U.S. patent application number 15/437143 was filed with the patent office on 2017-06-08 for 2-aminopyrazine derivatives as csf-1 r kinase inhibitors.
The applicant listed for this patent is Chroma Therapeutics Ltd. Invention is credited to Stephen John Davies, David F. C. Moffat, Carl Leslie North, Stephane Pintat.
Application Number | 20170157118 15/437143 |
Document ID | / |
Family ID | 46704225 |
Filed Date | 2017-06-08 |
United States Patent
Application |
20170157118 |
Kind Code |
A1 |
Davies; Stephen John ; et
al. |
June 8, 2017 |
2-AMINOPYRAZINE DERIVATIVES AS CSF-1 R KINASE INHIBITORS
Abstract
The present invention provides a compound of formula (I) or a
pharmaceutically acceptable salt, N-oxide, hydrate or solvate
thereof: wherein: ring A, R.sup.1, R.sup.2, n, X, V, W, Z, ring B,
[Linker] and R areas defined herein. The compounds are useful as
inhibitors of CSF-1R kinase. The compounds can thus be used in
medicine. ##STR00001##
Inventors: |
Davies; Stephen John;
(Abingdon, GB) ; Pintat; Stephane; (Abingdon,
GB) ; North; Carl Leslie; (Abingdon, GB) ;
Moffat; David F. C.; (Abingdon, GB) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Chroma Therapeutics Ltd |
Abingdon |
|
GB |
|
|
Family ID: |
46704225 |
Appl. No.: |
15/437143 |
Filed: |
February 20, 2017 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
14411024 |
Dec 23, 2014 |
9604940 |
|
|
PCT/GB2013/051693 |
Jun 26, 2013 |
|
|
|
15437143 |
|
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 1/00 20180101; A61P
1/04 20180101; A61P 35/00 20180101; A61P 43/00 20180101; A61P 9/10
20180101; A61P 11/08 20180101; A61P 9/00 20180101; C07D 401/12
20130101; C07D 403/12 20130101; A61P 37/02 20180101; A61P 25/28
20180101; A61P 19/02 20180101; A61P 3/00 20180101; A61P 17/06
20180101; A61P 31/00 20180101; A61P 37/08 20180101; C07D 401/14
20130101; C07D 403/10 20130101; A61P 7/00 20180101; C07D 241/20
20130101; A61P 3/10 20180101; A61P 29/00 20180101; A61P 11/06
20180101; C07D 403/04 20130101; A61P 25/00 20180101; A61P 17/00
20180101 |
International
Class: |
A61K 31/497 20060101
A61K031/497 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 26, 2012 |
GB |
1211310.6 |
Claims
1. A compound which is an amino acid or amino acid ester of formula
(I) or a salt, N-oxide, hydrate or solvate thereof: ##STR00136##
wherein: ring A is a C.sub.6-12 aryl, 5- to 12-membered
heterocyclyl or C.sub.3-7 carbocyclyl ring; R.sup.1 and R.sup.2
independently represent hydrogen, halogen or an unsubstituted group
selected from C.sub.1-4 alkyl, C.sub.2-4 alkenyl, C.sub.1-4 alkoxy,
C.sub.2-4 alkenyloxy, C.sub.1-4 haloalkyl, C.sub.2-4 haloalkenyl,
C.sub.1-4 haloalkoxy, C.sub.2-4 haloalkenyloxy, hydroxyl,
--SR.sup.3, cyano, nitro, C.sub.1-4 hydroxyalkyl and
--NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4 are the same
or different and represent hydrogen or unsubstituted C.sub.1-2
alkyl; n is 0 or 1; X is NH or O; V and W independently represent
--N.dbd. or --C(Z).dbd.; each Z is the same or different and
represents hydrogen, fluoro, chloro, bromo, iodo, cyano,
unsubstituted C.sub.1-3 fluoroalkyl or unsubstituted
C.sub.1-3alkyl; ring B is a C.sub.6-12 aryl, 5- to 12-membered
heterocyclyl or C.sub.3-7 carbocyclyl ring; [Linker] represents a
group of formula --(CH.sub.2).sub.m--X.sup.1-L.sup.1-Y.sup.1--
wherein: m is 0, 1, 2 or 3; X.sup.1 represents a bond, --O--,
--S--, --NR.sup.7--, --C(.dbd.O)--, --C(.dbd.O)O--, --OC(.dbd.O)--,
--S(.dbd.O).sub.2--; --NR.sup.5C(.dbd.O)--, --C(.dbd.O)NR.sup.5--,
--NR.sup.5C(.dbd.O)NR.sup.6--, --NR.sup.5S(.dbd.O).sub.2--, or
--S(.dbd.O).sub.2NR.sup.5-- wherein R.sup.5 and R.sup.6 are
independently hydrogen or C.sub.1-6 alkyl and R.sup.7 represents
hydrogen, unsubstituted C.sub.1-4 alkyl or --C(.dbd.O)CH.sub.3;
L.sup.1 represents a divalent radical of formula
-(Alk.sup.1)-(Q).sub.y-(Het).sub.w-(Alk.sup.2).sub.z- or
-(Alk.sup.1).sub.x-(Het).sub.w-(Q).sub.y-(Alk.sup.2).sub.z- wherein
x, y, w and z are independently 0 or 1; Q represents a divalent
C.sub.6-12 aryl, 5- to 12-membered heterocyclyl or C.sub.3-7
carbocyclyl ring; Het represents --O--, --S-- or --NR.sup.7--
wherein R.sup.7 represents hydrogen, unsubstituted C.sub.1-4 alkyl
or --C(.dbd.O)CH.sub.3; Alk.sup.1 and Alk.sup.2 independently
represent divalent C.sub.3-7 cycloalkyl radicals, or straight or
branched, C.sub.1-6 alkylene, C.sub.2-6 alkenylene, or C.sub.2-6
alkynylene radicals; Y.sup.1 represents a bond, --O--, --S--,
--NR.sup.7--, --C(.dbd.O)--, --C(.dbd.O)O--, --OC(.dbd.O)--,
--S(.dbd.O).sub.2--, --NR.sup.5C(.dbd.O)--, --C(.dbd.O)NR.sup.5--,
--NR.sup.5C(.dbd.O)NR.sup.6--, --NR.sup.5S(.dbd.O).sub.2--, or
--S(.dbd.O).sub.2NR.sup.5--, wherein R.sup.5 and R.sup.6 are
independently hydrogen or C.sub.1-6 alkyl and R.sup.7 represents
hydrogen, unsubstituted C.sub.1-4 alkyl or --C(.dbd.O)CH.sub.3; R
represents a group of formula (X) or (Y): ##STR00137## in which:
R.sup.8 is a group --COOH or an ester group which is hydrolysable
by one or more intracellular carboxylesterase enzymes to a --COOH
group; R.sup.9 and R.sup.10 are the same or different and each
represents the .alpha.-substituent of a natural or non-natural
.alpha.-amino acid, or R.sup.9 and R.sup.10, taken together with
the carbon to which they are attached, form a 3- to 6-membered
saturated spiro cycloalkyl or heterocyclyl ring; R.sup.11
represents a hydrogen atom or a C.sub.1-4 alkyl group; ring D is a
5- to 7-membered saturated heterocyclyl group having at least one
nitrogen atom in the ring, wherein R.sup.8 and R.sup.11 are linked
to a ring carbon adjacent to a ring nitrogen, and wherein the bond
shown intersected by a wavy line may be from a nitrogen atom or a
carbon atom in ring D; wherein when R is (X), [Linker] is not
connected to (X) via an O, N or S atom; and wherein, unless
otherwise stated: any alkyl, alkenyl and alkynyl groups and
moieties in R.sup.5, R.sup.6, R.sup.8, R.sup.9, R.sup.10, R.sup.11,
Alk.sup.1 and Alk.sup.2 are the same or different and are each
unsubstituted or substituted with 1, 2 or 3 unsubstituted
substituents which are the same or different and are selected from
halogen atoms and C.sub.1-4 alkoxy, C.sub.2-4 alkenyloxy, C.sub.1-4
haloalkoxy, C.sub.2-4 haloalkenyloxy, hydroxyl, --SR.sup.3, cyano,
nitro and --NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4 are
the same or different and represent hydrogen or unsubstituted
C.sub.1-2 alkyl; any aryl, heterocyclyl, cycloalkyl and carbocyclyl
groups and moieties in rings A, B, Q, D, Alk.sup.1 and Alk.sup.2
and the ring formed by R.sup.9 and R.sup.10 are the same or
different and are each unsubstituted or substituted by 1, 2, 3 or 4
unsubstituted substituents selected from halogen atoms, and cyano,
nitro, C.sub.1-4 alkyl, C.sub.1-4 alkoxy, C.sub.2-4 alkenyl,
C.sub.2-4 alkenyloxy, C.sub.1-4 haloalkyl, C.sub.2-4 haloalkenyl,
C.sub.1-4 haloalkoxy, C.sub.2-4 haloalkenyloxy, hydroxyl, C.sub.1-4
hydroxyalkyl, --SR.sup.3 and --NR.sup.3R.sup.4 groups wherein each
R.sup.3 and R.sup.4 is the same or different and represents
hydrogen or unsubstituted C.sub.1-2 alkyl, or from substituents of
formula --COOR.sup.12, --COR.sup.12, --SO.sub.2R.sup.12,
--CONR.sup.12R.sup.13, --SO.sub.2NR.sup.12R.sup.13,
--OCONR.sup.12R.sup.13, --NR.sup.12COR.sup.13,
--NR.sup.12COOR.sup.13, --NR.sup.12SO.sub.2R.sup.13,
--NR.sup.12SO.sub.2OR.sup.13 or --NR.sup.12CONR.sup.12R.sup.13
wherein R.sup.12 and R.sup.13 are the same or different and
represent hydrogen or unsubstituted C.sub.1-4 alkyl, or R.sup.12
and R.sup.13 when attached to the same nitrogen atom form a
non-fused 5- or 6-membered heterocyclyl group, with the proviso
that the compound is not:
4-(4-{6-Amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-p-
yrazol-1-yl)-pyrrolidine-2-carboxylic acid methyl ester.
2. A compound as claimed in claim 1, wherein X is NH.
3. A compound as claimed in claim 1 or claim 2, wherein ring A and
ring B independently represent phenyl or a 5- to 6-membered
heterocyclyl group, said ring A or ring B group being unsubstituted
or substituted by 1, 2 or 3 substituents which are the same or
different and are selected from halogen atoms and unsubstituted
C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, C.sub.1-4 haloalkyl,
C.sub.1-4 haloalkoxy, C.sub.1-4 hydroxyalkyl, cyano, nitro,
--SR.sup.3 and --NR.sup.3R.sup.4 groups, wherein R.sup.3 and
R.sup.4 are the same or different and represent hydrogen or
unsubstituted C.sub.1-2 alkyl.
4. A compound as claimed in any one of claims 1 to 3, wherein ring
B is a 1,4-phenylene, 1,3-phenylene or pyridinyl group.
5. A compound as claimed in any one of claims 1 to 4, wherein
R.sup.1 and R.sup.2 independently represent hydrogen, halogen or
unsubstituted C.sub.1-4 alkyl.
6. A compound as claimed in any one of claims 1 to 5, wherein
R.sup.8 is a group --COOH or an ester group of formula
--(C.dbd.O)OR.sup.14 wherein R.sup.14 is
R.sup.15R.sup.16R.sup.17C-- wherein (i) R.sup.15 is hydrogen,
fluorine or optionally substituted
C.sub.1-3alkyl-(Z.sup.1).sub.a--[(C.sub.1-C.sub.3)alkyl].sub.b- or
C.sub.2-3alkenyl-(Z.sup.1).sub.a--[C.sub.1-3 alkyl].sub.b- wherein
a and b are independently 0 or 1 and Z.sup.1 is --O--, --S--, or
--NR.sup.18-- wherein R.sup.18 is hydrogen or C.sub.1-3 alkyl; and
R.sup.16 and R.sup.17 are independently hydrogen or C.sub.1-3
alkyl-; (ii) R.sup.15 is hydrogen or optionally substituted
R.sup.19R.sup.20N--C.sub.1-3alkyl--wherein R.sup.19 is hydrogen or
C.sub.1-3alkyl and R.sup.20 is hydrogen or C.sub.1-3alkyl; or
R.sup.19 and R.sup.20 together with the nitrogen to which they are
attached form an optionally substituted monocyclic heterocyclic
ring of 5- or 6-ring atoms or bicyclic heterocyclic ring system of
8 to 10 ring atoms, and R.sup.16 and R.sup.17 are independently
hydrogen or C.sub.1-3alkyl-; or (iii) R.sup.15 and R.sup.16 taken
together with the carbon to which they are attached form an
optionally substituted monocyclic carbocyclic ring of from 3 to 7
ring atoms or bicyclic carbocyclic ring system of 8 to 10 ring
atoms, and R.sup.17 is hydrogen; or (iv) R.sup.15 and R.sup.15
taken together with the carbon to which they are attached form an
optionally substituted monocyclic heterocyclic ring of from 3 to 7
ring atoms wherein at least one ring atom is selected from --O--,
--S--, or --NR.sup.18-- wherein R.sup.18 is hydrogen or
C.sub.1-3alkyl, or a bicyclic heterocyclic ring system of 8 to 10
ring atoms wherein at least one ring atom is selected from --O--,
--S--, or --NR.sup.18-- wherein R.sup.18 is hydrogen or
C.sub.1-3alkyl, and R.sup.17 is hydrogen.
7. A compound as claimed in claim 6, wherein either R.sup.15 is
hydrogen or
C.sub.1-3alkyl-(Z.sup.1).sub.a--[(C.sub.1-C.sub.3)alkyl].sub.b-
wherein a and b are independently 0 or 1 and Z.sup.1 is --O--,
--S--, or --NH-- and R.sup.16 and R.sup.17 are independently
hydrogen or C.sub.1-3 alkyl-; or R.sup.15 and R.sup.15 taken
together with the carbon to which they are attached form a 3- to
7-membered cycloalkyl ring and R.sup.17 is hydrogen.
8. A compound as claimed in claim 6, wherein either R.sup.14 is
cyclopentyl or t-butyl.
9. A compound as claimed in any one of the preceding claims,
wherein (i) R.sup.9 and R.sup.10 are side chains of natural amino
acids, (ii) one of R.sup.9 and R.sup.10 is hydrogen or
unsubstituted C.sub.1-4 alkyl and the other is an unsubstituted
C.sub.1-6 alkyl group or a C.sub.1-6 alkyl group substituted with a
C.sub.1-4 alkoxy group, or (iii) R.sup.9 and R.sup.10, taken
together with the carbon to which they are attached, form a
saturated spiro cyclobutyl ring.
10. A compound as claimed in any one of the preceding claims,
wherein R represents a group of formula (Y1): ##STR00138## wherein
ring D is a is a 5- to 7-membered saturated heterocyclyl group
having at least two nitrogen atoms in the ring, wherein R.sup.8 and
R.sup.11 are linked to a ring carbon adjacent to a ring nitrogen,
and wherein the bond shown intersected by a wavy line is from a
nitrogen atom in ring D.
11. A compound as claimed in any one of claims 1 to 9, wherein R
represents a group of formula (Y2): ##STR00139## wherein ring D is
a is a 5- to 7-membered saturated heterocyclyl group having at
least one nitrogen atom in the ring, wherein R.sup.8 and R.sup.11
are linked to a ring carbon adjacent to a ring nitrogen, and
wherein the bond shown intersected by a wavy line is from a carbon
atom in ring D.
12. A compound according to claim 11, wherein [Linker] represents
--(CH.sub.2).sub.m--X.sup.1-(Alk.sup.1).sub.x-Y, wherein m is 0, 1,
2 or 3, x is 0 or 1, Alk.sup.1 is an unsubstituted C.sub.1-3
alkylene group, and X.sup.1 and Y.sup.1 independently represent a
bond, --O--, --S--, --NR.sup.7--, --C(.dbd.O)-- or
--C(.dbd.O)NR.sup.5--, wherein R.sup.5 is hydrogen or C.sub.1-4
alkyl and R.sup.7 represents hydrogen, unsubstituted C.sub.1-4
alkyl or --C(.dbd.O)CH.sub.3.
13. A compound as claimed in any one of claims 1 to 3 or 5 to 12,
wherein: ring A represents phenyl or a 5- to 6-membered
heterocyclyl group, said ring A group being unsubstituted or
substituted by 1, 2 or 3 substituents which are the same or
different and are selected from halogen atoms and unsubstituted
C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, C.sub.1-4 haloalkyl,
C.sub.1-4 haloalkoxy, C.sub.1-4 hydroxyalkyl, cyano, nitro,
--SR.sup.3 and --NR.sup.3R.sup.4 groups, wherein R.sup.3 and
R.sup.4 are the same or different and represent hydrogen or
unsubstituted C.sub.1-2 alkyl; R.sup.1 and R.sup.2 each represent
hydrogen; n is 1; X is NH; V is --N.dbd.; W is --C(Z).dbd., wherein
Z represents hydrogen, fluoro, chloro, or unsubstituted C.sub.1-3
alkyl, preferably hydrogen; and ring B represents phenyl or 5- to
6-membered heterocyclyl group, said ring B group being
unsubstituted or substituted by 1, 2 or 3 substituents which are
the same or different and are selected from halogen atoms and
unsubstituted C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl,
C.sub.1-4 haloalkyl, C.sub.1-4 haloalkoxy, C.sub.1-4 hydroxyalkyl,
cyano, nitro, --SR.sup.3 and --NR.sup.3R.sup.4 groups, wherein
R.sup.3 and R.sup.4 are the same or different and represent
hydrogen or unsubstituted C.sub.1-2 alkyl.
14. A compound according to claim 13, wherein the group [Linker]-R
is one of (a) to (c): (a) R represents a group of formula (X);
R.sup.8 is a group --COOH or an ester group of formula
--(C.dbd.O)OR.sup.14 wherein R.sup.14 is
R.sup.15R.sup.16R.sup.17C-- wherein either R.sup.15 is hydrogen or
C.sub.1-3alkyl-(Z.sup.1).sub.a--[(C.sub.1-C.sub.3)alkyl].sub.b-
wherein a and b are independently 0 or 1 and Z.sup.1 is --O--,
--S--, or --NH-- and R.sup.16 and R.sup.17 are independently
hydrogen or C.sub.1-3 alkyl-; or R.sup.15 and R.sup.16 taken
together with the carbon to which they are attached form a 3- to
7-membered cycloalkyl ring and R.sup.17 is hydrogen; R.sup.9 and
R.sup.10 are side chains of natural amino acids or one of R.sup.9
and R.sup.10 is hydrogen or unsubstituted C.sub.1-4 alkyl and the
other is an unsubstituted C.sub.1-6 alkyl group; R.sup.11
represents a hydrogen atom or an unsubstituted C.sub.1-2 alkyl
group; [Linker] represents --(CH.sub.2).sub.m--X.sup.1-(Alk.sup.1)-
or --(CH.sub.2).sub.v--, wherein v is 1 or 2, m is 0, 1, 2 or 3;
Alk.sup.1 is an unsubstituted C.sub.1-3 alkylene group and X.sup.1
is a bond, --O--, --S--, --NR.sup.7--, --C(.dbd.O)-- or
--C(.dbd.O)NR.sup.5--, wherein R.sup.5 is hydrogen or C.sub.1-4
alkyl and R.sup.7 represents hydrogen, unsubstituted C.sub.1-4
alkyl or --C(.dbd.O)CH.sub.3, (b) R represents a group of formula
(Y1) as defined in claim 8; R.sup.8 is a group --COOH or an ester
group of formula --(C.dbd.O)OR.sup.14 wherein R.sup.14 is
R.sup.15R.sup.16R.sup.17C-- wherein either R.sup.15 is hydrogen or
C.sub.1-3alkyl-(Z.sup.1).sub.a--[(C.sub.1-C.sub.3)alkyl].sub.b-
wherein a and b are independently 0 or 1 and Z.sup.1 is --O--,
--S--, or --NH-- and R.sup.16 and R.sup.17 are independently
hydrogen or C.sub.1-3 alkyl-; or R.sup.15 and R.sup.16 taken
together with the carbon to which they are attached form a 3- to
7-membered cycloalkyl ring and R.sup.17 is hydrogen; R.sup.11
represents a hydrogen atom or an unsubstituted C.sub.1-2 alkyl
group; [Linker] is --(CH.sub.2).sub.m--X.sup.1-(Alk.sup.1).sub.x-
or --(CH.sub.2).sub.v--, wherein v is 1 or 2, m is 0, 1, 2 or 3; x
is 0 or 1; Alk.sup.1 is an unsubstituted C.sub.1-3 alkylene group;
and X.sup.1 is a bond, --O--, --S--, --NR.sup.7--, --C(.dbd.O)-- or
--C(.dbd.O)NR.sup.5--, wherein R.sup.5 is hydrogen or C.sub.1-4
alkyl and R.sup.7 represents hydrogen, unsubstituted C.sub.1-4
alkyl or --C(.dbd.O)CH.sub.3, wherein when x is 0, X.sup.1 is a
bond or C(.dbd.O), (c) R represents a group of formula (Y2) as
defined in claim 9; R.sup.8 is a group --COOH or an ester group of
formula --(C.dbd.O)OR.sup.14 wherein R.sup.14 is
R.sup.15R.sup.16R.sup.17C-- wherein either R.sup.15 is hydrogen or
C.sub.1-3alkyl-(Z.sup.1).sub.a--[(C.sub.1-C.sub.3)alkyl].sub.b-
wherein a and b are independently 0 or 1 and Z.sup.1 is --O--,
--S--, or --NH-- and R.sup.16 and R.sup.17 are independently
hydrogen or C.sub.1-3 alkyl-; or R.sup.15 and R.sup.16 taken
together with the carbon to which they are attached form a 3- to
7-membered cycloalkyl ring and R.sup.17 is hydrogen; R.sup.11
represents a hydrogen atom or an unsubstituted C.sub.1-2 alkyl
group; [Linker] represents
--(CH.sub.2).sub.m--X.sup.1-(Alk.sup.1)-Y.sup.1, wherein m is 0, 1,
2 or 3; x is 0 or 1; X.sup.1 is a bond or --C(.dbd.O); Alk.sup.1 is
an unsubstituted C.sub.1-3 alkylene group; and Y.sup.1 is a bond or
--NR.sup.7--, preferably --NR.sup.7--, wherein R.sup.7 represents
hydrogen, unsubstituted C.sub.1-4 alkyl or --C(.dbd.O)CH.sub.3.
15. A compound as claimed in any one of claims 1 to 3 and 5 to 14,
wherein: ring A represents a phenyl group, said ring A group being
unsubstituted or substituted by 1, 2 or 3 substituents which are
the same or different and are selected from halogen atoms and
unsubstituted C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl and
--NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4 are the same
or different and represent hydrogen or unsubstituted C.sub.1-2
alkyl; R.sup.1 and R.sup.2 each represent hydrogen; n is 1; X is
NH; V is --N.dbd.; W is --C(H).dbd.; and ring B represents a
1,4-phenylene, 1,3-phenylene group or 1,3-pyrazolyl group, said
ring B group being unsubstituted or substituted by 1, 2 or 3
substituents which are the same or different and are selected from
halogen atoms and unsubstituted C.sub.1-4 alkyl, C.sub.1-4 alkoxy,
hydroxyl and --NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4
are the same or different and represent hydrogen or unsubstituted
C.sub.1-2 alkyl.
16. A compound as claimed in claim 15, wherein: R represents a
group of formula (Y1'): ##STR00140## R.sup.8 is a group --COOH or
an ester group of formula --(C.dbd.O)OR.sup.14 wherein R.sup.14 is
R.sup.15R.sup.16R.sup.17C-- wherein either R.sup.15 is hydrogen or
C.sub.1-3alkyl-(Z).sub.a--[(C.sub.1-C.sub.3)alkyl].sub.b- wherein a
and b are independently 0 or 1 and Z.sup.1 is --O--, --S--, or
--NH-- and R.sup.16 and R.sup.17 are independently hydrogen or
C.sub.1-3 alkyl-; or R.sup.15 and R.sup.16 taken together with the
carbon to which they are attached form a 3- to 7-membered
cycloalkyl ring and R.sup.17 is hydrogen; [Linker] is
--(CH.sub.2).sub.v-- and --C(.dbd.O)--, wherein v is 1 or 2.
17. A compound as claimed in claim 15, wherein: R represents a
group of formula (Y2'): ##STR00141## R.sup.8 is a group --COOH or
an ester group of formula --(C.dbd.O)OR.sup.14 wherein R.sup.14 is
R.sup.15R.sup.16R.sup.17C-- wherein either R.sup.15 is hydrogen or
C.sub.1-3alkyl-(Z.sup.1).sub.a--[(C.sub.1-C.sub.3)alkyl].sub.b-
wherein a and b are independently 0 or 1 and Z.sup.1 is --O--,
--S--, or --NH-- and R.sup.16 and R.sup.17 are independently
hydrogen or C.sub.1-3 alkyl-; or R.sup.15 and R.sup.16 taken
together with the carbon to which they are attached form a 3- to
7-membered cycloalkyl ring and R.sup.17 is hydrogen; [Linker] is
--C(.dbd.O)NR.sup.7--, --CH.sub.2NR.sup.7-- or
--CH.sub.2C(.dbd.O)NR.sup.7--, wherein R.sup.7 represents hydrogen,
unsubstituted methyl, unsubstituted ethyl, unsubstituted isobutyl
or --C(.dbd.O)CH.sub.3.
18. A compound as claimed in claim 1, which is: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate; Cyclopentyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
piperazine-2-carboxylate; Cyclopentyl
(2S)-4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
piperazine-2-carboxylate; Cyclopentyl
(2R)-4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
piperazine-2-carboxylate; Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-chlorobe-
nzyl)amino]piperidine-2-carboxylate; Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(eth-
yl)amino]piperidine-2-carboxylate; Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(met-
hyl)amino]piperidine-2-carboxylate; tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(eth-
yl)amino]piperidine-2-carboxylate; Cyclopentyl
(2R)-4-[acetyl(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzy-
l)amino]piperidine-2-carboxylate; Cyclopentyl
(2R)-4-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piper-
azine-2-carboxylate; Cyclopentyl
1-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amino]cyc-
lobutanecarboxylate; Cyclopentyl
N-[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]--
2-methylalaninate; Cyclopentyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-ethylbenzyl)pi-
perazine-2-carboxylate; or Cyclopentyl
(2S)-4-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piper-
azine-2-carboxylate.
19. A compound as defined in any one of the preceding claims for
use in a method of treatment of the human or animal body.
20. A pharmaceutical composition which comprises a compound as
defined in any one of claims 1 to 18 and one or more
pharmaceutically acceptable carrier(s) and/or excipients.
21. A compound as defined in any one of claims 1 to 18 for use in
the treatment of a disease or disorder mediated by CSF-1R
kinase.
22. A compound as defined in any one of claims 1 to 18 for use in
the treatment of a cell proliferative disease, polyglutamine
disease, neurodegenerative disease, autoimmune disease, diabetes,
haematological disease, inflammatory disease, cardiovascular
disease, atherosclerosis, or the inflammatory sequelia of
infection.
23. A compound for use according to claim 22 for use in the
treatment of cancer, rheumatoid arthritis, psoriatic arthritis,
Type 1 diabetes, asthma, inflammatory bowel disease, systemic lupus
erythematosis, inflammation accompanying infectious conditions,
psoriasis, Crohns disease, ulcerative colitis, chronic obstructive
pulmonary disease, multiple sclerosis, atopic dermatitis or graft
versus host disease.
24. A method of treating or preventing a disease or disorder
mediated by CSF-1R kinase in a subject, which method comprises
administering to said subject an effective amount of a compound as
defined in any one of claims 1 to 16.
25. Use of a compound as defined in any one of claims 1 to 18 in
the manufacture of a medicament for use in the treatment or
prevention of a disease or disorder mediated by CSF-1R kinase.
Description
[0001] The invention relates to a series of amino acid ester and
cyclic amino acid ester compounds, to compositions containing them,
to processes for their preparation and to their use in medicine as
inhibitors of CSF-1R for the treatment of diseases and disorders
mediated by CSF-1R kinase. Such diseases and disorders include cell
proliferative disease such as cancer and psoriasis, polyglutamine
disease such as Huntingdon's disease, neurodegenerative disease
such as Alzheimers disease, autoimmune disease such as rheumatoid
arthritis, diabetes, haematological disease, inflammatory disease,
cardiovascular disease, atherosclerosis, and the inflammatory
sequelia of infection.
BACKGROUND OF THE INVENTION
[0002] Colony-stimulating factor-1 receptor (FMS, also known as
CSF-1R) is a homodimeric, class III receptor tyrosine kinase for
colony-stimulating factor-1 (CSF-1 or macrophage colony-stimulating
factor, M-CSF) that is encoded by the FMS proto-oncogene and has
been the focus of recent reviews [Nat. Rev immunol. 2008, 8, 533.].
CSF-1 regulates the survival, proliferation, differentiation, and
function of the macrophage lineage, including monocytes, tissue
macrophages, dendritic cells, microglia, and osteoclasts [Trends
Cell Biol. 2004, 14, 628-638.]. These mononuclear phagocytes act as
a defense against invading pathogens by maintaining tissue
homeostasis. CSF-1 exists in 3 major isoforms, namely, a secreted
glycoprotein and a secreted proteoglycan, both of which circulate
throughout the body, and a cell surface protein, which is involved
in local regulation of target cells.
[0003] The class III receptor tyrosine kinases (RTKs), which
include c-FMS, c-Kit, Flt-3, and PDGFR, are 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.
[0004] Binding of CSF-1 to the extracellular domain of FMS
stabilizes receptor dimerization, induces trans-autophosphorylation
of the intracellular FMS domain, and activates downstream
cytoplasmic signaling, which leads to differentiation and
activation of macrophage lineage cells.
[0005] Small molecule inhibitors of the FMS active site block
receptor autophosphorylation and subsequently block the signals
that control the survival, expression, proliferation, and
differentiation of macrophages. Evidence linking macrophage numbers
to several diseases, including cancer and inflammatory disease, has
led to extensive efforts in developing small molecule inhibitors of
FMS. In studies, high macrophage levels in target tissues have been
correlated with disease severity in RA [Ann. Rheum. Dis. 2005, 64,
834-838] and immune nephritis [Nephrol. Dial. Transplant. 2001, 16,
1638-1647] and poor survivability and tumor progression [J. Clin.
Oncol. 2005, 23, 953-964. Cancer Res. 2006, 66, 605-612] in some
cancers. Consequently, regulation of macrophage numbers may be the
key role of CSF-1 function.
[0006] The colony-stimulating factors (CSFs) are a group of
hematopoietic cell growth factors that include M-CSF and
granulocyte colony-stimulating factor (G-CSF), both of which are
relatively lineage-specific, and granulocyte/macrophage CSF
(GM-CSF), which functions at earlier stages of cell lineage by
regulating hematopoietic progenitors [1]. Recently, a new ligand
for FMS has been identified, interleukin-34 (IL-34), which
functions as a specific and independent ligand of FMS, stimulates
FMS dependent phosphorylation of extracellular signal-regulated
kinase-1 and -2 (ERK1/2), and promotes the formation of the
colony-forming unit-macrophage (CFU-M) in human bone marrow
cultures [Science 2008, 320, 807-811]. FMS is a critical player in
the CSF-1 pathway and has emerged as a viable target for regulating
macrophage levels in multiple diseases including cancer,
inflammation, and autoimmunity. Therefore, the identification of
novel and selective small molecule CSF-1 antagonists has become the
focus of extensive competitive research efforts By regulating the
development and activation of mononuclear phagocytes, CSF-1 plays a
key role in the innate immune response to viral, bacterial, and
fungal infections and increases the efficiency of vaccination.
CSF-1 is also involved in promoting and sustaining inflammation in
several diseases. Early evidence that CSF-1 depletion has a
therapeutic benefit in autoimmune and inflammatory diseases was
observed in the collagen-induced arthritis (CIA) mouse model.
CSF-1-deficient mice were resistant to the development of arthritis
despite a normal immune response to type II collagen [23]. Indeed,
increased levels of CSF-1, like that found in the synovial fluid of
RA patients, demonstrate a strong correlation with disease severity
in conditions such as RA and immune nephritis. The proinflammatory
cytokines interleukin-1.beta. (IL-1.beta.) and TNF-.alpha. that
stimulate the production of these synovial macrophages also play a
contributing role. In the CIA model, both monoclonal antibodies and
small-molecule inhibitors of CSF-1 have shown efficacy in
decreasing macrophage numbers in joints and slowing the overall
progression of disease.
BRIEF DESCRIPTION OF THE INVENTION
[0007] The present invention makes available a new class of
inhibitors of CSF-1R having pharmaceutical utility in the treatment
of diseases and disorders mediated by CSF-1R kinase such as cancer,
autoimmune and inflammatory diseases. The inhibitors of the
invention are characterised by having an alpha amino acid motif or
an alpha amino acid ester motif, which is a substrate for
intracellular carboxyesterase (also referred to herein as an
"esterase motif") covalently linked to the parent molecular
template. The hydrophobic esterase motif allows the molecule to
pass through the cell wall, and thereby allows intracellular
carboxyesterase activity to hydrolyse the ester to release the
parent acid. Being charged, the acid is not readily transported out
of the cell, where it therefore accumulates to increase the
intracellular concentration of active CSF-1R inhibitor. This leads
to increases in potency and duration of action relative to the
parent CSF-1R inhibitor template.
[0008] Accordingly, the present invention provides a compound which
is an amino acid or amino acid ester of formula (I) or a salt,
N-oxide, hydrate or solvate thereof:
##STR00002##
wherein: ring A is a C.sub.6-12 aryl, 5- to 12-membered
heterocyclyl or C.sub.3-7 carbocyclyl ring; R.sup.1 and R.sup.2
independently represent hydrogen, halogen or an unsubstituted group
selected from C.sub.1-4 alkyl, C.sub.2-4 alkenyl, C.sub.1-4 alkoxy,
C.sub.2-4 alkenyloxy, C.sub.1-4 haloalkyl, C.sub.2-4 haloalkenyl,
C.sub.1-4 haloalkoxy, C.sub.2-4 haloalkenyloxy, hydroxyl,
--SR.sup.3, cyano, nitro, C.sub.1-4 hydroxyalkyl and
--NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4 are the same
or different and represent hydrogen or unsubstituted C.sub.1-2
alkyl; n is 0 or 1;
X is NH or O;
[0009] V and W independently represent --N.dbd. or --C(Z).dbd.;
each Z is the same or different and represents hydrogen, fluoro,
chloro, bromo, iodo, cyano, unsubstituted C.sub.1-3 fluoroalkyl or
unsubstituted C.sub.1-3alkyl; ring B is a C.sub.6-12 aryl, 5- to
12-membered heterocyclyl or C.sub.3-7 carbocyclyl ring; [Linker]
represents a group of formula
--(CH.sub.2).sub.m--X.sup.1-L.sup.1-Y.sup.1-- wherein: [0010] m is
0, 1, 2 or 3; [0011] X.sup.1 represents a bond, --O--, --S--,
--NR.sup.7--, --C(.dbd.O)--, --C(.dbd.O)O--, --OC(.dbd.O)--,
--S(.dbd.O).sub.2--; --NR.sup.5C(.dbd.O)--, --C(.dbd.O)NR.sup.5--,
--NR.sup.5C(.dbd.O)NR.sup.6--, --NR.sup.5S(.dbd.O).sub.2--, or
--S(.dbd.O).sub.2NR.sup.5-- wherein R.sup.5 and R.sup.6 are
independently hydrogen or C.sub.1-6 alkyl and R.sup.7 represents
hydrogen, unsubstituted C.sub.1-4 alkyl or --C(.dbd.O)CH.sub.3;
[0012] L.sup.1 represents a divalent radical of formula
-(Alk.sup.1).sub.x-(Q).sub.y-(Het).sub.w-(Alk.sup.2).sub.z- or
-(Alk.sup.1).sub.x-(Het).sub.w-(Q).sub.y-(Alk.sup.2).sub.z- wherein
[0013] x, y, w and z are independently 0 or 1; [0014] Q represents
a divalent C.sub.6-12 aryl, 5- to 12-membered heterocyclyl or
C.sub.3-7 carbocyclyl ring; [0015] Het represents --O--, --S-- or
--NR.sup.7-- wherein R.sup.7 represents hydrogen, unsubstituted
C.sub.1-4 alkyl or --C(.dbd.O)CH.sub.3; [0016] Alk.sup.1 and
Alk.sup.2 independently represent divalent C.sub.3-7 cycloalkyl
radicals, or straight or branched, C.sub.1-6 alkylene, C.sub.2-6
alkenylene, or C.sub.2-6 alkynylene radicals; [0017] Y.sup.1
represents a bond, --O--, --S--, --NR.sup.7--, --C(.dbd.O)--,
--C(.dbd.O)O--, --OC(.dbd.O)--, --S(.dbd.O).sub.2--,
--NR.sup.5C(.dbd.O)--, --C(.dbd.O)NR.sup.5--,
--NR.sup.5C(.dbd.O)NR.sup.6--, --NR.sup.5S(.dbd.O).sub.2--, or
--S(.dbd.O).sub.2NR.sup.5--, wherein R.sup.5 and R.sup.6 are
independently hydrogen or C.sub.1-6 alkyl and R.sup.7 represents
hydrogen, unsubstituted C.sub.1-4 alkyl or --C(.dbd.O)CH.sub.3;
[0018] R represents a group of formula (X) or (Y):
[0018] ##STR00003## [0019] in which: [0020] R.sup.8 is a group
--COOH or an ester group which is hydrolysable by one or more
intracellular carboxylesterase enzymes to a --COOH group; [0021]
R.sup.9 and R.sup.10 are the same or different and each represents
the .alpha.-substituent of a natural or non-natural .alpha.-amino
acid, or R.sup.9 and R.sup.10, taken together with the carbon to
which they are attached, form a 3- to 6-membered saturated spiro
cycloalkyl or heterocyclyl ring; [0022] R.sup.11 represents a
hydrogen atom or a C.sub.1-4 alkyl group; [0023] ring D is a 5- to
7-membered saturated heterocyclyl group having at least one
nitrogen atom in the ring, wherein R.sup.8 and R.sup.11 are linked
to a ring carbon adjacent to a ring nitrogen, and wherein the bond
shown intersected by a wavy line may be from a nitrogen atom or a
carbon atom in ring D; wherein when R is (X), [Linker] is not
connected to (X) via an O, N or S atom; and wherein, unless
otherwise stated: [0024] any alkyl, alkenyl and alkynyl groups and
moieties in R.sup.5, R.sup.6, R.sup.8, R.sup.9, R.sup.10, R.sup.11,
Alk.sup.1 and Alk.sup.2 are the same or different and are each
unsubstituted or substituted with 1, 2 or 3 unsubstituted
substituents which are the same or different and are selected from
halogen atoms and C.sub.1-4 alkoxy, C.sub.2-4 alkenyloxy, C.sub.1-4
haloalkoxy, C.sub.2-4 haloalkenyloxy, hydroxyl, --SR.sup.3, cyano,
nitro and --NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4 are
the same or different and represent hydrogen or unsubstituted
C.sub.1-2 alkyl; [0025] any aryl, heterocyclyl, cycloalkyl and
carbocyclyl groups and moieties in rings A, B, Q, D, Alk.sup.1 and
Alk.sup.2 and the ring formed by R.sup.9 and R.sup.10 are the same
or different and are each unsubstituted or substituted by 1, 2, 3
or 4 unsubstituted substituents selected from halogen atoms, and
cyano, nitro, C.sub.1-4 alkyl, C.sub.1-4 alkoxy, C.sub.2-4 alkenyl,
C.sub.2-4 alkenyloxy, C.sub.1-4 haloalkyl, C.sub.2-4 haloalkenyl,
C.sub.1-4 haloalkoxy, C.sub.2-4 haloalkenyloxy, hydroxyl, C.sub.1-4
hydroxyalkyl, --SR.sup.3 and --NR.sup.3R.sup.4 groups wherein each
R.sup.3 and R.sup.4 is the same or different and represents
hydrogen or unsubstituted C.sub.1-2 alkyl, or from substituents of
formula --COOR.sup.12, --COR.sup.12, --SO.sub.2R.sup.12,
--CONR.sup.12R.sup.13, --SO.sub.2NR.sup.12R.sup.13,
--OCONR.sup.12R.sup.13, --NR.sup.12COR.sup.13,
--NR.sup.12COOR.sup.13, --NR.sup.12SO.sub.2R.sup.13,
--NR.sup.12SO.sub.2OR.sup.13 or --NR.sup.12CONR.sup.12R.sup.13
wherein R.sup.12 and R.sup.13 are the same or different and
represent hydrogen or unsubstituted C.sub.1-4 alkyl, or R.sup.12
and R.sup.13 when attached to the same nitrogen atom form a
non-fused 5- or 6-membered heterocyclyl group, with the proviso
that the compound is not:
4-(4-{6-Amino-5-[1-(2,6-dichloro-3-fluoro-phenyl)-ethoxy]-pyridin-3-yl}-p-
yrazol-1-yl)-pyrrolidine-2-carboxylic acid methyl ester. It is
preferable, in line with general principles of medicinal chemistry
practice, that the compounds with which this invention is concerned
should have molecular weights of no more than 600.
[0026] The present invention also provides a compound as defined
above for use in a method of treatment of the human or animal
body.
[0027] The present invention further provides a pharmaceutical
composition which comprises a compound as defined above and one or
more pharmaceutically acceptable carrier(s) and/or excipients.
[0028] In another aspect, the present invention provides a compound
as defined above for use in the treatment of a disease or disorder
mediated by CSF-1R kinase. The invention also provides use of a
compound as defined above in the manufacture of a medicament for
use in the treatment or prevention of a disease or disorder
mediated by CSF-1R kinase.
[0029] Still further, the invention provides a method of treating
or preventing a disease or disorder mediated by CSF-1R kinase in a
subject, which method comprises administering to said subject an
effective amount of a compound as defined above.
DETAILED DESCRIPTION OF THE INVENTION
Terminology
[0030] The alkyl, alkenyl and alkynyl groups and moieties in
R.sup.5, R.sup.6, R.sup.8, R.sup.9, R.sup.10, R.sup.11, Alk.sup.1,
and Alk.sup.2 are unsubstituted or substituted with 1, 2 or 3,
preferably 1 or 2, unsubstituted substituents which are the same or
different and are selected from halogen atoms and C.sub.1-4 alkoxy,
C.sub.2-4 alkenyloxy, C.sub.1-4 haloalkoxy, C.sub.2-4
haloalkenyloxy, hydroxyl, --SR.sup.3, cyano, nitro and
--NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4 are the same
or different and represent hydrogen or unsubstituted C.sub.1-2
alkyl. More preferred substituents are halogen, C.sub.1-4 alkoxy,
hydroxyl and --NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4
are the same or different and represent hydrogen or unsubstituted
C.sub.1-2 alkyl.
[0031] When the alkyl, alkylene, alkenylene and alkynylene moieties
above are substituted by two or three substituents, it is preferred
that not more than two substituents are selected from hydroxyl,
cyano and nitro. More preferably, not more than one substituent is
selected from hydroxyl, cyano and nitro.
[0032] As used herein, a C.sub.1-6 alkyl group or moiety is a
linear or branched alkyl group or moiety containing from 1 to 6
carbon atoms, for example a C.sub.1-4 alkyl group or moiety
containing from 1 to 4 carbon atoms. Examples of C.sub.1-4 alkyl
groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl
and t-butyl. For the avoidance of doubt, where two alkyl moieties
are present in a group, the alkyl moieties may be the same or
different.
[0033] As used herein, a C.sub.2-6 alkenyl group or moiety is a
linear or branched alkenyl group or moiety having at least one
double bond of either E or Z stereochemistry where applicable and
containing from 2 to 6 carbon atoms, for example a C.sub.2-4
alkenyl group or moiety containing from 2 to 4 carbon atoms, such
as --CH.dbd.CH.sub.2 or --CH.sub.2--CH.dbd.CH.sub.2,
--CH.sub.2--CH.sub.2--CH.dbd.CH.sub.2,
--CH.sub.2--CH.dbd.CH--CH.sub.3, --CH.dbd.C(CH.sub.3)--CH.sub.3 and
--CH.sub.2--C(CH.sub.3).dbd.CH.sub.2. For the avoidance of doubt,
where two alkenyl moieties are present in a group, they may be the
same or different.
[0034] As used herein, a C.sub.1-6 alkylene group or moiety is a
linear or branched alkylene group or moiety, for example a
C.sub.1-4 alkylene group or moiety. Examples include methylene,
n-ethylene, n-propylene and --C(CH.sub.3).sub.2-- groups and
moieties.
[0035] As used herein, a C.sub.2-6 alkenylene group or moiety is a
linear or branched alkenylene group or moiety, for example a
C.sub.2-4 alkenylene group or moiety. Examples include
--CH.dbd.CH--, --CH.dbd.CH--CH.sub.2--, --CH.sub.2--CH.dbd.CH-- and
--CH.dbd.CH--CH.dbd.CH--.
[0036] As used herein, a C.sub.2-6 alkynylene group or moiety is a
linear or branched alkynylene group or moiety, for example a
C.sub.2-4 alkynylene group or moiety. Examples include
--C.ident.C--, --C.ident.C--CH.sub.2-- and
--CH.sub.2--C.ident.C--.
[0037] As used herein, a halogen atom is chlorine, fluorine,
bromine or iodine.
[0038] As used herein, a C.sub.1-4 alkoxy group or C.sub.2-4
alkenyloxy group is typically a said C.sub.1-4 alkyl group or a
said C.sub.2-4 alkenyl group respectively which is attached to an
oxygen atom.
[0039] A haloalkyl, haloalkenyl, haloalkoxy or haloalkenyloxy group
is typically a said alkyl, alkenyl, alkoxy or alkenyloxy group
respectively which is substituted by one or more said halogen
atoms. Typically, it is substituted by 1, 2 or 3 said halogen
atoms.
[0040] Preferred haloalkyl and haloalkoxy groups include
perhaloalkyl and perhaloalkoxy groups such as --CX.sub.3 and
--OCX.sub.3 wherein X is a said halogen atom, for example chlorine
or fluorine. A fluoroalkyl group is typically a said alkyl group
which is substituted by one or more fluorine atoms. Typically, it
is substituted by 1, 2 or 3 said fluorine atoms.
[0041] As used herein, a C.sub.1-4 hydroxyalkyl group is a
C.sub.1-4 alkyl group substituted by one or more hydroxy groups.
Typically, it is substituted by one, two or three hydroxy groups.
Preferably, it is substituted by a single hydroxy group.
[0042] As used herein, a C.sub.6-12 aryl group or moiety is a
monocyclic, 6- to 12-membered aromatic hydrocarbon ring having from
6 to 12 carbon atoms, for example phenyl or naphtyl. Phenyl is
preferred.
[0043] As used herein the unqualified term "heteroaryl" refers to a
mono-, bi- or tri-cyclic aromatic radical containing one or more,
for example 1, 3 or 4, in particular 1 or 2, heteroatoms selected
from S, N and O, and includes radicals having two such monocyclic
rings, or one such monocyclic ring and one monocyclic aryl ring,
which are directly linked by a covalent bond. Illustrative of such
radicals are thienyl, benzthienyl, furyl, benzfuryl, pyrrolyl,
imidazolyl, benzimidazolyl, thiazolyl, benzthiazolyl, isothiazolyl,
benzisothiazolyl, pyrazolyl, oxazolyl, benzoxazolyl, isoxazolyl,
benzisoxazolyl, isothiazolyl, triazolyl, benztriazolyl,
thiadiazolyl, oxadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl,
pyrazinyl, triazinyl, indolyl and indazolyl, in particular
pyrazole.
[0044] As used herein the unqualified term "heterocyclyl" or
"heterocyclic" includes "heteroaryl" as defined above, and in its
non-aromatic meaning relates to a saturated or unsaturated mono-,
bi- or tri-cyclic non-aromatic radical containing one or more, for
example 1, 2, 3 or 4 in particular 1 or 2, heteroatoms selected
from S, N and O, and to groups consisting of a monocyclic
non-aromatic radical containing one or more such heteroatoms which
is covalently linked to another such radical or to a monocyclic
carbocyclic radical. Illustrative of such radicals are pyrrolyl,
furanyl, thienyl, piperidinyl, imidazolyl, oxazolyl, isoxazolyl,
thiazolyl, thiadiazolyl, pyrazolyl, pyridinyl, pyrrolidinyl,
pyridazinyl, pyrimidinyl, pyrazinyl, morpholinyl, piperazinyl,
indolyl, benzfuranyl, pyranyl, isoxazolyl, benzimidazolyl,
methylenedioxyphenyl, ethylenedioxyphenyl, maleimido and
succinimido groups, in particular piperidinyl, piperazinyl and
pyrazolyl.
[0045] As used herein, a C.sub.3-7 carbocyclic group or moiety is a
non-aromatic saturated or unsaturated hydrocarbon ring having from
3 to 7 carbon atoms. Preferably it is a saturated or
mono-unsaturated hydrocarbon ring (i.e. a cycloalkyl moiety or a
cycloalkenyl moiety) having from 3 to 7 carbon atoms, more
preferably having from 3 to 6 carbon atoms. Examples include
cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl and their
mono-unsaturated variants. Preferred examples of C.sub.3-7
carbocyclic group or moieties are cyclopentyl and cyclohexyl. A
C.sub.3-7 carbocyclyl group or moiety also includes C.sub.3-7
carbocyclyl groups or moieties described above but wherein one or
more ring carbon atoms are replaced by a group --C(O)--. More
preferably, 0, 1 or 2 ring carbon atoms (most preferably 0 or 2)
are replaced by --C(O)--. A preferred such group is
benzoquinone.
[0046] As used herein the term "cycloalkyl" refers to a monocyclic
saturated carbocyclic radical having from 3-8 carbon atoms and
includes, for example, cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl and cyclooctyl.
[0047] When the aryl, heterocyclyl, cycloalkyl and carbocyclyl
groups and moieties in rings A, B, Q and D and the rings formed by
Alk.sup.1 and Alk.sup.2 and by R.sup.9 and R.sup.10 are substituted
by two, three or four substituents, it is preferred that not more
than two substituents are selected from hydroxyl, cyano and nitro.
More preferably, not more than one substituent is selected from
hydroxyl, cyano and nitro. Furthermore, when the aryl, heteroaryl,
heterocyclyl and carbocyclyl moieties are substituted by two or
three substituents, it is preferred that not more than one
substituent is selected from --COOR.sup.12, --COR.sup.12,
--SO.sub.2R.sup.12, --CONR.sup.12R.sup.13,
--SO.sub.2NR.sup.12R.sup.13, --OCONR.sup.12R.sup.13,
--NR.sup.12COR.sup.13, --NR.sup.12COOR.sup.13,
--NR.sup.12SO.sub.2R.sup.13, --NR.sup.12SO.sub.2OR.sup.13 or
--NR.sup.12CONR.sup.12R.sup.13
[0048] As used herein the term "salt" includes base addition, acid
addition and quaternary salts. Compounds of the invention which are
acidic can form salts, including pharmaceutically acceptable salts,
with bases such as alkali metal hydroxides, e.g. sodium and
potassium hydroxides; alkaline earth metal hydroxides e.g. calcium,
barium and magnesium hydroxides; with organic bases e.g.
N-methyl-D-glucamine, choline tris(hydroxymethyl)amino-methane,
L-arginine, L-lysine, N-ethyl piperidine, dibenzylamine and the
like. Those compounds which are basic can form salts, including
pharmaceutically acceptable salts with inorganic acids, e.g. with
hydrohalic acids such as hydrochloric or hydrobromic acids,
sulphuric acid, nitric acid or phosphoric acid and the like, and
with organic acids e.g. with acetic, tartaric, succinic, fumaric,
maleic, malic, salicylic, citric, methanesulphonic,
p-toluenesulphonic, benzoic, benzenesulfonic, glutamic, lactic, and
mandelic acids and the like.
[0049] It is expected that compounds of the invention may be
recovered in hydrate or solvate form. The term `solvate` is used
herein to describe a molecular complex comprising the compound of
the invention and a stoichiometric amount of one or more
pharmaceutically acceptable solvent molecules, for example,
ethanol. The term `hydrate` is employed when said solvent is
water.
[0050] Compounds of the invention which contain one or more actual
or potential chiral centres, because of the presence of asymmetric
carbon atoms, can exist as a number of diastereoisomers with R or S
stereochemistry at each chiral centre. The invention includes all
such diastereoisomers and mixtures thereof.
[0051] Preferably ring A represents phenyl or a 5- to 6-membered
heterocyclyl group, said ring A group being unsubstituted or
substituted by 1, 2 or 3 substituents which are the same or
different and are selected from halogen atoms and unsubstituted
C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, C.sub.1-4 haloalkyl,
C.sub.1-4 haloalkoxy, C.sub.1-4 hydroxyalkyl, cyano, nitro,
--SR.sup.3 and --NR.sup.3R.sup.4 groups, wherein R.sup.3 and
R.sup.4 are the same or different and represent hydrogen or
unsubstituted C.sub.1-2 alkyl. More preferably, ring A represents a
phenyl group, said ring A group being unsubstituted or substituted
by 1, 2 or 3 substituents which are the same or different and are
selected from halogen atoms and unsubstituted C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, hydroxyl and --NR.sup.3R.sup.4 groups, wherein
R.sup.3 and R.sup.4 are the same or different and represent
hydrogen or unsubstituted C.sub.1-2 alkyl. For example, ring A may
represent a phenyl group, said ring A group being unsubstituted or
substituted by 1, 2 or 3 substituents which are the same or
different and are selected from fluorine and chlorine.
[0052] Preferably R.sup.1 and R.sup.2 independently represent
hydrogen, halogen or unsubstituted C.sub.1-4 alkyl; more preferably
hydrogen.
[0053] Preferably n is 1.
[0054] Preferably X is NH.
[0055] The ring containing V and W may contain a total of 1, 2 or 3
nitrogen atoms. In one embodiment, V is --N.dbd. and W is
--C(Z).dbd.. In another embodiment, V is --C(Z).dbd. and W is
--N.dbd.. Preferably, V is --N.dbd. and W is --C(Z).dbd.; more
preferably --CH.dbd..
[0056] Preferably Z represents hydrogen, fluoro, chloro, or
unsubstituted C.sub.1-3 alkyl; more preferably hydrogen.
[0057] Preferably ring B represents phenyl or a 5- to 6-membered
heterocyclyl group, preferably phenyl or a 5- to 6-membered
heterocyclyl group containing 1, 2 or 3 nitrogen atoms, for example
pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl or pyrazolyl, in
particular pyridinyl, pyridazinyl, pyrimidinyl or pyrazinyl, said
ring B group being unsubstituted or substituted by 1, 2 or 3
substituents which are the same or different and are selected from
halogen atoms and unsubstituted C.sub.1-4 alkyl, C.sub.1-4 alkoxy,
hydroxyl, C.sub.1-4 haloalkyl, C.sub.1-4 haloalkoxy, C.sub.1-4
hydroxyalkyl, cyano, nitro, --SR.sup.3 and --NR.sup.3R.sup.4
groups, wherein R.sup.3 and R.sup.4 are the same or different and
represent hydrogen or unsubstituted C.sub.1-2 alkyl.
[0058] More preferably, ring B represents a 1,4-phenylene group,
1,3-phenylene group, 1,3-pyrazolyl group or pyridinyl group, in
particular a 1,4-phenylene group, 1,3-phenylene group or
1,3-pyrazolyl group, said ring B group being unsubstituted or
substituted by 1, 2 or 3 substituents which are the same or
different and are selected from halogen atoms and unsubstituted
C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl and --NR.sup.3R.sup.4
groups, wherein R.sup.3 and R.sup.4 are the same or different and
represent hydrogen or unsubstituted C.sub.1-2 alkyl. For example,
ring B may represent an unsubstituted 1,3-pyrazolyl group, or a
1,4-phenylene or 1,3-phenylene group which is unsubstituted or
substituted by 1, 2 or 3 substituents which are the same or
different and are selected from fluorine, chlorine, methyl and
ethyl.
[0059] Typically, ring B represents a 1,4-phenylene group,
1,3-phenylene group or pyridinyl group, in particular a
1,4-phenylene group or 1,3-phenylene group, said ring B group being
unsubstituted or substituted by 1, 2 or 3 substituents which are
the same or different and are selected from halogen atoms and
unsubstituted C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl and
--NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4 are the same
or different and represent hydrogen or unsubstituted C.sub.1-2
alkyl. For example, ring B may represent a 1,4-phenylene or
1,3-phenylene group which is unsubstituted or substituted by 1, 2
or 3 substituents which are the same or different and are selected
from fluorine, chlorine, methyl and ethyl.
[0060] When B is as defined hereinabove, X is preferably NH.
[0061] [Linker] represents a group of formula
--(CH.sub.2).sub.m--X.sup.1-L.sup.1-Y.sup.1--.
[0062] Preferably, m is 0, 1 or 2.
[0063] In some embodiments, X.sup.1 may preferably represent a
bond, --O--, --S--, --NR.sup.7--, --C(.dbd.O)-- or
--C(.dbd.O)NR.sup.5--, wherein R.sup.5 is hydrogen or C.sub.1-4
alkyl and R.sup.7 represents hydrogen, unsubstituted C.sub.1-4
alkyl or --C(.dbd.O)CH.sub.3. More preferably, X.sup.1 represents a
bond or --C(.dbd.O)--.
[0064] L.sup.1 represents a divalent radical of formula
-(Alk.sup.1).sub.x-(Q).sub.y-(Het).sub.w-(Alk.sup.2).sub.z- or
-(Alk.sup.1).sub.x-(Het).sub.w-(Q)-(Alk.sup.2).sub.z-.
[0065] Preferably Q represents phenyl or a 5- to 6-membered
heterocyclyl group which is unsubstituted or substituted with 1, 2
or 3 substituents which are the same or different and are selected
from halogen atoms and unsubstituted C.sub.1-4 alkyl, C.sub.1-4
alkoxy, hydroxyl and --NR.sup.3R.sup.4 groups, wherein R.sup.3 and
R.sup.4 are the same or different and represent hydrogen or
unsubstituted C.sub.1-2 alkyl. Preferably Q is unsubstituted
phenyl, preferably unsubstituted 1,4-phenylene.
[0066] Preferably Alk.sup.1 and Alk.sup.2 independently represent a
C.sub.1-3 alkylene, C.sub.2-3 alkenylene or C.sub.2-3 alkynylene
group which is unsubstituted or unsubstituted with 1, 2 or 3
unsubstituted substituents which are the same or different and are
selected from halogen atoms and C.sub.1-2 alkoxy, hydroxyl and
--NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4 are the same
or different and represent hydrogen or unsubstituted C.sub.1-2
alkyl. More preferably, Alk.sup.1 and Alk.sup.2 independently
represent unsubstituted C.sub.1-3 alkylene.
[0067] Preferably Het is --O-- or --NR.sup.7--, preferably wherein
R.sup.7 is hydrogen, unsubstituted methyl, unsubstituted ethyl,
unsubstituted isobutyl or --C(.dbd.O)CH.sub.3. More preferably, Het
is --O--.
[0068] In one embodiment, y and w are each 1 and at least one of x
and z is 1, for example L.sup.1 is -Alk.sup.1-Het-Q- or
-Q-Het-Alk.sup.2-. Preferably, Q represents phenyl or a 5- to
6-membered heterocyclyl group as described above, preferably
unsubstituted phenyl, preferably unsubstituted 1,4-phenylene;
Alk.sup.1 and Alk.sup.2 independently represent a C.sub.1-3
alkylene, C.sub.2-3 alkenylene or C.sub.2-3 alkynylene group as
described above, preferably unsubstituted C.sub.1-3 alkylene; and
Het represents --O-- or --NR.sup.7-- as described above, preferably
--O--.
[0069] In one preferred embodiment, at least one of x and z is 1, y
and w are each 0 and Alk.sup.1 and Alk.sup.2 independently
represent unsubstituted C.sub.1-3 alkylene. In another preferred
embodiment, x, y and z are each 0.
[0070] In some embodiments, Y.sup.1 may preferably represent a
bond, --O--, --S--, --NR.sup.7--, --C(.dbd.O)-- or
--C(.dbd.O)NR.sup.5--, wherein R.sup.5 and is hydrogen or C.sub.1-4
alkyl and R.sup.7 represents hydrogen, unsubstituted C.sub.1-4
alkyl or --C(.dbd.O)CH.sub.3. More preferably, Y.sup.1 represents a
bond or --NR.sup.7--, wherein R.sup.7 represents hydrogen,
unsubstituted C.sub.1-4 alkyl or --C(.dbd.O)CH.sub.3, preferably,
hydrogen, unsubstituted methyl, unsubstituted ethyl, unsubstituted
isobutyl or --C(.dbd.O)CH.sub.3.
[0071] In one embodiment, [Linker] represents
--(CH.sub.2).sub.m--X.sup.1-(Alk.sup.1).sub.x-Y.sup.1, wherein m,
x, X.sup.1, Y.sup.1 and Alk.sup.1 are as defined herein.
Preferably, Alk.sup.1 is an unsubstituted C.sub.1-3 alkylene group,
X.sup.1 and Y.sup.1 independently represent a bond, --O--, --S--,
--NR.sup.7--, --C(.dbd.O)-- or --C(.dbd.O)NR.sup.5--, wherein
R.sup.5 is hydrogen or C.sub.1-4 alkyl and R.sup.7 represents
hydrogen, unsubstituted C.sub.1-4 alkyl or --C(.dbd.O)CH.sub.3.
More preferably, X.sup.1 represents a bond or --C(.dbd.O)-- and
Y.sup.1 represents a bond or --NR.sup.7--, wherein R.sup.7
represents hydrogen, unsubstituted C.sub.1-4 alkyl or
--C(.dbd.O)CH.sub.3, preferably, hydrogen, unsubstituted methyl,
unsubstituted ethyl, unsubstituted isobutyl or --C(.dbd.O)CH.sub.3.
Most preferred [Linker] groups are --(CH.sub.2).sub.v--,
--C(.dbd.O)NR.sup.7--, --CH.sub.2NR.sup.7--,
--CH.sub.2C(.dbd.O)NR.sup.7-- and --C(.dbd.O)--, wherein v is 1 or
2 and R.sup.7 represents hydrogen, unsubstituted methyl,
unsubstituted ethyl, unsubstituted isobutyl or
--C(.dbd.O)CH.sub.3.
[0072] In one embodiment, R represents a group of formula (X). When
R is (X), [Linker] is not connected to (X) via an O, S or N atom of
the [Linker] moiety. Thus when R is (X) and x, y and z are each 0,
X.sup.1 and Y.sup.1 are preferably independently chosen from a
bond, --C(.dbd.O)--, --OC(.dbd.O)--, and --NR.sup.5C(.dbd.O)--,
wherein R.sup.5 is hydrogen or C.sub.1-6 alkyl; or when R is (X)
and at least one of x, y and z is 1, Y.sup.1 is preferably chosen
from a bond, --C(.dbd.O)--, --OC(.dbd.O)--, and
--NR.sup.5C(.dbd.O)--, and X.sup.1 is preferably chosen from a
bond, --O--, --S--, --NR.sup.7--, --C(.dbd.O)-- or
--C(.dbd.O)NR.sup.5--, wherein R.sup.5 is hydrogen or C.sub.1-4
alkyl and R.sup.7 represents hydrogen, unsubstituted C.sub.1-4
alkyl or --C(.dbd.O)CH.sub.3. Preferably, when R is (X), [Linker]
is --(CH.sub.2).sub.m--X.sup.1-(Alk.sup.1)- or
--(CH.sub.2).sub.v--, wherein v is 1 or 2, m is 0, 1, 2 or 3;
Alk.sup.1 is an unsubstituted C.sub.1-3 alkylene group and X.sup.1
is a bond, --O--, --S--, --NR.sup.7--, --C(.dbd.O)-- or
--C(.dbd.O)NR.sup.5--, wherein R.sup.5 is hydrogen or C.sub.1-4
alkyl and R.sup.7 represents hydrogen, unsubstituted C.sub.1-4
alkyl or --C(.dbd.O)CH.sub.3. Most preferred [Linker] groups when R
is (X) are groups --(CH.sub.2).sub.v-- wherein v is 1 or 2.
[0073] In another embodiment, R represents a group of formula
(Y1):
##STR00004##
wherein ring D is a 5- to 7-membered saturated heterocyclyl group
having at least two nitrogen atoms in the ring, wherein R.sup.8 and
R.sup.11 are linked to a ring carbon adjacent to a ring nitrogen,
and wherein the bond shown intersected by a wavy line is from a
nitrogen atom in ring D.
[0074] When R is (Y1), [Linker] is preferably not connected to (Y1)
via an O, S or N atom of the [Linker] moiety. Thus when R is (Y1)
and x, y and z are each 0, X.sup.1 and Y.sup.1 are preferably
chosen from a bond, --C(.dbd.O)--, --OC(.dbd.O)--, and
--NR.sup.5C(.dbd.O)--, wherein R.sup.5 is hydrogen or C.sub.1-6
alkyl, or when R is (Y1) and at least one of x, y and z is 1,
Y.sup.1 is preferably chosen from a bond, --C(.dbd.O)--,
--OC(.dbd.O)--, and --NR.sup.5C(.dbd.O)-- and X.sup.1 is preferably
chosen from a bond, --O--, --S--, --NR.sup.7--, --C(.dbd.O)-- or
--C(.dbd.O)NR.sup.5--, wherein R.sup.5 is hydrogen or C.sub.1-4
alkyl and R.sup.7 represents hydrogen, unsubstituted C.sub.1-4
alkyl or --C(.dbd.O)CH.sub.3. Preferably, when R is (Y1), [Linker]
is --(CH.sub.2).sub.m--X.sup.1-(Alk.sup.1).sub.x- or
--(CH.sub.2).sub.v--, wherein v is 1 or 2, m is 0, 1, 2 or 3; x is
0 or 1; Alk.sup.1 is an unsubstituted C.sub.1-3 alkylene group; and
X.sup.1 is a bond, --O--, --S--, --NR.sup.7--, --C(.dbd.O)-- or
--C(.dbd.O)NR.sup.5--, wherein R.sup.5 is hydrogen or C.sub.1-4
alkyl and R.sup.7 represents hydrogen, unsubstituted C.sub.1-4
alkyl or --C(.dbd.O)CH.sub.3, wherein when x is 0, X.sup.1 is a
bond or C(.dbd.O). Most preferred [Linker] groups are
--(CH.sub.2).sub.v-- and --C(.dbd.O)--, wherein v is 1 or 2.
[0075] Preferably the group of formula (Y1) is a group of formula
(Y1'):
##STR00005##
[0076] In another embodiment, R represents a group of formula
(Y2):
##STR00006##
wherein ring D is a 5- to 7-membered saturated heterocyclyl group
having at least one nitrogen atom in the ring, wherein R.sup.8 and
R.sup.11 are linked to a ring carbon adjacent to a ring nitrogen,
and wherein the bond shown intersected by a wavy line is from a
carbon atom in ring D.
[0077] Typically, when R is Y2, ring D is a 5- to 7-membered
saturated heterocyclyl group having at least one nitrogen atom in
the ring, wherein R.sup.8 and R.sup.11 are linked to a ring carbon
adjacent to a ring nitrogen, and wherein the bond shown intersected
by a wavy line is from a carbon atom in ring D, with the proviso
that D is not a pyrrolidinyl group. D may, for example, be a
pyrrolyl, piperidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl,
thiadiazolyl, pyrazolyl, pyridinyl, pyridazinyl, pyrimidinyl,
pyrazinyl, morpholinyl, piperazinyl group, for instance a
piperidinyl or piperazinyl group, for example a piperidinyl
group.
[0078] When R is (Y2), Y.sup.1 in [Linker] may preferably represent
a bond, --O--, --S--, --NR.sup.7--, --C(.dbd.O)-- or
--C(.dbd.O)NR.sup.5--, wherein R.sup.5 is hydrogen or C.sub.1-4
alkyl and R.sup.7 represents hydrogen, unsubstituted C.sub.1-4
alkyl or --C(.dbd.O)CH.sub.3. More preferably, Y.sup.1 represents a
bond or --NR.sup.7--, wherein R.sup.7 represents hydrogen,
unsubstituted C.sub.1-4 alkyl or --C(.dbd.O)CH.sub.3, preferably,
hydrogen, unsubstituted methyl, unsubstituted ethyl, unsubstituted
isobutyl or --C(.dbd.O)CH.sub.3. When R is (Y2), [Linker] is
preferably connected to (Y2) via an N atom of the [Linker] moiety.
Thus when R is (Y2), Y.sup.1 is preferably --NR.sup.7--, wherein
R.sup.7 represents hydrogen, unsubstituted C.sub.1-4 alkyl or
--C(.dbd.O)CH.sub.3, preferably, hydrogen, unsubstituted methyl,
unsubstituted ethyl, unsubstituted isobutyl or
--C(.dbd.O)CH.sub.3.
[0079] Thus, when R is (Y2), [Linker] typically represents
--(CH.sub.2).sub.m--X.sup.1-(Alk.sup.1).sub.x-Y.sup.1--, wherein m
is 0, 1, 2 or 3; x is 0 or 1; X.sup.1 is a bond, --O--, --S--,
--NR.sup.7--, --C(.dbd.O)-- or --C(.dbd.O)NR.sup.5--, preferably a
bond or --C(.dbd.O), wherein R.sup.5 is hydrogen or C.sub.1-4 alkyl
and R.sup.7 represents hydrogen, unsubstituted C.sub.1-4 alkyl or
--C(.dbd.O)CH.sub.3; Alk.sup.1 is an unsubstituted C.sub.1-3
alkylene group; and Y.sup.1 is a bond or --NR.sup.7--, preferably
--NR.sup.7--, wherein R.sup.7 represents hydrogen, unsubstituted
C.sub.1-4 alkyl or --C(.dbd.O)CH.sub.3. Most preferred [Linker]
groups are --C(.dbd.O)NR.sup.7--, --CH.sub.2NR.sup.7--,
--CH.sub.2CH.sub.2NR.sup.7--, NR.sup.7-- and
--CH.sub.2C(.dbd.O)NR.sup.7--, wherein R.sup.7 represents hydrogen,
unsubstituted methyl, unsubstituted ethyl, unsubstituted isobutyl
or --C(.dbd.O)CH.sub.3.
[0080] Preferably the group of formula (Y2) is a group of formula
(Y2'):
##STR00007##
[0081] Preferably in the above formulae
R.sup.8 is a group --COOH or an ester group of formula
--(C.dbd.O)OR.sup.14 wherein R.sup.14 is
R.sup.15R.sup.16R.sup.17C-- wherein [0082] (i) R.sup.15 is
hydrogen, fluorine or optionally substituted
C.sub.1-3alkyl-(Z.sup.1).sub.a--[(C.sub.1-C.sub.3)alkyl].sub.b-- or
C.sub.2-3alkenyl-(Z.sup.1).sub.a--[C.sub.1-3 alkyl].sub.b- wherein
a and b are independently 0 or 1 and Z.sup.1 is --O--, --S--, or
--NR.sup.18-- wherein R.sup.18 is hydrogen or C.sub.1-3 alkyl; and
R.sup.18 and R.sup.17 are independently hydrogen or C.sub.1-3
alkyl-; [0083] (ii) R.sup.15 is hydrogen or optionally substituted
R.sup.19R.sup.20N--C.sub.1-3alkyl--wherein R.sup.19 is hydrogen or
C.sub.1-3alkyl and R.sup.20 is hydrogen or C.sub.1-3alkyl; or
R.sup.19 and R.sup.20 together with the nitrogen to which they are
attached form an optionally substituted monocyclic heterocyclic
ring of 5- or 6-ring atoms or bicyclic heterocyclic ring system of
8 to 10 ring atoms, and R.sup.16 and R.sup.17 are independently
hydrogen or C.sub.1-3alkyl-; or [0084] (iii) R.sup.15 and R.sup.16
taken together with the carbon to which they are attached form an
optionally substituted monocyclic carbocyclic ring of from 3 to 7
ring atoms or bicyclic carbocyclic ring system of 8 to 10 ring
atoms, and R.sup.17 is hydrogen; or [0085] (iv) R.sup.15 and
R.sup.16 taken together with the carbon to which they are attached
form an optionally substituted monocyclic heterocyclic ring of from
3 to 7 ring atoms wherein at least one ring atom is selected from
--O--, --S--, or --NR.sup.8-- wherein R.sup.18 is hydrogen or
C.sub.1-3alkyl, or a bicyclic heterocyclic ring system of 8 to 10
ring atoms wherein at least one ring atom is selected from --O--,
--S--, or --NR.sup.18-- wherein R.sup.18 is hydrogen or
C.sub.1-3alkyl, and R.sup.17 is hydrogen.
[0086] Within these classes, R.sup.14 may be, for example methyl,
ethyl, n- or iso-propyl, n-, sec- or tert-butyl, dimethyl propyl,
pentanyl, cyclopentyl, methyl-substituted cyclopentyl,
cyclopentylmethyl, cyclohexyl, N-methylpiperidin-4-yl,
N-methylpyrrolidin-3-yl, tetrahydrofuran-3-yl, methoxyethyl,
indanyl, norbornyl, dimethylaminoethyl, or morpholinoethyl.
[0087] In one embodiment, either R.sup.15 is hydrogen or
C.sub.1-3alkyl-(Z.sup.1).sub.a--[(C.sub.1-C.sub.5)alkyl].sub.b-
wherein a and b are independently 0 or 1 and Z.sup.1 is --O--,
--S--, or --NH-- and R.sup.16 and R.sup.17 are independently
hydrogen or C.sub.1-3 alkyl-; or R.sup.15 and R.sup.16 taken
together with the carbon to which they are attached form a 3- to
7-membered cycloalkyl ring and R.sup.17 is hydrogen. Cyclopentyl
and t-butyl are particularly preferred at R.sup.14.
[0088] Preferably R.sup.9 and R.sup.10 are the same or different
and each represents the side chain of a natural amino acid or:
[0089] (i) a hydrogen atom; [0090] (ii) a C.sub.1-6 alkyl group;
[0091] (iii) R.sup.9 and R.sup.10, taken together with the carbon
to which they are attached, form a 3- to 6-membered saturated spiro
cycloalkyl ring; [0092] (iv) a group -L.sup.2-B.sup.1, in which
L.sup.2 represents a bond or a C.sub.1-6 alkylene group and B.sup.1
represents a C.sub.6-10 aryl or 5- to 10-membered heteroaryl group;
[0093] or [0094] (v) a group selected from indol-3-ylmethyl,
--CH.sub.2COOH, --CH.sub.2CH.sub.2COOH, --CH.sub.2CONH.sub.2,
--CH.sub.2CH.sub.2CONH.sub.2,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2NHC(NH)NH.sub.2, cyclohexyl,
cyclohexylmethyl and 1-benzylthio-1-methylethyl; [0095] said
C.sub.1-6 alkyl group in (ii) is unsubstituted or substituted with
1 or 2 substituents which are the same or different and represent
halogen, C.sub.1-4 alkoxy, C.sub.1-2 haloalkyl, hydroxyl,
--COOR.sup.3, --COONR.sup.3R.sup.4, --SR.sup.3 and
--NR.sup.3R.sup.4 wherein R.sup.3 and R.sup.4 are the same or
different and represent hydrogen or unsubstituted C.sub.1-2 alkyl;
[0096] said C.sub.1-6 alkylene group in (iv) is unsubstituted or
substituted with 1, 2 or 3 unsubstituted substituents which are the
same or different and are selected from halogen atoms and C.sub.1-4
alkoxy, hydroxyl, C.sub.1-2 haloalkyl and --NR.sup.3R.sup.4 groups
where R.sup.3 and R.sup.4 are the same or different and represent
hydrogen or unsubstituted C.sub.1-2 alkyl; and [0097] said
C.sub.6-10 aryl or 5- to 10-membered heteroaryl group in (iv) is
unsubstituted or substituted by 1, 2 or 3 substituents which are
the same or different and are selected from halogen atoms and
unsubstituted C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl,
C.sub.1-4 haloalkyl, C.sub.1-4 haloalkoxy, C.sub.1-4 hydroxyalkyl,
cyano, nitro, --SR.sup.3 and --NR.sup.3R.sup.4 groups where R.sup.3
and R.sup.4 are the same or different and represent hydrogen or
unsubstituted C.sub.1-2 alkyl.
[0098] More preferably, R.sup.9 and R.sup.10 independently
represent a hydrogen atom; a C.sub.1-6 alkyl group which is
unsubstituted or substituted with 1 or 2 substituents which are the
same or different and represent halogen, C.sub.1-4 alkoxy,
hydroxyl, --COOR.sup.3, --COONR.sup.3R.sup.4, --SR.sup.3 and
--NR.sup.3R.sup.4 wherein R.sup.3 and R.sup.4 are the same or
different and represent hydrogen or unsubstituted C.sub.1-2 alkyl;
R.sup.9 and R.sup.10, taken together with the carbon to which they
are attached, form a 3- to 6-membered saturated spiro cycloalkyl
ring; or a group of formula -L.sup.2-B.sup.1 wherein L.sup.2 is a
bond or an unsubstituted C.sub.1-4 alkylene group and B.sup.1
represents a phenyl or a 5- to 10-membered heteroaryl group which
is unsubstituted or substituted with one, two or three
unsubstituted substituents which are the same or different and
represent halogen atoms, C.sub.1-4 alkyl, C.sub.1-2 alkoxy,
C.sub.1-2 alkylthio and hydroxyl. Preferably, (i) R.sup.9 and
R.sup.10 are side chains of natural amino acids, (ii) one of
R.sup.9 and R.sup.10 is hydrogen or unsubstituted C.sub.1-4 alkyl
and the other is an unsubstituted C.sub.1-6 alkyl group or a
C.sub.1-6 alkyl group substituted with a C.sub.1-4 alkoxy group, or
(iii) R.sup.9 and R.sup.10, taken together with the carbon to which
they are attached, form a saturated spiro cyclobutyl ring. (ii) or
(iii) is preferred.
[0099] Preferably R.sup.11 represents a hydrogen atom or an
unsubstituted C.sub.1-2 alkyl group, preferably a hydrogen
atom.
[0100] In a preferred embodiment of the invention there is provided
a compound of formula (I) or a pharmaceutically acceptable salt,
N-oxide, hydrate or solvate thereof, in which: [0101] ring A
represents phenyl or a 5- to 6-membered heterocyclyl group, said
ring A group being unsubstituted or substituted by 1, 2 or 3
substituents which are the same or different and are selected from
halogen atoms and unsubstituted C.sub.1-4 alkyl, C.sub.1-4 alkoxy,
hydroxyl, C.sub.1-4 haloalkyl, C.sub.1-4 haloalkoxy, C.sub.1-4
hydroxyalkyl, cyano, nitro, --SR.sup.3 and --NR.sup.3R.sup.4
groups, wherein R.sup.3 and R.sup.4 are the same or different and
represent hydrogen or unsubstituted C.sub.1-2 alkyl; [0102] R.sup.1
and R.sup.2 each represent hydrogen; [0103] n is 1; [0104] X is NH;
[0105] V is --N.dbd.; [0106] W is --C(Z).dbd., wherein Z represents
hydrogen, fluoro, chloro, or unsubstituted C.sub.1-3 alkyl,
preferably hydrogen; and [0107] ring B represents phenyl or 5- to
6-membered heterocyclyl group, said ring B group being
unsubstituted or substituted by 1, 2 or 3 substituents which are
the same or different and are selected from halogen atoms and
unsubstituted C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl,
C.sub.1-4 haloalkyl, C.sub.1-4 haloalkoxy, C.sub.1-4 hydroxyalkyl,
cyano, nitro, --SR.sup.3 and --NR.sup.3R.sup.4 groups, wherein
R.sup.3 and R.sup.4 are the same or different and represent
hydrogen or unsubstituted C.sub.1-2 alkyl.
[0108] In this embodiment of the invention, the group [Linker]-R
preferably corresponds to one of (a) to (c) set out below:
(a) R represents a group of formula (X); [0109] R.sup.8 is as set
out below; [0110] R.sup.9 and R.sup.10 are side chains of natural
amino acids, one of R.sup.9 and R.sup.10 is hydrogen or
unsubstituted C.sub.1-4 alkyl and the other is an unsubstituted
C.sub.1-6 alkyl group or a C.sub.1-6 alkyl group substituted with a
C.sub.1-4 alkoxy group, or R.sup.9 and R.sup.10, taken together
with the carbon to which they are attached, form a saturated spiro
cyclobutyl ring; [0111] R.sup.11 represents a hydrogen atom or an
unsubstituted C.sub.1-2 alkyl group; [0112] [Linker] represents
--(CH.sub.2).sub.m--X.sup.1-(Alk.sup.1)- or --(CH.sub.2).sub.v--,
wherein v is 1 or 2, m is 0, 1, 2 or 3; Alk.sup.1 is an
unsubstituted C.sub.1-3 alkylene group and X.sup.1 is a bond,
--O--, --S--, --NR.sup.7--, --C(.dbd.O)-- or --C(.dbd.O)NR.sup.5--,
wherein R.sup.5 is hydrogen or C.sub.1-4 alkyl and R.sup.7
represents hydrogen, unsubstituted C.sub.1-4 alkyl or
--C(.dbd.O)CH.sub.3. (b) R represents a group of formula (Y1);
[0113] R.sup.8 is as set out below; [0114] R.sup.11 represents a
hydrogen atom or an unsubstituted C.sub.1-2 alkyl group; [0115]
[Linker] is --(CH.sub.2).sub.m--X.sup.1-(Alk.sup.1)- or
--(CH.sub.2).sub.v--, wherein v is 1 or 2, m is 0, 1, 2 or 3;
[0116] x is 0 or 1; Alk.sup.1 is an unsubstituted C.sub.1-3
alkylene group; and X.sup.1 is a bond, --O--, --S--, --NR.sup.7--,
--C(.dbd.O)-- or --C(.dbd.O)NR.sup.5--, wherein R.sup.5 is hydrogen
or C.sub.1-4 alkyl and R.sup.7 represents hydrogen, unsubstituted
C.sub.1-4 alkyl or --C(.dbd.O)CH.sub.3, wherein when x is 0,
X.sup.1 is a bond or C(.dbd.O). (c) R represents a group of formula
(Y2); [0117] R.sup.8 is as set out below; [0118] R.sup.11
represents a hydrogen atom or an unsubstituted C.sub.1-2 alkyl
group; [0119] [Linker] represents
--(CH.sub.2).sub.m--X.sup.1-(Alk.sup.1).sub.x-Y.sup.1, wherein m is
0, 1, 2 or 3; x is 0 or 1; [0120] X.sup.1 is a bond or --C(.dbd.O);
Alk.sup.1 is an unsubstituted C.sub.1-3 alkylene group; and Y.sup.1
is a bond or --NR.sup.7--, preferably --NR.sup.7--, wherein R.sup.7
represents hydrogen, unsubstituted C.sub.1-4 alkyl or
--C(.dbd.O)CH.sub.3.
[0121] Typically, in this embodiment, when R is Y2, ring D is a 5-
to 7-membered saturated heterocyclyl group having at least one
nitrogen atom in the ring, wherein R.sup.8 and R.sup.11 are linked
to a ring carbon adjacent to a ring nitrogen, and wherein the bond
shown intersected by a wavy line is from a carbon atom in ring D,
with the proviso that D is not a pyrrolidinyl group. D may, for
example, be a pyrrolyl, piperidinyl, imidazolyl, oxazolyl,
isoxazolyl, thiazolyl, thiadiazolyl, pyrazolyl, pyridinyl,
pyridazinyl, pyrimidinyl, pyrazinyl, morpholinyl, piperazinyl
group, for instance a piperidinyl or piperazinyl group, for example
a piperidinyl group.
[0122] In this embodiment, preferably, ring B represents a
1,4-phenylene group, 1,3-phenylene group or pyridinyl group, in
particular a 1,4-phenylene group or 1,3-phenylene group, said ring
B group being unsubstituted or substituted by 1, 2 or 3
substituents which are the same or different and are selected from
halogen atoms and unsubstituted C.sub.1-4 alkyl, C.sub.1-4 alkoxy,
hydroxyl and --NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4
are the same or different and represent hydrogen or unsubstituted
C.sub.1-2 alkyl. For example, ring B may represent a 1,4-phenylene
or 1,3-phenylene group which is unsubstituted or substituted by 1,
2 or 3 substituents which are the same or different and are
selected from fluorine, chlorine, methyl and ethyl.
[0123] In each of the above (a) to (c), R.sup.8 is a group --COOH
or an ester group of formula --(C.dbd.O)OR.sup.14 wherein R.sup.14
is R.sup.15R.sup.16R.sup.17C-- wherein either R.sup.15 is hydrogen
or C.sub.1-3alkyl-(Z.sup.1).sub.a--[(C.sub.1-C.sub.3)alkyl].sub.b-
wherein a and b are independently 0 or 1 and Z.sup.1 is --O--,
--S--, or --NH-- and R.sup.15 and R.sup.17 are independently
hydrogen or C.sub.1-3 alkyl-; or R.sup.15 and R.sup.16 taken
together with the carbon to which they are attached form a 3- to
7-membered cycloalkyl ring and R.sup.17 is hydrogen.
[0124] Preferably, the group [Linker]-R corresponds to (b) or (c)
above.
[0125] More preferably, the present invention provides a compound
of formula (I) or a pharmaceutically acceptable salt, N-oxide,
hydrate or solvate thereof, in which: [0126] ring A represents a
phenyl group, said ring A group being unsubstituted or substituted
by 1, 2 or 3 substituents which are the same or different and are
selected from halogen atoms and unsubstituted C.sub.1-4 alkyl,
C.sub.1-4 alkoxy, hydroxyl and --NR.sup.3R.sup.4 groups, wherein
R.sup.3 and R.sup.4 are the same or different and represent
hydrogen or unsubstituted C.sub.1-2 alkyl; [0127] R.sup.1 and
R.sup.2 each represent hydrogen; [0128] n is 1; [0129] X is NH;
[0130] V is --N.dbd.; [0131] W is --C(H).dbd.; and [0132] ring B
represents a 1,4-phenylene group, 1,3-phenylene group,
1,3-pyrazolyl group or pyridinyl group, in particular a
1,4-phenylene group, 1,3-phenylene group or 1,3-pyrazolyl group,
said ring B group being unsubstituted or substituted by 1, 2 or 3
substituents which are the same or different and are selected from
halogen atoms and unsubstituted C.sub.1-4 alkyl, C.sub.1-4 alkoxy,
hydroxyl and --NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4
are the same or different and represent hydrogen or unsubstituted
C.sub.1-2 alkyl.
[0133] In this embodiment of the invention, the group [Linker]-R
preferably corresponds to one of (a) to (c) as set out above,
preferably (b) or (c). More preferably, the group [Linker]-R
corresponds to (a'), (b') or (c') set out below:
(a') R represents a group of formula (X); [0134] R.sup.8 is as set
out below; [0135] one of R.sup.9 and R.sup.10 is hydrogen or
unsubstituted C.sub.1-4 alkyl and the other is an unsubstituted
C.sub.1-6 alkyl group; [0136] R.sup.11 represents a hydrogen atom
or an unsubstituted C.sub.1-2 alkyl group; [0137] [Linker]
represents --(CH.sub.2)--, wherein v is 1 or 2. (b') R represents a
group of formula (Y1'); [0138] R.sup.8 is as set out below; [0139]
[Linker] is --(CH.sub.2).sub.v-- or --C(.dbd.O)--, wherein v is 1
or 2. (c') R represents a group of formula (Y2); [0140] R.sup.8 is
as set out below; [0141] [Linker] is --C(.dbd.O)NR.sup.7--,
--CH.sub.2NR.sup.7-- or --CH.sub.2C(.dbd.O)NR.sup.7--, wherein
R.sup.7 represents hydrogen, unsubstituted methyl, unsubstituted
ethyl, unsubstituted isobutyl or --C(.dbd.O)CH.sub.3.
[0142] Typically, in this embodiment, when R is Y2, ring D is a 5-
to 7-membered saturated heterocyclyl group having at least one
nitrogen atom in the ring, wherein R.sup.8 and R.sup.11 are linked
to a ring carbon adjacent to a ring nitrogen, and wherein the bond
shown intersected by a wavy line is from a carbon atom in ring D,
with the proviso that D is not a pyrrolidinyl group. D may, for
example, be a pyrrolyl, piperidinyl, imidazolyl, oxazolyl,
isoxazolyl, thiazolyl, thiadiazolyl, pyrazolyl, pyridinyl,
pyridazinyl, pyrimidinyl, pyrazinyl, morpholinyl, piperazinyl
group, for instance a piperidinyl or piperazinyl group, for example
a piperidinyl group.
[0143] In this embodiment, preferably, ring B represents a
1,4-phenylene group, 1,3-phenylene group or pyridinyl group, in
particular a 1,4-phenylene or 1,3-phenylene group, said ring B
group being unsubstituted or substituted by 1, 2 or 3 substituents
which are the same or different and are selected from halogen atoms
and unsubstituted C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl and
--NR.sup.3R.sup.4 groups, wherein R.sup.3 and R.sup.4 are the same
or different and represent hydrogen or unsubstituted C.sub.1-2
alkyl. For example, ring B may represent a 1,4-phenylene or
1,3-phenylene group which is unsubstituted or substituted by 1, 2
or 3 substituents which are the same or different and are selected
from fluorine, chlorine, methyl and ethyl.
[0144] In each of the above (a') to (c'), R.sup.8 is a group --COOH
or an ester group of formula --(C.dbd.O)OR.sup.14 wherein R.sup.14
is R.sup.15R.sup.16R.sup.17C-- wherein either R.sup.15 is hydrogen
or C.sub.1-3alkyl-(Z.sup.1).sub.a--[(C.sub.1-C.sub.3)alkyl].sub.b-
wherein a and b are independently 0 or 1 and Z.sup.1 is --O--,
--S--, or --NH-- and R.sup.16 and R.sup.17 are independently
hydrogen or C.sub.1-3 alkyl-; or R.sup.15 and R.sup.16 taken
together with the carbon to which they are attached form a 3- to
7-membered cycloalkyl ring and R.sup.17 is hydrogen. Preferably
R.sup.8 is a group --COOH or an ester group of formula
--(C.dbd.O)OR.sup.14 wherein R.sup.14 is cyclopentyl or
t-butyl.
[0145] Preferably, the group [Linker]-R corresponds to (b') or (c')
above.
[0146] Particularly preferred compounds of the invention are:
[0147] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate [0148] Cyclopentyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piperazine-
-2-carboxylate [0149] Cyclopentyl
(2S)-4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piper-
azine-2-carboxylate [0150] Cyclopentyl
(2R)-4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piper-
azine-2-carboxylate [0151] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-chlorobe-
nzyl)amino]piperidine-2-carboxylate [0152] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(eth-
yl)amino]piperidine-2-carboxylate [0153] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(met-
hyl)amino]piperidine-2-carboxylate [0154] tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(eth-
yl)amino]piperidine-2-carboxylate [0155] Cyclopentyl
(2R)-4-[acetyl(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzy-
l)amino]piperidine-2-carboxylate [0156] Cyclopentyl
(2R)-4-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piper-
azine-2-carboxylate [0157] Cyclopentyl
1-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amino]cyc-
lobutanecarboxylate [0158] Cyclopentyl
N-[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]--
2-methylalaninate [0159] Cyclopentyl
(2S)-4-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piper-
azine-2-carboxylate [0160] Cyclopentyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-ethylbenzyl)pi-
perazine-2-carboxylate
Utilities
[0161] As mentioned above, the compounds with which the invention
is concerned are inhibitors of CSF-1R kinase, and are therefore of
use in the treatment of a variety of diseases. Accordingly, the
present invention provides a compound as defined herein for use in
a method of treatment of the human or animal body.
[0162] In particular, the present invention provides a compound as
defined herein for use in the treatment of a disease or disorder
mediated by CSF-1R kinase, including cell proliferative disease
such as cancer and psoriasis, polyglutamine disease such as
Huntingdon's disease, neurodegenerative disease such as Alzheimers
disease, autoimmune disease such as rheumatoid arthritis, diabetes,
haematological disease, inflammatory disease, cardiovascular
disease, atherosclerosis, and the inflammatory sequelia of
infection.
[0163] The present invention also provides a method of treating or
preventing a disease or disorder mediated by CSF-1R kinase in a
subject, which method comprises administering to said subject an
effective amount of a compound as defined herein, and use of a
compound as defined herein in the manufacture of a medicament for
use in the treatment or prevention of a disease or disorder
mediated by CSF-1R kinase. In particular, there is provided a
method of treating or preventing cell proliferative disease such as
cancer and psoriasis, polyglutamine disease such as Huntingdon's
disease, neurodegenerative disease such as Alzheimers disease,
autoimmune disease such as rheumatoid arthritis, diabetes,
haematological disease, inflammatory disease, cardiovascular
disease, atherosclerosis, and the inflammatory sequelia of
infection, by administering an effective amount of a compound of
formula (I) as defined herein. Also provided is use of a compound
of formula (I) as defined herein in the manufacture of a medicament
for use in the treatment or prevention of cell proliferative
disease such as cancer and psoriasis, polyglutamine disease such as
Huntingdon's disease, neurodegenerative disease such as Alzheimers
disease, autoimmune disease such as rheumatoid arthritis, diabetes,
haematological disease, inflammatory disease, cardiovascular
disease, atherosclerosis, and the inflammatory sequelia of
infection.
[0164] Autoimmune disease often has an inflammatory component. Such
conditions include acute disseminated alopecia universalise, ANCA
positive diseases, Behcet's disease, Chagas' disease, chronic
fatigue syndrome, dysautonomia, encephalomyelitis, ankylosing
spondylitis, aplastic anemia, hidradenitis suppurativa, autoimmune
hepatitis, autoimmune oophoritis, celiac disease, inflammatory
bowel disease, Crohn's disease, diabetes mellitus type 1, Fanconi
syndrome, giant cell arteritis, glomerulonephritis, Goodpasture's
syndrome, Grave's disease, Guillain-Barre syndrome, Hashimoto's
disease, Henoch-Schonlein purpura, Kawasaki's disease, systemic
lupus erythematosus, microscopic colitis, microscopic
polyarteritis, mixed connective tissue disease, multiple sclerosis,
myasthenia gravis, opsocionus myoclonus syndrome, optic neuritis,
Ord's thyroiditis, pemphigus, polyarteritis nodosa, polymyalgia,
rheumatoid arthritis, Reiter's syndrome, Sjogren's syndrome,
temporal arteritis, Wegener's granulomatosis, warm autoimmune
haemolytic anemia, interstitial cystitis, lyme disease, morphea,
psoriasis, sarcoidosis, scleroderma, ulcerative colitis, and
vitiligo.
[0165] Other inflammatory conditions which may be treated with the
compounds of the invention include, for example, appendicitis,
dermatitis, dermatomyositis, endocarditis, fibrositis, gingivitis,
glossitis, hepatitis, hidradenitis suppurativa, iritis, laryngitis,
mastitis, myocarditis, nephritis, otitis, pancreatitis, parotitis,
percarditis, peritonoitis, pharyngitis, pleuritis, pneumonitis,
prostatistis, pyelonephritis, and stomatisi, transplant rejection
(involving organs such as kidney, liver, heart, lung, pancreas
(e.g., islet cells), bone marrow, cornea, small bowel, skin
allografts, skin homografts, and heart valve xengrafts, sewrum
sickness, and graft vs host disease), acute pancreatitis, chronic
pancreatitis, acute respiratory distress syndrome, Sexary's
syndrome, congenital adrenal hyperplasis, nonsuppurative
thyroiditis, hypercalcemia associated with cancer, pemphigus,
bullous dermatitis herpetiformis, severe erythema multiforme,
exfoliative dermatitis, seborrheic dermatitis, seasonal or
perennial allergic rhinitis, bronchial asthma, contact dermatitis,
astopic dermatitis, drug hypersensistivity reactions, allergic
conjunctivitis, keratitis, herpes zoster ophthalmicus, iritis and
oiridocyclitis, chorioretinitis, optic neuritis, symptomatic
sarcoidosis, fulminating or disseminated pulmonary tuberculosis
chemotherapy, idiopathic thrombocytopenic purpura in adults,
secondary thrombocytopenia in adults, acquired (autoimmune)
haemolytic anemia, leukaemia and lymphomas in adults, acute
leukaemia of childhood, regional enteritis, autoimmune vasculitis,
multiple sclerosis, chronic obstructive pulmonary disease, solid
organ transplant rejection, sepsis, primary biliary cirrhosis and
primary sclerosing cholangitis.
[0166] Preferred treatments using compounds of the invention
include treatment of rheumatoid arthritis, psoriatic arthritis,
Type 1 diabetes, asthma, inflammatory bowel disease, systemic lupus
erythematosis, and inflammation accompanying infectious conditions
(e.g., sepsis), psoriasis, Crohns disease, ulcerative colitis,
chronic obstructive pulmonary disease, multiple sclerosis, atopic
dermatitis, and graft versus host disease.
[0167] Another preferred use of the compounds of the invention is
in the treatment of cancers.
[0168] The present invention also provides a pharmaceutical
composition which comprises a compound of formula (I) as defined
herein and one or more pharmaceutically acceptable carrier(s)
and/or excipients.
[0169] It will be understood that the specific dose level for any
particular subject will depend upon a variety of factors including
the activity of the specific compound employed, the age, body
weight, general health, sex, diet, time of administration, route of
administration, rate of excretion, drug combination and the
severity of the particular disease undergoing treatment. Optimum
dose levels and frequency of dosing will be determined by clinical
trial. However, it is expected that a typical dose will be in the
range from about 0.001 to 50 mg per kg of body weight.
[0170] The compounds with which the invention is concerned may be
prepared for administration by any route consistent with their
pharmacokinetic properties. The orally administrable compositions
may be in the form of tablets, capsules, powders, granules,
lozenges, liquid or gel preparations, such as oral, topical, or
sterile parenteral solutions or suspensions. Tablets and capsules
for oral administration may be in unit dose presentation form, and
may contain conventional excipients such as binding agents, for
example syrup, acacia, gelatin, sorbitol, tragacanth, or
polyvinyl-pyrrolidone; fillers for example lactose, sugar,
maize-starch, calcium phosphate, sorbitol or glycine; tabletting
lubricant, for example magnesium stearate, talc, polyethylene
glycol or silica; disintegrants for example potato starch, or
acceptable wetting agents such as sodium lauryl sulphate. The
tablets may be coated according to methods well known in normal
pharmaceutical practice. Oral liquid preparations may be in the
form of, for example, aqueous or oily suspensions, solutions,
emulsions, syrups or elixirs, or may be presented as a dry product
for reconstitution with water or other suitable vehicle before use.
Such liquid preparations may contain conventional additives such as
suspending agents, for example sorbitol, syrup, methyl cellulose,
glucose syrup, gelatin hydrogenated edible fats; emulsifying
agents, for example lecithin, sorbitan monooleate, or acacia;
non-aqueous vehicles (which may include edible oils), for example
almond oil, fractionated coconut oil, oily esters such as
glycerine, propylene glycol, or ethyl alcohol; preservatives, for
example methyl or propyl p-hydroxybenzoate or sorbic acid, and if
desired conventional flavouring or colouring agents.
[0171] For topical application to the skin, the drug may be made up
into a cream, lotion or ointment. Cream or ointment formulations
which may be used for the drug are conventional formulations well
known in the art, for example as described in standard textbooks of
pharmaceutics such as the British Pharmacopoeia.
[0172] For topical application by inhalation, the drug may be
formulated for aerosol delivery for example, by pressure-driven jet
atomizers or ultrasonic atomizers, or preferably by
propellant-driven metered aerosols or propellant-free
administration of micronized powders, for example, inhalation
capsules or other "dry powder" delivery systems. Excipients, such
as, for example, propellants (e.g. Frigen in the case of metered
aerosols), surface-active substances, emulsifiers, stabilizers,
preservatives, flavorings, and fillers (e.g. lactose in the case of
powder inhalers) may be present in such inhaled formulations. For
the purposes of inhalation, a large number of apparata are
available with which aerosols of optimum particle size can be
generated and administered, using an inhalation technique which is
appropriate for the subject. In addition to the use of adaptors
(spacers, expanders) and pear-shaped containers (e.g.
Nebulator.RTM., Volumatic.RTM.), and automatic devices emitting a
puffer spray (Autohaler.RTM.), for metered aerosols, in particular
in the case of powder inhalers, a number of technical solutions are
available (e.g. Diskhaler.RTM., Rotadisk.RTM., Turbohaler.RTM. or
the inhalers for example as described in European Patent
Application EP 0 505 321).
[0173] For topical application to the eye, the drug may be made up
into a solution or suspension in a suitable sterile aqueous or non
aqueous vehicle. Additives, for instance buffers such as sodium
metabisulphite or disodium edeate; preservatives including
bactericidal and fungicidal agents such as phenyl mercuric acetate
or nitrate, benzalkonium chloride or chlorhexidine, and thickening
agents such as hypromellose may also be included.
[0174] The active ingredient may also be administered parenterally
in a sterile medium. Depending on the vehicle and concentration
used, the drug can either be suspended or dissolved in the vehicle.
Advantageously, adjuvants such as a local anaesthetic, preservative
and buffering agent can be dissolved in the vehicle.
[0175] The compounds of the invention may be administered as part
of a pharmaceutical combination, which combination comprises
compounds of the invention and a second inhibitor. If administered
as part of a pharmaceutical combination, the compounds can either
be administered simultaneously or by any manner of separate
sequential administration of therapeutically effective amounts. It
does not matter if the compounds are administered in the same
dosage form, e.g., one compound may be administered topically and
the other compound may be administered orally. The administration
of a therapeutically effective amount of combinations of inhibitors
may be advantageous over the administration of a single inhibitor
in that the combination may provide one or more of the following
improved properties when compared to the individual administration
of a therapeutically effective amount of a component compound: i) a
greater therapeutic effect that the most active single agent, ii)
synergistic or highly synergistic activity, iii) a dosing protocol
that provides a reduced side effect profile, iv) a reduction in the
toxic effect profile, v) an increase in the therapeutic window, or
vi) an increase in the bioavailability of one or both of the
component compounds.
[0176] Examples of suitable pharmaceutical combinations include the
combination of compounds of the present invention with mTor
inhibitors, B-Raf inhibitors, AKT inhibitors or PI3 kinase
inhibitors.
Synthesis
[0177] There are multiple synthetic strategies for the synthesis of
the compounds (I) with which the present invention is concerned,
but all rely on known chemistry, known to the synthetic organic
chemist. Thus, compounds according to formula (I) can be
synthesised according to procedures described in the standard
literature and are well-known to those skilled in the art. Typical
literature sources are "Advanced organic chemistry", 4.sup.th
Edition (Wiley), J March, "Comprehensive Organic Transformation",
2.sup.nd Edition (Wiley), R. C. Larock, "Handbook of Heterocyclic
Chemistry", 2.sup.nd Edition (Pergamon), A. R. Katritzky), review
articles such as found in "Synthesis", "Acc. Chem. Res.", "Chem.
Rev", or primary literature sources identified by standard
literature searches online or from secondary sources such as
"Chemical Abstracts" or "Beilstein".
[0178] The compounds of the invention may be prepared by a number
of processes generally described below and more specifically in the
Examples hereinafter. In the reactions described below, it may be
necessary to protect reactive functional groups, for example
hydroxyl, amino and carboxy groups, where these are desired in the
final product, to avoid their unwanted participation in the
reactions [see for example Greene, T. W., "Protecting Groups in
Organic Synthesis", John Wiley and Sons, 1999]. Conventional
protecting groups may be used in conjunction with standard
practice. In some instances deprotection may be the final step in
the synthesis of a compound of general formula (I), and the
processes according to the invention described herein after are
understood to extend to such removal of protecting groups.
[0179] The schemes detailed below show general schemes for
synthesizing compounds of formula (I). It is recognised that the
compounds corresponding to Roman numerals in the schemes below do
not correspond to Roman numerals of claimed compounds employed in
other sections of this application.
[0180] Compounds of general formula (VII) may be prepared by, but
not limited to, the reactions set out in Scheme A.
##STR00008##
[0181] The compound with formula (II), 2-aminopyrazine, is
commercially available. 2-Aminopyrazine may be converted into the
compound of formula (III) by reaction with a brominating agent such
as N-bromosuccinimide in a suitable solvent such as DCM, at a
temperature of 0.degree. C.
[0182] Compounds of formula (IV) may be obtained from the compound
of formula (III) by selective nucleophilic aromatic substitution by
attack with benzylic amines. The reaction may be performed in a
suitable solvent such as n-butanol and in the presence of a base
such as DIPEA under microwave irradiation at a temperature of
150.degree. C.
[0183] Compounds of formula (V) can be obtained from compounds of
formula (IV) by reaction with appropriate aromatic boronic esters
or acids, using Suzuki coupling conditions. The Suzuki couplings
may be performed in the presence of a catalyst such as
dichlorobis(triphenylphosphine) palladium (II), a base such as
Na.sub.2CO.sub.3 and in a solvent such as DME at a temperature of
80.degree. C. It will be appreciated that to those skilled in the
art, various substituted heterocyclic and substituted aromatic
boronic esters and acids can be reacted with compounds of formula
(IV) under Suzuki coupling conditions to synthesise ring B
analogues of compounds of formula (V). Compounds of formula (VI)
can be obtained from compounds of formula (V) by reaction with a
base such as lithium hydroxide or sodium hydroxide in a solvent
such as ethanol, methanol, or THF and water at temperatures ranging
from RT to reflux. Compounds of formula (VI) can be treated with an
appropriate amine functionality and a suitable coupling agent, such
as 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide in a presence of
N-hydroxybenzotriazole, in an aprotic solvent such as
N,N-dimethylformamide at room temperature.
[0184] Compounds of general formula (IX) may be prepared by, but
not limited to, the reactions set out in Scheme B.
##STR00009##
[0185] Compounds of formula (IV) can be converted into compounds of
formula (VIII) using the conditions as described for the
preparation of compounds of formula (V) (Scheme A).
[0186] Compounds of formula (IX) may be prepared from compounds of
formula (VIII) under reductive amination conditions with a suitable
amine using an appropriate reducing agent such as sodium
triacetoxyborohydride in a suitable solvent such as DCE.
[0187] In the above Schemes A and B, the amino groups in the
compound of formula (VI) may first be protected by an appropriate
method for the protection of an amino group, known to those skilled
in the art. A suitable protecting group for an amino group is, but
not limited to, for example an acyl group, an alkanoyl group such
as acetyl, an alkoxycarbonyl group such as a methoxycarbonyl,
ethoxycarbonyl or tert-butoxycarbonyl group, an arylmethoxycarbonyl
group, such as benzyloxycarbonyl or an aroyl group, such as
benzoyl. The final stage of the reaction may then be an amino group
deprotection, using the necessary deprotection conditions which
vary depending with the choice of protecting group used. Thus, for
example, an acyl group such as alkanoyl, an alkoxycarbonyl group or
an aroyl group may be removed by hydrolysis with a suitable base
such as an alkali metal hydroxide, for example lithium hydroxide or
sodium hydroxide. Alternatively an acyl group such as a
tert-butoxycabonyl group may be removed, for example, by treatment
with a suitable acid such as hydrochloride, sulphuric or
trifluoroacetic acid and an arylmethoxycarbonyl group such as a
benzyloxycarbonyl group may be removed, for example, by
hydrogenation over a catalyst such as palladium on carbon.
[0188] Compounds of general formula (X) and (XI) may be prepared
by, but not limited to, the reactions set out in Scheme C.
##STR00010##
[0189] Compounds of formula (IX), when R.dbd.NHY2' can be converted
into compounds of formula (X) using the conditions as described for
the preparation of compounds of formula (IX) (Scheme B) with or
without the addition of an acid such as acetic acid. Alternatively
compounds of formula (IX), where R.dbd.NHY2', can be converted into
compounds of formula (XI) by an amide bond formation, by reaction
with acid chlorides in a suitable solvent such as DCM at
temperatures ranging of 0.degree. C.--room temperature.
[0190] Compounds of general formula (XVI) may be prepared by, but
not limited to, the reactions set out in Scheme D.
##STR00011##
[0191] Compounds of formula (IV) can be converted to compounds of
formula (XII) by an appropriate method for the protection of an
amino group, known to those skilled in the art. Suitable protecting
groups (Pg) are those described above.
[0192] Compounds as formula (XIII) may be obtained from compounds
of formula (XII) using the conditions as described for the
preparation of compounds of formula (V) (Scheme A). Compounds of
formula (XIII) may be transformed to compounds of formula (XIV) by
oxidation using an oxidising agent such as Dess-Martin periodinane
in an appropriate solvent such as DCM. Alternatively compounds of
formula (XIV) can be obtained directly from compounds of formula
(XII) by reaction with the appropriate substituted boronic acid or
ester using the conditions as described for the preparation of
compounds of formula (V) (Scheme A).
[0193] Compounds of formula (XV) may be prepared from compounds of
formula (XIV) by reductive amination using the conditions as
described for the preparation of compounds of formula (IX) (Scheme
B).
[0194] Compounds of formula (XVI) can be prepared by amino group
deprotection of the compounds of formula (XV) using the necessary
deprotection conditions which vary depending with the choice of
protecting group used, for example using one of the methods
described above.
[0195] A compound of formula (VII), (IX) or (XVI) in which R is Y1'
or NR.sup.7Y2'/NHY2' may be formed by (i) reacting a compound of
formula (VI), (VIII) or (XIV) according to Schemes A, B and D
respectively with an amino functionality in which the ring nitrogen
is substituted by a group R', for example tert-butoxycarbonyl, and
(ii) treating the resulting compound with a suitable acid such as
hydrochloric acid or trifluoroacetic acid.
[0196] A number of examples illustrative of the present invention
are described below. In most cases, alternative techniques could
also be used. In the following examples, it is understood that the
solvents and reagents used or suggested are not limiting and can be
substituted appropriately with solvents and reagents known to those
of skill in the art. Reaction products may be isolated by means
known in the art, such as extraction with a suitable solvent,
precipitation from a suitable solvent, chromatography using a
suitable solvent system, including silica gel column
chromatography, HPLC and the like.
[0197] The compounds of the invention may be synthesised by a
number of processes generally described above and more specifically
in the Examples hereinafter. Thus compounds of general formula (I)
may be prepared by, but not restricted to methods set out in
Schemes 1-30.
EXAMPLES
Abbreviations
[0198] Boc=tert-butoxycarbonyl Cs.sub.2CO.sub.3=cesium carbonate
DCE=1,2-dichloroethane DCM=dichloromethane
DIPEA=diisopropylethylamine DMAP=dimethylamino pyridine
DMF=dimethylformamide DMSO=dimethyl sulfoxide
EDC=1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
EtOAc=ethyl acetate HCl=hydrochloric acid
HOBt=1-hydroxybenzotriazole HPLC=high performance liquid
chromatography KOAc=potassium acetate LCMS=high performance liquid
chromatography/mass spectrometry MeCN=acetonitrile MeOH=methanol
MgSO.sub.4=magnesium sulphate Na.sub.2CO.sub.3=sodium carbonate
NaHCO.sub.3=sodium hydrogen carbonate NaOH=sodium hydroxide
NMR=nuclear magnetic resonance
PdCl.sub.2(dppf).sub.2=1,1'-Bis(diphenylphosphino)ferrocene-palladium(II)-
dichloride ppm=parts per million RT=room temperature STAB=sodium
triacetoxyborohydride TBME=tert-butyl methyl ether
THF=tetrahydrofuran hrs=hours mins=minutes g=gram(s)
mg=milligram(s) L=litre(s) mL=millilitre(s) .mu.L=mircolitre(s)
mol=mole(s) mmol=millimole(s) sat=saturated
[0199] Commercially available reagents and solvents (HPLC grade)
were used without further purification. Solvents were removed using
a Buchi rotary evaporator. Microwave irradiation was carried out
using a Biotage Initiator.TM. Eight microwave synthesiser.
Purification of compounds by flash chromatography column was
performed using silica gel, particle size 40-63 .mu.m (230-400
mesh) obtained from Fluorochem. Automated flash column
chromatography was performed on Combiflash.RTM. or Companion.RTM.
systems equipped with Telos, Biotage.RTM. or GraceResolv
cartridges. Purification of compounds by preparative HPLC was
performed on Gilson systems using reverse phase Waters XSelect C18
column (5 u, 50.times.19 mm) using the gradient method described
below.
[0200] Gradient 10-100% B (A=water/0.1% formic acid,
B=acetonitrile) over 10 min, flow=20 mL/min, UV detection at 254
nm. Alternatively, the formic acid can be replaced with
(NH.sub.4).sub.2CO.sub.3 in the aqueous buffer.
[0201] .sup.1H NMR spectra were recorded on a Bruker 300 MHz AV
spectrometer in deuterated solvents. Chemical shifts (8) are in
parts per million. Thin-layer chromatography (TLC) analysis was
performed with Kieselgel 60 F.sub.254 (Merck) plates and visualized
using UV light.
[0202] Analytical HPLC/MS was performed on an Agilent HP1100 LC
system using reverse phase Luna C18 columns (3 .mu.m, 50.times.4.6
mm), gradient 5-95% B (A=water/0.1% Formic acid,
B=acetonitrile/0.1% Formic acid) over 2.25 min, flow=2.25 mL/min.
UV spectra were recorded at 220 and 254 nm using a G1315B DAD
detector. Mass spectra were obtained over the range m/z 150 to 800
on a LC/MSD SL G1956B detector. Data were integrated and reported
using ChemStation and ChemStation Data Browser software.
INTERMEDIATES
Intermediate 1:
5-Bromo-N.sup.3-(2,6-dichlorobenzyl)pyrazine-2,3-diamine
##STR00012##
[0204] 5-Bromo-N.sup.3-(2,6-dichlorobenzyl)pyrazine-2,3-diamine was
synthesised using the route shown in Scheme 1.
##STR00013##
Stage 1. 3,5-Dibromopyrazin-2-amine
[0205] To a solution of aminopyrazine (11.36 g, 0.12 mol) in DCM
(700 mL) at 0.degree. C. was added N-bromosuccinimide (44.64 g,
0.25 mol) portion-wise. The reaction was stirred for 2 hrs. The
reaction was washed with sat Na.sub.2CO.sub.3 (3.times.200 mL),
dried over MgSO.sub.4, filtered and concentrated in vacuo before
purification by column chromatography (20% EtOAc/heptane) to give
the title compound as a yellow solid (15.9 g, 53%).
[0206] LCMS: m/z 252/254/256 [M+H].sup.+.
Stage 2.
5-Bromo-N.sup.3-(2,6-dichlorobenzyl)pyrazine-2,3-diamine
[0207] To a solution of 3,5-dibromopyrazin-2-amine (5.01 g, 19.8
mmol) in n-butanol (5 mL) was added 2, 6-dichlorobenzylamine (10.47
g, 59.4 mmol) and DIPEA (3.8 mL, 21.8 mmol). The reaction was
heated with microwaves at 150.degree. C. for 4 hrs. The reaction
was poured onto EtOAc (100 mL) and washed with water (50 mL), brine
(50 mL), dried over MgSO.sub.4, filtered and concentrated in vacuo.
The resulting solid was triturated with heptane and then DCM (40
mL) was added and the suspension was vigorously stirred for 15
mins. The solid was collected by filtration to give the title
compound as an off-white solid (5.14 g, 75%).
[0208] LCMS: m/z 347/349/381 [M+H].sup.+.
[0209] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 7.45-7.43
(2H, m), 7.32 (1H, dd, J=7.2, 8.9 Hz), 7.21 (1H, s), 4.81 (2H,
s).
Intermediate 2:
5-Bromo-N.sup.3-(2,6-difluorobenzyl)pyrazine-2,3-diamine
##STR00014##
[0211] 5-Bromo-N.sup.3-(2,6-diflorobenzyl)pyrazine-2,3-diamine was
synthesised in a similar manner to Intermediate 1 using 2,
6-difluorobenzylamine as the reactant in Stage 2 of the route shown
in Scheme 1. The product was purified by automated column
chromatography using EtOAc in heptane (gradient 0-40%).
[0212] LCMS: m/z 315/317 [M+H].sup.+.
[0213] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 7.49 (1H, s),
7.4-7.2 (1H, m), 7.07-6.85 (2H, m), 4.71 (2H, d, J=1.8 Hz),
4.70-4.55 (1H, m), 4.30 (2H, br, s).
Intermediate 3:
5-Bromo-N.sup.3-(2-chloro-6-fluorobenzyl)pyrazine-2,3-diamine
##STR00015##
[0215]
5-Bromo-N.sup.3-(2-chloro-6-fluorobenzyl)pyrazine-2,3-diamine was
synthesised in a similar manner to Intermediate 1 using
2-fluoro-6-chlorobenzylamine as the reactant in Stage 2 of the
route shown in Scheme 1. The product was purified by automated
column chromatography using EtOAc in heptane (gradient 0-70%).
[0216] LCMS: m/z 331/333 [M+H].sup.+.
[0217] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 7.6-7.47 (1H,
m), 7.42-7.17 (2H, m), 7.15-6.95 (1H, m), 4.78 (2H, dd, J=1.8, 0.5
Hz), 4.63-4.95 (1H, m), 4.06 (2H, br, s).
Intermediate 4: Di-tert-butyl
{5-bromo-3-[(tert-butoxycarbonyl)(2,6-dichloro
benzyl)amino]pyrazin-2-yl}imidodicarbonate
##STR00016##
[0219] Di-tert-butyl
{5-bromo-3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]pyrazin-2-yl}i-
midodicarbonate was prepared by the route shown in Scheme 2.
##STR00017##
Stage 1: Di-tert-butyl
{5-bromo-3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]pyrazin-2-yl}i-
midodicarbonate
[0220] To a solution of Intermediate 1 (3.76 g, 10.8 mmol) in DCE
(60 mL) at 60.degree. C. was added DMAP (0.13 g, 1.08 mmol)
followed by di-tert-butyl dicarbonate (14.1 g, 65.0 mmol) dissolved
in DCE (5 mL). The reaction mixture was stirred at 60.degree. C.
for 4 hrs for complete reaction. The reaction was allowed to cool
to RT before water (60 mL) was added. The organic layer was
separated and the aqueous layer was extracted with DCM (2.times.60
mL). The combined organics were dried over MgSO.sub.4, filtered and
concentrated in vacuo to an orange oil which was purified by column
chromatography (10% EtOAc/heptane) to give the title compound as a
yellow solid (3.98 g, 57%).
[0221] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 8.40 (1H, s),
7.32 (1H, m), 7.29 (1H, s), 7.18 (1H, dd, J=8.7, 6.9 Hz), 5.12 (2H,
s), 1.56 (18H, s), 1.47 (9H, s).
Intermediate 5:
3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde
##STR00018##
[0223] 3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde
was prepared by the route shown in Scheme 3 below.
##STR00019##
Stage 1.
3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde
[0224] To a solution of 3-formyl phenylboronic acid (5 g, 33 mmol)
in anhydrous THF (50 mL) was added pinacol (4.34 g, 37 mmol). The
reaction mixture was allowed to stir at RT under nitrogen for 18
hrs before concentration in vacuo. The crude residue was dissolved
in DCM (150 mL) and washed with water (3.times.100 mL). The organic
layer was dried over MgSO.sub.4 and evaporated in vacuo to give the
title compound as a yellow oil (7.73 g, 100%).
[0225] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 10.06 (1H,
s), 8.31 (1H, s), 8.07 (1H, d, J=7.4 Hz), 7.99 (1H, dt, J=1.5, 7.4
Hz), 7.54 (1H, t, J=7.4 Hz), 1.38 (12H, s).
Intermediate 6:
4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde
##STR00020##
[0227] 4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde
was synthesised in a similar manner to Intermediate 5 using
(4-formylphenyl)boronic acid as the starting material in Scheme
3.
[0228] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 10.06 (1H,
s), 7.97 (2H, d, J=8.1 Hz), 7.87 (2H, d, J=8.1 Hz), 1.37 (12H,
s).
Intermediate 7:
2-Ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde
##STR00021##
[0230]
2-Ethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde
was synthesised in a similar manner to Intermediate 5 using
3-ethyl-4-formylphenylboronic acid as the starting material in
Scheme 3.
[0231] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 10.34 (1H,
s), 7.85-7.74 (3H, m), 3.09 (2H, q, J=7.5 Hz), 1.38 (12H, s), 1.29
(3H, t, J=7.5 Hz).
Intermediate 8:
2-[3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]ethanol
##STR00022##
[0233]
2-[3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]ethanol
was prepared by the route shown in Scheme 4 below.
##STR00023##
Stage 1:
2-[3-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]ethanol
[0234] To a solution of 2-(3-bromophenyl)-ethanol (3.0 g, 15 mmol)
in anhydrous DMSO (30 mL) were added KOAc (4.39 g, 45 mmol),
bis(pinacolato)diboron (5.68 g, 22 mmol) and PdCl.sub.2(dppf).sub.2
(0.61 g, 0.74 mmol) and the reaction mixture was heated at
120.degree. C. under a nitrogen atmosphere for 18 hrs. The reaction
was cooled to RT and EtOAc (60 mL) was added. The reaction mixture
was filtered through Celite.RTM., washing with EtOAc (500 mL). The
filtrate was washed with sat NaHCO.sub.3 (150 mL), water (150 mL),
brine (150 mL), dried over MgSO.sub.4, filtered and concentrated in
vacuo. The crude material was purified by automated column
chromatography using EtOAc in heptane (gradient 0-100%) and then
column chromatography (33% EtOAc/heptane) to give the title
compound as a pale yellow oil (3.30 g, 89%).
[0235] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 7.70 (2H, m),
7.35 (2H, m), 3.89 (2H, t, J=6.6 Hz), 2.90 (2H, t, J=6.6 Hz), 1.51
(1H, s), 1.37 (12H, s).
Intermediate 9:
2-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzaldehyde
##STR00024##
[0237]
2-Methyl-4-(4,4,5,5-tetrameramethyl-1,3,2-dioxaborolan-2-yl)benzald-
ehyde was synthesised in a similar manner to Intermediate 8 using
4-bromo-2-methyl benzaldehyde as starting material in Scheme 4.
[0238] LCMS: m/z 430.2 [2M].sup.+.
[0239] .sup.1H NMR (300 MHz, CDCl.sub.3) b ppm: 10.32 (1H, s),
7.85-7.75 (2H, m), 7.72 (1H, s), 2.69 (3H, s), 1.38 (12H, s).
Intermediate 10: Methyl
[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]acetate
##STR00025##
[0241] Methyl
[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]acetate
was prepared according to the synthetic route shown in Scheme
5.
##STR00026##
[0242] To a solution of
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (560
mg, 2.9 mmol) in anhydrous DMF (10 mL) was added Cs.sub.2CO.sub.3
(1.03 g, 3.16 mmol) and methyl bromoacetate (0.30 mL, 3.16 mmol)
and the reaction was stirred at 90.degree. C. under a nitrogen
atmosphere for 18 hrs. The reaction was cooled, diluted with EtOAc
(50 mL), washed with water (2.times.25 mL), brine (25 mL), dried
over MgSO.sub.4, filtered and concentrated in vacuo. The crude
material was purified by column chromatography (50% EtOAc/heptane)
to give the title compound as a colourless oil (360 mg, 47%).
[0243] LCMS: m/z 267 [M+H].sup.+.
Intermediate 11:
2-[4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]ethano-
l
##STR00027##
[0245]
2-[4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl]-
ethanol was synthesised in a similar manner to Intermediate 10
using bromoethanol as the starting material in the route shown in
Scheme 5. The crude material was purified by column chromatography
(80-100% EtOAc/heptane) to give the title compound as a colourless
oil (528 mg, 49%).
[0246] LCMS: m/z 239 [M+H].sup.+.
Intermediate 12:
4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoic
acid
##STR00028##
[0248] -{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoic
acid was synthesised using the route shown in Scheme 6.
##STR00029##
Stage 1. Methyl
4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoate
[0249] To a solution of Intermediate 1 (3.82 g, 11.0 mmol) in DME
(75 mL) was added methyl
4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (4.48 g,
16.5 mmol) and 2N Na.sub.2CO.sub.3 (13.7 mL, 27.4 mmol) and the
solution was degassed by bubbling nitrogen through the reaction
mixture. Dichlorobis (triphenylphosphine) palladium (II) (385 mg,
0.5 mmol) was added and the reaction was stirred at 80.degree. C.
under a nitrogen atmosphere for 18 hrs. Another 0.05 eq of
dichlorobis (triphenylphosphine) palladium (II) (385 mg, 0.5 mmol)
was added and the reaction was stirred at 80.degree. C. under a
nitrogen atmosphere for 24 hrs for complete reaction. The reaction
mixture was filtered through Celite.RTM. and the filter cake was
washed with EtOAc (2.times.50 mL). The combined filtrates were
washed with water (50 mL), brine (50 mL), dried over MgSO.sub.4,
filtered and concentrated in vacuo before purification by column
chromatography (60% EtOAc/heptane) to give the title compound as a
yellow/green solid (3.88 g, 88%).
[0250] LCMS: m/z 403/405 [M+H].sup.+.
[0251] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 8.09-8.02
(4H, m), 7.82 (1H, s), 7.47-7.44 (2H, m), 7.36-7.29 (1H, m), 5.00
(2H, s), 3.92 (3H, s).
Stage 2.
4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoic
acid
[0252] To a suspension of methyl
4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoate (2.31
g, 5.7 mmol) in a 1:1:1 mixture of THF, MeOH and water (30 mL) was
added lithium hydroxide monohydrate (1.20 g, 28.6 mmol). The
reaction was stirred at RT for 20 hrs for complete reaction. The
solvent was removed under reduced pressure and the crude residue
was partitioned between water (100 mL) and EtOAc (50 mL). The
aqueous layer was separated and acidified to pH3 using 1N HCl.
[0253] The resulting solid was collected by filtration, washed with
water and dried in a vacuum oven at 40.degree. C. to give the title
compound as a yellow solid (1.43 g, 64%).
[0254] LCMS: m/z 389/391 [M+H].sup.+.
[0255] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 12.81 (1H,
br, s), 8.07-8.04 (2H, m), 7.95-7.93 (3H, m), 7.56-7.54 (2H, m),
7.41 (1H, dd, J=7.2, 8.9 Hz), 6.44 (1H, t, J=4.1 Hz), 6.40 (2H, br
s), 4.83 (2H, d, J=4.1 Hz).
Intermediate 13:
3-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoic
acid
##STR00030##
[0257] 3-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoic
acid was synthesised in a similar manner to Intermediate 12 using
the route shown in Scheme 6, reacting Intermediate 1 with
3-methoxycarbonylphenylboronic acid pinacol ester.
[0258] LCMS: m/z 389/391 [M+H].sup.+.
[0259] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 8.68-8.55
(1H, m), 8.12-8.05 (1H, m), 7.95-7.85 (1H, m), 7.78 (1H, m),
7.77-7.37 (3H, m), 7.35-7.29 (1H, m), 5.04 (2H, s).
Intermediate 14:
4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzaldehyde
##STR00031##
[0261]
4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzaldehyde
was synthesised in a similar manner to Intermediate 12 using Stage
1 of the route shown in Scheme 6, reacting Intermediate 1 with
Intermediate 6.
[0262] LCMS: m/z 373/375 [M+H].sup.+.
[0263] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 9.99 (1H,
s), 8.17 (2H, d, J=8.4 Hz), 9.01 (1H, s), 7.91 (2H, d, J=8.4 Hz),
7.57-7.54 (2H, m), 7.42 (1H, dd, J=7.1, 8.6 Hz), 6.49 (3H, br, s),
4.85 (2H, d, J=4.1 Hz).
Intermediate 15:
4-{5-Amino-6-[(2,6-difluorobenzyl)amino]pyrazin-2-yl}benzaldehyde
##STR00032##
[0265]
4-{5-Amino-6-[(2,6-difluorobenzyl)amino]pyrazin-2-yl}benzaldehyde
was synthesised in a similar manner to Intermediate 12 according to
Stage 1 of the route shown in Scheme 6, reacting Intermediate 2
with Intermediate 6.
[0266] LCMS: m/z 341 [M+H].sup.+.
Intermediate 16:
4-{5-Amino-6-[(2-chloro-6-fluorobenzyl)amino]pyrazin-2-yl}benzaldehyde
##STR00033##
[0268]
4-{5-Amino-6-[(2-chloro-6-fluorobenzyl)amino]pyrazin-2-yl}benzaldeh-
yde was synthesised in a similar manner to Intermediate 12
according to Stage 1 of the route shown in Scheme 6, reacting
Intermediate 3 with Intermediate 6.
[0269] LCMS: m/z 357/359 [M+H].sup.+.
[0270] .sup.1H NMR (300 MHz, CDCl.sub.3) ppm: 10.08 (1H, s),
8.3-7.85 (5H, m), 4.95 (2H, d J=1.6 Hz), 4.90-4.7 (1H, m),
4.69-4.40 (2H, m).
Intermediate 17:
4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-chlorobenzaldehyd-
e
##STR00034##
[0272]
4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-chlorobenza-
ldehyde was synthesised in a similar manner to Intermediate 12
according to Stage 1 of the route shown in Scheme 6, reacting
Intermediate 1 and 3-chloro-4-formyl phenylboronic acid.
[0273] LCMS: m/z 407/409 [M+H].sup.+.
[0274] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 10.51 (1H,
s), 8.12-7.94 (2H, m), 7.9-7.8 (1H, m), 7.48-7.0 (4H, m), 5.03 (2H,
d, J=1.6 Hz).
Intermediate 18:
4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-methylbenzaldehyd-
e
##STR00035##
[0276]
4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-methylbenza-
ldehyde was synthesised in a similar manner to Intermediate 12
according to Stage 1 of the route shown in Scheme 6, reacting
Intermediate 1 with Intermediate 9.
[0277] LCMS: m/z 387/389 [M+H].sup.+.
Intermediate 19:
4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-ethylbenzaldehyde
##STR00036##
[0279]
4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-ethylbenzal-
dehyde was synthesised in a similar manner to Intermediate 12
according to Stage 1 of the route shown in Scheme 6, reacting
Intermediate 1 with Intermediate 7.
[0280] LCMS: m/z 401/402 [M+H].sup.+.
Intermediate 20:
3-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzaldehyde
##STR00037##
[0282]
3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzaldehyde
was synthesised in a similar manner to Intermediate 12 using Stage
1 of the route shown in Scheme 6, reacting Intermediate 1 with
Intermediate 5.
[0283] LCMS: m/z 373/375 [M+H]+.sup.+.
[0284] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 10.13 (1H,
s), 8.6-8.5 (1H, m), 8.33-8.2 (1H, m), 8.03 (1H, m), 7.95-7.85 (1H,
m), 7.7-7.58 (1H, m), 7.49-73 (3H, m), 5.15-5.0 (2H, m), 4.6-4.4
(1H, m), 4.3-4.15 (2H, m).
Intermediate 21: Di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[3-(2-oxoethyl)phen-
yl]pyrazin-2-yl}imidodicarbonate
##STR00038##
[0286] Di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[3-(2-oxoethyl)phen-
yl]pyrazin-2-yl}imidodicarbonate was synthesised as shown in Scheme
7 below.
##STR00039##
Stage 1: Di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[3-(2-hydroxyethyl)-
phenyl]pyrazin-2-yl}imidodicarbonate
[0287] To a solution of Intermediate 4 (175 mg, 0.27 mmol) in DME
(2.6 mL), was added Intermediate 8 (100 mg, 0.40 mmol) and 2N
Na.sub.2CO.sub.3 (0.34 mL, 0.67 mmol). The solution was degassed by
bubbling nitrogen through the reaction mixture. Dichlorobis
(triphenylphosphine) palladium (II) (19 mg, 0.02 mmol) was added
and the reaction was stirred at 80.degree. C. under nitrogen for 18
hrs for complete reaction. The reaction mixture was filtered
through Celite.RTM. and the filter cake was washed with EtOAc (60
mL). The combined filtrates were washed with water (2.times.30 mL),
brine (30 mL), dried over MgSO.sub.4, filtered and concentrated in
vacuo before purification by automated column chromatography using
EtOAc in heptane (gradient 0-70%) to give the title compound as a
white solid (192 mg, 103%).
[0288] LCMS: m/z 695/697/699 [M+Na].sup.+.
Stage 2: Di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[3-(2-oxoethyl)phen-
yl]pyrazin-2-yl}imidodicarbonate
[0289] Di-tert-butyl {3-[(tert-butoxycarbonyl)
(2,6-dichlorobenzyl)amino]-5-[3-(2-hydroxyethyl)phenyl]pyrazin-2-yl}imido-
dicarbonate (169 mg, 0.25 mmol) was dissolved in anhydrous DCM (4
mL) and Dess-Martin periodinone (125 mg, 0.29 mmol) was added. The
reaction mixture was stirred at RT under nitrogen for 1.5 hrs. The
reaction was quenched with sat NaHCO.sub.3 (2 mL) and sat sodium
thiosulfate solution (2 mL) and vigorously stirred for 50 min. The
aqueous was separated and washed with DCM (3.times.10 mL). The
combined organics were dried over MgSO.sub.4, filtered and
concentrated in vacuo to give the title compound as a pale yellow
oil (174 mg, >100%). The material was not purified further.
[0290] LCMS: m/z 709/711/713 [M+H].sup.+.
Intermediate 22: 1-tert-Butyl 2-cyclopentyl
(2R)-4-aminopiperidine-1,2-dicarboxylate
##STR00040##
[0292] 1-tert-Butyl 2-cyclopentyl (2R)-4-am
inopiperidine-1,2-dicarboxylate was synthesised by the route shown
in Scheme 8.
##STR00041##
Stage 1. 1-tert-Butyl 2-cyclopentyl (2R)-4-oxopiperidine-1,
2-dicarboxylate
[0293] (2R)-1-(tert-Butoxycarbonyl)-4-oxopiperidine-2-carboxylic
acid (6 g, 24.7 mmol) was dissolved in DCM (100 mL) and cooled to
0.degree. C. Cyclopentanol (6.7 mL, 74 mmol), EDC (7.1 g, 37 mmol)
and DMAP (0.30 g, 2.5 mmol) added. The reaction was allowed to warm
to RT and stirred for 18 hrs. Saturated Na.sub.2CO.sub.3 was added
and the organic layer separated. The aqueous layer was extracted
with DCM (3.times.30 mL) and the combined organic layers were dried
over Na.sub.2SO.sub.4. The solvent was removed in vacuo and the
residue purified by automated column chromatography using EtOAc in
DCM (gradient 0-30%) to give the title compound (3.83 g, 97%).
[0294] LCMS: m/z 334 [M+H].sup.+.
Stage 2. 1-tert-Butyl 2-cyclopentyl
(2R)-4-(benzylamino)piperidine-1,2-dicarboxylate
[0295] 1-tert-Butyl 2-cyclopentyl
(2R)-4-oxopiperidine-1,2-dicarboxylate (3.82 g, 12.3 mmol) was
dissolved in DCM (40 mL) and benzylamine (1.4 mL, 12.9 mmol) was
added. To the reaction mixture was added acetic acid (0.84 mL, 14.7
mmol) and STAB (3.9 g, 18.4 mmol) and the reaction allowed to stir
at RT for 1 hr. The mixture was diluted with DCM (20 mL) and sat
NaHCO.sub.3 (100 mL) was added. The aqueous layer was extracted
with DCM (2.times.50 mL) and the combined organic layers were dried
over MgSO.sub.4, concentrated in vacuo and purified by automated
column chromatography using EtOAc in DCM (gradient 0-35%) to give
the title compound as a colourless oil (4.38 g, 84%).
[0296] LCMS: m/z 403.2 [M+H].sup.+.
Stage 3. 1-tert-Butyl 2-cyclopentyl
(2R)-4-aminopiperidine-1,2-dicarboxylate
[0297] 1-tert-Butyl 2-cyclopentyl
(2R)-4-(benzylamino)piperidine-1,2-dicarboxylate (4.38 g, 10 mmol)
was dissolved in methanol (40 mL) and charged with 10% Pd/C
catalyst (1.4 g) under a nitrogen atmosphere. The reaction was
stirred under a hydrogen balloon for 72 hrs for complete reaction.
The solution was filtered through Celite.RTM. and the filtrates
evaporated to dryness to give a colourless oil (3.7 g, 100%).
[0298] LCMS: m/z 313 [M+H].sup.+.
[0299] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 8.58 (2H, br,
s), 5.3-5.2 (1H, m), 4.65-4.52 (1H, m), 4.0-3.6 (2H, m), 3.42-3.15
(1H, m), 2.43-1.52 (12H, m), 1.46 (9H, s).
Intermediate 23: 1-tert-Butyl 2-cyclopentyl
(2S)-4-aminopiperidine-1,2-dicarboxylate
##STR00042##
[0301] 1-tert-Butyl 2-cyclopentyl
(2S)-4-aminopiperidine-1,2-dicarboxylate was synthesised in a
similar manner to Intermediate 22 using
(2S)-1-(tert-Butoxycarbonyl)-4-oxopiperidine-2-carboxylic acid as
the starting material in Scheme 8.
[0302] LCMS: m/z 313 [M+H].sup.+.
Intermediate 24: 1-tert-Butyl 2-cyclopentyl
(2R)-4-(methylamino)piperidine-1,2-dicarboxylate
##STR00043##
[0304] 1-tert-Butyl 2-cyclopentyl
(2R)-4-(methylamino)piperidine-1,2-dicarboxylate was synthesised in
a similar manner to Intermediate 22 using methylbenzylamine at
Stage 2 of the route shown in Scheme 8.
[0305] LCMS: m/z 327 [M+H].sup.+.
Intermediate 25: Di-tert-butyl
(2R)-4-aminopiperidine-1,2-dicarboxylate
##STR00044##
[0307] Di-tert-butyl (2R)-4-aminopiperidine-1,2-dicarboxylate was
synthesised in a similar manner to Intermediate 22 above starting
with di-tert-butyl (2R)-4-oxopiperidine-1,2-dicarboxylate at Stage
2 of Scheme 8. The synthesis of di-tert-butyl
(2R)-4-oxopiperidine-1,2-dicarboxylate is shown below in Scheme
9.
##STR00045##
Stage 1. Di-tert-butyl (2R)-4-oxopiperidine-1,2-dicarboxylate
[0308] (2R)-1-(tert-Butoxycarbonyl)-4-oxopiperidine-2-carboxylic
acid (10 g, 41.1 mmol) was dissolved in anhydrous toluene (100 mL)
and heated to 80.degree. C. under nitrogen. N,N-dimethylformamide
di-tert-butyl acetal (33.4 g, 164 mmol) was then added dropwise and
the reaction allowed to stir at 80.degree. C. for 2 hrs. The
reaction mixture was then cooled to RT and EtOAc (150 mL) and water
(150 mL) were added. The aqueous layer was extracted with EtOAc
(2.times.150 mL) and the combined organic layers purified by flash
column chromatography (20% EtOAc/heptane) to give the title
compound (8.82 g, 72%).
[0309] LCMS: m/z 322 [M+Na].sup.+.
Intermediate 26: Di-tert-butyl
(2S)-4-aminopiperidine-1,2-dicarboxylate
##STR00046##
[0311] Di-tert-butyl (2S)-4-aminopiperidine-1,2-dicarboxylate was
synthesised in a similar manner to Intermediate 22 above starting
with di-tert-butyl (2S)-4-oxopiperidine-1,2-dicarboxylate at Stage
2 of Scheme 8. Di-tert-butyl (2S)-4-oxopiperidine-1,2-dicarboxylate
was synthesised in a similar manner to di-tert-butyl
(2R)-4-oxopiperidine-1,2-dicarboxylate shown in Scheme 9.
[0312] LCMS: m/z 322 [M+Na].sup.+.
Intermediate 27: Di-tert-butyl
(2R)-4-(methylamino)piperidine-1,2-dicarboxylate
##STR00047##
[0314] Di-tert-butyl
(2R)-4-(methylamino)piperidine-1,2-dicarboxylate was synthesised in
a similar manner to Intermediate 22 starting at Stage 2 of the
route shown in Scheme 8 with di-tert-butyl
(2R)-4-oxopiperidine-1,2-dicarboxylate and reacting with
methylbenzylamine.
[0315] LCMS: m/z 315 [M+H].sup.+.
Intermediate 28: 1-tert-Butyl 2-cyclopentyl
piperazine-1,2-dicarboxylate
##STR00048##
[0317] 1-tert-Butyl 2-cyclopentyl piperazine-1,2-dicarboxylate was
synthesised using the route shown in Scheme 10.
##STR00049##
Stage 1: 4-Benzyl 1-tert-butyl 2-cyclopentyl
piperazine-1,2,4-tricarboxylate
[0318]
4-[(Benzyloxy)carbonyl]-1-(tert-butoxycarbonyl)piperazine-2-carboxy-
lic acid (9.96 g, 27 mmol) was dissolved in DCM (50 mL) and cooled
to 0.degree. C. Cyclopentanol (7.4 mL, 82 mmol), EDC (7.86 g, 41
mmol) and DMAP (0.33 g, 2.7 mmol) were then added and the reaction
allowed to warm to RT, and stirred for 24 hrs. Water (100 mL) and
DCM (50 mL) were then added and the layers separated. The aqueous
layer was re-extracted with DCM (2.times.50 mL) and the combined
organic layers were then dried over MgSO.sub.4 and concentrated in
vacuo. Purification by column chromatography (40% EtOAc/heptane)
afforded the title compound as a colourless oil (10.4 g, 88%).
[0319] LCMS: m/z 455 [M+Na].sup.+.
Stage 2. 1-tert-Butyl 2-cyclopentyl
piperazine-1,2-dicarboxylate
[0320] 4-Benzyl 1-tert-butyl 2-cyclopentyl
piperazine-1,2,4-tricarboxylate (10.4 g) was dissolved in EtOAc
(100 mL), and stirred with 10% palladium on carbon (2 g, 20% w/w)
under hydrogen at atmospheric pressure for 18 hrs. The reaction
mixture was then filtered through Celite.RTM. and the solvent
removed in vacuo to give the title compound as a white solid (6.62
g, 92%).
[0321] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 5.27 (1H, m),
4.50 (1H, m), 3.81 (1H, m), 3.50 (1H, m), 2.82-3.2 (3H, m), 2.72
(1H, m), 1.55-1.95 (8H, br m), 1.45 (9H, s).
Intermediate 29: 1-tert-Butyl 2-cyclopentyl
(2S)-piperazine-1,2-dicarboxylate
##STR00050##
[0323] 1-tert-Butyl 2-cyclopentyl (2S)-piperazine-1,2-dicarboxylate
was synthesised in a similar manner to Intermediate 28 using
(2S)-4-[(benzyloxy)carbonyl]-1-(tert-butoxycarbonyl)piperazine-2-carboxyl-
ic acid as the starting material in Scheme 10.
Stage 1. 4-Benzyl 1-tert-butyl 2-cyclopentyl
(2S)-piperazine-1,2,4-tricarboxylate
[0324]
(2S)-4-[(Benzyloxy)carbonyl]-1-(tert-butoxycarbonyl)piperazine-2-ca-
rboxylic acid (0.5 g, 1.37 mmol) was dissolved in dimethylformamide
(7 mL) and cooled to 0.degree. C. Cyclopentanol (0.25 mL, 2.74
mmol), EDC (0.395 g, 2.06 mmol) and DMAP (0.017 g, 0.14 mmol) were
added with stirring. The reaction was allowed to warm to room
temperature and was stirred for 23 hrs. The reaction was quenched
with water (40 mL) and the product extracted into EtOAc (3.times.20
mL). The combined organic layers were then washed with water
(2.times.20 mL) and brine (20 mL), before being dried over
MgSO.sub.4. The solvent was removed in vacuo and the residue
purified by automated column chromatography using EtOAc in heptane
(gradient 0-60%) to give the title compound (0.345 g, 58%).
[0325] LCMS: m/z 455 [M+Na].sup.+.
Stage 2. 1-tert-Butyl 2-cyclopentyl
(2S)-piperazine-1,2-dicarboxylate
[0326] 4-Benzyl 1-tert-butyl 2-cyclopentyl
(2S)-piperazine-1,2,4-tricarboxylate (0.35 g) was dissolved in
EtOAc (5 mL) and stirred with 10% palladium on carbon (0.069 g, 20%
w/w) under hydrogen at atmospheric pressure for 16.5 hrs. The
reaction mixture was then filtered through Celite.RTM. and the
solvent removed in vacuo to give the title compound (0.213 g,
90%).
[0327] LCMS: m/z 299 [M+H].sup.+.
[0328] .sup.1H NMR (300 MHz; CD.sub.3OD) .delta. ppm: 5.27 (1H, br,
s), 4.44-4.66 (1H, m), 3.71-3.89 (1H, m), 3.49 (1H, t, J=13.0 Hz),
2.84-3.16 (3H, m), 2.63-2.76 (1H, m), 1.87 (2H, br. s), 1.58-1.78
(6H, m), 1.47 (12H, d, J=8.7 Hz).
Intermediate 30: 1-tert-Butyl 2-cyclopentyl
(2R)-piperazine-1,2-dicarboxylate
##STR00051##
[0330] 1-tert-Butyl 2-cyclopentyl (2R)-piperazine-1,2-dicarboxylate
was synthesised in a similar manner to Intermediate 29 using
(2R)-4-[(benzyloxy)carbonyl]-1-(tert-butoxycarbonyl)piperazine-2-carboxyl-
ic acid as the starting material in Scheme 10.
[0331] LCMS: m/z 299 [M+H].sup.+.
[0332] .sup.1H NMR (300 MHz; CD.sub.3OD) .delta. ppm: 5.27 (1H, br,
s.), 4.41-4.67 (1H, m), 3.69-3.91 (1H, m), 3.49 (1H, t, J=12.6 Hz),
2.83-3.17 (3H, m), 2.63-2.77 (1H, m), 1.81-1.97 (2H, m), 1.58-1.78
(6H, m), 1.47 (12H, d, J=8.3 Hz).
Intermediate 31: Di-tert-butyl piperazine-1,2-dicarboxylate
##STR00052##
[0334] Di-tert-butyl piperazine-1,2-dicarboxylate was synthesised
using the route shown in Scheme 11.
##STR00053##
Stage 1. 4-Benzyl 1,2-di-tert-butyl
piperazine-1,2,4-tricarboxylate
[0335]
4-[(Benzyloxy)carbonyl]-1-(tert-butoxycarbonyl)piperazine-2-carboxy-
lic acid (1.0 g, 2.74 mmol) was dissolved in toluene (10 mL) and
heated to 80.degree. C. under a nitrogen atmosphere. DMF di
tert-butyl acetal was then added dropwise (2.6 mL, 11 mmol) and
stirring continued for 2 hrs. The reaction mixture was cooled and
partitioned between EtOAc (200 mL) and water (200 mL). The organic
layer was separated and the aqueous layer re-extracted with EtOAc
(2.times.100 mL). The combined organic layers were dried over
MgSO.sub.4, concentrated in vacuo and the residue was purified by
automated column chromatography (0-100% EtOAc/heptane) to give the
title compound as a colourless oil (1.08 g, 94%).
[0336] LCMS: m/z 443 [M+Na].sup.+.
Stage 2. Di-tert-butyl piperazine-1,2-dicarboxylate
[0337] 4-Benzyl 1,2-di-tert-butyl piperazine-1,2,4-tricarboxylate
(1.08 g) was dissolved in EtOAc (20 mL), and stirred with 10%
palladium on carbon (0.22 g, 20% w/w) under hydrogen at atmospheric
pressure for 17 hrs. The reaction mixture was then filtered through
Celite.RTM. and the solvent removed in vacuo to give the title
compound (0.810 g, >100%).
[0338] LCMS: m/z 287 [M+H].sup.+.
[0339] .sup.1H NMR (300 MHz; CD.sub.3OD) .delta. ppm: 3.72-3.96
(1H, m), 3.46-3.66 (1H, m), 2.98-3.22 (2H, m), 2.91 (1H, dd,
J=12.7, 4.4 Hz), 2.55-2.83 (3H, m), 1.40-1.55 (18H, m).
Intermediate 32: Cyclopentyl L-leucinate
##STR00054##
[0341] Cyclopentyl L-leucinate was prepared using the methodology
described below:
[0342] To a slurry of L-leucine (5 g, 30.5 mmol) in cyclohexane
(150 mL) were added cyclopentanol (27.5 mL, 305 mmol) and p-toluene
sulfonic acid (6.33 g, 33.3 mmol). The reaction was fitted with a
Dean-Stark receiver and heated to 135.degree. C. for complete
dissolution. This temperature was maintained for a period of 12 hrs
after which time the reaction was complete. The reaction was cooled
to RT with precipitation of a white solid. The solid was collected
by filtered and washed with EtOAc before drying under reduced
pressure. The required product was isolated as the tosylate salt
(10.88 g, 85%).
[0343] LCMS: m/z 200 [M+H].sup.+.
[0344] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 1.01 (6H, t,
J=5.8 Hz), 1.54-2.03 (11H, m), 2.39 (3H, s), 3.96 (1H, t, J=6.5
Hz), 5.26-5.36 (1H, m), 7.25 (2H, d, J=7.9 Hz), 7.72 (2H, d, J=8.3
Hz).
Intermediate 33: Cyclopenty 2-methylalaninate
##STR00055##
[0346] Cyclopentyl 2-methylaninate was prepared using the
methodology described below:
[0347] To a flask containing of 2-amino-2-methylpropionic acid (50
g, 0.48 mol) in cyclohexane (125 mL), cyclopentanol (267 mL, 2.91
mol) and p-toluene sulfonic acid (101.5 g, 0.53 mol) was fitted
with a Dean-Stark receiver and the reaction was heated to
135.degree. C. for 18 hrs. A further portion of p-toluene sulfonic
acid (34 g, 0.18 mol) was added and heated at reflux for 2 hrs
after which time the reaction was complete. The reaction was poured
onto ice-cold TBME (1 L) and the resulting solid was collected by
filtration. The solid was vigorously stirred in diethyl ether (2
L), filtered and dried in a vacuum oven to give the required
product as the tosylate salt (161 g, 97%).
[0348] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 8.36 (3H,
s), 7.48 (2H, d, J=8.1 Hz), 7.12 (2H, d, J=8.1 Hz), 5.18 (1H, t,
J=5.7 Hz), 2.29 (3H, s), 1.90-1.80 (2H, m), 1.75-1.55 (6H, m), 1.43
(6H, s).
Intermediate 34: Cyclopentyl 1-aminocyclobutanecarboxylate
##STR00056##
[0350] Cyclopentyl 1-aminocyclobutanecarboxylate was synthesised by
the route shown in Scheme 12.
##STR00057##
Stage 1. 1-[(tert-Butoxycarbonyl)amino]cyclobutanecarboxylic
acid
[0351] To a solution of 1-amino-1-cyclobutane carboxylic acid (2.92
g, 25.4 mmol), NaOH (3.04 g, 76.2 mmol), THF (100 mL) and water
(100 mL) at RT was added di-tert-butyl dicarbonate (8.30 g, 38.1
mmol) and the reaction was stirred for 24 hrs. The reaction mixture
was acidified to pH5 using 2M HCl and the mixture was extracted
with EtOAc (2.times.200 mL). The combined organics were dried over
MgSO.sub.4, filtered and concentrated in vacuo to give the title
compound as a white solid (1.05 g, 19%).
[0352] LCMS: m/z 238 [M+H].sup.+.
Stage 2. Cyclopentyl
1-[(tert-butoxycarbonyl)amino]cyclobutanecarboxylate
[0353] To a solution of
1-[(tert-butoxycarbonyl)amino]cyclobutanecarboxylic acid (5.2 g,
24.2 mmol), cyclopentanol (7.9 mL, 96.7 mmol) and DMAP (2.95 g,
24.2 mmol) in DCM (100 mL) was added EDC portion-wise (5.54 g, 29.0
mmol) and the reaction was stirred at RT for 3 hrs. The reaction
was diluted with DCM (200 mL) and the mixture washed with 0.5 M HCl
(2.times.150 mL), sat NaHCO.sub.3 (2.times.150 mL) and brine (100
mL).
[0354] The organic layer was separated, dried over MgSO.sub.4,
filtered, concentrated in vacuo and purified by automated column
chromatography using EtOAc in heptane (gradient 0-50%) to give the
title compound as a white solid (4.04 g, 59%).
[0355] LCMS: m/z 589 [M+Na].sup.+.
Stage 3. Cyclopentyl 1-aminocyclobutanecarboxylate
[0356] Cyclopentyl
1-[(tert-butoxycarbonyl)amino]cyclobutanecarboxylate (1.03 g, 3.63
mmol) was dissolved in 2N HCl in ether (25 mL) and stirred at RT
for 24 hrs. Further 2N HCl in ether (6 mL) was added and the
reaction was stirred for another 24 hrs. The solvent was
concentrated in vacuo to give the title compound as a white solid
(739 mg, 92%).
[0357] LCMS: m/z 184.2 [M+H].sup.+.
Intermediate 35: Cyclopentyl O-tert-butyl-L-serinate
##STR00058##
[0359] Cyclopentyl O-tert-butyl-L-serinate was synthesised by the
route shown in Scheme 13.
##STR00059##
Stage 1. Cyclopentyl
N-[(benzyloxy)carbonyl]-O-tert-butyl-L-serinate
[0360] N-[(Benzyloxy)carbonyl]-O-tert-butyl-L-serine (37.4 g, 0.13
mol), cyclopentanol (45.9 mL, 0.50 mol), EDC (29.1 g, 0.15 mol) and
DMAP (370 mg, 3.0 mmol) were stirred in DCM (370 mL) at RT for 18
hrs. Further cyclopentanol (23 mL, 0.25 mol) and DMAP (1.0 g, 8.2
mmol) were added and the reaction was stirred at RT for 6 hrs, then
30.degree. C. for 16 hrs. Water (500 mL) was added and the organic
layer was separated and concentrated in vacuo. The resulting
residue was purified by dry flash chromatography to give the title
compound (32.4 g, 69%).
[0361] .sup.1H NMR (300 Hz, CDCl.sub.3) .delta. ppm: 7.30-7.46 (5H,
m), 5.63 (1H, d, J=8.7 Hz), 5.19-5.30 (1H, m), 5.15 (2H, s), 4.42
(1H, dt, J=9.0, 3.2 Hz), 3.82 (1H, dd, J=8.7, 2.8 Hz), 3.57 (1H,
dd, J=8.7, 2.8 Hz), 1.51-1.96 (8H, m), 1.14 (9H, s).
Stage 2. Cyclopentyl O-tert-butyl-L-serinate
[0362] To solution of cyclopentyl
N-[(benzyloxy)carbonyl]-O-tert-butyl-L-serinate (32.4 g, 94.4 mmol)
in EtOAc (350 mL) was added 10% palladium on carbon catalyst (1.7
g, 5% w/w) under nitrogen. The reaction was stirred at RT for 16
hrs under a hydrogen atmosphere. The reaction was filtered through
Celite.RTM. and the filter cake was washed with EtOAc. The solvent
was concentrated in vacuo to give the title compound (22.3 g,
>100%).
[0363] .sup.1H NMR (300 Hz, CDCl.sub.3) .delta. ppm: 5.15 (1H, m),
3.37-3.60 (3H, m), 1.41-1.89 (10H, m), 1.09 (9H, s).
EXAMPLES
Example 1: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate
##STR00060##
[0365] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate was synthesised using the route shown
in Scheme 14.
##STR00061##
Stage 1. 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-1,2-dicarboxylate
[0366] Intermediate 12 (100 mg, 0.26 mmol) was dissolved in DMF (5
mL) and EDC (59 mg, 0.31 mmol) and HOBt (42 mg, 0.31 mmol) were
added. The reaction was stirred for 20 mins after which time
Intermediate 22 (96 mg, 0.31 mmol) was added. The reaction was
allowed to stir for 18 hours at RT. The reaction mixture was
diluted with EtOAc (50 mL) and washed with 2N NaOH (20 mL), water
(2.times.20 mL), brine (20 mL), dried over MgSO.sub.4, filtered and
concentrated in vacuo. The crude material was purified by column
chromatography (80% EtOAc/heptane) to give the title compound as a
yellow solid (95 mg, 54%).
[0367] LCMS: m/z 683/685 [M+H].sup.+.
Stage 2. Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate
[0368] 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-1,2-dicarboxylate (95 mg, 0.14 mmol) was dissolved in
DCM (20 mL) and 2M HCl in diethyl ether (2 mL) was added. The
resulting suspension was stirred at RT for 18 hours. The solvent
was removed in vacuo and the crude was dissolved in
Na.sub.2CO.sub.3 (20 mL) and the product was extracted with EtOAc
(3.times.20 mL). The combined organic layers were washed with brine
(20 mL), dried over MgSO.sub.4, filtered and concentrated in vacuo
in give a yellow oil which was dissolved in 1:4 MeCN/water (10 mL)
and freeze-dried to give the title compound as a yellow solid (63
mg, 78%).
[0369] LCMS: purity 95%, m/z 583/585 [M+H]r.
[0370] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 8.06 (2H, d,
J=8.5 Hz), 7.88 (2H, d, J=8.5 Hz), 7.81 (1H, s), 7.42 (2H, d, J=7.7
Hz), 7.35-7.29 (1H, m), 5.26-5.21 (1H, m), 4.15-4.04 (1H, m),
3.50-3.45 (1H, m), 3.24-3.18 (1H, m), 2.76 (1H, td, J=2.5, 12.6
Hz), 2.34 (1H, d, J=12.2 Hz), 1.98-1.41 (11H, m).
Example 2: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate
##STR00062##
[0372] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate was synthesised in a similar manner to
Example 1 using Intermediate 24 at Stage 1 in the route shown in
Scheme 14.
[0373] LCMS: purity 100%, m/z 597/599 [M+H].sup.+.
[0374] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 8.04 (2H, d,
J=8.1 Hz), 7.92 (1H, s), 7.46 (2H, d, J=8.1 Hz), 7.39-7.36 (2H, m),
7.25-7.19 (2H, m), 5.23 (1H, br s), 5.05 (2H, d, J=5.5 Hz), 4.51
(1H, t, J=5.4 Hz), 4.33 (2H, br s), 3.28 (1H, br s), 2.90 (4H, br
s), 1.90-1.60 (11H, m).
Example 3: Cyclopentyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)piperazin-
e-2-carboxylate
##STR00063##
[0376] Cyclopentyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)
piperazine-2-carboxylate was synthesised in a similar manner to
Example 1 using Intermediate 28 at Stage 1 in the route shown in
Scheme 14 with purification by preparative HPLC.
[0377] LCMS: purity 100%, m/z 569/571 [M+H].sup.+.
[0378] .sup.1H NMR (300 MHz, CDCl.sub.3) b ppm: 8.03 (2H, d, J=8.2
Hz), 7.92 (1H, s), 7.48 (2H, d, J=8.2 Hz), 7.38-7.35 (2H, m), 7.22
(1H, dd, J=7.4, 8.7 Hz), 5.24 (1H, br s), 5.04 (2H, d, J=5.5 Hz),
4.68 (1H, br, s), 4.56 (2H, br s), 3.53-3.31 (3H, m), 3.14-3.10
(1H, m), 2.84 (1H, br s), 2.18-1.58 (10H, m).
Example 4: tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate
##STR00064##
[0380] tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate was synthesised by the route shown in
Scheme 15.
##STR00065##
Stage 1. Di-tert-butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-1,2-dicarboxylate
[0381] Intermediate 12 (100 mg, 0.26 mmol) was dissolved in DMF (5
mL) and EDC (59 mg, 0.31 mmol) and HOBt (42 mg, 0.31 mmol) were
added. The reaction was stirred for 20 mins after which time
Intermediate 25 (96 mg, 0.31 mmol) was added. The reaction was
allowed to stir for 18 hours at RT. The reaction mixture was
diluted with EtOAc (50 mL) and washed with 2N NaOH (20 mL), water
(2.times.20 mL), brine (20 mL), dried over MgSO.sub.4, filtered and
concentrated in vacuo. The crude material was purified by column
chromatography (80% EtOAc/heptane) to give the title compound as a
yellow oil (153 mg, 89%).
[0382] LCMS: m/z 671/673 [M+H].sup.+.
Stage 2. tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate
[0383] Di-tert-butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-1,2-dicarboxylate (153 mg, 0.23 mmol) was dissolved
in dioxane (5 mL) and cooled to 0.degree. C. before the addition of
4M HCl in dioxane (20 mL). The reaction was stirred at 0.degree. C.
for 2 hrs. The reaction was basified to pH12 using 2N NaOH and
extracted with EtOAc (3.times.20 mL). The combined organic layers
were washed with brine (20 mL), dried over MgSO.sub.4, filtered and
concentrated in vacuo to give a yellow solid which was purified by
preparative HPLC to give the title compound as an off-white solid
(50 mg, 38%).
[0384] LCMS: purity 98%, m/z 571/573 [M+H].sup.+.
[0385] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 8.06 (2H, d,
J=8.5 Hz), 7.97 (1H, s), 7.82 (2H, d, J=8.5 Hz), 7.36-7.33 (2H, m),
7.19 (1H, dd, J=7.4, 8.6 Hz), 6.13 (1H, d, J=8.1 Hz), 5.05 (2H, d,
J=5.5 Hz), 4.61 (1H, t, J=5.5 Hz), 4.41 (2H, br s), 4.23-4.11 (1H,
m), 3.39 (1H, dd, J=2.7, 11.0 Hz), 3.26-3.19 (1H, m), 2.83-2.74
(1H, m), 2.44 (1H, d, J=12.4 Hz), 2.07-2.01 (1H, m), 1.47 (9H, s),
1.45-1.36 (2H, m).
Example 5: tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)(me-
thyl)amino]piperidine-2-carboxylate
##STR00066##
[0387] tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)(me-
thyl)amino]piperidine-2-carboxylate was synthesised in a similar
manner to Example 4 using Intermediate 27 at Stage 1 in the route
shown in Scheme 15.
[0388] LCMS: purity 96%, m/z 585/587 [M+H].sup.+.
[0389] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 8.04 (2H, d,
J=8.3 Hz), 7.96 (1H, s), 7.46 (2H, d, J=7.9 Hz), 7.38-7.36 (2H, m),
7.22 (1H, dd, J=7.4, 8.7 Hz), 5.05 (2H, d, J=5.5 Hz), 4.58 (1H, t,
J=5.5 Hz), 4.38 (2H, br s), 3.26 (2H, br s), 2.99-2.91 (4H, m),
2.19-1.98 (1H, m), 1.76 (2H, under H.sub.2O peak), 1.49 (9H,
s).
Example 6: tert-Butyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)piperazin-
e-2-carboxylate
##STR00067##
[0391] tert-Butyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)
piperazine-2-carboxylate was synthesised in a similar manner to
Example 4 using Intermediate 31 at Stage 1 in the route shown in
Scheme 15.
[0392] LCMS: purity 98%, m/z 557/559 [M+H].sup.+.
[0393] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 8.03 (2H, d,
J=8.2 Hz), 7.94 (1H, s), 7.48 (2H, d, J=8.2 Hz), 7.36-7.34 (2H, m),
7.20 (1H, dd, J=7.4, 8.9 Hz), 5.03 (2H, d, J=5.3 Hz), 4.61 (1H, t,
J=5.3 Hz), 4.43 (2H, br s), 3.44-3.37 (2H, m), 3.33-3.26 (1H, m),
3.12-3.08 (1H, m), 2.81 (1H, br s), 2.12 (2H, br s), 1.47 (9H,
s).
Example 7: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate
##STR00068##
[0395] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate was synthesised using the route shown in
Scheme 16.
##STR00069##
Stage 1: 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate
[0396] Intermediate 14 (150 mg, 0.40 mmol) and Intermediate 22 (138
mg, 0.44 mmol) were dissolved in DCE (15 mL) and stirred at RT for
2 hrs. STAB (170 mg, 0.80 mmol) was added and the reaction was
stirred for a further 18 hrs. The reaction was quenched by the
addition of sat Na.sub.2CO.sub.3 (20 mL) and vigorously stirred for
20 mins.
[0397] The aqueous layer was separated and further extracted with
DCM (2.times.20 mL) and the combined organics were dried over
MgSO.sub.4, filtered, concentrated in vacuo and purified by column
chromatography (80% EtOAc/heptane) to give the title compound as an
orange oil (168 mg, 62%).
[0398] LCMS: m/z 669/671 [M+H].sup.+.
Stage 2: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate
[0399] 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate (168 mg, 0.25 mmol) was dissolved in
DCM (20 mL) and 2M HCl in diethyl ether (5 mL) was added. The
mixture was stirred at RT for 18 hours. The solvent was removed in
vacuo and the crude was dissolved in Na.sub.2CO.sub.3 (20 mL) and
the product was extracted with EtOAc (3.times.20 mL). The combined
organic layers were washed with brine (20 mL), dried over
MgSO.sub.4, filtered and concentrated in vacuo to give a yellow oil
which was purified by preparative HPLC to afford the title compound
as an off-white solid (83 mg, 60%).
[0400] LCMS: purity 100%, m/z 569/571 [M+H].sup.+.
[0401] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 7.95 (2H, d,
J=8.1 Hz), 7.72 (1H, s), 7.48-7.41 (4H, m), 7.35-7.30 (1H, m),
5.23-5.18 (1H, m), 5.01 (2H, s), 3.91 (2H, br s), 3.18-3.14 (1H,
m), 2.81 (1H, br s), 2.67-2.59 (1H, m), 2.31 (1H, d, J=12.2 Hz),
2.00 (1H, d, J=12.8 Hz), 1.93-1.86 (2H, m), 1.85-1.60 (6H, m),
1.39-1.21 (2H, m), (1H missing. Under MeOH peak).
Example 8: Cyclopentyl
(2S)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate
##STR00070##
[0403] Cyclopentyl
(2S)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate was synthesised in a similar manner to
Example 7 using Intermediate 23 at Stage 1 in the route shown in
Scheme 16.
[0404] LCMS: purity 95%, m/z 569/571 [M+H].sup.+.
[0405] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 7.92 (2H, d,
J=8.1 Hz), 7.91 (1H, s), 7.41-7.33 (4H, m), 7.19 (1H, dd, J=7.3,
8.7 Hz), 5.24-5.18 (1H, m), 5.04 (2H, d, J=5.5 Hz), 4.51 (1H, t,
J=5.3 Hz), 4.35 (2H, br s), 3.90 (2H, s), 3.29 (1H, dd, J=2.8, 11.5
Hz), 3.24-3.18 (1H, m), 2.75-2.58 (2H, m), 2.29 (1H, d, J=13.2 Hz),
2.06-1.56 (9H, m), 1.33-1.20 (4H, m).
Example 9: Cyclopentyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piperazine-
-2-carboxylate
##STR00071##
[0407] Cyclopentyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
piperazine-2-carboxylate was synthesised in a similar manner to
Example 7 using Intermediate 28 at Stage 1 in the route shown in
Scheme 16.
[0408] LCMS: purity 99%, m/z 555/557 [M+H].sup.+.
[0409] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 7.96-7.91
(3H, m), 7.39-7.32 (4H, m), 7.17 (1H, dd, J=7.4, 8.6 Hz), 5.21 (1H,
t, J=5.8 Hz), 5.03 (2H, d, J=5.3 Hz), 4.57 (1H, t, J=5.3 Hz), 4.37
(2H, br s), 3.64-3.47 (3H, m), 3.11-3.05 (1H, m), 2.92-2.80 (2H,
m), 2.61-2.57 (1H, m), 2.46 (1H, t, J=8.9 Hz), 2.28 (1H, t, J=8.4
Hz), 2.04 (1H, br s), 1.90-1.81 (2H, m), 1.72-1.57 (6H, m).
Example 10: Cyclopentyl
(2S)-4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piper-
azine-2-carboxylate
##STR00072##
[0411] Cyclopentyl
(2S)-4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
piperazine-2-carboxylate was synthesised in a similar manner to
Example 7 using Intermediate 29 at Stage 1 in the route shown in
Scheme 16.
[0412] LCMS: purity 98%, m/z 555/557/559 [M+H].sup.+.
[0413] .sup.1H NMR (300 MHz, DMSO-de) .delta. ppm: 7.87 (2H, d,
J=7.9 Hz), 7.79 (1H, s), 7.56 (1H, s), 7.53 (1H, s), 7.37-7.44 (1H,
m), 7.28 (2H, d, J=8.1 Hz), 6.32 (1H, t, J=3.8 Hz), 6.18 (2H, s),
5.07 (1H, t, J=5.7 Hz), 4.82 (2H, d, J=3.8 Hz), 3.48-3.56 (1H, m),
3.36-3.42 (2H, m), 2.89-2.99 (1H, m), 2.53-2.69 (3H, m), 2.32-2.45
(2H, m), 2.19-2.30 (1H, m), 1.72-1.87 (2H, m), 1.44-1.63 (6H,
m).
Example 11: Cyclopentyl
(2R)-4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piper-
azine-2-carboxylate
##STR00073##
[0415] Cyclopentyl
(2R)-4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
piperazine-2-carboxylate was synthesised in a similar manner to
Example 7 using Intermediate 30 at Stage 1 in the route shown in
Scheme 16.
[0416] LCMS: purity 100%, m/z 555/557/559 [M+H].sup.+.
[0417] .sup.1H NMR (300 MHz, DMSO-de) .delta. ppm: 7.86 (2H, d,
J=8.1 Hz), 7.78 (1H, s), 7.49-7.56 (2H, m), 7.35-7.44 (1H, m), 7.28
(2H, d, J=8.1 Hz), 6.26 (1H, t, J=4.0 Hz), 6.08 (2H, s), 5.09 (1H,
t, J=5.8 Hz), 4.85 (2H, d, J=4.1 Hz), 3.36-3.58 (4H, m), 2.92-3.03
(1H, m), 2.60-2.75 (2H, m), 2.42 (2H, dd, J=10.3, 7.1 Hz),
2.22-2.33 (1H, m), 1.73-1.90 (2H, m), 1.43-1.68 (6H, m).
Example 12: Cyclopentyl
N-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te
##STR00074##
[0419] Cyclopentyl
N-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te was synthesised according to Stage 1 of the route shown in
Scheme 16 using the free base of Intermediate 32.
[0420] LCMS: purity 100%, m/z 556/558 [M+H].sup.+.
[0421] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 7.91 (2H, d,
J=8.3 Hz), 7.71 (1H, s), 7.47-7.44 (2H, m), 7.38-7.29 (3H, m),
5.22-5.16 (1H, m), 5.01 (2H, s), 3.81 (1H, d, J=13.0 Hz), 3.68 (1H,
d, J=13.0 Hz), 3.28 (1H, d, J=7.4 Hz), 1.94-1.82 (2H, m), 1.79-1.64
(7H, m), 1.53-1.48 (2H, m), 0.94 (3H, d, J=6.6 Hz), 0.89 (3H, d,
J=6.6 Hz).
Example 13: tert-Butyl
N-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te
##STR00075##
[0423] tert-Butyl
N-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te was synthesised according to Stage 1 of the route shown in
Scheme 16 using L-leucine tert-butyl ester as the reactant.
[0424] LCMS: purity 100%, m/z 544/546 [M+H].
[0425] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 7.90 (2H, d,
J=8.3 Hz), 7.69 (1H, s), 7.45-7.43 (2H, m), 7.38-7.27 (3H, m), 5.00
(2H, s), 3.80 (1H, d, J=12.9 Hz), 3.65 (1H, d, J=12.9 Hz), 3.20
(1H, t, J=7.3 Hz), 1.78-1.69 (1H, m), 1.50 (9H, s), 1.48-1.42 (2H,
m), 0.93 (3H, d, J=6.6 Hz), 0.88 (3H, d, J=6.6 Hz).
Example 14: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-difluorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate di-formate
##STR00076##
[0427] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-difluorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate was synthesised in a similar manner to
Example 7 starting with Intermediate 15 and reacting with
Intermediate 22 in Stage 1 in the route shown in Scheme 16. The
compound was isolated as the formate salt after preparative
HPLC.
[0428] LCMS: purity 98%, m/z 537 [M+H].sup.+.
[0429] .sup.1H NMR (300 MHz, DMSO-d.sub.5) .delta. ppm: 8.2 (2H,
s), 7.9-7.75 (2H, m), 7.7 (1H, s), 7.4-7.15 (3H, m), 7.12-6.95 (2H,
m), 6.65-6.5 (1H, m), 6.14 (2H, s), 5.1-5.0 (1H, m), 4.60 (2H, m),
3.7 (2H, m), 3.3-1.9 (5H, m), 1.70-1.65 (3H, m), 1.60-1.35 (6H,
m).
Example 15: Cyclopentyl
N-(4-{5-amino-6-[(2,6-difluorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te
##STR00077##
[0431] Cyclopentyl
N-(4-{5-amino-6-[(2,6-difluorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te was synthesised in a similar manner to Example 7 starting with
Intermediate 15 and reacting with the free base of Intermediate 32
according to Stage 1 of the route shown in Scheme 16.
[0432] LCMS: purity 98%, m/z 524 [M+H].sup.+.
[0433] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 7.84 (2H,
d, J=8.1 Hz), 7.77 (1H, s), 7.47-7.32 (1H, m), 7.28 (2H, d, J=8.3
Hz), 7.10 (2H, t, J=8.0 Hz), 6.63 (1H, t, J=5.0 Hz), 6.16 (2H, s),
5.11 (1H, t, J=5.8 Hz), 4.68 (2H, d, J=4.7 Hz), 3.82-3.4 (2H, m),
3.2-3.05 (1H, m), 2.40-2.25 (1H, m), 1.90-1.78 (2H, m), 1.77-1.50
(7H, m), 1.49-1.30 (2H, m), 0.83 (6H, d, J=2.2 Hz).
Example 16: tert-Butyl
N-(4-{5-amino-6-[(2,6-difluorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te
##STR00078##
[0435] tert-Butyl
N-(4-{5-amino-6-[(2,6-difluorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te was synthesised in a similar manner to Example 7 starting with
Intermediate 15 and reacting with L-leucine tert-butyl ester
according to Stage 1 of the route shown in Scheme 16.
[0436] LCMS: purity 98%, m/z 512 [M+H].sup.+.
[0437] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 7.85 (2H,
d, J=8.3 Hz), 7.77 (1H, s), 7.4-7.3 (1H, m), 7.29 (2H, d, J=8.1
Hz), 7.2-7.0 (2H, m), 6.7-6.6 (1H, m), 6.16 (2H, s), 4.68 (2H, d,
J=4.7 Hz), 3.9-3.5 (2H, m), 3.12-3.0 (1H, m), 2.3-2.1 (1H, m),
1.8-1.7 (1H, m), 1.44 (9H, s), 1.4-1.3 (2H, m), 0.84 (6H, d, J=2.2
Hz).
Example 17: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2-chloro-6-fluorobenzyl)amino]pyrazin-2-yl}benzyl-
)amino]piperidine-2-carboxylate
##STR00079##
[0439] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2-chloro-6-fluorobenzyl)amino]pyrazin-2-yl}benzyl-
)amino]piperidine-2-carboxylate was synthesised in a similar manner
to Example 7 starting with Intermediate 16 and reacting with
Intermediate 22 in Stage 1 in the route shown in Scheme 16.
[0440] LCMS: m/z 553 [M+H].sup.+.
[0441] .sup.1H NMR (300 MHz, DMSO-de) .delta. ppm: 7.83 (2H, d,
J=8.3 Hz), 7.76 (1H, s), 7.46-7.2 (5H, m), 6.55-6.4 (1H, m), 6.15
(2H, s), 5.12-5.0 (1H, m), 4.8-4.65 (2H, m), 3.72 (2H, br, s),
3.2-2.86 (2H, m), 2.6-2.3 (2H, m), 2.1-1.9 (3H, m), 1.85-1.70 (3H,
m), 1.65-1.45 (6H, m), 1.15-0.91 (2H, m).
Example 18: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-chlorobe-
nzyl)amino]piperidine-2-carboxylate
##STR00080##
[0443] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-chloro
benzyl)amino]piperidine-2-carboxylate was synthesised in a similar
manner to Example 7 starting with Intermediate 17 and reacting with
Intermediate 22 in Stage 1 in the route shown in Scheme 16.
[0444] LCMS: purity 98%, m/z 603/605 [M+H].sup.+.
[0445] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 7.95-7.88
(1H, m), 7.87-7.79 (2H, m), 7.60-7.48 (3H, m), 7.46-7.35 (1H, m),
6.48-6.38 (1H, s), 6.29 (2H, s), 5.13-5.0 (1H, m), 4.82 (2H, d,
J=4.3 Hz), 3.81 (2H, s), 3.30 (1H, m), 3.12 (1H, m), 2.95 (1H, m),
2.65-2.4 (2H, m), 2.08 (2H, m), 1.79 (3H, m), 1.70-1.45 (6H, m),
1.05 (2H, m).
Example 19: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-methylbe-
nzyl)amino]piperidine-2-carboxylate
##STR00081##
[0447] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-methylbe-
nzyl)amino]piperidine-2-carboxylate was synthesised in a similar
manner to Example 7 starting with Intermediate 18 and reacting with
Intermediate 22 in Stage 1 in the route shown in Scheme 16.
[0448] LCMS: purity 98%, m/z 583/585 [M+H].sup.+.
[0449] .sup.1H NMR (300 MHz, DMSO-de) .delta. ppm: 7.77 (1H, s),
7.73-7.63 (2H, m), 7.60-7.48 (2H, m), 7.45-7.2 (2H, m), 6.4-6.3
(1H, s), 6.14 (2H, s), 5.15-5.0 (1H, m), 4.82 (2H, d, J=4.0 Hz),
3.70 (2H, s), 3.45-2.9 (4H, m), 2.6-2.32 (3H, m), 2.34 (3H, s),
2.1-2.0 (1H, m), 1.90-1.75 (3H, m), 1.74-1.40 (5H, m) 1.15-0.95
(2H, m).
Example 20: Cyclopentyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-ethylbenzyl)pi-
perazine-2-carboxylate
##STR00082##
[0451] Cyclopentyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-ethylbenzyl)
piperazine-2-carboxylate was synthesised in a similar manner to
Example 7 starting with Intermediate 19 and reacting with
Intermediate 28 in Stage 1 in the route shown in Scheme 16.
[0452] LCMS: purity 98%, m/z 583/585 [M+H].sup.+.
[0453] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 7.77 (1H,
s), 7.76-7.74 (1H, m), 7.70-7.64 (1H, m), 7.60-7.50 (2H, m),
7.48-7.35 (1H, m), 7.24-7.18 (1H, m), 6.40-6.30 (1H, m), 6.17 (2H,
s), 5.10-5.00 (1H, m), 4.83 (2H, d, J=1.4 Hz), 3.50-3.40 (2H, m),
3.39-3.25 (2H, m under the water peak), 2.99-2.86 (1H, m), 2.71
(2H, q, J=2.5 Hz), 2.65-2.40 (3H, m under the DMSO peak), 2.38-2.22
(2H, m), 1.90-1.68 (2H, m), 1.67-1.40 (6H, m), 1.19 (3H, t, J=2.5
Hz).
Example 21: tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate
##STR00083##
[0455] tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate was synthesised as shown by the route in
Scheme 17.
##STR00084##
Stage 1. Di-tert-butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate
[0456] Intermediate 14 (100 mg, 0.27 mmol) and Intermediate 25 (89
mg, 0.29 mmol) were dissolved in DCE (10 mL) and stirred at RT for
2 hrs. STAB (114 mg, 0.29 mmol) was added and the reaction was
stirred for a further 18 hrs. The reaction was quenched by the
addition of sat Na.sub.2CO.sub.3 (20 mL) and vigorously stirred for
20 mins. The aqueous layer was separated and further extracted with
DCM (2.times.20 mL) and the combined organics were dried over
MgSO.sub.4, filtered, concentrated in vacuo and purified by column
chromatography (80% EtOAc/heptane) to give the title compound as an
orange oil (143 mg, 81%).
[0457] LCMS: m/z 657/659 [M+H].sup.+.
Stage 2. tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate
[0458] Di-tert-butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate (143 mg, 0.22 mmol) was dissolved in
dioxane (5 mL) and cooled to 0.degree. C. before the addition of 4M
HCl in dioxane (20 mL). The reaction was stirred at 0.degree. C.
for 2 hrs. The reaction was basified to pH12 using 2N NaOH and
extracted with EtOAc (3.times.25 mL). The combined organic layers
were washed with brine (20 mL), dried over MgSO.sub.4, filtered and
concentrated in vacuo to give a yellow oil, which was purified by
preparative HPLC to give the title compound as a pale yellow solid
(37 mg, 31%).
[0459] LCMS: purity 100%, m/z 557/559 [M+H].sup.+.
[0460] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 7.96-7.91
(3H, m), 7.41-7.33 (4H, m), 7.21-7.16 (1H, m), 5.04 (2H, d, J=5.5
Hz), 4.52 (1H, t, J=5.5 Hz), 4.31 (2H, br s), 3.90 (2H, s),
3.25-3.17 (2H, m), 2.72-2.57 (2H, m), 2.30 (1H, d, J=12.2 Hz), 1.94
(1H, d, J=11.9 Hz), 1.73 (2H, br, s), 1.47 (9H, s), 1.29-1.17 (2H,
m).
Example 22: tert-Butyl
(2S)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate
##STR00085##
[0462] tert-Butyl
(2S)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate was synthesised in a similar manner to
Example 21 using Intermediate 26 at Stage 1 in the route shown in
Scheme 17.
[0463] LCMS: purity 96%, m/z 557/559 [M+H].sup.+.
[0464] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 7.96 (2H, d,
J=8.3 Hz), 7.93 (1H, s), 7.45-7.35 (4H, m), 7.21 (1H, dd, J=7.3,
8.7 Hz), 5.04 (2H, d, J=5.5 Hz), 4.43 (1H, t, J=5.5 Hz), 4.22 (2H,
br, s), 3.91 (2H, s), 3.25-3.17 (2H, m), 2.75-2.58 (2H, m), 2.31
(1H, d, J=12.4 Hz), 1.96 (1H, d, J=12.4 Hz), 1.47 (9H, s),
1.35-1.21 (2H, s).
Example 23: tert-Butyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piperazine-
-2-carboxylate
##STR00086##
[0466] tert-Butyl
4-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piperazine-
-2-carboxylate was synthesised in a similar manner to Example 21
using Intermediate 31 at Stage 1 in the route shown in Scheme
17.
[0467] LCMS: purity 100%, m/z 543/545 [M+H].sup.+.
[0468] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 7.96-7.93
(3H, m), 7.41-7.37 (4H, m), 7.20 (1H, dd, J=7.3, 8.7 Hz), 5.04 (2H,
d, J=5.7 Hz), 4.45 (1H, t, J=5.5 Hz), 4.25 (2H, br, s), 3.64 (1H,
d, J=13.2 Hz), 3.51-3.46 (2H, m), 3.11-3.05 (1H, m), 2.93-2.82 (2H,
m), 2.61 (1H, d, J=11.3 Hz), 2.42 (1H, t, J=9.1 Hz), 2.26 (1H, t,
J=8.6 Hz), 1.94 (1H, br, s), 1.46 (9H, s).
Example 24: tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-difluorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate
##STR00087##
[0470] tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-difluorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate was synthesised in a similar manner to
Example 21 starting with Intermediate 15 and reacting with
Intermediate 25 in Stage 1 in the route shown in Scheme 17.
[0471] LCMS: purity 98%, m/z 525 [M+H].sup.+.
[0472] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 7.9-7.8
(2H, m), 7.76 (1H, s), 7.47-7.26 (3H, m), 7.18-7.0 (2H, m), 6.62
(1H, t, J=4.9 Hz), 6.14 (2H, s), 4.68 (2H, d, J=4.7 Hz), 3.73 (2H,
s), 3.2-2.85 (2H, m), 2.6-2.25 (2H, m), 2.15-2.00 (1H, m),
1.99-1.85 (1H, m), 1.80-1.60 (1H, m), 1.40 (9H, s), 1.14-0.90 (2H,
m).
Example 25: tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-chlorobe-
nzyl)amino]piperidine-2-carboxylate
##STR00088##
[0474] tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-chloro
benzyl)amino]piperidine-2-carboxylate was synthesised in a similar
manner to Example 21 starting with Intermediate 17 and reacting
with Intermediate 25 in Stage 1 in the route shown in Scheme
17.
[0475] LCMS: purity 96%, m/z 591/593 [M+H].sup.+.
[0476] .sup.1H NMR (300 MHz, DMSO-de) .delta. ppm: 7.98-7.88 (1H,
m), 7.87-7.75 (2H, m), 7.58-7.48 (3H, m), 7.45-7.35 (1H, m),
6.48-6.4 (1H, m), 6.29 (2H, s), 4.82 (2H, d, J=4.0 Hz), 3.82 (2H,
s), 3.32-3.26 (1H, m), 3.1-2.9 (2H, m), 2.5-2.4 (2H, m), 2.15-2.00
(1H, m), 1.99-1.85 (2H, m), 1.80-1.6 (1H, m), 1.39 (9H, s),
1.25-0.90 (2H, m).
Example 26: tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-methyl
benzyl)amino]piperidine-2-carboxylate
##STR00089##
[0478] tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-2-methylbe-
nzyl)amino]piperidine-2-carboxylate was synthesised in a similar
manner to Example 21 starting with Intermediate 18 and reacting
with Intermediate 25 in Stage 1 in the route shown in Scheme
17.
[0479] LCMS: purity 98%, m/z 571/573 [M+H].
[0480] .sup.1H NMR (300 MHz, DMSO-dc) .delta. ppm: 7.84-7.74 (1H,
m), 7.7-7.62 (2H, m), 7.59-7.45 (2H, m), 7.44-7.25 (2H, m),
6.41-6.24 (1H, m), 6.22-6.04 (2H, m), 4.91-4.7 (2H, m), 3.79-3.63
(2H, m), 3.6-3.2 (4H, m), 3.15-2.8 (2H, m), 2.5-2.4 (1H, m), 2.35
(3H, s), 2.2-2.0 (1H, m), 1.95-1.60 (1H, m), 1.39 (9H, s), 1.15-0.9
(1H, m).
Example 27: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(eth-
yl)amino]piperidine-2-carboxylate
##STR00090##
[0482] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
(ethyl)amino]piperidine-2-carboxylate was synthesised by the route
shown in Scheme 18.
##STR00091##
Stage 1: 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate
[0483] Intermediate 14 (150 mg, 0.40 mmol) and Intermediate 22 (138
mg, 0.44 mmol) were dissolved in DCE (15 mL) and stirred at RT for
2 hrs. STAB (170 mg, 0.80 mmol) was added and the reaction was
stirred for a further 18 hrs. The reaction was quenched by the
addition of sat Na.sub.2CO.sub.3 (20 mL) and vigorously stirred for
20 mins. The aqueous layer was separated and further extracted with
DCM (2.times.20 mL) and the combined organics were dried over
MgSO.sub.4, filtered, concentrated in vacuo and purified by column
chromatography (80% EtOAc/heptane) to give the title compound as an
orange oil (168 mg, 62%).
[0484] LCMS: m/z 669/671 [M+H].sup.+.
Stage 2: 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(eth-
yl)amino]piperidine-1,2-dicarboxylate
[0485] To a solution of 1-tert-butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate (168 mg, 0.28 mmol) in anhydrous
MeCN (4 mL) was added acetaldehyde (37 mg, 0.85 mmol). The reaction
was stirred at RT under nitrogen for 10 mins, followed by addition
of acetic acid (34 mg, 0.56 mmol) and STAB (132 mg, 0.62 mmol). The
reaction was stirred for 1 hour and then quenched by the addition
of sat NaHCO.sub.3 (10 mL). The aqueous was extracted with DCM
(3.times.20 mL) and the combined organics were dried over
MgSO.sub.4, filtered and concentrated in vacuo. The product was
obtained by column chromatography (50% EtOAc/heptane followed by
70% EtOAc/heptane) to give the title compound as a yellow solid (77
mg, 39%).
[0486] LCMS: m/z 697/698/699/700/701 [M+H].sup.+.
Stage 3: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
(ethyl)amino]piperidine-2-carboxylate
[0487] 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(eth-
yl)amino]piperidine-1,2-dicarboxylate (77 mg, 0.11 mmol) was
stirred in 2M HCl in diethyl ether (5 mL) at RT for 24 hours. The
solvent was removed in vacuo and the crude material was purified by
preparative HPLC to afford the title compound as an off-white solid
(32 mg, 49%).
[0488] LCMS: purity 100%, m/z 597/598/599/600 [M+H].sup.+
[0489] .sup.1H NMR (300 MHz, DMSO-de) .delta. ppm: 7.85 (2H, d,
J=8.1 Hz), 7.78 (1H, s), 7.49-7.59 (2H, m), 7.37-7.45 (1H, m), 7.33
(2H, d, J=8.1 Hz), 6.27-6.36 (1H, m), 6.15 (2H, s), 5.06 (1H, t,
J=5.7 Hz), 4.82 (2H, d, J=3.8 Hz), 3.59 (2H, s), 3.13 (1H, d,
J=10.9 Hz), 3.00 (1H, d, J=12.2 Hz), 2.66 (1H, t, J=11.7 Hz),
2.34-2.47 (2H, m), 2.03 (11H, br, s), 1.16-1.41 (2H, m), 0.95 (3H,
t, J=6.9 Hz).
Example 28: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(met-
hyl)amino]piperidine-2-carboxylate
##STR00092##
[0491] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(met-
hyl)amino]piperidine-2-carboxylate was synthesised by the route
shown in Scheme 19.
##STR00093##
Stage 1: 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate
[0492] Intermediate 14 (150 mg, 0.40 mmol) and Intermediate 22 (138
mg, 0.44 mmol) were dissolved in DCE (15 mL) and stirred at RT for
2 hrs. STAB (170 mg, 0.80 mmol) was added and the reaction was
stirred for a further 18 hrs. The reaction was quenched by the
addition of sat Na.sub.2CO.sub.3 (20 mL) and vigorously stirred for
20 mins. The aqueous layer was separated and further extracted with
DCM (2.times.20 mL) and the combined organics were dried over
MgSO.sub.4, filtered, concentrated in vacuo and purified by column
chromatography (80% EtOAc/heptane) to give the title compound as an
orange oil (168 mg, 62%).
[0493] LCMS: m/z 669/671 [M+H].sup.+.
Stage 2: 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(met-
hyl)amino]piperidine-1,2-dicarboxylate
[0494] To a solution of 1-tert-butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate (150 mg, 0.22 mmol) in MeCN (4 mL)
was added 37% aq. formaldehyde (16.8 .mu.L, 0.22 mmol). The
reaction was stirred at RT for 1.5 hrs, followed by addition of
STAB (71 mg, 0.34 mmol). The reaction was stirred for 3 hrs
followed by the further addition of 37% aq. formaldehyde (8.4
.mu.L, 0.11 mmol). The reaction was stirred for 18 hrs and then
quenched by the addition of sat NaHCO.sub.3 (20 mL). The aqueous
was extracted with EtOAc (4.times.20 mL) and the combined organics
were dried over MgSO.sub.4, filtered and concentrated in vacuo. The
crude was purified by automated column chromatography using EtOAc
in heptane (gradient 0-80%) to give the title compound, which
contained some impurities, as a yellow solid (125 mg, 83%).
[0495] LCMS: m/z 683/684/685 [M+H].sup.+.
Stage 3: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(met-
hyl)amino]piperidine-2-carboxylate
[0496] 1-tert-butyl 2-cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(met-
hyl)amino]piperidine-1,2-dicarboxylate (125 mg, 0.18 mmol) was
stirred in 2M HCl in diethyl ether (7 mL) at RT for 24 hours. The
solvent was removed in vacuo and the crude material was purified by
preparative HPLC to afford the title compound as an off-white solid
(13 mg, 12%).
[0497] LCMS: purity 100%, m/z 583/585/587 [M+H].sup.+.
[0498] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 7.86 (2H,
d, J=8.1 Hz), 7.79 (1H, s), 7.50-7.58 (2H, m), 7.36-7.46 (1H, m),
7.30 (2H, d, J=8.1 Hz), 6.28-6.35 (1H, m), 6.17 (2H, br, s),
5.03-5.12 (1H, m), 4.82 (2H, d, J=3.6 Hz), 3.56 (2H, s), 3.10-3.20
(1H, m), 2.96-3.08 (1H, m), 2.54-2.66 (1H, m), 2.33-2.45 (1H, m),
2.12 (3H, s), 1.90-2.07 (2H, m), 1.74-1.88 (2H, m), 1.46-1.71 (7H,
m), 1.19-1.44 (2H, m).
Example 29: Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(2-m-
ethylpropyl)amino]piperidine-2-carboxylate
##STR00094##
[0500] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(2-m-
ethylpropyl)amino]piperidine-2-carboxylate was synthesised by the
route shown in Scheme 20.
##STR00095##
Stage 1: Di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-(4-formylphenyl)pyr-
azin-2-yl}imidodicarbonate
[0501] To a solution of Intermediate 4 (569 mg, 0.88 mmol) in DME
(5.5 mL), was added Intermediate 6 (305 mg, 1.3 mmol) and 2N
Na.sub.2CO.sub.3 (1.1 mL, 2.2 mmol) and the solution was degassed
by bubbling nitrogen through the reaction mixture. Dichlorobis
(triphenylphosphine) palladium (II) (62 mg, 0.088 mmol) was added
and the reaction was stirred at 80.degree. C. under nitrogen for 18
hrs for complete reaction. The reaction mixture was filtered
through Celite.RTM. and the filter cake was washed with EtOAc (100
mL). The combined filtrates were washed with water (30 mL), brine
(30 mL), dried over MgSO.sub.4, filtered and concentrated in vacuo
before purification by column chromatography (20%-30%
EtOAc/heptane) to give the title compound as a colourless oil
(0.513 g, 87%).
[0502] LCMS: m/z 695/697/699 [M+Na].sup.+.
Stage 2: 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)(2,-
6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amino]piperidine-1,2-dicarboxyl-
ate
[0503] Di-tert-butyl {3-[(tert-butoxycarbonyl)
(2,6-dichlorobenzyl)amino]-5-(4-formylphenyl)
pyrazin-2-yl}imidodicarbonate (250 mg, 0.37 mmol) and Intermediate
22 (128 mg, 0.41 mmol) were dissolved in DCE (6 mL) and stirred at
RT, under nitrogen for 1 hr. STAB (157 mg, 0.74 mmol) was added and
the reaction was stirred for a further 18 hrs. The reaction was
quenched by the addition of sat Na.sub.2CO.sub.3 (10 mL) and DCM
(10 mL). The aqueous layer was separated and further extracted with
DCM (3.times.20 mL) and the combined organics were dried over
MgSO.sub.4, filtered, concentrated in vacuo and purified by column
chromatography (40% EtOAc/heptane) to give the title compound as an
orange oil (252 mg, 70%).
[0504] LCMS: m/z 969/970/971 [M+H].sup.+.
Stage 3: 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)(2,-
6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(2-methylpropyl)amino]piperidin-
e-1,2-dicarboxylate
[0505] 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)(2,-
6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amino]piperidine-1,2-dicarboxyl-
ate (80 mg, 0.08 mmol), isobutyraldehyde (113 .mu.L, 1.20 mmol) and
acetic acid (94 .mu.L, 1.6 mmol) were dissolved in DCE (3 mL) and
stirred at RT under nitrogen for 1 hr. STAB (35 mg, 0.33 mmol) was
added and the reaction was stirred 2.5 hrs. Further
isobutyraldehyde (75 .mu.L, 0.8 mmol) was added and the reaction
was stirred for 1.5 hrs, before further STAB (35 mg, 0.33 mmol) was
added. The reaction was stirred for 1.5 hrs and then quenched with
sat NaHCO.sub.3 (10 mL) and DCM (10 mL). The organic layer was
separated and the aqueous was extracted with DCM (3.times.10 mL).
The combined organics were dried over MgSO.sub.4, filtered,
concentrated in vacuo and purified by automated column
chromatography using EtOAc in heptane (gradient 0-30%) to give the
title compound as a colourless oil (70 mg, 83%).
[0506] LCMS: m/z 1025/1027/1029 [M+H].sup.+.
Stage 4: Cyclopentyl
(2R)-4-[acetyl(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzy-
l)amino]piperidine-2-carboxylate
[0507] 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)(2,-
6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
(2-methylpropyl)amino]piperidine-1,2-dicarboxylate (106 mg, 0.10
mmol) was stirred in dioxane (2 mL) and 4M HCl in dioxane (3 mL)
was added. The reaction was stirred for 18 hrs until completion.
The solvent was removed in vacuo to give a yellow solid, which was
purified by preparative HPLC to give the title compound as an
off-white solid (16 mg, 26%).
[0508] LCMS: purity 100%, m/z 625/627/629 [M+H].sup.+.
[0509] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm 7.86 (2H, d,
J=8.1 Hz), 7.79 (1H, s), 7.50-7.59 (2H, m), 7.37-7.45 (1H, m), 7.32
(2H, d, J=8.1 Hz), 6.27-6.36 (1H, m), 6.17 (2H, s), 5.06 (1H, t,
J=5.7 Hz), 4.82 (2H, d, J=3.8 Hz), 3.59 (2H, s), 3.08 (1H, d, J=9.8
Hz), 2.98 (1H, d, J=13.6 Hz), 2.54-2.64 (1H, m), 2.36 (1H, t,
J=10.9 Hz), 2.20 (2H, d, J=6.8 Hz), 2.05 (1H, br, s), 1.72-1.93
(3H, m), 1.48-1.71 (8H, m), 1.15-1.40 (2H, m), 0.81 (6H, d, J=6.4
Hz).
Example 30: tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(eth-
yl)amino]piperidine-2-carboxylate
##STR00096##
[0511] tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(eth-
yl)amino]piperidine-2-carboxylate was synthesised by the route
shown in Scheme 21.
##STR00097##
Stage 1. Di-tert-butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate
[0512] Intermediate 14 (100 mg, 0.27 mmol) and Intermediate 25 (89
mg, 0.29 mmol) were dissolved in DCE (10 mL) and stirred at RT for
2 hrs. STAB (114 mg, 0.29 mmol) was added and the reaction was
stirred for a further 18 hrs. The reaction was quenched by the
addition of sat Na.sub.2CO.sub.3 (20 mL) and vigorously stirred for
20 mins. The aqueous layer was separated and further extracted with
DCM (2.times.20 mL) and the combined organics were dried over
MgSO.sub.4, filtered, concentrated in vacuo and purified by column
chromatography (80% EtOAc/heptane) to give the title compound as an
orange oil (143 mg, 81%).
[0513] LCMS: m/z 657/659 [M+H].sup.+.
Stage 2: Di-tert-butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(eth-
yl)amino]piperidine-1,2-dicarboxylate
[0514] To a solution of di-tert-butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate (150 mg, 0.23 mmol) in MeCN (4 mL)
was added acetaldehyde (20 mg, 0.46 mmol). The reaction was stirred
at RT under nitrogen for 25 mins, followed by addition of acetic
acid (27 mg, 0.46 mmol) and STAB (106 mg, 0.50 mmol). The reaction
was stirred for 30 mins and then quenched by the addition of sat
NaHCO.sub.3 (10 mL). The aqueous was extracted with EtOAc
(3.times.20 mL) and the combined organics were dried over
MgSO.sub.4, filtered and concentrated in vacuo. The product was
obtained by automated column chromatography using EtOAc in heptane
(gradient 0-100%) to give the title compound as a yellow oil (83
mg, 53%).
[0515] LCMS: m/z 685/687/689 [M+H].sup.+.
Stage 3: tert-Butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)(eth-
yl)amino]piperidine-2-carboxylate
[0516] Di-tert-butyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
(ethyl)amino]piperidine-1,2-dicarboxylate (47 mg, 0.069 mmol) was
dissolved in DCM (0.5 mL) and cooled to 0.degree. C. before the
addition of 4M HCl in dioxane (2.5 mL). The reaction was stirred at
0.degree. C. for 1 hour. The reaction was basified to pH14 using 2N
NaOH (10 mL) and extracted with EtOAc (3.times.30 mL). The combined
organic layers were washed with brine (25 mL), dried over
MgSO.sub.4, filtered and concentrated in vacuo to give a yellow oil
which was purified by preparative HPLC to give the title compound
as a white solid (6 mg, 15%).
[0517] LCMS: purity 97.5%, m/z 585/587/589 [M+H].sup.+.
[0518] .sup.1H NMR (300 MHz, DMSO-d.sub.5) .delta. ppm: 7.85 (2H,
d, J=8.1 Hz), 7.77 (1H, s), 7.50-7.54 (2H, m), 7.36-7.43 (1H, m),
7.33 (2H, d, J=8.3 Hz), 6.24 (1H, t, J=4.2 Hz), 6.05 (2H, s), 4.85
(2H, d, J=4.3 Hz), 3.62 (2H, s), 2.98-3.09 (2H, m), 2.66 (1H, t,
J=11.7 Hz), 2.53 (2H, d, J=7.2 Hz), 2.37-2.47 (1H, m), 1.93 (1H, d,
J=12.1 Hz), 1.58-1.69 (1H, m), 1.41 (9H, s), 1.19-1.37 (2H, m),
0.96 (3H, t, J=7.0 Hz).
Example 31: Cyclopentyl
(2R)-4-[acetyl(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzy-
l)amino]piperidine-2-carboxylate
##STR00098##
[0520] Cyclopentyl
(2R)-4-[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
(ethyl)amino]piperidine-2-carboxylate was synthesised by the route
shown in Scheme 22.
##STR00099##
Stage 1: Di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-(4-formylphenyl)pyr-
azin-2-yl}imidodicarbonate
[0521] To a solution of Intermediate 4 (569 mg, 0.88 mmol) in DME
(5.5 mL), was added Intermediate 6 (305 mg, 1.3 mmol) and 2N
Na.sub.2CO.sub.3 (1.1 mL, 2.2 mmol) and the solution was degassed
by bubbling nitrogen through the reaction mixture. Dichlorobis
(triphenylphosphine) palladium (II) (62 mg, 0.088 mmol) was added
and the reaction was stirred at 80.degree. C. under nitrogen for 18
hrs for complete reaction. The reaction mixture was filtered
through Celite.RTM. and the filter cake was washed with EtOAc (100
mL). The combined filtrates were washed with water (30 mL), brine
(30 mL), dried over MgSO.sub.4, filtered and concentrated in vacuo
before purification by column chromatography (20%-30%
EtOAc/heptane) to give the title compound as a colourless oil
(0.513 g, 87%).
[0522] LCMS: m/z 695/697/699 [M+Na].sup.+.
Stage 2: 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)
(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amino]piperidine-1,2-dicarb-
oxylate
[0523] Di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-(4-formylphenyl)
pyrazin-2-yl}imidodicarbonate (250 mg, 0.37 mmol) and Intermediate
22 (128 mg, 0.41 mmol) were dissolved in DCE (6 mL) and stirred at
RT, under nitrogen for 1 hr.
[0524] STAB (157 mg, 0.74 mmol) was added and the reaction was
stirred for a further 18 hrs. The reaction was quenched by the
addition of sat Na.sub.2CO.sub.3 (10 mL) and DCM (10 mL). The
aqueous layer was separated and further extracted with DCM
(3.times.20 mL) and the combined organics were dried over
MgSO.sub.4, filtered, concentrated in vacuo and purified by column
chromatography (40% EtOAc/heptane) to give the title compound as an
orange oil (252 mg, 70%).
[0525] LCMS: m/z 969/970/971 [M+H].sup.+.
Stage 3: 1-tert-Butyl 2-cyclopentyl
(2R)-4-[acetyl(4-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbon-
yl)(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amino]piperidine-1,2-dica-
rboxylate
[0526] 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(4-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)
(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amino]piperidine-1,2-dicarb-
oxylate (134 mg, 0.14 mmol) was dissolved in anhydrous DCM (2 mL).
Triethylamine (21 mg, 0.21 mmol) was added and the reaction was
cooled to 0.degree. C. After 5 minutes acetyl chloride (16 mg, 0.27
mmol) was added and the reaction was warmed slowly to RT over 3
hrs. Additional acetyl chloride (8 mg, 0.10 mmol) was added and the
reaction was stirred for a further 2 hrs until completion. Sat
NaHCO.sub.3 (10 mL) and added and the product was extracted into
DCM (3.times.20 mL). The combined organics were dried over
MgSO.sub.4, filtered, concentrated in vacuo, and purified by
automated column chromatography using EtOAc in heptane (gradient
0-100%) to give the title compound as a colourless oil (107 mg,
77%).
[0527] LCMS: m/z 911/912/913/914 [M-Boc].sup.+.
Stage 4: Cyclopentyl
(2R)-4-[acetyl(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzy-
l)amino]piperidine-2-carboxylate
[0528] 1-tert-Butyl 2-cyclopentyl
(2R)-4-[acetyl(4-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbon-
yl)(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amino]piperidine-1,2-dica-
rboxylate (107 mg, 0.11 mmol) was dissolved in diethyl ether (0.5
mL) and 2M HCl in diethyl ether (10 mL) was added. The reaction was
stirred for 18 hrs until completion. The solvent was removed in
vacuo to give a yellow solid, which was purified by preparative
HPLC to give the title compound as an off-white solid (18 mg).
[0529] LCMS: purity 99%, m/z 611/613/615 [M+H].sup.+.
[0530] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 7.72-7.95
(3H, m), 7.48-7.56 (2H, m), 7.34-7.44 (1H, m), 7.16-7.27 (2H, m),
6.25 (1H, br, s), 6.01-6.13 (2H, m), 4.99-5.06 (1H, m), 4.85 (2H,
d, J=4.1 Hz), 4.56 (1.6H, s), 4.35-4.48 (1H, m), 3.91 (0.4H, br,
s), 3.24-3.33 (1H, m), 2.95 (1H, br, s), 2.19 (1H, br, s), 1.99
(3H, br, s), 1.68-1.81 (3H, m), 1.33-1.66 (10H, m).
Example 32: Cyclopentyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate
##STR00100##
[0532] Cyclopentyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate was synthesised by the route shown in
Scheme 23 below.
##STR00101##
Stage 1. 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-1,2-dicarboxylate
[0533] Intermediate 13 (100 mg, 0.26 mmol) was dissolved in DMF (6
mL) and Intermediate 22 (96 mg, 0.3 mmol) was added, followed by
HOBt (41 mg, 0.3 mmol) and EDC (56 mg, 0.3 mmol). The reaction
mixture was stirred for 18 hrs and then concentrated in vacuo. The
crude residue was taken forward unpurified (170 mg).
[0534] LCMS: m/z 683/685 [M+H].sup.+.
Stage 2. Cyclopentyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate
[0535] 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-1,2-dicarboxylate (170 mg, 0.24 mmol) was dissolved
in DCM (1 mL) and 2N HCl in diethyl ether (1 mL) added. The
reaction was stirred for 16 hrs and then concentrated in vacuo. The
crude residue was purified by preparative HPLC to give the title
compound as a yellow solid (50 mg).
[0536] LCMS: purity 97%, m/z 583/585 [M+H].sup.+.
[0537] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 8.43-8.3
(2H, m), 8.09 (1H, d, J=7.8 Hz), 7.91 (1H, s), 7.72 (1H, d, J=7.8
Hz), 7.65-733 (4H, m), 6.46-6.35 (1H, m), 6.28 (2H, s), 5.18-5.05
(1H, m), 4.84 (2H, d, J=4.0 Hz), 4.05-3.83 (1H, m), 3.5-2.4 (3H,
m), 2.2-2.0 (1H, m), 1.95-1.2 (12H, m).
Example 33: tert-Butyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate
##STR00102##
[0539] tert-Butyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzoyl)ami-
no]piperidine-2-carboxylate was synthesised in a similar manner to
Example 32 above using Intermediate 25 at Stage 1 of Scheme 23.
[0540] LCMS: purity 97%, m/z 571/573 [M+H].sup.+.
[0541] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 8.35 (2H,
m), 8.09 (1H, d, J=7.9 Hz), 7.91 (1H, s), 7.72 (1H, d, J=7.9 Hz),
7.60-7.34 (4H, m), 6.46-6.34 (1H, m), 6.28 (2H, s), 4.84 (2H, d,
J=4.1 Hz), 4.05-3.95 (1H, m) 3.3-2.4 (3H, m), 2.1-2.0 (2H, m),
1.7-1.6 (1H, m), 1.4 (9H, s) 1.45-1.25 (2H, m).
Example 34: Cyclopentyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate
##STR00103##
[0543] Cyclopentyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate was synthesised by the route shown in
Scheme 24 below.
##STR00104##
Stage 1. 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate
[0544] Intermediate 20 (78 mg, 0.2 mmol) was suspended in DCE (6
mL) and Intermediate 22 (97 mg, 0.31 mmol) added. The reaction
mixture was stirred for 10 mins before addition of STAB (85 mg, 0.4
mmol). The reaction mixture was allowed to stir at RT for 72 hrs
before addition of sat NaHCO.sub.3 (100 mL) and extraction of the
product with DCM (3.times.30 mL). The combined organic layers were
separated and dried over Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. Purification by automated column
chromatography (EtOAc/DCM) gave the product (115 mg) which still
contained minor impurity but was taken forward to Stage 2.
[0545] LCMS: m/z 669/671 [M+H].sup.+.
Stage 2. Cyclopentyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate
[0546] 1-tert-Butyl 2-cyclopentyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate (115 mg, 0.17 mmol) was dissolved in
DCM (1 mL) and 2N HCl in diethyl ether added. The reaction mixture
was stirred at RT for 18 hrs and then concentrated in vacuo. The
crude residue was purified by preparative HPLC to give the title
compound as an off-white solid (29 mg).
[0547] LCMS: purity 97%, m/z 569/571 [M+H].sup.+.
[0548] .sup.1H NMR (300 MHz, DMSO-de) .delta. ppm: 7.85-7.6 (3H,
m), 7.5-7.0 (5H, m), 6.32-6.23 (1H, m), 6.11 (2H, s), 5.1-4.9 (1H,
m), 4.8-4.7 (2H, m), 3.7 (2H, s), 3.15-2.9 (2H, m), 2.4-2.3 (1H,
m), 2.1-1.9 (3H, m), 1.85-1.65 (3H, m), 1.62-1.35 (7H, m), 1.1-0.9
(2H, m).
Example 35: Cyclopentyl
(2S)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate di-formate
##STR00105##
[0550] Cyclopentyl
(2S)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate was synthesised in a similar manner to
Example 34 using Intermediate 22 in Stage 1 of the synthetic route
shown in Scheme 24. Purification of the final compound was carried
out by preparative HPLC to afford the di-formate salt of the title
compound as a white solid (18.0 mg, 18%).
[0551] .sup.1H NMR (300 MHz, DMSO-d.sub.5) .delta. ppm 8.23 (2H,
s), 7.90 (1H, s), 7.85-7.78 (2H, m), 7.58-7.5 (2H, m), 7.48-7.2
(3H, m), 6.39-6.3 (1H, s), 6.20 (2H, s), 5.15-5.0 (1H, m), 4.83
(2H, d, J=4.1 Hz), 3.83 (2H, s), 3.5-2.8 (2H, m), 2.7-2.0 (3H, m),
1.9-1.4 (11H, m), 1.25-1.0 (2H, m)
[0552] LCMS: purity 98%, m/z 569/571 [M+H].sup.+.
Example 36: Cyclopentyl
(2S)-4-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piper-
azine-2-carboxylate
##STR00106##
[0554] Cyclopentyl
(2S)-4-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
piperazine-2-carboxylate was synthesised in a similar manner to
Example 34 using Intermediate 29 and DCM as solvent at Stage 1 in
the route shown in Scheme 24.
[0555] LCMS: purity 93%, m/z 555/557/559 [M+H].sup.+.
[0556] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 7.93 (1H, s),
7.83 (1H, d, J=7.9 Hz), 7.71 (1H, s), 7.40-7.46 (2H, m), 7.27-7.39
(2H, m), 7.20-7.27 (1H, m), 5.13-5.21 (1H, m), 5.03 (2H, s),
3.66-3.72 (1H, m), 3.53-3.66 (2H, m), 3.14-3.24 (1H, m), 2.86-2.97
(2H, m), 2.63-2.73 (1H, m), 2.57 (1H, dd, J=11.2, 8.0 Hz),
2.41-2.51 (1H, m), 1.71-1.90 (2H, m), 1.67 (1H, dd, J=6.8, 2.1 Hz),
1.44-1.64 (5H, m).
Example 37: Cyclopentyl
(2R)-4-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)piper-
azine-2-carboxylate
##STR00107##
[0558] Cyclopentyl
(2R)-4-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)
piperazine-2-carboxylate was synthesised in a similar manner to
Example 34 using Intermediate 30 and DCM as solvent at Stage 1 in
the route shown in Scheme 24.
[0559] LCMS: purity 100%, m/z 555/557/559 [M+H].sup.+.
[0560] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 7.92 (1H, s),
7.82 (1H, d, J=7.9 Hz), 7.71 (1H, s), 7.39-7.48 (2H, m), 7.20-7.38
(3H, m), 5.14 (1H, t, J=5.8 Hz), 5.02 (2H, s), 3.50-3.67 (3H, m),
3.05-3.15 (1H, m), 2.77-2.92 (2H, m), 2.58-2.68 (1H, m), 2.33-2.53
(2H, m), 1.71-1.89 (2H, m), 1.47-1.70 (6H, m).
Example 38: Cyclopentyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te
##STR00108##
[0562] Cyclopentyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te was prepared in a similar manner to Example 34 following Stage 1
of the route shown in Scheme 24 using Intermediate 32.
[0563] LCMS: purity 95%, m/z 556/558 [M+H].sup.+.
[0564] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 7.91-7.80
(2H, m), 7.81-7.78 (1H, m), 7.6-7.05 (5H, m), 5.22-5.10 (1H, m),
4.96 (2H, d, J=5.7 Hz), 4.38-4.22 (1H, m), 4.09 (2H, s), 3.81 (1H,
d, J=13.0 Hz), 3.61 (1H, d, J=13.0 Hz), 3.21 (1H, t, J=7.3 Hz),
1.9-1.3 (12H, m), 0.83 (3H, d, J=2.2 Hz), 0.78 (3H, d, J=2.2
Hz).
Example 39: tert-Butyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te
##STR00109##
[0566] tert-Butyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-L-leucina-
te was prepared in a similar manner to Example 34 following Stage 1
of the route shown in Scheme 24 and using tert-butyl
L-leucinate.
[0567] LCMS: purity 92%, m/z 544/546 [M+H].sup.+.
[0568] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 8.05-7.85
(3H, m), 7.45-7.15 (5H, m), 5.06 (2H, d, J=1.8 Hz), 4.5-4.4 (1H,
m), 4.19 (2H, s), 3.92 (1H, d, J=5.7 Hz), 3.72 (1H, d, J=5.7 Hz),
3.3-3.2 (1H, m), 1.9-1.8 (1H, m), 1.7-1.45 (3H, m), 1.58 (s, 9H),
0.94 (3H, d, J=2.2 Hz), 0.88 (3H, d, J=2.2 Hz).
Example 40: Cyclopentyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-2-methyla-
laninate
##STR00110##
[0570] Cyclopentyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-2-methyla-
laninate was prepared in a similar manner to Example 34 following
Stage 1 of the route shown in Scheme 24 and using Intermediate 33
(free base) and with final purification by preparative HPLC.
[0571] LCMS: purity 97%, m/z 528/530 [M+H].sup.+.
[0572] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 7.86 (1H,
s), 7.82-7.70 (2H, m), 7.6-7.5 (2H, m), 7.46-7.25 (3H, m), 6.4-6.3
(1H, m), 6.20 (2H, s), 5.3-5.0 (1H, m), 4.83 (2H, d, J=4.0 Hz),
3.60 (2H, s), 2.39-2.15 (1H, m), 1.94-1.4 (8H, m), 1.25 (6H,
s).
Example 41: tert-Butyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-2-methyla-
laninate formate salt
##STR00111##
[0574] tert-Butyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-2-methyla-
laninate was prepared in a similar manner to Example 34 following
Stage 1 of the route shown in Scheme 24 and using
alpha-aminoisobutyric acid tert-butyl ester and with final
purification by preparative HPLC to give the formate salt of the
title compound.
[0575] LCMS: purity 98%, m/z 516/518 [M+H].sup.+.
[0576] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 8.30 (1H,
br s), 7.95-7.86 (1H, m), 7.85-7.6 (2H, m), 7.62-7.47 (2H, m),
7.45-7.1 (3H, m), 6.40-6.30 (1H, m), 6.3-6.2 (2H, m), 4.9-4.8 (2H,
m), 3.7-3.58 (2H, m), 3.4-3.1 (1H, NH not visible underwater peak),
1.42 (9H, s), 1.23 (6H, s).
Example 42: Cyclopentyl
1-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amino]cyc-
lobutanecarboxylate di-formate
##STR00112##
[0578] Cyclopentyl
1-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amino]cyc-
lobutanecarboxylate was prepared in a similar manner to Example 34
following Stage 1 of the route shown in Scheme 18 and using
Intermediate 34 (free base). The final compound was purified by
preparative HPLC to afford the title compound as the di-formate
salt.
[0579] LCMS: purity 98%, m/z 540/541/542 [M+H].sup.+.
[0580] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 8.30 (2H,
br, s), 7.88 (1H, s), 7.82-7.75 (2H, m), 7.6-7.5 (2H, m), 7.46-7.17
(3H, m), 6.42-6.3 (1H, m), 6.21 (2H, s), 5.15-5.0 (1H, m), 4.83
(2H, d, J=4.1 Hz), 3.55 (2H, s), 2.35-2.15 (2H, m), 2.1-1.4 (13H,
m).
Example 43: Cyclopentyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-O-tert-bu-
tyl-L-serinate
##STR00113##
[0582] Cyclopentyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-O-tert-bu-
tyl-L-serinate was prepared in a similar manner to Example 34
following Stage 1 of the route shown in Scheme 24 and using
Intermediate 35 (free base). The final purification was done by
preparative HPLC.
[0583] LCMS: purity 98%, m/z 586/588 [M+H].sup.+.
[0584] .sup.1H NMR (300 MHz, DMSO-d) .delta. ppm: 7.88 (1H, s),
7.82-7.74 (2H, m), 7.6-7.5 (2H, m), 7.46-7.1 (3H, m), 6.43-6.3 (1H,
m), 6.21 (2H, b, s), 5.2-5.0 (1H, m), 4.83 (2H, d, J=1.3 Hz),
3.9-3.15 (5H, m), 2.45-2.25 (1H, m), 1.9-1.4 (8H, m), 1.04 (9H,
s).
Example 44: tert-Butyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-O-tert-bu-
tyl-L-serinate
##STR00114##
[0586] tert-Butyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-O-tert-bu-
tyl-L-serinate was prepared in a similar manner to Example 34
following Stage 1 of the route shown in Scheme 24 and using
tert-butyl O-tert-butyl-L-serinate hydrochloride. The final
purification was carried out by preparative HPLC.
[0587] LCMS: purity 98%, m/z 574/576 [M+H].sup.+.
[0588] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 7.89 (1H,
s), 7.84-7.72 (2H, m), 7.62-7.47 (2H, m), 7.45-7.12 (3H, m),
6.42-6.3 (1H, m), 6.21 (2H, s), 4.9-4.77 (2H, m), 3.83 (1H, d,
J=4.4 Hz), 3.64 (1H, d, J=4.4 Hz), 3.55-3.0 (3H, m), 2.42-2.2 (1H,
m), 1.39 (9H, s), 1.05 (9H, s)
Example 45: tert-Butyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-L-alanina-
te
##STR00115##
[0590] tert-Butyl
N-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)-L-alanina-
te was prepared in a similar manner to Example 34 according to
Stage 1 of the route shown in Scheme 24 and using tert-butyl
L-alaninate (free base) as the amino acid.
[0591] LCMS: purity 95%, m/z 502/504 [M+H].
[0592] .sup.1H NMR (300 MHz, DMSO-de) .delta. ppm: 7.89-7.82 (1H,
m), 7.81-7.7 (2H, m), 7.59-7.48 (2H, m), 7.46-7.1 (3H, m), 6.4-6.3
(1H, m), 6.20 (2H, br, s), 4.88-4.80 (2H, m), 3.86-3.5 (2H, m),
3.45-3.0 (1H, m), 2.45-2.2 (1H, m), 1.41 (9H, s), 1.16 (3H, d,
J=2.2 Hz).
Example 46: tert-Butyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate
##STR00116##
[0594] tert-Butyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate was synthesised as shown in Scheme
25.
##STR00117##
Stage 1. Di-tert-butyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate
[0595] Intermediate 20 (90 mg, 0.24 mmol) was suspended in DOE (4
mL) and Intermediate 25 (108 mg, 0.36 mmol) added. The reaction
mixture was stirred for 10 mins before addition of STAB (101 mg,
0.48 mmol). The reaction mixture was allowed to stir at RT for 18
hrs before addition of sat NaHCO.sub.3 (120 mL) and extraction of
the product with DCM (2.times.60 mL). The combined organic layers
were separated and dried over Na.sub.2SO.sub.4, filtered and
concentrated in vacuo. Purification by automated column
chromatography (EtOAc/DCM) gave the product (130 mg, 82%).
[0596] LCMS: m/z 657/659 [M+H].sup.+.
Stage 2. tert-Butyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate
[0597] Di-tert-butyl
(2R)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-1,2-dicarboxylate (130 mg, 0.197 mmol) was dissolved
in dioxane (3 mL) and cooled to 0.degree. C. before the dropwise
addition of 4M HCl in dioxane (1 mL). Over the course of 6 hours at
0.degree. C., an additional 2.5 mL of 4M HCL in dioxane was added
for complete reaction. To the reaction mixture was added 2N NaOH
(80 mL) and the product extracted with EtOAc (3.times.50 mL). The
combined organic layers were separated and dried over
Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The crude
residue was purified by preparative HPLC to give the title compound
as an orange solid (44 mg).
[0598] LCMS: purity 97%, m/z 557/559 [M+H].sup.+.
[0599] .sup.1H NMR (300 MHz, DMSO-de) .delta. ppm: 7.88 (1H, s),
7.85-7.8 (2H, m), 7.62-7.5 (2H, m), 7.47-7.15 (3H, m), 6.40-6.3
(1H, m), 6.18 (2H, s), 4.83 (2H, d, J=4.1 Hz), 3.78 (2H, s),
3.5-3.25 (2H, m), 3.11-2.88 (1H, m), 2.55-1.75 (5H, m), 1.38 (9H,
s), 1.15-0.9 (2H, m).
Example 47: tert-Butyl
(2S)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate di-formate
##STR00118##
[0601] tert-Butyl
(2S)-4-[(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}benzyl)amin-
o]piperidine-2-carboxylate was synthesised in a similar manner to
Example 46 using with Intermediate 26 in Stage 1 of the synthetic
route shown in Scheme 25. Purification of the final compound was
carried out by preparative HPLC to afford the di-formate salt of
the title compound as a white solid (19.8 mg, 8%).
[0602] LCMS: purity 96%, m/z 557/559 [M+H].sup.+
[0603] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. ppm: 8.27 (2H,
br, s), 7.9 (1H, s), 7.85-7.7 (2H, m), 7.7-7.47 (2H, m), 7.45-7.15
(3H, m), 6.4-6.3 (1H, m), 6.19 (2H, s), 4.88-4.75 (2H, m), 3.80
(2H, m), 3.75-3.2 (2H, m), 3.17-2.8 (2H, m), 2.65-2.3 (2H, m),
2.2-1.7 (2H, m), 1.38 (9H, s), 1.2-0.9 (2H, m).
Example 48: Cyclopentyl
N-[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]--
2-methylalaninate
##STR00119##
[0605] Cyclopentyl
N-[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]--
2-methylalaninate was synthesised as shown in Scheme 26 below.
##STR00120##
Stage 1: Cyclopentyl
N-[2-(3-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxy
carbonyl)
(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]-2-methylalaninate
[0606] Intermediate 21 (168 mg, 0.24 mmol) and Intermediate 33
(free base) (125 mg, 0.73 mmol) were dissolved in DCE (4 mL) and
stirred at RT for 10 mins under nitrogen. STAB (104 mg, 0.49 mmol)
was added and the reaction was stirred for a further 18 hrs. The
reaction was quenched by the addition of sat NaHCO.sub.3 (15 mL).
The aqueous layer was separated and further extracted with DCM
(3.times.20 mL) and the combined organics were dried over
MgSO.sub.4, filtered, concentrated in vacuo and purified by column
chromatography (50% EtOAc/heptane) to give the title compound as an
orange oil (64 mg, 31%).
[0607] LCMS: m/z 842/844/846 [M+H].sup.+.
Stage 2: Cyclopentyl
N-[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]--
2-methylalaninate
[0608] Cyclopentyl
N-[2-(3-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxy
carbonyl)(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]-2-methylala-
ninate (64 mg, 0.076 mmol) was stirred in 2M HCl in diethyl ether
(5 mL) was added. The reaction was stirred for 18 hrs. 4M HCl in
dioxane (2 mL) was added and the reaction was stirred for 30 mins
for complete reaction. The solvent was removed in vacuo to give a
yellow solid which was purified by preparative HPLC to give the
title compound as an off-white solid (15 mg, 36%).
[0609] LCMS: purity 100%, m/z 542/544/546 [M+H].sup.+.
[0610] .sup.1H NMR (300 MHz, DMSO-dc) .delta. ppm: 7.69-7.82 (3H,
m), 7.50-7.57 (2H, m), 7.36-7.45 (1H, m), 7.28 (1H, t, J=7.5 Hz),
7.07 (1H, d, J=7.2 Hz), 6.35 (1H, br. s), 6.19 (2H, s), 4.98 (1H,
t, J=5.5 Hz), 4.83 (2H, d, J=3.8 Hz), 2.60-2.77 (4H, m), 1.91 (1H,
br, s), 1.63-1.78 (2H, m), 1.36-1.61 (6H, m), 1.15 (6H, s).
Example 49: Cyclopentyl
1-{[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]-
amino}cyclobutanecarboxylate
##STR00121##
[0612] Cyclopentyl
1-{[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]-
amino}cyclobutanecarboxylate was synthesised in a similar manner to
Example 48 using 1.5 equivalents of Intermediate 34 in stage 1 of
the synthetic route shown in Scheme 26.
[0613] LCMS: purity 100%, m/z 554/556/558 [M+H].sup.+.
[0614] .sup.1H NMR (300 MHz, DMSO-d.sub.5) .delta. ppm: 7.69-7.82
(3H, m), 7.50-7.58 (2H, m), 7.35-7.44 (1H, m), 7.28 (1H, t, J=7.6
Hz), 7.08 (1H, d, J=7.5 Hz), 6.32-6.40 (1H, m), 6.19 (2H, s), 5.02
(1H, t, J=5.8 Hz), 4.83 (2H, d, J=4.1 Hz), 2.68-2.79 (2H, m), 2.59
(2H, br, s), 2.16-2.27 (2H, m), 2.12 (1H, br, s), 1.81-1.96 (3H,
m), 1.6-1.79 (3H, m), 1.38-1.60 (6H, m).
Example 50: Cyclopentyl
(2R)-4-{[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)e-
thyl]amino}piperidine-2-carboxylate
##STR00122##
[0616] Cyclopentyl
(2R)-4-{[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)e-
thyl]amino}piperidine-2-carboxylate was synthesised according to
the route shown in Scheme 27.
##STR00123##
Stage 1. 1-tert-Butyl 2-cyclopentyl
(2R)-4-{[2-(3-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)
(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]amino}piperidine-1,2--
dicarboxylate
[0617] Intermediate 21 (160 mg, 0.23 mmol), Intermediate 22 (109
mg, 0.35 mmol) and acetic acid (99 .mu.L, 1.7 mmol) were dissolved
in DCE (4 mL) and stirred at RT under nitrogen for 40 mins. STAB
(99 mg, 0.47 mmol) and acetic acid (0.2 mL, 3.5 mmol) were added
and the reaction was stirred for a further 18 hrs. The reaction was
quenched by the addition of sat NaHCO.sub.3 (10 mL) and DCM (20
mL). The aqueous layer was separated and further extracted with DCM
(3.times.10 mL) and the combined organics were dried over
MgSO.sub.4, filtered, concentrated in vacuo and purified by
automated column chromatography using EtOAc in heptane (gradient
0-50%) followed by column chromatography (40-50% EtOAc/heptane) to
give the title compound as a colourless oil (103 mg, 45%).
[0618] LCMS: m/z 983/985/987 [M+H].sup.+.
Stage 2. Cyclopentyl
(2R)-4-{[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)e-
thyl]amino}piperidine-2-carboxylate
[0619] To a solution of 1-tert-butyl 2-cyclopentyl
(2R)-4-{[2-(3-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)-
(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]amino}piperidine-1,2-d-
icarboxylate (100 mg, 0.10 mmol) in dioxane (2 mL) was added 4M HCl
in dioxane (3 mL) and the reaction was stirred at RT for 21 hrs.
The solvent was concentrated in vacuo and the resulting residue was
purified by preparative HPLC to give the title compound as a pale
orange oil (10 mg, 17%).
[0620] LCMS: purity 95.6%, m/z 583/585/587 [M+H].sup.+.
[0621] 1H NMR (300 MHz, DMSO): .delta. ppm: 7.70-7.81 (3H, m),
7.50-7.58 (2H, m), 7.35-7.44 (1H, m), 7.28 (1H, t, J=7.5 Hz), 7.10
(1H, d, J=7.5 Hz), 6.36 (1H, t, J=4.2 Hz), 6.18 (2H, s), 5.01-5.10
(1H, m), 4.83 (2H, d, J=4.1 Hz), 3.14 (1H, dd, J=11.4, 2.5 Hz),
2.94 (1H, dt, J=12.6, 3.2 Hz), 2.67-2.85 (4H, m), 2.54-2.59 (1H,
m), 2.36-2.47 (1H, m), 1.47-2.07 (12H, m), 0.87-1.09 (2H, m).
Example 51: Cyclopentyl
(2S)-4-{[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)e-
thyl]amino}piperidine-2-carboxylate
##STR00124##
[0623] Cyclopentyl
(2S)-4-{[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)e-
thyl]amino}piperidine-2-carboxylate was synthesised in a similar
manner to Example 50 using Intermediate 23 in stage 1 of the route
shown in Scheme 27.
[0624] LCMS: purity 94%, m/z 583/585/587 [M+H].sup.+.
[0625] .sup.1H NMR (300 MHz, DMSO-de) .delta. ppm: 7.79 (1H, s),
7.70-7.77 (2H, m), 7.51-7.57 (2H, m), 7.37-7.43 (1H, m), 7.28 (1H,
t, J=7.6 Hz), 7.09 (1H, d, J=7.5 Hz), 6.36 (1H, t, J=4.2 Hz), 6.19
(2H, s), 5.05 (1H, t, J=6.1 Hz), 4.82 (2H, d, J=4.1 Hz), 3.14 (1H,
dd, J=11.3, 2.1 Hz), 2.89-2.97 (1H, m), 2.67-2.84 (4H, m), 2.54
(1H, br, s), 2.36-2.47 (1H, m), 2.01 (2H, d, J=10.9 Hz), 1.68-1.85
(3H, m), 1.44-1.67 (7H, m), 0.89-1.07 (2H, m).
Example 52: Cyclopentyl
4-[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]p-
iperazine-2-carboxylate
##STR00125##
[0627] Cyclopentyl
4-[2-(3-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}phenyl)ethyl]p-
iperazine-2-carboxylate was synthesised in a similar manner to
Example 50 using Intermediate 28 in stage 1 of the route shown in
Scheme 27.
[0628] LCMS: purity 97%, m/z 569/571/573 [M+H].sup.+.
[0629] .sup.1H NMR (300 MHz, DMSO-de) .delta. ppm: 7.70-7.81 (3H,
m), 7.49-7.57 (2H, m), 7.35-7.44 (1H, m), 7.27 (1H, t, J=7.6 Hz),
7.10 (1H, d, J=7.5 Hz), 6.33-6.40 (1H, m), 6.20 (2H, s), 5.02-5.11
(1H, m), 4.83 (2H, d, J=4.1 Hz), 3.39 (2H, d, J=3.0 Hz), 2.86-2.98
(1H, m), 2.67-2.81 (3H, m), 2.52-2.67 (3H, m), 2.32-2.47 (2H, m),
2.16-2.31 (1H, m), 1.70-1.85 (2H, m), 1.43-1.68 (6H, m).
Example 53: Cyclopentyl
(2R)-4-{[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetyl]amino}piperidine-2-carboxylate
##STR00126##
[0631] Cyclopentyl
(2R)-4-{[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetyl]amino}piperidine-2-carboxylate was synthesised as
shown in Scheme 28.
##STR00127##
Stage 1:
(4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetic acid
[0632] To a solution of Intermediate 1 (1.34 g, 3.8 mmol) in DME
(20 mL), was added Intermediate 10 (956 mg, 4.2 mmol) and 2N
Na.sub.2CO.sub.3 (4.8 mL, 9.6 mmol) and the solution was degassed
by bubbling nitrogen through the reaction mixture. Dichlorobis
(triphenylphosphine) palladium (II) (270 mg, 0.4 mmol) was added
and the reaction was stirred at 90.degree. C. under nitrogen for 18
hrs. The reaction mixture was concentrated in vacuo and the residue
was dissolved in water (100 mL) and washed with EtOAc (50 mL). The
aqueous was filtered and acidified to pH3 using 1N HCl. The
resulting solid was collected by filtration, washed with water and
dried in a vacuum oven to give the title compound as a beige solid
(400 mg, 26%).
[0633] LCMS: m/z 393/395 [M+H].
Stage 2: 1-tert-Butyl 2-cyclopentyl
(2R)-4-{[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetyl]amino}piperidine-1,2-dicarboxylate
[0634]
(4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazol-1-
-yl)acetic acid (100 mg, 0.26 mmol) was dissolved in DMF (5 mL) and
EDC (59 mg, 0.31 mmol) and HOBt (41 mg, 0.31 mmol) were added. The
reaction was stirred for 20 mins after which time Intermediate 22
(95 mg, 0.31 mmol) was added. The reaction was allowed to stir for
72 hours at RT. The reaction mixture was diluted with EtOAc (25 mL)
and washed with 2N NaOH (10 mL), water (10 mL), brine (10 mL),
dried over MgSO.sub.4, filtered and concentrated in vacuo. The
crude material was purified by column chromatography (5%
MeOH/EtOAc) to give the title compound as a yellow oil (122 mg,
70%).
[0635] LCMS: m/z 687/689 [M+H].sup.+.
Stage 3: Cyclopentyl
(2R)-4-{[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetyl]amino}piperidine-2-carboxylate
[0636] 1-tert-Butyl 2-cyclopentyl
(2R)-4-{[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetyl]amino}piperidine-1,2-dicarboxylate (122 mg, 0.18
mmol) was dissolved in DCM (5 mL) and 2N HCl in diethyl ether (20
mL) was added. The reaction mixture was stirred at RT for 1 hrs and
then concentrated in vacuo. The crude residue was partitioned
between sat Na.sub.2CO.sub.3 (20 mL) and EtOAc (20 mL). The
organics were separated and concentrated in vacuo before
purification by preparative HPLC to give the title compound as an
off-white solid (41 mg, 39%).
[0637] LCMS: purity 98%, m/z 587/589 [M+H].sup.+.
[0638] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 8.03 (1H, s),
7.92 (1H, s), 7.68 (1H, s), 7.37-7.34 (2H, m), 7.23-7.18 (1H, m),
6.36 (1H, d, J=7.9 Hz), 5.21-5.15 (1H, m), 4.97 (2H, d, J=5.5 Hz),
4.82 (2H, s), 4.59 (1H, t, J=5.5 Hz), 4.22 (2H, br, s), 3.96-3.86
(1H, m), 3.33 (1H, dd, J=2.7, 11.4 Hz), 3.19-3.13 (1H, m), 2.69
(1H, td, J=2.5, 12.4 Hz), 2.27 (1H, d, J=12.2 Hz), 1.89-1.55
(10H+H.sub.2O, m), 1.30-1.14 (2H, m).
Example 54: tert-Butyl
(2R)-4-{[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetyl]amino}piperidine-2-carboxylate
##STR00128##
[0640] tert-Butyl
(2R)-4-{[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetyl]amino}piperidine-2-carboxylate was synthesised as
shown in Scheme 29.
##STR00129##
Stage 1:
(4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetic acid
[0641] To a solution of Intermediate 1 (1.34 g, 3.8 mmol) in DME
(20 mL), was added Intermediate 10 (956 mg, 4.2 mmol) and 2N
Na.sub.2CO.sub.3 (4.8 mL, 9.6 mmol) and the solution was degassed
by bubbling nitrogen through the reaction mixture. Dichlorobis
(triphenylphosphine) palladium (II) (270 mg, 0.4 mmol) was added
and the reaction was stirred at 90.degree. C. under nitrogen for 18
hrs. The reaction mixture was concentrated in vacuo and the residue
was dissolved in water (100 mL) and washed with EtOAc (50 mL). The
aqueous was filtered and acidified to pH3 using 1N HCl. The
resulting solid was collected by filtration, washed with water and
dried in a vacuum oven to give the title compound as a beige solid
(400 mg, 26%).
[0642] LCMS: m/z 393/395 [M+H].sup.+.
Stage 2: Di-tert-butyl
(2R)-4-{[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetyl]amino}piperidine-1,2-dicarboxylate
[0643]
(4-{5-Amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazol-1-
-yl)acetic acid (100 mg, 0.26 mmol) was dissolved in DMF (5 mL) and
EDC (59 mg, 0.31 mmol) and HOBt (41 mg, 0.31 mmol) were added. The
reaction was stirred for 20 mins after which time Intermediate 25
(92 mg, 0.31 mmol) was added. The reaction was allowed to stir for
18 hours at RT. The reaction mixture was diluted with EtOAc (25 mL)
and washed with 2N NaOH (10 mL), water (10 mL), brine (10 mL),
dried over MgSO.sub.4, filtered and concentrated in vacuo. The
crude material was purified by column chromatography (5%
MeOH/EtOAc) to give the title compound as a yellow solid (150 mg,
87%).
[0644] LCMS: m/z 675/677 [M+H].sup.+.
Stage 3: tert-Butyl
(2R)-4-{[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetyl]amino}piperidine-2-carboxylate
[0645] Di-tert-butyl
(2R)-4-{[(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazo-
l-1-yl)acetyl]amino}piperidine-1,2-dicarboxylate (150 mg, 0.22
mmol) was dissolved in dioxane (10 mL) and the reaction was cooled
to 0.degree. C. 4N HCl in dioxane (20 mL) was added. The reaction
mixture was stirred at 0.degree. C. for 2 hrs until completion. The
reaction was poured onto 2N NaOH (50 mL) and extracted with EtOAc
(3.times.25 mL). The organics were washed with brine (25 mL), dried
over MgSO.sub.4, filtered and concentrated in vacuo before
purification by preparative HPLC to give the title compound as an
off-white solid (10 mg, 8%).
[0646] LCMS: purity 96%, m/z 575/577 [M+H].sup.+.
[0647] .sup.1H NMR (300 MHz, CDCl.sub.3) 8.03 (1H, s), 7.92 (1H,
s), 7.69 (1H, s), 7.38-7.35 (2H, m), 7.24-7.19 (1H, m), 6.36 (1H,
d, J=7.9 Hz), 4.98 (2H, d, J=5.6 Hz), 4.83 (2H, s), 4.55 (1H, t,
J=5.4 Hz), 4.18 (2H, br s), 3.98-3.87 (1H, m), 3.28 (1H, dd, J=2.5,
11.3 Hz), 3.16 (1H, d, J=12.6 Hz), 2.73-2.64 (1H, m), 2.26 (1H, d,
J=12.1 Hz), 1.92-1.83 (2H, m), 1.44 (9H, s), 1.29-1.12 (2H, m).
Example 55: Cyclopentyl
(2R)-4-{[2-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyra-
zol-1-yl)ethyl]amino}piperidine-2-carboxylate di-formate
##STR00130##
[0649] Cyclopentyl
(2R)-4-{[2-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyra-
zol-1-yl)ethyl]amino}piperidine-2-carboxylate was synthesised as
shown in Scheme 30.
##STR00131##
Stage 1. Di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[1-(2-hydroxyethyl)-
-1H-pyrazol-4-yl]pyrazin-2-yl}imidodicarbonate
[0650] To a solution of Intermediate 4 (951 mg, 1.46 mmol) in DME
(20 mL), was added Intermediate 11 (524 mg, 2.20 mmol) and 2N
Na.sub.2CO.sub.3 (1.8 mL, 3.65 mmol) and the solution was degassed
by bubbling nitrogen through the reaction mixture for 20 mins.
Dichlorobis (triphenylphosphine) palladium (11) (270 mg, 0.4 mmol)
was added and the reaction was stirred at 80.degree. C. under
nitrogen for 5 hrs. The reaction mixture was cooled to RT and
diluted was EtOAc (100 mL). The reaction was filtered through
Celite.RTM. and the filter cake was washed with EtOAc (50 mL). The
filtrated was washed with brine (100 mL), dried over MgSO.sub.4,
filtered, concentrated in vacuo and purified by automated column
chromatography using EtOAc in heptane (gradient 10-100%) to give
the title compound as an off white solid (920 mg, 93%).
[0651] LCMS: m/z 579/581/583 [M-Boc].sup.+.
Stage 2. Di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[1-(2-oxoethyl)-1H--
pyrazol-4-yl]pyrazin-2-yl}imidodicarbonate
[0652] To a solution of di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[1-(2-hydroxyethyl)-
-1H-pyrazol-4-yl]pyrazin-2-yl}imidodicarbonate (900 mg, 1.32 mmol)
in DCM (14 mL) at 0.degree. C. was added Dess-Martin periodinone
(618 mg, 1.46 mmol). The reaction mixture was stirred with warming
to RT under nitrogen for 4 hrs. A further 0.5 eq of Dess-Martin
periodinone (300 mg) was added and the reaction stirred for 30
mins. The reaction was diluted with DCM (30 mL), sat NaHCO.sub.3
(50 mL) and sat sodium thiosulfate solution (50 mL) were added and
vigorously stirred for 1 hr. The aqueous was separated and washed
with DCM (100 mL). The combined organics were washed with sat
NaHCO.sub.3 and sat sodium thiosulfate solution (1:1 mixture, 200
mL), brine (200 mL), dried over MgSO.sub.4, filtered and
concentrated in vacuo to give the title compound as a yellow solid
(830 mg, 93%). The material was taken forward unpurified.
[0653] LCMS: m/z 577/579 [M-Boc]+.
Stage 3. 1-tert-Butyl 2-cyclopentyl
(2R)-4-{[(4-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)(2-
,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazol-1-yl)acetyl]amino}piperid-
ine-1,2-dicarboxylate
[0654] Di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[1-(2-oxoethyl)-1H--
pyrazol-4-yl]pyrazin-2-yl}imidodicarbonate (100 mg, 0.15 mmol),
Intermediate 22 (69 mg, 0.22 mmol) and acetic acid (168 .mu.L, 0.94
mmol) were dissolved in DCE (5 mL) and stirred at RT for 2 hrs.
STAB (93 mg, 0.44 mmol) was added and the reaction was stirred for
a further 2 hrs until completion. The reaction was diluted with DCM
(20 mL) and washed with sat NaHCO.sub.3 (20 mL). The aqueous layer
was separated and further extracted with DCM (20 mL). The combined
organics were dried over MgSO.sub.4, filtered, concentrated in
vacuo and purified by automated column chromatography using EtOAc
in heptane (gradient 0-100%) to give the title compound as a
colourless oil (63 mg, 44%).
[0655] LCMS: m/z 973/975/977 [M+H].sup.+.
Stage 4. Cyclopentyl
(2R)-4-{[2-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyra-
zol-1-yl)ethyl]amino}piperidine-2-carboxylate
[0656] To a solution of 1-tert-butyl 2-cyclopentyl
(2R)-4-{[(4-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)(2-
,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazol-1-yl)acetyl]amino}piperid-
ine-1,2-dicarboxylate (63 mg, 0.065 mmol) in dioxane (2 ml) was
added 4M HCl in dioxane (2 mL) and the reaction was stirred at RT
for 17 hrs. The reaction was concentrated in vacuo and the
resulting residue was purified by preparative HPLC to give the
di-formate salt of the title compound as a brown solid (11.1 mg,
30%).
[0657] LCMS purity 96%, m/z 573/575/577 [M+H].sup.+.
[0658] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 8.57 (2H, br,
s), 8.04 (1H, s), 7.92 (1H, s), 7.39-7.57 (3H, m), 7.32 (1H, s),
5.18-5.35 (1H, m), 4.96 (2H, s), 4.28-4.42 (2H, m), 3.77 (1H, d,
J=11.1 Hz), 3.21-3.30 (2H, m), 2.82-3.05 (2H, m), 2.35-2.46 (1H,
m), 2.03-2.14 (1H, m), 1.83-1.97 (2H, m), 1.74 (9H, br, s)
Example 56: Cyclopentyl
(2S)-4-{[2-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyra-
zol-1-yl)ethyl]amino}piperidine-2-carboxylate di-formate
##STR00132##
[0660] Cyclopentyl
(2S)-4-{[2-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyra-
zol-1-yl)ethyl]amino}piperidine-2-carboxylate was prepared in a
similar manner to Example 55 using Intermediate 23 at Stage 3 of
the route shown in Scheme 30.
[0661] LCMS: purity 98%, m/z 573/575/577 [M+H].sup.+.
[0662] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 8.19-8.87
(2H, m), 8.04 (1H, s), 7.92 (1H, s), 7.37-7.56 (3H, m), 7.22-7.36
(1H, m), 5.17-5.33 (1H, m), 4.96 (2H, s), 4.27-4.42 (2H, m), 3.78
(1H, d, J=11.5 Hz), 3.23-3.29 (2H, m), 2.80-3.07 (2H, m), 2.33-2.52
(1H, m), 2.02-2.15 (1H, m), 1.83-2.00 (2H, m), 1.74 (9H, d, J=3.8
Hz).
Example 57: Cyclopentyl
N-[2-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazol-1--
yl)ethyl]-L-leucinate
##STR00133##
[0664] Cyclopentyl
N-[2-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazol-1--
yl)ethyl]-L-leucinate was prepared in a similar manner to Example
55 using the free base of Intermediate 32 at Stage 3 and DCM as the
solvent in Stage 4 of the route shown in Scheme 30. The final
product was purified by preparative HPLC.
[0665] LCMS: purity 99%, m/z 560/562/564 [M+H].sup.+.
[0666] .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. ppm: 8.06 (1H, s),
7.90 (1H, s), 7.40-7.52 (3H, m), 7.34 (1H, s), 5.10-5.20 (1H, m),
4.95 (2H, s), 4.26 (2H, t, J=5.8 Hz), 3.01-3.19 (2H, m), 2.83-2.96
(1H, m), 1.53-1.90 (9H, m), 1.38-1.47 (2H, m), 0.89 (6H, d, J=2.6
Hz).
Example 58: tert-Butyl
N-[2-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazol-1--
yl)ethyl]-L-leucinate
##STR00134##
[0668] tert-Butyl
N-[2-(4-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}-1H-pyrazol-1--
yl)ethyl]-L-leucinate was prepared in a similar manner to Example
55 using L-leucine tert-butyl ester hydrochloride at Stage 3 and
DCM as the solvent in Stage 4 of the route shown in Scheme 30. The
final product was purified by preparative HPLC.
[0669] LCMS: purity 95%, m/z 548/550/552 (M+H).sup.+.
[0670] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. ppm: 8.06 (1H, s),
7.90 (1H, s), 7.40-7.52 (3H, m), 7.34 (1H, s), 5.10-5.20 (1H, m),
4.95 (2H, s), 4.26 (2H, t, J=5.8 Hz), 3.01-3.19 (2H, m), 2.83-2.96
(1H, m), 1.53-1.90 (9H, m), 1.38-1.47 (2H, m), 0.89 (6H, d, J=2.8
Hz).
Example 59 Cyclopentyl
4-[(5-{5-amino-6-[(2,6-dichlorobenzyl)amino]pyrazin-2-yl}pyridin-2-yl)met-
hyl]piperazine-2-carboxylate
##STR00135##
[0672] 1-ter-Butyl 2-cyclopentyl
4-[(5-{5-[bis(ter-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)(2,6-dich-
lorobenzyl)amino]pyrazin-2-yl}pyridin-2-yl)methyl]piperazine-1,2-dicarboxy-
late (0.078 g, 0.081 mmol) was stirred with TFA (3 mL) in
1,2-dicarboxylate (0.078 g, 0.081 mmol) was stirred with TFA (3 mL)
in dichloromethane (3 mL) for 1 h at room temperature. The reaction
was concentrated under reduced pressure and the residue was
purified by HPLC to give the title compound (28 mg).
[0673] LCMS purity 100%, m/z 556/558 [M+H].sup.+.
[0674] 1H NMR (300 MHz, CDCl.sub.3) 9.13 (1H, d, J=1.7 Hz), 8.23
(1H, dd, J=2.3, 8.1 Hz), 7.93 (1H, s), 7.47 (1H, d, J=8.1 Hz),
7.37-7.34 (2H, m), 7.20 (1H, dd, J=7.4, 8.7 Hz), 5.25-5.19 (1H, m),
5.03 (2H, d, J=5.5 Hz), 4.60 (1H, t, J=5.5 Hz), 4.36 (2H, s), 3.77
(1H, d, J=13.9 Hz), 3.67 (1H, d, J=13.9 Hz), 3.58 (1H, dd, J=3.2,
8.1 Hz), 3.13-3.06 (1H, m), 2.99-2.86 (2H, m), 2.69-2.63 (1H, m),
2.53-2.47 (1H, m), 2.39-2.31 (1H, m), 1.89-1.54 (9H, m).
[0675] The 1-tert-butyl 2-cyclopentyl
4-[(5-{5-[bis(tert-butoxycarbonyl)amino]-6-[(tert-butoxycarbonyl)
(2,6-dichlorobenzyl)amino]pyrazin-2-yl}pyridin-2-yl)methyl]piperazine-1,2-
-dicarboxylate used in the above process was prepared by the
reaction of di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-difluorobenzyl)amino]-5-(6-formylpyridin-3--
yl)pyrazin-2-yl}imidodicarbonate (100 mg, 0.15 mmol) with
Intermediate 28 (49 mg, 0.16 mmol) in DCE (5 mL) at room
temperature for 1.5 h. Sodium triacetoxyborohydride (63 mg, 0.30
mmol) was then added and stirring was continued at room temperature
for 2 h. Saturated aqueous Na.sub.2CO.sub.3 (20 mL) was added and
vigorous stirring continued for 20 min. The reaction was extracted
with dichloromethane (3.times.20 mL), dried (MgSO.sub.4) and
concentrated under reduced pressure. The residual yellow oil was
purified by column chromatography (silica gel; 50% EtOAc in
heptanes) to give the desired product (78 mg) as a colourless
oil.
[0676] LCMS: m/z 956/958 [M+H].sup.+.
[0677] The di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-difluorobenzyl)amino]-5-(6-formylpyridin-3--
yl)pyrazin-2-yl}imidodicarbonate used above was prepared by
dissolving di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[6-(hydroxymethyl)p-
yridin-3-yl]pyrazin-2-yl}imidodicarbonate (100 mg, 0.15 mmol) in
dichloromethane (5 mL) and adding manganese dioxide (257 mg, 3
mmol). The reaction was stirred at room temperature for 1 h and
then filtered through Celite.RTM.. The filter pad was washed with
dichloromethane (2.times.10 mL) and the combined filtrates
concentrated under reduced pressure to give an orange oil (100 mg)
which was used without purification.
[0678] The di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[6-(hydroxymethyl)p-
yridin-3-yl]pyrazin-2-yl}imidodicarbonate used above was prepared
by stirring di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[6-({[tert-butyl(di-
methyl)silyl}oxy}methyl)pyridin-3-yl]pyrazin-2-yl}imidodicarbonate
(1.22 g, 1.5 mmol) with tetrabutylammonium fluoride (3 mL, 3 mmol
1N in THF) in THF (20 mL) at room temperature for 17 h. The
reaction was diluted with EtOAc (50 mL) and washed with water
(2.times.50 mL) and brine (1.times.50 mL). The combined organic
layers were dried (MgSO.sub.4) and concentrated under reduced
pressure. The resulting pale yellow foam was purified by column
chromatography (silica gel; 80% EtOAc in heptanes) to give the
desired material (809 mg) as a colourless oil which solidified to a
white solid on standing. LCMS: m/z 676/678 [M+H].sup.+.
[0679] The di-tert-butyl
{3-[(tert-butoxycarbonyl)(2,6-dichlorobenzyl)amino]-5-[6-({[tert-butyl(di-
methyl)silyl}oxy}methyl)pyridin-3-yl]pyrazin-2-yl}imidodicarbonate
used above was prepared from Intermediate 4 (1.0 g, 1.5 mmol) and
2-({[tert-butyl(dimethyl)silyl]oxy}methyl)-5-(4,4,5,5-(4,4,5,5-tetramethy-
l-1,3,2-dioxaborolan-2-yl)pyridine (0.81 g, 2.3 mmol) which were
added to DME (20 mL) and 1.9 mL 2M Na.sub.2CO.sub.3 and the
resulting mixture was then degassed with N.sub.2 for 20 min.
Dichlorobis(triphenylphosphine)palladium(II) (108 mg, 0.2 mmol) was
added to the reaction mixture which was heated at 80.degree. C. for
16 h. The reaction was cooled and diluted with EtOAc (50 mL) and
filtered through Celite.RTM.. The filter pad was washed with EtOAc
(50 mL) and the combined filtrate was washed with brine (1.times.50
mL), dried MgSO.sub.4, and concentrated under reduced pressure. The
resulting residue was purified by column chromatography (silica
gel, 20% EtOAc in heptane) to give the desired material (1.22
g).
[0680] The
2-({[tert-butyl(dimethyl)silyl]oxy}methyl)-5-(4,4,5,5-tetrameth-
yl-1,3,2-dioxaborolan-2-yl)pyridine used in the above process was
prepared from
5-bromo-2-({[tert-butyl(dimethyl)silyl]oxy}methyl)pyridine (1.8 g
6.0 mmol), bis(pinacolato)diboron (1.81 g, 7.1 mmol) and
PdCl.sub.2(dppf).sub.2 (218 mg, 0.3 mmol) in a manner similar to
Intermediate 8 to give the desired product (1.1 g) as a pale green
oil. LCMS: m/z 268 [M+H].sup.+.
[0681] The
5-bromo-2-({[tert-butyl(dimethyl)silyl]oxy}methyl)pyridine used in
the above process was prepared from 5-bromo-2-hydroxymethylpyridine
(2.5 g, 13 mmol) which was dissolved in DMF (25 mL) and imidazole
(1.81 g, 27 mmol) and tert-butyldimethylsilyl chloride (2.20 g, 15
mmol) were added. The reaction mixture was stirred at room
temperature for 50 min and then poured into water (50 mL). This was
extracted with diethyl ether (3.times.50 mL) and the combined
organic layers were washed with brine (1.times.50 mL), dried
(MgSO.sub.4) and concentrated under reduced pressure. The resulting
residue was purified by column chromatography (silica gel; 5% EtOAc
in heptanes) to give the desired material (3.75 g) as a colourless
oil.
[0682] LCMS: m/z 302/304 [M+H].sup.+.
Measurement of Biological Activities
CSF1R (FMS) Kinase Activity
[0683] The ability of compounds to inhibit CSF1R (FMS) kinase
activity was measured in an assay performed by Invitrogen (Paisley,
UK). The Z'-LYTE.TM. biochemical assay employs a
fluorescence-based, coupled-enzyme format and is based on the
differential sensitivity of phosphorylated and non-phosphorylated
peptides to proteolytic cleavage. The peptide substrate is labelled
with two fluorophores-one at each end--that make up a FRET pair. In
the primary reaction, the kinase transfers the gamma-phosphate of
ATP to a single serine or threonine residue in a synthetic
FRET-peptide. In the secondary reaction, a site-specific protease
recognizes and cleaves non-phosphorylated FRET-peptides.
Phosphorylation of FRET-peptides suppresses cleavage by the
Development Reagent. Cleavage disrupts FRET between the donor (i.e.
coumarin) and acceptor (i.e. fluorescein) fluorophores on the
FRET-peptide, whereas uncleaved, phosphorylated FRET-peptides
maintain FRET. A radiometric method, which calculates the ratio
(the Emission Ratio) of donor emission to acceptor emission after
excitation of the donor fluorophore at 400 nm, is used to
quantitate reaction progress.
[0684] The final 10 .mu.L Kinase Reaction consists of 0.12-12.3 ng
CSF1R (FMS), 2 .mu.M Tyr 01 Peptide and ATP in 50 mM HEPES pH 7.5,
0.01% BRIJ-35, 10 mM MgCl.sub.2, 1 mM EGTA. The assay is performed
at an ATP concentration at, or close to, the Km. After the 60
minute Kinase Reaction incubation at room temperature, 5 .mu.L of a
1:256 dilution of Development Reagent is added. The assay plate is
incubated for a further 60 minutes at room temperature and read on
a fluorescence plate reader.
[0685] Duplicate data points are generated from a 1/3 log dilution
series of a stock solution of test compound in DMSO. Nine dilutions
steps are made from a top concentration of 10 mM, and a `no
compound` blank is included. Data is collected and analysed using
XLfit software from IDBS. The dose response curve is curve fitted
to model number 205 (sigmoidal dose-response model). From the curve
generated, the concentration giving 50% inhibition is determined
and reported.
Macrophage Colony-Stimulating Factor (MCSF)-Stimulation of Human
Whole Blood
[0686] Whole blood was taken by venous puncture using heparinised
vacutainers (Becton Dickinson) and diluted in an equal volume of
RPM11640 tissue culture media (Sigma). 100 .mu.l was plated in
V-bottomed 96 well tissue culture treated plates. 2 hours after the
addition of the inhibitor in 100 .mu.l of RPMI1640 media, the blood
was stimulated with MCSF (Peprotech: #300-25-10 uG) at a final
concentration of 50 ng/ml and incubated at 37.degree. C. in 5%
CO.sub.2 for 22 hrs. MCP-1 levels were measured from cell-free
supernatants by sandwich ELISA developed from MCP-1/CCL2 Duoset
(R&D Systems #DY279). Additional reagents required for ELISA
were purchased from Sigma, (BSA: A3059, 10.times.PBS: D1408,
Tween20: P5927) R&D Systems, (colour reagent: #DY999 and stop
solution: #DY994) and Costar (96-well EIA plates: #2592). Assays
were carried out following the manufacturer's instructions with the
following modifications: Cell-free supernatants were diluted 30:70
in reagent diluent before assay. All incubations except capture
antibody were carried out with shaking at 550 rpm. 3 washes were
carried out between steps, and incubation times were blocking 1.5
hr, sample/standard and detection antibody incubations 2 hr, HRP
incubation 30 min, colour development 20 min.
[0687] IC50 values were allocated to one of five ranges as
follows:
TABLE-US-00001 Inhibitor activity Inhibitor activity Example versus
CSF-1 versus MCP-1 1 A C 2 B C 3 B C 4 B NT 5 A D 6 B C 7 A B 8 A B
9 C A 10 D A 11 C A 12 E NT 13 D NT 14 B C 15 E NT 16 D NT 17 A NT
18 B B 19 A A 20 D A 21 A C 22 A C 23 C C 24 C NT 25 B C 26 A C 27
B A 28 B A 29 NT D 30 B B 31 B A 32 B D 33 B D 34 A B 35 A C 36 C B
37 C A 38 E E 39 E E 40 E D 41 D NT 42 E C 43 E E 44 E E 45 D D 46
A C 47 A C 48 D C 49 D D 50 A B 51 A B 52 NT B 53 B NT 54 B NT 55 B
B 56 B C 57 D D 58 E E 59 B NT Range A: IC50 < 100 nM Range B:
100 nM < IC50 < 300 nM Range C: 300 nM < IC50 < 1000 nM
Range D: 1000 nM < IC50 < 5000 nM Range E: IC50 > 5000 nM
NT = not tested
* * * * *