmune
mediated hypoglycaemia, type B insulin resistance with acanthosis
nigricans, hypoparathyroidism, acute immune disease associated with
organ transplantation, chronic immune disease associated with organ
transplantation, osteoarthritis, primary sclerosing cholangitis,
psoriasis type 1, psoriasis type 2, idiopathic leucopaenia,
autoimmune neutropaenia, renal disease NOS, glomerulonephritides,
microscopic vasculitis of the kidneys, Lyme disease, discoid lupus
erythematosus, male infertility idiopathic or NOS, sperm
autoimmunity, multiple sclerosis (all subtypes), sympathetic
ophthalmia, pulmonary hypertension secondary to connective tissue
disease, acute and chronic pain (different forms of pain),
Goodpasture's syndrome, pulmonary manifestation of polyarteritis
nodosa, acute rheumatic fever, rheumatoid spondylitis, Still's
disease, systemic sclerosis, Sjogren's syndrome, Takayasu's
disease/arteritis, autoimmune thrombocytopenia, toxicity,
transplants, and diseases involving inappropriate vascularization
for example diabetic retinopathy, retinopathy of prematurity,
choroidal neovascularization due to age-related macular
degeneration, and infantile hemangiomas in human beings. In
addition, such compounds may be useful in the treatment of
disorders such as ascites, effusions, and exudates, including for
example macular edema, cerebral edema, acute lung injury, adult
respiratory distress syndrome (ARDS), proliferative disorders such
as restenosis, fibrotic disorders such as hepatic cirrhosis and
atherosclerosis, mesangial cell proliferative disorders such as
diabetic nephropathy, malignant nephrosclerosis, thrombotic
microangiopathy syndromes, and glomerulopathies, myocardial
angiogenesis, coronary and cerebral collaterals, ischemic limb
angiogenesis, ischemia/reperfusion injury, peptic ulcer
Helicobacter related diseases, virally-induced angiogenic
disorders, preeclampsia, menometrorrhagia, cat scratch fever,
rubeosis, neovascular glaucoma and retinopathies such as those
associated with diabetic retinopathy, retinopathy of prematurity,
or age-related macular degeneration. In addition, these compounds
can be used as active agents against hyperproliferative disorders
such as thyroid hyperplasia (especially Grave's disease), and cysts
(such as hypervascularity of ovarian stroma characteristic of
polycystic ovarian syndrome (Stein-Leventhal syndrome) and
polycystic kidney disease since such diseases require a
proliferation of blood vessel cells for growth and/or
metastasis.
[0182] In yet other embodiments, the compounds described herein can
be used to treat a cancer, e.g., B-cell proliferative disorders,
which include, but are not limited to diffuse large B cell
lymphoma, follicular lymphoma, chronic lymphocytic lymphoma,
chronic lymphocytic leukemia, B-cell prolymphocytic leukemia,
lymphoplamacytic lymphoma/Waldenstrom macroglobulinemia, splenic
marginal zone lymphoma, plasma cell myeloma, plasmacytoma,
extranodal marginal zone B cell lymphoma, nodal marginal zone B
cell lymphoma, mantle cell lymphoma, mediastinal (thymic) large B
cell lymphoma, intravascular large B cell lymphoma, primary
effusion lymphoma, Burkitt's lymphoma/leukemia, lymphomatoid
granulomatosis, pancreatic cancer, solid or hematological tumors, a
benign or malignant tumor, carcinoma of the brain, kidney (e.g.,
renal cell carcinoma (RCC)), squamous cell carcinoma, salivary
gland carcinoma, liver, adrenal gland, bladder, breast, stomach,
gastric tumors, ovaries, colon, rectum, prostate, pancreas, lung,
vagina, endometrium, cervix, testis, genitourinary tract,
esophagus, larynx, skin, bone or thyroid, sarcoma, glioblastomas,
neuroblastomas, multiple myeloma or gastrointestinal cancer,
especially colon carcinoma or colorectal adenoma or a tumor of the
neck and head, an epidermal hyperproliferation, psoriasis, prostate
hyperplasia, a neoplasia, a neoplasia of epithelial character,
adenoma, adenocarcinoma, keratoacanthoma, epidermoid carcinoma,
large cell carcinoma, non-small-cell lung carcinoma, lymphomas,
(including, for example, non-Hodgkin's Lymphoma (NHL) and Hodgkin's
lymphoma (also termed Hodgkin's or Hodgkin's disease)), a mammary
carcinoma, follicular carcinoma, undifferentiated carcinoma,
papillary carcinoma, seminoma, melanoma, or a leukemia.
[0183] In yet other embodiments, the compounds described herein can
be used to treat Behcet's disease, osteoporosis, bone cancer, and
bone metastasis, systemic sclerosis, contact dermatitis and other
eczematous dermatitis, seborrhoeic dermatitis, lichen planus,
epidermolysis bullosa, angiodermas, vasculitides, cutaneous
eosinophilias, or vernal conjunctivitis.
[0184] In yet other embodiments, the compounds described herein can
be used to treat those conditions characterized by inflammation of
the nasal mucus membrane, including acute rhinitis, allergic,
atrophic thinitis and chronic rhinitis including rhinitis caseosa,
hypertrophic rhinitis, rhinitis purulenta, rhinitis sicca and
rhinitis medicamentosa; membranous rhinitis including croupous,
fibrinous and pseudomembranous rhinitis and scrofulous rhinitis,
seasonal rhinitis including rhinitis nervosa (hay fever) and
vasomotor rhinitis, sarcoidosis, farmer's lung and related
diseases, fibroid lung, and idiopathic interstitial pneumonia.
[0185] Compounds of Formula (I) of the invention can be used alone
or in combination with an additional agent, e.g., a therapeutic
agent, said additional agent being selected by the skilled artisan
for its intended purpose. For example, the additional agent can be
a therapeutic agent art-recognized as being useful to treat the
disease or condition being treated by the compound of the present
invention. The additional agent also can be an agent that imparts a
beneficial attribute to the therapeutic composition e.g., an agent
that affects the viscosity of the composition.
[0186] It should further be understood that the combinations which
are to be included within this invention are those combinations
useful for their intended purpose. The agents set forth below are
illustrative for purposes and not intended to be limited. The
combinations, which are part of this invention, can be the
compounds of the present invention and at least one additional
agent selected from the lists below. The combination can also
include more than one additional agent, e.g., two or three
additional agents if the combination is such that the formed
composition can perform its intended function.
[0187] Preferred combinations are non-steroidal anti-inflammatory
drug(s) also referred to as NSAIDS which include drugs like
ibuprofen. Other preferred combinations are corticosteroids
including prednisolone; the well known side-effects of steroid use
can be reduced or even eliminated by tapering the steroid dose
required when treating patients in combination with the compounds
of this invention. Non-limiting examples of therapeutic agents for
rheumatoid arthritis with which a compound of Formula (I) of the
invention can be combined include the following: cytokine
suppressive anti-inflammatory drug(s) (CSAIDs); antibodies to or
antagonists of other human cytokines or growth factors, for
example, TNF, LT, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8,
IL-12, IL-15, IL-16, IL-21, IL-23, interferons, EMAP-II, GM-CSF,
FGF, MMP-13 and PDGF. Compounds of the invention can be combined
with antibodies to cell surface molecules such as CD2, CD3, CD4,
CD8, CD25, CD28, CD30, CD40, CD45, CD69, CD80 (B7.1), CD86 (B7.2),
CD90, CTLA or their ligands including CD154 (gp39 or CD40L).
[0188] Preferred combinations of therapeutic agents may interfere
at different points in the autoimmune and subsequent inflammatory
cascade; preferred examples include TNF antagonists like chimeric,
humanized or human TNF antibodies, D2E7 (U.S. Pat. No. 6,090,382,
HUMIRA.TM.), CA2 (REMICADE.TM.), SIMPONI.TM. (golimumab),
CIMZIA.TM., ACTEMRA.TM., CDP 571, and soluble p55 or p75 TNF
receptors, derivatives, thereof, (p75TNFR1gG (ENBREL.TM.) or
p55TNFR1gG (Lenercept), and also TNF.alpha. converting enzyme
(TACE) inhibitors; similarly IL-1 inhibitors
(Interleukin-1-converting enzyme inhibitors, IL-1RA etc.) may be
effective for the same reason. Other preferred combinations include
Interleukin 11. Yet other preferred combinations are the other key
players of the autoimmune response which may act parallel to,
dependent on or in concert with IL-18 function; especially
preferred are IL-12 antagonists including IL-12 antibodies or
soluble IL-12 receptors, or IL-12 binding proteins. It has been
shown that IL-12 and IL-18 have overlapping but distinct functions
and a combination of antagonists to both may be most effective. Yet
another preferred combination is non-depleting anti-CD4 inhibitors.
Yet other preferred combinations include antagonists of the
co-stimulatory pathway CD80 (B7.1) or CD86 (B7.2) including
antibodies, soluble receptors or antagonistic ligands.
[0189] A compound of Formula (I) of the invention may also be
combined with agents, such as methotrexate, 6-mercaptopurine,
azathioprine sulphasalazine, mesalazine, olsalazine
chloroquinone/hydroxychloroquine, penacillamine, aurothiomalate
(intramuscular and oral), azathioprine, colchicine, corticosteroids
(oral, inhaled and local injection), beta-2 adrenoreceptor agonists
(salbutamol, terbutaline, salmeteral), xanthines (theophylline,
aminophylline), cromoglycate, nedocromil, ketotifen, ipratropium
and oxitropium, cyclosporin, FK506, rapamycin, mycophenolate
mofetil, leflunomide, NSAIDs, for example, ibuprofen,
corticosteroids such as prednisolone, phosphodiesterase inhibitors,
adensosine agonists, antithrombotic agents, complement inhibitors,
adrenergic agents, agents which interfere with signalling by
proinflammatory cytokines such as TNF.alpha. or IL-1 (e.g., NIK,
IKK, JAK1, JAK2, JAK3, p38 or MAP kinase inhibitors), IL-13
converting enzyme inhibitors, T-cell signalling inhibitors such as
kinase inhibitors, metalloproteinase inhibitors, sulfasalazine,
6-mercaptopurines, angiotensin converting enzyme inhibitors,
soluble cytokine receptors and derivatives thereof (e.g. soluble
p55 or p75 TNF receptors and the derivatives p75TNFRIgG
(Enbrel.TM.) and p55TNFRIgG (Lenercept), sIL-1RI, sIL-1RII,
sIL-6R), antiinflammatory cytokines (e.g. IL-4, IL-10, IL-11, IL-13
and TGF.beta.), cclccoxib, folic acid, hydroxychloroquine sulfate,
rofecoxib, etanercept, infliximab, naproxen, valdecoxib,
sulfasalazine, methylprednisolone, meloxicam, methylprednisolone
acetate, gold sodium thiomalate, aspirin, triamcinolone acetonide,
propoxyphene napsylate/apap, folate, nabumetone, diclofenac,
piroxicam, etodolac, diclofenac sodium, oxaprozin, oxycodone HCl,
hydrocodone bitartrate/apap, diclofenac sodium/misoprostol,
fentanyl, anakinra, tramadol HCl, salsalate, sulindac,
cyanocobalamin/fa/pyridoxine, acetaminophen, alendronate sodium,
prednisolone, morphine sulfate, lidocaine hydrochloride,
indomethacin, glucosamine sulf/chondroitin, amitriptyline HCl,
sulfadiazine, oxycodone HCl/acetaminophen, olopatadine HCl
misoprostol, naproxen sodium, omeprazole, cyclophosphamide,
rituximab, tofacitinib, IL-1 TRAP, MRA, CTLA4-IG, IL-18 BP,
anti-IL-12, Anti-IL15, BIRB-796, SCIO-469, VX-702, AMG-548, VX-740,
Roflumilast, IC-485, CDC-801, S1P1 agonists (such as FTY720), PKC
family inhibitors (such as Ruboxistaurin or AEB-071) and Mesopram.
Preferred combinations include methotrexate or leflunomide and in
moderate or severe rheumatoid arthritis cases, cyclosporine and
anti-TNF antibodies as noted above.
[0190] Non-limiting examples of therapeutic agents for inflammatory
bowel disease with which a compound of Formula (I) of the invention
can be combined include the following: budenoside; epidermal growth
factor; corticosteroids; cyclosporin, sulfasalazine;
aminosalicylates; 6-mercaptopurine; azathioprine; metronidazole;
lipoxygenase inhibitors; mesalamine; olsalazine; balsalazide;
antioxidants; thromboxane inhibitors; IL-1 receptor antagonists;
anti-IL-13 monoclonal antibodies; anti-IL-6 monoclonal antibodies;
growth factors; elastase inhibitors; pyridinyl-imidazole compounds;
antibodies to or antagonists of other human cytokines or growth
factors, for example, TNF, LT, IL-1, IL-2, IL-6, IL-7, IL-8, IL-12,
IL-15, IL-16, IL-23, EMAP-II, GM-CSF, FGF, and PDGF; cell surface
molecules such as CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45,
CD69, CD90 or their ligands; methotrexate; cyclosporine; FK506;
rapamycin; mycophenolate mofetil; leflunomide; NSAIDs, for example,
ibuprofen; corticosteroids such as prednisolone; phosphodiesterase
inhibitors; adenosine agonists; antithrombotic agents; complement
inhibitors; adrenergic agents; agents which interfere with
signalling by proinflanmuatory cytokines such as TNF.alpha. or IL-1
(e.g. NIK, IKK, p38 or MAP kinase inhibitors); IL-1f converting
enzyme inhibitors; TNF.alpha. converting enzyme inhibitors; T-cell
signalling inhibitors such as kinase inhibitors; metalloproteinase
inhibitors; sulfasalazine; azathioprine; 6-mercaptopurines;
angiotensin converting enzyme inhibitors; soluble cytokine
receptors and derivatives thereof (e.g. soluble p55 or p75 TNF
receptors, sIL-1RI, sIL-1RII, sIL-6R) and antiinflammatory
cytokines (e.g. IL-4, IL-10, IL-11, IL-13 and TGF.beta.). Preferred
examples of therapeutic agents for Crohn's disease with which a
compound of Formula (I) can be combined include the following: TNF
antagonists, for example, anti-TNF antibodies, D2E7 (U.S. Pat. No.
6,090,382, HUMIRA.TM.), CA2 (REMICADE.TM.), CDP 571, TNFR-Ig
constructs, (p75TNFRIgG (ENBREL.TM.) and p55TNFRIgG (LENERCEPT.TM.)
inhibitors and PDE4 inhibitors. A compound of Formula (I) can be
combined with corticosteroids, for example, budenoside and
dexamethasone; sulfasalazine, 5-aminosalicylic acid; olsalazine;
and agents which interfere with synthesis or action of
proinflammatory cytokines such as IL-1, for example, IL-1.beta.
converting enzyme inhibitors and IL-1ra; T cell signaling
inhibitors, for example, tyrosine kinase inhibitors;
6-mercaptopurine; IL-11; mesalamine; prednisone; azathioprine;
mercaptopurine; infliximab; methylprednisolone sodium succinate;
diphenoxylate/atrop sulfate; loperamide hydrochloride;
methotrexate; orneprazole; folate; ciprofloxacin/dextrose-water;
hydrocodone bitartrate/apap; tetracycline hydrochloride;
fluocinonide; metronidazole; thimerosal/boric acid;
cholestyramine/sucrose; ciprofloxacin hydrochloride; hyoscyamine
sulfate; meperidine hydrochloride; midazolam hydrochloride;
oxycodone HCl/acetaminophen; promethazine hydrochloride; sodium
phosphate; sulfamethoxazole/trimethoprim; celecoxib; polycarbophil;
propoxyphene napsylate; hydrocortisone; multivitamins; balsalazide
disodium; codeine phosphate/apap; colesevelam HCl; cyanocobalamin;
folic acid; levofloxacin; methylprednisolone; natalizumab and
interferon-gamma.
[0191] Non-limiting examples of therapeutic agents for multiple
sclerosis with which a compound of Formula (I) can be combined
include the following: corticosteroids; prednisolone;
methylprednisolone; azathioprine; cyclophosphamide; cyclosporine;
methotrexate; 4-aminopyridine; tizanidine; interferon-.beta.1a
(AVONEX.RTM.; Biogen); interferon-.beta.1b (BETASERON.RTM.;
Chiron/Berlex); interferon .alpha.-n3) (Interferon
Sciences/Fujimoto), interferon-.alpha. (Alfa Wassermann/J&J),
interferon .beta.1A-IF (Serono/Inhale Therapeutics), Peginterferon
.alpha. 2b (Enzon/Schering-Plough), Copolymer 1 (Cop-1;
COPAXONE.RTM.; Teva Pharmaceutical Industries, Inc.); hyperbaric
oxygen; intravenous immunoglobulin; cladribine; antibodies to or
antagonists of other human cytokines or growth factors and their
receptors, for example, TNF, LT, IL-1, IL-2, IL-6, IL-7, IL-8,
IL-12, IL-23, IL-15, IL-16, EMAP-II, GM-CSF, FGF, and PDGF. A
compound of Formula (I) can be combined with antibodies to cell
surface molecules such as CD2, CD3, CD4, CD8, CD19, CD20, CD25,
CD28, CD30, CD40, CD45, CD69, CD80, CD86, CD90 or their ligands. A
compound of Formula (I) may also be combined with agents such as
methotrexate, cyclosporine, FK506, rapamycin, mycophenolate
mofetil, leflunomide, an S1P1 agonist, NSAIDs, for example,
ibuprofen, corticosteroids such as prednisolone, phosphodiesterase
inhibitors, adensosine agonists, antithrombotic agents, complement
inhibitors, adrenergic agents, agents which interfere with
signalling by proinflammatory cytokines such as TNF.alpha. or IL-1
(e.g., NIK, IKK, p38 or MAP kinase inhibitors), IL-1.beta.
converting enzyme inhibitors, TACE inhibitors, T-cell signaling
inhibitors such as kinase inhibitors, metalloproteinase inhibitors,
sulfasalazine, azathioprine, 6-mercaptopurines, angiotensin
converting enzyme inhibitors, soluble cytokine receptors and
derivatives thereof (e.g. soluble p55 or p75 TNF receptors,
sIL-1RI, sIL-1RII, sIL-6R) and antiinflammatory cytokines (e.g.
IL-4, IL-10, IL-13 and TGF.beta.).
[0192] Preferred examples of therapeutic agents for multiple
sclerosis in which a compound of Formula (I) can be combined to
include interferon-.beta., for example, IFN.beta.1a and
IFN.beta.1b; copaxone, corticosteroids, caspase inhibitors, for
example inhibitors of caspase-1, IL-1 inhibitors, TNF inhibitors,
and antibodies to CD40 ligand and CD80.
[0193] A compound of Formula (I) may also be combined with agents,
such as alemtuzumab, dronabinol, daclizumab, mitoxantrone,
xaliproden hydrochloride, fampridine, glatiramer acetate,
natalizumab, sinnabidol, .alpha.-immunokine NNSO3, ABR-215062,
AnergiX.MS, chemokine receptor antagonists, BBR-2778, calagualine,
CPI-1189, LEM (liposome encapsulated mitoxantrone), THC.CBD
(cannabinoid agonist), MBP-8298, mesopram (PDE4 inhibitor),
MNA-715, anti-IL-6 receptor antibody, neurovax, pirfenidone
allotrap 1258 (RDP-1258), sTNF-R1, talampanel, teriflunomide,
TGF-beta2, tiplimotide, VLA-4 antagonists (for example, TR-14035,
VLA4 Ultrahaler, Antegran-ELAN/Biogen), interferon gamma
antagonists and IL-4 agonists.
[0194] Non-limiting examples of therapeutic agents for ankylosing
spondylitis with which a compound of Formula (I) can be combined
include the following: ibuprofen, diclofenac, misoprostol,
naproxen, meloxicam, indomethacin, diclofenac, celecoxib,
rofecoxib, sulfasalazine, methotrexate, azathioprine, minocyclin,
prednisone, and anti-TNF antibodies, D2E7 (U.S. Pat. No. 6,090,382;
HUMIRA.TM.), CA2 (REMICADE.TM.), CDP 571, TNFR-Ig constructs,
(p75TNFRIgG (ENBREL.TM.) and p55TNFRIgG (LENERCEPT.TM.).
[0195] Non-limiting examples of therapeutic agents for asthma with
which a compound of Formula (I) can be combined include the
following: albuterol, salmeterol/fluticasone, montelukast sodium,
fluticasone propionate, budesonide, prednisone, salmeterol
xinafoate, levalbuterol HCl, albuterol sulfate/ipratropium,
prednisolone sodium phosphate, triamcinolone acetonide,
beclomethasone dipropionate, ipratropium bromide, azithromycin,
pirbuterol acetate, prednisolone, theophylline anhydrous,
methylprednisolone sodium succinate, clarithromycin, zafirlukast,
formoterol fumarate, influenza virus vaccine, amoxicillin
trihydrate, flunisolide, allergy injection, cromolyn sodium,
fexofenadine hydrochloride, flunisolide/menthol,
amoxicillin/clavulanate, levofloxacin, inhaler assist device,
guaifenesin, dexamethasone sodium phosphate, moxifloxacin HCl,
doxycycline hyclate, guaifenesin/d-methorphan,
p-ephedrine/cod/chlorphenir, gatifloxacin, cetirizine
hydrochloride, mometasone furoate, salmeterol xinafoate,
benzonatate, cephalexin, pe/hydrocodone/chlorphenir, cetirizine
HCl/pseudoephed, phenylephrine/cod/promethazine,
codeine/promethazine, cefprozil, dexamethasone,
guaifenesin/pseudoephedrine, chlorpheniramine/hydrocodone,
nedocromil sodium, terbutaline sulfate, epinephrine,
methylprednisolone, anti-IL-13 antibody, and metaproterenol
sulfate.
[0196] Non-limiting examples of therapeutic agents for COPD with
which a compound of Formula (I) can be combined include the
following: albuterol sulfate/ipratropium, ipratropium bromide,
salmeterol/fluticasone, albuterol, salmeterol xinafoate,
fluticasone propionate, prednisone, theophylline anhydrous,
methylprednisolone sodium succinate, montelukast sodium,
budesonide, formoterol fumarate, triamcinolone acetonide,
levofloxacin, guaifenesin, azithromycin, beclomethasone
dipropionate, levalbuterol HCl, flunisolide, ceftriaxone sodium,
amoxicillin trihydrate, gatifloxacin, zafirlukast,
amoxicillin/clavulanate, flunisolide/menthol,
chlorpheniramine/hydrocodone, metaproterenol sulfate,
methylprednisolone, mometasone furoate,
p-ephedrine/cod/chlorphenir, pirbuterol acetate,
p-ephedrine/loratadine, terbutaline sulfate, tiotropium bromide,
(R,R)-formoterol, TgAAT, cilomilast and roflumilast.
[0197] Non-limiting examples of therapeutic agents for HCV with
which a compound of Formula (I) (can be combined include the
following: Interferon-alpha-2.alpha., Interferon-alpha-2.beta.,
Interferon-alpha con1, Interferon-alpha-n1, pegylated
interferon-alpha-2.alpha., pegylated interferon-alpha-2.beta.,
ribavirin, peginterferon alfa-2b+ribavirin, ursodeoxycholic acid,
glycyrrhizic acid, thymalfasin, Maxamine, VX-497 and any compounds
that are used to treat HCV through intervention with the following
targets: HCV polymerase, HCV protease, HCV helicase, and HCV IRES
(internal ribosome entry site).
[0198] Non-limiting examples of therapeutic agents for Idiopathic
Pulmonary Fibrosis with which a compound of Formula (I) (can be
combined include the following: prednisone, azathioprine,
albuterol, colchicine, albuterol sulfate, digoxin, gamma
interferon, methylprednisolone sodium succinate, lorazepam,
furosemide, lisinopril, nitroglycerin, spironolactone,
cyclophosphamide, ipratropium bromide, actinomycin d, alteplase,
fluticasone propionate, levofloxacin, metaproterenol sulfate,
morphine sulfate, oxycodone HCl, potassium chloride, triamcinolone
acetonide, tacrolimus anhydrous, calcium, interferon-alpha,
methotrexate, mycophenolate mofetil and
interferon-gamma-1.beta..
[0199] Non-limiting examples of therapeutic agents for myocardial
infarction with which a compound of Formula (I) can be combined
include the following: aspirin, nitroglycerin, metoprolol tartrate,
enoxaparin sodium, heparin sodium, clopidogrel bisulfate,
carvedilol, atenolol, morphine sulfate, metoprolol succinate,
warfarin sodium, lisinopril, isosorbide mononitrate, digoxin,
furosemide, simvastatin, ramipril, tenecteplase, enalapril maleate,
torsemide, retavase, losartan potassium, quinapril
hydrochloride/magnesium carbonate, bumetanide, alteplase,
enalaprilat, amiodarone hydrochloride, tirofiban HCl m-hydrate,
diltiazem hydrochloride, captopril, irbesartan, valsartan,
propranolol hydrochloride, fosinopril sodium, lidocaine
hydrochloride, eptifibatide, cefazolin sodium, atropine sulfate,
aminocaproic acid, spironolactone, interferon, sotalol
hydrochloride, potassium chloride, docusate sodium, dobutamine HCl,
alprazolam, pravastatin sodium, atorvastatin calcium, midazolam
hydrochloride, meperidine hydrochloride, isosorbide dinitrate,
epinephrine, dopamine hydrochloride, bivalirudin, rosuvastatin,
ezetimibe/simvastatin, avasimibe, and cariporide.
[0200] Non-limiting examples of therapeutic agents for psoriasis
with which a compound of Formula (I) can be combined include the
following: calcipotriene, clobetasol propionate, triamcinolone
acetonide, halobetasol propionate, tazarotene, methotrexate,
fluocinonide, betamethasone diprop augmented, fluocinolone
acetonide, acitretin, tar shampoo, betamethasone valerate,
mometasone furoate, ketoconazole, pramoxine/fluocinolone,
hydrocortisone valerate, flurandrenolide, urea, betamethasone,
clobetasol propionate/emoll, fluticasone propionate, azithromycin,
hydrocortisone, moisturizing formula, folic acid, desonide,
pimecrolimus, coal tar, diflorasone diacetate, etanercept folate,
lactic acid, methoxsalen, hc/bismuth subgal/znox/resor,
methylprednisolone acetate, prednisone, sunscreen, halcinonide,
salicylic acid, anthralin, clocortolone pivalate, coal extract,
coal tar/salicylic acid, coal tar/salicylic acid/sulfur,
desoximetasone, diazepam, emollient, fluocinonide/emollient,
mineral oil/castor oil/na lact, mineral oil/peanut oil,
petroleum/isopropyl myristate, psoralen, salicylic acid,
soap/tribromsalan, thimerosal/boric acid, celecoxib, infliximab,
cyclosporine, alefacept, efalizumab, tacrolimus, pimecrolimus,
PUVA, UVB, sulfasalazine, ABT-874 and ustekinamab.
[0201] Non-limiting examples of therapeutic agents for psoriatic
arthritis with which a compound of Formula (I) can be combined
include the following: methotrexate, etanercept, rofecoxib,
celecoxib, folic acid, sulfasalazine, naproxen, leflunomide,
methylprednisolone acetate, indomethacin, hydroxychloroquine
sulfate, prednisone, sulindac, betamethasone diprop augmented,
infliximab, methotrexate, folate, triamcinolone acetonide,
diclofenac, dimethylsulfoxide, piroxicam, diclofenac sodium,
ketoprofen, meloxicam, methylprednisolone, nabumetone, tolmetin
sodium, calcipotriene, cyclosporine, diclofenac sodium/misoprostol,
fluocinonide, glucosamine sulfate, gold sodium thiomalate,
hydrocodone bitartrate/apap, ibuprofen, risedronate sodium,
sulfadiazine, thioguanine, valdecoxib, alefacept, D2E7 (U.S. Pat.
No. 6,090,382, HUMIRA.TM.), and efalizumab.
[0202] Non-limiting examples of therapeutic agents for restenosis
with which a compound of Formula (I) can be combined include the
following: sirolimus, paclitaxel, everolimus, tacrolimus, ABT-578,
and acetaminophen.
[0203] Non-limiting examples of therapeutic agents for sciatica
with which a compound of Formula (I) can be combined include the
following: hydrocodone bitartrate/apap, rofecoxib, cyclobenzaprine
HCl, methylprednisolone, naproxen, ibuprofen, oxycodone
HCl/acetaminophen, celecoxib, valdecoxib, methylprednisolone
acetate, prednisone, codeine phosphate/apap, tramadol
HCl/acetaminophen, metaxalone, meloxicam, methocarbamol, lidocaine
hydrochloride, diclofenac sodium, gabapentin, dexamethasone,
carisoprodol, ketorolac tromethamine, indomethacin, acetaminophen,
diazepam, nabumetone, oxycodone HCl, tizanidine HCl, diclofenac
sodium/misoprostol, propoxyphene n-pap, asa/oxycod/oxycodone ter,
ibuprofen/hydrocodone bit, tramadol HCl, etodolac, propoxyphene
HCl, amitriptyline HCl, carisoprodol/codeine phos/asa, morphine
sulfate, multivitamins, naproxen sodium, orphenadrine citrate, and
temazepam.
[0204] Preferred examples of therapeutic agents for SLE (Lupus)
with which a compound of Formula (I) can be combined include the
following: NSAIDS, for example, diclofenac, naproxen, ibuprofen,
piroxicam, indomethacin; COX2 inhibitors, for example, celecoxib,
rofecoxib, valdecoxib; anti-malarials, for example,
hydroxychloroquine; steroids, for example, prednisone,
prednisolone, budenoside, dexamethasone; cytotoxics, for example,
azathioprine, cyclophosphamide, mycophenolate mofetil,
methotrexate; inhibitors of PDE4 or purine synthesis inhibitor, for
example Cellcept.RTM.. A compound of Formula (I) may also be
combined with agents such as sulfasalazine, 5-aminosalicylic acid,
olsalazine, Imuran.RTM. and agents which interfere with synthesis,
production or action of proinflammatory cytokines such as IL-1, for
example, caspase inhibitors like IL-1.beta. converting enzyme
inhibitors and IL-1ra. A compound of Formula (I) may also be used
with T cell signaling inhibitors, for example, tyrosine kinase
inhibitors; or molecules that target T cell activation molecules,
for example, CTLA-4-IgG or anti-B7 family antibodies, anti-PD-1
family antibodies. A compound of Formula (I) (can be combined with
IL-11 or anti-cytokine antibodies, for example, fonotolizumab
(anti-IFNg antibody), or anti-receptor receptor antibodies, for
example, anti-IL-6 receptor antibody and antibodies to B-cell
surface molecules. A compound of Formula (I) may also be used with
LJP 394 (abetimus), agents that deplete or inactivate B-cells, for
example, Rituximab (anti-CD20 antibody), lymphostat-B (anti-BlyS
antibody), TNF antagonists, for example, anti-TNF antibodies, D2E7
(U.S. Pat. No. 6,090,382; HUMIRA.TM.), CA2 (REMICADE.TM.), CDP 571,
TNFR-Ig constructs, (p75TNFRIgG (ENBREL.TM.) and p55TNFRIgG
(LENERCEPT.TM.).
[0205] In this invention, the following definitions are
applicable:
[0206] A "therapeutically effective amount" is an amount of a
compound of Formula (I) or a combination of two or more such
compounds, which inhibits, totally or partially, the progression of
the condition or alleviates, at least partially, one or more
symptoms of the condition. A therapeutically effective amount can
also be an amount which is prophylactically effective. The amount
which is therapeutically effective will depend upon the patient's
size and gender, the condition to be treated, the severity of the
condition and the result sought. For a given patient, a
therapeutically effective amount can be determined by methods known
to those of skill in the art.
[0207] "Pharmaceutically acceptable salts" refers to those salts
which retain the biological effectiveness and properties of the
free bases and which are obtained by reaction with inorganic acids,
for example, hydrochloric acid, hydrobromic acid, sulfuric acid,
nitric acid, and phosphoric acid or organic acids such as sulfonic
acid, carboxylic acid, organic phosphoric acid, methanesulfonic
acid, ethanesulfonic acid, p-toluenesulfonic acid, citric acid,
fumaric acid, maleic acid, succinic acid, benzoic acid, salicylic
acid, lactic acid, tartaric acid (e.g. (+) or (-)-tartaric acid or
mixtures thereof), amino acids (e.g. (+) or (-)-amino acids or
mixtures thereof), and the like. These salts can be prepared by
methods known to those skilled in the art.
[0208] Certain compounds of Formula (I) which have acidic
substituents may exist as salts with pharmaceutically acceptable
bases. The present invention includes such salts. Examples of such
salts include sodium salts, potassium salts, lysine salts and
arginine salts. These salts may be prepared by methods known to
those skilled in the art.
[0209] Certain compounds of Formula (I) and their salts may exist
in more than one crystal form and the present invention includes
each crystal form and mixtures thereof.
[0210] Certain compounds of Formula (I) and their salts may also
exist in the form of solvates, for example hydrates, and the
present invention includes each solvate and mixtures thereof.
[0211] Certain compounds of Formula (I) may contain one or more
chiral centers, and exist in different optically active forms. When
compounds of Formula (I) contain one chiral center, the compounds
exist in two enantiomeric forms and the present invention includes
both enantiomers and mixtures of enantiomers, such as racemic
mixtures. The enantiomers may be resolved by methods known to those
skilled in the art, for example by formation of diastereoisomeric
salts which may be separated, for example, by crystallization;
formation of diastereoisomeric derivatives or complexes which may
be separated, for example, by crystallization, gas-liquid or liquid
chromatography; selective reaction of one enantiomer with an
enantiomer-specific reagent, for example enzymatic esterification;
or gas-liquid or liquid chromatography in a chiral environment, for
example on a chiral support for example silica with a bound chiral
ligand or in the presence of a chiral solvent. It will be
appreciated that where the desired enantiomer is converted into
another chemical entity by one of the separation procedures
described above, a further step is required to liberate the desired
enantiomeric form. Alternatively, specific enantiomers may be
synthesized by asymmetric synthesis using optically active
reagents, substrates, catalysts or solvents, or by converting one
enantiomer into the other by asymmetric transformation.
[0212] When a compound of Formula (I) contains more than one chiral
center, it may exist in diastereoisomeric forms. The
diastereoisomeric compounds may be separated by methods known to
those skilled in the art, for example chromatography or
crystallization and the individual enantiomers may be separated as
described above. The present invention includes each
diastereoisomer of compounds of Formula (I) (and mixtures
thereof.
[0213] Certain compounds of Formula (I) may exist in different
tautomeric forms or as different geometric isomers, and the present
invention includes each tautomer and/or geometric isomer of
compounds of Formula (I) and mixtures thereof.
[0214] Certain compounds of Formula (I) may exist in different
stable conformational forms which may be separable. Torsional
asymmetry due to restricted rotation about an asymmetric single
bond, for example because of steric hindrance or ring strain, may
permit separation of different conformers. The present invention
includes each conformational isomer of compounds of Formula (I) and
mixtures thereof.
[0215] Certain compounds of Formula (I) may exist in zwitterionic
form and the present invention includes each zwitterionic form of
compounds of Formula (I) (and mixtures thereof.
[0216] As used herein the term "pro-drug" refers to an agent which
is converted into the parent drug in vivo by some physiological
chemical process (e.g., a prodrug on being brought to the
physiological pH is converted to the desired drug form). Pro-drugs
are often useful because, in some situations, they may be easier to
administer than the parent drug. They may, for instance, be
bioavailable by oral administration whereas the parent drug is not.
The pro-drug may also have improved solubility in pharmacological
compositions over the parent drug. An example, without limitation,
of a pro-drug would be a compound of the present invention wherein
it is administered as an ester (the "pro-drug") to facilitate
transmittal across a cell membrane where water solubility is not
beneficial, but then it is metabolically hydrolyzed to the
carboxylic acid once inside the cell where water solubility is
beneficial.
[0217] Pro-drugs have many useful properties. For example, a
pro-drug may be more water soluble than the ultimate drug, thereby
facilitating intravenous administration of the drug. A pro-drug may
also have a higher level of oral bioavailability than the ultimate
drug. After administration, the prodrug is enzymatically or
chemically cleaved to deliver the ultimate drug in the blood or
tissue.
[0218] Exemplary pro-drugs upon cleavage release the corresponding
free acid, and such hydrolyzable ester-forming residues of the
compounds of this invention include but are not limited to
carboxylic acid substituents wherein the free hydrogen is replaced
by (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.12)alkanoyloxymethyl,
(C.sub.4-C.sub.9) 1-(alkanoyloxy)ethyl,
1-methyl-1-(alkanoyloxy)-ethyl having from 5 to 10 carbon atoms,
alkoxycarbonyloxymethyl having from 3 to 6 carbon atoms,
1-(alkoxycarbonyloxy)ethyl having from 4 to 7 carbon atoms,
1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8 carbon
atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon
atoms, 1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10 carbon
atoms, 3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl,
di-N,N--(C.sub.1-C.sub.2)alkylamino(C.sub.2-C.sub.3)alkyl (such as
.beta.-dimethylaminoethyl), carbamoyl-(C.sub.1-C.sub.2)alkyl,
N,N-di(C.sub.1-C.sub.2)-alkylcarbamoyl-(C.sub.1-C.sub.2)alkyl and
piperidino-, pyrrolidino- or morpholino(C.sub.2-C.sub.3)alkyl.
[0219] Other exemplary pro-drugs release an alcohol of Formula (I)
wherein the free hydrogen of the hydroxyl substituent (e.g.,
R.sup.1 contains hydroxyl) is replaced by
(C.sub.1-C.sub.6)alkanoyloxymethyl,
1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
1-methyl-1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
(C.sub.1-C.sub.12)alkoxycarbonyloxymethyl,
N--(C.sub.1-C.sub.6)alkoxycarbonylamino-methyl, succinoyl,
(C.sub.1-C.sub.6)alkanoyl, .alpha.-amino(C.sub.1-C.sub.4)alkanoyl,
arylacyl and .alpha.-aminoacyl, or
.alpha.-aminoacyl-.alpha.-aminoacyl wherein said .alpha.-aminoacyl
moieties are independently any of the naturally occurring L-amino
acids found in proteins, P(O)(OH).sub.2,
--P(O)(O(C.sub.1-C.sub.6)alkyl).sub.2 or glycosyl (the radical
resulting from detachment of the hydroxyl of the hemiacetal of a
carbohydrate).
[0220] The term "heterocyclic," "heterocyclyl" or
"heterocyclylene," as used herein, include non-aromatic, ring
systems, including, but not limited to, monocyclic, bicyclic,
tricyclic and spirocyclic rings, which can be completely saturated
or which can contain one or more units of unsaturation, for the
avoidance of doubt, the degree of unsaturation does not result in
an aromatic ring system) and have 5 to 12 atoms including at least
one heteroatom, such as nitrogen, oxygen, or sulfur. For purposes
of exemplification, which should not be construed as limiting the
scope of this invention, the following are examples of heterocyclic
rings: azepinyl, azetidinyl, indolinyl, isoindolinyl, morpholinyl,
piperazinyl, piperidinyl, pyrrolidinyl, quinuclidinyl,
thiomorpholinyl, tetrahydropyranyl, tetrahydrofuranyl,
tetrahydroindolyl, thiomorpholinyl and tropanyl.
[0221] The term "heteroaryl" or "heteroarylene" as used herein,
include aromatic ring systems, including, but not limited to,
monocyclic, bicyclic and tricyclic rings, and have 5 to 12 atoms
including at least one heteroatom, such as nitrogen, oxygen, or
sulfur. For purposes of exemplification, which should not be
construed as limiting the scope of this invention: azaindolyl,
benzo(b)thienyl, benzimidazolyl, benzofuranyl, benzoxazolyl,
benzothiazolyl, benzothiadiazolyl, benzoxadiazolyl, furanyl,
imidazolyl, imidazopyridinyl, indolyl, indazolyl, isoxazolyl,
isothiazolyl, oxadiazolyl, oxazolyl, purinyl, pyranyl, pyrazinyl,
pyrazolyl, pyridinyl, pyrimidinyl, pyrrolyl,
pyrrolo[2,3-d]pyrimidinyl, pyrazolo[3,4-d]pyrimidinyl, quinolinyl,
quinazolinyl, triazolyl, thiazolyl, thiophenyl, tetrazolyl,
thiadiazolyl, thienyl, 6H-pyrrolo
[2,3-e][1,2,4]triazolo[4,3-a]pyrazinyl,
6H-imidazo[1,5-a]pyrrolo[2,3-e]pyrazinyl,
1,6-dihydropyrazolo[3,4-d]pyrrolo[2,3-b]pyridine,
3H-3,4,6,8a-tetraaza-asindacenyl, 3H-imidazo
[1,2-a]pyrrolo[2,3-e]pyrazinyl,
pyrazolo[3,4-d]pyrrolo[2,3-b]pyridinyl,
1,6-dihydro-1,2,5,6-tetraza-as-indacenyl,
3H-3,4,8a-triaza-as-indacenyl, 6H-3-oxa-2,5,6-triaza-as-indacenyl,
3,6-dihydro-2,3,6-tetraaza-as-indacenyl,
1,6-dihydro-dipyrrolo[2,3-b; 2'3'-d]pyridinyl,
6H-3-thia-2,5,6-triaza-as-indacenyl,
4,5-dihydro-1H-benzo[b]azepin-2(3H)-one,
3,4-dihydroquinolin-2(1H)-one, 2H-benzo[b][1,4]oxazin-3(4H)-one, or
6,7-dihydro-4H-pyrazolo [5,1-c][1,4]oxazinyl or
1,6-dihydroimidazo[4,5-d]pyrrolo[2,3-b]pyridine.
[0222] As used herein, "alkyl," "alkylene" or notations such as
"(C.sub.1-C.sub.8)" include straight chained or branched
hydrocarbons which are completely saturated. Examples of alkyls are
methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl and isomers
thereof. As used herein, "alkenyl," "alkenylene," "alkynylene" and
"alkynyl" means C.sub.2-C.sub.8 and includes straight chained or
branched hydrocarbons which contain one or more units of
unsaturation, one or more double bonds for alkenyl and one or more
triple bonds for alkynyl.
[0223] As used herein, "aromatic" groups (or "aryl" or "arylene"
groups) include aromatic carbocyclic ring systems (e.g. phenyl) and
fused polycyclic aromatic ring systems (e.g. naphthyl, biphenyl and
1,2,3,4-tetrahydronaphthyl).
[0224] As used herein, "cycloalkyl" or "cycloalkylene" means
C.sub.3-C.sub.12 monocyclic or multicyclic (e.g., bicyclic,
tricyclic, spirocyclic, etc.) hydrocarbons that is completely
saturated. Examples of a cycloalkyl group are cyclopropyl,
cyclobutyl, cyclopentyl, bicyclo[1.1.1]pentyl, and cyclohexyl.
[0225] As used herein, "cycloalkenyl" means C.sub.3-C.sub.12
monocyclic or multicyclic (e.g., bicyclic, tricyclic, spirocyclic,
etc.) hydrocarbons that has one or more unsaturated bonds but does
not amount to an aromatic group. Examples of a cycloalkenyl group
are cyclopentenyl and cyclohexenyl.
[0226] As used herein, many moieties or substituents are termed as
being either "substituted" or "optionally substituted". When a
moiety is modified by one of these terms, unless otherwise noted,
it denotes that any portion of the moiety that is known to one
skilled in the art as being available for substitution can be
substituted, which includes one or more substituents, where if more
than one substituent then each substituent is independently
selected. Such means for substitution are well-known in the art
and/or taught by the instant disclosure. For purposes of
exemplification, which should not be construed as limiting the
scope of this invention, some examples of groups that are
substituents are: (C.sub.1-C.sub.8)alkyl groups,
(C.sub.2-C.sub.8)alkenyl groups, (C.sub.2-C.sub.8)alkynyl groups,
(C.sub.3-C.sub.10)cycloalkyl groups, halogen (F, Cl, Br or I),
halogenated (C.sub.1-C.sub.8)alkyl groups (for example but not
limited to --CF.sub.3), --O--(C.sub.1-C.sub.8)alkyl groups, .dbd.O,
.dbd.CH.sub.2, --OH, --CH.sub.2OH, --CH.sub.2NH.sub.2,
(C.sub.1-C.sub.4)alkyl-OH, --CH.sub.2CH.sub.2OCH.sub.2CH.sub.3,
--S--(C.sub.1-C.sub.8)alkyl groups, --SH,
--NH(C.sub.1-C.sub.8)alkyl groups,
--N((C.sub.1-C.sub.8)alkyl).sub.2 groups, --NH.sub.2,
--C(O)NH.sub.2, --CH.sub.2NHC(O)(C.sub.1-C.sub.4)alkyl,
--CH.sub.2NHC(O)CH.sub.2Cl, --CH.sub.2NHC(O)CH.sub.2CN,
--CH.sub.2NHC(O)CH.sub.2CH.sub.2N(CH.sub.3).sub.2,
--CH.sub.2NHC(O)C(.dbd.CH.sub.2)CH.sub.3,
--CH.sub.2NHC(O)(C.sub.2-C.sub.4)alkynyl,
--CH.sub.2NHC(O)CH.sub.2CH.sub.2-piperidinyl,
--(C.sub.1-C.sub.4)alkyl-morpholinyl,
--CH.sub.2NHC(O)CH.sub.2O-phenyl wherein the phenyl is optionally
substituted with halogen, (C.sub.1-C.sub.4)alkoxy,
--C(O)(C.sub.1-C.sub.4)alkyl, --C(O)(C.sub.1-C.sub.4)alkoxy,
--C(O)N(H).sub.2, --C(O)N(CH.sub.3).sub.2,
--C(O)(C.sub.1-C.sub.6)heteroaryl, --N(CH.sub.3).sub.2,
--NHC(O)(C.sub.1-C.sub.4)alkyl, --NHC(O)(C.sub.2-C.sub.4)alkenyl,
--NHC(O)CH.sub.2CN, --S(O).sub.2(C.sub.1-C.sub.4)alkyl,
--S(O).sub.2(C.sub.1-C.sub.6)heteroaryl,
--S(O).sub.2(C.sub.1-C.sub.6) (C.sub.1-C.sub.6)heterocyclyl,
4-methylpiperazinecarbonyl, --(C.sub.1-C.sub.4)alkylC(O)NH.sub.2,
--C(O)NH(C.sub.1-C.sub.8)alkyl groups,
--C(O)N((C.sub.1-C.sub.8)alkyl).sub.2,
--C(O)N(H)(C.sub.3-C.sub.8)cycloalkyl groups,
--C(O)(C.sub.1-C.sub.4)alkoxy, --NHC(O)H,
--NHC(O)(C.sub.1-C.sub.8)alkyl groups,
--NHC(O)(C.sub.3-C.sub.8)cycloalkyl groups,
--N((C.sub.1-C.sub.8)alkyl)C(O)H,
--N((C.sub.1-C.sub.8)alkyl)C(O)(C.sub.1-C.sub.8)alkyl groups,
--NHC(O)NH.sub.2, --NHC(O)NH(C.sub.1-C.sub.8)alkyl groups,
--N((C.sub.1-C.sub.8)alkyl)C(O)NH.sub.2 groups,
--NHC(O)N((C.sub.1-C.sub.8)alkyl).sub.2 groups,
--N((C.sub.1-C.sub.8)alkyl)C(O)N((C.sub.1-C.sub.8)alkyl).sub.2
groups, --N((C.sub.1-C.sub.8)alkyl)C(O)NH((C.sub.1-C.sub.8)alkyl),
--NHCH.sub.2-- heteroaryl, benzyl, --OCH.sub.2-heteroaryl, --C(O)H,
--C(O)(C.sub.1-C.sub.8)alkyl groups, --CN, --NO.sub.2,
S(O)(C.sub.1-C.sub.8)alkyl groups,
--S(O).sub.2(C.sub.1-C.sub.8)alkyl groups,
--S(O).sub.2N((C.sub.1-C.sub.8)alkyl).sub.2 groups,
S(O).sub.2NH(C.sub.1-C.sub.6)alkyl groups,
--S(O).sub.2NH(C.sub.3-C.sub.8)cycloalkyl groups,
--S(O).sub.2NH.sub.2 groups, --NHS(O).sub.2(C.sub.1-C.sub.6)alkyl
groups, --N((C.sub.1-C.sub.8)alkyl)S(O).sub.2(C.sub.1-C.sub.6)alkyl
groups, --(C.sub.1-C.sub.8)alkyl-O--(C.sub.1-C.sub.8)alkyl groups,
--O--(C.sub.1-C.sub.8)alkyl-O--(C.sub.1-C.sub.8)alkyl groups,
--C(O)OH, --C(O)O(C.sub.1-C.sub.8)alkyl groups, NHOH,
NHO(C.sub.1-C.sub.8)alkyl groups, --O-halogenated
(C.sub.1-C.sub.8)alkyl groups (for example but not limited to
--OCF.sub.3), --S(O).sub.2-halogenated (C.sub.1-C.sub.8)alkyl
groups (for example but not limited to --S(O).sub.2CF.sub.3),
--S-halogenated (C.sub.1-C.sub.8)alkyl groups (for example but not
limited to --SCF.sub.3), --(C.sub.1-C.sub.6)heterocyclyl (for
example but not limited to pyrrolidine, tetrahydrofuran, pyran or
morpholine), --(C.sub.1-C.sub.6)heteroaryl (for example but not
limited to tetrazole, imidazole, furan, pyrazine or pyrazole),
-phenyl, optionally substituted benzyl,
--NHC(O)O--(C.sub.1-C.sub.6)alkyl groups,
--N((C.sub.1-C.sub.6)alkyl)C(O)O--(C.sub.1-C.sub.6)alkyl groups,
--C(.dbd.NH)--(C.sub.1-C.sub.6)alkyl groups,
--C(.dbd.NOH)--(C.sub.1-C.sub.6)alkyl groups, or
--C(.dbd.N--O--(C.sub.1-C.sub.6)alkyl)-(C.sub.1-C.sub.6)alkyl
groups.
[0227] The term "kit" as used herein refers to a packaged product
comprising components with which to administer a compound of
Formula (I) of the invention for treatment of an autoimmune
disorder. The kit preferably comprises a box or container that
holds the components of the kit. The box or container is affixed
with a label or a Food and Drug Administration approved protocol.
The box or container holds components of the invention which are
preferably contained within plastic, polyethylene, polypropylene,
ethylene, or propylene vessels. The vessels can be capped-tubes or
bottles. The kit can also include instructions for administering a
compound of Formula (I).
[0228] One or more compounds of this invention can be administered
to a human patient by themselves or in pharmaceutical compositions
where they are mixed with biologically suitable carriers or
excipient(s) at doses to treat or ameliorate a disease or condition
as described herein. Mixtures of these compounds can also be
administered to the patient as a simple mixture or in suitable
formulated pharmaceutical compositions. A therapeutically effective
dose refers to that amount of the compound or compounds sufficient
to result in the prevention or attenuation of a disease or
condition as described herein. Techniques for formulation and
administration of the compounds of the instant application may be
found in references well known to one of ordinary skill in the art,
such as "Remington's Pharmaceutical Sciences," Mack Publishing Co.,
Easton, Pa., latest edition.
[0229] Suitable routes of administration may, for example, include
oral, eyedrop, rectal, transmucosal, topical, or intestinal
administration; parenteral delivery, including intramuscular,
subcutaneous, intramedullary injections, as well as intrathecal,
direct intraventricular, intravenous, intrapcritoncal, intranasal,
or intraocular injections.
[0230] Alternatively, one may administer the compound in a local
rather than a systemic manner, for example, via injection of the
compound directly into an edematous site, often in a depot or
sustained release formulation.
[0231] Furthermore, one may administer the drug in a targeted drug
delivery system, for example, in a liposome coated with endothelial
cell-specific antibody.
[0232] The pharmaceutical compositions of the present invention may
be manufactured in a manner that is itself known, e.g., by means of
conventional mixing, dissolving, granulating, dragee-making,
levigating, emulsifying, encapsulating, entrapping or lyophilizing
processes.
[0233] Pharmaceutical compositions for use in accordance with the
present invention thus may be formulated in a conventional manner
using one or more physiologically acceptable carriers comprising
excipients and auxiliaries which facilitate processing of the
active compounds into preparations which can be used
pharmaceutically. Proper formulation is dependent upon the route of
administration chosen.
[0234] For injection, the agents of the invention may be formulated
in aqueous solutions, preferably in physiologically compatible
buffers such as Hanks' solution, Ringer's solution, or
physiological saline buffer. For transmucosal administration,
penetrants appropriate to the barrier to be permeated are used in
the formulation. Such penetrants are generally known in the
art.
[0235] For oral administration, the compounds can be formulated
readily by combining the active compounds with pharmaceutically
acceptable carriers well known in the art. Such carriers enable the
compounds of the invention to be formulated as tablets, pills,
dragees, capsules, liquids, gels, syrups, slurries, suspensions and
the like, for oral ingestion by a patient to be treated.
Pharmaceutical preparations for oral use can be obtained by
combining the active compound with a solid excipient, optionally
grinding a resulting mixture, and processing the mixture of
granules, after adding suitable auxiliaries, if desired, to obtain
tablets or dragee cores. Suitable excipients are, in particular,
fillers such as sugars, including lactose, sucrose, mannitol, or
sorbitol; cellulose preparations such as, for example, maize
starch, wheat starch, rice starch, potato starch, gelatin, gum
tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium
carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If
desired, disintegrating agents may be added, such as the
cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt
thereof such as sodium alginate.
[0236] Dragee cores are provided with suitable coatings. For this
purpose, concentrated sugar solutions may be used, which may
optionally contain gum arabic, talc, polyvinyl pyrrolidone,
carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer
solutions, and suitable organic solvents or solvent mixtures.
Dyestuffs or pigments may be added to the tablets or dragee
coatings for identification or to characterize different
combinations of active compound doses.
[0237] Pharmaceutical preparations that can be used orally include
push-fit capsules made of gelatin, as well as soft, sealed capsules
made of gelatin and a plasticizer, such as glycerol or sorbitol.
The push-fit capsules can contain the active ingredients in
admixture with filler such as lactose, binders such as starches,
and/or lubricants such as talc or magnesium stearate and,
optionally, stabilizers. In soft capsules, the active compounds may
be dissolved or suspended in suitable liquids, such as fatty oils,
liquid paraffin, or liquid polyethylene glycols. In addition,
stabilizers may be added. All formulations for oral administration
should be in dosages suitable for such administration.
[0238] For buccal administration, the compositions may take the
form of tablets or lozenges formulated in conventional manner.
[0239] For administration by inhalation, the compounds for use
according to the present invention are conveniently delivered in
the form of an aerosol spray presentation from pressurized packs or
a nebuliser, with the use of a suitable propellant, e.g.,
dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In
the case of pressurized aerosol the dosage unit may be determined
by providing a valve to deliver a metered amount. Capsules and
cartridges of e.g. gelatin for use in an inhaler or insufflator may
be formulated containing a powder mix of the compound and a
suitable powder base such as lactose or starch.
[0240] The compounds can be formulated for parenteral
administration by injection, e.g. bolus injection or continuous
infusion. Formulations for injection may be presented in unit
dosage form, e.g. in ampoules or in multi-dose containers, with an
added preservative. The compositions may take such forms as
suspensions, solutions or emulsions in oily or aqueous vehicles,
and may contain formulatory agents such as suspending, stabilizing
and/or dispersing agents.
[0241] Pharmaceutical formulations for parenteral administration
include aqueous solutions of the active compounds in water-soluble
form. Additionally, suspensions of the active compounds may be
prepared as appropriate oily injection suspensions. Suitable
lipophilic solvents or vehicles include fatty oils such as sesame
oil, or synthetic fatty acid esters, such as ethyl oleate or
triglycerides, or liposomes. Aqueous injection suspensions may
contain substances which increase the viscosity of the suspension,
such as sodium carboxymethyl cellulose, sorbitol, or dextran.
Optionally, the suspension may also contain suitable stabilizers or
agents which increase the solubility of the compounds to allow for
the preparation of highly concentrated solutions.
[0242] Alternatively, the active ingredient may be in powder form
for constitution with a suitable vehicle, e.g., sterile
pyrogen-free water, before use.
[0243] The compounds may also be formulated in rectal compositions
such as suppositories or retention enemas, e.g., containing
conventional suppository bases such as cocoa butter or other
glycerides.
[0244] In addition to the formulations described previously, the
compounds may also be formulated as a depot preparation. Such long
acting formulations may be administered by implantation (for
example subcutaneously or intramuscularly or by intramuscular
injection). Thus, for example, the compounds may be formulated with
suitable polymeric or hydrophobic materials (for example as an
emulsion in an acceptable oil) or ion exchange resins, or as
sparingly soluble derivatives, for example, as a sparingly soluble
salt.
[0245] An example of a pharmaceutical carrier for the hydrophobic
compounds of the invention is a cosolvent system comprising benzyl
alcohol, a nonpolar surfactant, a water-miscible organic polymer,
and an aqueous phase. The cosolvent system may be the VPD
co-solvent system. VPD is a solution of 3% w/v benzyl alcohol, 8%
w/v of the nonpolar surfactant polysorbate 80, and 65% w/v
polyethylene glycol 300, made up to volume in absolute ethanol. The
VPD co-solvent system (VPD:5W) consists of VPD diluted 1:1 with a
5% dextrose in water solution. This co-solvent system dissolves
hydrophobic compounds well, and itself produces low toxicity upon
systemic administration. Naturally, the proportions of a co-solvent
system may be varied considerably without destroying its solubility
and toxicity characteristics. Furthermore, the identity of the
co-solvent components may be varied: for example, other
low-toxicity nonpolar surfactants may be used instead of
polysorbate 80; the fraction size of polyethylene glycol may be
varied; other biocompatible polymers may replace polyethylene
glycol, e.g. polyvinyl pyrrolidone; and other sugars or
polysaccharides may substitute for dextrose.
[0246] Alternatively, other delivery systems for hydrophobic
pharmaceutical compounds may be employed. Liposomes and emulsions
are well known examples of delivery vehicles or carriers for
hydrophobic drugs. Certain organic solvents such as
dimethylsulfoxide also may be employed, although usually at the
cost of greater toxicity. Additionally, the compounds may be
delivered using a sustained-release system, such as semipermeable
matrices of solid hydrophobic polymers containing the therapeutic
agent. Various sustained-release materials have been established
and are well known by those skilled in the art. Sustained-release
capsules may, depending on their chemical nature, release the
compounds for a few weeks up to over 100 days. Depending on the
chemical nature and the biological stability of the therapeutic
reagent, additional strategies for protein stabilization may be
employed.
[0247] The pharmaceutical compositions also may comprise suitable
solid or gel phase carriers or excipients. Examples of such
carriers or excipients include but are not limited to calcium
carbonate, calcium phosphate, various sugars, starches, cellulose
derivatives, gelatin, and polymers such as polyethylene
glycols.
[0248] Many of the compounds of the invention may be provided as
salts with pharmaceutically compatible counter ions.
Pharmaceutically compatible salts may be formed with many acids,
including but not limited to hydrochloric, sulfuric, acetic,
lactic, tartaric, malic, succinic, etc. Salts tend to be more
soluble in aqueous or other protonic solvents than are the
corresponding free base forms.
[0249] Pharmaceutical compositions suitable for use in the present
invention include compositions wherein the active ingredients are
contained in an effective amount to achieve its intended purpose.
More specifically, a therapeutically effective amount means an
amount effective to prevent development of or to alleviate the
existing symptoms of the subject being treated. Determination of
the effective amounts is well within the capability of those
skilled in the art.
[0250] For any compound used in a method of the present invention,
the therapeutically effective dose can be estimated initially from
cellular assays. For example, a dose can be formulated in cellular
and animal models to achieve a circulating concentration range that
includes the IC.sub.50 as determined in cellular assays (e.g., the
concentration of the test compound which achieves a half-maximal
inhibition of a given protein kinase activity). In some cases it is
appropriate to determine the IC.sub.50 in the presence of 3 to 5%
serum albumin since such a determination approximates the binding
effects of plasma protein on the compound. Such information can be
used to more accurately determine useful doses in humans. Further,
the most preferred compounds for systemic administration
effectively inhibit protein kinase signaling in intact cells at
levels that are safely achievable in plasma.
[0251] A therapeutically effective dose refers to that amount of
the compound that results in amelioration of symptoms in a patient.
Toxicity and therapeutic efficacy of such compounds can be
determined by standard pharmaceutical procedures in cell cultures
or experimental animals, e.g., for determining the maximum
tolerated dose (MTD) and the ED.sub.50 (effective dose for 50%
maximal response). The dose ratio between toxic and therapeutic
effects is the therapeutic index and it can be expressed as the
ratio between MTD and ED.sub.50. Compounds which exhibit high
therapeutic indices are preferred. The data obtained from these
cell culture assays and animal studies can be used in formulating a
range of dosage for use in humans. The dosage of such compounds
lies preferably within a range of circulating concentrations that
include the ED.sub.50 with little or no toxicity. The dosage may
vary within this range depending upon the dosage form employed and
the route of administration utilized. The exact formulation, route
of administration and dosage can be chosen by the individual
physician in view of the patient's condition (see, e.g., Fingl et
al., 1975, in The Pharmacological Basis of Therapeutics, Ch. 1, p.
1). In the treatment of crises, the administration of an acute
bolus or an infusion approaching the MTD may be required to obtain
a rapid response.
[0252] Dosage amount and interval may be adjusted individually to
provide plasma levels of the active moiety which are sufficient to
maintain the kinase modulating effects, or minimal effective
concentration (MEC). The MEC will vary for each compound but can be
estimated from in vitro data; e.g. the concentration necessary to
achieve 50-90% inhibition of protein kinase using the assays
described herein. Dosages necessary to achieve the MEC will depend
on individual characteristics and route of administration. However,
HPLC assays or bioassays can be used to determine plasma
concentrations.
[0253] Dosage intervals can also be determined using the MEC value.
Compounds should be administered using a regimen which maintains
plasma levels above the MEC for 10-90% of the time, preferably
between 30-90% and most preferably between 50-90% until the desired
amelioration of symptoms is achieved. In cases of local
administration or selective uptake, the effective local
concentration of the drug may not be related to plasma
concentration.
[0254] The amount of composition administered will, of course, be
dependent on the subject being treated, on the subject's weight,
the severity of the affliction, the manner of administration and
the judgment of the prescribing physician.
[0255] The compositions may, if desired, be presented in a pack or
dispenser device which may contain one or more unit dosage forms
containing the active ingredient. The pack may for example comprise
metal or plastic foil, such as a blister pack. The pack or
dispenser device may be accompanied by instructions for
administration. Compositions comprising a compound of the invention
formulated in a compatible pharmaceutical carrier may also be
prepared, placed in an appropriate container, and labelled for
treatment of an indicated condition.
[0256] In some formulations it may be beneficial to use the
compounds of the present invention in the form of particles of very
small size, for example as obtained by fluid energy milling.
[0257] The use of compounds of the present invention in the
manufacture of pharmaceutical compositions is illustrated by the
following description. In this description the term "active
compound" denotes any compound of the invention but particularly
any compound which is the final product of one of the following
Examples.
a) Capsules
[0258] In the preparation of capsules, 10 parts by weight of active
compound and 240 parts by weight of lactose can be de-aggregated
and blended. The mixture can be filled into hard gelatin capsules,
each capsule containing a unit dose or part of a unit dose of
active compound.
b) Tablets
[0259] Tablets can be prepared, for example, from the following
ingredients.
TABLE-US-00001 Parts by weight Active compound 10 Lactose 190 Maize
starch 22 Polyvinylpyrrolidone 10 Magnesium stearate 3
[0260] The active compound, the lactose and some of the starch can
be de-aggregated, blended and the resulting mixture can be
granulated with a solution of the polyvinylpyrrolidone in ethanol.
The dry granulate can be blended with the magnesium stearate and
the rest of the starch. The mixture is then compressed in a
tabletting machine to give tablets each containing a unit dose or a
part of a unit dose of active compound.
c) Enteric Coated Tablets
[0261] Tablets can be prepared by the method described in (b)
above. The tablets can be enteric coated in a conventional manner
using a solution of 20% cellulose acetate phthalate and 3% diethyl
phthalate in ethanol:dichloromethane (1:1).
d) Suppositories
[0262] In the preparation of suppositories, for example, 100 parts
by weight of active compound can be incorporated in 1300 parts by
weight of triglyceride suppository base and the mixture formed into
suppositories each containing a therapeutically effective amount of
active ingredient.
[0263] In the compositions of the present invention the active
compound may, if desired, be associated with other compatible
pharmacologically active ingredients. For example, the compounds of
this invention can be administered in combination with another
therapeutic agent that is known to treat a disease or condition
described herein. For example, with one or more additional
pharmaceutical agents that inhibit or prevent the production of
VEGF or angiopoietins, attenuate intracellular responses to VEGF or
angiopoietins, block intracellular signal transduction, inhibit
vascular hyperpermeability, reduce inflammation, or inhibit or
prevent the formation of edema or neovascularization. The compounds
of the invention can be administered prior to, subsequent to or
simultaneously with the additional pharmaceutical agent, whichever
course of administration is appropriate. The additional
pharmaceutical agents include, but are not limited to, anti-edemic
steroids, NSAIDS, ras inhibitors, anti-TNF agents, anti-IL1 agents,
antihistamines, PAF-antagonists, COX-1 inhibitors, COX-2
inhibitors, NO synthase inhibitors, Akt/PTB inhibitors, IGF-1R
inhibitors, PKC inhibitors, PI3 kinase inhibitors, calcineurin
inhibitors and immunosuppressants. The compounds of the invention
and the additional pharmaceutical agents act either additively or
synergistically. Thus, the administration of such a combination of
substances that inhibit angiogenesis, vascular hyperpermeability
and/or inhibit the formation of edema can provide greater relief
from the deleterious effects of a hyperproliferative disorder,
angiogenesis, vascular hyperpermeability or edema than the
administration of either substance alone. In the treatment of
malignant disorders combinations with antiproliferative or
cytotoxic chemotherapies or radiation are included in the scope of
the present invention.
[0264] The present invention also comprises the use of a compound
of Formula (I) as a medicament.
[0265] A further aspect of the present invention provides the use
of a compound of Formula (I) or a salt thereof in the manufacture
of a medicament for treating vascular hyperpermeability,
angiogenesis-dependent disorders, proliferative diseases and/or
disorders of the immune system in mammals, particularly human
beings.
[0266] The present invention also provides a method of treating
vascular hyperpermeability, inappropriate neovascularization,
proliferative diseases and/or disorders of the immune system which
comprises the administration of a therapeutically effective amount
of a compound of Formula (I) to a mammal, particularly a human
being, in need thereof.
TABLE-US-00002 ABBREVIATIONS AcOH Glacial acctic acid Boc.sub.2O
di-tert-Butyl-dicarbonate br broad BuOH Butanol n-BuOH 1-Butanol
t-BuOH 2-Methyl-2-propanol tert-butyl
2-Di-tert-butylphosphino-2',4',6'- XPhos trissopropylbiphenyl d
Doublet dd Doublet of doublets dba Dibenzylideneacetone DCE
1,2-Dichlorethane DCM Dichloromethane (methylene chloride) DEA
Diethylamine DIAD Diisoporpyl azodicarboxylate DIEA
N,N-Diisopropylethylamine DMA Dimethylacetamide DMAP
4-(Demethylamino)pyridine DME 1,2-Dimethylformamide DMF
N,N-Dimethylformamide DMSO Dimethyl sulfoxide dppf 1,1'-
Bis(diphenylphosphino)fcrroccnc equiv Equivalent(s) Et Ethyl EtOAc
Ethyl acetate Et.sub.2O Dicthyl ether EtOH Ethanol g Gram(s) h
Hour(s) HPLC High-pressure liquid chromatography Hz Hertz i-PrOH
Isopropyl alcohol n-PrOH 1-Propanol KOAc Potassium acetate KOt-Bu
Potassium tert-butoxide LC Liquid chromatography LDA
lithiumdiisopropylamide m Multiplet M Molar Me Methyl MeCN
Acetonitrile MeOH Methyl alcohol 2-MeTHF 2-Methyltetrahydrofuran
min Minute(s) mL Milliliter(s) mmol Millimole MS Mass spectrometry
N Normal MTBE Methyl tert-butyl ether NaOt-Bu Sodium tert-butoxide
NH.sub.4OAc Ammonium acetate NMP 1-Methyl-2-pyrrolidinone NMR
Nuclear magnetic resonance or Optical rotation Pd(OAc).sub.2
Palladium(II) acetate PE Petroleum ether pH -log[H.sup.+] PPh.sub.3
Triphenylphosphine ppm Parts per million prep Preparatory psi
Pounds per square inch R.sub.t Rentention time rt Room temperature
q Quartet S Singlet SFC Supercritical fluid chromatography t
Triplet tq triplet of quartets t- tertiary TEA Triethylamine TFA
Trifluoroacetic acid THF Tetrahydrofuran TLC Thin layer
chromatography USP United States Pharmacopeia UV Ultraviolet wt %
Weight percent Xantphos 4,5-Bis(diphenylphosphino)-
9,9-dimethylxanthene XPhos 2-Dicyclohexylphosphino-
2',4',6'-triisopropylbiphenyl
In Vitro BTK Kinase Activity Measured by Time-Resolved Fluorescence
Resonance Energy Transfer (trFRET) The in-house BTK corresponds to
recombinant human catalytic domain (aa 393-659), which was
expressed in SF9 cells with an N-terminal his tag and purified by
immobilized metal affinity chromatography. BTK was mixed with
peptide substrate (biotin-TYR1, Sequence:
Biotin-(Ahx)-GAEEEIYAAFFA-COOH, 0.4 .mu.M final) at varying
inhibitor concentrations in reaction buffer: 50 mM MOPSO pH 6.5, 10
mM MgCl.sub.2, 2 mM MnCl.sub.2, 2.5 mM DTT, 0.01% BSA, 0.1 mM
Na.sub.3VO.sub.4 and 0.01 mM ATP. After about 60 min incubation at
rt, the reaction was quenched by addition of EDTA (final
concentration: 100 mM) and developed by addition of detection
reagents (final approximate concentrations: 30 mM HEPES pH 7.0,
0.06% BSA, 0.006% Tween-20, 0.24 M KF, 80 ng/mL PT66K (europium
labeled anti-phosphotyrosine antibody cat #61T66KLB Cisbio,
Bedford, Mass.) and 0.6 .mu.g/mL SAXL (Phycolink
streptavidin-allophycocyanin acceptor, cat #PJ25S, Prozyme, San
Leandro, Calif.). The developed reaction was incubated in the dark
for about 60 min at rt, then read via a time-resolved fluorescence
detector (Rubystar, BMG) using a 337 nm laser for excitation and
monitoring emission wavelength at 665 nm. Within the linear range
of the assay, the observed signal at 665 nm was directly related to
phosphorylated product and can be used to calculate the IC.sub.50
values. In Vitro CSF-1R Kinase Activity Measured by Time-Resolved
Fluorescence Resonance Energy Transfer (trFRET) The CSF-1R
construct corresponds to recombinant human catalytic domain (aa
538-910), which was purchased from Invitrogen (cat #PV4092). CSF-1R
was mixed with peptide substrate (biotin-TYR1, Sequence:
Biotin-(Ahx)-GAEEEIYAAFFA-COOH, 4 .mu.M final) at varying inhibitor
concentrations in reaction buffer: 50 mM MOPSO pH 6.5, 10 mM
MgCl.sub.2, 2 mM MnCl.sub.2, 2.5 mM DTT, 0.01% BSA, 0.1 mM
Na.sub.3VO.sub.4 and 0.1 mM ATP. After about 60 min incubation at
rt, the reaction was quenched by addition of EDTA (final
concentration: 100 mM) and developed by addition of detection
reagents (final approximate concentrations: 30 mM HEPES pH 7.0,
0.06% BSA, 0.006% Tween-20, 0.24 M KF, 80 ng/mL PT66K (europium
labeled anti-phosphotyrosine antibody cat #61T66KLB Cisbio,
Bedford, Mass.) and 0.6 .mu.g/mL SAXL (Phycolink
streptavidin-allophycocyanin acceptor, cat #PJ25S, Prozyme, San
Leandro, Calif.). The developed reaction was incubated in the dark
for about 60 min at rt, then read via a time-resolved fluorescence
detector (Rubystar, BMG) using a 337 nm laser for excitation and
monitoring emission wavelength at 665 nm. Within the linear range
of the assay, the observed signal at 665 nm was directly related to
phosphorylated product and can be used to calculate the IC.sub.50
values.
[0267] For the purpose of the Tables and Examples below, the Btk or
CSF-1R IC.sub.50 values of each compound is expressed as follows:
A=a compound with IC.sub.50 less than 0.1 .mu.M, B=a compound with
IC.sub.50 within the range of 0.1 .mu.M to 1 .mu.M, and C=a
compound with a Btk IC.sub.50 within the range of 1 .mu.M to 50
.mu.M. NT=not tested
[0268] The teachings of all references, including journal articles,
patents and published patent applications, are incorporated herein
by reference in their entirety.
[0269] The following examples are for illustrative purposes and are
not to be construed as limiting the scope of the present
invention.
General Synthetic Schemes
[0270] Compounds of the invention may be prepared using synthetic
transformations such as those illustrated in Schemes I-IV. Starting
materials are commercially available, may be prepared by the
procedures described herein, by literature procedures, or by
procedures that would be well known to one skilled in the art of
organic chemistry (see, for example, Larock, R. C. "Comprehensive
Organic Transformations: A Guide to Functional Group Preparations,
2.sup.nd edition", 1999, Wiley-VCH or Greene, T. W. and Wuts, P. G.
M. "Protective Groups in Organic Synthesis, 3.sup.rd Edition",
1999, Wiley-Interscience). Methods for preparing
[1,2,4]triazolo[1,5-a]pyrazin-8-amine compounds of the invention
are illustrated in Scheme I.
6,8-Dibromo-[1,2,4]triazolo[1,5-a]pyrazine 1 is commercially
available (e.g. Ark Pharm) and can be reacted with amines via
displacement chemistry using conditions known to one skilled in the
art such as those described in General Procedure A or via
palladium-mediated chemistry as described in General Procedure B to
give compounds 2. Compounds 3 may be obtained from Suzuki coupling
reaction (for example General Procedure C) of compounds 2 with
commercially available boronic acids or boronates or with boronates
prepared from halides as described in Larock, R. C. (referenced
above). Further functionalization of
[1,2,4]triazolo[1,5-a]pyrazin-8-amines 3 can be performed, if
desired, using reactions known to one skilled in the art (for
example Larock, R. C. referenced above). For example,
triazolopyrazines 3 containing a double bond may be reduced to
saturated systems using hydrogenation conditions such as those
described in General Procedure E. In addition, amides can be
prepared from triazolopyrazines 3 containing a primary or secondary
amine (for example General Procedures F). Also, deprotection of
triazolopyrazines 3 containing a protected primary or secondary
amine can be performed using conditions such as those described in
Greene, T. W. and Wuts, P. G. M. referenced above or in General
Procedures D. For example, for R' containing a protecting group
(for example a Boc group), the protecting group can be removed to
yield the unprotected amine (for example General Procedure D) and
the deprotected compounds 3 may then be reacted further as
described above.
##STR00003##
[0271] Methods for preparing [1,2,4]triazolo[1,5-a]pyridin-8-amine
compounds of the invention are illustrated in Scheme II. Using
conditions such as those described in Example #1, Step A,
commercially available 2-amino-3-bromo-5-chloropyridine 4 (e.g. Ark
Pharm) can be reacted with DMF-DMA to give intermediate 5 which may
then be cyclized to give
8-bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine 6.
8-Bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine 6 can be reacted
with amines via palladium-mediated conditions known to one skilled
in the art such as those described in General Procedure B to give
compounds 7. Compounds 8 may be obtained from Suzuki coupling
reaction (for example General Procedure C) of compounds 7 with
commercially available boronic acids or boronates or with boronates
prepared from halides as described in Larock, R. C. (referenced
above) or General Procedure H. Alternatively, compounds 8 can be
synthesized by using a different route illustrated in Scheme IIa.
Using conditions such as those described in Example #4, Step A,
commercially available 5-bromo-3-chloropyridin-2-amine 12 (e.g. Ark
Pharm) can be converted to
6-bromo-8-chloro-[1,2,4]triazolo[1,5-a]pyridine 13 in a similar
manner as Step A in Example #1. Synthesis of compounds 14 may be
achieved by Suzuki coupling reaction of compound 13 with with
commercially available boronic acids or boronates or with boronates
prepared from halides as described in Larock, R. C. (referenced
above) or General Procedure H. Compounds 14 can react with amines
via palladium-mediated conditions known to one skilled in the art
such as those described in General Procedure B to give compounds 8.
Further functionalization of [1,2,4]triazolo[1,5-a]pyridin-8-amine
8 can be performed, if desired, using reactions known to one
skilled in the art (for example Larock, R. C. referenced above).
For example, triazolopyridines 8 containing a double bond may be
reduced to saturated systems using hydrogenation conditions such as
those described in General Procedure E. In addition, amides can be
prepared from triazolopyridines 8 containing a primary or secondary
amine (for example General Procedures F). Also, deprotection of
triazolopyridines 8 containing a protected primary or secondary
amine can be performed using conditions such as those described in
Greene, T. W. and Wuts, P. G. M. referenced above or in General
Procedures D. For example, for R' containing a protecting group
(for example a Boc group), the protecting group can be removed to
yield the unprotected amine (for example General Procedure D) and
the deprotected compounds 8 may then be reacted further as
described above.
##STR00004##
##STR00005##
[0272] Methods for preparing [1,2,4]triazolo[1,5-a]pyrazin-8-amine
compounds of the invention are illustrated in Scheme III.
6,8-Dibromoimidazo[1,2-a]pyrazine 9 is commercially available (e.g.
Ark Pharm) and can be reacted with amines via displacement
chemistry using conditions known to one skilled in the art such as
those described in General Procedure A or via palladium-mediated
chemistry as described in General Procedure B to give compounds 10.
Compounds 11 may be obtained from Suzuki coupling reaction (for
example General Procedure C) of compounds 10 with commercially
available boronic acids or boronates or with boronates prepared
from halides as described in Larock, R. C. (referenced above).
Further functionalization of imidazo[1,2-a]pyrazin-8-amines 11 can
be performed, if desired, using reactions known to one skilled in
the art (for example Larock, R. C. referenced above). For example,
imidazopyrazines 11 containing a double bond may be reduced to
saturated systems using hydrogenation conditions such as those
described in General Procedure E. In addition, amides can be
prepared from imidazopyrazines 11 containing a primary or secondary
amine (for example General Procedures F). Also, deprotection of
imidazopyrazines 11 containing a protected primary or secondary
amine can be performed using conditions such as those described in
Greene, T. W. and Wuts, P. G. M. referenced above or in General
Procedures D. For example, for R' containing a protecting group
(for example a Boc group), the protecting group can be removed to
yield the unprotected amine (for example General Procedure D) and
the deprotected compounds 11 may then be reacted further as
described above.
##STR00006##
[0273] Methods for preparing imidazo[1,2-b]pyridazine compounds of
the invention are illustrated in Scheme III.
8-Bromo-6-chloroimidazo[1,2-b]pyridazine 12 is commercially
available (e.g. Astatech) and can be reacted with amines via
displacement chemistry using conditions known to one skilled in the
art such as those described in General Procedure A or via
palladium-mediated chemistry as described in General Procedure B to
give compounds 13. Compounds 14 may be obtained from Suzuki
coupling reaction (for example General Procedure C) of compounds 13
with commercially available boronic acids or boronates or with
boronates prepared from halides as described in Larock, R. C.
(referenced above). Further functionalization of
imidazo[1,2-b]pyridazin-8-amines 14 can be performed, if desired,
using reactions known to one skilled in the art (for example
Larock, R. C. referenced above). For example, imidazopyridazines 14
containing a double bond may be reduced to saturated systems using
hydrogenation conditions such as those described in General
Procedure E. In addition, amides can be prepared from
imidazopyridazines 14 containing a primary or secondary amine (for
example General Procedures F). Also, deprotection of
imidazopyridazines 14 containing a protected primary or secondary
amine can be performed using conditions such as those described in
Greene, T. W. and Wuts, P. G. M. referenced above or in General
Procedures D. For example, for R' containing a protecting group
(for example a Boc group), the protecting group can be removed to
yield the unprotected amine (for example General Procedure D) and
the deprotected compounds 14 may then be reacted further as
described above.
##STR00007##
General Procedures and Examples
[0274] The general synthetic schemes that were utilized to
construct the majority of compounds disclosed in this application
are described below in Schemes 1-9. These schemes are provided for
illustrative purposes only and are not to be construed as limiting
the scope of the invention.
##STR00008##
##STR00009##
##STR00010##
##STR00011##
##STR00012##
##STR00013##
##STR00014##
##STR00015##
##STR00016##
TABLE-US-00003 LIST OF GENERAL PROCEDURES General Procedure A
Nuclcophilic displacement of an aryl of heteroaryl halide with an
amine General Procedure B Buchwald-Hartwig reaction of an aryl of
heteroaryl halide with an amine General Procedure C Reaction of an
aryl or heteroaryl halide with a boronic acid or boronate ester
General Procedure D Acidic cleavage of a Boc-protected amine
General Procedure E Hydrogenation of a double bond General
Procedure F Formation of an amide from an acid chloride and an
amine or of a carbamate from a carbonochloridate and an amine
General Procedure G Chiral preparative HPLC separation of
stereoisomers General Procedure H Formation of a sulfonamide from a
sulfonyl chloride and an amine General Procedure I Formation of a
cyanamide from an amine from cyanogen bromide
[0275] The following examples are ordered according to the final
general procedure used in their preparation. The synthetic routes
to any novel intermediates are detailed by sequentially listing the
general procedure (letter codes) in parentheses after their name
with additional reactants or reagents as appropriate. A worked
example of this protocol is given below using Example #F.1.4 as a
non-limiting illustration. Example #F.1.4 is
8-((6-(1-acryloylpiperidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)am-
ino)-4,5-dihydro-1H-benzo[b]azepin-2(3H)-one, which was prepared
from
8-((6-(piperidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-dihy-
dro-1H-benzo[b]azepin-2(3H)-one using General Procedure F as
represented in Scheme A.
##STR00017##
[0276] The precursor to Example #F.1.4,
8-((6-(piperidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-dihy-
dro-1H-benzo[b]azepin-2(3H)-one, was prepared (as shown in Scheme
B) by initially reacting
6,8-dibromo-[1,2,4]triazolo[1,5-a]pyrazine, commercially available
from Ark Pharm, and
8-amino-4,5-dihydro-1H-benzo[b]azepin-2(3H)-one, commercially
available from AstaTech, following the conditions given in General
Procedure A, to give
8-((6-bromo-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-dihydro-1H-
-benzo[b]azepin-2(3H)-one, which is subsequently reacted with
tert-butyl
3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-
-carboxylate, commercially available from Anichem, using the
conditions given in General Procedure C to give tert-butyl
3-(8-((2-oxo-2,3,4,5-tetrahydro-1H-benzo
[b]azepin-8-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)-5,6-dihydropyri-
dine-1 (2H)-carboxylate, which is subsequently reacted with Pd/C
using the conditions provided in General Procedure E to give
tert-butyl
3-(8-((2-oxo-2,3,4,5-tetrahydro-1H-benzo[b]azepin-8-yl)amino)-[1,2,4]tria-
zolo[1,5-a]pyrazin-6-yl)piperidine-1-carboxylate, which is
subsequently reacted with TFA using the conditions given in General
Procedure D to give
8-((6-(piperidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-
-dihydro-1H-benzo[b]azepin-2(3H)-one. The reaction sequence to
synthesize the precursor to Example #F.1.4,
8-((6-(piperidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-dihy-
dro-1H-benzo[b]azepin-2(3H)-one, (detailed above) is consequently
translated in the preparations and examples section to:
8-((6-(piperidin-3-yl)-[1,2,4]triazolo[,5-a]pyrazin-8-yl)amino)-4,5-dihyd-
ro-1H-benzo[b]azepin-2(3H)-one (prepared using A from
6,8-dibromo-[1,2,4]triazolo[1,5-a]pyrazine [Ark Pharm] and
8-amino-4,5-dihydro-1H-benzo[b]azepin-2(3H)-one [AstaTech], C from
tert-butyl
3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-
-carboxylate [Anichem], E with Pd/C, D with TFA).
[0277] Hence the Example #F.1.4 would be written as: Example #F.1.4
was prepared from acryloyl chloride and
8-((6-(piperidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-dihy-
dro-1H-benzo[b]azepin-2(3H)-one (prepared using A from
6,8-dibromo-[1,2,4]triazolo[1,5-a]pyrazine [Ark Pharm] and
8-amino-4,5-dihydro-1H-benzo[b]azepin-2(3H)-one [AstaTech], C from
tert-butyl
3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-
-carboxylate [Anichem], E with Pd/C, D with TFA). In the tables
after a General Procedure, this is represented by having one
reactant in the title of the table and one in a separate column in
the same row as the product.
##STR00018## ##STR00019##
[0278] Analytical Methods
[0279] Analytical data was included within the procedures below, in
the illustrations of the general procedures, or in the tables of
examples. Unless otherwise stated, all .sup.1H NMR data were
collected on a Varian 400 MHz Mercury Plus, Inova, or 400-MR
instrument and chemical shifts are quoted in parts per million
(ppm). Optical rotation was determined using a Rudolph Research
Analytical Autopol IV automatic polarimeter at .lamda.=589 urn with
the samples dissolved in chloroform. LC/MS and HPLC data are
referenced to the table of LC/MS and HPLC conditions using the
lower case method letter provided in Table 1. Chiral SFC and HPLC
data are referenced to chiral SFC and HPLC conditions using the
numeric method letter provided in Table 2.
TABLE-US-00004 TABLE 1 LC/MS and HPLC methods Method Conditions a
LC/MS: The gradient was 5% B for 0.1 min, 5-100% B in 5.1 min with
a hold at 100% B for 0.5 min then 100-5% B in 0.3 min (2.0 mL/min
flow rate). Mobile phase A was 0.1% TFA in water, mobile phase B
was HPLC grade McCN. The column used for the chromatography was a
2.1 .times. 50 mm Phenomenex Luna Combi-HTS C8(2) column (5 .mu.m
particles) at a temperature of 65.degree. C. Detection methods are
diode array (DAD) and evaporative light scattering (ELSD) detection
under positive APCI ionization conditions. b LC/MS: The gradient
was 5% B for 0.1 min, 5-100% B in 2.5 min with a hold at 100% B for
0.3 min then 100-5% B in 0.1 min (2.0 mL/min flow rate). Mobile
phase A was 0.1% TFA in water, mobile phase B was HPLC grade MeCN.
The column used for the chromatography was a 2.1 .times. 50 mm
Phenomenex Luna Combi-HTS C8(2) column (5 .mu.m particles) at a
temperature of 55.degree. C. Detection methods arc diode array
(DAD) and evaporative light scattering (ELSD) detection under
positive APC1 ionization conditions. c LC/MS: The gradient was
10-90% B in 1.15 with a hold at 90% B for 0.40 min, 90-10% B in
0.01 min, and then hold at 10% B for 0.54 min (1.0 mL/min flow
rate). Mobile phase A was 0.0375% TFA in water, mobile phase B was
0.018% TFA in MeCN. The column used for the chromatography was a
2.1 .times. 30 mm Halo C18 column (2.7 .mu.m particles). Detection
methods are diode array (DAD) and positive/negative elecfrospray
ionization. d LC/MS: The gradient was 10-90% B in 3.4 min, 90-100%
B in 0.45 min. 100-1% B in 0.01 min, and then held at 1% B for 0.65
min (0.8 mL/min flow rate). Mobile phase A was 0.0375% TFA in
water, mobile phase B was 0.018% TFA in MeCN. The column used for
the chromatography was a 2.1 .times. 50 mm Venusil XBP-C18 column
(5 .mu.m particles). Detection methods arc diode array (DAD) and
evaporative light scattering (ELSD) detection as well as
positive/negative electrospray ionization. e LC/MS: The gradient
was 5% B for 0.1 min, 5-100% B in 5.1 min with a hold at 100% B for
0.5 min then 100-5% B in 0.3 min (2.0 mL/min flow rate). Mobile
phase A was 0.1% trifluoroacctic acid in water, mobile phase B was
HPLC grade MeCN. The column used for the chromatography was a 2.1
mm .times. 50 mm Phenomencx Luna Combi-HTS C8(2) column (5 .mu.m
particles) at a temperature of 55.degree. C. Detection methods are
diode array (DAD) and evaporative light scattering (ELSD) detection
under positive APCI ionization conditions. f LC/MS: The gradient
was 5-100% B in 3.4 min with a hold at 100% B for 0.45 min, 100-5%
B in 0.01 min, and then held at 5% B for 0.65 min (0.8 mL/min flow
rate). Mobile phase A was 10 mM NH.sub.4HCO.sub.3, mobile phase B
was HPLC grade MeCN. The column used for the chromatography is a
2.1 .times. 50 mm Xbridge Shield RPC18 column (5 .mu.m particles).
Detection methods arc diode array (DAD) and evaporative light
scattering (ELSD) detection as well as positive/negative
electrospray ionization. g LC/MS: The gradient was 5% B for 0.1
min, 5-100% B in 2.5 min with a hold at 100% B for 0.3 min then
100-5% B in 0.1 min (2.0 mL/min flow rate). Mobile phase A was 0.1%
TFA in water, mobile phase B was HPLC grade MeCN. The column used
for the chromatography was a 2.1 .times. 50 mm Phenomenex Luna
Combi-HTS C8(2) column (5 .mu.m particles) at a temperature of
65.degree. C. Detection methods are diode array (DAD) and
evaporative light scattering (ELSD) detection under positive APC1
ionization conditions. h LC/MS: The gradient was 5-60% B in 1.5 min
then 60-95% B to 2.5 min with a hold at 95% B for 1.2 min (1.3
mL/min flow rate). Mobile phase A was 0.1% formic acid in water,
mobile phase B was HPLC grade MeCN. The column used for the
chromatography is a 4.6 .times. 50 mm MAC-MOD Halo C8 column (2.7
.mu.m particles). Detection methods are diode array (DAD) and
evaporative light scattering (HLSD) detection as well as
positive/negative electrospray ionization. i HPLC: The gradient was
20-50% B in 12 min with a hold at 50% B for 2 min. then 50-100% B
in 0.2 min with a hold at 100% B for 2 min then 100-20% B in 0.2
min with a hold at 20% B for 1.6 min (1.0 mL/min flow rate). (25.0
mL/min flow rate). Mobile phase A was 0.75% TFA in water, mobile
phase B was MeCN. The column used for the chromatography was a 25
.times. 200 mm Phenomenex Luna C18 column (5 .mu.m particles).
Detection method is UV at wavelengths of 220 nm and 254 nm. j Prep
HPLC: The column is a Phenomenex Luna C18(2) 10 um 100.ANG. AXIA
column (250 mm x 21.2 mm). A gradient of MeCN (A) and 0.1% TFA in
water (B) is used, at a flow rate of 25 mL/min. A linear gradient
is used from about 5% of A to about 95% of A over about 10 minutes.
Detection method is UV at wave length of 220 nM and 254 nM. k HPLC:
The gradient was 2-60% B in 11.5 min then 60-95% B in 1 min with a
hold at 95% B for 2 min then 95-2% B in 0.5 min (25.0 mL/min flow
rate). (25.0 mL/min flow rate). Mobile phase A was 0.1% formic acid
in water, mobile phase B was MeCN. The column used for the
chromatography was a 50 .times. 100 mm Waters Atlantis T3 OBD
column (5 .mu.m particles). Detection method is UV at wavelength
range from 210 nm and 400 nm. l LC/MS (The gradient was 5-60% B in
1.5 min then 60-95% B to 2.5 min with a hold at 95% B for 1.2 min
(1.3 mL/min flow rate). Mobile phase A was 10 mM NH.sub.4OAc,
mobile phase B was HPLC grade MeCN. The column used for the
chromatography is a 4.6 .times. 50 mm MAC-MOD Halo C8 column (2.7
.mu.m particles). Detection methods arc diode array (DAD) and
evaporative light scattering (ELSD) detection as well as
positive/negative electrospray ionization.) m LC/MS: The gradient
was 5-95% B in 1.3 min then hold at 95% B for 1.5 min. back to 5% B
within 0.01 min (1.8 mL/min flow rate). Mobile phase A was 0.1%
NH.sub.4OAc in water, mobile phase B was HPLC grade MeCN. The
column used for the chromatography is a 4.6 .times. 50 mm XBridge
C18 (3.5 .mu.m particles). Detection methods arc diode array (DAD)
and evaporative light scattering (ELSD) detection as well as
positive/negative electrospray ionization. n LC/MS: The gradient
was 5-95% B in 1.4 min then hold at 95% B for 1.4 min, back to 5% B
within 0.01 min (1.8 mL/min flow rate). Mobile phase A was 0.1%
NH.sub.4OAc in water, mobile phase B was HPLC grade MeCN. The
column used for the chromatography is a 4.6 .times. 50 mm XBridge
C18 (3.5 .mu.m particles). Detection methods are diode array (DAD)
and evaporative light scattering (ELSD) detection as well as
positive/negative electrospray ionization. o LC/MS: The gradient
was 5-100% B in 1.3 min (2.0 mL/min flow rate). Mobile phase A was
0.1% TFA in water, mobile phase B was HPLC grade MeCN containing
0.1 % TFA. The column used for the chromatography is a 4.6 .times.
50 mm Sunfire C18 (3.5 .mu.m particles). Detection methods are
diode array (DAD) and evaporative light scattering (ELSD) detection
as well as positive/negative electrospray ionization. p LC/MS: The
gradient was 5-95% B in 1.3 min (1.8 mL/min flow rate). Mobile
phase A was 0.1% NH.sub.4OAC in water, mobile phase B was HPLC
grade MeCN. The column used for the chromatography is a 4.6 .times.
50 mm XBridge C18 (3.5 .mu.m particles). Detection methods are
diode array (DAD) and evaporative light scattering (ELSD) detection
as well as positive/negative electrospray ionization. q LC/MS: The
gradient was 5-95% B in 2.5 min (1.8 mL/min flow rate). Mobile
phase A was 0.1% NH.sub.4OAc in water, mobile phase B was HPLC
grade MeCN. The column used for the chromatography is a 4.6 .times.
50 mm Gemini-NX C18 (3.0 .mu.m particles). Detection methods arc
diode array (DAD) and evaporative light scattering (ELSD) detection
as well as positive/negative electrospray ionization. r LC/MS: The
gradient was 5-95% B in 1.4 min then hold at 95% B for 1.6 min,
back to 5% B within 0.01 min (1.8 mL/min flow rate). Mobile phase A
was 0.1% NH.sub.4OAc in water, mobile phase B was HPLC grade MeCN.
The column used for the chromatography is a 4.6 .times. 50 mm
XBridge C18 (3.5 .mu.m particles). Detection methods are diode
array (DAD) and evaporative light scattering (ELSD) detection as
well as positive/negative electrospray ionization. s LC/MS: The
gradient was 5-95% B in 1.2 min then hold at 95% B for 1.3 min,
back to 5% B within 0.01 min (1.8 mL/min flow rate). Mobile phase A
was 0.1% TFA in water, mobile phase B was HPLC grade MeCN (with
0.1% TFA). The column used for the chromatography is a 4.6 .times.
50 mm Sunfirc C18 (3.5 .mu.m particles). Detection methods are
diode array (DAD) and evaporative light scattering (ELSD) detection
as well as positive/negative electrospray ionization. t LC/MS: The
gradient was 10-100% B in 3.4 min with a hold at 100% B for 0.45
min, 100-10% B in 0.01 min, and then held at 10% B for 0.65 min
(0.8 mL/min flow rate). Mobile phase A was 0.0375%
CF.sub.3CO.sub.2H in water, mobile phase B was 0.018% CF3CO2H in
CH3CN. The column used for the chromatography was a 2.0 .times. 50
mm phenomenex Luna-C18 column (5 .mu.m particles). Detection
methods arc diode array (DAD) and evaporative light scattering
(ELSD) detection as well as positive electrospray ionization(MS). u
Prep HPLC: The column is a Luna C18 100*30 5u column. A gradient of
0.1% formic acid in water (A) and ACN (B) is used, at a flow rate
of 25 mL/min. A linear gradient is used from about 35% of B to 100%
of B over about 12 minutes. Detection method is UV at wave length
of 220 nM and 254 nM. v LC/MS: The gradient was 1-90% B in 3.4 min,
90-100% B in 0.45 min, 100-1% B in 0.01 min, and then held at 1% B
for 0.65 min (0.8 mL/min flow rate). Mobile phase A was 0.0375%
CF.sub.3CO.sub.2H in water, mobile phase B was 0.018%
CF.sub.3CO.sub.2H in CH.sub.3CN. The column used for the
chromatography was a 2.0 .times. 50 mm phenomenex Luna-C18 column
(5 .mu.m particles). Detection methods arc diode array (DAD) and
evaporative light scattering (ELSD) detection as well as positive
electrospray ionization(MS).) w LC/MS: The gradient was 10-90% B in
1.15 with a hold at 90% B for 0.4 min, 90-10% B in 0.01 min, and
then hold at 10% B for 0.54 min (1.0 mL/min flow rate). Mobile
phase A was 0.0375% trifluoroacetic acid in water, mobile phase B
was 0.018% trifluoroacetic acid in CH3CN. The column used for the
chromatography was a 2.1 .times. 30 mm Halo C18 column (2.7 .mu.m
particles). Detection methods are diode array (DAD) and positive
electrospray ionization(MS). x LC/MS: The gradient was 15-90% B in
3.4 min, 90-100% B in 0.45 min, 100- 15% B in 0.01 min, and then
held at 15% B for 0.65 min (0.8 mL/min flow rate). Mobile phase A
was 10 mM NH.sub.4HCO.sub.3, mobile phase B was HPLC grade ACN. The
column used for the chromatography is a 2.1 .times. 50 mm Xbridge
Shield RPC18 column (5 .mu.m particles). Detection methods are
diode array (DAD) and evaporative light scattering (ELSD) detection
as well as positive electrospray ionization(MS). y Prep HPLC: The
column used for the chromatography is a 19 .times. 50 mm Waters
Atlantis T3 OBD (5 .mu.m particles). Mobile phase A was 50 mM
ammonium acetate, mobile phase B was HPLC grade acetonitrile. The
gradient was a held at 16% B for 3 min, 16%-73% to 12.5 min and
then 73%-95.5% B to 13.5 min. Detection methods arc diode array
(DAD) and positive/negative ESI ionization. z Prep HPLC: The column
is a MAC-MOD: ACE C18 Prep, 5 um particle size 21.2 X 150 mm. A
gradient of 0.1% formic acid in water (A) and ACN (B) is used, at a
flow rate of 25 mL/min. A linear gradient is used from about 10% of
B to 95% of B over about 14 minutes. Detection method is UV at wave
length of 254 nM. aa Prep HPLC: The column is a MAC-MOD: ACE C18
Prep, 5 um panicle size 21.2 X 150 mm. A gradient of 0.1% formic
acid in water (A) and ACN (B) is used, at a flow rate of 25 mL/min.
The gradient used was 10% of B for 1.5 min, to 85% of B over 12.5
minutes, then 85-95% B to 13 minutes. Detection method is UV at
wave length of 254 nM. ab Prep HPLC: The column is a MAC-MOD: ACE
C18 Prep, 5 um particle size 21.2 X 150 mm. A gradient of 0.1%
formic acid in water (A) and ACN (B) is used, at a flow rate of 25
mL/min. The gradient used was 5% of B for 1.5 min, to 5-80% of B
over 12.5 minutes, then 80-95% B to 13 minutes. Detection method is
UV at wave length of 254 nM.
TABLE-US-00005 TABLE 2 Chiral SFC and HPLC methods Method
Conditions 1 Preparative SFC was performed on a THAR/Waters SFC 80
system running under SuperChrom software control. The preparative
SFC system was equipped with a CO.sub.2 pump, modifier pump,
automated back pressure regulator (ABPR), UV detector, injector,
and 6-position fraction collector. The mobile phase comprised of
supercritical CO.sub.2 supplied by a dcwar of bone-dry
non-certified CO.sub.2 pressurized to 800 psi with a modifier of
MeOH buffered with 0.05% NH.sub.4OH at a flow rate of 65 g/min. UV
detection was set to collect at a wavelength of 254 nm, the column
was at 40.degree. C. and the backpressure regulator was set to
maintain 100 bar. The sample was dissolved in MeOH. The mobile
phase was held isocratically at 25% MeOH (0.05% NH.sub.4OH):
CO.sub.2. The instrument was fitted with a ChiralCel OJ-H column
with dimensions 30 mm i.d. .times. 250 mm length with 5 .mu.m
particles. 2 Preparative SFC was performed on a THAR/Waters SFC 80
system running under SuperChrom software control. The preparative
SFC system was equipped with an 8-way preparative column switcher,
CO.sub.2 pump, modifier pump, automated hack pressure regulator
(ABPR), UV detector, and 6-position fraction collector. The mobile
phase comprised of supercritical CO.sub.2 supplied by a dewar of
bone-dry non- certified CO2 pressurized to 350 psi with a modifier
of MeOH buffered with 0.1% DEA at a flow rate of 70 g/min. UV
detection was set to collect at a wavelength of 220 nm. the column
was at ambient temperature, and the backpressure regulator was set
to maintain 100 bar. The sample was dissolved in MeOH at a
concentration of 75 mg/mL. The sample was loaded into the modifier
stream in 0.2 mL (15 mg) injections. The mobile phase was held
isocratically at 20% MeOH (0.1% DEA): CO.sub.2. Fraction collection
was time triggered. The instrument was fitted with a Regis Whelk-O
(S,S) column with dimensions 21 mm i.d. .times. 250 mm length with
5 .mu.m particles. 3 HPLC: The gradient was 25-37% B in 22.0 min
then 37-50% A in the next 7 min then held at 50% A for an extra 1
min. After the 1 min it is equilibrated back down to 25% for 3 min.
(20 mL/min flow rate). Mobile phase B was EtOH (200 proof), mobile
phase A was HPLC grade heptane with 0.20% DEA added. The column
used for the chromatography was a Daicel IF, 20 .times. 250 mm
column (5 .mu.m particles). Detection methods were UV (.lamda. =
298 nm) and optical rotation. 4 HPLC: The gradient was 17% B held
for 19 min then bumped up to 40% B for 5 min. After 5 min it is
equilibrated back down to 17% for 3 min (20 mL/min flow rate).
Mobile phase B was HPLC grade isopropanol, mobile phase A was HPLC
grade heptane with 0.20% DBA added. The column used for the
chromatography was a Daicel IA, 20 .times. 250 mm column (5 .mu.m
particles). Detection methods were UV (.lamda. = 322 nm) and
optical rotation. 5 HPLC: The gradient was 15-29% A in 26 min then
step to 60% A for 4 min (20 mJL/min How rate). Mobile phase A was
EtOH (200 proof), mobile phase B was HPLC grade heptane no
modifier. The chromatography used a Daicel IB, 20 .times. 250 mm
column (5 .mu.m particles). 6 HPLC: Isocratic 17% A for 19 min then
step to 40% A for 6 min (20 mL/min flow rate). Mobile phase A was
EtOH (200 proof), mobile phase B was HPLC grade heptane with 0.1%
DEA added. The chromatography used a Daicel IA, 21 .times. 250 mm
column (5 .mu.m particles). 7 HPLC: The gradient was 25-37% A in 22
min then 37-50% in 7 min (20 mL/min flow rate). Mobile phase A was
EtOH (200 proof), mobile phase B was HPLC grade heptane with 0.1%
DEA added. The chromatography used a Daicel IF, 20 .times. 250 mm
column (5 .mu.m particles). 8 HPLC was performed using the
following step gradient: the gradient was 25 to 35% B in 16 min
then stepped to 65% B in .05 min, then 65-80% B in the next 6.95
min. It is equilibrated back to 25% for 4 min. (20 mL/min flow
rate). Mobile phase B was EtOH (200 proof), mobile phase A was HPLC
grade heptane with 0.20% DEA added. The chromatography used a
Daicel IC, 20 .times. 250 mm column (5 .mu.m particles). 9 HPLC:
The gradient was 36-45% A in 19.5 min then 65-80% A in 5.4 min (20
mL/min flow rate). Mobile phase A was HPLC grade EtOAc, mobile
phase B was HPLC grade heptane with 0.2% DBA added. The
chromatography used a Daicel ID, 21 .times. 250 nun column (5 .mu.m
particles). 10 HPLC: Isocratic 31% A for 32 min (20 mL/min flow
rate). Mobile phase A was HPLC grade isopropanol, mobile phase B
was HPLC grade heptane with no modifier. The chromatography used a
Daicel IB, 20 .times. 250 mm column (5 .mu.m particles) 11 HPLC:
Isocratic 30% A. (1 mL/min flow rate). Mobile phase A was HPLC
grade EtOH with 1% DEA added, mobile phase B was HPLC grade
n-hexane with 0.1% DEA added. The column used for the
chromatography was a AD-H, 4.6 .times. 250 mm column (5 .mu.m
particles). Detection methods were UV (.lamda. = 214, 254 nm). 12
HPLC: Isocratic 30% A. (3 mL/min flow rate). Mobile phase A was
HPLC grade EtOH with 0.1% DEA added, mobile phase B was HPLC grade
n-hexane with 0.1% DEA added. The column used for the
chromatography was a OZ-H, 4.6 .times. 250 mm column (5 .mu.m
particles). 13 HPLC: Isocratic 20% A. (3 mL/min flow rate). Mobile
phase A was HPLC grade MeOH with 0.1% DEA added, mobile phase B was
HPLC grade n-hexane with 0.1% DEA added. The column used for the
chromatography was a AS-H, 4.6 .times. 250 mm column (5 .mu.m
particles). 14 HPLC: Isocratic 50% A. (1 mL/min flow rate). Mobile
phase A was HPLC grade EtOH with 0.1% DEA added, mobile phase B was
HPLC grade n-hexane with 0.1% DEA added. The column used for the
chromatography was a 1A. 4.6 .times. 250 mm column (5 .mu.m
particles). Detection methods were UV (.lamda. = 214, 254 nm). 15
HPLC: Isocratic 15% A. (3 mL/min flow rate). Mobile phase A was
HPLC grade MeOH with 0.1% DEA added, mobile phase B was HPLC grade
n-hexane with 0.1 % DEA added. The column used for the
chromatography was a OJ-H. 4.6 .times. 250 mm column (5 .mu.m
particles). 16 HPLC 2-Dimensional purification: Dim 1: 35% A for 20
min (20 mL/min flow rate). Mobile phase A was EtOH (200 proof),
mobile phase B was HPLC grade heptane with 0.2% diethylaminc added.
The chromatography used a Daicel IC 20 .times. 250 mm column (5
.mu.m particles). Dim 2: 30% A for 14 min (20 mL/min flow rate).
Mobile phase A was EtOH (200 proof), mobile phase B was HPLC grade
heptane with 0.2% diethylamine added. The chromatography used a
Cel-4, 21 .times. 250 mm column (5 .mu.m particles) from
Phenomenex. 17 Preparative SFC was performed on a THAR/Waters SFC
80 system running under SuperChrom software control. The
preparative SFC system was equipped with a CO.sub.2 pump, modifier
pump, automated back pressure regulator (ABPR), UV detector,
injector, and 6-position fraction collector. The mobile phase
comprised of supercritical CO.sub.2 supplied by a dewar of bone-dry
non-certified CO.sub.2 pressurized to 800 psi with a modifier of
MeOH buffered with 0.1% NH.sub.3 in water at a flow rate of 60
g/min. UV detection was set to collect at a wavelength of 254 nm,
the backpressure regulator was set to maintain 100 bar. The sample
was dissolved in MeOH. The mobile phase was held isocratically at
20% MeOH (0.1% NH.sub.3 in water): CO.sub.2. The instrument was
fitted with a ChiralCel OJ 10 mm coluum with dimensions 3.0 cm i.d.
.times. 50 cm length 18 Preparative SFC was performed on a
THAR/Waters SFC 80 system running under SuperChrom software
control. The preparative SFC system was equipped with a CO.sub.2
pump, modifier pump, automated back pressure regulator (ABPR), UV
detector, injector, and 6-position fraction collector. The mobile
phase comprised of supercritical CO.sub.2 supplied by a dewar of
bone-dry non-certified CO.sub.2 pressurized to 800 psi with a
modifier of IPA buffered with 0.1 % NH.sub.3 in H.sub.2O at a flow
rate of 60 g/min. UV detection was set to collect at a wavelength
of 254 nm, the backpressure regulator was set to maintain 100 bar.
The sample was dissolved in IPA. The mobile phase was held
isocratically at 20% IPA (0.1% NH.sub.3 in water): CO.sub.2. The
instrument was fitted with a ChiralCel OJ 10 mm column with
dimensions 3.0 cm i.d. .times. 50 cm length 19 Preparative SFC was
performed on a THAR/Waters SFC 80 system running under SuperChrom
software control. The preparative SFC system was equipped with a
CO.sub.2 pump, modifier pump, automated back pressure regulator
(ABPR), UV detector, injector, and 6-position fraction collector.
The mobile phase comprised of supercritical CO.sub.2 supplied by a
dewar of bone-dry non-certified CO.sub.2 pressurized to 800 psi
with a modifier of IPA buffered with 0.1 % NH.sub.4OH at a flow
rate of 55 g/min. UV detection was set to collect at a wavelength
of 220 nm, the backpressure regulator was set to maintain 100 bar.
The sample was dissolved in IPA. The mobile phase was held
isocratically at 70% IPA (0.1% NH.sub.3 in water): CO.sub.2. The
instrument was fitted with a Chiralpak AD-H 5 .mu.m , 3.0 cm id
.times. 25 cm length
Purification Methods
[0280] For examples without detailed procedures, the compounds may
be purified by any technique or combination of techniques known to
one skilled in the art. Some examples that are not limiting include
flash chromatography with a solid phase (e.g. silica gel, alumina,
etc.) and a solvent (or combination of solvents) that elutes the
desired compounds (e.g. heptane, EtOAc, DCM, MeOH, MeCN, water,
etc.); preparatory TLC with a solid phase (e.g. silica gel, alumina
etc.) and a solvent (or combination of solvents) that elutes the
desired compounds (e.g. heptane, EtOAc, DCM, MeOH, MeCN, water,
etc.); reverse phase HPLC (see Table 1 for some non-limiting
conditions); recrystallization from an appropriate solvent or
combination of solvents (e.g. MeOH, EtOH, i-PrOH, EtOAc, toluene,
etc.) or combination of solvents (e.g. EtOAc/heptane, EtOAc/MeOH,
etc.); chiral HPLC with a solid phase and an appropriate solvent
(see Table 2 for some non-limiting conditions) to elute the desired
compound; chiral SFC with a solid phase and CO.sub.2 with an
appropriate modifier (e.g. MeOH, EtOH, i-PrOH with or without
additional modifier such as DEA, TFA, etc.); precipitation from a
combination of solvents (e.g. DMF/water, DMSO/DCM, EtOAc/heptane,
etc.); trituration with an appropriate solvent (e.g. EtOAc, DCM,
MeCN, MeOH, EtOH, i-PrOH, etc.); extractions by dissolving a
compound in a liquid and washing with an appropriately immiscible
liquid (e.g. DCM/water, EtOAc/water, DCM/saturated aqueous
NaHCO.sub.3, EtOAc/saturated aqueous NaHCO.sub.3, DCM/10% aqueous
HCl, EtOAc/10% aqueous HCl, etc.); distillation (e.g. simple,
fractional, Kugelrohr, etc.); gas chromatography using an
appropriate temperature, carrier gas and flow rate; sublimation at
an appropriate temperature and pressure; filtration through a media
(e.g. Florosil.RTM., alumina, Celite.RTM., silica gel, etc.) with a
solvent (e.g. heptane, hexanes, EtOAc, DCM, MeOH, etc.) or
combination of solvents; salt formation with solid support (resin
based, e.g. ion exchange) or without. Compounds of interest may be
isolated as a salt without the use of a specific salt formation
purification method. For example, on occasions where purification
is accomplished with reverse phase HPLC with an aqueous TFA buffer,
the TFA salt may be isolated. Some descriptions of these techniques
can be found in the following references: Gordon, A. J. and Ford,
R. A., "The Chemist's Companion", 1972; Palleros, D. R.
"Experimental Organic Chemistry", 2000; Still, W. C., Kahn and M.
Mitra, A. J. Org. Chem. 1978, 43, 2923; Yan, B. "Analysis and
Purification Methods in Combinatorial Chemistry", 2003; Harwood, L.
M., Moody, C. J. and Percy, J. M. "Experimental Organic Chemistry:
Standard and Microscale, 2.sup.nd Edition", 1999; Stichlmair, J. G.
and Fair, J. R. "Distillation; Principles and Practices", 1998;
Beesley, T. E. and Scott, R. P. W. "Chiral Chromatography", 1999;
Landgrebe, J. A. "Theory and Practice in the Organic Laboratory,
4.sup.th Ed.", 1993; Skoog, D. A. and Leary, J. J. "Principles of
Instrumental Analysis, 4.sup.th Ed.", 1992; G. Subramanian, "Chiral
Separation Techniques, 3.sup.rd Edition", 2007; Y. Kazakevich, R.
Lobrutto, "HPLC for Pharmaceutical Scientists", 2007. Intermediates
and final compounds prepared via the General Procedures listed
below can be optionally purified using one or more of the
Purification Methods described above.
PREPARATIONS AND EXAMPLES
[0281] All starting materials are commercially available from
Sigma-Aldrich (including Fluka, Aldrich Market Select, and
Discovery CPR) unless otherwise noted after the chemical name.
Reagent/reactant names given are as named on the commercial bottle
or as generated by IUPAC conventions, CambridgeSoft.RTM. ChemDraw
Ultra 12.0, CambridgeSoft.RTM. Chemistry E-Notebook 11, or AutoNom
2000. The general synthetic methods used in each General Procedure
follow and include an illustration of a compound that was
synthesized using the designated General Procedure. None of the
specific conditions and reagents noted herein are to be construed
as limiting the scope of the invention and are provided for
illustrative purposes only. Compounds designated as salts (e.g.
hydrochloride, trifluoroacetate) may contain more than one molar
equivalent of the salt or may contain the acid as an excipient.
Compounds of the invention where the absolute stereochemistry has
been determined by the use of a commercially available
enantiomerically pure starting material or a stereochemically
defined intermediate or by X-ray diffraction are denoted by an
asterisk after the example number. Otherwise the absolute
stereochemistry is unknown and assigned randomly as drawn.
Preparation #1:
6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-amine
##STR00020##
[0282] Step A. methyl 3-nitro-1H-pyrazole-5-carboxylate
##STR00021##
[0284] To a solution of 3-nitro-1H-pyrazole-5-carboxylic acid
(69.75 g, 444 mmol) (ArkPharm) in MeOH (1 L) was added thionyl
chloride (84 mL, 1154 mmol) at 0.degree. C. The mixture was stirred
for about 20 min at about 0.degree. C. then heated to reflux for
about 2 h. The resulting solution was concentrated under reduced
pressure to give methyl 3-nitro-1H-pyrazole-5-carboxylate (61.25 g,
81%): LC/MS (Table 1, Method 1) R.sub.t=1.42 min.; MS m/z: 169
(M+H).sup.+.
Step B. methyl
1-(2-bromoethyl)-3-nitro-1H-pyrazole-5-carboxylate
##STR00022##
[0286] A 3 L 3-necked flask fitted with reflux condenser and
thermocoupler was charged with methyl
3-nitro-1H-pyrazole-5-carboxylate (75.5 g, 441 mmol) and DMF (735
mL). Cesium carbonate (173 g, 529 mmol) was added portionwise and
the reaction was heated to about 98.degree. C. for 5 min, then
cooled to ambient temperature for about 30 min. The reaction was
cooled in an ice bath to about 0.degree. C. before the addition of
1,2-dibromoethane (380 mL, 4412 mmol). The reaction was stirred,
warming to ambient temperature, for about 5 h. The reaction mixture
was quenched with the addition of an aqueous solution of potassium
phosphate monobasic (120 g in 1 L). The resulting solution was
extracted with EtOAc (3.times.300 mL). The combined organic portion
was dried over MgSO4, filtered and concentrated under reduced
pressure to afford methyl
1-(2-bromoethyl)-3-nitro-H-pyrazole-5-carboxylate (120 g, 92%):
LC/MS (Table 1, Method 1) R.sub.t=2.10 min.; MS m/z: 278, 280
(M+H).sup.+.
Step C. (1-(2-bromoethyl)-3-nitro-1H-pyrazol-5-yl)methanol
##STR00023##
[0288] A 3 L flask fitted with addition funnel and chilled in an
ice bath was charged with lithium tetrahydroborate (259 mL, 518
mmol) (2N in THF) and THF (252 mL). The reaction mixture was cooled
to about 0.degree. C. before the dropwise addition of a solution of
methyl 1-(2-bromoethyl)-3-nitro-1H-pyrazole-5-carboxylate (72 g,
259 mmol) in THF (126 mL). The reaction stirred for about 2 h at
ambient temperature. The reaction mixture was quenched with the
addition of aqueous saturated NaCl (400 mL). The resulting mixture
was extracted with EtOAc (3.times.400 mL). The combined organic
portion was dried over MgSO4, filtered, and concentrated under
reduced pressure to afford
(1-(2-bromoethyl)-3-nitro-H-pyrazol-5-yl)methanol (56.1 g, 87%):
LC/MS (Table 1, Method 1) R.sub.t=1.52 min.; MS m/z: 250, 252
(M+H).sup.+.
Step D. 2-nitro-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine
##STR00024##
[0290] A 2 L flask was charged with
(1-(2-bromoethyl)-3-nitro-1H-pyrazol-5-yl)methanol (56 g, 190 mmol)
and dissolved in DMA (747 mL). The reaction was heated to about
140.degree. C. for about 5 h. The reaction cooled to ambient
temperature and the solvent was concentrated under reduced
pressure. The resulting residue was partitioned between EtOAc (500
mL) and aqueous saturated NaHCO.sub.3 (150 mL). The aqueous portion
was extracted with EtOAc (3.times.400 mL). The combined organic
portion was dried over MgSO.sub.4, filtered, and concentrated under
reduced pressure. 200 mL of Et.sub.2O was added to the resulting
residue and the solid was collected via filtration to afford
2-nitro-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine (14 g, 43.5%).
The remaining filtrate was concentrated under reduced pressure and
purified using silicagel chromatography to afford
2-nitro-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine (6.5 g, 38.4
mmol, 20.19% yield):): LC/MS (Table 1, Method 1) R.sub.t=1.31 min.;
MS m/z: 170 (M+H).sup.+.
Step E. 6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-amine
##STR00025##
[0292] A flask was charged with 10% palladium on carbon (2.64 g,
2.483 mmol). The flask was evacuated and put under nitrogen
atmosphere before the addition of MeOH (100 mL) and
2-nitro-6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazine (14 g, 83 mmol)
in EtOAc (300 mL). The reaction was evacuated and purged with
hydrogen three times. The reaction stirred at ambient temperature
for about 16 h. The catalyst was filtered off through a pad of
Celite.RTM. and the compound was washed with about 300 mL of EtOAc.
The solvent was concentrated under reduced pressure to give,
6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-amine (10.9 g, 95%):
LC/MS (Table 1, Method 1) R.sub.t=0.61 min.; MS m/z: 140
(M+H).sup.+.
Preparation #2: tert-butyl
3-(8-((tert-butoxycarbonyl)(6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-y-
l)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)azetidine-1-carboxylate
##STR00026##
[0293] Step A:
N-(6-bromo-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5,-
1-c][1,4]oxazin-2-amine
##STR00027##
[0295] To a microwave reaction vial were added tert-butyl
6,8-dibromo-[1,2,4]triazolo[1,5-a]pyridine (1.0 g, 3.6 mmol,
ArkPharm), 6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-amine (0.503
g, 3.61 mmol, Preparation #1), 1,4-dioxane (12 mL),
Cs.sub.2CO.sub.3 (2.353 g, 7.22 mmol), Xantphos (0.104 g, 0.181
mmol) and Pd.sub.2(dba).sub.3 (0.165 g, 0.181 mmol), The reaction
vial was flushed with nitrogen, capped, stirred and heated to about
120.degree. C. in a Biotage microwave reactor for about 3 h. The
reaction was diluted with DCM (80 mL) and water (50 mL). The
organic layer was separated, washed with water (50 mL), brine (50
mL), and dried over Na.sub.2SO.sub.4. The organic layer was
filtered and concentrated under reduced pressure The crude product
was purified via silica chromatography eluting with 5% MeOH in DCM
to afford
N-(6-bromo-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5,-
1-c][1,4]oxazin-2-amine (0.80 g, 43%) as a yellow solid: LC/MS
(Table 1, Method p) R.sub.t=1.59 min.; MS m/z: 335/337
(M+H).sup.+.
Step B: tert-butyl
(6-bromo-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-dihydro-4H-pyrazolo
[5,1-c][1,4]oxazin-2-yl)carbamate
##STR00028##
[0297] A mixture of
N-(6-bromo-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5,-
1-c][1,4]oxazin-2-amine (0.80 g, 1.5 mmol), BOC.sub.2O (1.08 mL,
4.65 mmol), TEA (0.649 mL, 4.65 mmol) and DMAP (0.190 g, 1.55 mmol)
in DCM, (60 mL) was stirred at rt overnight. The organic layer was
washed with saturated NH.sub.4Cl (3.times.50 mL). The organic layer
was dried with Na.sub.2SO.sub.4, filtered and concentrated. The
product was purified via silica chromatography eluting with
EtOAc:petroleum ether (2:1) to afford tert-butyl
(6-bromo-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-dihydro-4H-pyrazolo[5,1--
c][1,4]oxazin-2-yl)carbamate (0.53 g, 77%) as white solid: LC/MS
(Table 1, Method n) R.sub.t=1.73 min.; MS m/z: 435/437
(M+H).sup.+.
Step C: tert-butyl
3-(8-((tert-butoxycarbonyl)(6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-y-
l)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)azetidine-1-carboxylate
##STR00029##
[0299] To a mixture of zinc (0.16 g, 2.5 mmol) in degassed DMA (3
mL) under nitrogen was added trimethylsilyl chloride (0.032 mL,
0.25 mmol) and 1,2-dibromoethane (0.032 g, 0.17 mmol). The mixture
was stirred for about 15 min, and tert-butyl
3-iodoazetidine-1-carboxylate (0.35 g, 1.2 mmol) was added via
syringe. The resulting mixture was stirred at rt for about 1.5 h to
form the (1-(tert-butoxycarbonyl)azetidin-3-yl) zinc (II) iodide.
tert-Butyl
(6-bromo-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-dihydro-4H-pyrazolo[5,1--
c][1,4]oxazin-2-yl)carbamate (0.11 g, 0.25 mmol) was dissolved in
DMA (5 mL), and degassed for about 5 min followed by the addition
of PdCl2(dppf) (0.013 g, 0.017 mmol) and copper(I)iodide (0.056 g,
0.30 mmol) and then the addition of the previously generated
solution of (1-(tert-butoxycarbonyl)azetidin-3-yl) zinc(II) iodide.
The reaction was heated to about 80.degree. C. for about 2 h. The
reaction mixture was diluted with EtOAc (40 mL) and filtered
through a nylon filter. Water (40 mL) was added, the layers were
separated, the organic phase was washed with brine (3.times.40
mL)), and then dried over anhydrous Na.sub.2SO.sub.4. The solution
was concentrated and the residue was purified by prep-TLC using DCM
and MeOH (40:1) and tert-butyl
3-(8-((tert-butoxycarbonyl)(6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-y-
l)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)azetidine-1-carboxylate
(0.07 g, 41.5%) was obtained as a brown solid: LC/MS (Table 1,
Method q) R.sub.t=1.81 min.; MS m/z: 512 (M+H).sup.+.
Preparation #3: 1-(4-amino-1H-pyrazol-1-yl)-2-methylpropan-2-ol
##STR00030##
[0300] Step A: 2-methyl-1-(4-nitro-1H-pyrazol-1-yl)propan-2-ol
##STR00031##
[0302] A round bottom flask was charged with 4-nitro-1H-pyrazole (4
g, 35.4 mmol), 2,2-dimethyloxirane (5.1 g, 70 mmol), and
Cs.sub.2CO.sub.3 (23 g, 70 mmol). The reaction was heated to about
90.degree. C. for about 12 h, cooled to ambient temperature,
filtered, and the filtrate was concentrated under reduced pressure.
The crude material was purified via silica chromatography eluting
with EtOAc:petroleum ether (10:1 to 3:1) to afford
2-methyl-1-(4-nitro-1H-pyrazol-1-yl)propan-2-ol (4 g, 61% yield) as
a yellow oil. LC/MS (Table 1, Method w) R.sub.t=0.976 min.; MS m/z:
182 (M+H).sup.+.
Step B: 1-(4-amino-1H-pyrazol-1-yl)-2-methylpropan-2-ol
##STR00032##
[0304] A round bottom flask was charged with
2-methyl-1-(4-nitro-1H-pyrazol-1-yl)propan-2-ol (1 g, 5.40 mmol)
and Raney Nickel (1 g) in THF (40 mL). The reaction mixture stirred
at about 20.degree. C. for about 12 h, under an atmosphere of
hydrogen. The reaction mixture was filtered through a pad of
Celite.RTM. and concentrated under reduced pressure to afford
I-(4-amino-1H-pyrazol-1-yl)-2-methylpropan-2-ol (0.8 g, 95% yield)
as a white solid. LC/MS (Table 1, Method w) R.sub.t=0.155 min.; MS
m/z: 156 (M+H).sup.+.
Preparation #4: 4-(4-amino-1H-pyrazol-1-yl)-2-methylbutan-2-ol
##STR00033##
[0305] Step A: 3-hydroxy-3-methylbutyl methanesulfonate
##STR00034##
[0307] A round bottom flask was charged with
3-methylbutane-1,3-diol (10 g, 96 mmol) and TEA (20 mL, 144 mmol)
in DCM (80 mL). A solution of methanesulfonyl chloride (12 g, 106
mmol) in DCM (50 mL) was added dropwise to the reaction mixture at
about 0.degree. C. The resulting mixture was stirred at about
0.degree. C. for 4 h. The reaction was diluted with saturated aq.
sodium bicarbonate (100 mL). The organic portion was dried over
anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure to give 3-hydroxy-3-methylbutyl methanesulfonate
(12 g, 66% yield) as a yellow oil. .sup.1H NMR (400 MHz,
CHLOROFORM-d) 6=4.40 (t, J=7.1 Hz, 2H), 3.01 (s, 3H), 1.94 (t,
J=7.1 Hz, 2H), 1.81 (s, 1H), 1.27 (s, 6H).
Step B: 2-methyl-4-(4-nitro-1H-pyrazol-1-yl)butan-2-ol
##STR00035##
[0309] A round bottom flask was charged with
3-hydroxy-3-methylbutyl methanesulfonate (12 g, 66 mmol),
K.sub.2CO.sub.3 (7.3 g, 53.1 mmol) and KI (4.4 g, 26.5 mmol) ACN
(250 mL). 4-nitro-1H-pyrazole (3 g, 26.5 mmol) was added to the
reaction mixture at about 15.degree. C. The reaction was then
heated to about 90.degree. C. for about 12 h. The reaction mixture
was cooled to ambient temperature, diluted with water (100 mL) and
extracted with EtOAc (3.times.200 mL). The combined organic portion
was dried over anhydrous Na.sub.2SO.sub.4, filtered, and
concentrated under reduced pressure. The crude material was
purified via silica chromatography eluting with: petroleum
ether:EtOAc (30:1 to 1:1) to afford
2-methyl-4-(4-nitro-1H-pyrazol-1-yl)butan-2-ol (4.7 g, 89% yield)
as a yellow oil. LC/MS (Table 1, Method w) R.sub.t=0.807 min.; MS
m/z: 200 (M+H).sup.+.
Step C: 4-(4-amino-1H-pyrazol-1-yl)-2-methylbutan-2-ol
##STR00036##
[0311] To a solution of
2-methyl-4-(4-nitro-1H-pyrazol-1-yl)butan-2-ol (0.5 g, 2.5 mmol) in
THF (50 mL) was added Raney nickel (1 g, 17.04 mmol) at about
15.degree. C. The reaction mixture stirred at about 15.degree. C.
for about 12 h, under an atmosphere of hydrogen. The reaction
mixture was filtered through a pad of Celite.RTM. and concentrated
under reduced pressure to afford
4-(4-amino-1H-pyrazol-1-yl)-2-methylbutan-2-ol (0.36 g, 80% yield)
as a light pink solid. LC/MS (Table 1, Method w) R.sub.t=0.162
min.; MS m/z: 170 (M+H).sup.+.
Preparation #5:
1-(2,2,6,6-tetramethyltetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-amine
##STR00037##
[0312] Step A: 2,2,6,6-tetramethyldihydro-2H-pyran-4(3H)-one
##STR00038##
[0314] A round bottom flask was charged with 2,6-dimethyl
hepta-2,5-dien-4-one (10 g, 72.4 mmol) and 1 M HCl (100 mL, 100
mmol). The reaction was stirred at about 40.degree. C. for about 4
days. The reaction was extracted with DCM (3.times.100 mL). The
organic portion was dried over anhydrous Na.sub.2SO.sub.4,
filtered, and concentrated under reduced pressure to give
2,2,6,6-tetramethyldihydro-2H-pyran-4(3H)-one (10 g, 62% yield) as
a yellow oil. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=2.15-2.10
(m, 2H), 1.91-1.87 (m, 2H), 1.31 (s, 6H), 1.27 (s, 6H).
Step B: 2,2,6,6-tetramethyltetrahydro-2H-pyran-4-ol
##STR00039##
[0316] To a solution of
2,2,6,6-tetramethyldihydro-2H-pyran-4(3H)-one (2 g, 12.8 mmol) in
MeOH (50 mL) was added portionwise NaBH.sub.4 (0.97 g, 25.6 mmol)
at about 0.degree. C. The reaction stirred at about 0.degree. C.
for about 2 h. The reaction was diluted with saturated aq.
NH.sub.4Cl (50 mL) and extracted with DCM (2.times.50 mL). The
organic portion was dried over anhydrous Na.sub.2SO.sub.4,
filtered, and concentrated under reduced pressure to give
2,2,6,6-tetramethyltetrahydro-2H-pyran-4-ol (1.4 g, 48% yield) as a
white wax. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=4.1-4.05 (m,
1H), 1.92-1.87 (m, 2H), 1.71-1.68 (m, 2H), 1.25 (s, 6H), 1.23 (s,
6H).
Step C: 2,2,6,6-tetramethyltetrahydro-2H-pyran-4-yl
methanesulfonate
##STR00040##
[0318] A round bottom flask was charged with
2,2,6,6-tetramethyltetrahydro-2H-pyran-4-ol (5.5 g, 34.8 mmol) and
TEA (7.0 g, 69 mmol) in DCM (110 mL). Methanesulfonyl chloride (5.9
g, 52.1 mmol) was slowly added to the reaction mixture at about
0.degree. C. over 20 minutes. The reaction warmed to about
20.degree. C. over 30 minutes and stirred at about 20.degree. C.
for about 2 h. The reaction was diluted with water (100 mL) and
extracted with DCM (3.times.100 mL). The organic portion was dried
over anhydrous Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure to give
2,2,6,6-tetramethyltetrahydro-2H-pyran-4-yl methanesulfonate (8 g,
crude) as a yellow oil. .sup.1H NMR (400 MHz, CHLOROFORM-d)
.delta.=5.1-5.09 (m, 1H), 3.03 (s, 3H), 2.1-2.07 (d, 2H), 1.59-1.54
(d, 2H), 1.30 (s, 6H), 1.25 (s, 6H).
Step D:
4-nitro-1-(2,2,6,6-tetramethyltetrahydro-2H-pyran-4-yl)-1H-pyrazol-
e
##STR00041##
[0320] A round bottom flask was charged with 4-nitro-1H-pyrazole
(1.2 g, 10.61 mmol), 2,2,6,6-tetramethyltetrahydro-2H-pyran-4-yl
methanesulfonate (7.5 g, 31 mmol), and Cs.sub.2CO.sub.3 (6.9 g,
21.2 mmol) in DMF (30 mL). The reaction was heated to about
130.degree. C. for about 12 h. The reaction was cooled to ambient
temperature, diluted with water (100 mL) and extracted with EtOAc
(3.times.100 mL). The organic portion was dried over anhydrous
Na.sub.2SO.sub.4, filtered, and concentrated under reduced
pressure. The crude material was purified via siliacgel
chromatography eluting with petroleum ether/EtOAc (20:1 to 5:1) to
give
4-nitro-1-(2,2,6,6-tetramethyltetrahydro-2H-pyran-4-yl)-1H-pyrazole
(0.6 g, 21% yield) as white solid. LC/MS (Table 1, Method w)
R.sub.t=1.37 min.; MS m/z: 254 (M+H).sup.+.
Step E:
1-(2,2,6,6-tetramethyltetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-amine
##STR00042##
[0322] To a solution of of
4-nitro-1-(2,2,6,6-tetramethyltetrahydro-2H-pyran-4-yl)-1H-pyrazole
(0.3 g, 1.18 mmol) in THF (20 mL) was added Raney nickel (1 g,
17.04 mmol). The reaction mixture stirred at about 20.degree. C.
for about 12 h, under an atmosphere of hydrogen. The reaction
mixture was filtered through a pad of Celite.RTM. and concentrated
under reduced pressure to afford
1-(2,2,6,6-tetramethyltetrahydro-2H-pyran-4-yl)-1H-pyrazol-4-amine
(0.25 g, 85% yield) as a white solid. LC/MS (Table 1, Method w)
R.sub.t=0.729 min.; MS m/z: 224 (M+H).sup.+.
Preparation #6: (trans)-4-(4-amino-1H-pyrazol-1-yl)cyclohexanol and
(cis)-4-(4-amino-1H-pyrazol-1-yl)cyclohexanol
##STR00043##
[0323] Step A: 4-hydroxycyclohexyl methanesulfonate
##STR00044##
[0325] A round bottom flask was charged with cyclohexane-1,4-diol
(10 g, 86 mmol) and TEA (8.7 g, 86 mmol) in THF (200 mL).
Methanesulfonyl chloride (3.9 g, 34 mmol) was slowly added to the
reaction mixture at about 0.degree. C. over 20 minutes. The
reaction warmed to about 20.degree. stirred for about 2 h. The
reaction was diluted with water (150 mL) and extracted with EtOAc
(3.times.150 mL). The organic portion was dried over anhydrous
Na.sub.2SO.sub.4, filtered, and concentrated under reduced pressure
to give 4-hydroxycyclohexyl methanesulfonate (6 g, 32% yield) as a
yellow oil. .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=4.8-4.71
(m, 1H), 3.79-3.73 (m, 1H), 3.02 (s, 3H), 2.1-1.97 (m, 4H),
1.6-1.46 (m, 4H).
Step B: (trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexanol and
(cis)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexanol
##STR00045##
[0327] A round bottom flask was charged with 4-nitro-1H-pyrazole (2
g, 17.6 mmol), 4-hydroxycyclohexyl methanesulfonate (6 g, 30.9
mmol), and Cs.sub.2CO.sub.3 (11.5 g, 35.4 mmol) in DMF (60 mL). The
reaction was heated to about 130.degree. C. for about 12 h. The
reaction was cooled to ambient temperature and concentrated under
reduced pressure. The crude material was purified via siliacgel
chromatography eluting with petroleum ether/EtOAc (10:1 to 1:1) to
give (trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexanol (0.3 g, 8%
yield). LC/MS (Table 1, Method w) R.sub.t=1.08 min.; MS m/z: 211
(M+H).sup.+, and (cis)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexanol (0.3
g, 8% yield).). LC/MS (Table 1, Method w) R.sub.t=1.09 min.; MS
m/z: 211 (M+H).sup.+
Step C: (trans)-4-(4-amino-1H-pyrazol-1-yl)cyclohexanol
##STR00046##
[0329] To a solution of
(trans)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexanol (0.15 g, 0.71 mmol)
in THF (15 mL) was added Raney nickel (0.3 g). The reaction mixture
stirred at about 20.degree. C. for about 3 h, under an atmosphere
of hydrogen. The reaction mixture was filtered through a pad of
Celite.RTM. and concentrated under reduced pressure to afford
(trans)-4-(4-amino-H-pyrazol-1-yl)cyclohexanol (0.13 g, 91% yield)
as a white solid. LC/MS (Table 1, Method w) R.sub.t=0.162 min.; MS
m/z: 182 (M+H).sup.+.
Step D: (cis)-4-(4-amino-1H-pyrazol-1-yl)cyclohexanol
##STR00047##
[0331] To a solution of
(cis)-4-(4-nitro-1H-pyrazol-1-yl)cyclohexanol (0.15 g, 0.71 mmol)
in THF (15 mL) was added Raney nickel (0.3 g). The reaction mixture
stirred at about 20.degree. C. for about 3 h, under an atmosphere
of hydrogen. The reaction mixture was filtered through a pad of
Celite.RTM. and concentrated under reduced pressure to afford
(cis)-4-(4-amino-1H-pyrazol-1-yl)cyclohexanol (0.13 g, 91% yield)
as a white solid. LC/MS (Table 1, Method w) R.sub.t=0.210 min.; MS
m/z: 182 (M+H).sup.+.
Preparation #7: (trans)-3-(4-amino-1H-pyrazol-1-yl)cyclohexanol
##STR00048##
[0332] Step A: 3-((tert-butyldimethylsilyl)oxy)cyclohexanol
##STR00049##
[0334] To a solution cyclohexane-1,3-diol (10 g, 86 mmol) and
imidazole (8.9 g, 130 mmol) in THF (300 mL) was added
tert-butylchlorodimethylsilane (5.2 g, 34.4 mmol). The reaction
mixture stirred at about 15.degree. C. for about 12 h. The solvent
was removed under reduced pressure and the residue was diluted with
water (100 mL). The material was extracted with EtOAc (2.times.200
mL), dried over anhydrous Na.sub.2SO.sub.4, filtered, and
concentrated under reduced pressure to afford
3-((tert-butyldimethylsilyl)oxy)cyclohexanol (7.8 g, 37% yield) as
a light yellow oil. .sup.1H NMR (400 MHz, CHLOROFORM-d)
.delta.=4.12-3.98 (m, 1H), 3.95-3.77 (m, 1H), 1.95-1.74 (m, 2H),
1.72-1.64 (m, 1H), 1.62-1.49 (m, 3H), 1.48-1.40 (m, 1H), 1.35-1.26
(m, 1H), 0.89-0.86 (m, 9H), 0.06 (d, J=3.1 Hz, 3H), 0.04--0.01 (m,
3H)
Step B:
(trans)-3-((tert-butyldimethylsilyl)oxy)cyclohexyl)-4-nitro-1H-pyr-
azole and
(cis)-3-((tert-butyldimethylsilyl)oxy)cyclohexyl)-4-nitro-1H-pyr-
azole
##STR00050##
[0336] To a solution of 4-nitro-1H-pyrazole (3.6 g, 32.2 mmol),
3-((tert-butyldimethylsilyl)oxy)cyclohexanol (7.8 g, 32.2 mmol) and
triphenylphosphine (12.6 g, 48.2 mmol) in THF (250 mL) was added a
solution of DIAD (9.4 mL, 48.2 mmol) in THF (50 mL) at about
0.degree. C. The reaction mixture warmed to about 15.degree. C. and
stirred for about 12 h. The solvent was removed under reduced
pressure and the remaining residue was purified via silicagel
chromatography eluting with petroleum ether/EtOAc (50:1 to 30:1) to
give
(trans)-3-((tert-butyldimethylsilyl)oxy)cyclohexyl)-4-nitro-1H-pyrazole
(3.1 g, 29% yield). .sup.1H NMR (400 MHz, CHLOROFORM-d)
.delta.=8.14 (s, 1H), 8.07 (s, 1H), 4.56 (tt, J=3.7, 11.7 Hz, 1H),
4.28 (br. s., 1H), 2.13 (dd, J=1.5, 10.8 Hz, 2H), 1.96-1.86 (m,
2H), 1.78-1.66 (m, 3H), 1.50-1.40 (m, 1H), 0.92 (s, 9H), 0.10-0.04
(m, 6H) and
(cis)-3-((tert-butyldimethylsilyl)oxy)cyclohexyl)-4-nitro-1H-pyrazole
(1.84 g, 17% yield) .sup.1H NMR (400 MHz, CHLOROFORM-d)
.delta.=8.17 (s, 1H), 8.07 (s, 1H), 4.17 (tt, J=3.9, 12.0 Hz, 1H),
3.77-3.68 (m, 1H), 2.40-2.32 (m, 1H), 2.15 (d, J=12.3 Hz, 1H), 1.94
(dt, J=3.5, 6.6 Hz, 2H), 1.80-1.70 (m, 1H), 1.60 (dq, J=3.1, 12.2
Hz, 1H), 1.47-1.28 (m, 2H), 0.88 (s, 9H), 0.13--0.01 (m, 6H).
Step C: (trans)-3-(4-nitro-1H-pyrazol-1-yl)cyclohexanol
##STR00051##
[0338] To a solution of of
(trans)-3-((tert-butyldimethylsilyl)oxy)cyclohexyl)-4-nitro-1H-pyrazole
(1 g, 3.07 mmol) in MeOH (30 mL) was added a solution of HI (0.513
ml, 3.07 mmol) in MeOH (5 mL). The reaction mixture stirred at
about 15.degree. C. for about 12 h. The solvent was removed under
reduced pressure and the remaining residue was partitioned between
saturated aq. NaHCO.sub.3 (30 mL) and EtOAc (100 mL). The organic
layer was washed with saturated aq. sodium thiosulfate (2.times.30
mL), dried over Na.sub.2SO.sub.4, filtered, and concentrated under
reduced pressure to give
(trans)-3-(4-nitro-1H-pyrazol-1-yl)cyclohexanol (0.64 g, 94% yield)
as a yellow solid.
[0339] LC/MS (Table 1, Method w) R.sub.t=1.07 min.; MS m/z: 212
(M+H).sup.+.
Step D: (trans)-3-(4-amino-1H-pyrazol-1-yl)cyclohexanol
##STR00052##
[0341] To a solution of of
(trans)-3-(4-nitro-1H-pyrazol-1-yl)cyclohexanol (0.3 g, 1.42 mmol)
in THF (50 mL) was added Raney nickel (1 g, 17.04 mmol). The
reaction mixture stirred at about 15.degree. C. for about 12 h,
under an atmosphere of hydrogen. The reaction mixture was filtered
through a pad of Celite.RTM. and concentrated under reduced
pressure to afford (trans)-3-(4-amino-1H-pyrazol-1-yl)cyclohexanol
(0.23 g, 85% yield) as a light red solid. LC/MS (Table 1, Method w)
R.sub.t=0.198 min.; MS m/z: 182 (M+H).sup.+.
Preparation #8: (cis)-3-(4-amino-1H-pyrazol-1-yl)cyclohexanol
##STR00053##
[0342] Step A: (cis)-3-(4-nitro-1H-pyrazol-1-yl)cyclohexanol
##STR00054##
[0344] To a solution of
(cis)-3-((tert-butyldimethylsilyl)oxy)cyclohexyl)-4-nitro-1H-pyrazole
(0.9 g, 2.77 mmol, Preparation #7, Step B) in MeOH (30 mL) was
added a solution of HI (0.54 ml, 3.32 mmol) in MeOH (5 mL). The
reaction mixture stirred at about 15.degree. C. for about 12 h. The
solvent was removed under reduced pressure and the remaining
residue was partitioned between saturated aq. NaHCO.sub.3 (30 mL)
and EtOAc (100 mL). The organic layer was washed with saturated aq.
sodium thiosulfate (2.times.30 mL), dried over Na.sub.2SO.sub.4,
filtered, and concentrated under reduced pressure to give
(cis)-3-(4-nitro-1H-pyrazol-1-yl)cyclohexanol (0.54 g, 88% yield).
LC/MS (Table 1, Method w) R.sub.t=1.07 min.; MS n/z: 212
(M+H).sup.+.
Step B: (cis)-3-(4-amino-1H-pyrazol-1-yl)cyclohexanol
##STR00055##
[0346] To a solution of
(trans)-3-(4-nitro-1H-pyrazol-1-yl)cyclohexanol (0.54 g, 2.56 mmol)
in THF (50 mL) was added Raney nickel (1 g, 17.04 mmol). The
reaction mixture stirred at about 15.degree. C. for about 12 h,
under an atmosphere of hydrogen. The reaction mixture was filtered
through a pad of Celite.RTM. and concentrated under reduced
pressure to afford (cis)-3-(4-amino-H-pyrazol-1-yl)cyclohexanol
(0.41 g, 87% yield) as a pink solid. LC/MS (Table 1, Method w)
R.sub.t=0.232 min.; MS m/z: 182 (M+H).sup.+.
Preparation #9: ethyl
4-(4-amino-1H-pyrazol-1-yl)cyclohexanecarboxylate
##STR00056##
[0347] Step A: ethyl
4-(4-nitro-1H-pyrazol-1-yl)cyclohexanecarboxylate
##STR00057##
[0349] A round bottom flask was charged with ethyl
4-hydroxycyclohexanecarboxylate (4.1 g, 23.8 mmol),
4-nitro-1H-pyrazole (2.2 g, 19.0 mmol), and PPh.sub.3 (6.2 g, 23.8
mmol) in THF (95 mL). The reaction was degassed with nitrogen for
about 5 min. before the addition of DIAD (7.4 ml, 38.1 mmol). The
reaction mixture was stirred at about 60.degree. C. for about 16 h.
The solvent was concentrated under reduced pressure. The crude
material was purified via silicagel chromatography eluting with
EtOAc/Heptanes (0-100%) to afford ethyl
4-(4-nitro-H-pyrazol-1-yl)cyclohexanecarboxylate (5.0 g, quant
yield). LC/MS (Table 1, Method h) R.sub.t=2.22 min.; MS m/z: 268
(M+H).sup.+.
Step B: ethyl 4-(4-amino-1H-pyrazol-1-yl)cyclohexanecarboxylate
##STR00058##
[0351] A solution of ethyl
4-(4-nitro-1H-pyrazol-1-yl)cyclohexanecarboxylate (5.1 g, 18.9
mmol) in EtOH (95 mL) was passed through the H-Cube with a 10% Pd/C
cartridge under 10 bar of hydrogen at 50.degree. C. The reaction
solution cycled through the reactor for 5 h. The solvent was
concentrated under reduced pressure to afford ethyl
4-(4-amino-1H-pyrazol-1-yl)cyclohexanecarboxylate (4.5 g, quant
yield). LC/MS (Table 1, Method h) R.sub.t=1.30 min.; MS m/z: 238
(M+H).sup.+.
Preparation #10: tert-butyl
4-(4-amino-1H-pyrazol-1-yl)-2,6-dimethylpiperidine-1-carboxylate
##STR00059##
[0352] Step A: dimethyl
2,6-dimethyl-4-oxopiperidine-3,5-dicarboxylate
##STR00060##
[0354] To a mixture of dimethyl 3-oxopentanedioate (100 g, 574
mmol) and acetaldehyde (65.8 g, 1.5 mol) was bubbled ammonia gas at
-30.degree. C. until the liquid was saturated. The solution was
stored in the freezer for about 20 h. The yellow reside was
purified by chromatography on silica gel eluting with (PE:
EtOAc=10:1 to PE:EtOAc=1:2) to give dimethyl
2,6-dimethyl-4-oxopiperidine-3,5-dicarboxylate (120 g, 50% yield,
60% purity). .sup.1H NMR (400 MHz, CHLOROFORM-d) 3.73-3.70 (m, 9H),
3.39-3.31 (m, 2H), 3.04-2.98 (m, 2H), 1.19 (d, J=6.2 Hz, 6H).
Step B: 2,6-dimethylpiperidin-4-one hydrochloride
##STR00061##
[0356] A solution of dimethyl
2,6-dimethyl-4-oxopiperidine-3,5-dicarboxylate (120 g, 286 mmol) in
10% aq. HCl (900 mL) was heated to about 110.degree. C. for 24 hrs.
The solvent was concentrated under reduced pressure to give
2,6-dimethylpiperidin-4-one hydrochloride (50 g, 80% yield), which
was used for the next step directly. .sup.1H NMR (400 MHz,
DMSO-d.sub.6) 10.15-9.80 (m, 2H), 3.82 (d, J=4.4 Hz, 1H), 3.59-3.45
(m, 1H), 2.75-2.60 (m, 2H), 2.47-2.34 (m, 2H), 1.38-1.27 (m,
6H)
Step C: tert-butyl 2,6-dimethyl-4-oxopiperidine-1-carboxylate
##STR00062##
[0358] To a solution of 2,6-dimethylpiperidin-4-one hydrochloride
(50 g, 281 mmol) in 1,4-dioxane (350 mL) and water (350 mL) was
added Na.sub.2CO.sub.3 (59 g, 562 mmol) portionwise. Boc-anhydride
(123 g, 562 mmol) was added and the resulting reaction mixture was
stirred at about 15.degree. C. for about 24 h. The solvent was
concentrated under reduced pressure. The resulting residue was
extracted with MTBE (3.times.400 mL) and the organic layer was
washed with brine (300 mL) and dried over anhydrous
Na.sub.2SO.sub.4. After evaporation, the crude product was purified
by column chromatography on silica gel eluting with (PE to
PE:EA=10:1) to afford tert-butyl
2,6-dimethyl-4-oxopiperidine-1-carboxylate (25 g, 38% yield), which
was a mixture of Trans/Cis=2:1.
[0359] .sup.1H NMR (400 MHz, CHLOROFORM-d) 4.66 (dd, J=4.9, 6.8 Hz,
2H), 4.32 (t, J=6.3 Hz, 1H), 2.79 (dd, J=6.5, 17.8 Hz, 1H), 2.66
(dd, J=7.6, 14.7 Hz, 2H), 2.31 (dd, J=1.6, 17.6 Hz, 1H), 2.24-2.17
(m, 2H), 1.43 (d, J=2.3 Hz, 15H), 1.23-1.17 (m, 10H).
Step D: tert-butyl
4-hydroxy-2,6-dimethylpiperidine-1-carboxylate
##STR00063##
[0361] To a solution of tert-butyl
2,6-dimethyl-4-oxopiperidine-1-carboxylate (25 g, 110 mmol) in EtOH
(75 mL) was added NaBH.sub.4 (6.24 g, 165 mmol) at about 0.degree.
C. The resulting solution was warmed to about 15.degree. C. and
stirred for about 4 h. The reaction mixture was cooled to about
0.degree. C. before the addition of saturated aq. NH.sub.4Cl (80
mL). The solvent was evaporated under reduced pressure and EtOAc
(100 mL) was added to the residue. The two layers were separated
and the aqueous layer was extracted with EtOAc (3.times.100 mL).
The combined organic layers were dried over anhydrous
Na.sub.2SO.sub.4, and evaporated to give tert-butyl
4-hydroxy-2,6-dimethylpiperidine-1-carboxylate (20 g, 79% yield) as
a colorless oil. .sup.1H NMR (400 MHz, CHLOROFORM-d) 4.47-4.37 (m,
1H), 4.31-4.13 (m, 2H), 4.01 (br. s., 1H), 3.94-3.82 (m, 1H),
2.25-2.14 (m, 1H), 2.09-1.99 (m, 2H), 1.98-1.79 (m, 2H), 1.63-1.53
(m, 2H), 1.45 (s, 13H), 1.39-1.28 (m, 6H), 1.23-1.12 (m, 2H).
Step E: tert-butyl
2,6-dimethyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylate
##STR00064##
[0363] To a solution of tert-butyl
4-hydroxy-2,6-dimethylpiperidine-1-carboxylate (8.9 g, 38.8 mmol),
4-nitro-1H-pyrazole (4.39 g, 38.8 mmol), and PPh.sub.3 (15.3 g,
58.2 mmol) in THF (150 ml) was added DIAD (11.3 mL, 58.2 mmol)
dropwise. The reaction mixture stirred at about 0.degree. C. for 10
minutes then warmed to about 30.degree. C. and stirred for about 16
hrs. The reaction mixture was partitioned between EtOAc and brine.
The organic portion was dried over anhydrous Na.sub.2SO.sub.4,
filtered, and concentrated under reduced pressure. The crude
material was purified via silicagel chromatography eluting with
EtOAc/Petroleum ether (0-10%) to afford tert-butyl
2,6-dimethyl-4-(4-nitro-H-pyrazol-1-yl)piperidine-1-carboxylate (7
g, 56% yield) as a white solid.
[0364] .sup.1H NMR (400 MHz, CHLOROFORM-d) .delta.=8.19 (s, 1H),
8.10 (d, J=1.6 Hz, 1H), 4.77-4.46 (m, 3H), 3.89-3.76 (m, 1H),
2.43-2.24 (m, 1H), 2.16-1.90 (m, 4H), 1.49 (d, J=1.6 Hz, 11H), 1.46
(d, J=6.7 Hz, 2H), 1.38-1.27 (m, 6H)
Step F: tert-butyl
4-(4-amino-1H-pyrazol-1-yl)-2,6-dimethylpiperidine-1-carboxylate
##STR00065##
[0366] To a solution of tert-butyl
2,6-dimethyl-4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylate (2
g, 6.17 mmol) in THF (50 mL) and NH.sub.4OH (0.5 mL) was added
Raney nickel (1.1 g, 18.1 mmol). The reaction mixture stirred at rt
for about 2 h, under an atmosphere of hydrogen. The reaction
mixture was filtered through a pad of Celite.RTM. and concentrated
under reduced pressure to afford tert-butyl
4-(4-amino-1H-pyrazol-1-yl)-2,6-dimethylpiperidine-1-carboxylate
(1.8 g, 99% yield). LC/MS (Table 1, Method w) R.sub.t=0.96 min.; MS
m/z: 295 (M+H).sup.+.
General Procedure A: Nucleophilic Displacement of an Aryl or
Heteroaryl Halide with an Amine
[0367] To a microwave vessel, a vial, or a round bottom flask is
added an aryl or heteroaryl halide (preferably 1 equiv), an amine
or an amine salt (1-10 equiv, preferably 1.1-2 equiv), a solvent
(such as 1,4-dioxane, MeCN, i-PrOH, n-PrOH, n-BuOH, toluene, DMSO,
DMF, DMA or EtOH, preferably 1,4-dioxane [microwave] or i-PrOH
[thermal heating]), and a base (such as K.sub.2CO.sub.3,
Na.sub.2CO.sub.3, TEA or DIEA, preferably TEA, DIEA, or
K.sub.2CO.sub.3, 1-5 equiv, preferably 2-4 equiv). Optionally the
aryl or heteroaryl halide and the amine or amine salt are each
separately dissolved in a solvent prior to combining. The reaction
mixture is heated at about 40-220.degree. C. thermally (preferably
about 80-100.degree. C.) for about 0.5-36 h (preferably about 8-24
h) or is subjected to microwave heating at about 100-200.degree. C.
(preferably about 130-150.degree. C.) for about 0.5-8 h (preferably
about 0.5-2 h). In cases where the reaction does not proceed to
completion as monitored by TLC, LC/MS, or HPLC, the reaction may be
resubjected to thermal heating at about 40-220.degree. C.
(preferably about 80-100.degree. C.) for about 0.5-8 h (preferably
about 1-2 h) or microwave heating at about 120-200.degree. C.
(preferably about 130-150.degree. C.) for an additional about 1-8 h
(preferably about 0.5-2 h) with the optional addition of more amine
or amine salt (1-10 equiv, preferably 0.5-1.5 equiv) and/or base
(such as K.sub.2CO.sub.3, Na.sub.2CO.sub.3, TEA or DIEA, preferably
TEA, DIEA or K.sub.2CO.sub.3, 1-5 equiv, preferably 2-4 equiv).
This process is repeated until the reaction proceeds no further.
After cooling to ambient temperature, the reaction is worked up
using one of the following methods. Method 1: The reaction is
concentrated under reduced pressure. Method 2: A reaction mixture
containing a precipitate may be filtered to collect the target
compound, while optionally washing with organic solvent or solvents
such as Et.sub.2O, DCM and/or petroleum ether. Method 3: The
reaction mixture is diluted with an organic solvent such as MeOH,
silica gel is added, and the mixture is concentrated under reduced
pressure to prepare for separation by chromatography with solid
loading. Method 4: The reaction mixture is concentrated under
reduced pressure prior to the addition of an organic solvent such
as EtOAc or DCM and is then optionally washed with water and/or
brine, dried over anhydrous Na.sub.2SO.sub.4 or MgSO.sub.4,
filtered or decanted, and concentrated under reduced pressure.
Method 5: An organic solvent such as EtOAc or DCM is added with the
optional addition of water or brine and the layers are separated.
The aqueous layer is then optionally extracted with additional
organic solvent such as EtOAc or DCM. The combined organic layers
are optionally washed with brine or water, dried over anhydrous
MgSO.sub.4 or Na.sub.2SO.sub.4, filtered or decanted, and
concentrated under reduced pressure.
Illustration of General Procedure A
Preparation #A.1:
8-((6-bromo-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-dihydro-1H-benz-
o [b]azepin-2(3H)-one
##STR00066##
[0369] To a solution of 6,8-dibromo-[1,2,4]triazolo[1,5-a]pyrazine
(0.200 g, 0.720 mmol, Ark Pharm) and
8-amino-4,5-dihydro-1H-benzo[b]-azepin-2(3H)-one (0.139 g, 0.792
mmol, Astatech) in i-PrOH (8 mL) was added DIEA (0.377 mL, 2.16
mmol). The mixture was heated to reflux for about 24 h. The
resulting solution was cooled to rt and filtered to give
8-((6-bromo-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-dihydro-1H-benz-
o[b]azepin-2(3H)-one (0.15 g, 56%) as a brown solid: .sup.1H NMR
(DMSO-d6) .delta. 9.62 (s, 1H), 8.69 (s, 1H), 8.59 (s, 1H),
7.61-7.67 (m, 2H), 7.23 (d, J=8.4 Hz, 1H), 2.65 (t, J=6.4 Hz, 2H),
2.07-2.15 (m, 3H), 1.23 (m, 1H).
TABLE-US-00006 TABLE A.1 Examples prepared from
8-bromo-6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazine [Example #11,
Step E] as described in General Procedure A. R.sub.t min m/z CSF-1R
(Table 1, ESI+ Enzyme Amine Product Example Method) (M + H).sup.+
IC.sub.50 4-(4-amino-1H-pyrazol-1-yl)- 2-methylbutan-2-ol
[Preparation # 4] ##STR00067## A.1.1 3.15 (w) 370 A
1-(tetrahydro-2H-pyran-4-yl)- 1H-pyrazol-4-aminc [WO2011/71716A1]
##STR00068## A.1.2 3.26 (w) 368 A 1-(2,2,6,6-
tetramethyltetrahydro-2H- pyran-4-yl)-1H-pyrazol-4- amine
[Preparation #5] ##STR00069## A.1.3 3.42 (v) 356 A
1-(oxetan-3-yl)-1H-pyrazol-4- amine [WO2014/194242A2] ##STR00070##
A.1.4 3.14 (w) 340 A (trans)-4-(4-amino-1H-
pyrazol-1-yl)cyclohexanol [Preparation #6] ##STR00071## A.1.5 3.07
(v) 356 A (cis)-4-(4-amino-1H-pyrazol- 1-yl)cyclohexanol
[Preparation #6] ##STR00072## A.1.6 3.14 (v) 356 A
1H-pyrazol-4-amine [CombiBlocks] ##STR00073## A.1.7 1.96 (h) 284 A
ethyl 4-(4-amino-1H-pyrazol- 1-yl)cyclohexanecarboxylate
[Preparation #9] ##STR00074## A.1.8 2.90 (h) 438 A
General Procedure B: Buchwald-Hartwig Reaction of an Aryl or
Heteroaryl Halide with an Amine
[0370] A mixture of an aryl or heteroaryl halide (1.0 equiv), an
amine (1 to 2.2 equiv, preferably 1 to 1.2 equiv), a palladium
catalyst (such as Pd.sub.2dba.sub.3, Pd(OAc).sub.2, preferably
Pd(OAc).sub.2; 0.01 to 1.0 equiv, preferably 0.04 to 0.1 equiv), a
ligand (such as Xphos, Xantphos or tert-butyl-X-phos, preferably
Xantphos or XPhos, 0.01 to 2.0 equiv, preferably 0.04 to 0.1 equiv)
and a base (such as K.sub.2CO.sub.3, Na.sub.2CO.sub.3,
Cs.sub.2CO.sub.3, K.sub.3PO.sub.4, NaOt-Bu, KOt-Bu, KOAc, KOH,
preferably Cs.sub.2CO.sub.3 or K.sub.2CO.sub.3; 1 to 5 equiv,
preferably 1 to 3 equiv) are added to a solvent (such as
1,4-dioxane, t-BuOH, preferably t-BuOH). The mixture is degassed
under an inert atmosphere (such as nitrogen or argon, preferably
nitrogen) and heated with conventional or microwave heating at
about 80 to 150.degree. C. (preferably about 85 to 95.degree. C.)
for about 2 to 24 h (preferably about 16 h). The mixture is cooled
to rt. The mixture is optionally filtered through a media (such as
silica gel or Celite.RTM.) which is rinsed with an appropriate
solvent (such as EtOAc, 1,4-dioxane, THF, MeCN, DCM, Et.sub.2O,
MeOH, EtOH, DMSO, 1:1 MeOH/DMSO or 2:1 MeOH/DMSO, preferably
MeOH/DMSO) and then the filtrate is optionally concentrated under
reduced pressure or under a warm nitrogen stream to give a
residue.
Illustration of General Procedure B
Preparation #B.1:
6-chloro-N-(1-methyl-1H-pyrazol-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-ami-
ne
##STR00075##
[0372] To a microwave vial was added
8-bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine (0.70 g, 3.0 mmol,
Example #1, Step A), 1-methyl-1H-pyrazol-3-amine (0.32 g, 3.3 mmol,
Matrix Scientific), Xantphos (0.37 g, 0.63 mmol), Pd(OAc).sub.2
(0.068 g, 0.30 mmol), cesium carbonate (2.453 g, 7.53 mmol) and
1,4-dioxane (7 mL). The mixture was then heated in a microwave for
about 1.5 h at about 120.degree. C. The reaction mixture was cooled
to rt, filtered, washed with DCM and MeOH. Silica gel (2 g) was
added to the filtrate. The mixture was concentrated under reduced
pressure and purified via silica gel chromatography eluting with
0-50% DCM/MeOH/NH.sub.4OH (90:9:1) in DCM. The product-containing
fractions were combined and concentrated under reduced pressure and
dried under vacuum at about 55.degree. C. to give
6-chloro-N-(1--methyl-1H-pyrazol-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-
-8-amine (0.55 g, 73%, 75% purity by NMR): LC/MS (Table 1, Method
a) R.sub.t=1.29 min.; MS m/z: 249 (M+H).sup.+.
General Procedure C: Reaction of an Aryl or Heteroaryl Halide with
a Boronic Acid or Boronate Ester
[0373] To a mixture of an aryl halide (preferably 1 equiv), a
boronic acid or boronate ester (1-2.5 equiv, preferably 1.3-2.0
equiv), and an inorganic base (for example, potassium fluoride,
sodium carbonate or cesium carbonate, preferably cesium carbonate;
1.1-16 equiv, preferably 2-3 equiv) in a solvent (for example THF,
DME, DMF, 1,4-dioxane, DME/water, 1,4-dioxane/water,
toluene/EtOH/water, THF/MeOH/water or 1,4-dioxane/EtOH/water;
preferably THF/MeOH/water, 1,4-dioxane/EtOH/water or DME) is added
a palladium catalyst (for example
tris(benzylideneacetone)dipalladium(0),
tetrakis(triphenylphosphine)palladium(0),
bis(acetato)-triphenylphosphinepalladium(II), polymer-bound
FibreCat.TM. 1032, SiliaCat DPP-Pd [Silicycle],
(1,1'-bis(diphenyl-phosphino)ferrocene)dichloropalladium(II), or
dichlorobis(triphenyl-phosphine)palladium(II); preferably
tris(bcnzylideneacetone)di-palladium(0) or
tetrakis-(triphenylphosphine)palladium(0), 0.01-0.20 equiv,
preferably 0.1 equiv) and optionally a ligand (for example
tricyclohexylphosphine, XPhos, Xantphos, tert-butyl-XPhos, or
tri-t-butyl-phosphane; preferably no ligand, XPhos, or Xantphos
(0.01-1.0 equiv, preferably 0.1-0.2 equiv) is added. The reaction
mixture is heated at about 40-120.degree. C. (preferably about
80-90.degree. C.) for about 1-24 h (preferably about 2 h)
thermally, or at about 100-200.degree. C. (preferably about
120-150.degree. C.) for about 5 min-3 h (preferably about 30 min)
in a microwave. In cases where the reaction does not proceed to
completion as monitored by TLC, HPLC, or LCMS, additional reagents
and reactants can be optionally added and the reaction can be
resubjected to heating for an additional about 5 min-3 h
(preferably about 30 min) in the microwave or 1-24 h (preferably
about 2 h) thermally at the same or higher temperature via the same
or different heating method. This process is repeated until the
reaction proceeds no further. The reaction mixture is allowed to
cool to ambient temperature and is worked up using one of the
following methods. Method 1. For reactions containing water, the
reaction mixture may be optionally filtered then diluted with an
organic solvent (such as DCM or EtOAc). The layers are separated,
the organic solution is optionally washed with water and/or brine,
dried over MgSO.sub.4 or Na.sub.2SO.sub.4, filtered, and the
solvent is removed under reduced pressure. Method 2. The reaction
mixture is concentrated under reduced pressure. Method 3. The
catalyst is removed by filtration and the filtrate is concentrated
under reduced pressure or the filtrate is washed with water and/or
brine, the layers are separated, the organic solution is dried over
MgSO.sub.4 or Na.sub.2SO.sub.4, filtered, and the solvent is
removed under reduced pressure. Method 4. Water and/or MeOH is
added and the resulting precipitate is collected via
filtration.
Illustration of General Procedure C
Preparation #C.1: tert-butyl
3-(8-((2-oxo-2,3,4,5-tetrahydro-1H-benzo[b]azepin-8-yl)amino)-[1,2,4]tria-
zolo[1,5-a]pyrazin-6-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate
##STR00076##
[0375] To a solution of
8-((6-bromo-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-dihydro-1H-benz-
o [b]azepin-2(3H)-one (0.500 g, 1.34 mmol, Preparation #A.1) in DME
(12 mL) was added tert-butyl
3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro-1H-pyrrole-1--
carboxylate (0.791 g, 2.68 mmol, Combi-Blocks), XPhos (0.058 mg,
0.134 mmol), tris(benzylideneacetone)di-palladium(0) (0.123 g,
0.134 mmol) and Cs.sub.2CO.sub.3 (1.30 g, 4.02 mmol). The mixture
was heated at about 140.degree. C. for about 30 min in a microwave
reactor. The resulting solution was filtered and the solid was
washed with DCM then washed with water and brine. The organic phase
was dried over Na.sub.2SO.sub.4 and concentrated to give a crude
solid, which was washed with MeOH to afford tert-butyl
3-(8-((2-oxo-2,3,4,5-tetrahydro-1H-benzo
[b]azepin-8-yl)amino)-[1,2,4]triazolo[,
5-a]pyrazin-6-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (0.25 g,
40%) as a brown solid: .sup.1H NMR (DMSO-d6) .delta. 10.05 (s, 1H),
9.65 (d, J=7.9 Hz, 1H), 8.60 (s, 1H), 8.50-8.38 (m, 1H), 7.82 (d,
J=12.3 Hz, 1H), 7.71 (t, J=9.5 Hz, 1H), 7.20 (t, J=6.6 Hz, 1H),
6.68 (d, J=11.9 Hz, 1H), 4.44 (br s, 2H), 4.24 (br s, 2H), 2.64 (br
s, 2H), 2.24-1.96 (m, 4H), 1.59-1.35 (s, 9H).
TABLE-US-00007 TABLE C.1 Examples prepared from
6-bromo-N-(4-morpholinophenyl)-[1,2,4] triazolo[1,5-
a]pyrazin-8-amine (prepared using B from
6,8-dibromo-[1,2,4]triazolo[1,5-a]pyrazine [Ark Pharm] and
4-morpholinoaniline) using General Procedure C R.sub.t min m/z Btk
Example (Table 1, ESI + Enzyme Boronic acid or boronate Product #
Method) (M + H) IC.sub.50 N-(3-(4,4,5,5-tetramethyl-
1,3,2-dioxaborolan-2- yl)benzyl)acrylamide (prepared using F from
(3- (4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-
yl)phenyl)methanamine [ChemMaker]and acryloyl chloride)
##STR00077## C.1.1 1.59 (b) 546 C
General Procedure D: Acidic Cleavage of a Boc-Protected Amine
[0376] To a solution of an N-Boc amine (1 equiv) in an organic
solvent (such as DCM, DCE, EtOAc, 1,4-dioxane or MeOH, preferably
DCM, EtOAc, MeOH or 1,4-dioxane) is added an acid (such as TFA or
HCl (HCl could be commercially purchased or generated in situ with
MeOH and acetyl chloride), preferably HCl; 2 to 100 equiv,
preferably 25 to 50 equiv). The mixture is stirred at about 0 to
100.degree. C. (preferably about 20 to 60.degree. C.) for about 1
to 24 h (preferably about 1 to 12 h). Optionally, additional acid
(2 to 35 equiv, preferably 20 to 25 equiv) may be added and the
mixture stirred at about 0 to 100.degree. C. (preferably about 20
to 60.degree. C.) for about 1 to 24 h (preferably about 1 to 6 h).
If a solid is present in the mixture, the mixture may be optionally
filtered and the solid washed with an organic solvent such as
1,4-dioxane or Et.sub.2O. The resulting solid is then optionally
dried under reduced pressure to give the targeted compound.
Alternatively, the mixture may be optionally concentrated in vacuo
to give final compound. Alternatively, either the residue or the
solution may be optionally partitioned between water and an organic
solvent (such as EtOAc, Et.sub.2O or DCM). The organic layer is
isolated and may be optionally washed in no particular order with
water and/or aqueous solutions containing a base (such as
NaHCO.sub.3, Na.sub.2CO.sub.3, NaOH, KOH or NH.sub.4OH) and/or
aqueous solutions containing an inorganic salt (such as NaC,
Na.sub.2SO.sub.3 or Na.sub.2S.sub.2O.sub.3). The organic solution
may then be optionally dried with a drying agent (such as anhydrous
MgSO.sub.4 or Na.sub.2SO.sub.4), filtered and concentrated in vacuo
to give the targeted compound.
Illustration of General Procedure D
Preparation #D.1:
8-((6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-dih-
ydro-1H-benzo[b]azepin-2(3H)-one
##STR00078##
[0378] To a solution of
3-[8-(2-oxo-2,3,4,5-tetrahydro-1H-benzo[b]azepin-8-ylamino)-[1,2,4]triazo-
lo[1,5-a]pyrazin-6-yl]-pyrrolidine-1-carboxylic acid methylamide
(1.60 g, 3.45 mmol, Preparation #E.1) in EtOAc (20 mL) was added
HCl/EtOAc (20 mL). The mixture was stirred for about 12 h at about
25.degree. C. The resulting solution was concentrated under reduced
pressure to give
8-((6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-dih-
ydro-1H-benzo[b]azepin-2(3H)-one (1.20 g, 96%) as a brown solid:
LC/MS (Table 1, Method d) R.sub.t=2.12 min; MS m/z: 364
(M+H).sup.+.
TABLE-US-00008 TABLE D.1 Examples prepared using HCl as described
in General Procedure D R.sub.t min m/z CSF-1R Example (Table 1,
ESI+ Enzyme Carbamate Product # Method) (M + H).sup.+ IC.sub.50
tert-butyl 4-(4-((6- cyclohexyl- [1,2,4]triazolo[1,5-a]pyrazin-
8-yl)amino)-1H-pyrazol-1- yl)piperidine-1-carboxylate (prepared
using A from 6,8-dibromo- [1,2,4-]triazolo [1,5 -a]pyrazine
[ArkPharm] and tert-butyl 4-(4-amino-1H- pyrazol-1-yl)piperidine-1-
carboxylate [Combiblocks], C with 2-(cyclohex-1-en-1-yl)-
4,4,5,5-tetramethyl-1,3,2- dioxaborolane [Combiblocks], and E with
Pd(OH).sub.2/C. ##STR00079## D.1.1 1.53(h) 367 A tert-butyl
4-(4-((6-(cyclopent- 1-en-1-yl)-[1,2,4]triazolo[1,5-
a]pyrazin-8-yl)amino)-1H- pyrazol-1-yl)piperidinc-1- carboxylate
(prepared using A from 6,8-dibromo- [1,2,4]triazolo[1,5-a]pyrazine
[ArkPharm] and tert-butyl 4-(4-amino-1H- pyrazol-1-yl)piperidine-1-
carboxylate [Combiblocks], C with 2-(cyclopent-l-en-l-yl)-
4,4,5,5-tetramethyl-1,3,2- dioxaborolane [ArkPharm], and E with
Pd(OH).sub.2/C. ##STR00080## D.1.2 1.42(h) 353 A
General Procedure E: Hydrogenation of a Double Bond
[0379] A reaction vessel is charged with an alkene (1 equiv), neat
or as a solution in an organic solvent or mixture of solvents (such
as THF, EtOAc, MeOH, EtOH or MeOH/AcOH, preferably THF or MeOH)
followed by addition of Pd(OH).sub.2 or Pd/C (0.005-3 equiv,
preferably 2 equiv) either as a solid or in an organic solvent or
mixture of solvents (such as AcOH, THF, EtOAc, MeOH, EtOH or
MeOH/AcOH, preferably AcOH. Optionally the alkene is added to the
Pd mixture. The reaction mixture is sparged with hydrogen. The
mixture is stirred or shaken (preferably stirred when atmospheric
hydrogen is used or shaken when higher pressures of hydrogen is
used) under hydrogen at about atmospheric pressure to 60 psi
(preferably about 50 psi) at about 20-60.degree. C. (preferably
ambient temperature) for about 0.5-5 days (preferably about 1-3
days). The reaction mixture is filtered through a pad of
Celite.RTM. or a nylon membrane. The filter cake is rinsed with an
organic solvent (such as THF, EtOAc, DCM, MeOH, or EtOH, preferably
the reaction solvent) and the filtrate is concentrated under
reduced pressure to give the targeted compound.
[0380] The reaction could also be carried in flow chemistry style,
using H-cube, with same choices of solutions listed above. The
reaction mixture as a solution is passed through Catcart.RTM.
filled with Pd(OH).sub.2/C or Pd/C, under hydrogen from 1 bar to 80
bar (preferably about 40-60 bar) at about 20-80.degree. C.
(preferably 30-60.degree. C.) for about 0.5-24 h (preferably about
8 h). The mixture is then concentrated under reduced pressure to
give the targeted compound.
Illustration of General Procedure E
Preparation #E.1: tert-butyl
3-(8-((2-oxo-2,3,4,5-tetrahydro-1H-benzo[b]azepin-8-yl)amino)-[1,2,4]tria-
zolo[1,5-a]pyrazin-6-yl)pyrrolidine-1-carboxylate
##STR00081##
[0382] To solution of
3-[8-(2-oxo-2,3,4,5-tetrahydro-1H-benzo[b]azepin-8-ylamino)-[1,2,4]triazo-
lo[1,5-a]pyrazin-6-yl]-2,5-dihydro-pyrrole-1-carboxylic acid
tert-butyl ester (2.0 g, 4.33 mmol, Preparation #C.1) in MeOH (300
mL) and THF (60 mL) was added 10% Pd/C (2 g) and AcOH (30 mL). The
suspension was stirred for about 72 h at about 25.degree. C. under
H.sub.2 (50 psi) atmosphere. The resulting solution was filtered
through Celite.RTM. and concentrated under reduced pressure to give
tert-butyl
3-(8-((2-oxo-2,3,4,5-tetrahydro-H-benzo[b]azepin-8-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyrazin-6-yl)pyrrolidine-1-carboxylate (1.8 g, 90%):
LC/MS (Table 1, Method c) R.sub.t=1.40 min; MS m/z: 464
(M+H).sup.+.
TABLE-US-00009 TABLE E.1 Examples prepared using Pd/C or
Pd(OH).sub.2/C as descibed in General Procedure E R.sub.t min m/z
CSF-1R Example (Table 1, ESO + Enzyme Olefin Product # Method) (M +
H).sup.+ IC.sub.50 6-(cyclohex-1-en-1-yl)-N-(1-
isopropyl-1H-pyrazol-4-yl)- [1,2,4]triazolo[1,5-a]pyrazin-8- amine
(prepared using A from 6,8-dibromo- [1,2,4]triazolo[1,5-a]pyrazinc
[ArkPharm] and 1-isopropyl-1H-pyrazol-4- amine [Combiblocks], C
with 2-(cyclohex-1-en-1-yl)-4,4,5,5- tetramethyl-1,3,2-
dioxaborolane [Combiblocks] ##STR00082## E.1.1 2.55 (h) 326 A
6-(4,4-dimethylcyclohex-1-en- 1-yl)-N-(1-methyl-1H-pyrazol-
4-yl)-[1,2,4]triazolo[1,5- a]pyrazin-8-amine (prepared using A from
6,8-dibromo- [1,2,4]triazolo[1,5-a]pyrazine [ArkPharm] and
1-methyl-1H-pyrazol-4-amine, C with 2-(4,4-dimethylcyclohex-1-en-
1-yl)-4,4,5,5-tetramethyl-1,3,2- dioxaborolane ##STR00083## E.1.2
3.22 (t) 326 A N-(1-methyl-1H-pyrazol-4-yl)-
6-(4-methylcyclohex-1-en-1- yl)-[1,2,4]triazolo[1,5-
a]pyrazin-8-amine (Prepared using A from 6,8-dibromo-
[1,2,4]triazolo[1,5-a]pyrazine [ArkPharm] and
1-methyl-1H-pyrazol-4-amine, C with 4,4,5,5-tetramethyl-2-(4-
methylcyclohex-1-en-1-yl)- 1,3,2-dioxaborolane [CombiBlocks]
##STR00084## E.1.3 3.37 (t) 312 A N-(1-methyl-1H-pyrazol-4-yl)-
6-(4-methylcyclohex-1-en-1- yl)-[1,2,4]triazolo[1,5-
a]pyrazin-8-amine (Prepared using A from 6,8-dibromo-
[1,2,4]triazolo[1,5-a]pyrazine [ArkPharm] and
1-methyl-1H-pyrazol-4-amine, C with 4,4,5,5-tetramethyl-2-(4-
methylcyclohex-1-en-1-yl)- 1,3,2-dioxaborolane [CombiBlocks]
##STR00085## E.1.4 3.39 (t) 312 A N-(1-methyl-1H-pyrazol-4-yl)-
6-(4- (trifluoromethyl)cyclohex-1- en-1-yl)-[1,2,4]triazolo[1,5-
a]pyrazin-8-amine (Prepared using A from 6,8-dibromo-
[1,2,4]triazolo[1,5-a]pyrazine [ArkPharm] and
1-methyl-1H-pyrazol-4-amine, C with 4,4,5,5-tetramethyl-2-[4-
(trifluoromethyl)-1- cyclohexen-1-yl]-1,3,2- dioxaborolane
[Acentix] ##STR00086## E.1.5 2.27 (h) 366 A
N-(6-(cyclohex-1-en-1-yl)- [1,2,4]triazolo[1,5-a]pyridin-8-
yl)-6,7-dihydro-4H- pyrazolo[5,1-c][1,4]oxazin-2- amine (Prepared
using B from 8-bromo-6-chloro- [1,2,4]triazolo[1,5-a]pyridine
[Example #1, Step A] and 6,7-dihydro-4H-pyrazolo[5,1-
c][1,4]oxazin-2-amine [Preparation #1], and C with 1-cyclohexenyl
boronic acid [CombiBlocks] ##STR00087## E.1.6 2.23 (h) 339 A
6-(cyclohex-1-en-1-yl)-N-(5- methyl-4,5,6,7-
tetrahydropyrazolo[1,5- a]pyrazin-2-yl)-
[1,2,4]triazolo[1,5-a]pyridin-8- amine (Prepared using B from
8-bromo-6-chloro- [1,2,4]triazolo[1,5-a]pyridine [Example #1, Step
A] and 5-methyl-4,5,6,7- tetrahydropyrazolo[1,5- a]pyrazin-2-amine
[Astatech], and C with 1-cyclohexenyl boronic acid [CombiBlocks]
##STR00088## E.1.7 2.09 (h) 352 A 6-(cyclopent-1-en-1-yl)-N-(1-
methyl-1H-pyrazol-4-yl)- [1,2,4]triazolo[1,5-a]pyrazin-8- amine
(Prepared using A from 6,8-dibromo- [1,2,4]triazolo[1,5-a]pyrazine
[ArkPharm] and 1-methyl-1H-pyrazol-4-amine, C with
2-(cyclopent-1-en-1-yl)- 4,4,5,5-tetramethyl-1,3,2- dioxaborolane
[Combiblocks] ##STR00089## E.1.8 2.82 (x) 284 A
6-(cyclopent-1-en-1-yl)-N(5- methyl-4,5,6,7-
tetrahydropyrazolo[1,5- a]pyrazin-2-yl)-
[1,2,4]triazolo[1,5-a]pyridin-8- amine (Prepared using B from
8-bromo-6-chloro- [1,2,4]triazolo[1,5-a]pyridine [Example #1, Step
A] and 5-methyl-4,5,6,7- tetrahydropyrazolo[1,5- a]pyrazin-2-amine
[Astatech], and C with 2-(cyclopent-1-en-1-yl)-
4,4,5,5-tetramethyl-1,3,2- dioxaborolane [Combiblocks] ##STR00090##
E.1.9 1.80 (h) 338 A 6-(cyclohex-1-en-1-yl)-N-(1-
methyl-1H-pyrazol-4-yl)- [1,2,4]triazolo[1,5-a]pyrazin-8- amine
(Prepared using A from 6,8-dibromo- [1,2,4]triazolo[1,5-a]pyrazine
[ArkPharm] and 1-methyl-1H-pyrazol-4-amine [Astatech], C with
2-(cyclohex-1-en-1-yl)-4,4,5,5- tetramethyl-1,3,2- dioxaborolane
[ArkPharm] ##STR00091## E.1.10 2.22 (h) 298 A
General Procedure F: Formation of an Amide from an Acid Chloride
and an Amine or of a Carbamate from a Carbonochloridate and an
Amine
[0383] To a solution of an amine (1 equiv), optionally as a
hydrochloride salt, in an organic solvent (such as DCM, DCE, DMF,
DMA, NMP, THF, Et.sub.2O or 1,4-dioxane, preferably DMA, DMF or
DCM) at 0 to 25.degree. C. (preferably rt) is added a base (such as
TEA, DIEA or pyridine; 1 to 50 equiv, preferably pyridine 10 to 30
equiv or DIEA 2 to 3 equiv) and an acid chloride or a
carbonochloridate (1 to 3 equiv, preferably 1.2 equiv). The mixture
is allowed to stir at about 0 to 60.degree. C. (preferably about 25
to 50.degree. C.) for about 5 min to 20 h (preferably about 1 to 12
h). The mixture is optionally neutralized with AcOH. The mixture is
optionally concentrated in vacuo to give the final compound. The
mixture is optionally filtered through a media (such as silica gel
or Celite.RTM.) which is rinsed with an appropriate solvent (such
as EtOAc, 1,4-dioxane, THF, MeCN, DCM, Et.sub.2O, MeOH, EtOH) and
then optionally concentrated in vacuo to give a residue. Either the
residue or the solution may be optionally partitioned between water
and an organic solvent (such as EtOAc, EtO.sub.2O or DCM). The
organic layer is isolated and may be optionally washed in no
particular order with water and/or aqueous solutions containing an
acid (such as HCl, AcOH or NH.sub.4Cl) and/or aqueous solutions
containing a base (such as NaHCO.sub.3, Na.sub.2CO.sub.3, NaOH, KOH
or NH.sub.4OH) and/or aqueous solutions containing an inorganic
salt (such as NaCl Na.sub.2SO.sub.3 or NazS.sub.2O.sub.3). The
organic solution may then be optionally dried with a drying agent
(such as anhydrous MgSO.sub.4 or Na.sub.2SO.sub.4), filtered and
concentrated in vacuo to give the targeted compound.
Illustration of General Procedure F
Example #F.1:
8-((6-(1-acryloylpyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amin-
o)-4,5-dihydro-1H-benzo[b]azepin-2(3H)-one
##STR00092##
[0385] To a solution of
8-((6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-4,5-dih-
ydro-1H-benzo[b]azepin-2(3H)-one (0.090 g, 0.25 mmol, Preparation
#D.1) in DCM (4 mL) was added DIEA (0.130 mL, 0.743 mmol) followed
by acryloyl chloride (0.045 g, 0.495 mmol). The mixture was stirred
for about 12 h at about 25.degree. C. The resulting solution was
concentrated under reduced pressure to give a crude product, which
was purified by prep-HPLC (Table 1, Method i) to afford
8-((6-(1-acryloylpyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amin-
o)-4,5-dihydro-H-benzo[b]azepin-2(3H)-one (0.054 g, 52%) as a white
solid. LC/MS (Table 1, Method d) R.sub.t=2.62 min; MS m/z: 418
(M+H).sup.+. BTK enzyme IC.sub.50=A.
TABLE-US-00010 TABLE F.1 Examples prepared using an acid chloride
as described in General Procedure F R.sub.t min m/z Btk CSF-1R
Exam- (Table 1, ESI+ enzyme enzyme Amine Product ple# Method) (M +
H).sup.+ IC.sub.50 IC.sub.50 6-(3-(aminomethyl)phenyl)-
N-(bicyclo[1.1.1]pentan-1- yl)[1,2,4]triazolol[1,5-
a]pyrazin-8-amine (prepared using A from 6,8-dibromo-
[1,2,4]triazolo[1,5- a]pyrazine [Ark Pharm] and
bicyclo[1.1.1]pentan-1- amine hydrochloride [AKos], C with
(3-(4,4,5,5- tetramethyl-1,3,2- dioxaborolan-2-yl)phenyl)
methanamine hydrochloride) ##STR00093## F.1.1 2.89 (e) 361 B NT
6-(3-aminophenyl)-N- (bicyclo[1.1.1]pentan-1-yl)-
[1,2,4]triazolo[1,5-a]pyrazin- 8-amine (prepared using A from
6,8-dibromo- [1,2,4]triazolo[1,5- a]pyrazine [Ark Pharm] and
bicyclo[1.1.1] pentan-1- amine hydrochloride[AKos], C with
3-(4,4,5,5- tetramethyl-1,3,2- dioxaborolan-2-yl)aniline)
##STR00094## F.1.2 3.01 (e) 347 B NT 6-(2-aminophcnyl)-N-
(bicyclo[1.1.1]pentan-1-yl)- [1,2,4]triazolo[1,5-a]pyrazin- 8-amine
(prepared using A from 6.8-dibromo- [1,2,4]triazolo[1,5-
a]pyrazine[Ark Pharm] and bicyclo[1.1.1]pentan-1- amine
hydrochloride[AKos], C with 2-(4,4,5,5- tetramethyl-1,3,2-
dioxaborolan-2-yl)aniline) ##STR00095## F.1.3 3.08 (e) 347 C NT
8-((6-(piperidin-3-yl)- [1,2,4]triazolo[1,5-a]pyrazin-
8-yl)amino)-4,5-dihydro-1H- benzo[b]azepin-2(3H)-one (prepared
using A from 6,8- dibromo-[1,2,4]triazolo[1,5- a]pyrazine [Ark
Pharm] and 8-amino-4,5-dihydro-1H- benzo[b]azepin-2(3H)-one
[AstaTech], C with tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-
dioxaborolan-2-yl)-5,6- dihydropyridine-1(2H)- carboxylate
[Anichem], E with Pd/C, D with TFA) ##STR00096## F.1.4 2.60 (f) 432
A NT 7-((6-(3-aminophenyl)- [1,2,4]triazolo[1,5-a]pyrazin-
8-yl)amino)-4,5-dihydro-1H- benzo[b]azepin-2(3H)-one
2,2,2-trifluoroacetate (prepared using A from 6,8-
dibromo-[1,2,4]triazolo[1,5- a]pyrazine [Ark Pharm] and
8-amino-4,5-dihydro-1H- benzo[b]azepin-2(3H)-one [AstaTech], C with
3- (4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)aniline)
##STR00097## F.1.5 3.03 (d) 440 A NT 8-((6-(3-
(aminomethyl)phenyl)- [1,2,4]triazolo[1,5-a]pyrazin-
8-yl)amino)-4,5-dihydro-1H- benzo[b]azepin-2(3H)-one (prepared
using A from 6,8- dibromo-[1,2,4]triazolo[1,5- a]pyrazine [Ark
Pharm] and 8-amino-4,5-dihydro-1H- benzo[b]azepin-2(3H)-one
[AstaTech], C from (3- (4,4,3,5-tetramethyl-1,3,2- dioxaborolan-2-
yl)phenyl)methanamine hydrochloride) ##STR00098## F.1.6 2.85 (d)
454 B NT N-(6-methoxypyridazin-3- yl)-6-(pyrrolidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyridin- 8-amine (prepared using B from
8-bromo-6-chloro- [1,2,4]triazolo[1,5-a]pyridine [Example #1, Step
A] and 4- morpholinopyridin-2-amine [Matrix], C with tert-butyl 3-
(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1- carboxylate [Combi-Blocks], E with Pd/C, D
with TFA) ##STR00099## F.1.7 0.62 (d) 420 C NT
N-(3,4-dimethoxyphenyl)-6- (pyrrolidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyrazin- 8-amine (prepared using A from
6,8-dibromo- [1,2,4]triazolo[1,5- a]pyrazine [ArkPharm] and
3,4-dimethoxyaniline, C with tert-butyl 3-(4,4,5,5-
tetramethyl-1,3,2- dioxaborolan-2-yl)-2,5- dihydro-1H-pyrrole-1-
carboxylate [Combi-Blocks], E with Pd/C, D with HCl) ##STR00100##
F.1.8 2.69 (d) 395 B NT (R)-8-((6-(pyrrolidin-3-
yl)imidazo[1,2-a]pyrazin-8- yl)amino)-4,5-dihydro-1H-
benzo[b]azepin-2(3H)-one (prepared using A from 6,8-
dibromoimidazo[1,2- a]pyrazine and 8-amino-4,5-
dihydro-1H-benzo[b]azepin- 2(3H)-one [AstaTech], C with tert-butyl
3-(4,4,5,5- tetramethyl-1,3,2- dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1 - carboxylate [Combi-Blocks], E with Pd/C, G
[Table 2, Method 1], D with TFA) ##STR00101## F.1.9 2.45 (d) 417 A
NT (S)-8-((6-(pyrrolidin-3- yl)imidazo[1,2-a]pyrazin-8-
yl)amino)-4,5-dihydro-1H- benzo[b]azepin-2(3H)-one (prepared using
A from 6,8- dibromoimidazo[1,2- a]pyrazine and 8-amino-4,5-
dihydro-1H-benzo[b]azepin- 2(3H)-one [AstaTech], C with tert-butyl
3-(4,4,5,5- tetramethyl-1,3,2- dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1- carboxylate [Combi-Blocks], E with Pd/C, G
[Table 2, Method 1], D with TFA) ##STR00102## F.1.10 2.45 (d) 417 B
NT (S)-N-(3,4- dimethoxyphenyl)-6- (pyrrolidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyrazin- 8-amine (prepared using D from
Example #3, Step A with HCl) ##STR00103## F.1.11 2.40 (e) 395 B NT
(R)-N-(3,4- dimethoxyphenyl)-6- (pyrrolidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyrazin- 8-amine (prepared using D from
Example #3. Step A with HCl) ##STR00104## F.1.12 2.40 (e) 395 C NT
7-((6-(pyrrolidin-3-yl)- [1,2,4]triazolo[1,5-a]pyrazin-
8-yl)amino)-3,4- dihydroquinolin-2(1H)-one (prepared using A from
6,8- dibromo-[1,2,4]triazolo[1,5- a]pyrazine [Ark Pharm] and
7-amino-3,4- dihydroquinolin-2(1H)-one [AstaTech], C with
tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1- carboxylate [Combi-Blocks], ##STR00105##
F.1.13 2.59 (d) 404 B NT E with Pd/C, D with TFA)
6-((6-(1-acryloylpyrrolidin- 3-yl)-[1,2,4]triazolo[1,5-
a]pyrazin-8-yl)amino)-2H- benzo[b][1,4]oxazin-3(4H)- one (prepared
from 6,8-dibromo- [1,2,4]triazolo[1,5- a]pyrazine [Ark Pharm] and
6-amino-2H- benzo[b][1,4]oxazin-3(4H)- one [Bionet], C with tert-
butyl 3-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1- ##STR00106## F.1.14 2.60 (d) 406 B NT
carboxylate [Combi-Blocks], E with Pd/C, D with TFA)
8-((6-(pyrrolidin-3-yl)- [1,2,4]triazolo[1,5-a]pyridin-
8-yl)amino)-4,5-dihydro-1H- benzo[b]azepin-2(3H)-one
trifluoroacetate (prepared using B from 8-bromo-6-
chloro-[1,2,4]triazolo[1,5- a]pyridine [Example #1, Step A] and
8-amino-4,5- dihydro-1H-benzo[b]azepin- 2(3H)-one [AstaTech], C
with tert-butyl 3-(4,4,5,5- tetramethyl-1,3,2-
dioxaborolan-2-yl)-2,5- ##STR00107## F.1.15 1.76 (e) 417 B NT
dihydro-1H-pyrrole-1- carboxylate [Combi-Blocks], E with Pd/C, D
with TFA) (N-methyl-6-(pyrrolidin-3- yl)-[1,2,4]triazolo[1.5-
a]pyrazin-8-amine (prepared using A from 6,8-dibromo-
[1,2,4]triazolo[1,5- a]pyrazine [Ark Pharm] and methylamine, C with
tert- butyl 3-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1- carboxylate [Combi-Blocks], ##STR00108##
F.1.16 1.04 (f) 273 C NT E with Pd/C, D with TFA)
N-(2-methoxyethyl)-6- (pyrrolidin-3-yl)- [1,2,4]triazolo[1,5-
a]pyrazin-8-amine (prepared using A from 6,8-
dibromo-[1,2,4]triazolo[1,5- a]pyrazine [Ark Pharm] and
2-methoxyethanamine, C with tert-butyl 3-(4,4,5,5-
tetramethyl-1,3,2- dioxaborolan-2-yl)-2,5- dihydro-1H-pyrrole-1-
carboxylate [Combi-Blocks], E with Pd/C, D with TFA) ##STR00109##
F.1.17 1.50 (f) 317 C NT N-(6-morpholinopyridin-
3-yl)-6-(piperidin-3-yl)- [1,2,4]triazolo[1,5- a]pyrazin-8-amine
(prepared using A from 6,8- dibromo-[1,2,4]triazolo[1,5- a]pyrazine
[Ark Pharm] and 6-morpholinopyridin-3- amine, C with tert-butyl 3-
(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)-5,6-
dihydropyridine-1(2H)- carboxylate [Anichem], E with Pd/C, D with
TFA) ##STR00110## F.1.18 1.23 (f) 435 B NT N-(2-methoxyethyl)-6-
(pyrrolidin-3-yl)- [1,2,4]triazolo[1,5- a]pyrazin-8-amine (prepared
using A from 6,8- dibromo-[1,2,4]triazolo[1,5- a]pyrazine [Ark
Pharm] and 6-morpholinopyridin-3- amine, C with tert-butyl 3-
(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1- carboxylate [Combi-Blocks], E with Pd/C, D
with TFA) ##STR00111## F.1.19 1.16 (f) 421 B NT
N-(2-methoxyethyl)-6- (pyrrolidin-3-yl)- [1,2,4]triazolo[1,5-
a]pyrazin-8-amine (prepared using A from 6,8-
dibromo-[1,2,4]triazolo[1,5- a]pyrazine [Ark Pharm] and
1-(4-aminophenyl)-3- methylimidazolidin-2-one [Chembridge], C with
tert- butyl 3-(4,4,5,5-tetramethyl- 1,3.2-dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1- carboxylate [Combi-Blocks], E with Pd/C, D
with HCl) ##STR00112## F.1.20 1.37 (f) 433 B NT
N-(1-methyl-1H-pyrazol-3- yl)-6-(piperidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyridin- 8-amine (prepared using B from
8-bromo-6-chloro- [1,2,4]triazolo[1,5-a]pyridine [Example #1, Step
A] and 1- methyl-1H-pyrazol-3-amine, C with tert-butyl 3-(4,4.5,5-
tetramethyl-1,3,2- dioxaborolan-2-yl)-5,6- dihydropyridine-1(2H)-
carboxylate [Anichem], E with Pd/C, D with TFA) ##STR00113## F.1.21
2.56 (d) 352 B NT N-(1-methyl-1H-pyrazol-4-
yl)-6-(pyrrolidin-3-yl)- [1,2,4]triazolo[1,5-a]pyrazin- 8-amine
(prepared using A from 6,8-dibromo- [1,2,4]triazolo[1,5- a]pyrazine
[Ark Pharm] and 1-methyl-1H-pyrazol-4- amine, C with tert-butyl 3-
(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1- carboxylate [Combi-Blocks], E with Pd/C, D
with TFA) ##STR00114## F.1.22 2.38 (d) 339 A NT
N-(1-methyl-1H-pyrazol- 3-yl)-6-(pyrrolidin-3-yl)-
[1,2,4]triazolo[1,5- a]pyrazin-8-amine (prepared using A from
6,8-dibromo- [1,2,4]triazolo[1,5- a]pyrazine [ArkPharm] and
1-methyl-1H-pyrazol-3- amine, C with tert-butyl 3-
(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1- carboxylate, E with Pd(OH).sub.2/C, D with
HCl) ##STR00115## F.1.23 1.40 (h) 339 C NT
(S)-N-(6-(piperidin-3-yl)- [1,2,4]triazolo[1,5-
a]pyridin-8-yl)-6,7- dihydro-4H-pyrazolo[5,1- c][1,4]oxazin-2-amine
(prepared from Example #5, Step C, using D with HCl) ##STR00116##
F1.24 1.58 (h) 394 C NT (R)-N-(1-methyl-1H- pyrazol-4-yl)-6-
(pyrrolidin-3-yl)- [1,2,4]triazolo[1,5- a]pyrazin-8-amine (prepared
from Example #6, Step C using D with HCl) ##STR00117## F1.25 1.42
(h) 339 C NT N-(6-(pyrrolidin-3-yl)- [1,2,4]triazolo[1,5-
a]pyridin-8-yl)-6,7- dihydro-4H-pyrazolo[5,1- c][1,4]oxazin-2-amine
amine (prepared using B from tert-butyl 3-(8-
chloro-[1,2,4]triazolo[1,5- a]pyridin-6-yl)piperidine-
1-carboxylate [prepared using C with 6-bromo-8-
chloro-[1,2,4]triazolo[1,5- a]pyridine [Example #1, Step A] and
tert-butyl 3- (4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1- ##STR00118## F1.26 0.66 (a) 380 A NT
carboxylate [Cobmi-Blocks], E with Pt/C] and 6,7-
dihydro-4H-pyrazolo[5,1- c][1,4]oxazin-2-amine [Preparation #1], D
with TFA) N-(6-morpholinopyridazin-3- yl)-6-(piperidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyridin- 8-amine (prepared using B from
tert-butyl 3-(8- chloro-[1,2,4]triazolo[1,5-
a]pyridin-6-yl)piperidine- 1-carboxylate [prepared using C with
6-bromo-8- chloro-[1,2,4]triazolo[1,5- a]pyridine [Example #1, Step
A] and tert-butyl 3- (4,4,5,5-tetramethyl-1,3,2-
dioxaborolan-2-yl)-5,6- dihydropyridine-1(2H)- carboxylate
[Anichem], E ##STR00119## F1.27 1.13(e) 435 NT NT with Pt/C] and
3-amino-6- (morpholin-4-yl)pyridazine (Matrix), D with TFA)
N-(5-morpholinopyridin-2- yl)-6-(piperidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyridin- 8-amine (prepared using B from
tert-butyl 3-(8- chloro-[1,2,4]triazolo[1,5-
a]pyridin-6-yl)piperidine- 1-carboxylate [prepared using C with
6-bromo-8- chloro-[1,2,4]triazolo[1,5- a]pyridine [Example #1, Step
A] and tert-butyl 3- (4,4,5,5-tetramethyl-1,3,2-
dioxaborolan-2-yl)-5,6- dihydropyridine-1(2H)- carboxylate
[Anichem], E ##STR00120## F1.28 110 (a) 434 B NT with Pt/C] and 5-
morpholinopyridin-2-amine (Oakwood), D with TFA)
N-(6-morpholinopyridin-2- yl)-6-(piperidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyridin- 8-amine (prepared using B from
tert-butyl 3-(8- chloro-[1,2,4]triazolo[1,5-
a]pyridin-6-yl)piperidine- 1-carboxylate [prepared using C with
6-bromo-8- chloro-[1,2,4]triazolo[1,5- a]pyridine [Example #1 Step
A] and tert-butyl 3-(4,4,5,5- tetramethyl-1,3,2-
dioxaborolan-2-yl)-5,6- dihydropyridine-1(2H)- carboxylate
[Anichem], E with Pt/C] and 6- morpholinopyridin-2-amine
(Combi-Phos), D with TFA) ##STR00121## F1.29 2.02 (e) 434 B NT
N-(6-piperidin-3-yl)- [1,2,4]triazolo[1,5- a]pyridin-8-yl)-6,7-
dihydro-4H-pyrazolo[5,1- c][1,4]oxazin-2-amine (prepared using B
from tert- butyl 3-(8-chloro- [1,2,4]triazolo[1,5-
a]pyridin-6-yl)piperidine- 1-carboxylate [prepared using C with
6-bromo-8- chloro-[1,2,4]triazolo[1,5- a]pyridine [Example #1, Step
A] and tert-butyl 3- (4,4,5,5-tetramethyl-1,3,2- ##STR00122## F1.30
1.75 (e) 394 B NT dioxaborolan-2-yl)-5,6- dihydropyridine-1(2H)-
carboxylate [Anichem], E with Pt/C] and 6,7-dihydro-
4H-pyrazolo[5,1- c][1,4]oxazin-2-amine [Preparation #1], D with
TFA) 3-isopropyl-N-(6-(piperidin- 3-yl)-[1,2,4]triazolo[1,5-
a]pyridin-8-yl)-1,2,4- oxadiazol-5-amine (prepared using B from
tert-butyl 3- (8-chloro- [1,2,4]triazolo[1,5-
a]pyridin-6-yl)piperidine- 1-carboxylate [prepared using C with
6-bromo-8- chloro-[1,2,4]triazolo[1,5- a]pyridine [Example #1, Step
A] and tert-butyl 3- (4,4,5,5-tetramethyl-1,3,2-
dioxaborolan-2-yl)-5,6- dihydropyridine-1(2H)- ##STR00123## F1.31
1.95 (e) 382 C NT carboxylate [Anichem], E with Pt/C] and
3-isopropyl- 1,2,4-oxadiazol-5-amine (Ark Pharm), D with TFA)
3-methyl-(N-(6-(piperidin-3- yl)-[1,2,4]triazolo[1,5-
a]pyridin-8-yl)-1,2,4- oxadiazol-5-amine (prepared using B from
tert-butyl 3- (8-chloro- [1,2,4]triazolo[1,5-
a]pyridin-6-yl)piperidine- 1-carboxylate [prepared using C with
6-bromo-8- chloro-[1,2,4]triazolo[1,5- a]pyridine [Example #1, Step
A] and tert-butyl 3- (4,4,5,5-tetramethyl-1,3,2-
dioxaborolan-2-yl)-5,6- ##STR00124## F1.32 1.27 (e) 354 C NT
dihydropyridine-1(2H)- carboxylate [Anichem], E with Pt/C] and
3-methyl- 1,2,4-oxadiazol-5- amine(Matrix), D with TFA)
(S)-N-(6-(pyrrolidin-3-yl)- [1,2,4]triazolo[1,5-
a]pyridin-8-yl)-6,7- dihydro-4H-pyrazolo[5,1- c][1,4]oxazin-2-amine
(prepared using B from- [1,2,4]triazolo[1,5- a]pyridine [Example
#1, Step A] and 6,7-dihydro- 4H-pyrazolo[5,1- c][1,4]oxazin-2-amine
[Preparation #1], C with tert-butyl 3-(4,4,5,5- tetramethyl-1,3,2-
dioxaborolan-2-yl)-2,5- dihydro-1H-pyrrole-1- carboxylate [Combi-
Blocks], E with Pd/C, G ##STR00125## F1.33 1.42 (l) 380 C NT [Table
2, Method 7], D with HCl) 6-((1R,3S)-3- aminocyclopentyl)-N-(1-
methyl-1H-pyrazol-3-yl)- [1,2,4]triazolo[1,5- a]pyridin-8-amine
hydrochloride (prepared using Example #7, Step E using (2S,5S)-5-
benzyl-3-methyl-2-(5- methylfuran-2- yl)imidazolidin-4-one, then
Example #7, Step F, D with HCl) ##STR00126## F1.34 1.49 (a) 352 B
NT 6-(cis-3-aminocyclohexyl)- N-(1-methyl-1H-pyrazol-3-
yl)-[1,2,4]triazolo[1,5- a]pyridin-8-amine (prepared using B from
8-bromo-6- chloro-[1,2,4]triazolo[1,5- a]pyridine [Example #1, Step
A] and 1-methyl-1H- pyrazol-3-amine, C with [3- (4,4,5,5-
tetramethyl[1,3,2]dioxaborolan- 2-yl)-cyclohex-3-enyl]- carbamic
acid tert-butyl ester and [3-(4,4,5,5-tetramethyl- ##STR00127##
F1.35 2.62 (b) 366 A NT [1,3,2]dioxaborolan-2-yl)-
cyclohex-2-enyl]-carbamic acid tert-butyl ester [prepared according
to U.S. 2009/0197864]. E with Pd/C, D with HCl)
6-(t-3-aminocyclohexyl)-N- (1-methyl-1H-pyrazol-3-yl)-
[1,2,4]triazolo[1,5-a]pyridin- 8-amine (prepared using B from
8-bromo-6-chloro- [1,2,4]triazolo[1,5-a]pyridine [Example #1, Step
A] and 1- methyl-1H-pyrazol-3-amine, C with [3-(4,4,5,5-
tetramethyl [1,3,2] dioxaborolan- 2-yl)-cyclohex-3-enyl]- carbamic
acid tert-butyl ester and [3-(4,4,5,5-tetramethyl-
[1,3,2]dioxaborolan-2-yl)- ##STR00128## F1.36 2.64 (d) 366 B NT
cyclohex-2-enyl]-carbamic acid tert-butyl ester [prepared according
to U.S. 2009/0197864], E with Pd/C, D with HCl)
(S)-N-(1-methyl-1H-pyrazol- 3-yl)-6-(pyrrolidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyridin- 8-amine (prepared using C from
tert-butyl 3-(4,4,5,5- tetramethyl-1,3,2- dioxaborolan-2-yl)-2,5-
dihydro-1H-pyrrole-1- carboxylate [Combi-Blocks] and Preparation
#B.1, E with Pd/C, G [Table 2, Method 10], D with HCl) ##STR00129##
F1.37 1.43 (h) 338 C NT (R)-N-(1-methyl-1H-pyrazol-
3-yl)-6-(pyrrolidin-3-yl)- [1,2,4]triazolo[1,5-a]pyridin- 8-amine
(prepared using C from tert-butyl 3-(4,4,5,5- tetramethyl-1,3,2-
dioxaborolan-2-yl)-2,5- dihydro-1H-pyrrole-1- carboxylate
[Combi-Blocks] and Preparation #B.1, E with Pd/C, G [Table 2,
Method 10], D with HCl) ##STR00130## F.1.38 1.43 (h) 338 A NT
(S)-N-(5-methyl-4,5,6,7- tetrahydropyrazolo[1,5- a]pyrazin-2-yl)-6-
(pyrrolidin-3-yl)- [1,2,4]triazolo[1,5-a]pyridin- 8-amine
hydrochloride (prepared using B from Example #1, Step A and 5-
methyl-4,5,6,7- tetrahydropyrazolo[1,5- a]pyrazin-2-amine
[Astatech], C with tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-
dioxaborolan-2-yl)-2,5- dihydro-1H-pyrrole-1- carboxylate
[Combi-Blocks], E with Pd/C, G [Table 2, ##STR00131## F.1.39 1.37
(m) 393 A NT Method 11], D) (S)-N-(5-methyl-4,5,6,7-
tetrahydropyrazolo[1,5- a]pyrazin-2-yl)-6-(piperidin-
3-yl)-[1,2,4]triazolo[1,5- a]pyridin-8-amine hydrochloride
(prepared using B from Example #1, Step A and 5-methyl-4,5,6,7-
tetrahydropyrazolo[1,5- a]pyrazin-2-amine [Astatech], C with
tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)-5,6-
dihydropyridine-1(2H)- carboxylate [Anichem], E with Pd(OH)2, G
[Table 2, ##STR00132## F.1.40 1.49 (n) 407 A NT Method 12], D)
N-(6-(pyrrolidin-3- yl)imidazo[1,2-b]pyridazin-
8-yl)-6,7-dihydro-4H- pyrazolo[5,1-c][1,4]oxazin- 2-amine,
hydrochloric acid (prepared using B from 8-bromo-6-
chloroimidazo[1,2- b]pyridazine, hydrochloric acid [Astatech] and
Preparation #1, C with tert- butyl 3-(4,4,5,5-tetramethyl-
1,3,2-dioxaborolan-2-yl)-2,5- dihydro-1H-pyrrole-1- carboxylate
[Combi-Blocks], ##STR00133## F.1.41 1.47 (a) 380 A NT E with
Pd(OH).sub.2, D) N-(6-(azetidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyridin- 8-yl)-6,7-dihydro-4H-
pyrazolo[5,1-c][1,4]oxazin- 2-amine hydrochloride (prepared using D
from Preparation #2) ##STR00134## F.1.42 1.34 (o) 366 A NT
6-cyclohexyl-N-(1- (piperidin-4-yl)-1H-pyrazol-
4-yl)-[1,2,4]triazolo[1,5- a]pyrazin-8-amine hydrochloride
(prepared using A from 6,8-dibromo- [1,2,4]triazolo[1,5- a]pyrazine
[ArkPharm] and tert-butyl 4-(4-amino-1H- pyrazol-1-yl)piperidine-1-
carboxylate [Combiblocks], C with 2-(cyclohex-1-en-1-yl)-
4,4,5,5-tetramethyl-1,3,2- dioxaborolane [Combiblocks], E with
Pd(OH).sub.2/C, and D with HC1. ##STR00135## F.1.43 2.00 (h) 409 NT
A 6-cyclohexyl-N-(1- (piperidin-4-yl)-1H-pyrazol-
4-yl)-[1,2,4]triazolo[1,5- a]pyrazin-8-amine hydrochloride
(prepared using A from 6,8-dibromo- [1,2,4]triazolo[1,5- a]pyrazine
[ArkPharm] and tert-butyl 4-(4-amino-1H- pyrazol-1-yl)piperidine-1-
carboxylate [Combiblocks], C with 2-(cyclohex-1-en-1-yl)-
4,4,5,5-tetramethyl-1,3,2- dioxaborolane [Combiblocks], E with
Pd(OH).sub.2/C, and D with HCl. ##STR00136## F1.44 1.99 (h) 424 NT
A
General Procedure G: Chiral Preparative HPLC Separation of
Stereoisomers
[0386] A racemic mixture of compound is subjected to chiral
purification using preparative HPLC. The representative gradients
using mobile phases A and B are described in Table 2, Methods
(1-6). When indicated, methods (1-6) from Table 2, were employed
respectively for the chiral separation of the racemic mixtures.
Illustration of General Procedure G
Preparation #G.1 and #G.2: (S)-tert-butyl
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)pyrrolidine-1-carboxylate and (R)-tert-butyl
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)pyrrolidine-1-carboxylate
##STR00137##
[0388] A racemic mixture of tert-butyl
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)pyrrolidine-1-carboxylate (Example #6, step C) was separated via
chiral prep (Table 2, Method 6) to give (S)-tert-butyl
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)pyrrolidine-1-carboxylate (0.435 g, 40.2%, OR=negative) and
(R)-tert-butyl
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)pyrrolidine-1-carboxylate (0.442 g, 40.9%, OR=positive)
[Stereochemistry is arbitrarily assigned]: LC/MS (Table 1, Method
1) R.sub.t=1.96 min.; MS m/z: 385 (M+H).sup.+.
TABLE-US-00011 TABLE G.1 Examples prepared using chiral SFC (Table
2, Method 17 or 18) from General Procedure G R.sub.t min R.sub.t
min m/z CSF-1R Exam- (Table 1, (Table 2, ESI+ enzyme Racemate
Products ple # Method) Method) (M + H).sup.+ IC.sub.50
(trans)-3-(4-((6-cyclohexyl- [1,2,4]triazolo[1,5-
a]pyrazin-8-yl)amino)-1H pyrazol-1-yl)cyclohexanol (prepared using
A from 8- bromo-6-cyclohexyl- [1,2,4]triazolo[1,5- a]pyrazine
[Example #11, Step E] and (trans)-3-(4- amino-1H-pyrazol-1-
yl)cyclohexanol [Preparation #7] ##STR00138## G.1.1 1.45 (v) 1.86
(17) 382 A (trans)-3-(4-((6-cyclohexyl- [1,2,4]triazolo[1,5-
a]pyrazin-8-yl)amino)-1H- pyrazol-1-yl)cyclohexanol (prepared using
A from 8- bromo-6-cyclohexyl- [1,2,4]triazolo[1,5- a]pyrazine
[Example #11, Step D] and (trans)-3-(4- amino-1H-pyrazol-1-
yl)cyclohexanol [Preparation #7] ##STR00139## G.1.2 3.23 (v) 1.98
(17) 382 A (cis)-3-(4-((6-cyclohexyl- [1,2,4]triazolo[1,5-
a]pyrazin-8-yl)amino)-1H- pyrazol-1-yl)cyclohexanol (prepared using
A from 8- bromo-6-cyclohexyl- [1,2,4]triazolo[1,5- a]pyrazine
[Example #11, Step D] and (cis)-3-(4- amino-1H-pyrazol-1-
yl)cyclohexanol [Preparation #8] ##STR00140## G.1.3 3.23 (v) 2.10
(18) 382 A (cis)-3-(4-((6-cyclohexyl- [1,2,4]triazolo[1,5-
a]pyrazin-8-yl)amino)-1H- pyrazol-1-yl)cyclohexanol (prepared using
A from 8- bromo-6-cyclohexyl- [1,2,4]triazolo[1,5- a]pyrazine
[Example #11, Step D] and (cis)-3-(4- amino-1H-pyrazol-1-
yl)cyclohexanol [Preparation #8] ##STR00141## G.1.4 3.13 (v) 2.29
(18) 382 A
General Procedure H: Formation of a Sulfonamide from a Sulfonyl
Chloride and an Amine
[0389] To a solution of an amine (preferably 1 equiv) and a base
(such as pyridine, DIEA or TEA; 1.1-5 equiv, preferably 2-3 equiv
DIEA) in an organic solvent (for example, DMA, DMF or THF,
preferably DMA) at about 0.degree. C. is added a sulfonyl chloride
(1.0-1.2 equiv). After about 5-120 min (preferably 10-30 min), the
reaction mixture is added slowly into ice water while stirring. If
a precipitate is present after the ice melted completely, the
resulting solid is collected via vacuum filtration to give crude
product. Alternatively an extractive work up or concentration of
the reaction mixture under reduced pressure may be done.
Illustration of General Procedure H
Example #H.1:
(S)--N-(3,4-dimethoxyphenyl)-6-(1-(vinylsulfonyl)pyrrolidin-3-yl)-[1,2,4]-
triazolo[1,5-a]pyrazin-8-amine
##STR00142##
[0391] To a solution of
(S)--N-(3,4-dimethoxyphenyl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]py-
razin-8-amine (0.073 g, 0.21 mmol, Example #3, Step B) and DIEA
(0.093 mL, 0.54 mmol) in DMA (1.5 mL) at about 0.degree. C. was
added ethenesulfonyl chloride (0.025 mL, 0.22 mmol). After about 20
min, the reaction mixture was added slowly into ice water
(.about.10 mL) while stirring. After the ice melted completely, the
resulting solid was collected via vacuum filtration and dried under
vacuum at 55.degree. C. to give impure product. The solid was
triturated with MeOH (1 mL). The resulting solid was collected via
vacuum filtration while washing with additional MeOH (1 mL) and
dried under vacuum at 55.degree. C. to give solid with little
change in purity by LCMS. The filtrate and solid were recombined,
concentrated under reduced pressure and purified via silica gel
chromatography eluting with 0-5% MeOH in DCM. The
product-containing fraction was concentrated under reduced pressure
and dried under vacuum at about 55.degree. C. to give
(S)--N-(3,4-dimethoxyphenyl)-6-(1-(vinylsulfonyl)pyrrolidin-3-yl)-[1,2,4]-
triazolo[1,5-a]pyrazin-8-amine (0.024 g, 26%) as a white solid:
LC/MS (Table 1, Method e) R.sub.t=2.00 min.; MS m/z: 431
(M+H).sup.+. BTK enzyme IC.sub.50=A
General Procedure I: Formation of a Cyanamide from an Amine with
Cyanogen Bromide
[0392] To a mixture of an amine (preferably 1 equiv) and a base
(such as cesium carbonate or TEA; 1-10 equiv, preferably 1-5 equiv)
in a solvent such as DCM, DMA, THF or DMF (preferably DMA or THF)
at about -5 to 25.degree. C. (preferably 0.degree. C.) is added
cyanogen bromide (1-3 equivalents, preferably 1.1-2.2 equiv). The
reaction temperature is maintained or is allowed to warm. After
about 5-120 min, the reaction mixture is diluted with water (10 mL)
and extracted (for example with DCM). The combined organic layers
is optionally washed with brine, dried over Na.sub.2SO.sub.4 or
MgSO.sub.4, filtered, and concentrated under reduced pressure.
Illustration of General Procedure I
Example #I.1:
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)pyrrolidine-1-carbonitrile
##STR00143##
[0394] To a mixture of
N-(1-methyl-1H-pyrazol-3-yl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]py-
ridin-8-amine (0.10 g, 0.35 mmol) and cesium carbonate (0.460 g,
1.41 mmol) in DMA (3.0 mL) at about 0.degree. C. was added cyanogen
bromide (0.075 g, 0.71 mmol). The reaction bath was allowed to warm
to about 10.degree. C. After about 30 min, diluted with water (10
mL) and extracted with DCM (3.times.15 mL). The combined organic
layers were washed with brine (15 mL), dried over MgSO.sub.4,
filtered, and concentrated under reduced pressure. The residue was
dissolved in DCM for purification via silica gel chromatography
eluting with 0-50% DCM/MeOH/NH.sub.4OH (90:9:1) in DCM to give
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)pyrrolidine-1-carbonitrile (0.076 g, 70%) as a pale yellow solid
after drying under vacuum at about 55.degree. C. LC/MS (Table 1,
Method e) R.sub.t=1.43 min.; MS m/z: 309 (M+H).sup.+. BTK enzyme
IC.sub.50=B
TABLE-US-00012 TABLE 1.1 Examples prepared from cyanogen bromide
using General Procedure I R.sub.t min m/z BTK Exam- (Table 1, ESI+
enzyme Amine Product ple # Method) (M + H).sup.+ IC.sub.50
N-(1-methyl-1H-pyrazol-3-yl)-6- (piperidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyridin-8- amine (prepared using B from 8-
bromo-6-chloro- [1,2,4]triazolo[1,5-a]pyridine [Example #1, Step A]
and 1- methyl-1H-pyrazol-3-amine, C with tert-butyl 3-(4,4,5,5-
tetramethyl-1,3,2-dioxaborolan- 2-yl)-5,6-dihydropyridine-1(2H)-
carboxylatc [Anichem], E with Pd/C, D with TFA) ##STR00144## I.1.1
1.63 (e) 323 B N-(1-methyl-1H-pyrazol-4-yl)-6- (pyrrolidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyrazin-8- amine (prepared from 6,8-
dibromo-[1,2,4]triazolo[1,5- a]pyrazine [Ark Pharm] and 1-
methyl-1H-pyrazol-4-amine, C with tert-butyl 3-(4,4,5,5-
tetramethyl-1,3,2-dioxaborolan- 2-yl)-2,5-dihydro-1H-pyrrole-1-
carboxylate [Combi-Blocks], E with Pd/C, D with TFA) ##STR00145##
I.1.2 1.44 (e) 310 B N-(6-(piperidin-3-yl)-
[1,2,4]triazolo[1,5-a]pyridin- 8-yl)-6,7-dihydro-4H-
pyrazolo[5,1-c][1,4]oxazin-2- amine (prepared using B from
tert-butyl 3-(8-chloro- [1,2,4]triazolo[1,5-a]pyridin-
6-yl)piperidine-1-carboxylate [prepared using C with 6-
bromo-8-chloro- [1,2,4]triazolo[1,5-a]pyridine [Example #1, Step A]
and tert- butyl 3-(4,4,5,5-tetramethyl-
1,3,2-dioxaborolan-2-yl)-5,6- dihydropyridine-1(2H)- ##STR00146##
I.1.3 1.66 (e) 365 B carboxylate [Anichem], E with Pt/C] and
6,7-dihydro-4H- pyrazolo[5,1-c][1,4]oxazin-2- amine [Preparation
#1] D with TFA) 6-(trans-3-aminocyclohexyl)-N-
(1-methyl-1H-pyrazol-3-yl)- [1,2,4]triazolo[1,5-a]pyridin-8- amine
(prepared using B from 8- bromo-6-chloro-
[1,2,4]triazolo[1,5-a]pyridine [Example #1, Step A] and 1-
methyl-1H-pyrazol-3-amine, C with [3-(4,4,5,5-
tetramethyl[1,3,2]dioxaborolan-2- yl)-cyclohex-3-enyl]-carbamic
acid tert-butyl ester and [3- (4,4,5,5-tetramethyl-
[1,3,2]dioxaborolan-2-yl)- ##STR00147## 1.1.4 2.48 (f) 337 B
cyclohex-2-enyl]-carbamic acid tert-butyl ester [prepared according
to U.S. 2009/0197864], E with Pd/C, D with HCl)
Example #1.
1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)pyrrolidin-1-yl)prop-2-en-1-one
##STR00148##
[0395] Step A: 8-bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine
##STR00149##
[0397] To a solution of 2-amino-3-bromo-5-chloropyridine (10.0 g,
48.2 mmol, Ark Pharm) in N,N-dimethylformamide (20 mL) was added
DMF-DMA (17.2 g, 145 mmol) and the mixture was stirred at
130.degree. C. for about 18 h. The mixture was cooled and
evaporated to dryness. To an ice cooled stirring solution of the
brown solid in MeOH (80.0 mL) and pyridine (7.80 mL, 96 mmol) was
added hydroxylamine-o-sulfonic acid (7.63 g, 67.5 mmol). The
reaction was allowed to warm to about 25.degree. C. and stirred for
about 18 h. The mixture was evaporated and the solid residue was
dissolved in DCM (150 mL) and washed with saturated sodium
bicarbonate (200 mL), water (200 mL) and brine (200 mL). The
organic mixture was filtered through a Biotage phase separator to
remove residual water and evaporated to dryness to give
8-bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine as an orange solid,
which was used in the next step without further purification. (6.1
g, 64% crude): .sup.1H NMR (CDCl.sub.3) .delta. 8.65 (d, J=1.8 Hz,
1H), 8.39 (s, 1H), 7.80 (d, J=1.7 Hz, 1H).
Step B.
6-chloro-N-(1-methyl-1H-pyrazol-3-yl)-[1,2,4]triazolo[1,5-a]pyridi-
n-8-amine
##STR00150##
[0399] To a microwave vial was added
8-bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine (1.59 g, 6.84 mmol,
Example #1, Step A), 1-methyl-1H-pyrazol-3-amine (0.731 g, 7.52
mmol, Matrix Scientific), Xantphos (0.831 g, 1.44 mmol),
palladium(II) acetate (0.154 g, 0.684 mmol), cesium carbonate (5.57
g, 17.1 mmol) and 1,4-dioxane (12 mL). The mixture was then heated
in a microwave for about 1.5 h at about 120.degree. C. The reaction
was cooled to rt and concentrated under reduced pressure. The
residue was sonicated with MeOH (10 mL) to give a uniform
suspension, followed by filtration. LC/MS indicated impure product
in both solid and filtrate. The solid and filtrate were combined,
concentrated under reduced pressure, redissolved in DCM/MeOH (3:1;
250 mL), followed by addition of Silica gel. The suspension was
concentrated under reduced pressure, purified via silica gel
chromatography eluting with 0-60% DCM/MeOH/NH.sub.4OH (90:9:1) in
DCM. The product-containing fractions were combined and
concentrated under reduced pressure to give
6-chloro-N-(1-methyl-1H-pyrazol-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-ami-
ne (1.63 g, .about.96%, .about.86% purity by NMR): LC/MS (Table 1,
Method e) R.sub.t=2.33 min.; MS m/z: 248 (M+H).sup.+.
Step C. tert-butyl
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)-2,5-dihydro-1H-pyrrole-1-carboxylate
##STR00151##
[0401] To a 10 mL microwave tube were added
6-chloro-N-(1-methyl-1H-pyrazol-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-ami-
ne (0.21 g, 0.84 mmol), tert-butyl
3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro-1H-pyrrole-1--
carboxylate (0.35 g, 1.2 mmol, Combi-Blocks), THF (5 mL), MeOH (1
mL) and sodium carbonate (1.27 mL, 2.53 mmol). The mixture was
sparged with nitrogen, then
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex (0.069 g, 0.084 mmol) was added. The vial
was sealed. The mixture was heated in a microwave at about
130.degree. C. for about 1 h. The reaction mixture was filtered
through Celite.RTM., washed with DCM and MeOH. The filtrate was
concentrated under reduced pressure. The crude product was purified
via silica gel chromatography eluting with 0-80%
DCM/MeOH/NH.sub.4OH (90:9:1) in DCM. The product-containing
fractions were combined, concentrated under reduced pressure,
triturated with MeOH (1 mL). The resulting solid was collected via
vacuum filtration, washed with additional MeOH (5 mL) and then
dried under vacuum at about 55.degree. C. to give tert-butyl
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)-2,5-dihydro-1H-pyrrole-1-carboxylate (0.17 g, 53%): LC/MS (Table
1, Method e) R.sub.t=3.09 min.; MS m/z: 382 (M+H).sup.+.
Step D. tert-butyl
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)pyrrolidine-1-carboxylate
##STR00152##
[0403] tert-Butyl
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)-2,5-dihydro-1H-pyrrole-1-carboxylate (1.40 g, 3.67 mmol), MeOH
(100 mL), THF (5 mL), AcOH (5 mL) were added to dry 10% Pd/C (0.450
g, 0.423 mmol) in a 250 mL stainless steel pressure bottle. The
reaction mixture was stirred at rt for about 16 h with 30 psi of
H.sub.2. Analytical HPLC indicated starting material and product
present. Dry 10% Pd/C (0.450 g, 0.423 mmol) was added and
hydrogenation continued at rt for about 4 days. The mixture was
filtered through a nylon membrane and concentrated under reduced
pressure. The residue was purified via silica gel chromatography
eluting with 0-50% DCM/MeOH/NH.sub.4OH (90:9:1) in DCM. The
product-containing fractions were combined and concentrated under
reduced pressure and dried under vacuum at about 55.degree. C. to
give tert-butyl
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)pyrrolidine-1-carboxylate (0.752 g, 52%): LC/MS (Table 1, Method
e) R.sub.t=2.21 min.; MS m/z: 384 (M+H).sup.+.
Step E.
N-(1-methyl-1H-pyrazol-3-yl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1-
,5-a]pyridin-8-amine
##STR00153##
[0405] To a solution of tert-butyl
3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)pyrrolidine-1-carboxylate (0.076 g, 0.20 mmol) in MeOH (0.75 mL)
was added HCl (4M in 1,4-dioxane, 0.75 mL, 3.00 mmol). The reaction
was stirred at rt for about 3.5 h and then concentrated under
reduced pressure. The residue was purified via silica gel
chromatography eluting with 0-100% DCM/MeOH/NH.sub.4OH (90:9:1) in
DCM. The product-containing fractions were combined, concentrated
under reduced pressure and dried under vacuum at about 55.degree.
C. to give
N-(1-methyl-1H-pyrazol-3-yl)-6-(pyrrolidin-3-yl)-1,2,4/triazolo[1,5-a]pyr-
idin-8-amine (0.061 g, .about.109%, .about.91% by NMR) which was
used in the next step without further purification: LC/MS (Table 1,
Method e) R.sub.t=0.63 min.; MS m/z: 284 (M+H).sup.+.
Step F.
1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]py-
ridin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one
##STR00154##
[0407] To a solution of
N-(1-methyl-1H-pyrazol-3-yl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]py-
ridin-8-amine (0.143 g, 0.505 mmol) and
N-ethyl-N-isopropylpropan-2-amine (0.220 mL, 1.26 mmol) in DMA (2.5
mL) at about 0.degree. C. was added acryloyl chloride (0.043 mL,
0.530 mmol). After about 2 min, the ice bath was removed and the
reaction was allowed to stir at rt. After about 25 min, the
reaction mixture was added diluted with water (10 mL), extracted
with DCM (3.times.10 mL). The combined organic layers were washed
with brine, dried over MgSO.sub.4, filtered, and concentrated under
reduced pressure. The crude product was purified via silica gel
chromatography eluting with 50-100% EtOAc in DCM with long hold at
100% EtOAc. The product-containing fractions were combined,
concentrated under reduced pressure, and dried under vacuum at
about 55.degree. C. to give
1-(3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)pyrrolidin-1-yl)prop-2-en-1-one (0.11 g, 65%) as a white foam:
LC/MS (Table 1, Method e) R.sub.t=1.56 min.; MS m/z: 338
(M+H).sup.+. BTK enzyme IC.sub.50=A
Example #2:
1-(3-(8-(6-Methoxypyridazin-3-ylamino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)pyrrolidin-1-yl)prop-2-en-1-one trifluoroacetate
##STR00155##
[0408] Step A:
6-chloro-N-(6-methoxypyridazin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-amin-
e
##STR00156##
[0410] To a microwave tube were added
8-bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine (1.00 g, 4.30 mmol,
Example #1, Step A), 6-methoxypyridazin-3-amine (0.538 g, 4.30
mmol), Xantphos (0.523 g, 0.903 mmol), Pd(OAc).sub.2 (0.097 g, 0.43
mmol), cesium carbonate (2.80 g, 8.60 mmol) and 1,4-dioxane (15
mL). The reaction was degassed with vacuum and back filled with
nitrogen. The vessel was sealed and heated for about 1.5 h at about
120.degree. C. in a microwave oven. The mixture was cooled,
filtered through Celite.RTM. and concentrated. The resulting
residue was purified via silica gel chromatography eluting with a
gradient of 20-70% EtOAc in hexane to give
6-chloro-N-(6-methoxypyridazin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-amin-
e (0.500 g, 42%); .sup.1H NMR (CDCl.sub.3) .delta. 8.84 (d, J=1.7
Hz, 1H), 8.30-8.20 (m, 2H), 7.65 (s, 1H), 7.09 (t, J=6.8 Hz, 1H),
7.00 (dd, J=12.4, 6.7 Hz, 1H), 4.12 (d, J=7.1 Hz, 3H).
Step B: tert-butyl
3-(8-(6-methoxypyridazin-3-ylamino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-2-
,5-dihydro-1H-pyrrole-1-carboxylate
##STR00157##
[0412] To a mixture of
6-chloro-N-(6-methoxypyridazin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-amin-
e (3.45 g, 12.47 mmol), tert-butyl
3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro-1H-pyrrole-1--
carboxylate (5.52 g, 18.7 mmol, Combi-Blocks), potassium carbonate
(3.45 g, 24.9 mmol) 1,4-dioxane (100 mL), and water (50 mL) were
added Pd.sub.2(dba).sub.3 (1.14 g, 1.25 mmol) and
dicyclohexyl(2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphine
(0.594 g, 1.25 mmol) under nitrogen. The mixture was heated at
about 100.degree. C. under nitrogen for about 16 h. The reaction
mixture was partitioned between water and DCM. The organic layer
was concentrated and purified via silica gel chromatography eluting
with a gradient of 20-80% EtOAc in hexane to give tert-butyl
3-(8-((6-methoxypyridazin-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-
-2,5-dihydro-1H-pyrrole-1-carboxylate (3.2 g, 62%) as a white
solid. LC/MS (Table 1, Method d) R.sub.t=1.38 min; MS m/z: 410
(M+H).sup.+.
Step C: tert-butyl
3-(8-(6-methoxypyridazin-3-ylamino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)py-
rrolidine-1-carboxylate
##STR00158##
[0414] To a solution of tert-butyl
3-(8-(6-methoxypyridazin-3-ylamino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-2-
,5-dihydro-1H-pyrrole-1-carboxylate (0.20 g, 0.49 mmol) in THF (120
mL) was added 10% Pd/C (0.50 g, 0.47 mmol). The mixture was stirred
at rt under about 25 psi of H.sub.2 for about 24 h. The mixture was
filtered and concentrated under reduced pressure to give tert-butyl
3-(8-((6-methoxypyridazin-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-
pyrrolidine-1-carboxylate (0.18 g, 90%). LC/MS (Table 1, Method d)
R.sub.t=1.01 min; MS m/z: 412 (M+H).sup.+.
Step D:
N-(6-methoxypyridazin-3-yl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,-
5-a]pyridin-8-amine
##STR00159##
[0416] To a solution of tert-butyl
3-(8-((6-methoxypyridazin-3-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-
pyrrolidine-1-carboxylate (0.600 g, 1.46 mmol) in DCM (360 mL) was
added TFA (12 mL, 156 mmol). The mixture was stirred at rt for
about 6 h. The mixture was concentrated to give
N-(6-methoxypyridazin-3-yl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyr-
idin-8-amine which was used in next step without further
purification. (0.50 g, 100% crude); LC/MS (Table 1, Method d)
R.sub.t=1.46 min; MS nm/z: 312 (M+H).sup.+.
Step E:
1-(3-(8-(6-methoxypyridazin-3-ylamino)-[1,2,4]triazolo[1,5-a]pyrid-
in-6-yl)pyrrolidin-1-yl)prop-2-en-1-one trifluoroacetate
##STR00160##
[0418] To a solution of
N-(6-methoxypyridazin-3-yl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]pyr-
idin-8-amine (0.080 g, 0.26 mmol) and TEA (0.143 mL, 1.03 mmol) in
DCM (2 mL) was added acryloyl chloride (0.023 g, 0.257 mmol) at
about 0.degree. C. The mixture was stirred at rt overnight. Water
was added to quench the reaction. The reaction mixture was
partitioned between water and DCM. The organic layer was
concentrated and purified by prep-HPLC (Table 1, Method z) to give
1-(3-(8-(6-methoxypyridazin-3-ylamino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl-
)pyrrolidin-1-yl)prop-2-en-1-one trifluoroacetate. (0.049 g, 52%);
LC/MS (Table 1, Method d) R.sub.t=2.29 min; MS m/z: 366
(M+H).sup.+. BTK enzyme IC.sub.50=B
Example #3.
(S)--N-(3,4-dimethoxyphenyl)-6-(1-(vinylsulfonyl)pyrrolidin-3-yl)-[1,2,4]-
triazolo[1,5-a]pyrazin-8-amine
##STR00161##
[0419] Step A. (S)-tert-butyl
3-(8-((3,4-dimethoxyphenyl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)pyrr-
olidine-1-carboxylate and (R)-tert-butyl
3-(8-((3,4-dimethoxyphenyl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)pyrr-
olidine-1-carboxylate
##STR00162##
[0421] To dry 10% Pd/C (0.50 g, 0.40 mmol) in a 250 mL stainless
steel pressure bottle was added tert-butyl
3-(8-((3,4-dimethoxyphenyl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)-2,5-
-dihydro-1H-pyrrole-1-carboxylate (1.00 g, 2.28 mmol, prepared
using B with 6,8-dibromo-[1,2,4]triazolo[1,5-a]pyrazine [Ark
Pharm], C with tert-butyl
3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro-1H-pyrrole-1--
carboxylate [Combi-Blocks]), MeOH (100 mL) and AcOH (4 mL). The
reaction mixture was stirred under 50 psi of H.sub.2 at rt. After 3
days, the mixture was filtered through a nylon membrane and
concentrated under reduced pressure. The crude material was
purified via silica gel chromatography eluting with 0-100%
DCM/MeOH/NH.sub.4OH (950:45:5) in DCM. The product-containing
fractions were combined, concentrated under reduced pressure, dried
under vacuum at about 70.degree. C. to give a racemic mixture of
products (0.72 g). The compound was separated via chiral prep-SFC
(Table 2, Method 2) to give (S)-tert-butyl
3-(8-((3,4-dimethoxyphenyl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)pyrr-
olidine-1-carboxylate (0.26 g, 26%) and (R)-tert-butyl
3-(8-((3,4-dimethoxyphenyl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)pyrr-
olidine-1-carboxylate (0.32 g, 32%) [Stereochemistry is arbitrarily
assigned]. LC/MS (Table 1, Method e) R.sub.t=1.45 min.; MS m/z: 441
(M+H).sup.+.
Step B.
(S)--N-(3,4-dimethoxyphenyl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1-
,5-a]pyrazin-8-amine
##STR00163##
[0423] To a solution of (S)-tert-butyl
3-(8-((3,4-dimethoxyphenyl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)pyrr-
olidine-1-carboxylate (0.25 g, 0.568 mmol, stereochemistry assigned
arbitrarily) in MeOH (2.0 mL) was added HCl (4M in 1,4-dioxane, 2.0
mL, 8.00 mmol). The reaction mixture was stirred at rt for about 4
h, then concentrated under reduced pressure. The residue was
dissolved in DCM/MeOH/NH.sub.4OH and concentrated onto silica gel
(1 g) for purification via silica gel eluting with 0-100%
DCM/MeOH/NH.sub.4OH (90:9:1) in DCM with long hold at 100%
DCM/MeOH/NH.sub.4OH (90:9:1). The product-containing fractions were
combined, concentrated, and dried under vacuum at about 70.degree.
C. to give
(S)--N-(3,4-dimethoxyphenyl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]py-
razin-8-amine (0.15 g, 78%) as an off-white solid: LC/MS (Table 1,
Method e) R.sub.t=1.45 min.; MS m/z: 341 (M+H).sup.+.
Step C.
(S)--N-(3,4-dimethoxyphenyl)-6-(1-(vinylsulfonyl)pyrrolidin-3-yl)--
[1,2,4]triazolo[1,5-a]pyrazin-8-amine
##STR00164##
[0425] To a solution of
(S)--N-(3,4-dimethoxyphenyl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-a]py-
razin-8-amine (0.073 g, 0.21 mmol) and DIEA (0.093 mL, 0.54 mmol)
in DMA (1.5 mL) at about 0.degree. C. was added ethenesulfonyl
chloride (0.025 mL, 0.22 mmol). After about 20 min, the reaction
mixture was added slowly into ice water (.about.10 mL) while
stirring. After the ice melted completely, the resulting solid was
collected via vacuum filtration and dried under vacuum at
55.degree. C. to give impure product. The solid was triturated with
MeOH (1 mL). The resulting solid was collected via vacuum
filtration while washing with additional MeOH (1 mL) and dried
under vacuum at 55.degree. C. to give solid with little change in
purity by LCMS. The filtrate and solid were recombined,
concentrated under reduced pressure and purified via silica gel
chromatography eluting with 0-5% MeOH in DCM. The
product-containing fraction was concentrated under reduced pressure
and dried under vacuum at about 55.degree. C. to give
(S)--N-(3,4-dimethoxyphenyl)-6-(1-(vinylsulfonyl)pyrrolidin-3-yl)-[1,2,4]-
triazolo[1,5-a]pyrazin-8-amine (0.024 g, 26%) as a white solid:
LC/MS (Table 1, Method e) R.sub.t=2.00 min.; MS m/z: 431
(M+H).sup.+. BTK enzyme IC.sub.50=A.
Example #4:
1-{(R)-3-[8-(6,7-Dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-ylamino)-[1,2,4]-
triazolo[1,5-a]pyridin-6-yl]-pyrrolidin-1-yl}-propenone
##STR00165##
[0426] Step A:
N-(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5-
,1-c][1,4]oxazin-2-amine
##STR00166##
[0428] A flask was charged with 1,4-dioxane (129 mL) and degassed
with nitrogen for 10 min before the addition of
8-bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine (21.5 g, 64.7 mmol,
Example #1, Step A),
6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-amine (9.91 g, 71.2
mmol, Preparation #1),
(9,9-dimethyl-9H-xanthene-4,5-diyl)bis(diphenylphosphine) (6.74 g,
11.65 mmol), diacetoxypalladium (1.308 g, 5.83 mmol), and cesium
carbonate (63.3 g, 194 mmol). The reaction mixture was degassed
with nitrogen for about 10 min., and then heated to about
120.degree. C. for about 1 h. The reaction cooled to ambient
temperature and 400 mL of water was slowly added and stirred
vigorously. The resulting precipitate was filtered and dried in the
vacuum oven overnight to afford
N-(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5-
,1-c][1,4]oxazin-2-amine (23.8 g, 102%, 85% purity): LC/MS (Table
1, Method 1) R.sub.t=1.79 min.; MS m/z: 292, 294 (M+H).sup.+.
Step B: tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate
##STR00167##
[0430] An oven-dried 2 L 3-necked flask was charged with
N-(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5-
,1-c][1,4]oxazin-2-amine (23 g, 79 mmol), tert-butyl
3-(4,4,5,5-tetramethyl-1,3-dioxolan-2-yl)-2,5-dihydro-1H-pyrrole-1-carbox-
ylate (40.0 g, 134 mmol, Combi Blocks), 1,4-dioxane (330 mL) and a
solution of potassium phosphate (40.3 g, 190 mmol) in water (66.0
mL). The reaction was sparged with argon for 25 min, followed by
the addition of Xphos palladacycle G1 (2.92 g, 3.96 mmol). The
reaction mixture was sparged with argon for about 30 min then
heated to about 60.degree. C. for about 80 min. The reaction cooled
to ambient temperature and about 400 mL of water was slowly added
to the reaction mixture. The resulting precipitate was collected
via filtration and dried in the vacuum oven. The filtercake was
taken up in DCM (1L) and washed with brine (350 mL), dried over
anhydrous MgSO.sub.4, filtered, and concentrated under reduced
pressure. The solid was then triturated with about 200 mL of
ditheylether. The resulting precipitate was collected via
filtration and dried in the vac oven to afford tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (31.3
g, 93%): LC/MS (Table 1, Method 1) R.sub.t=2.10 min.; MS m/z: 424
(M+H).sup.+.
Step C: (R)-tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)pyrrolidine-1-carboxylate and (S)-tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)pyrrolidine-1-carboxylate
##STR00168##
[0432] EtOH (0.50 mL) was added to tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (5.24
mg, 0.012 mmol) and 20% wt. palladium hydroxide on carbon (1.06 mg,
7.55 .mu.mol) in a 4 mL pressure bottle. The mixture was stirred
under 60 psi of hydrogen at about 50.degree. C. for about 2 h. The
catalyst was filtered off through a pad of Celite.RTM. and the
remaining solvent was concentrated under reduced pressure. The
crude reaction mixture was purified by silicagel chromatography
eluting with 10-60% EtOAc/DCM to afford tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)pyrrolidine-1-carboxylate (14.5 g, 47%). The
racemic mixture was then purified via chiral preparative HPLC
(Table 2, Method 9) to afford (R)-tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)pyrrolidine-1-carboxylate (3.79 g, 12%,
OR=+) and (S)-tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)pyrrolidine-1-carboxylate (4.67, 15%, OR=-):
LC/MS (Table 1, Method 1) R.sub.t=2.03 min.; MS m/z: 426
(M+H).sup.+.
Step D:
(R)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino-
)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one
##STR00169##
[0434] A flask was charged with (R)-tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)pyrrolidine-1-carboxylate (3.7 g, 8.70 mmol)
and 1,4-dioxane (43 mL) with hydrogen chloride (4 N in 1,4-dioxane,
21 mL, 87 mmol). The reaction was heated to about 35.degree. C. for
about 90 min. The solvent was concentrated under reduced pressure.
The remaining residue was taken up in DCM (43 mL) and cooled to
about -40.degree. C., followed by the addition of
N-ethyl-N-isopropylpropan-2-amine (7.59 mL, 43.5 mmol) and acryloyl
chloride (0.918 mL, 11.30 mmol). The reaction stirred at about
-40.degree. C. for about 10 min. The solvent was concentrated under
reduced pressure. The crude residue was purified by silica gel
chromatography eluting with 0-8% MeOH/DCM. The compound was then
recrystallized from MeCN using 10:1 volume ratio to afford
(R)-1-(3-(8-((6,7-dihydro-4H-pyrazolo
[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)pyrroli-
din-1-yl)prop-2-en-1-one (2.4 g, 72.7% OR=+): LC/MS (Table 1,
Method 1) R.sub.t=1.41 min.; MS m/z: 380 (M+H).sup.+. BTK enzyme
IC.sub.50=A
Example #5.
(R)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,-
4]triazolo[1,5-a]pyridin-6-yl)piperidin-1-yl)prop-2-en-1-one
##STR00170##
[0435] Step A.
N-(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5-
,1-c][1,4]oxazin-2-amine
##STR00171##
[0437] 8-Bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine (0.500 g,
2.151 mmol, Example #1, Step A),
6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-amine (0.329 g, 2.366
mmol, Preparation #1), Xantphos (0.261 g, 0.452 mmol),
Pd(OAc).sub.2 (0.048 g, 0.215 mmol) and Cs.sub.2CO.sub.3 (1.752 g,
5.38 mmol) in 1,4-dioxane (5 mL) was sparged with nitrogen. The
solution was then heated in a microwave at about 120.degree. C. for
about 1.5 h. The reaction mixture was cooled to rt, partitioned
between DCM (3.times.20 mL) and water (20 mL). The organic layers
were combined, concentrated, purified via silica gel eluting with
20-60% EtOAc in heptanes to give
N-(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5-
,1-c][1,4]oxazin-2-amine (0.348 g, 55.7%): LC/MS (Table 1, Method
1) R.sub.t=1.82 min.; MS m/z: 291 (M+H).sup.+.
Step B. tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)-5,6-dihydropyridine-1(2H)-carboxylate
##STR00172##
[0439] To a 10 mL microwave tube were added
N-(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5-
,1-c][1,4]oxazin-2-amine (0.576 g, 1.981 mmol), tert-butyl
3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-
-carboxylate (0.858 g, 2.77 mmol, Anichem), THF (3 mL), MeOH (0.600
mL) and 2 M aqueous sodium carbonate solution (2.97 mL, 5.94 mmol).
The mixture was sparged with nitrogen, followed by addition of
1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloride
dichloromethane complex (0.162 g, 0.198 mmol). The vial was sealed
and sparged with nitrogen again. The reaction mixture was heated in
microwave at about 130.degree. C. for about 1 h. The reaction
mixture was filtered through Celite.RTM., washed with DCM and MeOH,
concentrated under reduced pressure. The resulting residue was
purified via silica gel chromatography eluting with 0-10% DCM/MeOH
to give tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)-5,6-dihydropyridine-1(2H)-carboxylate
(0.704 g, 65.0%) as a light brown solid: LC/MS (Table 1, Method 1)
R.sub.t=2.19 min.; MS m/z: 438 (M+H).sup.+.
Step C. (S)-tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)piperidine-1-carboxylate and (R)-tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)piperidine-1-carboxylate
##STR00173##
[0441] A solution of tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)-5,6-dihydropyridine-1(2H)-carboxylate
(0.704 g, 1.609 mmol) in THF (15 mL), MeOH (15 mL) and AcOH (10 mL)
was passed through H-cube with Pearlman's catalyst Catcart.RTM. at
about 45.degree. C. under about 50 bar of H.sub.2 for about 16 h.
LC/MS showed complete conversion. The solvent was concentrated
under reduced pressure. The racemic mixture was then separated via
chiral prep HPLC (Table 2, Method 7) to give (S)-tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)piperidine-1-carboxylate (0.050 g, 7.07%)
and (R)-tert-butyl 3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][,
4]oxazin-2-yl)amino)-[1,2,4]triazolo[,
5-a]pyridin-6-yl)piperidine-1-carboxylate (0.053 g, 7.49%)
[Stereochemistry is arbitrarily assigned]. LC/MS (Table 1, Method
1) R.sub.t=2.17 min.; MS m/z: 440 (M+H).sup.+.
Step D.
(R)--N-(6-(piperidin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-
-dihydro-4H-pyrazolo [5,1-c][1,4]oxazin-2-amine
##STR00174##
[0443] A flask was charged with MeOH (3.0 mL) and cooled to about
0.degree. C. Acetyl chloride (0.364 mL, 5.12 mmol) was added
dropwise. The mixture was stirred at rt for about 2 h. The solution
was then added to (R)-tert-butyl
3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]triaz-
olo[1,5-a]pyridin-6-yl)piperidine-1-carboxylate (0.050 g, 0.114
mmol). The reaction mixture was stirred at rt over night. The
reaction mixture was concentrated to give
(R)--N-(6-(piperidin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydr-
o-4H-pyrazolo[5,1-c][1,4]oxazin-2-amine as a light yellow solid.
LC/MS (Table 1, Method 1) R.sub.t=1.12 min; MS m/z: 340
(M+H).sup.+. The crude material was used in the next step without
further purification.
Step E.
(R)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino-
)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)piperidin-1-yl)prop-2-en-1-one
##STR00175##
[0445] A flask was charged with
(R)--N-(6-(piperidin-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydr-
o-4H-pyrazolo[5,1-c][1,4]oxazin-2-amine (0.039 g, 0.115 mmol) and
TEA (0.072 mL, 0.517 mmol) in THF (1.1 mL). The reaction mixture
was cooled to about 0.degree. C. in an ice-bath. Acryloyl chloride
(0.01 mL, 0.126 mmol) was added. The mixture was stirred for about
20 min, then was diluted with water (1.0 mL). The reaction mixture
was purified by prep-HPLC (Table 1, Method k) to afford
(R)-1-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,-
4]triazolo[1,5-a]pyridin-6-yl)piperidin-1-yl)prop-2-en-1-one (0.016
g, 35%, OR=+) as an off-white solid: LC/MS (Table 1, Method h)
R.sub.t=1.58 min; MS m/z: 394 (M+H).sup.+. BTK enzyme
IC.sub.50=A.
Example #6.
(S)-1-(3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyraz-
in-6-yl)pyrrolidin-1-yl)prop-2-en-1-one
##STR00176##
[0446] Step A.
6-bromo-N-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amin-
e
##STR00177##
[0448] To a mixture of NaH (0.864 g, 60%, 36 mmol) in THF (50 mL)
was added 1-methyl-1H-pyrazol-4-amine (2.097 g, 21.59 mmol). The
mixture was stirred for about 0.5 h at about 0.degree. C. A
solution of 6,8-dibromo-[1,2,4]triazolo[1,5-a]pyrazine (5 g, 17.99
mmol, Ark Pharm) in THF (30 mL) was added slowly and the reaction
mixture was stirred for 1 h at 0.degree. C. Water (5 mL) was added.
The mixture was concentrated to dryness. Additional two reaction
mixtures were set up following the above method. All three batches
were combined, water (100 mL) was added. The mixture was extracted
with EtOAc (3.times.500 mL). The organic phase was dried over
Na.sub.2SO.sub.4, concentrated under reduced pressure, purified via
silica gel chromatography eluting with 25-50% EtOAc/PE to give
6-bromo-N-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-
-amine (12 g, 76%) as a white solid: .sup.1H NMR (400 MHz, DMSO-D6)
.delta. ppm .delta. 10.71 (s, 1H), 8.59-8.56 (d, J=12, 2H), 8.07
(s, 1H), 7.76 (s, 1H), 3.86 (s, 3H).
Step B. tert-butyl
3-(8-(1-methyl-1H-pyrazol-4-ylamino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)--
2,5-dihydro-1H-pyrrole-1-carboxylate
##STR00178##
[0450] To a solution of
6-bromo-N-(l-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amin-
e (5 g, 17 mmol) in 1.4-dioxane (90 mL) and water (30 mL) was added
tert-butyl
3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro-1H-pyrrole-1--
carboxylate (10.04 g, 34 mmol, Combi-Blocks), Cs.sub.2CO.sub.3
(16.62 g, 51 mmol) and Pd(PPh.sub.3).sub.2Cl.sub.2 (0.747 g, 1.7
mmol). The mixture was stirred for about 12 h at about 120.degree.
C. under N.sub.2 protection. The reaction mixture was concentrated
under reduced pressure. The residue was purified via silica gel
chromatography eluting with 25-50% EtOAc in PE to give tert-butyl
3-(8-(1-methyl-1H-pyrazol-4-ylamino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)--
2,5-dihydro-1H-pyrrole-1-carboxylate (4.5 g, 69%) as a white solid:
.sup.1H NMR (400 MHz, DMSO-D6) .delta. ppm 10.31-10.30 (d, J=4,
1H), 8.55 (s, 1H), 8.37-8.30 (d, J=28, 1H), 8.13-8.10 (d, J=12,
1H), 7.79-7.75 (d, J=16, 1H), 6.67-6.64 (d, J=12, 1H), 4.46-4.43
(d, J=12, 2H), 4.25 (s, 2H), 3.30 (s, 3H), 1.46-1.44 (d, J=8,
9H).
Step C. tert-butyl
3-(8-(1-methyl-1H-pyrazol-4-ylamino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)p-
yrrolidine-1-carboxylate
##STR00179##
[0452] To a solution of tert-butyl
3-(8-(1-methyl-1H-pyrazol-4-ylamino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)--
2,5-dihydro-1H-pyrrole-1-carboxylate (4.5 g, 11.77 mmol) in THF
(500 mL) was added Pd/C (4.5 g, 10%, 4.23 mmol) under protection of
argon. The reaction mixture was stirred for about 12 h at room
temperature under H.sub.2 (about 14 psi). The reaction mixture was
filtered through a pad of Celite.RTM., and the filtrate was
concentrated under reduced pressure to give tert-butyl
3-(8-(1-methyl-H-pyrazol-4-ylamino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)py-
rrolidine-1-carboxylate (4.16 g, 92%) as a white solid: LC/MS
(Table 1, Method d) R.sub.t=2.94 min; MS m/z: 385 (M+H).sup.+.
Step D. (S)-tert-butyl
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)pyrrolidine-1-carboxylate and (R)-tert-butyl
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)pyrrolidine-1-carboxylate
##STR00180##
[0454] tert-Butyl
3-(8-(1-methyl-1H-pyrazol-4-ylamino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)p-
yrrolidine-1-carboxylate was separated via chiral prep HPLC (Table
2, Method 6) to give (S)-tert-butyl
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)pyrrolidine-1-carboxylate (0.435 g, 40.2%, OR=+) and
(R)-tert-butyl
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[,1,5-a]pyrazin-6-y-
l)pyrrolidine-1-carboxylate (0.442 g, 40.9%, OR=-) both as white
solids. [Stereochemistry is arbitrarily assigned]. LC/MS (Table 1,
Method 1) R.sub.t=1.96 min.; MS m/z: 385 (M+H).sup.+.
Step E.
(S)--N-(1-methyl-1H-pyrazol-4-yl)-6-(pyrrolidin-3-yl)-[1,2,4]triaz-
olo[1,5-a]pyrazin-8-amine
##STR00181##
[0456] A flask was charged with MeOH (30 mL) and cooled to about
0.degree. C. Acetyl chloride (3.60 mL, 50.9 mmol) was added
dropwise. The mixture was stirred at rt for about 2 h. The solution
was then added to (S)-tert-butyl
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)pyrrolidine-1-carboxylate (0.435 g, 1.132 mmol). The reaction
mixture was stirred at rt for about 3 h. Solvent was removed to
give
(S)--N-(1-methyl-1H-pyrazol-4-yl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-
-a]pyrazin-8-amine as an off-white solid. LC/MS (Table 1, Method 1)
R.sub.t=0.99 min; MS m/z: 285 (M+H).sup.+. The crude material was
used in the next without further purification.
Step F.
(S)-1-(3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5--
a]pyrazin-6-yl)pyrrolidin-1-yl)prop-2-en-1-one
##STR00182##
[0458] To a solution of
(S)--N-(1-methyl-1H-pyrazol-4-yl)-6-(pyrrolidin-3-yl)-[1,2,4]triazolo[1,5-
-a]pyrazin-8-amine (0.200 g, 0.703 mmol), TEA (0.49 mL, 3.52 mmol)
in DMF (3.5 mL) was added acryloyl chloride (0.057 mL, 0.703 mmol).
The reaction mixture was stirred at rt for about 30 min. Water (1.5
mL) was added to quench the reaction. The mixture was purification
by prep-HPLC (Table 1, Method k) to give
(S)-1-(3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyraz-
in-6-yl)pyrrolidin-1-yl)prop-2-en-1-one (0.075 g, 31%, OR=+) as a
white solid: LC/MS (Table 1, Method h) R.sub.t=1.42 min; MS m/z:
339 (M+H).sup.+. BTK enzyme IC.sub.50=A.
Example #7:
N-((1R,3S)-3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclopentyl)acrylamide
##STR00183##
[0459] Step A:
6-Chloro-N-(1-methyl-1H-pyrazol-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-ami-
ne
##STR00184##
[0461] Cesium carbonate (42.0 g, 129 mmol) was added to 1,4-dioxane
(238 mL) to give a white suspension. The mixture was degassed with
nitrogen, then 8-bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine (10
g, 43.0 mmol) (Example #1 Step A),
(9,9-dimethyl-9H-xanthene-4,5-diyl)bis(diphenylphosphine) (4.98 g,
8.60 mmol) and 1-methyl-1H-pyrazol-3-amine (3.66 mL, 43.0 mmol)
(Combi-Blocks) were each added sequentially rapidly to the mixture.
The mixture was degassed with nitrogen, diacetoxypalladium (0.966
g, 4.30 mmol) was added. The mixture was further degassed with
nitrogen and heated at about 120.degree. C. for about 1 h. The
mixture was allowed to cool to room temperature and EtOAc (250 mL)
was added. The mixture was stirred and filtered through
Celite.RTM., washed with EtOAc (5.times.50 mL), and concentrated to
give black syrup, which was partitioned between DCM (250 mL) and
water (100 mL). The organic layer was drained, the aqueous layer
was extracted by DCM (100 mL), the combined DCM layers were dried
over sodium sulfate, filtered and concentrated to afford black
syrup, which was deposited on silica gel (75 g), purified by silica
gel chromatography eluting with a gradient of 20-55% EtOAc/heptane
to afford
6-chloro-N-(1-methyl-1H-pyrazol-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-ami-
ne (6.9 g, 64.5%): LC/MS (Table 1, Method a) R.sub.t=1.81 min; MS
m/z 249 (M+H).sup.+.
Step B: tert-butyl
(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(1-methyl-1H-pyrazol-3-yl)ca-
rbamate
##STR00185##
[0463]
6-Chloro-N-(1-methyl-1H-pyrazol-3-yl)-[1,2,4]triazolo[1,5-a]pyridin-
-8-amine (5 g, 20.11 mmol) and di-tert-butyl dicarbonate (8.78 g,
40.2 mmol) in DCM (101 mL) were combined to give a pale yellow
solution. N,N-Dimethylpyridin-4-amine (0.123 g, 1.005 mmol) was
added and the reaction mixture was stirred at rt for about 18 h.
The mixture was concentrated to give a yellow solid which was mixed
with EtOAc (200 mL) and 2-MeTHF (50 mL), washed with citric acid
(10% in water, 2.times.75 mL), saturated aqueous sodium bicarbonate
(4.times.60 mL) and brine (60 mL). To the organic layer was added
DCM (100 mL), and the solution was dried over magnesium sulfate
(13.7 g), filtered and concentrated to afford tert-butyl
(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(1-methyl-1H-pyrazol-3-yl)ca-
rbamate (7.11 g, 100%): LC/MS (Table 1, Method a) R.sub.t=1.94 min;
MS m/z 349 (M+H).sup.+.
Step C: tert-butyl
(1-methyl-1H-pyrazol-3-yl)(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl-
)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)carbamate
##STR00186##
[0465] Potassium acetate (5.99 g, 61.1 mmol) and tert-butyl
(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(1-methyl-1H-pyrazol-3-yl)ca-
rbamate (7.1 g, 20.36 mmol) was added in 1,4-dioxane (70.9 mL) to
give an orange suspension. The mixture was degassed with nitrogen
then bis(pinacolato)diboron (12.92 g, 50.9 mmol) and XPhos (0.776
g, 1.628 mmol) were added sequentially rapidly to the mixture. The
mixture was degassed with nitrogen, Pd.sub.2(dba).sub.3 (0.373 g,
0.407 mmol) was added, the mixture was degassed with nitrogen, then
it was heated at about 110.degree. C. for about 18 h. The reaction
mixture was allowed to cool to rt and DCM (30 mL) was added and
stirred, the mixture was diluted with EtOAc (100 mL), filtered
through Celite.RTM., and washed by EtOAc (5.times.30 mL), and
concentrated to afford a red solid. The solid was dissolved in DCM
(120 mL) and heptane (120 mL), the red solution was concentrated
under reduced pressure to remove most DCM and some heptane. The
resulting suspension was filtered, washed with heptane (3.times.10
mL), dried in a vacuum oven to afford tert-butyl
(1-methyl-1H-pyrazol-3-yl)(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl-
)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)carbamate (8.765 g, 98%):
LC/MS (Table 1, Method a) R.sub.t=1.42 min; MS m/z 441
(M+H).sup.+.
Step D: tert-butyl
(1-methyl-1H-pyrazol-3-yl)(6-(3-oxocyclopent-1-en-1-yl)-[1,2,4]triazolo[1-
,5-a]pyridin-8-yl)carbamate
##STR00187##
[0467] tert-Butyl
(1-methyl-1H-pyrazol-3-yl)(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl-
)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)carbamate (3.8 g, 8.63 mmol)
was added to 1,4-dioxane (28.8 mL) to give cloudy yellow solution.
Potassium phosphate (5.50 g, 25.9 mmol) in water (9.59 mL) was
added to the solution. The reaction mixture was degassed with
nitrogen, XPhos palladacycle (0.319 g, 0.432 mmol) was added and
degassed with nitrogen; then a solution of 3-bromocyclopent-2-enone
(2.62 mL, 23.91 mmol) (SynTech) in 1,4-dioxane (3.times.6 mL) were
added rapidly., The mixture was degassed with nitrogen, then heated
at about 60.degree. C. for about 18 h. The reaction mixture was
cooled to room temperature and diluted with DCM (100 mL), filtered
through Celite.RTM., washed with DCM (5.times.30 mL), concentrated
to give a brown syrup, which was deposited on silica gel (20 g),
purified by silica gel chromatography eluting with a gradient of 2
to 5% MeOH/DCM to afford tert-butyl
(1-methyl-1H-pyrazol-3-yl)(6-(3-oxocyclopent-1-en-1-yl)-[1,2,4]triazolo[1-
,5-a]pyridin-8-yl)carbamate (2.54 g, 73%): LC/MS (Table 1, Method
a) R.sub.t=1.63 min; MS m/z 395 (M+H).sup.+.
Step E: (S)-tert-butyl
(1-methyl-1H-pyrazol-3-yl)(6-(3-oxocyclopentyl)-[1,2,4]triazolo[1,5-a]pyr-
idin-8-yl)carbamate
##STR00188##
[0469]
(2R,5R)-5-Benzyl-3-methyl-2-(5-methylfuran-2-yl)imidazolidin-4-one
(0.014 g, 0.051 mmol) and tert-butyl
(1-methyl-1H-pyrazol-3-yl)(6-(3-oxocyclopent-1-en-1-yl)-[1,2,4]triazolo[1-
,5-a]pyridin-8-yl)carbamate (0.1 g, 0.254 mmol) were added to THF
(0.507 mL) to give a yellow suspension. The mixture was cooled to
about 0.degree. C. in an ice bath. Diethyl
2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylate (0.077 g, 0.304
mmol) and trichloroacetic acid (5.11 .mu.L, 0.051 mmol) were added.
The mixture was stirred for about 44 h, the cooling bath was warmed
up by ambient air. Then the crude was purified by silica gel
chromatography eluting with a gradient of 0 to 5% MeOH/DCM to
afford (S)-tert-butyl
(1-methyl-1H-pyrazol-3-yl)(6-(3-oxocyclopentyl)-[1,2,4]triazolo[1,5-a]pyr-
idin-8-yl)carbamate (0.088 g, 88%, OR=negative) [Stereochemistry is
arbitrarily assigned]: LC/MS (Table 1, Method a) R.sub.t=1.65 min;
MS m/z 397 (M+H).sup.+.
Step F: tert-butyl
(6-((1S,3R)-3-((R)-1,1-dimethylethylsulfinamido)cyclopentyl)-[1,2,4]triaz-
olo[1,5-a]pyridin-8-yl)(1-methyl-1H-pyrazol-3-yl)carbamate
##STR00189##
[0471] (S)-tert-butyl
(1-methyl-1H-pyrazol-3-yl)(6-(3-oxocyclopentyl)-[1,2,4]triazolo[1,5-a]pyr-
idin-8-yl)carbamate (14 g, 35.3 mmol) and
(R)-2-methylpropane-2-sulfinamide (6.42 g, 53.0 mmol) were added in
THF (72.7 mL) to give an orange solution. The mixture was degassed
with nitrogen, then tetraethoxytitanium (24.17 g, 106 mmol) was
added, the solution was heated at about 50.degree. C. for about 18
h. The reaction mixture was cooled to rt, then the solution was
cooled to about -50.degree. C. in a dry ice/MeCN bath, sodium
borohydride (1.924 g, 50.9 mmol) was added in one portion; the
reaction mixture was stirred and the cooling bath was warmed up
gradually over 4 h period. The resulting red solution was added
dropwise into stirring aqueous sodium chloride solution (24%, 400
mL). THF (100 mL) and 2-MeTHF (200 mL) were added and the solution
was stirred for about 1 h, The top organic layer was decanted by
suction, the aqueous suspension was added 2-MeTHF (200 mL) and
stirred for about 30 min, then it was filtered through Celite.RTM.,
washed by 2-MeTHF (4.times.50 mL), the filtrate was partitioned,
the organic layers were combined, washed by saturated aqueous
sodium bicarbonate (2.times.100 mL) and brine (100 mL), dried over
magnesium sulfate, filtered and concentrated to afford 20 g yellow
solid, which was purified by silica gel chromatography eluting with
a gradient of 0 to 6.5% MeOH/DCM to afford 14.6 g yellow solid. The
mixture was separated via chiral prep (Table 2, Method 8) to give
tert-butyl
(6-((1S,3R)-3-((R)-1,1-dimethylethylsulfinamido)cyclopentyl)-[1,2,4]triaz-
olo[1,5-a]pyridin-8-yl)(1-methyl-1H-pyrazol-3-yl)carbamate (7.0 g,
39.5%, OR=negative) [Stereochemistry is arbitrarily assigned]:
LC/MS (Table 1, Method a) R.sub.t=1.90 min; MS m/z 502
(M+H).sup.+.
Step G:
6-((1S,3R)-3-aminocyclopentyl)-N-(1-methyl-1H-pyrazol-3-yl)-[1,2,4-
]triazolo[1,5-a]pyridin-8-amine hydrochloride
##STR00190##
[0473] tert-Butyl
(6-((1S,3R)-3-((R)-1,1-dimethylethylsulfinamido)cyclopentyl)-[1,2,4]triaz-
olo[1,5-a]pyridin-8-yl)(1-methyl-1H-pyrazol-3-yl)carbamate (5.5 g,
10.96 mmol) was added in MeOH (22.02 mL) to give a pale yellow
solution. The solution was cooled to about 0.degree. C. in an ice
bath. Hydrochloric acid (3.0 M in cyclopentyl methyl ether) (43.9
mL, 132 mmol) was added dropwise via addition funnel. The ice bath
was removed after stirring for about 1 h, then the mixture was
stirred at rt for about 18 h. Ether (100 mL) was added and the
solution was stirred for 1 h. To the suspension was added ether (50
mL) and the mixture was filtered. The collected solid was rinsed
with ether (5.times.20 mL), dried over 1 h to afford
6-((1S,3R)-3-aminocyclopentyl)-N-(1-methyl-1H-pyrazol-3-yl)-[1,2,4]triazo-
lo[1,5-a]pyridin-8-amine hydrochloride: LC/MS (Table 1, Method a)
R.sub.t=1.04 min; MS m/z 298 (M+H).sup.+. It was used as is in next
step.
Step H:
N-((1R,3S)-3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[-
1,5-a]pyridin-6-yl)cyclopentyl)acrylamide
##STR00191##
[0475]
6-((1S,3R)-3-Aminocyclopentyl)-N-(1-methyl-1H-pyrazol-3-yl)-[1,2,4]-
triazolo[1,5-a]pyridin-8-amine hydrochloride (3.66 g, 10.96 mmol)
was added in 2-MeTHF (60 mL) to give a white suspension. The
mixture was cooled to about 0.degree. C. in an ice bath. A solution
of potassium hydrogenphosphate (22.91 g, 132 mmol) in water (70 mL)
was added dropwise via dropping funnel and the mixture was stirred
for about 10 min. A solution of acryloyl chloride (0.890 mL, 10.96
mmol) in 2-MeTHF (10 mL) was added dropwise via syringe over about
15 min, the reaction mixture was stirred at about 0.degree. C. for
about 30 min. The mixture was partitioned, the aqueous layer was
drained, the organic layer was washed by saturated aqueous sodium
bicarbonate (2.times.50 mL), brine (50 mL), dried over sodium
sulfate, filtered and concentrated to afford 3.63 g pale yellow
solid. MeCN (40 mL) was added and the suspension was stirred for
about 1 h, then it was filtered, the collected solid was rinsed by
ice-cold MeCN (4.times.10 mL), pentane (7.times.20 mL) to afford
N-((1R,3S)-3-(8-((1-methyl-1H-pyrazol-3-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclopentyl)acrylamide (2.5 g, 64.9% OR=positive)
[Stereochemistry is arbitrarily assigned]: LC/MS (Table 1, Method
a) R.sub.t=1.48 min; MS m/z 352 (M+H).sup.+. BTK enzyme
IC.sub.50=A
Example #8:
N-((1R,3S)-3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[-
1,2,4]triazolo[1,5-a]pyridin-6-yl)cyclopentyl)acrylamide and
N-((1S,3R)-3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[-
1,2,4]triazolo[1,5-a]pyridin-6-yl)cyclopentyl)acrylamide
##STR00192##
[0476] Step A:
N-(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo
[5,1-c][1,4]oxazin-2-amine
##STR00193##
[0478] To a microwave reaction vial were added tert-butyl
8-bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine (1.0 g, 4.3 mmol,
Example #1, Step A),
6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-amine (0.658 g, 4.73
mmol, Preparation #1), 1,4-dioxane (10 mL), Cs.sub.2CO.sub.3 (2.80
g, 8.60 mmol), Xantphos (0.124 g, 0.215 mmol) and
Pd.sub.2(dba).sub.3 (0.197 g, 0.215 mmol), The reaction vial was
flushed with nitrogen, capped, stirred and heated to about
120.degree. C. in a Biotage microwave reactor for about 3 h. The
reaction mixture was diluted with DCM (80 mL) and water (50 mL).
The organic layer was separated, washed with water (50 mL), brine
(50 mL), and dried over Na.sub.2SO.sub.4. The organic layer was
filtered, concentrated under reduced pressure and purified via
silica gel chromatography eluting with 5% MeOH in DCM to afford
N-(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5-
,1-c][1,4]oxazin-2-amine (0.85 g, 61.2%) as a yellow solid: LC/MS
(Table 1, Method m) R.sub.t=1.56 min; MS m/z 291 (M+H).sup.+.
Step B: tert-butyl
(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-dihydro-4H-pyrazolo
[5,1-c][1,4]oxazin-2-yl)carbamate
##STR00194##
[0480] A mixture of
N-(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5-
,1-c][1,4]oxazin-2-amine (2.0 g, 6.9 mmol), BoC.sub.2O (4.79 mL,
20.6 mmol), TEA (2.88 mL, 20.6 mmol) and DMAP (0.840 g, 6.88 mmol)
in DCM (100 mL) was stirred at rt overnight. The organic layer was
washed with saturated NH.sub.4Cl (3.times.50 mL). The organic layer
was dried with Na.sub.2SO.sub.4, filtered and concentrated under
reduced pressure. The crude product was recrystallized from
petroleum ether (60.about.90.degree. C.) to give tert-butyl
(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-dihydro-4H-pyrazolo[5,1-
-c][1,4]oxazin-2-yl)carbamate (2.5 g, 86%) as a yellow solid: LC/MS
(Table 1, Method n) R.sub.t=1.72 min; MS m/z 391(M+H).sup.+.
Step C: tert-butyl
(6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)(6-(3-oxocyclopent-1-en-1-
-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)carbamate
##STR00195##
[0482] A round bottom flask was charged with tert-butyl
(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-dihydro-4H-pyrazolo[5,1-
-c][1,4]oxazin-2-yl)carbamate (0.05 g, 0.13 mmol),
dicyclohexyl(2',6'-dimethoxy-[1,1'-biphenyl]-2-yl)phosphine (0.016
g, 0.038 mmol), potassium phosphate (0.081 g, 0.38 mmol), water
(0.4 mL) and toluene (4 mL). The reaction mixture was degassed with
nitrogen followed by the addition of
3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopent-2-enone
(0.035 g, 0.17 mmol, US20120077814) and Pd.sub.2(dba).sub.3 (0.012
g, 0.013 mmol). The suspension was heated in a Biotage microwave at
about 100.degree. C. for about 2 h. The mixture was purified by
column chromatography (DCM: MeOH=40:1) to give tert-butyl
(6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)(6-(3-oxocyclopent-1-en-1-
-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)carbamate (0.03 g, 53.7%):
LC/MS (Table 1, Method r) R.sub.t=1.57 min; MS m/z
437(M+H).sup.+.
Step D: tert-butyl
(6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)(6-(3-oxocyclopentyl)-[1,-
2,4]triazolo[1,5-a]pyridin-8-yl)carbamate
##STR00196##
[0484] A round bottom flask was charged with tert-butyl
(6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)(6-(3-oxocyclopent-1-en-1-
-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)carbamate (0.04 g, 0.09
mmol) in MeOH (15 mL) followed by the addition of 10% Pd/C (0.010
g, 0.092 mmol). The suspension was stirred under an atmosphere of
hydrogen at rt for 1 day. The suspension was filtered, and the
filtrate was concentrated under reduced pressure to give tert-butyl
(6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)(6-(3-oxocyclopentyl)-[1,-
2,4]triazolo[1,5-a]pyridin-8-yl)carbamate (0.03 g 75%): LC/MS
(Table 1, Method r) R.sub.t=1.57 min; MS m/z 439 (M+H).sup.+.
Step E: tert-butyl
(6-(3-(benzylamino)cyclopentyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-d-
ihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)carbamate
##STR00197##
[0486] A solution of tert-butyl
(6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)(6-(3-oxocyclopentyl)-[1,-
2,4]triazolo[1,5-a]pyridin-8-yl)carbamate (0.080 g, 0.18 mmol) in
DCM (10 mL) was treated with AcOH (0.01 mL, 0.182 mmol) followed by
phenylmethanamine (0.098 g, 0.91 mmol). After stirring for about 20
min at rt under nitrogen, sodium triacetoxyborohydride (0.193 g,
0.912 mmol) was added and stirring was continued overnight. MeOH (2
mL), DCM (10 mL) and saturated NaCl (10 mL) were added and the
layers were separated. The aqueous layer was extracted with DCM (10
mL). The product was purified by Prep-TLC to give tert-butyl
(6-(3-(benzylamino)cyclopentyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-d-
ihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)carbamate (0.03 g,
27.9%): LC/MS (Table 1, Method m) R.sub.t=1.81, 1.83 min; MS m/z
530 (M+H).sup.+.
Step F: tert-butyl
(6-(3-aminocyclopentyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-dihydro-4-
H-pyrazolo[5,1-c][1,4]oxazin-2-yl)carbamate
##STR00198##
[0488] A mixture of tert-butyl
(6-(3-(benzylamino)cyclopentyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-d-
ihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)carbamate (0.04 0 g,
0.076 mmol), 10% Pd/C (0.10 g, 0.944 mmol) and ammonium formate
(0.30 g, 4.8 mmol) in MeOH (20 mL) were refluxed under nitrogen for
about 2 h. The reaction mixture was cooled, filtered through
Celite.RTM., and concentrated. The residue was diluted with
saturated NaCl (30 mL) and extracted with DCM (3.times.30 mL). The
combined organic layers were washed with water (50 mL) and brine
(50 mL), then were dried over anhydrous sodium sulfate, filtered
and concentrated to give crude tert-butyl
(6-(3-aminocyclopentyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-dihydro-4-
H-pyrazolo[5,1-c][1,4]oxazin-2-yl)carbamate (0.025 g, 52.7%): LC/MS
(Table 1, Method m) R.sub.t=1.39 min; MS m/z 440 (M+H).sup.+.
Step G:
N-(6-(3-aminocyclopentyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7--
dihydro-4H-pyrazolo [5,1-c][1,4]oxazin-2-amine
##STR00199##
[0490] A round bottom flask was charged with tert-butyl
(6-(3-aminocyclopentyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)(6,7-dihydro-4-
H-pyrazolo[5,1-c][1,4]oxazin-2-yl)carbamate (0.120 g, 0.273 mmol)
and MeOH (6 mL). A solution of 4M HCl in dioxane (0.010 mL, 0.28
mmol) was added and the reaction was stirred at rt for about 4 h.
The solution was concentrated to dryness to give
N-(6-(3-aminocyclopentyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-
-4H-pyrazolo[5,1-c][1,4]oxazin-2-amine (0.085 g, 60%): LC/MS (Table
1, Method m) R.sub.t=1.24 min; MS m/z 340 (M+H).sup.+.
Step H:
N-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1-
,2,4]triazolo[1,5-a]pyridin-6-yl)cyclopentyl)acrylamide
##STR00200##
[0492] A round bottom flask was charged with
N-(6-(3-aminocyclopentyl)-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-
-4H-pyrazolo[5,1-c][1,4]oxazin-2-amine hydrochloride (0.102 g,
0.271 mmol) and DCM (6 mL) and the solution was cooled to about
0.degree. C. To the flask was added TEA (0.378 mL, 2.71 mmol) and
the solution was stirred for about 10 min followed by the dropwise
addition of a solution of acryloyl chloride (0.032 g, 0.353 mmol)
in DCM (0.1 mL). The mixture was stirred for about 20 min. The
reaction solution was then concentrated under reduced pressure to
give
N-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[1,2,4]tr-
iazolo[1,5-a]pyridin-6-yl)cyclopentyl)acrylamide (0.05 g, 46.8%):
LC/MS (Table 1, Method m) R.sub.t=1.42 min; MS m/z 394
(M+H).sup.+.
Step I:
N-((1R,3S)-3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)a-
mino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)cyclopentyl)acrylamide and
N-((1S,3R)-3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[-
1,2,4]triazolo[1,5-a]pyridin-6-yl)cyclopentyl)acrylamide
##STR00201##
[0494] The
N-(3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-
-[1,2,4]triazolo[[1,5-a]pyridin-6-yl)cyclopentyl)acrylamide (0.53
g, 1.4 mmol) was separated by chiral preparative HPLC (Table 2,
Method 13) to give
N-((1S,3R)-3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c]][1,4]oxazin-2-yl)am-
ino)-[1,2,4]triazolo[1,5-a]pyridin-6-yl)cyclopentyl)acrylamide
(0.036 g, 7%, OR=negative). BTK enzyme IC.sub.50=A
[0495] The remaining mixture was repurified by chiral preparative
HPLC (Table 2, Method 14) to give
N-((1R,3S)-3-(8-((6,7-dihydro-4H-pyrazolo[5,1-c][1,4]oxazin-2-yl)amino)-[-
1,2,4]triazolo[1,5-a]pyridin-6-yl)cyclopentyl)acrylamide (0.033 g,
17%, OR=positive): LC/MS (Table 1, Method m) R.sub.t=1.42 min; MS
m/z 394 (M+H).sup.+. BTK enzyme IC.sub.50=A
Example #9:
N-((1S,3R)-3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclopentyl)acrylamide and
N-((1R,3S)-3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclopentyl)acrylamide
##STR00202##
[0496] Step A:
6-chloro-N-(5-morpholinopyridin-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-ami-
ne
##STR00203##
[0498] A reaction vial was charged with
8-bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine (0.500 g, 2.15
mmol, Example #1, Step A), 5-morpholinopyridin-2-amine (0.385 g,
2.15 mmol, ArkPharm), 1,4-dioxane (10 mL), Cs.sub.2CO.sub.3 (1.40
g, 4.30 mmol), Xantphos (0.0622 g, 0.108 mmol) and
Pd.sub.2(dha).sub.3 (0.098 g, 0.11 mmol), The reaction vial was
flushed with nitrogen, capped, stirred and heated to about
120.degree. C. overnight. The reaction was cooled to rt and then
diluted with DCM (100 mL) and water (80 mL). The organic layer was
separated, washed with water (80 mL), brine (100 mL), and dried
over Na.sub.2SO.sub.4. After concentrating the extract to dryness,
the product was purified via silica chromatography eluting with a
gradient of 20-50% EtOAc in petroleum ether to afford
6-chloro-N-(5-morpholinopyridin-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-ami-
ne (0.50 g, 70.3%): LC/MS (Table 1, Method r) R.sub.t=1.75 min; MS
m/z 331 (M+H).sup.+.
Step B: 4-((tert-butoxycarbonyl)amino)cyclopent-1-en-1-yl
trifluoromethanesulfonate
##STR00204##
[0500] LDA (37.6 mL, 75 mmol, 2 M) was added to a solution of
tert-butyl (3-oxocyclopentyl)carbamate (6 g, 30.1 mmol, ArkPharm)
in THF (2 mL) at about -78.degree. C. After about 20 min,
1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide
(11.83 g, 33.1 mmol) in THF (50 mL) was added and stirring was
continued for about a further 10 min before the cooling bath was
removed and the mixture allowed to reach rt. After about 2.5 h the
mixture was diluted with Et.sub.2O and washed sequentially with 1 N
aqueous sodium hydroxide and 1 N aqueous hydrochloric acid. The
solution was dried over MgSO.sub.4, concentrated under reduced
pressure and purified by flash chromatography to give the
intermediate 4-((tert-butoxycarbonyl)amino)cyclopent-1-en-1-yl
trifluoromethanesulfonate (1.4 g, 14%) and carried forward
immediately. A round bottom flask was charged with
4-((tert-butoxycarbonyl)amino)cyclopent-1-en-1-yl
trifluoromethanesulfonate (0.6 g, 1.811 mmol), DPPF (0.050 g, 0.091
mmol), and PdCl.sub.2(dppf)-CH.sub.2Cl.sub.2Adduct (0.074 g, 0.091
mmol) in 1,4-dioxane (10 mL) to give a brown suspension. Potassium
acetate (0.53 g, 5.4 mmol) and bis(pinacolato)diboron (0.460 g,
1.81 mmol) were added. The resulting mixture was heated at about
100.degree. C. overnight. The desired product was separated by
column to give tert-butyl
(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopent-3-en-1-yl)carba-
mate (0.36 g, 64%): LC/MS (Table 1, Method s) R.sub.t=1.85 min; MS
m/z 310 (M+H).sup.+.
Step C: tert-butyl
(3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-y-
l)cyclopent-3-en-1-yl)carbamate
##STR00205##
[0502] A mixture of
6-chloro-N-(5-morpholinopyridin-2-yl)-[1,2,4]triazolo[1,5-a]pyridin-8-ami-
ne (0.30 g, 0.91 mmol), tert-butyl
(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopent-3-en-1-yl)carba-
mate (0.393 g, 1.27 mmol), PdCl.sub.2(dppf) (0.066 g, 0.091 mmol)
and K.sub.2CO.sub.3 (0.91 mL, 2.72 mmol) in 1,4-dioxane (4 mL) was
heated to about 130.degree. C. for about 3 h in a 10 mL microwave
reaction vial. The mixture was cooled to rt and DCM (150 mL) was
added to the solution. The organic layer was washed with saturated
NaCl (3.times.50 mL). The organic layer was dried over
Na.sub.2SO.sub.4, filtered and concentrated. The crude product was
purified by Prep-TLC to give tert-butyl
(3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-y-
l)cyclopent-3-en-1-yl)carbamate (0.35 g, 66.3%) as a pale brown
solid: LC/MS (Table 1, Method m) R.sub.t=1.77 min; MS m/z 478
(M+H).sup.+.
Step D: tert-butyl
(3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-y-
l)cyclopentyl)carbamate
##STR00206##
[0504] A round bottom flask was charged with a solution of
tert-butyl
(3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-y-
l)cyclopent-3-en-1-yl)carbamate (0.350 g, 0.733 mmol) in MeOH, (100
mL) followed by the addition of 10% Pd/C (0.050 g, 0.47 mmol). The
suspension was stirred under an atmosphere of hydrogen at rt for
about 1 day. The suspension was filtered, and the filtrate was
concentrated under reduced pressure to give tert-butyl
(3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-y-
l)cyclopentyl)carbamate (0.310 g, 88%): LC/MS (Table 1, Method m)
R.sub.t=1.76 min; MS m/z 480 (M+H).sup.+.
Step E:
6-(3-aminocyclopentyl)-N-(5-morpholinopyridin-2-yl)-[1,2,4]triazol-
o[1,5-a]pyridin-8-amine hydrochloride
##STR00207##
[0506] A solution of 4 M HCl (7 mL, 28.0 mmol) in 1,4-dioxane was
added dropwise to a solution of tert-butyl
(3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-y-
l)cyclopentyl)carbamate (0.310 g, 0.646 mmol) in THF (14 mL). The
mixture was stirred at rt for about 3 h. The solvent was removed
under reduced pressure to give
6-(3-aminocyclopentyl)-N-(5-morpholinopyridin-2-yl)-[1,2,4]triazolo[1,5-a-
]pyridin-8-amine hydrochloride (0.22 g, 81%): LC/MS (Table 1,
Method n) R.sub.t=1.38 min; MS m/z 380 (M+H).sup.+.
Step F:
N-(3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-I]py-
ridin-6-yl)cyclopentyl)acrylamide
##STR00208##
[0508] TEA (0.073 mL, 0.53 mmol) was added dropwise into a
suspension of
6-(3-aminocyclopentyl)-N-(5-morpholinopyridin-2-yl)-[1,2,4]triazolo[1,5-a-
]pyridin-8-amine hydrochloride (0.219 g, 0.527 mmol) in DCM (8 mL)
at about 0.degree. C. The solution was stirred for about 10 min.
Then a solution of acryloyl chloride (0.062 g, 0.68 mmol) in DCM (1
mL) was added dropwise. The mixture was stirred for about 20 min
and the solvents were removed under reduced pressure. The crude
product was purified by prep-HPLC to get
N-(3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyridin-6-
-yl)cyclopentyl)acrylamide (0.17 g, 74%): LC/MS (Table 1, Method n)
R.sub.t=1.56 min; MS m/z 434 (M+H).sup.+.
Step G:
N-((1S,3R)-3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[-
1,5-a]pyridin-6-yl)cyclopentyl)acrylamide and
N-((1R,3S)-3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclopentyl)acrylamide
##STR00209##
[0510]
N-(3-(8-((5-Morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]pyr-
idin-6-yl)cyclopentyl)acrylamide (0.160 g, 0.369 mmol) was purified
by chiral preparative HPLC (Table 2, Method 15) to give
N-((1S,3R)-3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclopentyl)acrylamide (0.027 g, 17%, OR=negative). BTK
enzyme IC.sub.50=B
[0511] The remaining mixture (0.035 g, 0.081 mmol) was repurified
by chiral preparative HPLC (Table 2, Method 15) to give
N-((1R,3S)-3-(8-((5-morpholinopyridin-2-yl)amino)-[1,2,4]triazolo[1,5-a]p-
yridin-6-yl)cyclopentyl)acylamide (0.024 g, 69%, OR=positive):
LC/MS (Table 1, Method m) R.sub.t=1.49 min; MS m/z 434 (M+H).sup.+.
BTK enzyme IC.sub.50=A
Example #10:
(1R,3R)-3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyra-
zin-6-yl)cyclohexanol
##STR00210##
[0512] Step A:
6-bromo-N-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amin-
e
##STR00211##
[0514] A reaction vial was charged with
6,8-dibromo-[1,2,4]triazolo[1,5-a]pyrazine (10.38 g, 37.3 mmol,
ArkPharm),1-methyl-1H-pyrazol-4-amine (3.989 g, 41.1 mmol,
Astatech), DMF (100 mL), and. N-ethyl-N-isopropylpropan-2-anine
(12.92 nil, 74.7 mmol). The reaction vial was flushed with
nitrogen, and heated to about 100.degree. C. for about 90 minutes.
The reaction was cooled to rt and then added dropwise to stirring
water (200 mL) via an addition funnel. The resulting suspension was
filtered, washed with THF, then 1:1 EtOAc/Heptanes to afford
6-bromo-N-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amin-
e (9.8 g, 87%): LC/MS (Table 1, Method b) R.sub.t=1.66 min; MS m/z
295 (M+H).sup.+.
Step B:
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyraz-
in-6-yl)cyclohex-2-enone
##STR00212##
[0516] A mixture of
6-bromo-N-(1-methyl-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amin-
e (1 g, 3.40 mmol), tert-butyl
3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-2-enone
(0.755 g, 3.40 mmol), PdCl.sub.2(dppf) (0.239 g, 0.340 mmol) and
cesium carbonate (3.32 g, 10.20 mmol) was dissolved in 1,4-dioxane
(12 mL) and water (4 mL) and was heated to about 90.degree. C. for
about 16 h. The mixture was cooled to rt and water (5 mL) was added
to the solution. The suspension was filtered and washed with water
(10 mL), heptanes (9 mL), and ether (6 mL). The remaining solid was
purified via silicagel chromatography eluting with 0-10% MeOH/DCM
to afford
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)cyclohex-2-enone (0.9 g, 2.91 mmol, 86% yield) as a pale yellow
solid: LC/MS (Table 1, Method h) R.sub.t=1.47 min; MS m/z 310
(M+H).sup.+.
Step C:
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyraz-
in-6-yl)cyclohexanone
##STR00213##
[0518] A round bottom flask was charged with a solution of
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)cyclohex-2-enone (0.89 g, 2.88 mmol) in MeOH, (2 mL) followed by
the addition of 10% Pd/C (0.612 g, 0.575 mmol). The suspension was
stirred under an atmosphere of hydrogen at rt for about 18 h. The
suspension was filtered, and the filtrate was purified using
silicagel chromatography eluting with 0-8% MeOH/DCM to give
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)cyclohexanone (0.32 g, 1.02 mmol, 35% yield): LC/MS (Table 1,
Method h) R.sub.t=1.54 min; MS m/z 312 (M+H).sup.+.
Step D:
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyraz-
in-6-yl)cyclohexanone
##STR00214##
[0520] A round bottom flask was charged with a solution of
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)cyclohexanone (0.32 g, 1.028 mmol)) in THF (10.28 mL) and was
cooled to about -78.degree. C. L-Selectride (2.056 ml, 2.056 mmol)
was added dropwise to the reaction mixture and stirred at about
-78.degree. C. for 1 h then allowed to warm to rt over 4 h. The
reaction was quenched with the addition of aq. saturated ammonium
chloride (5 mL), then extracted with DCM (3.times.5 mL). The
combined organic layers were concentrated under reduced pressure.
The crude material was purified via silicagel chromatography
eluting with 0-10% MeOH/DCM to give
3-(8-((1-methyl-H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl)-
cyclohexanol (0.27 g, 84% yield) as an off-white solid: LC/MS
(Table 1, Method h) R.sub.t=1.45 min; MS m/z 314 (M+H).sup.+.
3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyrazin-6-yl-
)cyclohexanol (0.260 g, 0.369 mmol) was purified by chiral
preparative HPLC (Table 2, Method 16) to give
(1R,3R)-3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyra-
zin-6-yl)cyclohexanol (0.08 g, 24%, OR=negative). CSF-1R Enzyme
IC.sub.50=A.
Example #11:
(1R,3R)-3-(8-((1-methyl-1H-pyrazol-4-yl)amino)-[1,2,4]triazolo[1,5-a]pyra-
zin-6-yl)cyclohexanol
##STR00215##
[0521] Step A: 5-(cyclohex-1-en-1-yl)pyrazin-2-amine
##STR00216##
[0523] A round bottom flask was charged with 5-bromopyrazin-2-amine
(13.1 g, 75 mmol, ArkPharm),
2-(cyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
(15.7 g, 75 mmol, CombiBlocks) and potassium phosphate tribasic (32
g, 151 mmol) in a mixture of 1,4-dioxane (229 mL) and water (22.86
mL). The reaction mixture was degassed with nitrogen for about 10
min. while warming to about 50.degree. C. Pd(PPh.sub.3).sub.4 (2.62
g, 2.26 mmol) was added and the reaction mixture was heated to
about 80.degree. C. for about 5 h. Additional
Pd(PPh.sub.3).sub.4(0.7 mg) was added to the reaction mixture and
stirred at about 80.degree. C. overnight. The reaction was cooled
to ambient temperature and partitioned between EtOAc and brine
(2.times.50 mL). The combined organic portion was dried over
anhydrous MgSO.sub.4, and filtered through a plug of silicagel. The
solvent was concentrated in vacuo. The residue was taken up in
about 100 mL of EtOAc and 50 mL of Heptanes. The solid that
precipitated was collected to give
5-(cyclohex-1-en-1-yl)pyrazin-2-amine (8.6 g, 96% yield): LC/MS
(Table 1, Method h) R.sub.t=1.78 min; MS m/z 176 (M+H).sup.+.
Step B: 5-cyclohexylprazin-2-amine
##STR00217##
[0525] A round bottom flask was charged with 10% Pd/C carbon (3 g,
39.9 mmol) and wet EtOAc (5 mL). A solution of
5-(cyclohex-1-en-1-yl)pyrazin-2-amine (7 g, 39.9 mmol) in EtOH (256
mL) and acetic acid (10.24 mL) was added to the flask affixed with
a hydrogen balloon. The reaction was purged with hydrogen and
stirred for about 4 h at rt. The reaction mixture was filtered
through a pad of Celite.RTM. and the solvents were removed under
reduced pressure. The resulting solid was partitioned between EtOAc
and saturated aq. NaHCO.sub.3. The combined organic portion was
dried over anhydrous MgSO.sub.4, filtered, and concentrated in
vacuo. The residue was purified via silicagel chromatography
eluting with 0-80% EtOAc/Heptanes to give
5-cyclohexylpyrazin-2-amine (1.4 g, 19% yield): LC/MS (Table 1,
Method h) R.sub.t=1.78 min; MS m/z 178 (M+H).sup.+.
Step C: 3-bromo-5-cyclohexylpyrazin-2-amine
##STR00218##
[0527] In a round bottom flask, N-bromosuccinimide (1.6 g, 9.08
mmol) was added portionwise to a solution of
5-cyclohexylpyrazin-2-amine (1.4 g, 7.90 mmol) in DMF (15 mL). The
reaction stirred for about 2 h at rt. The reaction was quenched
with the addition of ice-cold water (30 mL). The reaction was
partitioned between EtOAc and saturated aq. NaHCO.sub.3, dried over
anhydrous MgSO4, filtered through a pad of silicagel, and
concentrated under reduced pressure to give
3-bromo-5-cyclohexylpyrazin-2-amine (1.42 g, 49% yield): LC/MS
(Table 1, Method h) R.sub.t=2.35 min; MS m/z 256, 258
(M+H).sup.+.
Step D:
(E)-N'-(3-bromo-5-cyclohexylpyrazin-2-yl)-N-hydroxyformimidamide
##STR00219##
[0529] In a round bottom flask, 3-bromo-5-cyclohexylpyrazin-2-amine
(1.2 g, 4.6 mmol) was dissolved in N,N-dimtheylformamide dimethyl
acetal (1.9 mL, 13.82 mmol). The mixture was heated to about
100.degree. C. for about 1 h. The solvent was concentrated in vacuo
to afford a crude residue. The crude,
(E)-N'-(3-bromo-5-cyclohexylpyrazin-2-yl)-N,N-dimethylformimidamid-
e (1.4 g, 4.61 mmol) was dissolved in MeOH (12 mL) and treated with
hydroxylamine hydrochloride (0.45 g, 6.45 mmol). The reaction
stirred for about 4 h at rt. The solvent was concentrated under
reduced pressure. 100 mL of water was added to the remaining
residue and the pH was adjusted to 9 with the addition of 1M aq.
NaOH. The solid that formed was filtered and collected to afford
(E)-N'-(3-bromo-5-cyclohexylpyrazin-2-yl)-N-hydroxyformimidamide
(1.2 g, 78% yield): LC/MS (Table 1, Method h) R.sub.t=2.42 min; MS
m/z=299, 301 (M+H).sup.+.
Step E: 8-bromo-6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazine
##STR00220##
[0531] To a solution of
(E)-N'-(3-bromo-5-cyclohexylpyrazin-2-yl)-N-hydroxyformimidamide
(1.2 g, 4.03 mmol) in MeCN (20 mL) was added trifluoroacetic
anhydride (0.85 mL, 6.05 mmol) and the mixture was stirred at rt
for about 3 h. The reaction mixture was partitioned between 1M NaOH
and EtOAc. The combined organic portion was dried over anhydrous
Na.sub.2SO.sub.4, filtered, and concentrated under reduced
pressure. The crude material was purified via silicagel
chromatography eluting with 0-100% EtOAc/Heptanes to afford
8-bromo-6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazine (0.75 g, 61%
yield): LC/MS (Table 1, Method h) R.sub.t=2.31 min; MS m/z=281, 283
(M+H).sup.+.
Step F:
1-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-p-
yrazol-1-yl)-2-methylpropan-2-ol
##STR00221##
[0533] A flask was charged with
8-bromo-6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazine (0.05 g, 0.178
mmol), 1-(4-amino-1H-pyrazol-1-yl)-2-methylpropan-2-ol (33.1 mg,
0.213 mmol, Preparation #3) and N-ethyl-N-isopropylpropan-2-amine
(0.046 g, 0.356 mmol) in DMF (2 mL). The reaction was stirred at
100.degree. C. for 12 hrs. The reaction was cooled to ambient
temperature and purified via prep-HPLC (Table 1, Method u) to give
1-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)-2-methylpropan-2-ol (0.03 g, 48% yield) as white solid.:
LC/MS (Table 1, Method v) R.sub.t=3.12 min; MS m/z=356 (M+H).sup.+.
CSF-1R Enzyme IC.sub.50=A
Example #12:
N-(6-cyclopentyl-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyraz-
olo [5,1-c][1,4]oxazin-2-amine
##STR00222##
[0534] Step A:
1-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)-2-methylpropan-2-ol
##STR00223##
[0536] To a solution of
N-(6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyrazolo[5-
,1-c][1,4]oxazin-2-amine (0.07 g, 0.241 mmol, Example 4, Step A) in
1,4-dioxane (2.4 mL) was added a 0.5 M THF solution of cyclopentyl
zinc bromide (2.9 mL, 1.45 mmol, Alfa Aesar) dropwise via syringe.
The reaction stirred under nitrogen for about 5 min. before the
addition of Pd(dppf)Cl.sub.2 (0.019 g, 0.024 mmol). The reaction
was heated to 85.degree. C. for 4 h. The reaction was cooled to
ambient temperature and was partitioned between saturated aq.
NaHCO.sub.3 and EtOAc (2.times.20 mL). The combined organic portion
was dried over anhydrous MgSO.sub.4, filtered and concentrated in
vacuo. The crude material was purified via silicagel
chromatography, eluting with 0-100% EtOAc/Heptanes to give
N-(6-cyclopentyl-[1,2,4]triazolo[1,5-a]pyridin-8-yl)-6,7-dihydro-4H-pyraz-
olo[5,1-c][1,4]oxazin-2-amine (0.036 g, 41% yield). LC/MS (Table 1,
Method h) R.sub.t=2.88 min; MS m/z=325(M+H).sup.+. CSF-1R Enzyme
IC.sub.50=A
Example #13:
1-(6-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyridin-8-yl)amino)pyridin-3-yl-
)piperidin-4-ol
##STR00224##
[0537] Step A: 1-(6-nitropyridin-3-yl)piperidin-4-ol
##STR00225##
[0539] A round bottom flask was charged with
5-bromo-2-nitropyridine (4.0 g, 19.7 mmol), 4-hydroxypiperidine
(2.4 g, 23.6 mmol), and potassium carbonate (5.5 g, 39.4 mmol) in
DMSO (5 mL). The reaction stirred at rt for about 20 h. The solid
that formed was filtered off, and the remaining filtrate was
concentrated under reduced pressure. The remaining residue was
triturated with DCM to afford 1-(6-nitropyridin-3-yl)piperidin-4-ol
(1.89 g, 43% yield). LC/MS (Table 1, Method h) R.sub.t=1.24 min; MS
m/z=224(M+H).sup.+.
Step B: 1-(6-aminopyridin-3-yl)piperidin-4-ol
##STR00226##
[0541] A stainless steel hydrogenation vessel was charged with
1-(6-nitropyridin-3-yl)piperidin-4-ol (1.89 g, 8.48 mmol) and 10%
palladium on carbon (0.541 g, 0.509 mmol) in MeOH (200 mL). The
mixture was shaken in a Parr hydrogenator pressurized with hydrogen
(about 30 psi) at ambient temperature. After about 1 h, the
reaction mixture was filtered through a pad of Celite.RTM., washing
with excess MeOH. The solvent was removed in vacuo to afford
1-(6-aminopyridin-3-yl)piperidin-4-ol (1.55 g, 95% yield). .sup.1H
NMR (400 MHz, DMSO-d6) .delta. 7.59 (dd, J=3.0, 0.7 Hz, 1H), 7.14
(dd, J=8.8, 3.0 Hz, 1H), 6.37 (dd, J=8.9, 0.7 Hz, 1H), 5.33 (bs,
2H), 4.62 (d, J=3.9 Hz, 1H), 3.54 (tq, J=8.3, 3.9 Hz, 1H),
3.24-3.13 (m, 2H), 2.62 (ddd, J=12.5, 10.0, 2.9 Hz, 2H), 1.85-1.74
(m, 2H), 1.54-1.42 (m, 2H).
Step C:
1-(6-((6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)amino)pyridin-3-
-yl)piperidin-4-ol
##STR00227##
[0543] A round bottom flask was charged with
8-bromo-6-chloro-[1,2,4]triazolo[1,5-a]pyridine (1.1 g, 4.70 mmol,
Example 1, Step A), 1-(6-aminopyridin-3-yl)piperidin-4-ol (1.0 g,
5.17 mmol), cesium carbonate (3.1 g, 9.41 mmol), and Xantphos (0.16
g, 0.282 mmol, Strem) in 1,4-dioxane (50 mL). The reaction mixture
was sparged with nitrogen for 15 minutes, before the addition of
Pd(OAc).sub.2 (0.03 g, 0.14 mmol). The reaction was heated to about
80.degree. C. for about 16 h. The reaction was cooled to ambient
temperature and the solvent was removed in vacuo. The crude
material was purified via silicagel chromatography eluting with
10-100% EtOAc/MeOH to afford
1-(6-((6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)amino)pyridin-3-yl)pip-
eridin-4-ol (1.1 g, 64% yield). LC/MS (Table 1, Method h)
R.sub.t=1.75 min; MS m/z=345 (M+H).sup.+.
Step D:
1-(6-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyridin-8-yl)amino)pyrid-
in-3-yl)piperidin-4-ol
##STR00228##
[0545] A reaction vial was charged with
1-(6-((6-chloro-[1,2,4]triazolo[1,5-a]pyridin-8-yl)amino)pyridin-3-yl)pip-
eridin-4-ol (0.25 g, 0.73 mmol) in 1,4-dioxane (7.2 mL). The vial
was sparged with nitrogen before the addition of 0.5 M solution of
cyclohexyl zinc bromide in THF (11.6 mL, 5.80 mmol) and
Pd(dppf)Cl.sub.2 (0.05 g, 0.07 mmol). The reaction was heated to
about 85.degree. C. for about 1 h. The reaction was cooled to
ambient temperature and partitioned between EtOAc and water. The
combined organic portion was washed with 1N aq. NaOH, dried over
MgSO.sub.4, filtered, and concentrated under reduced pressure. The
crude material was purified via Preparative HPLC (Table 1, Method
y) to give
1-(6-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyridin-8-yl)amino)pyridin-3-yl-
)piperidin-4-ol (0.01 g, 11% yield). LC/MS (Table 1, Method h)
R.sub.t=2.21 min; MS m/z=393(M+H).sup.+. CSF-1R Enzyme
IC.sub.50=A
Example #14:
6-cyclohexyl-N-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazo-
lo[1,5-a]pyrazin-8-amine
##STR00229##
[0546] Step A:
6-cyclohexyl-N-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazo-
lo[1,5-a]pyrazin-8-amine
##STR00230##
[0548] A round bottom flask was charged with
6-cyclohexyl-N-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a-
]pyrazin-8-amine, hydrochloric acid (0.40 g, 0.99 mmol, Table
D1.1), paraformaldehyde (0.06 g, 1.98 mmol), acetic acid (0.17 mL,
2.98 mmol), and sodium triacetoxyhydroborate (0.21 g, 0.99 mmol) in
MeOH (9.93 mL). The reaction was heated to about 50.degree. C. for
about 18 h. The reaction was cooled to ambient temperature and the
solvent was removed in vacuo. The residue was partitioned between
saturated aq. NaHCO.sub.3 (20 mL) and DCM (2.times.20 mL). The
combined organic portion was dried over MgSO4, filtered, and
concentrated under reduced pressure. The crude material was
purified via preparative HPLC (Table 2, Method 20) to give
6-cyclohexyl-N-(1-(1-methylpiperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazo-
lo[1,5-a]pyrazin-8-amine (0.032 g, 9% yield). LC/MS (Table 1,
Method h) R.sub.t=1.65 min; MS m/z=381(M+H).sup.+. CSF-1R Enzyme
IC.sub.50=A
Example #15:
3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)propan-1-ol
##STR00231##
[0549] Step A:
6-bromo-N-(1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amine
##STR00232##
[0551] To a solution of 6,8-dibromo-[1,2,4]triazolo[1,5-a]pyrazine
(0.989 g, 3.56 mmol, ArkPharm) in DMF (18 mL) was added DIEA (1.9
ml, 10.6 mmol) and 1H-pyrazol-4-amine (0.44 g, 5.34 mmol, Combi
Blocks). The reaction was heated to about 95.degree. C. for about 3
h. The reaction was cooled to ambient temperature and the solvent
was removed in vacuo. The remaining residue was suspended in
H.sub.2O (20 mL) and stirred overnight at rt. The resulting solid
was filtered, resuspended in EtOAc (20 mL), and filtered to afford
6-bromo-N-(1H-pyrazol-4-yl)-1,2,4/triazolo[1,5-a]pyrazin-8-amine
(0.9 g, 90% yield) as a grey solid. LC/MS (Table 1, Method h)
R.sub.t=1.52 min; MS m/z=280 (M+H).sup.+.
Step B: tert-butyl
4-((6-bromo-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazole-1-carbo-
xylate
##STR00233##
[0553] To a solution of
6-bromo-N-(1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amine
(0.79 g, 2.8 mmol), N,N-dimethylpyridin-4-amine (0.03 g, 0.28
mmol), and TEA (0.59 mL, 4.26 mmol) in DCM (19 mL) was added
Boc.sub.2O (0.62 g, 2.84 mmol). The mixture was stirred at rt for
about 4 h. The solvent was removed in vacuo. The remaining residue
was purified via silicagel chromatography eluting with MeOH/DCM
(0-3%) to afford tert-butyl
4-((6-bromo-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazole-1-carbo-
xylate (0.72 g, 66% yield) as a white solid. LC/MS (Table 1, Method
h) R.sub.t=2.23 min; MS m/z=378 (M-H).sup.-.
Step C:
6-cyclohexyl-N-(1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-a-
mine
##STR00234##
[0555] A reaction vial was charged with tert-butyl
4-((6-bromo-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazole-1-carbo-
xylate (0.72 g, 1.88 mmol) in 1,4-dioxane (7.5 mL). The vial was
sparged with nitrogen for about 5 min. before the addition of 0.5 M
solution of cyclohexyl zinc bromide in THF (22 mL, 11.33 mmol), and
Pd(dppf)Cl.sub.2 (0.14 g, 0.19 mmol). The reaction was heated to
about 75.degree. C. for about 30 minutes. The reaction was cooled
to ambient temperature and was partitioned between DCM (3.times.40
mL) and saturated aq. NaHCO.sub.3 (40 mL). The combined organic
portion was dried over MgSO.sub.4, filtered, and concentrated under
reduced pressure. The crude material was purified via silicagel
chromatography eluting with MeOH/DCM (0-10%) to give
6-cyclohexyl-N-(1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amine
(0.41 g, 77% yield). LC/MS (Table 1, Method h) R.sub.t=2.01 min; MS
m/z=284 (M+H).sup.+.
Step D:
3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-p-
yrazol-1-yl)propan-1-ol
##STR00235##
[0557] A reaction vial was charged with
6-cyclohexyl-N-(1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amine
(0.12 g, 0.43 mmol), K.sub.2CO.sub.3 (0.08 g, 0.64 mmol), and
3-iodopropan-1-ol (1 mL, 0.86 mmol) in DMF (4.5 mL). The reaction
was heated to about 90.degree. C. for about 16 h. An additional 2
equivalents of 3-iodopropan-1-ol (2 mL, 0.86 mmol) was then added
to the reaction and continued to stir at about 90.degree. C. for
about 4 h. The reaction was cooled to ambient temperature and was
purified via preparative HPLC (Table 1, Method aa) to give
3-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)propan-1-ol (0.04 g, 25% yield) as an off-white solid. LC/MS
(Table 1, Method h) R.sub.t=1.97 min; MS m/z=342 (M+H).sup.+.
CSF-1R Enzyme IC.sub.50=A
Example #16:
cis-4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyra-
zol-1-yl)cyclohexanecarboxylic acid
##STR00236##
[0559] To a solution of cis-ethyl
4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)cyclohexanecarboxylate (0.05 g, 0.12 mmol, Table A.1.8) in
MeOH (0.6 mL) was added 1N aq. NaOH (0.25 mL, 0.25 mmol). The
reaction as stirred at rt for about 16 h. The solvent was
concentrated under reduced pressure, and the remaining residue was
purified via preparative HPLC (Table 1, Method ab) to afford
cis-4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyra-
zol-1-yl)cyclohexanecarboxylic acid (0.018 g, 35% yield) as an
off-white solid. LC/MS (Table 1, Method h) R.sub.t=2.27 min; MS
m/z=410 (M+H).sup.+. CSF-1R Enzyme IC.sub.50=A
Example #17:
6-cyclohexyl-N-(1-((2R,4s,6S)-2,6-dimethylpiperidin-4-yl)-1H-pyrazol-4-yl-
)-[1,2,4]triazolo[1,5-a]pyrazin-8-amine
##STR00237##
[0560] Step A: tert-butyl
4-(4-((6-bromo-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)-
-2,6-dimethylpiperidine-1-carboxylate
##STR00238##
[0562] To a solution of 6,8-dibromo-[1,2,4]triazolo[1,5-a]pyrazine
(1.54 g, 5.54 mmol, ArkPharm) in DMF (20 mL) was added tert-butyl
4-(4-amino-1H-pyrazol-1-yl)-2,6-dimethylpiperidine-1-carboxylate
(1.79 g, 6.10 mmol, Preparation #10), and DIEA (1.16 ml, 6.65
mmol). The reaction mixture was heated to about 100.degree. C. for
about 14 h. The reaction cooled to ambient temperature and was
partitioned between water (40 mL) and EtOAc (3.times.40 mL). The
combined organic portion was dried over Na.sub.2SO.sub.4, filtered,
and concentrated under reduced pressure. The residue was purified
via silicgel chromatography eluting with EtOAc/Petroleum ether
(0-10%) to afford tert-butyl
4-(4-((6-bromo-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)-
-2,6-dimethylpiperidine-1-carboxylate (2.5 g, 92% yield) as a
yellow solid. LC/MS (Table 1, Method w) R.sub.t=1.50 min; MS
m/z=492 (M+H).sup.+.
Step B: (2R,4s,6S)-tert-butyl
4-(4-((6-(cyclohex-1-en-1-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1-
H-pyrazol-1-yl)-2,6-dimethylpiperidine-1-carboxylate
##STR00239##
[0564] A round bottom flask was charged with tert-butyl
4-(4-((6-bromo-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol-1-yl)-
-2,6-dimethylpiperidine-1-carboxylate (2.4 g, 4.88 mmol),
2-(cyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
(1.220 g, 5.86 mmol, ArkPharm), Na.sub.2CO.sub.3 (1.5 g, 14.6
mmol), and Pd(Ph.sub.3P).sub.4 (0.56 g, 0.488 mmol) in DMF (12 mL)
and Water (9 mL). The reaction was heated to about 80.degree. C.
for about 14 h. The reaction cooled to ambient temperature and was
partitioned between water (40 mL) and EtOAc (3.times.50 mL). The
combined organic portion was dried over Na.sub.2SO.sub.4, filtered,
and concentrated under reduced pressure. The residue was purified
via silicgel chromatography eluting with EtOAc/Petroleum ether
(0-10%) to afford racemic product. The racemate was subjected to
preparative chiral SFC (Table 2, Method 19) to give
(2R,4s,6S)-tert-butyl
4-(4-((6-(cyclohex-1-en-1-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1-
H-pyrazol-1-yl)-2,6-dimethylpiperidine-1-carboxylate (0.71 g, 29%
yield) as a white solid. LC/MS (Table 2, Method 19) R.sub.t=3.12
min; MS m/z=493 (M+H).sup.+.
Step C: (2R,4r,6S)-tert-butyl
4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)-2,6-dimethylpiperidine-1-carboxylate
##STR00240##
[0566] To a solution of (2R,6S)-tert-butyl
4-(4-((6-(cyclohex-1-en-1-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1-
H-pyrazol-1-yl)-2,6-dimethylpiperidine-1-carboxylate (200 mg, 0.41
mmol) in a mixture of MeOH (5 mL), THF (5 mL), and AcOH (0.25 mL)
was added 10% Pd/C (216 mg, 2.03 mmol). The reaction was stirred at
rt under an atmosphere of hydrogen for about 16 hrs. The reaction
mixture was filtered through a pad of Celite.RTM. and concentrated
under reduced pressure to afford (2R,6S)-tert-butyl
4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)-2,6-dimethylpiperidine-1-carboxylate (170 mg, 85% yield) as a
white solid.
[0567] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.61 (s, 1H), 8.25
(s, 1H), 8.13 (s, 1H), 7.93 (s, 1H), 7.77 (s, 1H), 4.77-4.72 (m,
1H), 4.61-4.56 (m, 2H), 2.67-2.61 (m, 1H), 2.17-2.04 (m, 8H),
1.91-1.95 (m, 4H), 1.51 (s, 9H), 1.47 (s, 3H), 1.44 (s, 3H),
1.37-1.29 (m, 2H).
Step D:
6-cyclohexyl-N-(1-((2R,4s,6S)-2,6-dimethylpiperidin-4-yl)-1H-pyraz-
ol-4-yl)-[1,2,4]triazolo[1,5-a]pyrazin-8-amine
##STR00241##
[0569] To a solution of (2R,4s,6S)-tert-butyl
4-(4-((6-cyclohexyl-[1,2,4]triazolo[1,5-a]pyrazin-8-yl)amino)-1H-pyrazol--
1-yl)-2,6-dimethylpiperidine-1-carboxylate (170 mg, 0.344 mmol) in
DCM (15 mL) was added TFA (5 mL, 64.9 mmol). The reaction mixture
was stirred at rt for about 2 h. The reaction was stirred at rt for
about 16 h. The solvent was concentrated under reduced pressure,
and the remaining residue was partitioned between DCM and saturated
aq. NaHCO.sub.3. The organic portion was dried over anhydrous
Na2SO4, filtered, and concentrated under reduced pressure. The
resulting solid was recrystallized from EtOAc (2 mL) to afford
6-cyclohexyl-N-(1-((2R,4s,
6S)-2,6-dimethylpiperidin-4-yl)-1H-pyrazol-4-yl)-[1,2,4]triazolo[1,5-a]py-
razin-8-amine (52 mg, 39% yield) as a white solid. LC/MS (Table 1,
Method w) R.sub.t=2.69 min; MS m/z=395 (M+H).sup.+. CSF-1R Enzyme
IC.sub.50=A
* * * * *