U.S. patent application number 12/171591 was filed with the patent office on 2008-11-20 for pyrimidine derivatives useful as inhibitors of pkc-theta.
Invention is credited to Antonio J.M. BARBOSA, Steven Richard BRUNETTE, Eugene Richard HICKEY, Jin Mi KIM, Michael David LAWLOR, Rene Marc LEMIEUX, Bryan MCKIBBEN, Matt Aaron TSCHANTZ, Hui YU.
Application Number | 20080287410 12/171591 |
Document ID | / |
Family ID | 34982334 |
Filed Date | 2008-11-20 |
United States Patent
Application |
20080287410 |
Kind Code |
A1 |
BARBOSA; Antonio J.M. ; et
al. |
November 20, 2008 |
PYRIMIDINE DERIVATIVES USEFUL AS INHIBITORS OF PKC-THETA
Abstract
Disclosed are novel compounds of formula (I): ##STR00001##
wherein X, Y, R.sub.1, R.sub.2 and R.sub.3 are as defined herein,
which are useful as inhibitors of PKC-theta and are thus useful for
treating a variety of diseases and disorders that are mediated or
sustained through the activity of PKC-theta, including
immunological disorders and type II diabetes. This invention also
relates to pharmaceutical compositions comprising these compounds,
methods of using these compounds in the treatment of various
diseases and disorders, processes for preparing these compounds and
intermediates useful in these processes.
Inventors: |
BARBOSA; Antonio J.M.;
(Middlebury, CT) ; BRUNETTE; Steven Richard; (New
Milford, CT) ; HICKEY; Eugene Richard; (Danbury,
CT) ; KIM; Jin Mi; (Sandy Hook, CT) ; LAWLOR;
Michael David; (Seymour, CT) ; LEMIEUX; Rene
Marc; (Plantsville, CT) ; MCKIBBEN; Bryan;
(New Milford, CT) ; TSCHANTZ; Matt Aaron; (Newton,
CT) ; YU; Hui; (New Milford, CT) |
Correspondence
Address: |
MICHAEL P. MORRIS;BOEHRINGER INGELHEIM USA CORPORATION
900 RIDGEBURY RD, P O BOX 368
RIDGEFIELD
CT
06877-0368
US
|
Family ID: |
34982334 |
Appl. No.: |
12/171591 |
Filed: |
July 11, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11175476 |
Jul 6, 2005 |
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12171591 |
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60586289 |
Jul 8, 2004 |
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Current U.S.
Class: |
514/210.2 ;
514/275 |
Current CPC
Class: |
C07D 403/12 20130101;
A61P 17/00 20180101; A61P 29/00 20180101; A61P 3/10 20180101; A61P
37/06 20180101; A61P 1/04 20180101; C07D 239/48 20130101; A61P
37/00 20180101; A61P 37/08 20180101; C07D 401/12 20130101; A61P
43/00 20180101; A61P 17/06 20180101 |
Class at
Publication: |
514/210.2 ;
514/275 |
International
Class: |
A61K 31/505 20060101
A61K031/505; A61K 31/397 20060101 A61K031/397; A61P 37/00 20060101
A61P037/00 |
Claims
1.-11. (canceled)
12. A method of treating a disease or disorder that is that is
mediated or sustained through the activity of inhibiting PKC-theta
in a patient comprising administering to said patient a
therapeutically effect amount of a compound of the following
formula (I): ##STR00162## wherein: X is C.sub.1-6 alkyl wherein one
or two of the methylene units can be replaced by an oxygen or
sulfur atom, and wherein the C.sub.1-6alkyl group is optionally and
independently substituted with: (A) oxo, (B) C.sub.1-6alkyl which
is optionally substituted with one or more of the following groups:
(i) hydroxyl, (ii) C.sub.1-3alkyloxy, (iii) halogen, (C)
C.sub.1-6alkyloxy, (D) C.sub.1-6alkylthio, (E) aryl (F)
--COR.sub.6, wherein R.sub.6 is: (i) C.sub.1-6alkyl, (ii)
C.sub.1-6alkyloxy, (iii) --NR.sub.7R.sub.8, wherein R.sub.7 and
R.sub.8 are each independently selected from: (a) hydrogen, (b)
C.sub.1-6alkyl, (c) aryl, (d) heteroaryl, (iv) --OH, (G) halogen,
(H) --NR.sub.9R.sub.10, wherein R.sub.9 and R.sub.10 are each
independently selected from: (i) hydrogen, (ii) C.sub.1-6alkyl,
(iii) C.sub.1-6alkylcarbonyl, (iv) C.sub.1-6alkylsulfonyl, (v)
aryl, (vi) heteroaryl; Y is --NH--, --O-- or --S--; R.sub.1 is: (A)
aryl or heteroaryl, each optionally and independently substituted
with one or more of the following groups: (i) C.sub.1-6alkyl, which
is optionally substituted one or more of the following: (a)
halogen, (b) NH.sub.2 (ii) C.sub.1-6alkoxy, which is optionally
substituted with halogen, (iii) C.sub.1-6alkylthio, which is
optionally substituted with halogen, (iv) C.sub.1-6alkylsulfonyl,
(v) cyano, (vi) halogen, (vii) hydroxyl, (viii) nitro, (ix)
--COR.sub.11, wherein R.sub.1 is: (a) C.sub.1-6alkyl, (b)
C.sub.1-6alkyloxy, (c) --OH, (d) --NR.sub.12R.sub.13, wherein
R.sub.12 and R.sub.13 are each independently selected from: (I)
hydrogen, (II) C.sub.1-6alkyl, (III) aryl, (IV) heteroaryl, (x)
--NR.sub.14R.sub.15, wherein R.sub.14 and R.sub.15 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl, (c)
C.sub.1-6alkylcarbonyl, (d) C.sub.1-6alkylsulfonyl, or wherein
R.sub.14 and R.sub.15 together constitute an alkylene bridge which
together with the nitrogen atom between them forms a five to
seven-membered ring, (xi) arylthio, arylsulfonyl or aryloxy, each
optionally and independently substituted with one or more of the
following groups: (a) C.sub.1-6alkyl, (b) C.sub.1-6alkoxy, (c)
C.sub.1-6alkylthio, (d) C.sub.1-6alkylsulfonyl, (e) cyano, (f)
halogen, (g) nitro, (h) --NR.sub.16R.sub.17, wherein R.sub.16 and
R.sub.17 are each independently selected from: (I) hydrogen, (II)
C.sub.1-6alkyl, (III) C.sub.1-6alkylcarbonyl, (IV)
C.sub.1-6alkylsulfonyl, (B) C.sub.3-6cycloalkyl which is optionally
and independently substituted with one or more of the following
groups: (i) C.sub.1-6alkyl, which is optionally substituted with
halogen, (ii) C.sub.1-6alkoxy, which is optionally substituted with
halogen, (iii) C.sub.1-6alkylthio, which is optionally substituted
with halogen, (iv) C.sub.1-6alkylsulfonyl, (v) halogen, (vi)
hydroxyl, (vii) --NR.sub.18R.sub.19, wherein R.sub.18 and R.sub.19
are each independently selected from: (a) hydrogen, (b)
C.sub.1-6alkyl, (c) C.sub.1-6alkylcarbonyl, (d)
C.sub.1-6alkylsulfonyl, (viii) --COR.sub.20, wherein R.sub.20 is:
(a) C.sub.1-6alkyl, (b) C.sub.1-6alkyloxy, (c) --OH, (d)
--NR.sub.21R.sub.22, wherein R.sub.21 and R.sub.22 are each
independently selected from: (I) hydrogen, (II) C.sub.1-6alkyl, (C)
--COR.sub.23, wherein R.sub.23 is: (i) C.sub.1-6alkyloxy, (ii)
--NR.sub.24R.sub.25, wherein R.sub.24 and R.sub.25 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl, or
wherein R.sub.24 and R.sub.25 together constitute an alkylene
bridge which together with the nitrogen atom between them forms a
four to six-membered ring, wherein one of the methylene groups is
optionally replaced by an oxygen, sulfur or NH group, and which
ring is optionally and independently substituted with one or more
of the following groups: (a) C.sub.1-6alkyloxy, (b) C.sub.1-6alkyl,
which is optionally substituted with halogen, (c) hydroxyl, (d)
halogen, (e) --COR.sub.26, wherein R.sub.26 is: (I)
C.sub.1-6alkyloxy, (II) --NR.sub.27R.sub.28, wherein R.sub.27 and
R.sub.28 are each independently selected from: a. hydrogen, b.
C.sub.1-6alkyl, c. aryl, d. heteroaryl, (D) is: ##STR00163## which
is optionally substituted with halogen; (E) is selected from the
following: ##STR00164## R.sub.2 is selected from the following
groups: (A) CF.sub.3, (B) cyano, (C) halogen, (D) nitro, (E)
C.sub.1-6alkylalkynyl, (F) arylalkynyl which is optionally
substituted with one or more of the following groups: (i) halogen,
(ii) C.sub.1-6alkyl, which is optionally substituted with halogen,
R.sub.3 is selected from the following groups: ##STR00165## R.sub.4
and R.sub.5 are each independently selected from: (A) hydrogen, (B)
C.sub.1-6alkyl, optionally and independently substituted with one
or more of the following groups: (i) oxo, (ii) C.sub.3-6cycloalkyl,
(iii) C.sub.1-6alkyloxy, (iv) C.sub.1-6alkylthio, (v) --COR.sub.29,
wherein R.sub.29 is: (a) C.sub.1-6alkyl, (b) C.sub.1-6alkyloxy, (c)
--OH, (vi) --CONR.sub.30R.sub.31, wherein R.sub.30 and R.sub.31 are
each independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl,
(c) aryl, (d) heteroaryl, (vii) halogen, (viii) hydroxyl, (ix)
--NR.sub.32R.sub.33, wherein R.sub.32 and R.sub.33 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl, (c)
C.sub.1-6alkylcarbonyl, (d) C.sub.1-6alkylsulfonyl, (e) aryl, (f)
heteroaryl, (x) aryl or heteroaryl, each optionally and
independently substituted with one or more of the following groups:
(a) C.sub.1-6alkyl, which is optionally substituted with halogen,
(b) C.sub.1-6alkoxy, which is optionally substituted with halogen,
(c) C.sub.1-6alkylthio, which is optionally substituted with
halogen, (d) C.sub.1-6alkylsulfonyl, (e) cyano, (f) halogen, (g)
nitro, (h) --NR.sub.34R.sub.35, wherein R.sub.34 and R.sub.35 are
each independently selected from: (I) hydrogen, (II)
C.sub.1-6alkyl, (III) C.sub.1-6alkylcarbonyl, (IV)
C.sub.1-6alkylsulfonyl, (i) --COR.sub.36, wherein R.sub.36 is: (I)
C.sub.1-6alkyl, (II) C.sub.1-6alkyloxy, (III) --OH, (j)
--CONR.sub.37R.sub.38, wherein R.sub.37 and R.sub.38 are each
independently selected from: (I) hydrogen, (II) C.sub.1-6alkyl,
(III) aryl, (IV) heteroaryl, (C) C.sub.1-6alkylsulfonyl, (D)
arylsulfonyl, (E) aryl-C.sub.1-6alkylsulfonyl, (F)
heteroarylsulfonyl, (G) C.sub.1-6alkylcarbonyl, (H) arylcarbonyl,
(I) aryl-C.sub.1-6alkylcarbonyl, (J) heteroarylcarbonyl, (K)
C.sub.1-6alkylaminocarbonyl, or (L) heteroaryl, or wherein R.sub.4
and R.sub.5 together constitute an alkylene bridge which together
with the nitrogen atom between them forms a four- to seven-membered
ring, wherein one of the methylene groups is optionally replaced by
an oxygen, sulfur or NH group, and which ring is optionally and
independently substituted with one or more of the following groups:
(A) C.sub.1-6alkyl, which is optionally substituted with halogen,
(B) C.sub.1-6alkoxy, which is optionally substituted with halogen,
(C) C.sub.1-6alkylthio, which is optionally substituted with
halogen, (D) C.sub.1-6alkylsulfonyl, (E) halogen, (F)
--NR.sub.39R.sub.40, wherein R.sub.39 and R.sub.40 are each
independently selected from: (i) hydrogen, (ii) C.sub.1-6alkyl,
(iii) C.sub.1-6alkylcarbonyl, (iv) C.sub.1-6alkylsulfonyl, (v)
arylcarbonyl, (vi) arylsulfonyl (vii) heteroarylcarbonyl, (viii)
heteroarylsulfonyl, (G) --COR.sub.41, wherein R.sub.1 is: (i)
C.sub.1-6alkyl, (ii) C.sub.1-6alkyloxy, (iii) --NR.sub.42R.sub.43,
wherein R.sub.42 and R.sub.43 are each independently selected from:
(a) hydrogen, (b) C.sub.1-6alkyl, (c) aryl, (d) heteroaryl, (iv)
hydroxyl, (H) --OR.sub.44, wherein R.sub.44 is selected from: (i)
hydrogen, (ii) C.sub.1-6alkylcarbonyl, (iii) C.sub.1-6alkylsulfonyl
(I) oxo, or wherein NR.sub.4R.sub.5 constitutes a 5-membered
heteroaryl ring containing a total of 2 nitrogen hetero atoms in
the ring; or a tautomer or pharmaceutically acceptable salt thereof
or a corresponding compound having at least one amine group
protected by an amino-protecting group.
13. A method of claim 12, wherein the compound administered is of
formula (I) wherein: X is C.sub.1-3alkyl, optionally substituted
with oxo, Y is --NH-- or --O--; R.sub.1 is: (A) aryl or heteroaryl,
each optionally and independently substituted with one or more of
the following groups: (i) C.sub.1-3alkyl, which is optionally and
independently substituted with fluorine, (ii) C.sub.1-3alkoxy,
which is optionally and independently substituted with fluorine,
(iii) C.sub.1-3alkylthio, which is optionally and independently
substituted with fluorine, (iv) arylthio, which is optionally
substituted with --NH.sub.2, (v) halogen, (vi) hydroxyl, (vii)
C.sub.1-3alkylsulfonyl, (viii) CONH.sub.2, (ix)
--NR.sub.14R.sub.15, wherein R.sub.14 and R.sub.15 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl (c)
C.sub.1-6alkylcarbonyl, (d) C.sub.1-6alkylsulfonyl, or wherein
R.sub.14 and R.sub.15 together constitute an alkylene bridge which
together with the nitrogen atom between them forms a five to
seven-membered ring, (B) cyclohexyl, which is optionally and
independently substituted with: (i) C.sub.1-3alkyl, (ii) hydroxyl;
(c) --COR.sub.23, wherein R.sub.23 is: (i) --NR.sub.24R.sub.25,
wherein R.sub.24 and R.sub.25 are each independently selected from:
(a) C.sub.1-6alkyl; or wherein R.sub.24 and R.sub.25 together
constitute a alkylene bridge which together with the nitrogen atom
between them forms a five to six-membered ring, wherein one of the
methylene groups is optionally replaced by an oxygen atom and which
ring is optionally and independently substituted with one or more
of the following groups: (ii) hydroxyl, (iii) --CONH.sub.2, (D) is:
##STR00166## which is optionally substituted with halogen; (E) is
selected from the following: ##STR00167## R.sub.2 is: (A) CF.sub.3,
(B) cyano, (C) halogen, or (D) nitro, R.sub.3 is selected from the
following: ##STR00168## R.sub.4 and R.sub.5 are each independently
selected from: (A) hydrogen, (B) C.sub.1-6alkyl, which is
optionally and independently substituted with one or more of the
following groups: (i) oxo, (ii) C.sub.3-5cycloalkyl, (iii) aryl or
heteroaryl, each of which is optionally and independently
substituted with one or more of the following groups: (a)
C.sub.1-3alkyl which is optionally substituted with fluorine, (b)
--CO.sub.2H (c) halogen, (iv) NH.sub.2 (v) hydroxyl, (vi)
--CONH.sub.2, (vii) fluorine, (viii) NHCOCH.sub.3 or R.sub.4 and
R.sub.5 together constitute an alkylene bridge which together with
the nitrogen atom between them forms a four- to seven-membered
ring, wherein one of the methylene groups is optionally replaced by
an NH or oxygen atom and which ring is optionally and independently
substituted with one or more of the following groups: (A)
--CONH.sub.2, (B) NR.sub.39R.sub.40 wherein R.sub.39 and R.sub.40
are optionally and independently selected from: (i)
C.sub.1-5alkylcarbonyl (ii) C.sub.1-5alkylsulfonyl (iii)
arylcarbonyl (iv) arylsulfonyl (C) --OR.sub.4, wherein R.sub.4 is
selected from: (i) hydrogen, (ii) C.sub.1-5alkylcarbonyl (D) oxo,
or (E) fluorine or wherein NR.sub.4R.sub.5 constitute a 5-membered
heteroaryl ring containing a total of 2 nitrogen hetero atoms in
the ring, or a tautomer or pharmaceutically acceptable salt thereof
or a corresponding compound having at least one amine group
protected by an amino-protecting group.
14. A method of claim 12, wherein the compound administered is of
the formula (I) wherein: X is --CH.sub.2--, --CH.sub.2CH.sub.2-- or
--CH.sub.2CO--, Y is --NH-- or --O--; R.sub.1 is: (A) phenyl,
pyridyl, naphthyl, quinolinyl or benzothienyl, each of which is
optionally and independently substituted with one or more of the
following groups: (i) C.sub.1-3alkyl, which is optionally
substituted with fluorine, (ii) C.sub.1-3alkoxy, which is
optionally substituted with fluorine, (iii) methylthio, which is
optionally substituted with fluorine, (iv) arylthio, optionally
substituted with NH.sub.2, (v) F, Cl or Br, (vi) hydroxyl, (vii)
NH.sub.2 or N(CH.sub.3).sub.2, (viii) SO.sub.2CH.sub.3 (B)
cyclohexyl, optionally substituted with hydroxyl, R.sub.2 is: (A)
CF.sub.3, (B) cyano, (C) F, Cl, Br or (D) nitro, R.sub.3 is:
##STR00169## R.sub.4, R.sub.5 are each independently selected from:
(A) hydrogen, (B) C.sub.1-4alkyl, which is optionally and
independently substituted with one or more of the following groups:
(i) oxo, (ii) cyclopropyl, (iii) aryl or heteroaryl selected from
phenyl, pyridyl, pyrimidyl, pyrazolyl and oxazolyl, each of which
is optionally and independently substituted with one or more of the
following groups: (a) C.sub.1-6alkyl, (b) fluorine or chlorine,
(iv) --CONH.sub.2 (v) hydroxyl, (vi) fluorine, (vii) NHCOCH.sub.3
or R.sub.4 and R.sub.5 together constitute an alkylene bridge which
together with the nitrogen atom between them forms a four- to
seven-membered ring, wherein one of the methylene groups is
optionally replaced by an NH, and which ring is optionally and
independently substituted with one or more of the following groups:
(A) CONH.sub.2, (B) hydroxyl, (C) C.sub.1-5alkylcarbonyloxy, (D)
oxo, (E) fluorine, (F) NR.sub.39R.sub.40 wherein R.sub.39 and
R.sub.40 are optionally and independently selected from: (i)
hydrogen (ii) C.sub.1-5alkylcarbonyl (iii) C.sub.1-5alkylsulfonyl
(iv) phenylcarbonyl, or (v) phenylsulfonyl, or a tautomer or
pharmaceutically acceptable salt thereof or a corresponding
compound having at least one amine group protected by an
amino-protecting group.
15. A method of claim 12, wherein the compound administered is of
the formula (I) wherein: X is --CH.sub.2--, Y is --NH--, R.sub.1
is: (A) phenyl or pyridyl optionally and independently substituted
with one or more of the following groups: (i) methyl, (ii)
CF.sub.3, (iii) OCF.sub.3, (iv) OCF.sub.2H (v) OCH.sub.3, (vi)
OCH(CH.sub.3).sub.2 (vii) SCF.sub.3, (viii) arylthio substituted
with NH.sub.2, (ix) F or Cl, (x) N(CH.sub.3).sub.2, (xi)
OCH.sub.2CF.sub.3 (xii) SO.sub.2CH.sub.3 (B) naphthyl, R.sub.2 is:
(A) cyano, (B) Cl, or (C) nitro; R.sub.3 is: ##STR00170## R.sub.4
and R.sub.5 are each independently selected from: (A) hydrogen, (B)
C.sub.1-3alkyl, optionally substituted with one or more of the
following groups: (i) hydroxyl, (ii) pyridyl, (iii)
1-methyl-1H-pyrazole, (iv) 1,5-dimethyl-1H-pyrazole, (v)
--CONH.sub.2, or R.sub.4 and R.sub.5 together constitute an
alkylene bridge which together with the nitrogen atom between them
forms a four or five-membered ring, each optionally and
independently substituted with one or more of the following: (A)
hydroxyl, (B) amino (C) fluorine (D) oxo (E) --NHSO.sub.2CH.sub.3
(F) --NHCOCH.sub.3 or a tautomer or pharmaceutically acceptable
salt thereof or a corresponding compound having at least one amine
group protected by an amino-protecting group.
16. A method of claim 12, wherein the compound administered is of
the formula (I) wherein: X is --CH.sub.2--, Y is --NH--, R.sub.1
is: (A) phenyl, optionally and independently substituted with one
or more of the following groups: (i) methyl, (ii) CF.sub.3, (iii)
OCF.sub.3, (iv) OCH.sub.3, (v) SCF.sub.3, (vi) arylthio substituted
with NH.sub.2, (vii) F or Cl; (B) naphthyl, R.sub.2 is: (A) cyano,
(B) Cl, or (C) nitro; R.sub.3 is: ##STR00171## R.sub.4 and R.sub.5
are each independently selected from: (A) hydrogen, (B)
C.sub.1-2alkyl, optionally substituted with one or more of the
following groups: (i) hydroxyl, (ii) pyridyl, (iii)
1,5-dimethyl-1H-pyrazole, (iv) --CONH.sub.2, or R.sub.4 and R.sub.5
together constitute an alkylene bridge which together with the
nitrogen atom between them forms a four or five-membered ring, each
optionally and independently substituted with one or more hydroxyl
or oxo; or a tautomer or pharmaceutically acceptable salt thereof
or a corresponding compound having at least one amine group
protected by an amino-protecting group.
17. A method of treating a disease or disorder associated with the
activation of T cells in a patient comprising administering to said
patient a therapeutically effective amount of a compound of the
following formula (I): ##STR00172## wherein: X is C.sub.1-6 alkyl
wherein one or two of the methylene units can be replaced by an
oxygen or sulfur atom, and wherein the C.sub.1-6alkyl group is
optionally and independently substituted with: (A) oxo, (B)
C.sub.1-6alkyl which is optionally substituted with one or more of
the following groups: (i) hydroxyl, (ii) C.sub.1-3alkyloxy, (iii)
halogen, (C) C.sub.1-6alkyloxy, (D) C.sub.1-6alkylthio, (E) aryl
(F) --COR.sub.6, wherein R.sub.6 is: (i) C.sub.1-6alkyl, (ii)
C.sub.1-6alkyloxy, (iii) --NR.sub.7R.sub.8, wherein R.sub.7 and
R.sub.8 are each independently selected from: (a) hydrogen, (b)
C.sub.1-6alkyl, (c) aryl, (d) heteroaryl, (iv) --OH, (G) halogen,
(H) --NR.sub.9R.sub.10, wherein R.sub.9 and R.sub.10 are each
independently selected from: (i) hydrogen, (ii) C.sub.1-6alkyl,
(iii) C.sub.1-6alkylcarbonyl, (iv) C.sub.1-6alkylsulfonyl, (v)
aryl, (vi) heteroaryl; Y is --NH--, --O-- or --S--; R.sub.1 is: (A)
aryl or heteroaryl, each optionally and independently substituted
with one or more of the following groups: (i) C.sub.1-6alkyl, which
is optionally substituted one or more of the following: (a)
halogen, (b) NH.sub.2 (ii) C.sub.1-6alkoxy, which is optionally
substituted with halogen, (iii) C.sub.1-6alkylthio, which is
optionally substituted with halogen, (iv) C.sub.1-6alkylsulfonyl,
(v) cyano, (vi) halogen, (vii) hydroxyl, (viii) nitro, (ix)
--COR.sub.11, wherein R.sub.11 is: (a) C.sub.1-6alkyl, (b)
C.sub.1-6alkyloxy, (c) --OH, (d) --NR.sub.12R.sub.13, wherein
R.sub.12 and R.sub.13 are each independently selected from: (I)
hydrogen, (II) C.sub.1-6alkyl, (III) aryl, (IV) heteroaryl, (x)
--NR.sub.14R.sub.15, wherein R.sub.14 and R.sub.15 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl, (c)
C.sub.1-6alkylcarbonyl, (d) C.sub.1-6alkylsulfonyl, or wherein
R.sub.14 and R.sub.15 together constitute an alkylene bridge which
together with the nitrogen atom between them forms a five to
seven-membered ring, (xi) arylthio, arylsulfonyl or aryloxy, each
optionally and independently substituted with one or more of the
following groups: (a) C.sub.1-6alkyl, (b) C.sub.1-6alkoxy, (c)
C.sub.1-6alkylthio, (d) C.sub.1-6alkylsulfonyl, (e) cyano, (f)
halogen, (g) nitro, (h) --NR.sub.16R.sub.17, wherein R.sub.16 and
R.sub.17 are each independently selected from: (I) hydrogen, (II)
C.sub.1-6alkyl, (III) C.sub.1-6alkylcarbonyl, (IV)
C.sub.1-6alkylsulfonyl, (B) C.sub.3-6cycloalkyl which is optionally
and independently substituted with one or more of the following
groups: (i) C.sub.1-6alkyl, which is optionally substituted with
halogen, (ii) C.sub.1-6alkoxy, which is optionally substituted with
halogen, (iii) C.sub.1-6alkylthio, which is optionally substituted
with halogen, (iv) C.sub.1-6alkylsulfonyl, (v) halogen, (vi)
hydroxyl, (vii) --NR.sub.18R.sub.19, wherein R.sub.18 and R.sub.19
are each independently selected from: (a) hydrogen, (b)
C.sub.1-6alkyl, (c) C.sub.1-6alkylcarbonyl, (d)
C.sub.1-6alkylsulfonyl, (viii) --COR.sub.20, wherein R.sub.20 is:
(a) C.sub.1-6alkyl, (b) C.sub.1-6alkyloxy, (c) --OH, (d)
--NR.sub.21R.sub.22, wherein R.sub.21 and R.sub.22 are each
independently selected from: (I) hydrogen, (II) C.sub.1-6alkyl, (C)
--COR.sub.23, wherein R.sub.23 is: (i) C.sub.1-6alkyloxy, (ii)
--NR.sub.24R.sub.25, wherein R.sub.24 and R.sub.25 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl, or
wherein R.sub.24 and R.sub.25 together constitute an alkylene
bridge which together with the nitrogen atom between them forms a
four to six-membered ring, wherein one of the methylene groups is
optionally replaced by an oxygen, sulfur or NH group, and which
ring is optionally and independently substituted with one or more
of the following groups: (a) C.sub.1-6alkyloxy, (b) C.sub.1-6alkyl,
which is optionally substituted with halogen, (c) hydroxyl, (d)
halogen, (e) --COR.sub.26, wherein R.sub.26 is: (I)
C.sub.1-6alkyloxy, (II) --NR.sub.27R.sub.28, wherein R.sub.27 and
R.sub.28 are each independently selected from: a. hydrogen, b.
C.sub.1-6alkyl, c. aryl, d. heteroaryl, (D) is: ##STR00173## which
is optionally substituted with halogen; (E) is selected from the
following: ##STR00174## R.sub.2 is selected from the following
groups: (A) CF.sub.3, (B) cyano, (C) halogen, (D) nitro, (E)
C.sub.1-6alkylalkynyl, (F) arylalkynyl which is optionally
substituted with one or more of the following groups: (i) halogen,
(ii) C.sub.1-6alkyl, which is optionally substituted with halogen,
R.sub.3 is selected from the following groups: ##STR00175## R.sub.4
and R.sub.5 are each independently selected from: (A) hydrogen, (B)
C.sub.1-6alkyl, optionally and independently substituted with one
or more of the following groups: (i) oxo, (ii) C.sub.3-6cycloalkyl,
(iii) C.sub.1-6alkyloxy, (iv) C.sub.1-6alkylthio, (v) --COR.sub.29,
wherein R.sub.29 is: (a) C.sub.1-6alkyl, (b) C.sub.1-6alkyloxy, (c)
--OH, (vi) --CONR.sub.30R.sub.31, wherein R.sub.30 and R.sub.31 are
each independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl,
(c) aryl, (d) heteroaryl, (vii) halogen, (viii) hydroxyl, (ix)
--NR.sub.32R.sub.33, wherein R.sub.32 and R.sub.33 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl, (c)
C.sub.1-6alkylcarbonyl, (d) C.sub.1-6alkylsulfonyl, (e) aryl, (f)
heteroaryl, (x) aryl or heteroaryl, each optionally and
independently substituted with one or more of the following groups:
(a) C.sub.1-6alkyl, which is optionally substituted with halogen,
(b) C.sub.1-6alkoxy, which is optionally substituted with halogen,
(c) C.sub.1-6alkylthio, which is optionally substituted with
halogen, (d) C.sub.1-6alkylsulfonyl, (e) cyano, (f) halogen, (g)
nitro, (h) --NR.sub.34R.sub.35, wherein R.sub.34 and R.sub.35 are
each independently selected from: (I) hydrogen, (II)
C.sub.1-6alkyl, (III) C.sub.1-6alkylcarbonyl, (IV)
C.sub.1-6alkylsulfonyl, (i) --COR.sub.36, wherein R.sub.36 is: (I)
C.sub.1-6alkyl, (II) C.sub.1-6alkyloxy, (III) --OH, (j)
--CONR.sub.37R.sub.38, wherein R.sub.37 and R.sub.38 are each
independently selected from: (I) hydrogen, (II) C.sub.1-6alkyl,
(III) aryl, (IV) heteroaryl, (C) C.sub.1-6alkylsulfonyl, (D)
arylsulfonyl, (E) aryl-C.sub.1-6alkylsulfonyl, (F)
heteroarylsulfonyl, (G) C.sub.1-6alkylcarbonyl, (H) arylcarbonyl,
(I) aryl-C.sub.1-6alkylcarbonyl, (J) heteroarylcarbonyl, (K)
C.sub.1-6alkylaminocarbonyl, or (L) heteroaryl, or wherein R.sub.4
and R.sub.5 together constitute an alkylene bridge which together
with the nitrogen atom between them forms a four- to seven-membered
ring, wherein one of the methylene groups is optionally replaced by
an oxygen, sulfur or NH group, and which ring is optionally and
independently substituted with one or more of the following groups:
(A) C.sub.1-6alkyl, which is optionally substituted with halogen,
(B) C.sub.1-6alkoxy, which is optionally substituted with halogen,
(C) C.sub.1-6alkylthio, which is optionally substituted with
halogen, (D) C.sub.1-6alkylsulfonyl, (E) halogen, (F)
--NR.sub.39R.sub.40, wherein R.sub.39 and R.sub.40 are each
independently selected from: (i) hydrogen, (ii) C.sub.1-6alkyl,
(iii) C.sub.1-6alkylcarbonyl, (iv) C.sub.1-6alkylsulfonyl, (v)
arylcarbonyl, (vi) arylsulfonyl (vii) heteroarylcarbonyl, (viii)
heteroarylsulfonyl, (G) --COR.sub.41, wherein R.sub.41 is: (i)
C.sub.1-6alkyl, (ii) C.sub.1-6alkyloxy, (iii) --NR.sub.42R.sub.43,
wherein R.sub.42 and R.sub.43 are each independently selected from:
(a) hydrogen, (b) C.sub.1-6alkyl, (c) aryl, (d) heteroaryl, (iv)
hydroxyl, (H) --OR.sub.44, wherein R.sub.44 is selected from: (i)
hydrogen, (ii) C.sub.1-6alkylcarbonyl, (iii) C.sub.1-6alkylsulfonyl
(I) oxo, or wherein NR.sub.4R.sub.5 constitutes a 5-membered
heteroaryl ring containing a total of 2 nitrogen hetero atoms in
the ring; or a tautomer or pharmaceutically acceptable salt thereof
or a corresponding compound having at least one amine group
protected by an amino-protecting group.
18. A method of claim 17, wherein the compound administered is of
formula (I) wherein: X is C.sub.1-3alkyl, optionally substituted
with oxo, Y is --NH-- or --O--; R.sub.1 is: (A) aryl or heteroaryl,
each optionally and independently substituted with one or more of
the following groups: (i) C.sub.1-3alkyl, which is optionally and
independently substituted with fluorine, (ii) C.sub.1-3alkoxy,
which is optionally and independently substituted with fluorine,
(iii) C.sub.1-3alkylthio, which is optionally and independently
substituted with fluorine, (iv) arylthio, which is optionally
substituted with --NH.sub.2, (v) halogen, (vi) hydroxyl, (vii)
C.sub.1-3alkylsulfonyl, (viii) CONH.sub.2, (ix)
--NR.sub.14R.sub.15, wherein R.sub.14 and R.sub.15 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl (c)
C.sub.1-6alkylcarbonyl, (d) C.sub.1-6alkylsulfonyl, or wherein
R.sub.14 and R.sub.15 together constitute an alkylene bridge which
together with the nitrogen atom between them forms a five to
seven-membered ring, (B) cyclohexyl, which is optionally and
independently substituted with: (i) C.sub.1-3alkyl, (ii) hydroxyl;
(C) --COR.sub.23, wherein R.sub.23 is: (i) --NR.sub.24R.sub.25,
wherein R.sub.24 and R.sub.25 are each independently selected from:
(a) C.sub.1-6alkyl; or wherein R.sub.24 and R.sub.25 together
constitute a alkylene bridge which together with the nitrogen atom
between them forms a five to six-membered ring, wherein one of the
methylene groups is optionally replaced by an oxygen atom and which
ring is optionally and independently substituted with one or more
of the following groups: (i) hydroxyl, (ii) --CONH.sub.2, (D) is:
##STR00176## which is optionally substituted with halogen; (E) is
selected from the following: ##STR00177## R.sub.2 is: (A) CF.sub.3,
(B) cyano, (C) halogen, or (D) nitro, R.sub.3 is selected from the
following: ##STR00178## R.sub.4 and R.sub.5 are each independently
selected from: (A) hydrogen, (B) C.sub.1-6alkyl, which is
optionally and independently substituted with one or more of the
following groups: (i) oxo, (ii) C.sub.3-5cycloalkyl, (iii) aryl or
heteroaryl, each of which is optionally and independently
substituted with one or more of the following groups: (a)
C.sub.1-3alkyl which is optionally substituted with fluorine, (b)
--CO.sub.2H (c) halogen, (iv) NH.sub.2 (v) hydroxyl, (vi)
--CONH.sub.2, (vii) fluorine, (viii) NHCOCH.sub.3 or R.sub.4 and
R.sub.5 together constitute an alkylene bridge which together with
the nitrogen atom between them forms a four- to seven-membered
ring, wherein one of the methylene groups is optionally replaced by
an NH or oxygen atom and which ring is optionally and independently
substituted with one or more of the following groups: (A)
--CONH.sub.2, (B) NR.sub.39R.sub.40 wherein R.sub.39 and R.sub.40
are optionally and independently selected from: (i)
C.sub.1-5alkylcarbonyl (ii) C.sub.1-5alkylsulfonyl (iii)
arylcarbonyl (iv) arylsulfonyl (C) --OR.sub.44, wherein R.sub.44 is
selected from: (i) hydrogen, (ii) C.sub.1-5alkylcarbonyl (D) oxo,
or (E) fluorine or wherein NR.sub.4R.sub.5 constitute a 5-membered
heteroaryl ring containing a total of 2 nitrogen hetero atoms in
the ring, or a tautomer or pharmaceutically acceptable salt thereof
or a corresponding compound having at least one amine group
protected by an amino-protecting group.
19. A method of treating an immunological disorder in a patient
comprising administering to said patient a therapeutically
effective amount of a compound of the following formula (I):
##STR00179## wherein: X is C.sub.1-6 alkyl wherein one or two of
the methylene units can be replaced by an oxygen or sulfur atom,
and wherein the C.sub.1-6alkyl group is optionally and
independently substituted with: (A) oxo, (B) C.sub.1-6alkyl which
is optionally substituted with one or more of the following groups:
(i) hydroxyl, (ii) C.sub.1-3alkyloxy, (iii) halogen, (C)
C.sub.1-6alkyloxy, (D) C.sub.1-6alkylthio, (E) aryl (F)
--COR.sub.6, wherein R.sub.6 is: (i) C.sub.1-6alkyl, (ii)
C.sub.1-6alkyloxy, (iii) --NR.sub.7R.sub.8, wherein R.sub.7 and
R.sub.8 are each independently selected from: (a) hydrogen, (b)
C.sub.1-6alkyl, (c) aryl, (d) heteroaryl, (iv) --OH, (G) halogen,
(H)--NR.sub.9R.sub.10, wherein R.sub.9 and R.sub.10 are each
independently selected from: (i) hydrogen, (ii) C.sub.1-6alkyl,
(iii) C.sub.1-6alkylcarbonyl, (iv) C.sub.1-6alkylsulfonyl, (v)
aryl, (vi) heteroaryl; Y is --NH--, --O-- or --S--; R.sub.1 is: (A)
aryl or heteroaryl, each optionally and independently substituted
with one or more of the following groups: (i) C.sub.1-6alkyl, which
is optionally substituted one or more of the following: (a)
halogen, (b) NH.sub.2 (ii) C.sub.1-6alkoxy, which is optionally
substituted with halogen, (iii) C.sub.1-6alkylthio, which is
optionally substituted with halogen, (iv) C.sub.1-6alkylsulfonyl,
(v) cyano, (vi) halogen, (vii) hydroxyl, (viii) nitro, (ix)
--COR.sub.11, wherein R.sub.11 is: (a) C.sub.1-6alkyl, (b)
C.sub.1-6alkyloxy, (c) --OH, (d) --NR.sub.12R.sub.13, wherein
R.sub.12 and R.sub.13 are each independently selected from: (I)
hydrogen, (II) C.sub.1-6alkyl, (III) aryl, (IV) heteroaryl, (x)
--NR.sub.14R.sub.15, wherein R.sub.14 and R.sub.15 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl, (c)
C.sub.1-6alkylcarbonyl, (d) C.sub.1-6alkylsulfonyl, or wherein
R.sub.14 and R.sub.15 together constitute an alkylene bridge which
together with the nitrogen atom between them forms a five to
seven-membered ring, (xi) arylthio, arylsulfonyl or aryloxy, each
optionally and independently substituted with one or more of the
following groups: (a) C.sub.1-6alkyl, (b) C.sub.1-6alkoxy, (c)
C.sub.1-6alkylthio, (d) C.sub.1-6alkylsulfonyl, (e) cyano, (f)
halogen, (g) nitro, (h) --NR.sub.16R.sub.17, wherein R.sub.16 and
R.sub.17 are each independently selected from: (I) hydrogen, (II)
C.sub.1-6alkyl, (III) C.sub.1-6alkylcarbonyl, (IV)
C.sub.1-6alkylsulfonyl, (B) C.sub.3-6cycloalkyl which is optionally
and independently substituted with one or more of the following
groups: (i) C.sub.1-6alkyl, which is optionally substituted with
halogen, (ii) C.sub.1-6alkoxy, which is optionally substituted with
halogen, (iii) C.sub.1-6alkylthio, which is optionally substituted
with halogen, (iv) C.sub.1-6alkylsulfonyl, (v) halogen, (vi)
hydroxyl, (vii) --NR.sub.18R.sub.19, wherein R.sub.18 and R.sub.19
are each independently selected from: (a) hydrogen, (b)
C.sub.1-6alkyl, (c) C.sub.1-6alkylcarbonyl, (d)
C.sub.1-6alkylsulfonyl, (viii) --COR.sub.20, wherein R.sub.20 is:
(a) C.sub.1-6alkyl, (b) C.sub.1-6alkyloxy, (c) --OH, (d)
--NR.sub.21R.sub.22, wherein R.sub.21 and R.sub.22 are each
independently selected from: (I) hydrogen, (II) C.sub.1-6alkyl, (C)
--COR.sub.23, wherein R.sub.23 is: (i) C.sub.1-6alkyloxy, (ii)
--NR.sub.24R.sub.25, wherein R.sub.24 and R.sub.25 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl, or
wherein R.sub.24 and R.sub.25 together constitute an alkylene
bridge which together with the nitrogen atom between them forms a
four to six-membered ring, wherein one of the methylene groups is
optionally replaced by an oxygen, sulfur or NH group, and which
ring is optionally and independently substituted with one or more
of the following groups: (a) C.sub.1-6alkyloxy, (b) C.sub.1-6alkyl,
which is optionally substituted with halogen, (c) hydroxyl, (d)
halogen, (e) --COR.sub.26, wherein R.sub.26 is: (I)
C.sub.1-6alkyloxy, (II) --NR.sub.27R.sub.28, wherein R.sub.27 and
R.sub.28 are each independently selected from: a. hydrogen, b.
C.sub.1-6alkyl, c. aryl, d. heteroaryl, (D) is ##STR00180## which
is optionally substituted with halogen; (E) is selected from the
following: ##STR00181## R.sub.2 is selected from the following
groups: (A) CF.sub.3, (B) cyano, (C) halogen, (D) nitro, (E)
C.sub.1-6alkylalkynyl, (F) arylalkynyl which is optionally
substituted with one or more of the following groups: (i) halogen,
(ii) C.sub.1-6alkyl, which is optionally substituted with halogen,
R.sub.3 is selected from the following groups: ##STR00182## R.sub.4
and R.sub.5 are each independently selected from: (A) hydrogen, (B)
C.sub.1-6alkyl, optionally and independently substituted with one
or more of the following groups: (i) oxo, (ii) C.sub.3-6cycloalkyl,
(iii) C.sub.1-6alkyloxy, (iv) C.sub.1-6alkylthio, (v) --COR.sub.29,
wherein R.sub.29 is: (a) C.sub.1-6alkyl, (b) C.sub.1-6alkyloxy, (c)
--OH, (vi) --CONR.sub.30R.sub.31, wherein R.sub.30 and R.sub.31 are
each independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl,
(c) aryl, (d) heteroaryl, (vii) halogen, (viii) hydroxyl, (ix)
--NR.sub.32R.sub.33, wherein R.sub.32 and R.sub.33 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl, (c)
C.sub.1-6alkylcarbonyl, (d) C.sub.1-6alkylsulfonyl, (e) aryl, (f)
heteroaryl, (x) aryl or heteroaryl, each optionally and
independently substituted with one or more of the following groups:
(a) C.sub.1-6alkyl, which is optionally substituted with halogen,
(b) C.sub.1-6alkoxy, which is optionally substituted with halogen,
(c) C.sub.1-6alkylthio, which is optionally substituted with
halogen, (d) C.sub.1-6alkylsulfonyl, (e) cyano, (f) halogen, (g)
nitro, (h) --NR.sub.34R.sub.35, wherein R.sub.34 and R.sub.35 are
each independently selected from: (I) hydrogen, (II)
C.sub.1-6alkyl, (III) C.sub.1-6alkylcarbonyl, (IV)
C.sub.1-6alkylsulfonyl, (i) --COR.sub.36, wherein R.sub.36 is: (I)
C.sub.1-6alkyl, (II) C.sub.1-6alkyloxy, (III) --OH, (j)
--CONR.sub.37R.sub.38, wherein R.sub.37 and R.sub.38 are each
independently selected from: (I) hydrogen, (II) C.sub.1-6alkyl,
(III) aryl, (IV) heteroaryl, (C) C.sub.1-6alkylsulfonyl, (D)
arylsulfonyl, (E) aryl-C.sub.1-6alkylsulfonyl, (F)
heteroarylsulfonyl, (G) C.sub.1-6alkylcarbonyl, (H) arylcarbonyl,
(I) aryl-C.sub.1-6alkylcarbonyl, (J) heteroarylcarbonyl, (K)
C.sub.1-6alkylaminocarbonyl, or (L) heteroaryl, or wherein R.sub.4
and R.sub.5 together constitute an alkylene bridge which together
with the nitrogen atom between them forms a four- to seven-membered
ring, wherein one of the methylene groups is optionally replaced by
an oxygen, sulfur or NH group, and which ring is optionally and
independently substituted with one or more of the following groups:
(A) C.sub.1-6alkyl, which is optionally substituted with halogen,
(B) C.sub.1-6alkoxy, which is optionally substituted with halogen,
(C) C.sub.1-6alkylthio, which is optionally substituted with
halogen, (D) C.sub.1-6alkylsulfonyl, (E) halogen, (F)
--NR.sub.39R.sub.40, wherein R.sub.39 and R.sub.40 are each
independently selected from: (i) hydrogen, (ii) C.sub.1-6alkyl,
(iii) C.sub.1-6alkylcarbonyl, (iv) C.sub.1-6alkylsulfonyl, (v)
arylcarbonyl, (vi) arylsulfonyl, (vii) heteroarylcarbonyl, (viii)
heteroarylsulfonyl, (G) --COR.sub.41, wherein R.sub.1 is: (i)
C.sub.1-6alkyl, (ii) C.sub.1-6alkyloxy, (iii) --NR.sub.42R.sub.43,
wherein R.sub.42 and R.sub.43 are each independently selected from:
hydrogen, (a) C.sub.1-6alkyl, (b) aryl, (c) heteroaryl, (iv)
hydroxyl, (H) --OR.sub.44, wherein R.sub.44 is selected from: (i)
hydrogen, (ii) C.sub.1-6alkylcarbonyl, (iii) C.sub.1-6alkylsulfonyl
(I) oxo, or wherein NR.sub.4R.sub.5 constitutes a 5-membered
heteroaryl ring containing a total of 2 nitrogen hetero atoms in
the ring; or a tautomer or pharmaceutically acceptable salt thereof
or a corresponding compound having at least one amine group
protected by an amino-protecting group.
20. A method of claim 19, wherein the compound administered is of
formula (I) wherein: X is C.sub.1-3alkyl, optionally substituted
with oxo, Y is --NH-- or --O--; R.sub.1 is: (A) aryl or heteroaryl,
each optionally and independently substituted with one or more of
the following groups: (i) C.sub.1-3alkyl, which is optionally and
independently substituted with fluorine, (ii) C.sub.1-3alkoxy,
which is optionally and independently substituted with fluorine,
(iii) C.sub.1-3alkylthio, which is optionally and independently
substituted with fluorine, (iv) arylthio, which is optionally
substituted with --NH.sub.2, (v) halogen, (vi) hydroxyl, (vii)
C.sub.1-3alkylsulfonyl, (viii) CONH.sub.2, (ix)
--NR.sub.14R.sub.15, wherein R.sub.14 and R.sub.15 are each
independently selected from: (a) hydrogen, (b) C.sub.1-6alkyl (c)
C.sub.1-6alkylcarbonyl, (d) C.sub.1-6alkylsulfonyl, or wherein
R.sub.14 and R.sub.15 together constitute an alkylene bridge which
together with the nitrogen atom between them forms a five to
seven-membered ring, (B) cyclohexyl, which is optionally and
independently substituted with: (i) C.sub.1-3alkyl, (ii) hydroxyl;
(c) --COR.sub.23, wherein R.sub.23 is: (i) --NR.sub.24R.sub.25,
wherein R.sub.24 and R.sub.25 are each independently selected from:
(a) C.sub.1-6alkyl; or wherein R.sub.24 and R.sub.25 together
constitute a alkylene bridge which together with the nitrogen atom
between them forms a five to six-membered ring, wherein one of the
methylene groups is optionally replaced by an oxygen atom and which
ring is optionally and independently substituted with one or more
of the following groups: (ii) hydroxyl, (iii) --CONH.sub.2, (D) is:
##STR00183## which is optionally substituted with halogen; (E) is
selected from the following: ##STR00184## R.sub.2 is: (A) CF.sub.3,
(B) cyano, (C) halogen, or (D) nitro, R.sub.3 is selected from the
following: ##STR00185## R.sub.4 and R.sub.5 are each independently
selected from: (A) hydrogen, (B) C.sub.1-6alkyl, which is
optionally and independently substituted with one or more of the
following groups: (i) oxo, (ii) C.sub.3-5cycloalkyl, (iii) aryl or
heteroaryl, each of which is optionally and independently
substituted with one or more of the following groups: (a)
C.sub.1-3alkyl which is optionally substituted with fluorine, (b)
--CO.sub.2H (c) halogen, (iv) NH.sub.2 (v) hydroxyl, (vi)
--CONH.sub.2, (vii) fluorine, (viii) NHCOCH.sub.3 or R.sub.4 and
R.sub.5 together constitute an alkylene bridge which together with
the nitrogen atom between them forms a four- to seven-membered
ring, wherein one of the methylene groups is optionally replaced by
an NH or oxygen atom and which ring is optionally and independently
substituted with one or more of the following groups: (A)
--CONH.sub.2, (B) NR.sub.39R.sub.40 wherein R.sub.39 and R.sub.40
are optionally and independently selected from: (i)
C.sub.1-5alkylcarbonyl (ii) C.sub.1-5alkylsulfonyl (iii)
arylcarbonyl (iv) arylsulfonyl (C) --OR.sub.44, wherein R.sub.44 is
selected from: (i) hydrogen, (ii) C.sub.1-5alkylcarbonyl (D) oxo,
or (E) fluorine or wherein NR.sub.4R.sub.5 constitute a 5-membered
heteroaryl ring containing a total of 2 nitrogen hetero atoms in
the ring, or a tautomer or pharmaceutically acceptable salt thereof
or a corresponding compound having at least one amine group
protected by an amino-protecting group.
21. The method of claim 19, wherein the immunological disorder is:
an inflammatory disease, an autoimmune disease, organ and bone
marrow transplant rejection, acute or chronic inflammation, an
allergy, contact dermatitis, psoriasis, rheumatoid arthritis,
multiple sclerosis, inflammatory bowel disease, Guillain-Barre
syndrome, Crohn's disease, ulcerative colitis, graft versus host
disease or lupus erythematosus.
22. The method of claim 20, wherein the immunological disorder is:
an inflammatory disease, an autoimmune disease, organ and bone
marrow transplant rejection, acute or chronic inflammation, an
allergy, contact dermatitis, psoriasis, rheumatoid arthritis,
multiple sclerosis, inflammatory bowel disease, Guillain-Barre
syndrome, Crohn's disease, ulcerative colitis, graft versus host
disease or lupus erythematosus.
23. The method of claim 12, wherein the compound of the formula (I)
is one of the following compounds: ##STR00186## ##STR00187##
##STR00188## ##STR00189## ##STR00190## ##STR00191## ##STR00192##
##STR00193## ##STR00194## ##STR00195## ##STR00196## ##STR00197##
##STR00198## ##STR00199## ##STR00200## ##STR00201## ##STR00202##
##STR00203## ##STR00204## ##STR00205## ##STR00206## ##STR00207##
##STR00208## ##STR00209## ##STR00210## ##STR00211## ##STR00212##
##STR00213## ##STR00214## ##STR00215## ##STR00216## ##STR00217##
##STR00218## ##STR00219## ##STR00220## ##STR00221## ##STR00222##
##STR00223## ##STR00224## ##STR00225## ##STR00226## ##STR00227##
##STR00228## ##STR00229## ##STR00230## ##STR00231## ##STR00232##
##STR00233## ##STR00234## ##STR00235## ##STR00236## ##STR00237##
##STR00238## ##STR00239## ##STR00240## ##STR00241## ##STR00242##
##STR00243## ##STR00244## ##STR00245## ##STR00246## ##STR00247##
##STR00248## ##STR00249## ##STR00250## ##STR00251## ##STR00252##
##STR00253## ##STR00254## ##STR00255## ##STR00256## ##STR00257##
##STR00258## ##STR00259## ##STR00260## ##STR00261## ##STR00262##
##STR00263## ##STR00264##
24. The method of claim 12, wherein the compound of the formula (I)
is one of the following compounds:
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-fluoro-3-(trifluorom-
ethyl)benzyl]-5-nitropyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-chloro-6-fluoro-3-me-
thylbenzyl)-5-nitropyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(1-naphthylmethyl)-5-ni-
tropyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-{2-[(trifluorom-
ethyl)thio]benzyl}pyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-chloro-N.sup.2-[2-(trifluorom-
ethoxy)benzyl]pyrimidine-2,4-diamine;
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)azetidin-3-one;
N.sup.4-{[trans-4-(3-aminoazetidin-1-yl)cyclohexyl]methyl}-5-nitro-N.sup.-
2-[2-(trifluoromethoxy)benzyl]-pyrimidine-2,4-diamine;
N-[1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-
-yl)amino]methyl}-cyclohexyl)azetidin-3-yl]methanesulfonamide;
N-[1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-
-yl)amino]methyl}-cyclohexyl)azetidin-3-yl]acetamide;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]pyrimidine-2,4-diamine;
4-{[(trans-4-aminocyclohexyl)methyl]amino}-2-{[2-(trifluoromethoxy)benzyl-
]amino}pyrimidine-5-carbonitrile;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-{2-[(2-aminophenyl)thio-
]benzyl}-5-nitropyrimidine-2,4-diamine;
5-nitro-N.sup.4-[(trans-4-pyrrolidin-1-ylcyclohexyl)methyl]-N.sup.2-[2-(t-
rifluoromethoxy)benzyl]pyrimidine-2,4-diamine;
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)pyrrolidin-3-ol;
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-5-nitro-N.sup.2-[2-(tri-
fluoromethoxy)benzyl]pyrimidine-2,4-diamine;
N.sup.4-{[trans-4-(dimethylamino)cyclohexyl]methyl}-5-nitro-N.sup.2-[2-(t-
rifluoromethoxy)benzyl]pyrimidine-2,4-diamine;
4-({[trans-4-(dimethylamino)cyclohexyl]methyl}amino)-2-{[2-(trifluorometh-
oxy)benzyl]-amino}pyrimidine-5-carbonitrile;
5-nitro-N.sup.4-({trans-4-[(pyridin-3-ylmethyl)amino]cyclohexyl}methyl)-N-
.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine;
5-nitro-N.sup.4-({trans-4-[(pyridin-4-ylmethyl)amino]cyclohexyl}methyl)-N-
.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine;
2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-y-
l)amino]methyl}-cyclohexyl)amino]ethanol;
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)azetidin-3-ol;
5-nitro-N.sup.4-({trans-4-[(pyridin-2-ylmethyl)amino]cyclohexyl}methyl)-N-
.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine;
N.sup.2-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}-cyclohexyl)glycinamide;
N.sup.4-[(trans-4-{[(1,5-dimethyl-1H-pyrazol-4-yl)methyl]amino}cyclohexyl-
)methyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamin-
e;
N.sup.4-[(trans-4-{[(1-methyl-1H-pyrazol-4-yl)methyl]amino}cyclohexyl)m-
ethyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(2-isopropoxypyridin-3-
-yl)methyl]-5-nitropyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-{[2-(2,2,2-trif-
luoroethoxy)pyridin-3-yl]methyl}pyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(5-fluoro-2-methoxypyr-
idin-3-yl)methyl]-5-nitropyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(difluoromethoxy)ben-
zyl]-5-nitropyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-chlorobenzyl)-5-nitr-
opyrimidine-2,4-diamine;
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-5-nitro-N.sup.2-{[2-(2,-
2,2-trifluoroethoxy)pyridin-3-yl]methyl}pyrimidine-2,4-diamine;
N.sup.4-{[trans-4-(3-fluoroazetidin-1-yl)cyclohexyl]methyl}-5-nitro-N.sup-
.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine;
N.sup.4-{[trans-4-(ethylamino)cyclohexyl]methyl}-5-nitro-N.sup.2-[2-(trif-
luoromethoxy)benzyl]pyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-chloro-3-(dimethylam-
ino)benzyl]-5-nitropyrimidine-2,4-diamine;
1-(trans-4-{[(2-{[2-chloro-3-(dimethylamino)benzyl]amino}-5-nitropyrimidi-
n-4-yl)amino]methyl}cyclohexyl)azetidin-3-ol;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(5-chloro-2-methoxybenz-
yl)-5-nitropyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-methoxybenzyl)-5-nit-
ropyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[5-chloro-2-(trifluorom-
ethoxy)benzyl]-5-nitropyrimidine-2,4-diamine;
N.sup.2-[5-chloro-2-(trifluoromethoxy)benzyl]-5-nitro-N.sup.4-[(trans-4-p-
yrrolidin-1-ylcyclohexyl)-methyl]pyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(2-methoxypyridin-3-yl-
)methyl]-5-nitropyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-{[4-(trifluorom-
ethyl)pyridin-3-yl]methyl}pyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[5-chloro-2-(methylsulf-
onyl)benzyl]-5-nitropyrimidine-2,4-diamine;
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-N.sup.2-[(4-chloropyrid-
in-3-yl)methyl]-5-nitropyrimidine-2,4-diamine;
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-5-nitro-N.sup.2-{[4-(tr-
ifluoromethyl)pyridin-3-yl]methyl}pyrimidine-2,4-diamine;
1-[trans-4-({[5-nitro-2-({[4-(trifluoromethyl)pyridin-3-yl]methyl}amino)p-
yrimidin-4-yl]amino}methyl)cyclohexyl]azetidin-3-ol;
(2R)-1-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]propan-2-ol;
(2S)-1-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]propan-2-ol;
3-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-y-
l)amino]methyl}cyclohexyl)amino]propan-1-ol;
(2R)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]propan-1-ol; or
(2S)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]propan-1-ol.
Description
[0001] Benefit of U.S. Provisional Application Ser. No. 60/586,289,
filed on Jul. 8, 2004 is hereby claimed.
FIELD OF THE INVENTION
[0002] This invention relates to substituted pyrimidine derivatives
which are useful as inhibitors of PKC-theta and are thus useful for
treating a variety of diseases and disorders that are mediated or
sustained through the activity of PKC-theta, including
immunological disorders and type II diabetes. This invention also
relates to pharmaceutical compositions comprising these compounds,
methods of using these compounds in the treatment of various
diseases and disorders, processes for preparing these compounds and
intermediates useful in these processes.
BACKGROUND OF THE INVENTION
[0003] The protein kinase C family is a group of serine/threonine
kinases that is comprised of twelve related isoenzymes. These
kinases are expressed in a wide range of tissues and cell types.
Its members are encoded by different genes and are sub-classified
according to their requirements for activation. The classical PKC
enzymes (cPKC) require diacylglycerol (DAG), phosphatidylserine
(PS) and calcium for activation. The novel PKC's (nPKC) require DAG
and PS but are calcium independent. The atypical PKC's (aPKC) do
not require calcium or DAG.
[0004] PKC-theta is a member of the nPKC sub-family. It has a
restricted expression pattern, found predominantly in T cells and
skeletal muscle. Upon T cell activation, a supramolecular
activation complex (SMAC) forms at the site of contact between the
T cell and antigen presenting cell (APC). PKC-theta is the only PKC
isoform found to localize at the SMAC (C. Monks et al., Nature,
1997, 385, 83), placing it in proximity with other signaling
enzymes that mediate T cell activation processes. In another study
(G. Baier-Bitterlich et al., Mol. Cell. Biol., 1996, 16, 842) the
role of PKC-theta in the activation of AP-1, a transcription factor
important in the activation of the IL-2 gene, was confirmed. In
unstimulated T cells, constitutively active PKC-theta stimulated
AP-1 activity while in cells with dominant negative PKC-theta, AP-1
activity was not induced upon activation by PMA. Other studies
showed that PKC-theta, via activation of I.kappa.B kinase beta,
mediates activation of NF-.kappa.B induced by T cell receptor/CD28
co-stimulation (N. Coudronniere et al., Proc. Nat. Acad. Sci.
U.S.A., 2000, 97, 3394; X. Lin et al., Moll. Cell. Biol., 2000, 20,
2933). Proliferation of peripheral T cells from PKC-theta knockout
mice, in response to T cell receptor (TCR)/CD28 stimulation was
greatly diminished compared to T cells from wild type mice. In
addition, the amount of IL-2 released from the T cells was also
greatly reduced (Z. Sun et al., Nature, 2000, 404, 402). Otherwise,
the PKC-theta knockout mice seemed normal and were fertile.
[0005] The studies cited above and other studies confirm the
critical role of PKC-theta in T cell activation and subsequent
release of cytokines such as IL-2 and T cell proliferation (A.
Altman et al., Immunology Today, 2000, 21, 567). Thus an inhibitor
of PKC-theta would be of therapeutic benefit in treating
immunological disorders and other diseases mediated by the
inappropriate activation of T cells.
[0006] It has been well established that T cells play an important
role in regulating the immune response (Powrie and Coffman,
Immunology Today, 1993, 14, 270). Indeed, activation of T cells is
often the initiating event in immunological disorders. Following
activation of the TCR, there is an influx of calcium that is
required for T cell activation. Upon activation, T cells produce
cytokines, including IL-2, leading to T cell proliferation,
differentiation, and effector function. Clinical studies with
inhibitors of IL-2 have shown that interference with T cell
activation and proliferation effectively suppresses immune response
in vivo (Waldmann, Immunology Today, 1993, 14, 264).
[0007] Accordingly, agents that inhibit T lymphocyte activation and
subsequent cytokine production are therapeutically useful for
selectively suppressing the immune response in a patient in need of
such immunosuppression and therefore are useful in treating
immunological disorders such as autoimmune and inflammatory
diseases.
[0008] In addition, PKC theta activation has been shown to be
associated with insulin resistance in skeletal muscle (M. E.
Griffen et al., Diabetes, 1999, 48, 1270). Therefore inhibitors of
PKC-theta may also be useful for treating type B diabetes.
[0009] Dahmann et al, U.S. application Ser. No. 10/271,763, filed
Oct. 16, 2002, (now U.S. Patent Application Publication No.
2003/0171359 A1) discloses pyrimidine derivatives as inhibitors of
various protein kinases such as SRC kinase, PLK kinase and
particularly cyclin-dependent kinases (CDKs) and Aurora B. WO
00/75113 and U.S. Pat. No. 6,432,963 describe pyrimidine
carboxamides as inhibitors of Syk tyrosine kinase. WO 01/00213
discloses heteroaryl substituted pyrimidines as SRC kinase
inhibitors. WO 97/19065 describes substituted 2-anilinopyrimidine
compounds as inhibitors of certain protein kinases. WO 02/096887
and WO 02/096888 both disclose 2-anilinopyrimidine derivatives as
inhibitors of cyclin-dependent kinases. WO 03/106451 discloses
certain substituted diaminopyrimidine compounds as inhibitors of
PKC-theta.
[0010] There is a continuing need in the art for compounds that are
potent and selective inhibitors of PKC-theta.
BRIEF SUMMARY OF THE INVENTION
[0011] In a general aspect, the present invention is directed to
the compounds of the following formula (I):
##STR00002##
wherein R.sub.1, R.sub.2 and R.sub.3 are as defined herein, as well
as the tautomers, pharmaceutically acceptable salts, solvates, and
amino-protected derivatives thereof. It has been found that the
compounds of formula (I) have valuable pharmacological properties,
particularly an inhibiting activity on PKC-theta. Many of the
compounds of the invention are not only potent inhibitors of
PKC-theta but are also selective for the inhibition of PKC-theta as
compared to one or more other protein kinases.
[0012] In another aspect, the present invention is directed to a
method of inhibiting PKC-theta activity in a patient comprising
administering to the patient a compound of the present invention as
described above.
[0013] In another aspect, the present invention is directed to a
method of treating a disease or disorder associated with the
activation of T cells comprising administering to a patient in need
of such treatment a compound of the present invention as described
above.
[0014] In another aspect, the present invention is directed to a
method of treating an immunological disorder comprising
administering to a patient in need of such treatment a compound of
the present invention as described above. Examples of such
immunological disorders that may be treated include, for example,
inflammatory diseases, autoimmune diseases, organ and bone marrow
transplant rejection and other disorders associated with T cell
mediated immune response, including acute or chronic inflammation,
allergies, contact dermatitis, psoriasis, rheumatoid arthritis,
multiple sclerosis, type I diabetes, inflammatory bowel disease,
Guillain-Barre syndrome, Crohn's disease, ulcerative colitis, graft
versus host disease (and other forms of organ or bone marrow
transplant rejection) and lupus erythematosus.
[0015] In another aspect, the present invention is directed to a
method of treating type II diabetes comprising administering to a
patient in need of such treatment a compound of the present
invention as described above.
[0016] In yet additional aspects, the present invention is directed
to pharmaceutical compositions comprising the above-mentioned
compounds, processes for preparing the above-mentioned compounds
and intermediates used in these processes.
DETAILED DESCRIPTION OF THE INVENTION
Definition of Terms and Conventions Used
[0017] Terms not specifically defined herein should be given the
meanings that would be given to them by one of skill in the art in
light of the disclosure and the context. As used in the present
specification and claims, however, unless specified to the
contrary, the following terms have the meaning indicated and the
following conventions are adhered to.
A. Chemical Nomenclature, Terms, and Conventions
[0018] In general, for groups comprising two or more subgroups, the
last named group is the radical attachment point, for example,
"alkylaryl" means a monovalent radical of the formula Alk-Ar-,
while "arylalkyl" means a monovalent radical of the formula Ar-Alk-
(where Alk is an alkyl group and Ar is an aryl group). Furthermore,
the use of a term designating a monovalent radical where a divalent
radical is appropriate shall be construed to designate the
respective divalent radical and vice versa. Unless otherwise
specified, conventional definitions of terms control and
conventional stable atom valences are presumed and achieved in all
formulas and groups.
[0019] All references to a chemical group being "substituted with"
another chemical group shall be understood to mean the first
chemical group can be substituted with one or more of the second
chemical group, with the exception of any substitution pattern that
is not physically or chemically possible or results in a unstable
structure or compound. For example, the phrase "C.sub.1-6 alkyl,
which is optionally substituted with halogen" shall mean a
C.sub.1-6 alkyl group having one or multiple halogen substituents
being the same or different from each other.
[0020] All alkyl groups shall be understood as being branched or
unbranched unless otherwise specified. Other more specific
definitions are as follows:
[0021] The term "halogen" as used in the present specification
shall be understood to mean bromine, chlorine, fluorine or
iodine.
[0022] The term "heteroaryl" refers to a stable 5 or 6 membered,
monocyclic aromatic heterocycle radical, wherein the heterocycle
radical is optionally fused to either an aryl, e.g. benzene, or to
a second 5 or 6 membered, monocyclic aromatic heterocycle to form
in each case a bicyclic heteroaryl group. Each heterocycle consists
of carbon atoms and from 1 to 3 heteroatoms chosen from nitrogen,
oxygen and sulfur. The heterocycle may be attached by any atom of
the cycle, which results in the creation of a stable structure.
Example "heteroaryl" radicals include, pyridyl, pyrimidinyl,
pyrazinyl, pyridazinyl, pyrrolyl, imidazolyl, pyrazolyl, thienyl,
furyl, isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, oxadiazolyl,
thiadiazolyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl,
benzofuranyl, benzoxazolyl, benzisoxazolyl, benzopyrazolyl,
benzothiofuranyl, benzothiazolyl, quinazolinyl and indazolyl.
[0023] The term "aryl" shall be understood to mean a 6-10 membered
monocyclic or bicyclic aromatic carbocycle, and includes, for
example, phenyl and naphthyl; other terms comprising "aryl" will
have the same definition for the aryl component, and examples of
these moieties include: arylalkyl, aryloxy or arylthio.
[0024] The term "oxo" refers to a double-bonded oxygen group
(.dbd.O).
[0025] The phrases "wherein each of the C.sub.1-6alkyl groups",
"wherein each of the C.sub.1-8alkyl groups" or "wherein each of the
aryl groups" or similar language in a definition is intended to
refer to the indicated groups when either alone or as part of
another chemical group if such combined groups are provided for in
a definition. For example, the language "wherein each of the
C.sub.1-6alkyl groups" refers to C.sub.1-6alkyl groups as well as
C.sub.1-6alkyl groups when attached to other groups, e.g., the
C.sub.1-6alkyl portion of a C.sub.1-6alkyloxy or
aryl-C.sub.1-6alkyl group, if such groups are provided for in a
definition.
[0026] The term "amino protected derivatives" shall be understood
to mean compounds of formula (I) wherein one or more of the amine
groups are protected by suitable amino protecting groups. Amino
protecting groups that may be used include, for example,
alkoxycarbonyl groups, such as tert-butyloxycarbonyl (Boc) and
ethoxycarbonyl, Mannich bases, Schiff bases and amino acids. As
would be understood by a person skilled in the art, such amino
protected compounds may be useful as intermediates in the
preparation of other compounds of formula (I), e.g., as described
in the synthetic processes below, and/or may themselves be useful
as prodrugs that can be administered to a patient to be converted
in vivo into a PKC-theta inhibitor having the resulting
pharmacologic and therapeutic effects expected from the inhibition
of PKC-theta in a patient.
[0027] The term "pharmaceutically acceptable salts" include those
derived from pharmaceutically acceptable inorganic and organic
acids and bases. Examples of suitable acids include hydrochloric,
hydrobromic, carbonic, sulfuric, nitric, perchloric, fumaric,
maleic, phosphoric, glycolic, lactic, salicylic, succinic,
toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic,
formic, benzoic, malonic, naphthalene-2-sulfonic and
benzenesulfonic acids. Other acids, such as oxalic acid, while not
themselves pharmaceutically acceptable, may be employed in the
preparation of salts useful as intermediates in obtaining the
compounds of this invention and their pharmaceutically acceptable
acid addition salts. Salts derived from appropriate bases include
alkali metal (e.g., sodium), alkaline earth metal (e.g.,
magnesium), ammonium and N--(C.sub.1-4 alkyl).sub.4.sup.+
salts.
[0028] The term "solvate" means a physical association of a
compound with one or more solvent molecules or a complex of
variable stoichiometry formed by a solute (for example, a compound
of Formula (I)) and a solvent, for example, water, EtOH, or acetic
acid. This physical association may involve varying degrees of
ionic and covalent bonding, including hydrogen bonding. In certain
instances, the solvate will be capable of isolation, for example,
when one or more solvent molecules are incorporated in the crystal
lattice of the crystalline solid. In general, the solvents selected
do not interfere with the biological activity of the solute.
Solvates encompasses both solution-phase and isolatable solvates.
Representative solvates include hydrates, EtOHates, MeOHates, and
the like.
[0029] The term "hydrate" means a solvate wherein the solvent
molecule(s) is/are H.sub.2O.
[0030] The term "compounds of the invention" and equivalent
expressions are meant to embrace compounds of Formula (I) as herein
described, including the tautomers, pharmaceutically acceptable
salts, solvates, and amino-protected derivatives thereof, where the
context so permits. In general, the compounds of the invention and
the formulas designating the compounds of the invention are
understood to only include the stable compounds thereof and exclude
unstable compounds, even if an unstable compound might be
considered to be literally embraced by the compound formula.
[0031] The term "stable compound" means a compound that is
sufficiently robust to survive isolation to a useful degree of
purity from a reaction mixture, and formulation into an efficacious
therapeutic agent. For example, a compound which would have a
"dangling valency" is not a compound contemplated by the
invention.
[0032] Specific compounds of the present invention may be
identified in the present specification by chemical name and/or
chemical structure. In the event of any conflict between the
chemical name and chemical structure, the chemical structure will
control.
B. Isomer Terms and Conventions
[0033] In general, all tautomeric and isomeric forms and mixtures
thereof, for example, individual geometric isomers, stereoisomers,
enantiomers, diastereomers, racemates, racemic or non-racemic
mixtures of stereoisomers, mixtures of diastereomers, or mixtures
of any of the foregoing forms of a chemical structure or compound
is intended, unless the specific stereochemistry or isomeric form
is specifically indicated in the compound name or structure.
[0034] It is well-known in the art that the biological and
pharmacological activity of a compound is sensitive to the
stereochemistry of the compound. Thus, for example, enantiomers
often exhibit strikingly different biological activity including
differences in pharmacokinetic properties, including metabolism,
protein binding, and the like, and pharmacological properties,
including the type of activity displayed, the degree of activity,
toxicity, and the like. Thus, one skilled in the art will
appreciate that one enantiomer may be more active or may exhibit
beneficial effects when enriched relative to the other enantiomer
or when separated from the other enantiomer. Additionally, one
skilled in the art would know how to separate, enrich, or
selectively prepare the enantiomers of the compounds of the present
invention from this disclosure and the knowledge in the art.
[0035] Preparation of pure stereoisomers, e.g. enantiomers and
diastereomers, or mixtures of desired enantiomeric excess (ee) or
enantiomeric purity, are accomplished by one or more of the many
methods of (a) separation or resolution of enantiomers, or (b)
enantioselective synthesis known to those of skill in the art, or a
combination thereof. These resolution methods generally rely on
chiral recognition and include, for example, chromatography using
chiral stationary phases, enantioselective host-guest complexation,
resolution or synthesis using chiral auxiliaries, enantioselective
synthesis, enzymatic and nonenzymatic kinetic resolution, or
spontaneous enantioselective crystallization. Such methods are
disclosed generally in Chiral Separation Techniques: A Practical
Approach (2nd Ed.), G. Subramanian (ed.), Wiley-VCH, 2000; T. E.
Beesley and R. P. W. Scott, Chiral Chromatography, John Wiley &
Sons, 1999; and Satinder Ahuja, Chiral Separations by
Chromatography, Am. Chem. Soc., 2000. Furthermore, there are
equally well-known methods for the quantitation of enantiomeric
excess or purity, for example, GC, HPLC, CE, or NMR, and assignment
of absolute configuration and conformation, for example, CD ORD,
X-ray crystallography, or NMR
C. Pharmaceutical Administration Terms and Conventions
[0036] The term "patient" includes both human and non-human
mammals.
[0037] The term "therapeutically effective amount" means an amount
of a compound according to the invention which, when administered
to a patient in need thereof, is sufficient to effect treatment for
disease-states, conditions, or disorders for which the compounds
have utility. Such an amount would be sufficient to elicit the
biological or medical response of a tissue, system, or patient that
is sought by a researcher or clinician. The amount of a compound of
according to the invention which constitutes a therapeutically
effective amount will vary depending on such factors as the
compound and its biological activity, the composition used for
administration, the time of administration, the route of
administration, the rate of excretion of the compound, the duration
of treatment, the type of disease-state or disorder being treated
and its severity, drugs used in combination with or coincidentally
with the compounds of the invention, and the age, body weight,
general health, sex, and diet of the patient. Such a
therapeutically effective amount can be determined routinely by one
of ordinary skill in the art having regard to their own knowledge,
the state of the art, and this disclosure.
[0038] The phrase "disease or disorder associated with the
activation of T cells" and similar expressions mean that the
activation of T cells is a contributing factor to either the origin
or continuation of the disease or disorder in the patient.
EMBODIMENTS OF THE INVENTION
[0039] In its broadest generic aspect, the invention provides novel
compounds of the formula (I) below
##STR00003##
wherein: [0040] X is C.sub.1-6alkyl wherein one or two of the
methylene units can be replaced by an oxygen or sulfur atom, and
wherein the C.sub.1-6alkyl group is optionally and independently
substituted with: [0041] (A) oxo, [0042] (B) C.sub.1-6alkyl which
is optionally substituted with one or more of the following groups:
[0043] (i) hydroxyl, [0044] (ii) C.sub.1-3alkyloxy, [0045] (iii)
halogen, [0046] (C) C.sub.1-6alkyloxy, [0047] (D)
C.sub.1-6alkylthio, [0048] (E) aryl [0049] (F) --COR.sub.6, wherein
R.sub.6 is: [0050] (i) C.sub.1-6alkyl, [0051] (ii)
C.sub.1-6alkyloxy, [0052] (iii) --NR.sub.7R.sub.8, wherein R.sub.7
and R.sub.8 are each independently selected from: [0053] (a)
hydrogen, [0054] (b) C.sub.1-6alkyl, [0055] (c) aryl, [0056] (d)
heteroaryl, [0057] (iv) --OH, [0058] (G) halogen, [0059] (H)
--NR.sub.9R.sub.10, wherein R.sub.9 and R.sub.10 are each
independently selected from: [0060] (i) hydrogen, [0061] (ii)
C.sub.1-6allyl, [0062] (iii) C.sub.1-6alkylcarbonyl, [0063] (iv)
C.sub.1-6alkylsulfonyl, [0064] (v) aryl, [0065] (vi) heteroaryl;
[0066] Y is --NH--, --O--, or --S--; [0067] R.sub.1 is: [0068] (A)
aryl or heteroaryl, each optionally and independently substituted
with one or more of the following groups: [0069] (i)
C.sub.1-6alkyl, which is optionally substituted one or more of the
following: [0070] (a) halogen, [0071] (b) NH.sub.2 [0072] (ii)
C.sub.1-6alkoxy, which is optionally substituted with halogen,
[0073] (iii) C.sub.1-6alkylthio, which is optionally substituted
with halogen, [0074] (iv) C.sub.1-6alkylsulfonyl, [0075] (v) cyano,
[0076] (vi) halogen, [0077] (vii) hydroxyl, [0078] (viii) nitro,
[0079] (ix) --COR.sub.11, wherein R.sub.11 is: [0080] (a)
C.sub.1-6alkyl, [0081] (b) C.sub.1-6alkyloxy, [0082] (c) --OH,
[0083] (d) --NR.sub.12R.sub.13, wherein R.sub.12 and R.sub.13 are
each independently selected from: [0084] (I) hydrogen, [0085] (II)
C.sub.1-6alkyl, [0086] (III) aryl, [0087] (IV) heteroaryl, [0088]
(x) --NR.sub.14R.sub.15, wherein R.sub.14 and R.sub.15 are each
independently selected from: [0089] (a) hydrogen, [0090] (b)
C.sub.1-6alkyl, [0091] (c) C.sub.1-6alkylcarbonyl, [0092] (d)
C.sub.1-6alkylsulfonyl, [0093] or wherein R.sub.14 and R.sub.15
together constitute an alkylene bridge which together with the
nitrogen atom between them forms a five to seven-membered ring,
[0094] (xi) arylthio, arylsulfonyl or aryloxy, each optionally and
independently substituted with one or more of the following groups:
[0095] (a) C.sub.1-6alkyl, [0096] (b) C.sub.1-6alkoxy, [0097] (c)
C.sub.1-6alkylthio, [0098] (d) C.sub.1-6alkylsulfonyl, [0099] (e)
cyano, [0100] (f) halogen, [0101] (g) nitro, [0102] (h)
--NR.sub.16R.sub.17, wherein R.sub.16 and R.sub.17 are each
independently selected from: [0103] (I) hydrogen, [0104] (II)
C.sub.1-6alkyl, [0105] (III) C.sub.1-6alkylcarbonyl, [0106] (IV)
C.sub.1-6alkylsulfonyl, [0107] (B) C.sub.3-6cycloalkyl which is
optionally and independently substituted with one or more of the
following groups: [0108] (i) C.sub.1-6alkyl, which is optionally
substituted with halogen, [0109] (ii) C.sub.1-6alkoxy, which is
optionally substituted with halogen, [0110] (iii)
C.sub.1-6alkylthio, which is optionally substituted with halogen,
[0111] (iv) C.sub.1-6alkylsulfonyl, [0112] (v) halogen, [0113] (vi)
hydroxyl, [0114] (vii) --NR.sub.18R.sub.19, wherein R.sub.18 and
R.sub.19 are each independently selected from: [0115] (a) hydrogen,
[0116] (b) C.sub.1-6alkyl, [0117] (c) C.sub.1-6alkylcarbonyl,
[0118] (d) C.sub.1-6alkylsulfonyl, [0119] (viii) --COR.sub.20,
wherein R.sub.20 is: [0120] (a) C.sub.1-6allyl, [0121] (b)
C.sub.1-6alkyloxy, [0122] (c) --OH, [0123] (d) --NR.sub.21R.sub.22,
wherein R.sub.21 and R.sub.22 are each independently selected from:
[0124] (I) hydrogen, [0125] (II) C.sub.1-6alkyl, [0126] (C)
--COR.sub.23, wherein R.sub.23 is: [0127] (i) C.sub.1-6alkyloxy,
[0128] (ii) --NR.sub.24R.sub.25, wherein R.sub.24 and R.sub.25 are
each independently selected from: [0129] (a) hydrogen, [0130] (b)
C.sub.1-6alkyl, [0131] or wherein R.sub.24 and R.sub.25 together
constitute an alkylene bridge which together with the nitrogen atom
between them forms a four to six-membered ring, wherein one of the
methylene groups is optionally replaced by an oxygen, sulfur or NH
group, and which ring is optionally and independently substituted
with one or more of the following groups: [0132] (a)
C.sub.1-6alkyloxy, [0133] (b) C.sub.1-6alkyl, which is optionally
substituted with halogen, [0134] (c) hydroxyl, [0135] (d) halogen,
[0136] (e) --COR.sub.26, wherein R.sub.26 is: [0137] (I)
C.sub.1-6alkyloxy, [0138] (II) --NR.sub.27R.sub.28, wherein
R.sub.27 and R.sub.28 are each independently selected from: a.
hydrogen, b. C.sub.1-6alkyl, c. aryl, d. heteroaryl, [0139] (D)
is:
[0139] ##STR00004## [0140] which is optionally substituted with
halogen; [0141] (E) is selected from the following:
[0141] ##STR00005## [0142] R.sub.2 is selected from the following
groups: [0143] (A) CF.sub.3, [0144] (B) cyano, [0145] (C) halogen,
[0146] (D) nitro, [0147] (E) C.sub.1-6alkylalkynyl, [0148] (F)
arylalkynyl which is optionally substituted with one or more of the
following groups: [0149] (i) halogen, [0150] (ii) C.sub.1-6alkyl,
which is optionally substituted with halogen, [0151] R.sub.3 is
selected from the following groups:
[0151] ##STR00006## [0152] R.sub.4 and R.sub.5 are each
independently selected from: [0153] (A) hydrogen, [0154] (B)
C.sub.1-6alkyl, optionally and independently substituted with one
or more of the following groups: [0155] (i) oxo, [0156] (ii)
C.sub.3-6cycloalkyl, [0157] (iii) C.sub.1-6allyloxy, [0158] (iv)
C.sub.1-6alkylthio, [0159] (v) --COR.sub.29 wherein R.sub.29 is:
[0160] (a) C.sub.1-6alkyl, [0161] (b) C.sub.1-6alkyloxy, [0162] (c)
--OH, [0163] (vi) --CONR.sub.30R.sub.31, wherein R.sub.30 and
R.sub.31 are each independently selected from: [0164] (a) hydrogen,
[0165] (b) C.sub.1-6alkyl, [0166] (c) aryl, [0167] (d) heteroaryl,
[0168] (vii) halogen, [0169] (viii) hydroxyl, [0170] (ix)
--NR.sub.32R.sub.33, wherein R.sub.32 and R.sub.33 are each
independently selected from: [0171] (a) hydrogen, [0172] (b)
C.sub.1-6alkyl, [0173] (c) C.sub.1-6alkylcarbonyl, [0174] (d)
C.sub.1-6alkylsulfonyl, [0175] (e) aryl, [0176] (f) heteroaryl,
[0177] (x) aryl or heteroaryl, each optionally and independently
substituted with one or more of the following groups: [0178] (a)
C.sub.1-6alkyl, which is optionally substituted with halogen,
[0179] (b) C.sub.1-6alkoxy, which is optionally substituted with
halogen, [0180] (c) C.sub.1-6alkylthio, which is optionally
substituted with halogen, [0181] (d) C.sub.1-6alkylsulfonyl, [0182]
(e) cyano, [0183] (f) halogen, [0184] (g) nitro, [0185] (h)
--NR.sub.34R.sub.35, wherein R.sub.34 and R.sub.35 are each
independently selected from: [0186] (I) hydrogen, [0187] (II)
C.sub.1-6alkyl, [0188] (III) C.sub.1-6alkylcarbonyl, [0189] (IV)
C.sub.1-6alkylsulfonyl, [0190] (i) --COR.sub.36, wherein R.sub.36
is: [0191] (I) C.sub.1-6alkyl, [0192] (II) C.sub.1-6alkyloxy,
[0193] (III) --OH, [0194] (j) CONR.sub.37R.sub.38, wherein R.sub.37
and R.sub.38 are each independently selected from: [0195] (I)
hydrogen, [0196] (II) C.sub.1-6allyl, [0197] (III) aryl, [0198]
(IV) heteroaryl, [0199] (C) C.sub.1-6alkylsulfonyl, [0200] (D)
arylsulfonyl, [0201] (E) aryl-C.sub.1-6alkylsulfonyl, [0202] (F)
heteroarylsulfonyl, [0203] (G) C.sub.1-6alkylcarbonyl, [0204] (H)
arylcarbonyl, [0205] (I) aryl-C.sub.1-6alkylcarbonyl, [0206] (J)
heteroarylcarbonyl, [0207] (K) C.sub.1-6alkylaminocarbonyl, or
[0208] (L) heteroaryl, [0209] or wherein R.sub.4 and R.sub.5
together constitute an alkylene bridge which together with the
nitrogen atom between them forms a four- to seven-membered ring,
wherein one of the methylene groups is optionally replaced by an
oxygen, sulfur or NH group, and which ring is optionally and
independently substituted with one or more of the following groups:
[0210] (A) C.sub.1-6alkyl, which is optionally substituted with
halogen, [0211] (B) C.sub.1-6alkoxy, which is optionally
substituted with halogen, [0212] (C) C.sub.1-6alkylthio, which is
optionally substituted with halogen, [0213] (D)
C.sub.1-6alkylsulfonyl, [0214] (E) halogen, [0215] (F)
--NR.sub.39R.sub.40, wherein R.sub.39 and R.sub.40 are each
independently selected from: [0216] (i) hydrogen, [0217] (ii)
C.sub.1-6alkyl, [0218] (iii) C.sub.1-6alkylcarbonyl, [0219] (iv)
C.sub.1-6alkylsulfonyl, [0220] (v) arylcarbonyl, [0221] (vi)
arylsulfonyl [0222] (vii) heteroarylcarbonyl, [0223] (viii)
heteroarylsulfonyl, [0224] (G) --COR.sub.41, wherein R.sub.41 is:
[0225] (i) C.sub.1-6alkyl, [0226] (ii) C.sub.1-6alkyloxy, [0227]
(iii) --NR.sub.42R.sub.43, wherein R.sub.42 and R.sub.43 are each
independently selected from: [0228] (a) hydrogen, [0229] (b)
C.sub.1-6alkyl, [0230] (c) aryl, [0231] (d) heteroaryl, [0232] (iv)
hydroxyl, [0233] (H) --OR.sub.44, wherein R.sub.44 is selected
from: [0234] (i) hydrogen, [0235] (ii) C.sub.1-6alkylcarbonyl,
[0236] (iii) C.sub.1-6alkylsulfonyl [0237] (I) oxo, [0238] or
wherein NR.sub.4R.sub.5 constitutes a 5-membered heteroaryl ring
containing a total of 2 nitrogen hetero atoms in the ring; or a
tautomer, pharmaceutically acceptable salt, solvate or
amino-protected derivative thereof.
[0239] In another embodiment there are provided compounds of
formula (I) as described above and wherein [0240] X is
C.sub.1-3alkyl, optionally substituted with oxo, [0241] Y is --NH--
or --O--; [0242] R.sub.1 is: [0243] (A) aryl or heteroaryl, each
optionally and independently substituted with one or more of the
following groups: [0244] (i) C.sub.1-3alkyl, which is optionally
and independently substituted with fluorine, [0245] (ii)
C.sub.1-3alkoxy, which is optionally and independently substituted
with fluorine, [0246] (iii) C.sub.1-3alkylthio, which is optionally
and independently substituted with fluorine, [0247] (iv) arylthio,
which is optionally substituted with --NH.sub.2, [0248] (v)
halogen, [0249] (vi) hydroxyl, [0250] (vii) C.sub.1-3alkylsulfonyl,
[0251] (viii) CONH.sub.2, [0252] (ix) --NR.sub.14R.sub.15, wherein
R.sub.14 and R.sub.15 are each independently selected from: [0253]
(a) hydrogen, [0254] (b) C.sub.1-6alkyl [0255] (c)
C.sub.1-6alkylcarbonyl, [0256] (d) C.sub.1-6alkylsulfonyl, [0257]
or wherein R.sub.14 and R.sub.15 together constitute an alkylene
bridge which together with the nitrogen atom between them forms a
five to seven-membered ring, [0258] (B) cyclohexyl, which is
optionally and independently substituted with: [0259] (i)
C.sub.1-3alkyl, [0260] (ii) hydroxyl; [0261] (c) --COR.sub.23,
wherein R.sub.23 is: [0262] (i) --NR.sub.24R.sub.25, wherein
R.sub.24 and R.sub.25 are each independently selected from: [0263]
(a) C.sub.1-6alkyl; [0264] or wherein R.sub.24 and R.sub.25
together constitute a alkylene bridge which together with the
nitrogen atom between them forms a five to six-membered ring,
wherein one of the methylene groups is optionally replaced by an
oxygen atom and which ring is optionally and independently
substituted with one or more of the following groups: [0265] (i)
hydroxyl, [0266] (ii) --CONH.sub.2, [0267] (D) is:
[0267] ##STR00007## [0268] which is optionally substituted with
halogen; [0269] (E) is selected from the following:
[0269] ##STR00008## [0270] R.sub.2 is: [0271] (A) CF.sub.3, [0272]
(B) cyano, [0273] (C) halogen, or [0274] (D) nitro, [0275] R.sub.3
is selected from the following:
[0275] ##STR00009## [0276] R.sub.4 and R.sub.5 are each
independently selected from: [0277] (A) hydrogen, [0278] (B)
C.sub.1-6alkyl, which is optionally and independently substituted
with one or more of the following groups: [0279] (i) oxo, [0280]
(ii) C.sub.3-5cycloalkyl, [0281] (iii) aryl or heteroaryl, each of
which is optionally and independently substituted with one or more
of the following groups: [0282] (a) C.sub.1-3alkyl which is
optionally substituted with fluorine, [0283] (b) --CO.sub.2H [0284]
(c) halogen, [0285] (iv) NH.sub.2 [0286] (v) hydroxyl, [0287] (vi)
--CONH.sub.2, [0288] (vii) fluorine, [0289] (viii) NHCOCH.sub.3
[0290] or R.sub.4 and R.sub.5 together constitute an alkylene
bridge which together with the nitrogen atom between them forms a
four- to seven-membered ring, wherein one of the methylene groups
is optionally replaced by an NH or oxygen atom and which ring is
optionally and independently substituted with one or more of the
following groups: [0291] (A) --CONH.sub.2, [0292] (B)
NR.sub.39R.sub.40 wherein R.sub.39 and R.sub.40 are optionally and
independently selected from: [0293] (i) C.sub.1-5alkylcarbonyl
[0294] (ii) C.sub.1-5alkylsulfonyl [0295] (iii) arylcarbonyl [0296]
(iv) arylsulfonyl [0297] (C) --OR.sub.44, wherein R.sub.44 is
selected from: [0298] (i) hydrogen, [0299] (ii)
C.sub.1-5alkylcarbonyl [0300] (D) oxo, or [0301] (E) fluorine
[0302] or wherein NR.sub.4R.sub.5 constitute a 5-membered
heteroaryl ring containing a total of 2 nitrogen hetero atoms in
the ring, or a tautomer, pharmaceutically acceptable salt, solvate
or amino-protected derivative thereof.
[0303] In yet another embodiment there are provided compounds are
compounds of formula (I) as described above and wherein: [0304] X
is --CH.sub.2--, --CH.sub.2CH.sub.2-- or --CH.sub.2CO--, [0305] Y
is --NH-- or --O--; [0306] R.sub.1 is: [0307] (A) phenyl, pyridyl,
naphthyl, quinolinyl or benzothienyl, each of which is optionally
and independently substituted with one or more of the following
groups: [0308] (i) C.sub.1-3alkyl, which is optionally substituted
with fluorine, [0309] (ii) C.sub.1-3alkoxy, which is optionally
substituted with fluorine, [0310] (iii) methylthio, which is
optionally substituted with fluorine, [0311] (iv) arylthio,
optionally substituted with NH.sub.2, [0312] (v) F, Cl or Br,
[0313] (vi) hydroxyl, [0314] (vii) NH.sub.2 or N(CH.sub.3).sub.2,
[0315] (viii) SO.sub.2CH.sub.3 [0316] (B) cyclohexyl, optionally
substituted with hydroxyl, [0317] R.sub.2 is: [0318] (A) CF.sub.3,
[0319] (B) cyano, [0320] (C) F, Cl, Br or [0321] (D) nitro, [0322]
R.sub.3 is:
[0322] ##STR00010## [0323] R.sub.4, R.sub.5 are each independently
selected from: [0324] (A) hydrogen, [0325] (B) C.sub.1-4alkyl,
which is optionally and independently substituted with one or more
of the following groups: [0326] (i) oxo, [0327] (ii) cyclopropyl,
[0328] (iii) aryl or heteroaryl selected from phenyl, pyridyl,
pyrimidyl, pyrazolyl and oxazolyl, each of which is optionally and
independently substituted with one or more of the following groups:
[0329] (a) C.sub.1-2alkyl, [0330] (b) fluorine or chlorine, [0331]
(iv) --CONH.sub.2 [0332] (v) hydroxyl, [0333] (vi) fluorine, [0334]
(vii) NHCOCH.sub.3 [0335] or R.sub.4 and R.sub.5 together
constitute an alkylene bridge which together with the nitrogen atom
between them forms a four- to seven-membered ring, wherein one of
the methylene groups is optionally replaced by an NH, and which
ring is optionally and independently substituted with one or more
of the following groups: [0336] (A) CONH.sub.2, [0337] (B)
hydroxyl, [0338] (C) C.sub.1-5alkylcarbonyloxy, [0339] (D) oxo,
[0340] (E) fluorine, [0341] (F) NR.sub.39R.sub.40 wherein R.sub.39
and R.sub.40 are optionally and independently selected from: [0342]
(i) hydrogen [0343] (ii) C.sub.1-5alkylcarbonyl [0344] (iii)
C.sub.1-5alkylsulfonyl [0345] (iv) Phenylcarbonyl, or [0346] (v)
phenylsulfonyl, [0347] or a tautomer, pharmaceutically acceptable
salt, solvate or amino-protected derivative thereof.
[0348] In a further embodiment there are provided compounds of
formula (I) as described above and wherein: [0349] X is
--CH.sub.2--, [0350] Y is --NH--, [0351] R.sub.1 is: [0352] (A)
phenyl or pyridyl optionally and independently substituted with one
or more of the following groups: [0353] (i) methyl, [0354] (ii)
CF.sub.3, [0355] (iii) OCF.sub.3, [0356] (iv) OCF.sub.2H [0357] (v)
OCH.sub.3, [0358] (vi) OCH(CH.sub.3).sub.2 [0359] (vii) SCF.sub.3,
[0360] (viii) arylthio substituted with NH.sub.2, [0361] (ix) F or
Cl, [0362] (x) N(CH.sub.3).sub.2, [0363] (xi) OCH.sub.2CF.sub.3
[0364] (xii) SO.sub.2CH.sub.3 [0365] (B) naphthyl, [0366] R.sub.2
is: [0367] (A) cyano, [0368] (B) Cl, or [0369] (C) nitro; [0370]
R.sub.3 is:
[0370] ##STR00011## [0371] R.sub.4 and R.sub.5 are each
independently selected from: [0372] (A) hydrogen, [0373] (B)
C.sub.1-3alkyl, optionally substituted with one or more of the
following groups: [0374] (i) hydroxyl, [0375] (ii) pyridyl, [0376]
(iii) 1-methyl-1H-pyrazole, [0377] (iv) 1,5-dimethyl-1H-pyrazole,
[0378] (v) --CONH.sub.2, or R.sub.4 and R.sub.5 together constitute
an alkylene bridge which together with the nitrogen atom between
them forms a four or five-membered ring, each optionally and
independently substituted with one or more of the following: [0379]
(A) hydroxyl, [0380] (B) amino [0381] (C) fluorine [0382] (D) oxo
[0383] (E) --NHSO.sub.2CH.sub.3 [0384] (F) --NHCOCH.sub.3 [0385] or
a tautomer, pharmaceutically acceptable salt, solvate or
amino-protected derivative thereof.
[0386] In still a further embodiment there are provided compounds
of formula (I) as described above and wherein: [0387] X is
--CH.sub.2--, [0388] Y is --NH--, [0389] R.sub.1 is: [0390] (A)
phenyl, optionally and independently substituted with one or more
of the following groups: [0391] (i) methyl, [0392] (ii) CF.sub.3,
[0393] (iii) OCF.sub.3, [0394] (iv) OCH.sub.3, [0395] (v)
SCF.sub.3, [0396] (vi) arylthio substituted with NH.sub.2, [0397]
(vii) F or Cl; [0398] (B) naphthyl, [0399] R.sub.2 is: [0400] (A)
cyano, [0401] (B) Cl, or [0402] (C) nitro; [0403] R.sub.3 is:
[0403] ##STR00012## [0404] R.sub.4 and R.sub.5 are each
independently selected from: [0405] (A) hydrogen, [0406] (B)
C.sub.1-2alkyl, optionally substituted with one or more of the
following groups: [0407] (i) hydroxyl, [0408] (ii) pyridyl, [0409]
(iii) 1,5-dimethyl-1H-pyrazole, [0410] (iv) --CONH.sub.2, [0411] or
R.sub.4 and R.sub.5 together constitute an alkylene bridge which
together with the nitrogen atom between them forms a four or
five-membered ring, each optionally and independently substituted
with one or more hydroxyl or oxo; or a tautomer, pharmaceutically
acceptable salt, solvate or amino-protected derivative thereof.
[0412] In yet a further embodiment there are provided the following
compounds:
##STR00013## ##STR00014## ##STR00015## ##STR00016## ##STR00017##
##STR00018## ##STR00019## ##STR00020## ##STR00021## ##STR00022##
##STR00023## ##STR00024## ##STR00025## ##STR00026## ##STR00027##
##STR00028## ##STR00029## ##STR00030## ##STR00031## ##STR00032##
##STR00033## ##STR00034## ##STR00035## ##STR00036## ##STR00037##
##STR00038## ##STR00039## ##STR00040## ##STR00041## ##STR00042##
##STR00043## ##STR00044## ##STR00045## ##STR00046## ##STR00047##
##STR00048## ##STR00049## ##STR00050## ##STR00051## ##STR00052##
##STR00053## ##STR00054## ##STR00055## ##STR00056## ##STR00057##
##STR00058## ##STR00059## ##STR00060## ##STR00061## ##STR00062##
##STR00063## ##STR00064## ##STR00065## ##STR00066## ##STR00067##
##STR00068## ##STR00069## ##STR00070## ##STR00071## ##STR00072##
##STR00073## ##STR00074## ##STR00075## ##STR00076## ##STR00077##
##STR00078## ##STR00079## ##STR00080## ##STR00081## ##STR00082##
##STR00083## ##STR00084## ##STR00085## ##STR00086##
[0413] In still an even further embodiment there are provided the
following compounds: [0414]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-fluoro-3-(trifluorom-
ethyl)benzyl]-5-nitropyrimidine-2,4-diamine; [0415]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-chloro-6-fluoro-3-me-
thylbenzyl)-5-nitropyrimidine-2,4-diamine; [0416]
N.sup.4-[(tans-4-aminocyclohexyl)methyl]-N.sup.2-(1-naphthylmethyl)-5-nit-
ropyrimidine-2,4-diamine; [0417]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-{2-[(trifluorom-
ethyl)thio]benzyl}pyrimidine-2,4-amine; [0418]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-chloro-N.sup.2-[2-(trifluorom-
ethoxy)benzyl]pyrimidine-2,4-diamine; [0419]
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)azetidin-3-one; [0420]
N.sup.4-{[trans-4-(3-aminoazetidin-1-yl)cyclohexyl]methyl}-5-nitro-N.sup.-
2-[2-(trifluoromethoxy)benzyl]-pyrimidine-2,4-diamine; [0421]
N-[1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-
-yl)amino]methyl}-cyclohexyl)azetidin-3-yl]methanesulfonamide;
[0422]
N-[1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-
-yl)amino]methyl}-cyclohexyl)azetidin-3-yl]acetamide; [0423]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]pyrimidine-2,4-diamine; [0424]
4-{[(trans-4-aminocyclohexyl)methyl]amino}-2-{[2-(trifluoromethoxy)benzyl-
]amino}pyrimidine-5-carbonitrile; [0425]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-{2-[(2-aminophenyl)thio-
]benzyl}-5-nitropyrimidine-2,4-diamine; [0426]
5-nitro-N.sup.4-[(trans-4-pyrrolidin-1-ylcyclohexyl)methyl]-N.sup.2-[2-(t-
rifluoromethoxy)benzyl]pyrimidine-2,4-diamine; [0427]
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)pyrrolidin-3-ol; [0428]
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-5-nitro-N.sup.2-[2-(tri-
fluoromethoxy)benzyl]pyrimidine-2,4-diamine; [0429]
N.sup.4-{[trans-4-(dimethylamino)cyclohexyl]methyl}-5-nitro-N.sup.2-[2-(t-
rifluoromethoxy)benzyl]pyrimidine-2,4-diamine; [0430]
4-({[trans-4-(dimethylamino)cyclohexyl]methyl}amino)-2-{[2-(trifluorometh-
oxy)benzyl]amino}-pyrimidine-5-carbonitrile; [0431]
5-nitro-N.sup.4-({trans-4-[(pyridin-3-ylmethyl)amino]cyclohexyl})methyl)--
N.sup.2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine; [0432]
5-nitro-N.sup.4-({trans-4-[(pyridin-4-ylmethyl)amino]cyclohexyl}methyl)-N-
.sup.2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine; [0433]
2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-y-
l)amino]methyl}cyclo-hexyl)amino]ethanol; [0434]
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)azetidin-3-ol; [0435]
5-nitro-N.sup.4-({trans-4-[(pyridin-2-ylmethyl)amino]cyclohexyl}methyl)-N-
.sup.2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine; [0436]
N.sup.2-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}-cyclohexyl)glycinamide; [0437]
N.sup.4-[(trans-4-{[(1,5-dimethyl-1H-pyrazol-4-yl)methyl]amino}cyclohexyl-
)methyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamin-
e; [0438]
N.sup.4-[(trans-4-{[(1-methyl-1H-pyrazol-4-yl)methyl]amino}cyclo-
hexyl)methyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-d-
iamine; [0439]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(2-isopropoxypyridin-3-
-yl)methyl]-5-nitropyrimidine-2,4-diamine; [0440]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-{[2-(2,2,2-trif-
luoroethoxy)pyridin-3-yl]methyl}pyrimidine-2,4-diamine;
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(5-fluoro-2-methoxypyr-
idin-3-yl)methyl]-5-nitropyrimidine-2,4-diamine; [0441]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(difluoromethoxy)ben-
zyl]-5-nitropyrimidine-2,4-diamine; [0442]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-chlorobenzyl)-5-nitr-
opyrimidine-2,4-diamine; [0443]
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-5-nitro-N.sup.2-{[2-(2,-
2,2-trifluoroethoxy)pyridin-3-yl]methyl}pyrimidine-2,4-diamine;
[0444]
N.sup.4-{[trans-4-(3-fluoroazetidin-1-yl)cyclohexyl]methyl}-5-nitro-N.sup-
.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine; [0445]
N.sup.4-{[trans-4-(ethylamino)cyclohexyl]methyl}-5-nitro-N.sup.2-[2-(trif-
luoromethoxy)benzyl]pyrimidine-2,4-diamine; [0446]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-chloro-3-(dimethylam-
ino)benzyl]-5-nitropyrimidine-2,4-diamine; [0447] 1-trans-4
{[(2-{[2-chloro-3-(dimethylamino)benzyl]amino}-5-nitropyrimidin-4-yl)amin-
o]methyl}cyclohexyl)azetidin-3-ol; [0448]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(5-chloro-2-methoxybenz-
yl)-5-nitropyrimidine-2,4-diamine; [0449]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-methoxybenzyl)-5-nit-
ropyrimidine-2,4-diamine; [0450]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[5-chloro-2-(trifluorom-
ethoxy)benzyl]-5-nitropyrimidine-2,4-diamine; [0451]
N.sup.2-[5-chloro-2-(trifluoromethoxy)benzyl]-5-nitro-N.sup.4-[(trans-4-p-
yrrolidin-1-ylcyclohexyl)-methyl]pyrimidine-2,4-diamine; [0452]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(2-methoxypyridin-3-yl-
)methyl]-5-nitropyrimidine-2,4-diamine; [0453]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-{[4-(trifluorom-
ethyl)pyridin-3-yl]methyl}pyrimidine-2,4-diamine; [0454]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[5-chloro-2-(methylsulf-
onyl)benzyl]-5-nitropyrimidine-2,4-diamine; [0455]
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-N.sup.2-[(4-chloropyrid-
in-3-yl)methyl]-5-nitropyrimidine-2,4-diamine; [0456]
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-5-nitro-N.sup.2-{[4-(tr-
ifluoromethyl)pyridin-3-yl]methyl}pyrimidine-2,4-diamine; [0457]
1-[trans-4-({[5-nitro-2-{[4-(trifluoromethyl)pyridin-3-yl]methyl}amino)py-
rimidin-4-yl]amino}methyl)cyclohexyl]azetidin-3-ol; [0458]
(2R)-1-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]propan-2-ol; [0459]
(2S)-1-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]propan-2-ol; [0460]
3-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-y-
l)amino]methyl}cyclohexyl)amino]propan-1-ol; [0461]
(2R)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]propan-1-ol [0462]
(2S)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]propan-1-ol;
General Synthetic Methods
[0463] The compounds of the invention may be prepared by the
methods described below. In each of the schemes below, the groups
R.sub.1, R.sub.2 and R.sub.3 are as defined above for general
formula I unless noted otherwise. Optimum reaction conditions and
reaction times may vary depending on the particular reactants used.
Unless otherwise specified, solvents, temperatures, pressures and
other reaction conditions may be readily selected by one of
ordinary skill in the art. Specific procedures are provided in the
Synthetic Examples section. Typically, reaction progress may be
monitored by thin layer chromatography (TLC) if desired.
Intermediates and products may be purified by chromatography on
silica gel and/or recrystallization. Starting materials and
reagents are either commercially available or may be prepared by
one skilled in the art from commercially available materials using
methods described in the chemical literature.
[0464] Compounds of formula (I) may be prepared as illustrated in
Scheme I and described below.
##STR00087##
[0465] As illustrated above, a 2,4-dihalopyrimidine (II),
preferably a 2,4-dichloropyrimidine, is reacted with about one
equivalent of an amine (R'R''NH) in the presence of a base, such as
triethylamine, in a suitable solvent, such as EtOH, to provide
intermediate III. The reaction is carried out preferably at about
0.degree. C. to about room temperature. Intermediate III is then
reacted with a second amine R.sub.1XNH.sub.2 in a suitable solvent,
such as EtOH, to provide the desired I. The reaction is preferably
heated to about the reflux temperature of the solvent. For
intermediates III having R.sub.2 groups that are less electron
withdrawing than NO.sub.2, such as R.sub.2=F, Cl, CN or CO.sub.2Et,
the reaction is preferably carried out in a sealed vessel in a
microwave reactor at about 140.degree. C.
[0466] If R.sub.3 contains a second amine group, (i.e., in the R'
and/or R'' groups in Scheme I above) the second amine is preferably
protected with a suitable amino-protecting group, for example with
a Boc-group, prior to reaction with intermediate H, and the amine
is deprotected after reaction of the pyrimidine intermediate III
with R.sub.1XNH.sub.2. For example, in the case of 1-amino
4-aminomethylcyclohexane as illustrated in Scheme II, the
mono-Boc-protected diamine is reacted with II as described above.
The resulting intermediate IV is then reacted with R.sub.1XNH.sub.2
as described above, and the Boc-protected intermediate V is then
deprotected by treatment with acid to provide the desired compound
of formula (I). The free amino group is then reacted with suitable
reagents, such as alkylating agents or, under reductive conditions,
carbonyl compounds, to provide the N-monoalkylated or N-dialkylated
product of formula (I)
##STR00088##
[0467] in a variation illustrated in Scheme III, if R.sub.2 is
NO.sub.2, intermediate II may be reacted with a thiocyanate salt,
such as potassium thiocyanate, in a suitable solvent, such as EtOH,
to produce VI. Intermediate VI is reacted with R.sub.1XNH.sub.2 in
a suitable solvent, such as EtOH, and in the presence of a base,
such as triethylamine, to provide VII. Intermediate VII may then be
reacted with an amine R'R''NH in a suitable solvent, such as EtOH
or methylene chloride, to provide the desired compound of formula
I
##STR00089##
[0468] Substituents R.sub.1, R.sub.2 and R.sub.3 may be further
modified by methods known in the art to obtain additional compounds
of formula (I). Some of these modifications are illustrated in the
synthetic examples below.
[0469] Compounds of formula (I) having Y=O or S may be prepared by
reacting VII with the desired R'OH or R'SH in the presence of a
suitable base such as sodium hydride in a suitable solvent such as
THF or DMF to obtain I (Y--R.sub.3=--OR or --SR' respectively).
[0470] In order that this invention be more fully understood, the
following examples are set forth. These examples are for the
purpose of illustrating embodiments of this invention, and are not
to be construed as limiting the scope of the invention in any way.
Starting materials used are either commercially available or easily
prepared from commercially available materials by those skilled in
the art.
EXPERIMENTAL SECTION
Example 1
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(cyclohexylmethyl)-5-nit-
ropyrimidine-2,4-diamine
##STR00090##
[0472] To a solution of 2,4-dichloro-5-nitropyrimidine (1.77 g,
9.11 mmol) in a mixture of AcCN (75 mL) and DMF (8 mL), were added
tert-butyl trans-4-aminomethylcyclohexylcarbamate (2.08 g, 9.11
mmol) and diisopropylethylamine (DIPEA) (1.59 mL, 9.11 mmol). The
reaction mixture was stirred at room temperature for 1 h. The
reaction mixture was diluted with EtOAc and washed with water
(.times.4). The organic phase was dried over anhydrous
Na.sub.2SO.sub.4 and concentrated in vacuo. The resulting residue
was purified by silica gel column chromatography using 9:1
EtOAc:hexanes as an eluent to afford 2.21 g (63%) of the desired
product as a yellowish solid.
[0473] To a solution of the above
{4-[(2-chloro-5-nitro-pyrimidinylamino)-methyl]-cyclohexyl}-carbamic
acid tert-butyl ester (50.2 mg, 0.13 mmol) in CH.sub.2Cl.sub.2 (0.8
mL) was added cyclohexylmethylamine (44.1 mg, 0.39 mmol). The
reaction mixture was stirred at room temperature for 3 h and then
purified by silica gel column chromatography using 50:1
CH.sub.2Cl.sub.2;MeOH as an eluent to afford 32 mg (54%) of the
desired product as a pale yellow solid.
[0474] The above
(4-{[2-(cyclohexylmethylamino)-5-nitro-pyrimidin-4-ylamino]-methyl}-cyclo-
hexyl)-carbamic acid tert-butyl ester (32 mg, 0.07 mmol) was
dissolved in dioxane (0.6 mL) and treated with 4M HCl in dioxane
(0.25 mL). The reaction mixture was concentrated in vacuo and the
resulting residue was purified by silica gel preparative TLC to
afford 9 mg (35%) of the title compound, m/z 363.4 [M+1].sup.+.
[0475] The following compounds were prepared using similar
procedures as described above: [0476]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(5-fluoro-2-methylbenzy-
l)-5-nitropyrimidine-2,4-diamine, m/z 389.4 [M+1].sup.+ [0477]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-methoxybenzyl)-5-nit-
ropyrimidine-2,4-diamine, m/z 387.4 [M+1].sup.+ [0478]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N2-(4-chloro-2-methylbenzyl)-5--
nitropyrimidine-2,4-diamine, m/z 405.4 [M+1].sup.+ [0479]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-fluoro-3-trifluorome-
thyl)benzyl]-5-nitropyrimidine-2,4-diamine, m/z 443.6 [M+1].sup.+
[0480]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-chloro-6-fluoro-3-me-
thylbenzyl)-5-nitropyrimidine-2,4-diamine, m/z 423.6 [M+1].sup.+
[0481]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(1-naphthylmethyl)-5-ni-
tropyrimidine-2,4-diamine, m/z 407.6 [M+1].sup.+ [0482]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-{2-[(trifluorom-
ethyl)thio]benzyl}pyrimidine-2,4-diamine, m/z 457.6 [M+1].sup.+
[0483]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(2-fluorophenyl)ethy-
l]-5-nitropyrimidine-2,4-diamine, m/z 389.8 [M+1].sup.+. [0484]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N-2-[2-(3-fluorophenyl)ethyl]-5-
-nitropyrimidine-2,4-diamine, m/z 389.8 [M+1].sup.+. [0485]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(4-fluorophenyl)ethy-
l]-5-nitropyrimidine-2,4-diamine, m/z 389.8 [M+1].sup.+. [0486]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(1H-indol-3-yl)ethyl-
]-5-nitropyrimidine-2,4-diamine, m/z 408.7 [M-1].sup.+. [0487]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(2-pyridin-2-yl-
ethyl)pyrimidine-2,4-diamine, m/z 372.8 [M+1].sup.+. [0488]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(2-pyridin-3-yl-
ethyl)pyrimidine-2,4-diamine, m/z 372.8 [M+1].sup.+. [0489]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(2-pyridin-4-yl-
ethyl)pyrimidine-2,4-diamine, m/z 372.9 [M+1].sup.+. [0490]
N.sup.2-(4-aminobenzyl)-N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-
pyrimidine-2,4-diamine, m/z 372.9 [M+1].sup.+. [0491]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(pyridin-4-ylme-
thyl)pyrimidine-2,4-diamine, m/z 358.9 [M+1].sup.+. [0492]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(quinolin-4-ylm-
ethyl)pyrimidine-2,4-diamine, m/z 408.3 [M+1].sup.+. [0493]
4-{[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)ami-
no]methyl}benzene-1,2-diol, m/z 389.6 [M+1].sup.+. [0494]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(1,3-benzodioxol-5-ylme-
thyl)-5-nitropyrimidine-2,4-diamine, m/z 401.6 [M+1].sup.+. [0495]
4-{[(4-{[(trans-4-aminocyclohexyl)methyl]amino)}-5-nitropyrimidin-2-yl)am-
ino]methyl}-2-methoxyphenol, m/z 403.4 [M+1].sup.+. [0496]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(1H-benzimidazol-2-y-
l)ethyl]-5-nitropyrimidine-2,4-diamine, m/z 411.6 [M+1].sup.+.
[0497]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(1-benzothien-3-ylmethy-
l)-5-nitropyrimidine-2,4-diamine, m/z 413.3 [M+1].sup.+. [0498]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(5-chloro-1-benzothien-
-3-yl)methyl]-5-nitropyrimidine-2,4-diamine, m/z 447.3 [M+1].sup.+.
[0499]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[4-(dimethylamino)benzy-
l]-5-nitropyrimidine-2,4-diamine, m/z 398.4 [M-1].sup.+. [0500]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(difluoromethoxy)ben-
zyl]-5-nitropyrimidine-2,4-diamine, m/z 423.4 [M+1].sup.+. [0501]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-chlorobenzyl)-5-nitr-
opyrimidine-2,4-diamine, m/z 391.2 [M+1].sup.+. [0502]
N.sup.2-(4-({[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-y-
l-N,N-dimethylglycinamide, m/z 352.6 [M+1].sup.+. [0503]
N.sup.2-(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl-
)-N-methylglycinamide, m/z 338.5 [M+1].sup.+. [0504] Methyl
N-(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)glyci-
nate, m/z 339.4 [M+1].sup.+. [0505] methyl
N-(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)-L-al-
aninate, m/z 353.4 [M+1].sup.+. [0506] Methyl
(2S)-[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)a-
mino](phenyl)acetate, m/z 415.4 [M+1].sup.+. [0507] Methyl
(2R)-[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)a-
mino](phenyl)acetate, m/z 415.4 [M+1].sup.+. [0508]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(3-bromobenzyl)-5-nitro-
pyrimidine-2,4-diamine, m/z 435.5 [M+1].sup.+. [0509]
N.sup.4-(4-Amino-cyclohexylmethyl)-N.sup.2-(3-bromo-2-methyl-benzyl)-5-ni-
tro-pyrimidine-2,4-diamine, m/z 450.4 [M+1].sup.+ [0510]
N.sup.2-(5-amino-2-chlorobenzyl)-N.sup.4-[(trans-4-aminocyclohexyl)methyl-
]-5-nitropyrimidine-2,4-diamine, m/z 406.4[M+1].sup.+ [0511]
N.sup.2-(3-aminobenzyl)-N4-[(trans-4-aminocyclohexyl)methyl]-5-nitropyrim-
idine-2,4-diamine, m/z 372.5 [M+1].sup.+ [0512]
N-(3-{[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)-
amino]methyl}phenyl)acetamide, m/z 414.6 [M+1].sup.+ [0513]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[3-(dimethylamino)benzy-
l]-5-nitropyrimidine-2,4-diamine, m/z 400.5 [M+1].sup.+ [0514]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-benzyl-5-nitropyrimidin-
e-2,4-diamine, m/z 357.6 [M+1].sup.+ [0515]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(dimethylamino)benzy-
l]-5-nitropyrimidine-2,4-diamine, m/z 371.6 [M+1].sup.+ [0516]
N-(3-{[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)-
amino]methyl}phenyl)-N-methylacetamide, m/z 428.6 [M+1].sup.+
[0517]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2,3-dihydro-1-benzofur-
an-5-ylmethyl)-5-nitropyrimidine-2,4-diamine, m/z 399.5 [M+1].sup.+
[0518]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[3-methylamino)benzyl]--
5-nitropyrimidine-2,4-diamine, m/z 386.7 [M+1].sup.+ [0519]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-methylbenzyl)-5-nitr-
opyrimidine-2,4-diamine, m/z 371.6 [M+1].sup.+ [0520]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-fluorobenzyl)-5-nitr-
opyrimidine-2,4-diamine, m/z 375.6 [M+1].sup.+ [0521]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(3-methylbenzyl)-5-nitr-
opyrimidine-2,4-diamine, m/z 371.7 [M+1].sup.+ [0522]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(3-chlorobenzyl)-5-nitr-
opyrimidine-2,4-diamine, m/z 390.8 [M+1].sup.+ [0523]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(3-fluorobenzyl)-5-nitr-
opyrimidine-2,4-diamine, m/z 375.6 [M+1].sup.+ [0524]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[3-(trifluorome-
thoxy)benzyl]pyrimidine-2,4-diamine, m/z 441.6 [M+1].sup.+ [0525]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2,3-dimethylbenzyl)-5--
nitropyrimidine-2,4-diamine, m/z 385.7 [M+1].sup.+ [0526]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(3-chloro-2-methylbenzy-
l)-5-nitropyrimidine-2,4-diamine, m/z 405.6 [M+1].sup.+ [0527]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(3-chloro-2-fluorobenzy-
l)-5-nitropyrimidine-2,4-diamine, m/z 409.6 [M+1].sup.+ [0528]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2,3-difluorobenzyl)-5--
nitropyrimidine-2,4-diamine, m/z 393.6 [M+1].sup.+ [0529]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(5-chloro-2-fluorobenzy-
l)-5-nitropyrimidine-2,4-diamine m/z 410.5 [M+1].sup.+ [0530]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(5-chloro-2-methylbenzy-
l)-5-nitropyrimidine-2,4-diamine m/z 407.2 [M+1].sup.+ [0531]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(3,4-difluorobenzyl)-5--
nitropyrimidine-2,4-diamine, m/z 392.9 [M+1].sup.+ [0532]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(3-chloro-4-fluorobenzy-
l)-5-nitropyrimidine-2,4-diamine, m/z 410.0 [M+1].sup.+ [0533]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(3,5-difluorobenzyl)-5--
nitropyrimidine-2,4-diamine, m/z 392.9 [M+1].sup.+ [0534]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-chloro-4-fluorobenzy-
l)-5-nitropyrimidine-2,4-diamine, m/z 410.3 [M+1].sup.+ [0535]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(3,5-dichlorobenzyl)-5--
nitropyrimidine-2,4-diamine, m/z 425.5 [M+1].sup.+ [0536]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(4-chloro-2-fluorobenzy-
l)-5-nitropyrimidine-2,4-diamine, m/z 409.6 [M+1].sup.+ [0537]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(4-bromo-2-fluorobenzyl-
)-5-nitropyrimidine-2,4-diamine, m/z 455.5 [M+1].sup.+ [0538]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[4-fluoro-2-(trifluorom-
ethyl)benzyl]-5-nitropyrimidine-2,4-diamine, m/z 443.6 [M+1].sup.+
[0539]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[4-fluoro-3-(trifluorom-
ethyl)benzyl]-5-nitropyrimidine-2,4-diamine, m/z 443.6 [M+1].sup.+
[0540]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-chloro-5-(dimethylam-
ino)benzyl]-5-nitropyrimidine-2,4-diamine, m/z 434.5 [M+1].sup.+
[0541]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-chloro-3-(dimethylam-
ino)benzyl]-5-nitropyrimidine-2,4-diamine, m/z 434.8 [M+1].sup.+
[0542]
3-{[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)ami-
no]methyl}benzamide, m/z 400.4 [M+1].sup.+ [0543]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(pyridin-3-ylme-
thyl)pyrimidine-2,4-diamine m/z 358.5 [M+1].sup.+. [0544]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-{[6-(trifluorom-
ethyl)pyridin-3-yl]methyl}pyrimidine-2,4-diamine m/z 426.5
[M+1].sup.+ [0545]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(3-chloropyridi-
n-2-yl)methyl]-5-nitropyrimidine-2,4-diamine m/z 392.4 [M+1].sup.+
[0546]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(2-methoxypyridin-3-yl-
)methyl]-5-nitropyrimidine-2,4-diamine m/z 388.5 [M+1].sup.+ [0547]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-{[2-(trifluorom-
ethyl)pyridin-3-yl]methyl}pyrimidine-2,4-diamine m/z 426.4
[M+1].sup.+ [0548]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(4,5-dichloropy-
ridin-3-yl)methyl]-5-nitropyrimidine-2,4-diamine m/z 426.5
[M+1].sup.+ [0549]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(3-methoxypyrid-
in-2-yl)methyl]-5-nitropyrimidine-2,4-diamine m/z 388.7 [M+1].sup.+
[0550]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(pyrazin-2-ylme-
thyl)pyrimidine-2,4-diamine, m/z 359 [M+1].sup.+. [0551]
(3-Aminomethyl-2-chloro-phenyl)-dimethyl-amine intermediate was
synthesized according to the following procedure:
##STR00091##
[0552] A 100 mL round bottomed flask was charged with
2-chloro-3-aminobenzoic acid (1.00 g, 5.38 mmol) methanol (50 mL)
and sulfuric acid (0.1 mL). The reaction was heated to reflux for
12 h. The reaction was cooled, and the solvent was removed. The
residual material was taken up in ethyl acetate (35 mL) and washed
with water (4.times.20 mL). The organics were dried (MgSO4),
concentrated, then purified by column chromatography using
hexanes/ethyl acetate (4:1) to yield 2-chloro-3-amino-benzoic acid
methyl ester (990 mg, 86%)
[0553] 2-Chloro-3-amino-benzoic acid methyl ester (1.20 g, 6.47
mmol) was placed in a round bottomed flask with DMF (5 mL),
Potassium carbonate (1.97 g, 14.2 mmol) and methyl iodide (0.89 mL,
14.22 mmol) were added. A reflux condenser was fitted and the
reaction was heated to 60.degree. C. for 12 h. The reaction was
cooled to rt, diluted with water (20 mL) and extracted with ethyl
acetate (3.times.20 mL). The organics were dried (MgSO4),
concentrated, then purified by column chromatography using
hexanes/ethyl acetate (1:1) to afford
2-chloro-3-dimethylamino-benzoic acid methyl ester (1.04 g,
75%)
[0554] 2-Chloro-3-dimethylamino-benzoic acid methyl ester (0.85 g,
3.98 mmol) was placed in a round bottomed flask with
tetrahydrofuran (50 mL) and cooled to 0.degree. C. Lithium aluminum
hydride (0.30 g, 7.96 mmol) was added in portions and after all the
LAH was added, the reaction was allowed to warm and stir at rt for
14. The reaction was quenched by the addition of ice, then water
(10 mL) was added. The organics were extracted into ethyl acetate
(3.times.15 mL), dried (MgSO4) and concentrated down to afford
(2-chloro-3-dimethylamino-phenyl)methanol which was taken onward
without further purification.
[0555] (2-Chloro-3-dimethylamino-phenyl)-methanol (0.50 g, 2.69
mmol) was placed in a round bottomed flask with dichloromethane (10
mL) and cooled to 0.degree. C. Triethylamine (0.68 g, 6.73 mmol),
methanesulfonylchloride (0.23 mL, 2.97 mmol), and
dimethylaminopyridine (0.02 g) were added and the reaction was
allowed to warm and stir at rt over 3 h. The reaction was diluted
with dichloromethane (20 mL) and washed with 1N Hydrochloric acid
(10 mL), 10% aqueous sodium bicarbonate (10 mL), and water (10 mL).
The organics were dried (MgSO4) and concentrated to afford
methanesulfonic acid 2-chloro-3-dimethylamino-benzyl ester.
[0556] Methanesulfonic acid 2-chloro-3-dimethylamino-benzyl ester
was placed in a round bottomed flask with DMF (8 mL). Sodium azide
(0.35 g, 5.38 mmol) was added and the reaction was heated to
60.degree. C. for 3 h. The reaction was cooled to rt, and the
solvent was removed in vacuo. The residual crude material was
purified by column chromatography using hexanes/ethyl acetate (1:1)
as eluent to afford
(3-azidomethyl-2-chloro-phenyl)-dimethyl-amine
[0557] (3-Azidomethyl-2-chloro-phenyl)-dimethyl-amine (0.25 g, 1.19
mmol) was placed in a round bottomed flask with tetrahydrofuran (25
mL). Triphenylphosphine (0.78 g, 2.97 mmol) was added and the
reaction was allowed to stir for 3 h at rt. The resulting
precipitate was filtered off and set aside. The filtrate was
concentrated down, then purified by column chromatography using
methanol/methylene chloride/ammonium hydroxide (5:95:0.1) to afford
the desired intermediate.
[0558] The following amine intermediates were prepared using
similar procedures as described above: [0559]
(3-Aminomethyl-4-chloro-phenyl)-dimethylamine [0560]
(2-Methoxy-pyridin-3-yl)-methylamine [0561]
(2-trifluoromethyl-pyridin-3-yl)-methylamine [0562]
(4,5-Dichloro-pyridin-3-yl)-methylamine [0563]
(3-Methoxy-pyridin-2-yl)-methylamine [0564]
Pyrazin-2-yl-methylamine [0565] (2-Methoxy-pyridin-3-yl)-methanol
intermediate was prepared according to the following procedure:
##STR00092##
[0566] To a stirred solution of 1.7M t-butyl lithium in THF (35.0
mL, 59.6 mmol, 2.60 equiv.) with THF (150 mL) at -78 degrees was
added dropwise 2-bromomesitylene (4.56 mL, 29.8 mmol, 1.30 equiv.).
After stirring for 1 h, 2-methoxy-pyridine (2.5 g, 22.9 mmol) was
added dropwise. This solution was warmed to -23 degrees and stirred
for 3 h and then cooled to -78 degrees again. DMF (2.66 mL, 34.4
mmol, 1.5 equiv.) was added, the solution was stirred at -78
degrees for 1 h. The reaction was quenched at this temperature with
brine (150 mL) and extracted with ether. The combined ether was
dried over K.sub.2CO.sub.3 and evaporated on vacuo. The residue was
chromatographed with 50%-80% EtOAc/Hexanes to give
2-methoxy-pyridine-3-carbaldehyde (2.34 g, 17.1 mmol, 74.5%) as a
yellow solid.
[0567] To a solution of 2-methoxy-pyridine-3-carbaldehyde (2.05 g,
14.9 mmol) in MeOH (70 mL) cooled to 0.degree. C. was added sodium
borohydride (670 mg, 17.7 mmol, 1.18 equiv.) as a solid in one
portion. The reaction was allowed to slowly warm to room
temperature and stirred for 2 h. Excess hydride was consumed by the
addition of H.sub.2O and the reaction mixture was concentrated
under reduced pressure. The residue was taken back up in EtOAc and
washed with H.sub.2O. The aqueous phase was back extracted with
EtOAc and the combine organic phase was washed with brine, dried
over anhydrous Na.sub.2SO.sub.4 and concentrated under reduced
pressure. The crude residue was purified by flash silica gel
chromatography using a 5% MeOH/DCM to provide, after concentration
of the eluent, to give (2-methoxy-pyridin-3-yl)-methanol (1.65 g,
11.9 mmol, 81.3%) as a white solid.
[0568] The following intermediates were made using similar
procedures described above: [0569]
(2-Trifluoromethyl-pyridin-3-yl)-methanol [0570]
(4,5-Dichloro-pyridin-3-yl)-methanol [0571]
(3-Methoxy-pyridin-2-yl)methanol
Example 2
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluoromet-
hoxy)benzyl]pyrimidine-2,4-diamine
##STR00093##
[0573] 2,4-Dichloro-5-nitropyrimidine (12 g) was dissolved in AcOH
(70 mL) and cooled to 0.degree. C. Potassium thiocyanate (6.31 g)
was added portion-wise to the solution over 2 h. The reaction was
diluted with water and filtered. The filter cake was washed with
cold ether to afford the desired
2-chloro-5-nitro-4-thiocyanatopyrimidine (9.5 g, 71%).
[0574] To a solution of 2-chloro-5-nitro-4-thiocyanatopyrimidine (1
g, 4.6 mmol) in DMF (5 mL) was added
2-(trifluoromethoxy)benzylamine (883 mg, 4.6 mmol) followed by
DIPEA (804 .mu.L, 4.6 mmol). The reaction mixture was stirred at
room temperature for 30 min and diluted with EtOAc. The solution
was washed with water (.times.4). The organic phase was dried over
anhydrous Na.sub.2SO.sub.4 an then concentrated in vacuo. The
resulting residue was purified by silica gel prep TLC using
CH.sub.2Cl.sub.2 as an eluent to afford 727 mg (42%) of the desired
product as a yellowish solid.
[0575] To a solution of the above product (50 mg, 0.135 mmol) in
DMF (1 mL) were added tert-butyl
trans-4-aminomethylcyclohexylcarbamate (61 mg, 0.27 mmol) and DIPEA
(47 mL, 0.27 mmol). The reaction mixture was stirred at room
temperature for 2 h. The reaction mixture was diluted with EtOAc
(20 mL) and washed with water (.times.3). The organic phase was
dried over anhydrous Na.sub.2SO4 and concentrated in vacuo. The
resulting residue was purified by silica gel prep TLC using 95:5
CH.sub.2Cl.sub.2:MeOH as an eluent to afford the 71.8 mg (99%) of
the desired product.
[0576] The above substituted diamino product (70 mg, 0.13 mmol) was
dissolved in CH.sub.2Cl.sub.2 (10 mL) and TFA (0.7 mL) was added.
The reaction mixture was stirred at room temperature for 40 min.
until all the starting material was consumed. The mixture was then
treated with a mixture of saturated NaHCO.sub.3 and 1M
Na.sub.2CO.sub.3 until pH 9-10. The mixture was extracted with
CH.sub.2Cl.sub.2 (.times.2). MeOH was added to the organic phase
during work-up to solubilize the product. The combined organic
phase was dried over anhydrous Na.sub.2SO.sub.4 and concentrated in
vacuo. The resulting residue was purified by silica gel preparative
TLC using CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH (10:1:0.1) as an eluent
to afford 39 mg (69%) of the title compound, m/z 438.8
[M-1].sup.+.
[0577] The following compound was prepared using similar procedures
as described above: [0578]
N.sup.4-[(cis-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorometh-
oxy)benzyl]pyrimidine-2,4-diamine, m/z 441.3 [M+1].sup.+
Example 3
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(4-chloropyridin-3-yl)m-
ethyl]-5-nitropyrimidine-2,4-diamine
##STR00094##
[0580] To a solution of
(4-[(2-chloro-5-nitro-pyrimidin-4-ylamino)-methyl]cyclohexyl)-carbamic
acid tert-butyl ester (48.2 mg, 0.13 mmol) in CH.sub.2Cl.sub.2 (0.8
mL) was added 3-aminomethyl-4-chloropyridine (53.5 mg, 0.38 mmol).
The reaction mixture was stirred at room temperature for 3 h. The
mixture was then purified by silica gel column chromatography using
50:1 CH.sub.2Cl.sub.2:MeOH as an eluent to afford 20.6 mg (34%) of
the desired product as a pale yellow solid.
[0581] The above
[4-({2-[(4-chloro-pyridin-3-ylmethyl)-amino]-5-nitro-pyrimidin-4-ylamino}-
-methyl)-cyclohexyl]-carbamic acid tert-butyl ester (20.6 mg, 0.042
mmol) was dissolved in dioxane (0.5 mL) and 4 M HCl in dioxane (200
.mu.L) was added to the solution. The reaction mixture was stirred
at room temperature for 4 h and concentrated in vacuo. The
resulting residue was purified by silica gel column chromatography
using a mixture of 10% 2M NH.sub.3 in MeOH and CH.sub.2Cl.sub.2 as
an eluent to afford 5.5 mg (34%) of the desired product, m/z 392.3
[M+1].sup.+.
Preparation of 3-aminomethyl-4-chloropyridine intermediate
[0582] To a solution of 4-chloro-nicotinic acid (2.5 g, 15.9 mmol)
in DMF (26 mL)), were added dicyclohexylcarbodiimide (DCC) (7.75 g,
37.5 mmol), 4-dimethylaminopyridine (DMAP) (0.286 g, 2.1 mmol) and
EtOH (2.6 mL, 46.8 mmol). The reaction mixture was stirred at room
temperature for 4 h. The mixture was then distilled (0.1 mmHg) to
afford 1.5 g (51%) of 4-chloro-nicotinic acid ethyl ester as a
colorless oil.
[0583] To a solution of 4-chloro-nicotinic acid ethyl ester (1.5 g,
8.1 mmol) in ether (15 mL) was added lithium aluminum hydride (0.31
g, 8.2 mmol) in ether (10 mL) over 10 min while cooling. The
reaction mixture was stirred for 1 h and quenched with water (2 mL)
and 15% NaOH (0.5 mL). The reaction mixture was filtered and the
residue on the filter was washed with ether. The combined filtrate
was dried over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo
to afford 0.75 g of (4-chloro-pyridin-3-yl)-MeOH as a yellowish
solid.
[0584] 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) (0.69 mL, 4.5 mmol)
was added to a solution of the above (4-chloro-pyridin-3-yl)-MeOH
(0.51 g, 3.6 mmol) and diphenylphosphoryl azide (DPPA) (0.92 mL,
4.3 mmol) in toluene (8 mL). The reaction mixture was stiffed at
room temperature for 3 b and then concentrated in vacuo. The
resulting residue was purified by silica gel column chromatography
using 1:4 EtOAc:hexanes to afford 0.47 g (79%) of
3-azidomethylchloropyridine.
[0585] Triphenylphosphine (0.81 g, 3.1 mmol) was added to a
solution of the above 3-azidomethyl-4-chloro-pyridine (0.47 g, 3.1
mmol) in anhydrous THF (10 mL) at 0.degree. C. The reaction mixture
was warned to room temperature and stirred for 18 h. The reaction
mixture was then diluted with NH.sub.4OH (3 mL) and stirred for
another 3 h. 3M NaOH was then added to the reaction mixture and
stirred for 1 h. The mixture was acidified to pH 2 by adding 4M HCl
solution. The mixture was diluted with ether and the layers were
separated. The aqueous layer was extracted with ether and then
basified with 2M NaOH solution. The aqueous solution was extracted
with CH.sub.2Cl.sub.2 (.times.3). The combined organic phase was
dried over MgSO.sub.4 and concentrated in vacuo to afford a pale
yellow oily solid. This product was diluted with ether and
filtered. The filtrated was concentrated in vacuo to afford a pale
yellow oil. The crude product was purified by silica gel column
chromatography using 20:1 CH.sub.2Cl.sub.2:MeOH as an eluent to
afford 0.21 g (53%) of 3-aminomethyl-4-chloropyridine as a
colorless oil.
[0586] The following compounds were prepared using similar
procedures as described above: [0587]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(2-chloropyridin-3-yl)-
methyl]-5-nitropyrimidine-2,4-diamine, m/z 392.4 [M+1].sup.+
Preparation of 3-aminomethyl-2-chloropyridine intermediate
[0588] A mixture of 2-chloronicotinic acid (5.0 g, 31.7 mmol) and
thionyl chloride (20 mL) was heated at reflux for 1.5 h. The
reaction mixture was concentrated in vacuo to remove an excess
thionyl chloride. The resulting solid was added in portions to a
solution of sodium borohydride (4.3 g, 114 mmol) in water at
10.degree. C. The reaction was maintained at 10-15.degree. C.
during the addition to NaBH.sub.4. The reaction mixture was then
warmed to room temperature. The mixture was saturated with NaCl and
extracted with ether (30 mL.times.3). The combined organic layer
was dried over MgSO.sub.4 and concentrated in vacuo to afford 4.1 g
(90%) of (2-chloro-pyridin-3-yl)-methanol.
[0589] To a solution of the above (2-chloro-pyridin-3-yl)-methanol
(2.2 g, 15.1 mmol) and DPPA (3.9 mL, 18.2 mmol) in toluene (30 mL)
was added DBU (2.9 mL, 18.2 mmol) at room temperature and stirred
for 3 h. The reaction was diluted with 3M HCl (30 mL) and Et.sub.2O
(75 mL). The layers were separated and the aqueous layer was
extracted with Et.sub.2O (75 mL). The combined organic layers were
washed with water (50 ml), brine and dried over MgSO.sub.4. The
solution was filtered and concentrated in vacuo to yield a yellow
oil. Silica gel column chromatography using 95:5 hexanes:EtOAc
afforded 2.0 g (77%) of 3-azidomethyl-2-chloropyridine as a
colorless oil.
[0590] Triphenylphosphine (2.5 g, 9.5 mmol) was added to a solution
of the above 3-azidomethyl-2-chloro-pyridine (1.4 g, 8.5 mmol) in
anhydrous THF (30 mL) at 0.degree. C. The reaction mixture was
warned to room temperature and stirred for 18 h. The reaction
mixture was diluted with NH.sub.4OH (3 mL) and stirred for another
3 h. The mixture was then treated with 3M NaOH and stirred for 1 h.
The mixture was acidified to pH 2 by adding 4M HCl solution. The
mixture was diluted with ether and the layers were separated. The
aqueous layer was extracted with ether and then basified with 2M
NaOH solution. The aqueous solution was extracted with
CH.sub.2Cl.sub.2 (.times.3). The combined organic phase was dried
over MgSO.sub.4 and concentrated in vacuo to afford a pale yellow
oily solid. This product was diluted with ether and filtered. The
filtrated was concentrated to afford a pale yellow oil. The crude
product was purified by silica gel column chromatography using 20:1
CH.sub.2Cl.sub.2:MeOH as an eluent to afford 0.88 g (73%) of
3-aminomethyl-2-chloropyridine as a colorless oil. [0591]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(5-chloro-2-methoxybenz-
yl)-5-nitropyrimidine-2,4-diamine, m/z 421.3 [M+1].sup.+
Preparation of 5-chloro-2-methoxy-benzylamine intermediate
[0592] A mixture of methyl 5-chloro-2-methoxybenzoate (2.5 g, 12.5
mmol) and 7 M NH.sub.3 (25 mL) in MeOH was heated to 120.degree. C.
in a sealed tube for 72 h. The reaction mixture was cooled to room
temperature and nitrogen was bubbled through the solution for 15
min. The solution was then concentrated in vacuo to yield a tan
solid. The crude product was recrystallized from MeOH (10 mL). The
crystals were filtered and washed with cold MeOH to afford 1.4 g
(60%) of 2-chloro-2-methoxy-benzamide.
[0593] To a solution of the above 2-chloro-2-methoxy-benzamide (1.0
g, 5.4 mmol) in anhydrous THF (10 mL) was added dropwise 1M
BH.sub.3-THF complex (12.8 nm, 12.8 mmol) at 0.degree. C. After 10
min. of stirring at 0.degree. C., the reaction mixture was heated
at 65.degree. C. for 4 h. MeOH (approximately 10 mL) was then added
to the cooled reaction mixture at 0.degree. C. The reaction was
warmed to room temperature and stirred for 10 min. The reaction
solution was concentrated in vacuo to yield an oily product. The
product was dissolved in 50% THF/MeOH (10 mL) and 4M HCl (10 mL)
was added slowly to the solution. After 10 min. the mixture was
partitioned between water (10 mL) and CH.sub.2Cl.sub.2 (20 mL). The
aqueous layer was extracted with CH.sub.2Cl.sub.2 (10 mL) and the
organic layers were discarded. The aqueous layer was basified to pH
9 with 5M NaOH solution. The solution was extracted with
CH.sub.2Cl.sub.2 (3.times.20 mL). The combined organic layers were
washed with brine, dried over MgSO.sub.4, and concentrated in vacuo
to afford 0.36 g (39%) of 5-chloro-2-methoxy-benzylamine as a white
solid. [0594]
(1R,3S)-3-{[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin--
2-yl)amino]methyl}-cyclohexanol, m/z 379.4 [M+1].sup.+
Preparation of cis-3-aminomethyl-cyclohexanol intermediate
[0595] To a solution of 2-cyclohexen-1-one (6 mL, 62 mmol) in
hexanes (100 mL) was added 1M Et.sub.2AlCN (160 mL) in toluene with
cooling at -15.degree. C. After stirring for 30 min, the reaction
mixture was poured into an ice-cold 2M HCl solution and extracted
with CH.sub.2Cl.sub.2. The extracts were washed with cold 2M NaOH
and water, dried and concentrated in vacuo. Distillation of the
product yielded 1.7 g (22%) of 3-oxo-cyclohexanecarbonitrile, b.p.
145-146.degree. C. (16 mmHg).
[0596] To a solution of the above 3-oxo-cyclohexanecarbonitrile
(0.31 g, 2.5 mmol) in MeOH (10 mL) was added NaBH.sub.4 (0.11 g,
2.9 mmol) at 0.degree. C. in one portion. The reaction mixture was
allowed to slowly warm to room temperature and stirred for 2 h.
Excess hydride was consumed by the addition of H.sub.2O. The
reaction mixture was then concentrated under reduced pressure. The
resulting residue was diluted with EtOAc and the solution was
washed with water. The organic phase was washed with brine, dried
over anhydrous Na.sub.2SO.sub.4 and concentrated under reduced
pressure. The crude residue was purified by silica gel
chromatography using a 0-401% gradient of EtOAc and hexanes as an
eluent to provide 0.2 g (64%) of
cis-3-hydrocy-cyclohexanecarbonitrile as a colorless oil.
[0597] To a 2 N solution of NH.sub.3 in MeOH (15 mL) in a high
pressure Parr bottle was added
cis-3-hydroxy-cyclohexanecarbonitrile (0.2 g, 1.6 mmol) and W-7
Raney Nickel (0.35 g, 6.0 mmol). The mixture was placed under 45
psi H.sub.2 and shaken for 15 h. The pressure was released and the
reaction filtered through a pad of diatomaceous earth. The filter
pad was washed with MeOH and the filtrate was concentrated under
reduced pressure to provide 0.16 g (64%) of
cis-3-aminomethyl-cyclohexanol as a white solid [0598]
(1S,3S)-3-{[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin--
2-yl)amino]methyl}-cyclohexanol, m/z 379.5 [M+1].sup.+
Preparation of trans-3-aminomethyl-cyclohexanol intermediate
[0599] To a solution of benzoic acid (1.1 g, 8.8 mmol) and
PS-triphenylphosphine (6.7 g, 15.5 mmol) in anhydrous THF (40 mL)
was added cis-3-hydroxy-cyclohexanecarbonitrile (1.1 g, 8.8 mmol)
in anhydrous THF (20 mL). The reaction mixture was cooled while
diethyl azodicarboxylate (DEAD) (1.9 mL, 12.3 mmol) was added by
syringe over 3 min. The reaction was shaken for 12 h. The mixture
was filtered and the resin was washed with ether. The filtrate was
concentrated in vacuo to afford an oily solid. Silica gel column
chromatography using 95:5 hexanes:EtOAc as an eluent afforded 0.29
g (14%) of benzoic acid trans-3-cyano-cyclohexyl ester as a white
solid.
[0600] A mixture of the above benzoic acid trans-3-cyano-cyclohexyl
ester (0.29 g, 1.3 mmol) and 0.5M NaOCH.sub.3 (0.13 mL, 0.065 mmol)
in MeOH (1 mL) was stirred at room temperature for 12 h. The
reaction mixture was concentrated in vacuo and the resulting
residue was purified by silica gel column chromatography using 3:2
hexanes:EtOAc as an eluent to afford 0.12 g (78%) of
trans-3-hydroxy-cyclohexanecarbonitrile as a colorless oil.
[0601] To a 2 N solution of NH.sub.3 in MeOH (15 mL) in a high
pressure Parr bottle was added
trans-3-hydroxy-cyclohexanecarbonitrile (0.12 g, 1.0 mmol) and W-7
Raney Nickel (0.2 g, 3.4 mmol). The mixture was placed under 45 psi
H.sub.2 and shaken for 15 h. The pressure was released and the
reaction filtered through a pad of diatomaceous earth. The filter
pad was washed with MeOH and the filtrate was concentrated under
reduced pressure to provide 0.1 g (79%) of
trans-3-aminomethyl-cyclohexanol as a white solid. [0602]
2-[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)amin-
o]-1-phenylethanone, m/z 385.3 [M+1].sup.+ [0603]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[5-chloro-2-(trifluorom-
ethoxy)benzyl]-5-nitropyrimidine-2,4-diamine, m/z 475.5
[M+1].sup.+
Preparation of 5-chloro-2-trifluoromethoxy-benzylamine
intermediate
##STR00095##
[0605] Nitronium tetrafluoroborate (14.0 g, 105 mmol) was added to
a solution of 2-trifluoromethoxy-benzaldehyde (10.0 g, 52.6 mmol)
in nitromethane (30 mL) at room temperature. The mixture was
stirred for 2 h and was then quenched with ice water and extracted
with ether (100 mL). The organic layer was separated, washed with
water, dried over MgSO.sub.4 and chromatographed with 5%
EtOAc/hexanes to afford 7.52 g of
5-nitro-2-trifluoromethoxy-benzaldehyde (60.8%) as a pale yellow
solid.
[0606] To a solution of 5-nitro-2-trifluoromethoxy-benzaldehyde
(2.33 g, 9.91 mmol) in MeOH (30 mL) cooled to 0.degree. C. was
added sodium borohydride (440 mg, 11.7 mmol) as a solid in one
portion. The reaction was allowed to slowly warm to room
temperature and stirred for 2 h. Excess hydride was consumed by the
addition of water and the reaction mixture was concentrated under
reduced pressure. The residue was taken back up in EtOAc and washed
with water. The aqueous phase was back extracted with EtOAc and the
combine organic phase was washed with brine, dried over anhydrous
Na.sub.2SO.sub.4 and concentrated under reduced pressure. The crude
residue was purified by flash silica gel chromatography using a
0-40% gradient of EtOAc:hexanes to provide, after concentration of
the eluent, 1.89 g (80%) of
(5-nitro-2-trifluoromethoxy-phenyl)-methanol as a white solid.
[0607] (5-Nitro-2-trifluoromethoxy-phenyl)-methanol (2 g, 8.4 mmol)
was dissolved in EtOAc (50 mL) and was degassed. Pd/C (200 mg) was
then added and the mixture was degassed again. The solution was
hydrogenated with a balloon for 3 h. After the reaction was
complete, the solution was filtered through diatomaceous earth and
the residue was chromatographed with 95:5 CH.sub.2Cl.sub.2:MeOH as
an eluent to give 1.67 g (96%) of
(5-amino-2-trifluoromethoxy-phenyl)-methanol as a yellow solid.
[0608] t-Butyl nitrite (2.49 g, 24.2 mmol), copper (II) chloride
(2.71 g, 20.2 mmol), and acetonitrile (100 mL) were added to a
round-bottom flask. The resulting mixture was cooled to 0.degree.
C. (5-Amino-2-trifluoromethoxy-phenyl)-methanol (1.67 g, 8.1 mmol)
in acetonitrile (10 mL) was slowly added to the reaction over a
period of 5 min. The reaction was warmed up to room temperature and
stirred overnight. The mixture was then poured into 20% HCl (100
mL) and extracted with ether (100 mL). The organic layer was washed
with 20% HCl (100 mL) and dried over MgSO.sub.4. The ether was
removed in vacuo and the residue was chromatographed with 4:1
hexanes:EtOAC to give 1.2 g (66%) of
(5-chloro-2-trifluoromethoxy-phenyl)-methanol as a yellow solid.
DBU (1.0 mL, 6.7 mmol) was added to a solution of
(5-chloro-2-trifluoromethoxy-phenyl)-methanol (1.2 g, 5.3 mmol) and
DPPA (1.4 mL, 6.4 mmol) in toluene (30 mL) at room temperature.
After 3 h, the reaction was concentrated in vacuo to yield an
yellow oil. Silica gel column chromatography using 95:5
hexanes:EtOAc yielded 0.86 g (65%) of
2-azidomethyl-4-chloro-1-trifluoromethoxy-benzene as a colorless
oil.
[0609] Triphenylphosphine (0.99 g, 3.8 mmol) was added to a
solution of the azide (0.86 g, 3.4 mmol) in THF (25 mL) at
0.degree. C. After 5 min, the mixture was warmed to room
temperature. After 18 h, the reaction was diluted with ammonium
hydroxide (10 mL). After 3 h, the reaction was diluted with 3 M
NaOH and stirred for 1 h. The reaction was acidified to pH 2 with 4
N HCl. The solution was then diluted with Et.sub.2O and the layers
were separated. The aqueous layer was extracted with Et.sub.2O and
then basified with 2 N NaOH. The basic aqueous solution was
extracted with CH.sub.2Cl.sub.2 (3.times.50 mL). The combined
organic layers were dried over MgSO.sub.4, filtered and
concentrated to yield an oily solid. The crude solid was diluted
with Et.sub.2O (50 mL) and filtered. The filtrate was concentrated
in vacuo to yield a pale yellow oil. Silica gel column
chromatography using 20:1 CH.sub.2Cl.sub.2:MeOH as an eluent
yielded 0.64 g (83%) of 5-chloro-2-trifluoromethoxy-benzylamine as
a colorless oil. The product was not stable at room temperature and
was used for the next step immediately. [0610]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(4-methoxypyridin-3-yl-
)methyl]-5-nitropyrimidine-2,4-diamine, m/z 388.9 [M+1].sup.+
Preparation of (4-methoxy-pyridin-3-yl)-methylamine
intermediate
##STR00096##
[0612] To a stirred solution of 1.7 M t-butyl lithium in THF (35.0
mL, 59.6 mmol) with THF (150 mL) at -78.degree. C. was added
dropwise 2-bromomesitylene (4.6 mL, 29.8 mmol). After stirring for
1 h, 4-methoxy-pyridine (2.5 g, 22.9 mmol) was added dropwise. This
solution was warmed to -23.degree. C. and stirred for 3 h and then
cooled to -78.degree. C. again. DMF (2.7 mL, 34.4 mmol) was added
and the solution was stirred at -78.degree. C. for 1 h. The
reaction was quenched at this temperature with brine (150 mL) and
extracted with ether. The combined ether was dried over
K.sub.2CO.sub.3 and evaporated in vacuo. The residue was
chromatographed with 50%-80% EtOAc/hexanes to give 2.1 g (65%) of
4-methoxy-pyridine-3-carbaldehyde as a yellow solid.
[0613] To a solution of 4-methoxy-pyridine-3-carbaldehyde (2.1 g,
14.9 mmol) in MeOH (70 mL) cooled to 0.degree. C. was added sodium
borohydride (0.67 g, 17.7 mmol) as a solid in one portion. The
reaction was allowed to slowly warm to room temperature and stirred
for 2 h. Excess hydride was consumed by the addition of water and
the reaction mixture was concentrated under reduced pressure. The
residue was taken back up in EtOAc and washed with water. The
aqueous phase was back extracted with EtOAc and the combined
organic phase was washed with brine, dried over anhydrous
Na.sub.2SO.sub.4 and concentrated under reduced pressure. The crude
residue was purified by flash silica gel chromatography using a
95:5 CH.sub.2Cl.sub.2:MeOH as an eluent to provide, after
concentration, to give 1.1 g (55%) of
(4-methoxy-pyridin-3-yl)-methanol as a white solid. [0614]
(4-Methoxy-pyridin-3-yl)-methanol was then converted to
C-(4-methoxy-pyridin-3-yl)-methylamine through standard procedures
as described in the previous compound. [0615]
(1R,3R)-3-{[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin--
2-yl)amino]methyl}-4,4-dimethylcyclohexanol, m/z 407.6
[M+1].sup.+.
Preparation of (1R,3R)-3-aminomethyl-4,4-dimethyl-cyclohexanol
[0616] To a solution of 4,4'-dimethyl-2-cyclohexene-1-one and
acetic acid in 95% EtOH (70 mL) warmed to 40.degree. C. was added a
solution of potassium cyanide in H.sub.2O (10 mL). The reaction was
stirred at 40.degree. C. for 4 h. The reaction was cooled to room
temperature and filtered through a pad of silica gel. The filter
pad was washed with EtOAc and the mixture was concentrated under
reduced pressure. The residue was taken back up into H.sub.2O and
washed with EtOAc. The combined organic phase was washed with
brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated under
reduced pressure. The crude reaction product was purified by flash
silica gel chromatography using a 0-20% gradient of EtOAc to
hexanes to provide, after concentration of the eluent, 1.03 g of
2,2-dimethyl-5-oxo-cyclohexanecarbonitrile as a clear oil.
[0617] To a solution of 2,2-dimethyl-5-oxo-cyclohexanecarbonitrile
in MeOH (15 mL) cooled to 0.degree. C. was added NaBH.sub.4 as a
solid in one portion. The reaction was allowed to slowly warm to
room temperature and stirred for 2 h. Excess hydride was consumed
by the addition of H.sub.2O and the reaction mixture was
concentrated under reduced pressure. The residue was taken back up
in EtOAc and washed with H.sub.2O. The aqueous phase was back
extracted with EtOAc and the combine organic phase was washed with
brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated under
reduced pressure. The crude residue was purified by flash silica
gel chromatography using a 0-40% gradient of A (EtOAc) to B
(hexanes) to provide, after concentration of the eluent, 400 mg of
(1R,5R)-5-hydroxy-2,2-dimethyl-cyclohexanecarbonitrile as a clear
oil. .sup.1H-NMR indicates only a single diastereomer was
isolated.
[0618] To a 2 N solution of NH.sub.3 in MeOH (15 mL) in a high
pressure Parr bottle was added
(1R,5R)-5-hydroxy-2,2-dimethyl-cyclohexanecarbonitrile and Ni. The
mixture was placed under 45 psi H.sub.2 and shaken for 15 h. The
pressure was released and the reaction filtered through a pad of
diatomaceous earth. The fitter pad was washed with MeOH and the
mixture was concentrated under reduced pressure to provide 260 mg
of a colorless oil. 1H-NMR showed a mixture of
(1R,3R)-3-aminomethyl-4,4-dimethyl-cyclohexanol (major) with a
minor impurity of
(1R,5R)-5-hydroxy-2,2-dimethyl-cyclohexanecarbonitrile. The crude
reaction product was used without further purification. [0619]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(5-bromopyridin-3-yl)m-
ethyl]-5-nitropyrimidine-2,4-diamine, m/z 437.9 [M+1].sup.+.
Preparation of (5-Bromo-pyridin-3-yl)-methylamine
##STR00097##
[0621] To a solution of (5-bromo-pyridin-3-yl)-methanol (2.75 g,
14.6 mmol) and Et.sub.3N (3.10 mL, 22.2 mmol) in DCM (75 mL) at
-20.degree. C. under N.sub.2 was added methanesulfonyl chloride
(1.70 mL, 22.2 mmol) dropwise. After 45 min the reaction was
allowed to warm to room temperature and diluted with DCM (75 mL).
The reaction mixture was washed with water (75 mL), sat.
NaHCO.sub.3 (2.times.75 mL) and brine before drying over
Na.sub.2SO.sub.4. Concentration in vacuo afforded crude
methanesulfonic acid 5-bromo-pyridin-3-ylmethyl ester (4.21 g) as
an oil. The crude material was used in the next step without
purification.
[0622] To a solution of crude methanesulfonic acid
5-bromo-pyridin-3-ylmethyl ester (4.20 g) in DMF (60 mL) was added
NaN.sub.3 (10.0 g, 153.8 mmol). The mixture was stirred under
N.sub.2 overnight then diluted with water. The mixture was
extracted with EtOAc (2.times.300 mL) and the combined organic
layers washed with water before drying over Na.sub.2SO.sub.4. The
solution was concentrated in vacuo to afford crude
3-azidomethyl-5-bromopyridine (2.04 g) as a dark brown oil. This
crude material was used directly in the next step.
[0623] To a solution of crude 3-azidomethyl-5-bromo-pyridine (2.04
g) in THF (50 mL) and water (1 mL) was added triphenylphosphine
(5.02 g, 19.1 mmol). The mixture was heated at reflux under N.sub.2
for 2 h before cooling to room temperature and concentrating in
vacuo. The residue was purified by column chromatography using an
ISCO combi-flash cartridge (silica gel, 100:0 to 30:70 DCM/10%
NH.sub.4OH in MeOH) to afford (5-bromo-pyridin-3-yl)-methylamine
(0.97 g) [0624]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(pyridin-2-ylme-
thyl)pyrimidine-2,4-diamine m/z 358.4 [M+1].sup.+ [0625]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(1,3-thiazol-2--
ylmethyl)pyrimidine-2,4-diamine m/z 364.3 [M+1].sup.+ [0626]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2
(1,3-thiazol-2-ylmethyl)pyrimidine-2,4-diamine m/z 347.6
[M+1].sup.+
[0627] Preparation of (1H-Imidazol-2-yl)-methylamine intermediate
was synthesized as described in literature. (Gebert, U; Von
Kerekjarto, B. Liebigs Ann. Chem. 1968 249-259; Bastiaansen, L. A.
M.; Godefroi, E. F. J. Org. Chem. 1978 1603-1604.) [0628]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(3-methylpyridin-2-yl)-
methyl]-5-nitropyrimidine-2,4-diamine m/z 372.4 [M+1].sup.+ [0629]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(6-methylpyridin-2-yl)-
methyl]-5-nitropyrimidine-2,4-diamine m/z 372.4 [M+1].sup.+ [0630]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(5-methylpyridin-3-yl)-
methyl]-5-nitropyrimidine-2,4-diamine m/z 372.5 [M+1].sup.+
Preparation of (5-Methyl-pyridin-2-yl)-methylamine intermediate
##STR00098##
[0632] To a solution of 5-methyl-pyridine-2-carbonitrile (0.20 g,
1.69 mmol) in THF (10 mL) was added Borane-THF solution (1.0 M
solution in THF, 8.47 mL, 8.47 mmol) rapidly via syringe at room
temperature. The reaction mixture was stirred at room temperature
for 15 min then heated at reflux for 3.5 h. The reaction mixture
was cooled to 0.degree. C., and MeOH (3 mL) was added to the
reaction mixture slowly. After the r.times.n mixture was stirred
for 10 min, the mixture was then concentrated to a white solid
residue using reduced pressure. The residue was then suspended in 5
mL of 1:1 THF/MeOH and treated with 2 mL of 4 M HCl, stirred at
room temperature for 10 min, then neutralized to basic pH by the
addition of 2.5 M NaOH. The mixture was extracted with
CH.sub.2Cl.sub.2. the combined organic layers were washed with
brine, dried over MgSO.sub.4, filtered and concentrated to afford
0.10 g of (5-methyl-pyridin-2-yl)-methylamine as a colorless oil.
The crude reaction product was used without further
purification.
[0633] The following two intermediates were prepared using similar
procedures as described above: [0634]
(6-Methyl-pyridin-3-yl)-methylamine [0635]
(5-Methyl-pyridin-3-yl)-methylamine [0636]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(5-chloropyridin-2-yl)-
methyl]-5-nitropyrimidine-2,4-diamine m/z 372.5 [M+1].sup.+ [0637]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(6-methylpyridin-3-yl)-
methyl]-5-nitropyrimidine-2,4-diamine m/z 372.6 [M+1].sup.+ [0638]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(5-methyl-1,3-oxazol-4-
-yl)methyl]-5-nitropyrimidine-2,4-diamine m/z 362.3 [M+1].sup.+
[0639] (5-Methyl-oxazol-4-yl)-methylamine was synthesized from the
(5-methyl-oxazol-4-yl)-methanol using the method described in the
synthesis of 5-chloro-2-trifluoromethoxy-benzylamine
##STR00099##
Example 4
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(2-phenylethyl)p-
yrimidine-2,4-diamine
##STR00100##
[0641] To a solution of 2-chloro-5-nitro-4-thiocyanato-pyrimidine
(54 mg, 0.25 mmol) in CH.sub.2Cl.sub.2 were added 2-phenethylamine
(30 mg, 0.25 mmol) and DIPEA (130 ml, 0.75 mmol). The reaction
mixture was stirred at room temperature for 16 h until the reaction
was complete. The reaction mixture was then used for the next
reaction without purification.
[0642] To a solution of the above
(5-nitro-4-thiocyanato-pyrimidin-2-yl)-phenethyl-amine (75 mg, 0.25
mmol) in CH.sub.2Cl.sub.2 was added tert-butyl
trans-4-aminomethylcyclohexylcarbamate (114 mg, 0.5 mmol). The
reaction mixture was stirred at room temperature for 16 h and
concentrated in vacuo. The resulting residue was then diluted with
CH.sub.2Cl.sub.2 (1 mL) and TFA (0.5 mL) was added. The mixture was
stirred at room temperature for 4 h and concentrated in vacuo. The
resulting residue was purified by preparative LCMS to afford 36 mg
(39%) of the title compound, m/z 371.6 [M+1].sup.+.
[0643] The following compound was prepared using similar procedures
as described above: [0644]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(3-chlorophenyl)ethy-
l]-5-nitropyrimidine-2,4-diamine, m/z 405.6 [M+1].sup.+ [0645]
4-{2-[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)a-
mino]ethyl}phenol, m/z 387.4 [M+1].sup.+
Example 5
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(trifluoromethoxy)ben-
zyl]-5-(trifluoromethyl)pyrimidine-2,4-diamine
##STR00101##
[0647] To a solution of 5-trifluoromethyl)-2,4-di-chloropyrimidine
(0.5 g, 2.3 mmol) in DMF (8 mL) were added a solution of
2-(trifluoromethoxy)benzylamine (0.44 g, 2.3 mmol) in DMF (2 mL)
followed by DIPEA (0.44 mL, 2.3 mmol) at -20.degree. C. The
reaction mixture was stirred at that temperature for 20 min. When
the starting material was all consumed, the mixture was diluted
with EtOAc and washed with water (.times.4). The organic phase was
dried over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo.
The resulting residue was purified by silica gel preparative TLC
using 100% CH.sub.2Cl.sub.2 as an eluent to afford 0.44 g (52%) of
(4-chloro-5-trifluoromethyl-pyrimidin-2-yl)-(2-trifluoromethoxy-benzyl)-a-
mine as a white foam.
[0648] To a solution of the above
(4-chloro-5-trifluoromethyl-pyrimidin-2-yl)-(2-trifluoromethoxy-benzyl)-a-
mine (80 mg, 0.22 mmol) in DMF (2 mL) was added the Boc-protected
cyclohexyl amine (74 mg, 0.33 mmol) followed by DIPEA (37 .mu.L,
0.22 mmol). The reaction mixture was stirred at 70.degree. C. for 3
h. When the reaction was complete, the reaction mixture was diluted
with EtOAc and washed with water (.times.4). The organic phase was
dried over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo.
The resulting residue was purified by silica gel preparative TLC
using 95:5 CH.sub.2Cl.sub.2:MeOH as an eluent. The resulting
residue was then re-dissolved in CH.sub.2Cl.sub.2 (10 mL) and TFA
(1 mL) was added. The reaction mixture was stirred at room
temperature for 2 h and treated with saturated NaHCO.sub.3. The
product was extracted with CH.sub.2Cl.sub.2 and dried over
anhydrous Na.sub.2SO.sub.4. Silica gel preparative TLC using
CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH 10:1:0.2 as an eluent afforded 56
mg (56%) of the title compound as a white solid, m/z 462.4
[M-1].sup.+.
Example 6
N.sup.4-[(trans-4-pyrrolidin-1-ylcyclohexyl)methyl]-N.sup.2-[2-(trifluorom-
ethoxy)benzyl]-5-(trifluoromethyl)pyrimidine-2,4-diamine
##STR00102##
[0650] To a solution of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(trifluoromethoxy)be-
nzyl]-5-(trifluoromethyl)pyrimidine-2,4-diamine (100 mg, 0.22 mmol)
in DMF (2 mL) were added 1,4-dibromobutane (140 mg, 0.65 mmol) and
DIPEA (113 .mu.L, 0.65 mmol). The reaction mixture was stirred at
room temperature for 16 h and then heated at 65.degree. C. for
another 2 h. The reaction mixture was then diluted with EtOAc and
washed with water (.times.4). The organic phase was dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The resulting
residue was purified by silica gel preparative TLC using 10:1
CH.sub.2Cl.sub.2:MeOH as an eluent to afford 49 mg of the title
compound as a while solid, m/z 518.6 [M+1].sup.+.
[0651] The following compounds were prepared using similar
procedures as described above: [0652]
2,2'-({trans-4-[({2-[(3-bromo-2-methylbenzyl)amino]-5-nitropyrimidin-4-yl-
}amino)methyl]cyclohexyl}imino)diacetamide, m/z 565.0 [M+1].sup.+
[0653]
N.sup.2-{trans-4-[({2-[(3-bromo-2-methylbenzyl)amino]-5-nitropyrimidin-4--
yl}amino)methyl]cyclohexyl}glycinamide, m/z 508.9 [M+2].sup.+
[0654]
N.sup.2-(3-bromo-2-methylbenzyl)-5-nitro-N.sup.4-{(trans-4-[(2,2,2-triflu-
oroethyl)amino]cyclohexyl}methyl)pyrimidine-2,4-diamine, m/z 532.9
[M+1].sup.+ [0655]
N.sup.2-(3-bromo-2-methylbenzyl)-N.sup.4-({trans-4-[(2,2-difluoroethyl)am-
ino]cyclohexyl}-methyl)-5-nitropyrimidine-2,4-diamine, m/z 514.9
[M+1].sup.+ [0656]
N.sup.4-({trans-4-[(2-fluoroethyl)amino]cyclohexyl}methyl)-5-nitro-N.sup.-
2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 487.0
(M+1).sup.+. [0657]
N.sup.4-({trans-4-[(2,2-difluoroethyl)amino]cyclohexyl}methyl)-5-nitro-N.-
sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 505.0
(M+1).sup.+. [0658]
5-Nitro-N.sup.4-({trans-4-[(2-pyridin-3-ylethyl)amino]cyclohexyl}methyl)--
N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z
546.6 (M+1).sup.+.
Preparation of 3-(2-bromo-ethyl)-pyridine
[0659] A solution of 2-(3-pyridyl)ethan-1-ol (100 mg, 0.79 mmol)
and CBr.sub.4 (314 mg, 0.95 mmol) in THF (2 mL) was cooled to
0.degree. C. and PPh.sub.3 (313 mg, 1.18 mmol) was added
portionwise. The ice-bath was removed and the reaction mixture was
stirred at room temperature for 16 h. The mixture was concentrated
and diluted with CH.sub.2Cl.sub.2. The mixture was basified using
saturated NaHCO.sub.3 and the organic phase aws separated. The
aqueous phase was re-extracted with CH.sub.2Cl.sub.2 and the
combined organic phase was dried over Na.sub.2SO.sub.4 and
concentrated. Silica gel prep TLC using 95:5 CH.sub.2Cl.sub.2:MeOH
as an eluent afforded 3-(2-bromoethyl)-pyridine.
Example 7
5-nitro-N.sup.4-[(trans-4-pyrrolidin-1-ylcyclohexyl)methyl]-N.sup.2-[2-(tr-
ifluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00103##
[0661] To a solution of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]-pyrimidine-2,4-diamine (86 mg, 0.2 mmol) in DMF (1
mL) were added 1,4-dibromobutane (710 .mu.L, 0.59 mmol) and DIPEA
(103 .mu.L, 0.59 mmol). The reaction mixture was stirred at
65.degree. C. for 4 h. The reaction mixture was then diluted with
EtOAc and washed with saturated NaHCO.sub.3 and water (.times.4).
The organic phase was dried over anhydrous Na.sub.2SO.sub.4 and
concentrated in vacuo. The resulting residue was purified by silica
gel preparative TLC using 10:1:0.05
CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an eluent to afford 60 mg (62%)
of the title compound as an off-white solid, m/z 493.3
[M-1].sup.+.
[0662] The following compounds were prepared following similar
procedures as described above: [0663]
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)pyrrolidin-3-ol, m/z 509.3 [M-1].sup.+
[0664]
N.sup.4-[(trans-4-morpholin-4-ylcyclohexyl)methyl]-5-nitro-N.sup.2-[2-(tr-
ifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 511.7 [M+1].sup.+
[0665]
5-nitro-N.sup.4-[(trans-4-piperidin-1-ylcyclohexyl)methyl]-N.sup.2-[(trif-
luoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 509.7 [M+1].sup.+
[0666]
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-5-nitro-N.sup.2-[2-(tri-
fluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 479.5 [M-1].sup.+
[0667]
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)azetidin-3-ol, m/z 497.6 [M+1].sup.+
[0668]
1-(trans-4-{[(2-{([2-chloro-3-(dimethylamino)benzyl]amino}-5-nitropyrimid-
in-4-yl)amino]methyl}cyclohexyl)azetidin-3-ol, m/z 490.6
[M+1].sup.+. [0669]
1-(trans-4-{[(2-{[(4-chloropyridin-3-yl)methyl]amino}-5-nitropyrim-
idin-4-yl)amino]methyl}cyclohexyl)azetidin-3-ol, m/z 448.5
[M+1].sup.+ [0670]
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-N.sup.2-[(2-meth-
oxypyridin-3-yl)methyl]-5-nitropyrimidine-2,4-diamine m/z 428.6
[M+1].sup.+ [0671]
1-(trans-4-{[(2-{[(2-methoxypyridin-3-yl)methyl]amino}-5-nitropyrimidin-4-
-yl)amino]methyl}cyclohexyl)azetidin-3-ol, m/z 444.5 [M+1].sup.+
[0672]
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-N.sup.2-[(4-chloropyrid-
in-3-yl)methyl]-5-nitropyrimidine-2,4-diamine m/z 432.5 [M+1].sup.+
[0673]
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-5-nitro-N.sup.2-{[4-(tr-
ifluoromethyl)pyridin-3-yl]methyl}pyrimidine-2,4-diamine m/z 466.5
[M+1].sup.+ [0674]
1-[trans-4-({[5-nitro-2-({[4-(trifluoromethyl)pyridin-3-yl]methyl}amino)p-
yrimidin-4-yl]amino}methyl)cyclohexyl]azetidin-3-ol m/z 482.5
[M+1].sup.+
Example 8
N.sup.4-{[trans-4-(dimethylamino)cyclohexyl]methyl}-5-nitro-N.sup.2-[2-(tr-
ifluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00104##
[0676] To a solution of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]-pyrimidine-2,4-diamine (100 mg, 0.23 mmol) in
dichloroethane (2 mL) were added 37% formaldehyde (74 mg, 0.91
mmol) and sodium triacetoxyborohydride (144 mg, 0.68 mmol). The
reaction mixture was stirred at room temperature for 2 h. The
reaction mixture was treated with a mixture of saturated
NaHCO.sub.3 and 1M Na.sub.2CO.sub.3 until pH 9 and extracted with
CH.sub.2Cl.sub.2 (.times.2). The combined organic phase was dried
over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The
resulting residue was purified by silica gel preparative TLC using
10:1:0.2 CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an eluent to afford 59
mg of the title compound as a yellowish solid, m/z 469.7
[M+1].sup.+.
[0677] The following compound was prepared following similar
procedures as described above: [0678]
N.sup.4-{[trans-4-(diethylamino)cyclohexyl]methyl}-5-nitro-N.sup.2-[2-(tr-
ifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 497.8 [M+1].sup.+
[0679]
N.sup.4-{[trans-4-(ethylamino)cyclohexyl]methyl}-5-nitro-N.sup.2-[2-(trif-
luoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 469.4
[M+1].sup.+
Example 9
N.sup.4-{[trans-4-(benzylamino)cyclohexyl]methyl}-5-nitro-N.sup.2-[2-(trif-
luoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00105##
[0681] To a solution of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]-pyrimidine-2,4-diamine (100 mg, 0.23 mmol) in DMF (3
mL) were added benzyl bromide (39 mg, 0.23 mmol) followed by DIPEA
(40 .mu.L, 0.23 mmol). The reaction mixture was stirred at room
temperature for 20 h. The reaction mixture was diluted with EtOAc
and washed with water (.times.4). The organic phase was dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The resulting
residue was purified by silica gel prep TLC using 10:1
CH.sub.2Cl.sub.2:MeOH as an eluent to afford 8 mg of the title
compound, m/z 531.6 [M+1].sup.+.
[0682] The following compounds were prepared following similar
procedures as described above. [0683]
5-nitro-N.sup.4-({trans-4-[(pyridin-4-ylmethyl)amino]cyclohexyl}methyl)-N-
.sup.2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine, m/z
532.6 [M+1].sup.+ [0684]
5-nitro-N.sup.4-({trans-4-[(pyridin-3-ylmethyl)amino]cyclohexyl}methyl)-N-
.sup.2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine, m/z
532.4 [M+1].sup.+ [0685]
2-[(trans-4-{[(5-nitro-2-{[2-trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)amino]ethanol, m/z 485.7 [M+1].sup.+
[0686]
2,2'-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin--
4-yl)amino]methyl}-cyclohexyl)imino]diethanol, m/z 529.7
[M+1].sup.+ [0687]
N.sup.4-{[trans-4-(dibenzylamino)cyclohexyl]methyl}-5-nitro-N.sup.-
2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 621.7
[M+1].sup.+ [0688]
N.sup.4-({trans-4-[bis(pyridin-2-ylmethyl)amino]cyclohexyl}methyl)-5-nitr-
o-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z
623.7 [M+1].sup.+ [0689]
N.sup.2-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}-cyclohexyl)glycinamide, m/z 496.3 [M-1].sup.+
[0690]
N.sup.2-[trans-4-({[5-nitro-2-({[4-(trifluoromethyl)pyridin-3-yl]methyl}a-
mino)pyrimidin-4-yl]amino}methyl)cyclohexyl]glycinamide, m/z 483.4
[M+1].sup.+
Example 10
4-{[(trans-4-aminocyclohexyl)methyl]amino}-2-{[2-(trifluoromethoxy)-benzyl-
]amino}pyrimidine-5-carbonitrile
##STR00106## ##STR00107##
[0692] To a mixture of N-cyanoacetylurethane (100 g, 640.5 mmol) in
acetic anhydride (160 mL) was added triethyl orthoformate (26.6 mL,
160 mmol). The mixture was stirred at reflux for 2.5 h; it was then
cooled to 25.degree. C. whereupon crystals formed. The reaction
mixture was filtered and the collected solid was washed with
hexanes and dried under vacuum to afford 21.4 g (67%) of the
desired product.
[0693] Ammonium hydroxide (150 mL) was added to the above
acrylamide (54.5 g, 258.29 mmol) in H.sub.2O (150 mL) at 25.degree.
C. followed by NaOH (32 g) in H.sub.2O (150 mL) dropwise over 10
min. The mixture was then heated at 70.degree. C. with stirring for
1 h and then cooled to 25.degree. C. The reaction mixture was
filtered while basic, and then acidified to pH 4 with HCl and
filtered to yield 25.4 g of uracil.
[0694] To a mixture of the above uracil (15.41 g, 112.5 mmol) and
phosphorus oxychloride (157.4 mL, 1.69 mol) was added
N,N-diethylaniline (34.2 mL, 225 mmol), cautiously, at 25.degree.
C. The reaction was then heated at 115.degree. C. for 3 h after
which the mixture was cooled to 40.degree. C. and concentrated in
vacuo to remove excess POCl.sub.3. The remaining liquid was poured
slowly onto ice-water with rapid stirring and the precipitated
product was filtered. The aqueous layer was then extracted with
chloroform and the extract was washed with 1 N HCl before
concentrating in vacuo. This material was combined with the
previously filtered material to afford
2,4-dichloro-pyrimidine-5-carbonitrile (15.36 g, 79%).
[0695] 2,4-Dichloro-pyrimidine-5-carbonitrile (50 mg, 0.287 mmol)
was dissolved in DMF (1 mL) and 2-(trifluoromethoxyl)benzylamine
(57 mg, 0.287 mmol) was added to the solution followed by DIPEA (51
.mu.L, 0.287 mmol). The reaction mixture was stirred at room
temperature for 1 h. The solution was then diluted with EtOAc and
washed with water (.times.4). The organic phase was then dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was purified by silica gel prep TLC using CH.sub.2Cl.sub.2 as an
eluent to afford 51 mg (54%) of
(4-chloro-5-nitrile-pyrimidin-2-yl)-(2-trifluoromeoxy-benzyl)-amine
as a white solid.
[0696]
(4-Chloro-5-cyanopyrimidin-2-yl)-(2-trifluoromethoxy-benzyl)-amine
(50 mg, 0.16 mmol) was dissolved in DMF (1 mL) and tert-butyl
trans-4-aminomethylcyclohexylcarbamate (70 mg, 0.30 mmol) was added
followed by DIPEA (53 .mu.L, 0.30 mmol). The reaction mixture was
stirred at room temperature for 16 h. The reaction mixture was then
diluted with EtOAc and washed with water (.times.3). The organic
phase was dried over anhydrous Na.sub.2SO.sub.4 and concentrated in
vacuo. The resulting residue was purified by silica gel preparative
TLC using 95:5 CH.sub.2Cl.sub.2:MeOH as an eluent to afford 76 mg
(96%) of
(4-{[5-cyano-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylamino]-meth-
yl}-cyclohexyl)-carbamic acid tert-butyl ester.
[0697] The above tert-butyl ester (75 mg, 0.14 mmol) was dissolved
in CH.sub.2Cl.sub.2 (10 mL) and TFA (0.7 mL) was added to the
solution. The reaction mixture was stirred at room temperature for
1 h and quenched with saturated NaHCO.sub.3. The organic phase was
separated from the aqueous phase which was then re-extracted with
CH.sub.2Cl.sub.2 (.times.2). The combined organic phase was dried
over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The
resulting residue was purified by silica gel preparative TLC using
10:1:0.1 CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an eluent to afford 46
mg (77%) of the title compound, m/z 419.4 (M-1).sup.+.
[0698] The following compounds were prepared using similar
procedures as described above: [0699]
4-{[(trans-4-aminocyclohexyl)methyl]amino}-2-{[(2-chloropyridin-3-yl)meth-
yl]amino}pyrimidine-5-carbonitrile, m/z 372.8 [M+1[.sup.+ [0700]
4-{[(trans-4-aminocyclohexyl)methyl]amino}-2-{[(2-methoxypyridin-3-yl)met-
hyl]amino}pyrimidine-5-carbonitrile, m/z 368.7 [M+1].sup.+ [0701]
4-{[(trans-4-aminocyclohexyl)methyl]amino}-2-{[(4-methoxypyridin-3-yl)met-
hyl]amino}pyrimidine-5-carbonitrile m/z 368.6 [M+1].sup.+
Example 11
4-({[trans-4-(dimethylamino)cyclohexyl]methyl}amino)-2-{[2-(trifluorometho-
xy)benzyl]amino}pyrimidine-5-carbonitrile
##STR00108##
[0703] To a solution of
4-{[(trans-4-aminocyclohexyl)methyl]amino}-2-{[2-(trifluoromethoxy)benzyl-
]-amino}pyrimidine-5-carbonitrile (100 mg, 0.24 mmol) in
dichloroethane (2 mL) were added 37% formaldehyde (100 mg, 1.2
mmol) and NaBH(OAc).sub.3 (151 mg, 0.71 mmol). The reaction mixture
was stirred at room temperature for 2 h. The reaction mixture was
then basified with a mixture of saturated NaHCO.sub.3 and 1M
Na.sub.2CO.sub.3 to pH 9. The organic phase was washed with water
(.times.1), dried over anhydrous Na.sub.2SO.sub.4 and concentrated
in vacuo. The resulting residue was purified by silica gel
preparative TLC using 10:1:0.2 CH.sub.2Cl.sub.2;MeOH:NH.sub.4OH as
an eluent to afford 39 mg (36%) of the title compound as a while
solid, m/z 449.5 (M+1).sup.+.
Example 12
4-({[trans-4-(ethylamino)cyclohexyl]methyl}amino)-2-{[2-(trifluoromethoxy)-
benzyl]amino}pyrimidine-5-carbonitrile
##STR00109##
[0705]
4-{[(trans-4-Aminocyclohexyl)methyl]amino}-2-{[2-(trifluoromethoxy)-
benzyl]amino}pyrimidine-5-carbonitrile (150 mg, 0.36 mmol) was
dissolved in a solution of 1% AcOH in MeOH. The solution was cooled
to 0.degree. C. and to this cold solution were added
NaBH(OAc).sub.3 and acetaldehyde. The reaction mixture was stirred
at room temperature for 16 h. The reaction mixture was then treated
with saturated NaHCO.sub.3 and extracted with EtOAc. The organic
phase was dried over anhydrous Na.sub.2SO.sub.4 and concentrated in
vacuo. The resulting residue was purified by silica gel column
chromatography using 9:1:0.1 CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an
eluent to afford 24 mg (15%) of the title compound, m/z 449.4
(M+1).sup.+.
[0706] The following compounds were prepared following similar
procedures as described above. [0707]
4-({[trans-4-(diethylamino)cyclohexyl]methyl}amino)-2-{[2-(trifluorometho-
xy)benzyl]amino}-pyrimidine-5-carbonitrile m/z 477.7 (M+1).sup.+.
[0708]
4-[({trans-4-[(pyridin-2-ylmethyl)amino]cyclohexyl}methyl)amino]-2-{[2-(t-
rifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile, m/z 510.7
[M-1].sup.+. [0709]
4-[({trans-4-[(pyridin-3-ylmethyl)amino]cyclohexyl}methyl)amino]-2-{[2-(t-
rifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile, m/z 510.5
[M-1].sup.+. [0710]
4-[({trans-4-[(pyridin-4-ylmethyl)amino]cyclohexyl}methyl)amino]-2-{[2-(t-
rifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile, m/z 510.4
[M-1].sup.+. [0711]
4-[({trans-4-[(2,2-dimethylpropyl)amino]cyclohexyl}methyl)amino]-2-{[2-(t-
rifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile, m/z 489.4
[M-1].sup.+. [0712]
4-[({trans-4-[(cyclopropylmethyl)amino]cyclohexyl)methyl}amino]-2-{[2-(tr-
ifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile, m/z 475.7
[M-1].sup.+. [0713]
4-({[trans-4-(isopropylamino)cyclohexyl]methyl}amino)-2-{[2-(trifluoromet-
hoxy)benzyl]-amino}pyrimidine-5-carbonitrile, m/z 461.5
[M-1].sup.+.
Example 13
4-{[(trans-4-azetidin-1-ylcyclohexyl)methyl]amino}-2-{[2-(trifluoromethoxy-
)benzyl]amino}pyrimidine-5-carbonitrile
##STR00110##
[0715] To a solution of
4-{[(trans-4-aminocyclohexyl)methyl]amino}-2-{[2-(trifluoromethoxy)benzyl-
]-amino}pyrimidine-5-carbonitrile (150 mg, 0.36 mmol) in DMF (1 mL)
were added 1,3-dibromo-propane (86 mg, 0.43 mmol) and DIPEA (124
.mu.L, 0.71 mmol). The reaction mixture was stirred at 50.degree.
C. for 24 h. The reaction mixture was then diluted with EtOAc and
washed with NaHCO.sub.3 (.times.1) and water (.times.4). The
organic phase was dried over anhydrous Na.sub.2SO.sub.4 and
concentrated in vacuo. The resulting residue was purified by silica
gel preparative TLC using 10:1:0.2 CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH
as an eluent to afford 54 mg (33%) of the title compound as a white
solid, m/z 461.6 (M+1).sup.+.
[0716] The following compounds were prepared using similar
procedures described above: [0717]
4-{[(trans-4-pyrrolidin-1-ylcyclohexyl)methyl]amino}-2-{[2-(trifluorometh-
oxy)benzyl]amino}-pyrimidine-5-carbonitrile, m/z 473.4 (M-1).sup.+
[0718]
4-({[trans-4-(3-hydroxypyrrolidin-1-yl)cyclohexyl]methyl}amino)-2-{[2-(tr-
ifluoromethoxy)-benzyl]amino}pyrimidine-5-carbonitrile, m/z 489.2
(M-1).sup.+ [0719]
4-({[trans-4-(3-hydroxyazetidin-1-yl)cyclohexyl]methyl}amino)-2-{[2-(trif-
luoromethoxy)benzyl]-amino}pyrimidine-5-carbonitrile, m/z 475.4
[M-1].sup.+
Example 14
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-{2-[(2-aminophenyl)thio]-
benzyl}-5-nitropyrimidine-2,4-diamine
##STR00111##
[0721] To a solution of
{4-[(2-chloro-5-nitro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}-carbamic
acid tert-butyl ester (100 mg, 0.26 mmol) in a mixture of
CH.sub.2Cl.sub.2 (2 mL) and DMF (1 mL) were added 2-iodobenzylamine
(192 mg, 0.82 mmol) and DIPEA (135 .mu.L, 1.04 mmoL). The reaction
mixture was stirred at room temperature for 16 h and concentrated
in vacuo. The resulting residue was diluted with EtOAc and washed
with water (.times.3) and then with brine. The organic phase was
dried over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo.
The residue was purified by silica gel preparative TLC using 98:2
CH.sub.2Cl.sub.2:MeOH as an eluent to afford 97 mg (64%) of
(4-{[2-(2-iodo-benzylamino)-5-nitro-pyrimidin-4-ylamino]-methyl}-cyclohex-
yl)-carbamic acid tert-butyl ester.
[0722] A mixture of the above
(4-{[2-(2-iodo-benzylamino)-5-nitro-pyrimidin-4-ylamino]-methyl}-cyclohex-
yl)-carbamic acid tert-butyl ester (95 mg, 0.16 mmol), CuI (3 mg,
0.016 mmol), K.sub.2CO.sub.3 (50 mg, 0.36 mmol), ethylene glycol
(27 .mu.L, 0.49 mmol) in isopropanol (4 mL) was placed in a sealed
tube and heated in a microwave at 150.degree. C. for 2 h. The
reaction mixture was treated with water (6 mL) and the organic
layer was separated. The aqueous layer was extracted with EtOAc
(.times.3). The combined organic layer was dried over anhydrous
Na.sub.2SO.sub.4 and concentrated in vacuo. The resulting residue
was purified by silica gel preparative TLC using 95:5
CH.sub.2Cl.sub.2:MeOH as an eluent to afford 35 mg (37%) of
[4-({2-[2-(2-amino-phenylsulfanyl)-benzylamino]-5-nitro-pyrimidin-4-ylami-
no}-methyl)-cyclohexyl]-carbamic acid tert-butyl ester.
[0723] To a solution of the above
[4-({2-[2-(2-amino-phenylsulfanyl)-benzylamino]-5-nitro-pyrimidin-4-ylami-
no}-methyl)-cyclohexyl]-carbamic acid tert-butyl ester (35 mg, 0.06
mmol) in CH.sub.2Cl.sub.2 (10 mL) was added TFA (1.2 mL). The
reaction mixture was stirred at room temperature for 2 h. The
reaction mixture was then treated with saturated NaHCO.sub.3.
During the work-up, some MeOH was added to solubilize the product
in the organic phase. The organic phase was then dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The resulting
residue was purified by silica gel preparative TLC using 10:1:0.1
CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an eluent to afford 20 mg 70%)
of the title compound as an off-white solid, m/z 477.9
(M-1).sup.+.
Preparation of 2-iodobenzylamine
[0724] To a solution of 2-iodobenzyl alcohol (6.0 g, 25.4 mmol) and
DPPA (6.8 mL, 30.5 mmol) in toluene (65 mL) was added DBU (5.0 mL,
33.0 mmol). The reaction mixture was stirred at room temperature
for 16 h. The reaction mixture was then diluted with 3M HCl (60 mL)
and ether (150 mL). The organic phase was separated and the aqueous
phase was extracted with ether (.times.3). The combined organic
extracts were washed with water (.times.2) and then with brine. The
solution was dried over anhydrous Na2SO4 and concentrated in vacuo.
Silica gel column chromatography using 95:5 hexanes:EtOAc afforded
6.1 g (93%) of 1-azidomethyl-2-iodo-benzene.
[0725] To a solution of 1-azidomethyl-2-iodo-benzene (6.1 g, 23.4
mmol) in anhydrous THF (100 mL) was added triphenylphosphine (6.8
g, 25.7 mmol) at 0.degree. C. The reaction mixture was slowly
warned up to room temperature and stirred for 16 h. The mixture was
diluted with ammonium hydroxide (20 mL) and stirred for 3 h. The
mixture was then treated with 2M NaOH (30 mL) and stirred for 1 h.
The reaction mixture was acidified to pH 2 by adding 3 M HCl. The
mixture was diluted with ether and the layers were separated. The
aqueous layer was basified to pH 9 by adding 2 M NaOH and then
extracted with CH.sub.2Cl.sub.2 (.times.3). The combined organic
phase was dried over anhydrous Na.sub.2SO.sub.4 and concentrated in
vacuo to yield an oily residue. The resulting residue was treated
with ether and filtered. The filtrate was purified by silica gel
column chromatography using 95:5 CH.sub.2Cl.sub.2:MeOH as an eluent
to afford 5.4 g (80%) of 2-iodobenzylamine.
Example 15
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-chloro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]pyrimidine-2,4-diamine
##STR00112##
[0727] To a suspension of 5-chlorouracil (15.0 g, 102.4 mmol) in
POCl.sub.3 (50 mL, 326.1 mmol) was added N,N-diethylaniline (7.5
mL). The reaction mixture was heated at 110.degree. C. for 24 h.
The reaction mixture was cooled to room temperature and
concentrated in vacuo to about 25 mL. The resulting residue was
then poured into ice and stirred until all the ice melted. The
aqueous layer was extracted with ether (.times.3). The combined
organic phase was dried over anhydrous Na.sub.2SO.sub.4 and
concentrated in vacuo. The resulting residue was distilled under
vacuum at .about.90.degree. C. to afford 12.5 g (81%) of
5-chloro-2,4-dichloropyrimidine.
[0728] To a solution of the above 5-chloro-2,4-dichloropyrimidine
(73 mg, 0.4 mmol) in EtOH (2 mL) were added tert-butyl
trans-4-aminomethylcyclohexylcarbamate (100 mg, 0.44 mmol) and
DIPEA (77 .mu.L, 0.44 mmol). The reaction mixture was heated at
40.degree. C. for 1 h. The reaction mixture was concentrated in
vacuo and the resulting residue was diluted with EtOAc. The
solution was washed with brine. The aqueous layer was re-extracted
with EtOAc (.times.2). The combined organic phase was dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo to yield 148
mg (99%) of
{4-[(2,5-dichloro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}-carbamic
acid tert-butyl ester.
[0729] The above
{4-[(2,5-dichloro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}-carbamic
acid tert-butyl ester (97 mg, 0.26 mmol) was dissolved in
2-trifluoromethoxy-benzylamine (2.5 mL) and the mixture was heated
to 180.degree. C. for 20 min. The reaction mixture was diluted with
EtOAc and poured into 1N HCl solution. The aqueous layer was
separated and extracted with EtOAc (.times.2). The combined organic
phase was dried over anhydrous Na.sub.2SO.sub.4 and concentrated in
vacuo. The resulting residue was dissolved in CH.sub.2Cl.sub.2 (2.5
mL) and TFA (2.5 mL) was added. The reaction mixture was stirred at
room temperature for 1 h. The mixture was then poured into 10%
NaHCO.sub.3 and the aqueous layer was extracted with EtOAc
(.times.2). The combined organic phase was dried over anhydrous
Na.sub.2SO.sub.4 and concentrated in vacuo. The resulting residue
was purified by silica gel preparative TLC using 9:1:0.1
CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an eluent to afford 97 mg (87%)
of the title compound, m/z 430.5 (M+1).sup.+.
[0730] The following compounds were prepared using similar
procedures as described above: [0731]
N.sup.4-[(cis-4-aminocyclohexyl)methyl]-5-chloro-N.sup.2-[2-(trifluoromet-
hoxy)benzyl]pyrimidine-2,4-diamine, m/z 430.2 (M+1).sup.+. [0732]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-benzyl-5-chloropyrimidi-
ne-2,4-diamine, m/z 346.5 (M+1).sup.+.
Example 16
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-fluoro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]pyrimidine-2,4-diamine
##STR00113##
[0734] Phosphorous pentachloride (8.0 g, 38.4 mmol) was added to a
mixture of 5-fluorouracil (10.0 g, 76.9 mmol) and phosphorous
oxychloride (30 mL). The reaction mixture was heated at reflux, for
14 h, under N.sub.2. The reaction mixture was cooled to room
temperature and then poured into ice. The remaining residue from
the flask was dissolved in saturated Na.sub.2CO.sub.3 and poured
into the ice mixture. The mixture was extracted with EtOAc
(.times.3) and the organic phase was dried over anhydrous
Na.sub.2SO.sub.4. The solution was concentrated in vacuo to yield
9.1 g (71%) of 5-fluoro-2,4-dichloropyridine as a pale yellow
oil.
[0735] To a solution of the above 5-fluoro-2,4-dichloropyridine (67
mg, 0.4 mol) in EtOH (2 mL) were added tert-butyl
trans-4-aminomethylcyclohexylcarbamate (100 mg, 0.44 mmol) and
DIPEA (77 .mu.L, 0.44 mmol). The reaction mixture was heated at
40.degree. C. for 1 h. The mixture was concentrated in vacuo and
the resulting residue was dissolved in EtOAc and washed with brine.
The aqueous layer was re-extracted with EtOAc (.times.2). The
combined organic phase was dried over anhydrous Na.sub.2SO.sub.4
and concentrated in vacuo to yield 127 mg (89%) of
{4-[(2-chloro-5-fluoro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}--
carbamic acid tert-butyl ester.
[0736]
{4-[(2-Chloro-5-fluoro-pyrimidin-4-ylamino-methyl]-cyclohexyl}-carb-
amic acid tert-butyl ester (127 mg, 0.35 mmol) was dissolved in
2-trifluoromethoxybenzylamine (2.5 mL) and heated at 180.degree. C.
for 20 min. The reaction mixture was diluted with EtOAc and poured
into 1N HCl. The aqueous layer was separated and extracted with
EtOAc (.times.2). The combined organic phase was dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The resulting
residue was dissolved in CH.sub.2Cl.sub.2 (2.5 mL) and TFA (2.5 mL)
was added. The reaction mixture was stirred at room temperature for
1 h. The reaction mixture was concentrated in vacuo and the
resulting residue was diluted with EtOAc. The solution was poured
into 10% NaHCO.sub.3 and the aqueous layer was re-extracted with
EtOAc. The combined organic phase was dried over anhydrous
Na.sub.2SO.sub.4 and concentrated in vacuo. The resulting residue
was purified by silica gel preparative TLC using 9:1:0.1
CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an eluent to afford 22 mg (15%)
of the title compound, m/z 414.6 (M+1).sup.+.
Example 17
N.sup.4-[(trans-4-{[(1,5-dimethyl-1H-pyrazol-4-yl)methyl]amino}cyclohexyl)-
methyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00114##
[0738] A mixture of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]-pyrimidine-2,4-diamine (100 mg, 0.23 mmol) and
1,5-dimethyl-1H-pyrazole-4-carbaldehyde (23 mg, 0.19 mmol) in
CH.sub.2Cl.sub.2 (15 mL) was stirred at room temperature for 1 h.
NaBH(OAc).sub.3 (200 mg, 0.95 mmol) was then added to the reaction
mixture and stirred for another 16 h. The reaction mixture was
diluted with 1M Na.sub.2CO.sub.3 to pH 9-10 and the organic phase
was separated. The aqueous phase was extracted with
CH.sub.2Cl.sub.2. The combined organic phase was dried over
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified by silica gel prep TLC using 98:2 CH.sub.2Cl.sub.2:MeOH as
an eluent to afford 54 mg (52%) of the title compound as a pale
yellow foam, m/z 549.7 [M+1].sup.+
[0739] The following compounds were prepared following similar
procedures as described above: [0740]
N.sup.4-[(trans-4-{[(3,5-dimethylisoxazol-4-yl)methyl]amino}cyclohexyl)me-
thyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine,
m/z 550.6 [M+1].sup.+ [0741]
N.sup.4-[(trans-4-{[(4-chloro-1-methyl-1H-pyrazol-3-yl)methyl]amino}cyclo-
hexyl)methyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-d-
iamine, m/z 569.6 [M+1].sup.+ [0742]
5-nitro-N.sup.4-({trans-4-[(pyrimidin-5-ylmethyl)amino]cyclohexyl}methyl)-
-N.sup.2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine, m/z
533.6 [M+1].sup.+ [0743]
N.sup.4-[(trans-4-{[(1,2-diethyl-1H-imidazol-4-yl)methyl]amino}cyclohexyl-
)methyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamin-
e, m/z 575.6 [M+1].sup.+ [0744]
N.sup.4-[(trans-4-{[(3,5-dichloropyridin-4-yl)methyl]amino}cyclohexyl)met-
hyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine,
m/z 600.6 [M+1].sup.+ [0745]
5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]-N.sup.4-{[trans-4-({[(triflu-
oromethyl)pyridin-3-yl]methyl}-amino)cyclohexyl]methyl}pyrimidine-2,4-diam-
ine, m/z 600.7 [M+1].sup.+ [0746]
5-nitro-N.sup.4-({trans-4-[(quinolin-3-ylmethyl)amino]cyclohexyl}methyl)--
N.sup.2-[2-trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine, m/z
582.5 [M+1].sup.+ [0747]
5-nitro-N.sup.4-({trans-4-[(pyridin-2-ylmethyl)amino]cyclohexyl}methyl)-N-
.sup.2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine, m/z
532.6 [M+1].sup.+ [0748]
N.sup.4-({trans-4-[(2-fluorobenzyl)amino]cyclohexylmethyl}-5-nitro-N.sup.-
2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine, m/z 549.5
[M+1].sup.+ [0749]
N.sup.4-({trans-4-[(3-fluorobenzyl)amino]cyclohexyl}methyl)-5-nitro-N.sup-
.2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine, m/z 549.6
[M+1].sup.+ [0750]
N.sup.4-({trans-4-[(4-fluorobenzyl)amino]cyclohexyl}methyl)-5-nitro-N.sup-
.2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine, m/z 549.6
[M+1].sup.+ [0751]
5-nitro-N.sup.4-({trans-4-[(quinolin-4-ylmethyl)amino]cyclohexyl}methyl)--
N.sup.2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine, m/z
582.6 [M+1].sup.+ [0752]
5-nitro-N.sup.4-({trans-4-[(2-phenylethyl)amino]cyclohexyl}methyl)-N.sup.-
2-[2-(trifluoromethoxy)-benzyl]pyrimidine-2,4-diamine, m/z 545.6
[M+1].sup.+. [0753]
2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-y-
l)amino]methyl}cyclohexyl)amino]propane-1,3-diol, m/z 515.8
[M+1].sup.+. [0754]
N.sup.4-[(trans-4-{[(1-methyl-1H-pyrazol-4-yl)methyl]amino}cyclohe-
xyl)methyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-dia-
mine, m/z 535.6 [M+1].sup.+. [0755]
N.sup.4-[(trans-4-{[(1,3-dimethyl-1H-pyrazol-4-yl)methyl]amino}cyclohexyl-
)methyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamin-
e, m/z 549.6 [M+1].sup.+. [0756]
N.sup.4-[(trans-4-{[(1,3-dimethyl-1H-pyrazol-5-yl)methyl]amino}cyclohexyl-
)methyl]-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamin-
e, m/z 549.6 [M+1].sup.+. [0757]
N.sup.4-[(trans-4-{[(1,5-dimethyl-1H-pyrazol-4-yl)methyl]amino}cyclohexyl-
)methyl]-5-nitro-{[4-(trifluoromethyl)pyridin-3-yl]methyl}pyrimidine-2,4-d-
iamine, m/z 534.5 [M+1].sup.+ [0758]
2-{[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4--
yl)amino]methyl}cyclohexyl)amino]methyl}benzoic acid, (BI0061120)
m/z 575 [M+1].sup.+
Example 18
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-5-chloro-N.sup.2-[2-(tri-
fluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00115##
[0760]
N.sup.4-[trans-4-aminocyclohexyl)methyl]-5-chloro-N.sup.2-[2-(trifl-
uoromethoxy)benzyl]pyrimidine-2,4-diamine (178 mg, 0.41 mmol) was
dissolved in DMA (5 mL). To this solution was added
1,3-dibromopropane (208 .mu.L, 2.01 mmol) and NaCO.sub.3 (217 mg,
2.01 mmol). The reaction was heated in the microwave at 100.degree.
C. for 5 min. The volatiles were removed and the reaction was
diluted with EtOAc and poured into H.sub.2O. The aqueous phase was
separated and extracted two more times with EtOAc. The organic
layers were combined, dried (Na.sub.2SO.sub.4), decanted and
concentrated. The crude product was purified on a 2000 micron
SiO.sub.2 preparative TLC plate, eluting with 10:1
CH.sub.2Cl.sub.2:MeOH (1% NH.sub.4OH) to afford 102 mg (53%) of the
title compound, m/z 470.5 [M+1].sup.+.
[0761] The following compounds were prepared following similar
procedures as described above: [0762]
5-chloro-N.sup.4-[(trans-4-pyrrolidin-1-ylcyclohexyl)methyl]-N.sup.2-[2-(-
trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 484.6
[M+1].sup.+ [0763]
1-(trans-4-{[(5-chloro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimi-
din-4-yl)amino]methyl}cyclohexyl)azetidin-3-ol, m/z 486.6
[M+1].sup.+
Example 19
5-fluoro-N.sup.4-[(trans-4-pyrrolidin-1-ylcyclohexyl)methyl]-N.sup.2-[2-(t-
rifluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00116##
[0765]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-fluoro-N.sup.2-[2-(trif-
luoromethoxy)benzyl]pyrimidine-2,4-diamine (71 mg, 0.17 mmol) was
dissolved in DMA (2 mL). To this solution was added
1,4-dibromobutane (81 .mu.L, 0.68 mmol) and NaCO.sub.3 (72 mg, 0.68
mmol). The reaction was heated in the microwave at 100.degree. C.
for 20 min. The volatiles were removed and the reaction was diluted
with EtOAc and poured into H.sub.2O. The aqueous phase was
separated and extracted two more times with EtOAc. The organic
layers were combined, dried (Na.sub.2SO.sub.4), decanted and
concentrated. The crude product was purified by twice running on a
1000 micron SiO.sub.2 Prep TLC plate, eluting with 10:1
CH.sub.2Cl.sub.2:MeOH (1% NH.sub.4OH) to afford 36 mg (45%) of the
title compound, m/z 468.6 [M+1].sup.+.
[0766] The following compounds were prepared following similar
procedures as described above: [0767]
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-5-fluoro-N.sup.2-[2-(tr-
ifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 454.5 [M+1].sup.+
[0768]
1-(trans-4-{[(5-fluoro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-y-
l)amino]methyl}-cyclohexyl)azetidin-3-ol, m/z 470.5 [M+1].sup.+
[0769]
1-[trans-4-({[5-nitro-2-({[2-(2,2,2-trifluoroethoxy)pyridin-3-yl]methyl}a-
mino)pyrimidin-4-yl]amino}methyl)cyclohexyl]azetidin-3-ol, m/z
512.5 [M+1].sup.+. [0770]
N.sup.4-[(trans-4-azetidin-1-ylcyclohexyl)methyl]-5-nitro-N.sup.2-{[2-(2,-
2,2-trifluoroethoxy)pyridin-3-yl]methyl}pyrimidine-2,4-diamine, m/z
496.5 [M+1].sup.+.
Example 20
N.sup.2-[5-chloro-2-(trifluoromethoxy)benzyl]-5-nitro-N.sup.4-[(trans-4-py-
rrolidin-1-ylcyclohexyl)methyl]pyrimidine-2,4-diamine
##STR00117##
[0772] To a solution of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[5-chloro-2-(trifluorom-
ethoxy)benzyl]-5-nitropyrimidine-2,4-diamine (51 mg, 0.1 mmol) in
DMF (0.5 mL) was added 1,4-dibromobutane (36 .mu.L, 0.3 mmol) and
DIPEA (104 .mu.L, 0.6 mmol). The reaction mixture was stirred at
50.degree. C. for 24 h and then 72 h at room temperature. The
reaction mixture was then partitioned between EtOAc (20 mL) and
saturated NaHCO.sub.3 (less than 1 mL). The organic phase was
washed with water (2.times.2 mL) and dried over Na.sub.2SO.sub.4.
The resulting residue was purified by preparative HPLC to afford 13
mg (25%) of the title compound as a yellow oily solid, m/z 529.3
[M+1].sup.+
Example 21
N.sup.2-(trans-4-{[(5-cyan-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin--
4-yl)amino]methyl}cyclohexyl)glycinamide
##STR00118##
[0774] A mixture of 2-bromoacetamide (30 mg, 0.21 mmol),
4-{[(trans-4-aminocyclohexyl)methyl]amino}-2-{[2-(trifluoromethoxy)benzyl-
]amino}pyrimidine-5-carbonitrile (117 mg, 0.28 mmol) and DIPEA (74
mL, 0.43 mmol) in DMF (1 mL) was stirred at 50.degree. C. for 4 h.
The reaction mixture was then diluted with EtOAc (30 mL) and washed
with water (3.times.3 mL) and brine. The organic phase was dried
over Na.sub.2SO.sub.4 and concentrated. Silica gel preparative TLC
of the crude product using 10:1:0.05
CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an eluent afforded 28 mg (27%)
of the title compound as a white solid, m/z 476.5 [M-1].sup.+
[0775] The following compound was prepared following similar
procedures as described above: [0776]
N.sup.2-(2-amino-2-oxoethyl)-N.sup.2-(trans-4-{[(5-cyano-2-{[2-(trifluoro-
methoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}cyclohexyl)glycinamide,
m/z 533.5 [M-1].sup.+
Example 22
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)-
amino]methyl}cyclohexyl)azetidin-3-one
##STR00119##
[0778] DMSO (170 .mu.L, 2.4 mmol) was added dropwise to a solution
of oxalyl chloride (105 .mu.L, 1.2 mmol) in dry CH.sub.2Cl.sub.2
(10 mL) cooled in a dry ice/acetone bath. The reaction mixture was
stirred with cooling for 30 min. A solution of
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)azetidin-3-ol (200 mg, 0.4 mmol) in dry
CH.sub.2Cl.sub.2 was added dropwise and the reaction was allowed to
stir for 2 h. Et.sub.3N (501 .mu.L, 3.6 mmol) was then added and
the reaction mixture was stirred for 16 h and allowed to slowly
warm to room temperature. The mixture was diluted with
CH.sub.2Cl.sub.2 and poured into saturated aqueous
Na.sub.2SO.sub.4. The aqueous phase was separated and extracted
with CH.sub.2Cl.sub.2 (.times.2). The combined organic phase was
dried over Na.sub.2SO.sub.4, decanted and concentrated. The crude
product was purified by flash chromatography on a 120 g SiO.sub.2
column, eluting with 90:9:1 CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an
eluent to afford 147 mg (74%) of the title compound, m/z 495.6
[M+1].sup.+.
Example 23
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)-
amino]methyl}cyclohexyl)azetidin-3-yl acetate
##STR00120##
[0780]
1-(trans-4-{[(5-Nitro-2-{[2-trifluoromethoxy)benzyl]amino}pyrimidin-
-4-yl)amino]methyl}cyclohexyl)azetidin-3-ol (25 mg, 0.05 mmol) was
dissolved in pyridine (12 .mu.L, 0.15 mmol) and acetic anhydride
(14 .mu.L, 0.15 mmol) was added to the solution. The reaction was
allowed to stir at 20.degree. C. for 18 h. The volatiles were
removed in vacuo and the crude product was purified by flash
chromatography on a 12 g SiO.sub.2 column, eluting with 90:9:1
CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an eluent to afford 5 mg of the
title compound, ml 539.6 [M+1].sup.+
Example 24
2-methyl-1-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrim-
idin-4-yl)amino]-methyl}cyclohexyl)amino]propan-2-ol
##STR00121##
[0782] A reaction mixture of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]pyrimidine-2,4-diamine (50 mg, 0.11 mmol) and
1,2-epoxy-2-methylpropanol (500 .mu.L) in ethanol (1 mL) was heated
to 100.degree. C. for 1 h in a microwave. The reaction mixture was
concentrated and the resulting residue was purified by silica gel
preparative TLC using 10:1 CH.sub.2Cl.sub.2:MeOH as an eluent to
afford 28 mg (47%) of the title compound as a white solid, m/z
513.5 [M+1].sup.+
Example 25
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)-
amino]methyl}cyclohexyl)azetidine-2-carboxamide
##STR00122##
[0784] To a solution of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]-pyrimidine-2,4-diamine (300 mg, 0.69 mmol) in DMF (3
mL) were added t-butyl 2,4-dibromobutyrate (322 .mu.L, 1.5 mmol)
and DIPEA (297 .mu.L), 1.7 mmol). The reaction mixture was stirred
at 60.degree. C. for 9 h. The reaction mixture was then diluted
with EtOAc and washed with 1M Na.sub.2CO.sub.3, water (.times.4)
and brine. The organic phase was then dried over Na.sub.2SO.sub.4
and concentrated. The resulting residue was purified by silica gel
preparative TLC using 98:2 CH.sub.2Cl.sub.2:MeOH as an eluent to
afford 116 mg of
1-trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)-
amino]methyl}cyclohexyl)azetidine-2-carboxylic acid tert-butyl
ester.
[0785] To a solution of
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)azetidine-2-carboxylic acid tert-butyl
ester (110 mg, 0.19 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added TFA
(3 mL). The reaction mixture was stirred at room temperature for 16
h. The mixture was concentrated and the resulting residue was
purified by silica gel preparative TLC using 4:1
CH.sub.2Cl.sub.2:MeOH as an eluent to afford the acid.
[0786] To a solution of
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)azetidine-2-carboxylic acid (170 mg, 0.27
mmol) in DMF (1.5 mL) were added ammonium chloride (170 mg, 3.2
mmol), HATU (303 mg, 0.80 mmol) and DIPEA (370 .mu.L 2.1 mmol). The
reaction mixture was stirred at room temperature for 16 h. The
reaction mixture was diluted with EtOAc and washed with saturated
NaHCO.sub.3, water (.times.3) and brine. The organic phase was
dried over Na.sub.2SO.sub.4 and concentrated. The resulting residue
was purified by silica gel preparative TLC using 95:5
CH.sub.2Cl.sub.2:MeOH as an eluent to afford 7 mg of the title
compound, m/z 522.6 [M-1].sup.+
Example 26
4-[(trans-4-aminocyclohexyl)methoxy]-5-nitro-N-[2-(trifluoromethoxy)-benzy-
l]pyrimidin-2-amine
##STR00123##
[0788] To a suspension of NaH (7 mg, 0.18 mmol, mineral oil was
removed by triturating with hexanes) in THF (2 mL) at 0.degree. C.
was added a solution of the alcohol (37 mg, 0.16 mmol) in THF (1
mL) dropwise via syringe. The cloudy reaction mixture was stirred
for 15 min, and the isocyanate (51 mg, 0.14 mmol) was added as a
THF solution dropwise via syringe. The resulting clear orange
reaction mixture was stirred at 0.degree. C. for 2 h, then warmed
to room temperature and stirred for an additional 18 h. The
reaction mixture was partitioned between EtOAc (30 mL) and water
(10 mL), and some NH.sub.4Cl was added. The organic phase was
washed with brine (10 mL), dried over Na.sub.2SO.sub.4, filtered
and concentrated. The resulting orange oil was purified by flash
chromatography (12 g of silica gel) eluting with 5-20%
EtOAc/hexanes to give 20 mg (27%) of product as a yellow oil.
[0789] The above product was dissolved in CH.sub.2Cl.sub.2 (2 mL)
and TFA (150 .mu.L) was added. The reaction mixture was stirred at
room temperature for 2 h and concentrated. The resulting residue
was purified by silica gel preparative TLC using 9:1:0.1
CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an eluent to afford 5 mg (31%)
of the title compound, m/z 442.6 [M+H].sup.+.
Example 27
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)-
amino]methyl}cyclohexyl)-2-pyridin-4-ylacetamide
##STR00124##
[0791] A mixture of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]-pyrimidine-2,4-diamine (50 mg, 0.11 mmol),
4-pyridylacetic acid hydrochloride (30 mg, 0.17 mmol), TBTU (56 mg,
0.17 mmol), and DIPEA (69 .mu.L, 0.40 mmol) in a mixture of
CH.sub.2Cl.sub.2 (1 mL) and DMF (0.1 mL) was stirred at room
temperature for 16 h. The reaction mixture was then diluted with
CH.sub.2Cl.sub.2 and washed with 1M Na.sub.2CO.sub.3 and water. The
organic phase was dried over Na.sub.2SO.sub.4 and concentrated. The
resulting residue was purified by silica gel preparative TLC using
95:5 CH.sub.2Cl.sub.2;MeOH as an eluent to afford 35 mg (55%) of
the title compound as a white solid after washing the obtained
product with diethyl ether, m/z 558.4 [M-1].sup.+
[0792] The following compounds were prepared following similar
procedures as described above: [0793]
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)-2-pyridin-3-ylacetamide, m/z 558.6
[M-1].sup.+ [0794]
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimid-
in-4-yl)amino]methyl}-cyclohexyl)-2-pyridin-2-ylacetamide, m/z
558.4 [M-1].sup.+ [0795]
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)-2-phenylacetamide, m/z 559.5[M+1].sup.+
[0796]
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
amino]methyl}-cyclohexyl)acetamide, m/z 481.3 [M-1].sup.+ [0797]
2-Hydroxy-N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyri-
midin-4-yl)amino]-methyl}cyclohexyl)acetamide, m/z 497.7
[M-1].sup.+ [0798]
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimid-
in-4-yl)amino]methyl}-cyclohexyl)benzamide, m/z 545.6 [M+1].sup.+
[0799]
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)nicotinamide, m/z 544.5 [M-1].sup.+
[0800]
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidinyl)am-
ino]methyl}-cyclohexyl)isonicotinamide, m/z 544.5 [M-1].sup.+
[0801]
N-(4-{[2-(3-Bromo-2-methyl-benzylamino)-5-nitro-pyrimidin-4-ylamino]-meth-
yl}-cyclohexyl)acetamide, m/z 492.4 [M+1].sup.+
Example 28
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)-
amino]methyl}cyclohexyl)methanesulfonamide
##STR00125##
[0803] To a solution of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]-pyrimidine-2,4-diamine (50 mg, 0.11 mmol) were added
methanesulfonyl chloride (18 .mu.L, 0.23 mmol) and DIPEA (59 .mu.L,
0.34 mmol) in CH.sub.2Cl.sub.2 (2 mL). The reaction mixture was
stirred at room temperature for 15 min and diluted with
CH.sub.2Cl.sub.2. The organic phase was then washed with 1M
Na.sub.2CO.sub.3 and water. The organic phase was dried over
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified by silica gel preparative TLC using 98:2
CH.sub.2Cl.sub.2;MeOH as an eluent to afford 28 mg (47%) of the
title compound, m/z 517.7 [M-1].sup.+
[0804] The following compounds were prepared following similar
procedures as described above: [0805]
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)-1-phenylmethanesulfonamide, m/z 596.0
[M+1].sup.+ [0806]
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)benzenesulfonamide, m/z 579.5 [M-1].sup.+
[0807]
N-[1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-
-yl)amino]methyl}cyclohexyl)azetidin-3-yl]methanesulfonamide, m/z
574.3 [M+1].sup.+ [0808]
N-{1-[trans-4-({[5-nitro-2-{[4-(trifluoromethyl)pyridin-3-yl]methyl}amino-
)pyrimidin-4-yl]amino}methyl)cyclohexyl]azetidin-3-yl}methanesulfonamide,
m/z 559.63 (M+1).sup.+. [0809]
N-[1-(Trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-
-yl)amino]methyl}cyclohexyl)azetidin-3-yl]benzenesulfonamide, m/z
636.37 (M+1).sup.+.
Example 29
N-ethyl-N'-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimi-
din-4-yl)amino]methyl}cyclohexyl)urea
##STR00126##
[0811] To a solution of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]-pyrimidine-2,4-diamine (100 mg, 0.23 mmol) in DMF (2
mL) was added ethyl isocyanate (65 mg, 0.91 mmol). The reaction
mixture was stirred at room temperature for 16 h. The resulting
precipitate was filtered and triturated in MeOH. The solid was
dried in a vacuum oven to afford 47 mg (41%) of the title compound,
m/z 512.7 [M+1].sup.+
Example 30
N-(4-{[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)a-
mino]methyl}phenyl)acetamide
##STR00127##
[0813] To a solution of
{4-[(2-(4-aminobenzyl)-5-nitro-pyrimidin-4-ylamino)-methyl]cyclohexyl}-ca-
rbamic acid tert-butyl ester (100 mg, 0.21 mmol) in
CH.sub.2Cl.sub.2 (5 mL) was cooled to 0.degree. C. To this solution
were added acetyl chloride (18 mg, 0.23 mmol) and
diisopropylethylamine (0.55 mL, 0.32 mmol). The reaction mixture
was stirred at 0.degree. C. for 10 min and an ice bath was removed.
The reaction mixture was stirred for another 30 min. The reaction
mixture was then washed with saturated NaHCO.sub.3. The organic
phase was dried over Na.sub.2SO.sub.4 and concentrated. The
resulting residue was purified by silica gel prep TLC using 95:5
CH.sub.2Cl.sub.2:MeOH as an eluent to afford
(4-{[2-(4-acetylamino-benzylamino)-5-nitro-pyrimidin-4-ylamino]-methyl}-c-
yclohexyl)-carbamic acid tert-butyl ester. This product was
dissolved in CH.sub.2Cl.sub.2 (5 mL) and TFA (1 mL) was added. The
reaction mixture was stirred at room temperature for 2 h and then
treated with 1M Na.sub.2CO.sub.3. The mixture was then extracted
with CH.sub.2Cl.sub.2. MeOH was added to the mixture during
extraction to solubilize the product into the organic phase. The
combined organic phase was dried over Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by silica gel prep
TLC using 10:1:0.2 CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH as an eluent to
afford 20 mg of the title compound as a pale yellow solid (21%),
m/z 414.5 [M+1].sup.+.
Example 31
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(2-isopropoxypyridin-3--
yl)methyl]-5-nitropyrimidine-2,4-diamine
##STR00128##
[0815] A mixture of 2-chloronicotinonitrile (1.0 g, 7.2 mmol),
2-propanol (0.87 g, 14.4 mmol), KOH (0.81 g, 14.4 mmol) and
18-crown-6 (1.8 g, 2.89 mmol) in toluene (30 mL) was stirred at
60.degree. C. for 16 h. The reaction mixture was quenched with
water and the organic phase was separated and then washed with
water (.times.5). The organic phase was dried over Na.sub.2SO.sub.4
and concentrated to yield crude 2-isopropoxy-nicotinonitrile (1.1
g, 94%) which was used without further purification.
[0816] 2-Isopropoxy-nicotinonitrile (200 mg, 1.2 mmol) obtained
above was dissolved in MeOH (5 mL) and slurry of Pd/C (wet, 50%
water content, 200 mg) in MeOH was added to the solution. The
reaction mixture was degassed using in-house vacuum and saturated
with H.sub.2 balloon. The reaction mixture was stirred at room
temperature for 6 h. The mixture was then filtered through a pad of
celite and rinsed with MeOH. The filtrate was concentrated to yield
crude (2-isopropoxy-pyridin-3-yl)methylamine which was used in the
next reaction without further purification (180 mg, 88%).
[0817] To a solution of
{4-[(2-chloro-5-nitro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}-carbamic
acid tert-butyl ester (150 mg, 0.39 mmol) in DMF (1 mL) were added
a solution of (2-isopropoxy-pyridin-3-yl)-methylamine (180 mg, 1.1
mmol) obtained above in DMF (1 mL) followed by DIPEA (135 .mu.L,
0.78 mmol). The reaction mixture was stirred at room temperature
for 30 min. The reaction mixture was diluted with ethyl acetate and
washed with saturated NaHCO.sub.3 and water (.times.4). The organic
phase was dried over Na.sub.2SO.sub.4 and concentrated. The
resulting residue was purified by silica gel prep TLC using 95:5
CH.sub.2Cl.sub.2:MeOH as an eluent to afford
[4-({2-[(2-isopropoxy-pyridin-3-ylmethyl)-amino]-5-nitro-pyrimidin-
-4-ylamino}-methyl)-cyclohexyl]-carbamic acid tert-butyl ester
which was dissolved in CH.sub.2Cl.sub.2 and TFA was added. The
reaction mixture was stirred at room temperature for 2 h. The
mixture was then treated with 1M Na.sub.2CO.sub.3 and extracted
with CH.sub.2Cl.sub.2. MeOH was added to the mixture during the
extraction to solubilize the product into the organic phase. The
combined organic phase was dried over Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by silica gel prep
TLC using 10:1:0.2 CH.sub.2Cl.sub.2;MeOH:NH.sub.4OH as an eluent to
afford 75 mg of the title compound as a pale yellow solid (46%);
m/z 416.4 [M+1].sup.+.
[0818] The following compounds were prepared using similar
procedures as described above: [0819]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(2-ethoxypyridin-3-yl)-
methyl]-5-nitropyrimidine-2,4-diamine, m/z 400.2 [M-1].sup.+.
[0820]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-{[2-(2,2,2-trif-
luoroethoxy)pyridin-3-yl]methyl}pyrimidine-2,4-diamine, m/z 456.4
[M+1].sup.+.
Example 32
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(5-fluoro-2-methoxypyri-
din-3-yl)methyl]-5-nitropyrimidine-2,4-diamine
##STR00129##
[0822] 1,8-Diazabicyclo[5.4.0]undec-7-ene (630 mg, 4.1 mmol) was
added to a solution of 5-fluoro-3-hydroxymethyl-2-methoxypyridine
(500 mg, 3.2 mmol) and diphenylphosphoryl azide (1050 mg, 3.8 mmol)
in toluene (10 mL) at room temperature. The reaction mixture was
stirred at room temperature for 3 h. The reaction mixture was then
diluted with 1M HCl and ether. The layers were separated and the
aqueous layer was extracted with ether (.times.2). The combined
organic phase was washed with water, brine and dried over
Na.sub.2SO.sub.4. The resulting residue was purified by silica gel
prep TLC using CH.sub.2Cl.sub.2 as an eluent to afford 320 mg of
3-azidomethyl-5-fluoro-2-methoxy-pyridine as colorless oil
(55%).
[0823] To a solution of 3-azidomethyl-5-fluoro-2-methoxy-pyridine
(320 mg, 1.8 mmol) in MeOH (8 mL) was added slurry of Pd/C (wet,
50% water content, 100 mg) in MeOH (2 mL). The reaction mixture was
degassed using in-house vacuum for 5 min and then H.sub.2 balloon
was attached to the reaction flask. The reaction mixture was
stirred at room temperature for 2 h. The mixture was then filtered
through a pad of celite and the filtrated was concentrated to
afford 280 mg of (5-fluoro-2-methoxy-pyridin-3-yl)-methylamine
(100%). This product was used for the next reaction without further
purification.
[0824] To a solution of
{4-[(2-chloro-5-nitro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}-carbamic
acid tert-butyl ester (200 mg, 0.52 mmol) in DMF (2 mL) were added
a solution of (5-fluoro-2-methoxy-pyridin-3-yl)-methylamine (120
mg, 0.78 mmol) in DMF (1 mL) followed by diisopropylethylamine (181
.mu.L, 1.04 mmol). The reaction mixture was stirred at room
temperature for 30 min. The reaction mixture was diluted with ethyl
acetate and washed with saturated NaHCO.sub.3 and water (.times.4).
The organic phase was dried over Na.sub.2SO.sub.4, filtered and
concentrated. The resulting residue was purified by silica gel prep
TLC using 95:5 CH.sub.2Cl.sub.2:MeOH as an eluent to afford
[4-({2-[(5-fluoro-2-methoxy-pyridin-3-ylmethyl)-amino]-5-nitro-pyrimidin--
4-ylamino}-methyl)-cyclohexyl]-carbamic acid tert-butyl ester which
was then dissolved in CH.sub.2Cl.sub.2 (5 mL) and TFA (1 mL) was
added. The reaction mixture was stirred at room temperature for 2
h. The mixture was then treated with 1M Na.sub.2CO.sub.3 and
extracted with CH.sub.2Cl.sub.2. MeOH was added to the mixture
during the extraction to solubilize the product into the organic
phase. The combined organic phase was dried over Na.sub.2SO.sub.4
and concentrated. The resulting residue was purified by silica gel
prep TLC using 10:1:0.2 CH.sub.2Cl.sub.2;MeOH:NH.sub.4OH as an
eluent to afford 156 mg of the title compound as a pale yellow
solid (74%), m/z 406.4 [M+1].sup.+.
Example 33
N.sup.4-{[trans-4-(3-fluoroazetidin-1-yl)cyclohexyl]methyl}-5-nitro-N.sup.-
2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00130##
[0826] To a solution of
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)azetidin-3-ol (100 mg, 0.20 mmol) in
CH.sub.2Cl.sub.2 (25 mL) was added dropwise a solution of
diethylaminosulfur trifluoride (32 .mu.L, 0.24 mmol) in
CH.sub.2Cl.sub.2 (0.5 mL) at room temperature over a period of 15
min. When the reaction was complete, the mixture was treated with
saturated NaHCO.sub.3 and stirred for 5 min. The organic phase was
separated and washed with water. The organic phase was dried over
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified by silica gel prep TLC using 95:5 CH.sub.2Cl.sub.2:MeOH as
eluent to afford 26 mg of the title compound as a pale yellow solid
(26%), m/z 499.1 [M+1].sup.+.
Example 34
5-nitro-N.sup.4-({trans-4-[(2,2,2-trifluoroethyl)amino]cyclohexyl}methyl)--
N.sup.2-[2-trifluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00131##
[0828]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifl-
uoromethoxy)benzyl]pyrimidine-2,4-diamine (200 mg, 0.45 mmol) was
suspended in CH.sub.3CN (4 mL) and cooled to 0.degree. C.
2,2,2-Trifluoroethyl trichloromethanesulphonate (66 mg, 0.23 mmol)
in CH.sub.3CN (1 mL) was added dropwise to the reaction mixture and
the reaction mixture was heated to 90.degree. C. for 5 h. The
reaction mixture was concentrated and the resulting residue was
diluted with ethyl acetate. The solution was treated with 1M
Na.sub.2CO.sub.3 and the organic phase was separated. The aqueous
phase was re-extracted with CH.sub.2Cl.sub.2. The combined organic
phase was dried over Na.sub.2SO.sub.4 and concentrated. Silica gel
prep TLC using 98:2 CH.sub.2Cl.sub.2:MeOH, then 7:3 Hexanes:ethyl
acetate afforded 74 mg of the title compound, m/z 523.0
[M+1].sup.+.
Example 35
(1S)-2-[(trans-4-{[5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin--
4-yl)amino]methyl}cyclohexyl)amino]-1-pyridin-3-ylethanol
##STR00132##
[0830] (S)-2-Methyl-CBS-oxazaborolidine (1M in toluene, 189 .mu.L,
0.18 mmol) and borane-methyldisulfide complex (2M in toluene, 100
.mu.L) in THF (11 mL) was allowed to stir at room temperature for
10 min. Then, the rest of BH.sub.3--S(CH.sub.3).sub.2 (2M in
toluene, 1.33 mL, 2.85 mmol) was added to the reaction solution
followed by a suspension of 3-bromoacetyl)pyridine hydrochloride
(0.5 g, 1.78 mmol) in THF (11 mL). The heterogeneous reaction
mixture was stirred at room temperature for 16 h. The reaction
mixture was then cooled to 0.degree. C. and quenched with MeOH (1.5
mL). After 5 min of stirring, the solvent was removed and the
resulting residue was diluted with CH.sub.2Cl.sub.2. The solution
was treated with saturated NaHCO.sub.3 and the organic phase was
separated. The aqueous phase was re-extracted with
CH.sub.2Cl.sub.2. The combined organic phase was dried over
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified by silica gel prep TLC using 95:5 CH.sub.2Cl.sub.2:MeOH as
an eluent to afford 0.24 g of (S)-2-bromo-1-pyridin-3-yl-ethanol as
a colorless oil (66%).
[0831] To a solution of (S)-2-Bromo-1-pyridin-3-yl-ethanol (134 mg,
0.66 mmol) in THF (10 mL) was added 4N NaOH solution (8.3 mL). The
reaction mixture was stirred at room temperature for 1 h. The
organic phase was separated, dried over Na.sub.2SO.sub.4 and
concentrated. Silica gel prep TLC of the resulting residue using
99:1 CH.sub.2Cl.sub.2;MeOH as an eluant afforded 34 mg of
(S)-3-Oxiranyl-pyridine as a colorless oil (42%).
[0832] A mixture of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]pyrimidine-2,4-diamine (124 mg, 0.28 mmol),
(S)-3-oxiranyl-pyridine (34 mg, 0.28 mmol) and lithium perchlorate
(60 mg, 0.56 mmol) in CH.sub.3CN was heated to 100.degree. C. in a
sealed tube and stirred for 1 h. The reaction mixture was
concentrated and diluted with ethyl acetate. The solution was then
treated with 1M Na.sub.2CO.sub.3 and the organic phase was
separated. The aqueous phase was re-extracted with ethyl acetate
(.times.2). The combined organic phase was dried over
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified by silica gel prep TLC using 10:1:0.05
CH.sub.2Cl.sub.2;MeOH:NH.sub.4OH as an eluent to afford 15 mg of
the title compound as an off-white solid (10%), m/z 562.7
[M+1].sup.+.
Example 36
5-nitro-N.sup.4-{[trans-4-(pyridin-2-ylamino)cyclohexyl]methyl}-N.sup.2-[2-
-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00133##
[0834] A mixture of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[2-(trifluorome-
thoxy)benzyl]pyrimidine-2,4-diamine (100 mg, 0.23 mmol),
2-fluoropyridine (0.5 mL) and diisopropylethyl (0.2 mL) was placed
in a microwave tube and heated to 150.degree. C. in the Personal
Chemistry Microwave for 1 h, 165.degree. C. for 1 h, then
185.degree. C. for another 2 h. The reaction mixture was
concentrated and the resulting residue was diluted with ethyl
acetate. The organic phase was washed with 10% citric acid and
brine. The organic phase was dried over Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by silica gel prep
TLC using 98:2 CH.sub.2Cl.sub.2:MeOH as an eluent to afford 29 mg
of the title compound, m/z 518.9 [M+1].sup.+.
Example 37
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(isoquinolin-1-ylmethyl)-
-5-nitropyrimidine-2,4-diamine
##STR00134##
[0836] 1-Isoquinolinecarbonitrile (2.0 g, 12.8 mmol) was dissolved
in MeOH (160 mL). To this solution, was added 6 pasteur pipet full
of Raney Nickel suspension in water. The reaction flask was
degassed and backflushed with N.sub.2 twice. The N.sub.2 balloon
was replaced with H.sub.2 balloon and the flask was filled with
H.sub.2 after evacuation of N.sub.2. The reaction mixture was
stirred for 5 h and then filtered through a pad of celite under
stream of N.sub.2. The celite pad was flushed with MeOH. The
combined organic solution was concentrated to afford 1.32 g of
isoquinolin-1-yl-methylamine as brown oil (65%).
[0837] To a solution of isoquinolin-1-yl-methylamine (1.32 g, 8.35
mmol) and diisopropylethylamine (2.7 mL, 15.5 mmol) in DMF (20 mL)
was added
{4-[(2-chloro-5-nitro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}-carbamic
acid tert-butyl ester (1.5 g, 3.89 mmol). The reaction mixture was
stirred at room temperature for 10 min. The solvent was removed and
the resulting residue was diluted with a mixture of ethyl acetate
(350 mL) and CH.sub.2Cl.sub.2 (100 mL). The solution was then
washed with 1M Na.sub.2CO.sub.3 and water. The organic phase was
dried over Na.sub.2SO.sub.4 and concentrated. The resulting solid
was treated with ethyl acetate (poor solubility of the product in
ethyl acetate) and the un-dissolved solid was filtered through a
Buchner funnel. The white solid was dried in vacuo to yield 1.64 g
of
[4-({2-[(isoquinolin-1-ylmethyl)-amino]-5-nitro-pyrimidin-4-ylamino}-meth-
ylcyclohexyl]-carbamic acid tert-butyl ester.
[0838]
[4-({2-[(Isoquinolin-1-ylmethyl)-amino]-5-nitro-pyrimidin-4-ylamino-
}-methyl)-cyclohexyl]-carbamic acid tert-butyl ester (1.64 g, 3.24
mmol) was dissolved in CH.sub.2Cl.sub.2 (50 mL) and TFA (15 mL) was
added. The reaction mixture was stirred at room temperature for 30
min. The reaction mixture was treated with 1M Na.sub.2CO.sub.3 and
the organic phase was separated. The organic phase was dried over
Na.sub.2SO.sub.4 and concentrated. Silica gel purification using a
mixture of CH.sub.2Cl.sub.2:MeOH:NH.sub.4OH yielded 1.1 g of the
title compound (84%), m/z 406.2 (M-1).sup.+.
[0839] The following compound was prepared using similar procedures
as described above: [0840]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-{[4-(trifluorom-
ethyl)pyridin-3-yl]methyl}pyrimidine-2,4-diamine, m/z 426.4
(M+1).sup.+
Example 38
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(quinolin-5-ylme-
thyl)pyrimidine-2,4-diamine
##STR00135##
[0842] To a solution of quinoline-5-carboxylic acid (500 mg, 2.83
mmol) in THF (10 mL) was added slowly LiAlH.sub.4 (1.0 M, THF)
solution at 0.degree. C. The resulting residue was heated to
70.degree. C. for 3 h and stirred at room temperature for 16 h. The
reaction mixture was cooled to 0.degree. C. and quenched with water
(1 mL) and 10% NaOH (1.5 mL) solution. The reaction mixture was
stirred for 1 h. The solution was filtered through a pad of celite
and rinsed with THF. The filtrate was concentrated and the
resulting residue was purified by silica gel prep TLC using 95:5
CH.sub.2Cl.sub.2:MeOH as an eluent to afford 218 mg of
quinolin-5-yl-methanol (48%).
[0843] To a solution of quinolin-5-yl-methanol (96 mg, 0.60 mmol)
and DPPA (202 .mu.L, 0.91 mmol) in toluene (2 mL) was added DBU
(166 .mu.L, 1.09 mmol) at room temperature. The reaction mixture
was stirred at room temperature for 16 h. The reaction mixture was
diluted with 3M HCl (1.7 mL) and ethyl acetate (5 mL). The organic
phase was separated and the aqueous phase was re-extracted with
ethyl acetate (.times.2). The combined organic phase was dried over
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified by silica gel prep TLC using 98:2 CH.sub.2Cl.sub.2:MeOH as
an eluent to afford 49 mg of 5-azidomethyl-quinoline as light brown
oil (44%).
[0844] Triphenylphosphine (172 mg, 0.65 mmol) was added to the
solution of 5-azidomethyl-quinoline (109 mg, 0.59 mmol) in THF (3
mL) at 0.degree. C. After 5 min of stirring, the reaction mixture
was warmed to room temperature and stirred at that temperature 16
h. The reaction mixture was diluted with ammonium hydroxide (0.55
mL) and stirred for 3 h. The mixture was then treated with 2M NaOH
(0.8 mL) and stirred for another 1 h. The solution was diluted with
ethyl acetate and the organic phase was separated, dried over
Na.sub.2SO.sub.4, concentrated. The resulting residue was purified
by silica gel prep TLC using 10:1:0.3
CH.sub.2Cl.sub.2:MeOH:Et.sub.3N as an eluant to afford 105 mg of
quinolin-5-yl-methylamine as a colorless solid.
[0845] To a solution of quinolin-5-yl-methylamine 100 mg, 0.39
mmol) and diisopropylethylamine (140 .mu.L, 0.78 mmol) in a mixture
of DMF (2 mL) and acetonitrile (2 mL) was added
{4-[(2-chloro-5-nitro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}-carbamic
acid tert-butyl ester (150 mg, 0.39 mmol). The reaction mixture was
stirred at room temperature for 16 h. The reaction mixture was
diluted with ethyl acetate and washed with water (.times.2). The
combined organic phase was dried over Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by silica gel prep
TLC using 95:5 CH.sub.2Cl.sub.2:MeOH as an eluent to afford 197 mg
of
[4-({5-nitro-2-[(quinolin-5-ylmethyl)-amino]-pyrimidin-4-ylamino}-methyl)-
-cyclohexyl]-carbamic acid tert-butyl ester as a light yellow solid
(100%).
[0846]
[4-({5-Nitro-2-[(quinolin-5-ylmethyl)-amino]-pyrimidin-4-ylamino}-m-
ethyl)cyclohexyl]-carbamic acid tert-butyl ester (197 mg, 0.39
mmol) was dissolved in CH.sub.2Cl.sub.2 (10 mL) and TFA (2 mL) was
added. The reaction mixture was stirred at room temperature for 1
h. The reaction mixture was treated with 3M Na.sub.2CO.sub.3 and
the organic phase was separated. The organic phase was dried over
Na.sub.2SO.sub.4 and concentrated. Silica gel purification using a
mixture of 10:1:0.3 CH.sub.2Cl.sub.2:MeOH:Et.sub.3N yielded 120 mg
of the title compound as a light yellow solid (79%), m/z 406.4
[M-1].sup.+.
[0847] The following compounds were prepared following similar
procedures as described above: [0848]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(3-chloropyridin-4-yl)-
methyl]-5-nitropyrimidine-2,4-diamine m/z 392.5 [M+1].sup.+ [0849]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[(2-methylpyridin-3-yl)-
methyl]-5-nitropyrimidine-2,4-diamine m/z 372.6 [M+1].sup.+
Example 39
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(2-oxo-2-piperid-
in-1-ylethyl)pyrimidine-2,4-diamine
##STR00136##
[0851] To a solution of N-(tert-butoxycarbonyl)glycine (500 mg, 2.8
mmol), piperidine (0.6 mL, 5.6 mmol) in a mixture of
CH.sub.2Cl.sub.2 (10 mL) and DMF (1 mL) were added TBTU (1.4 g, 4.2
mmol) and diisopropylethylamine (970 .mu.L). The reaction mixture
was stirred at room temperature for 16 h. The reaction mixture was
concentrated and the resulting residue was diluted with ethyl
acetate.
[0852] The organic phase was washed with water (.times.3) and
brine. The combined organic phase was dried over Na.sub.2SO.sub.4
and concentrated. Silica gel column chromatography using 98:2
CH.sub.2Cl.sub.2:MeOH as an eluent afforded 590 mg of
(2-oxo-2-piperidin-1-yl-ethyl)-carbamic acid tert-butyl as a
colorless solid (87%).
[0853] (2-Oxo-2-piperidin-1-yl-ethyl)-carbamic acid tert-butyl was
dissolved in CH.sub.2Cl.sub.2 (10 mL) and TFA (3 mL) was added to
the solution. The reaction mixture was stirred at room temperature
for 16 h. The mixture was concentrated and the resulting residue of
2-amino-1-piperidin-1-yl-ethanone was used for the next reaction
without further purification as a trifluoroacetic acid salt.
[0854] To a solution of
{4-[(2-chloro-5-nitro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}-carbamic
acid tert-butyl ester (100 mg, 0.26 mmol) in DMF (3 mL) was added
2-amino-1-piperidin-1-yl-ethanone trifluoroacetic salt (200 mg,
0.78 mmol) and diisopropylethylamine (272 .mu.L, 1.6 mmol). The
reaction mixture was stirred at room temperature for 48 h. The
mixture was diluted with ethyl acetate and washed with water
(.times.3). The organic phase was dried over Na.sub.2SO.sub.4 and
concentrated. The resulting residue was purified by silica gel prep
TLC using 95:5 CH.sub.2Cl.sub.2:MeOH as an fluent to afford 36 mg
of
(4-{[5-nitro-2-(2-oxo-2-piperidin-1-yl-ethylamino)-pyrimidin-4-ylamino]-m-
ethyl}-cyclohexyl)-carbamic acid tert-butyl ester as an off-white
solid (28%).
[0855]
(4-{[5-Nitro-2-(2-oxo-2-piperidin-1-yl-ethylamino)-pyrimidin-4-ylam-
ino]-methyl}-cyclohexyl)-carbamic acid tert-butyl ester (36 mg,
0.07 mmol) was dissolved in CH.sub.2Cl.sub.2 (10 mL) and TFA (1 mL)
was added to the solution. The reaction mixture was stirred at room
temperature for 16 h. The solution was treated with 3M
Na.sub.2CO.sub.3 to pH 9 and the organic phase was separated. The
aqueous phase was extracted with CH.sub.2Cl.sub.2 and the combined
organic phase was dried over Na.sub.2SO.sub.4 and then
concentrated. The resulting residue was purified by silica gel prep
TLC using 10:1:0.3 CH.sub.2Cl.sub.2:MeOH:Et.sub.3N as an eluent to
afford 27 mg of the title compound as a yellow solid (93%), m/z
392.6 [M+1].sup.+.
[0856] The following compounds were prepared using similar
procedures as described above: [0857]
(3R)-1-[N.sup.4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-
-2-yl)glycyl]piperidin-3-ol, m/z 406.4 [M-1].sup.+. [0858]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-morpholin-4-yl-2-oxo-
ethyl)-5-nitropyrimidine-2,4-diamine, m/z 394.5 [M+1].sup.+.
Example 40
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(2-oxo-2-pyrroli-
din-1-ylethyl)pyrimidine-2,4-diamine
##STR00137##
[0860] To a suspension of
{4-[(4-tert-butoxycarbonylamino-cyclohexylmethyl)-amino]-5-nitro-pyrimidi-
n-2-ylamino}-acetic acid methyl ester (845 mg, 1.9 mmol) in a
mixture of MeOH (90 mL) and water (30 mL) was added lithium
hydroxide monohydrate (404 mg, 9.6 mmol). The heterogeneous mixture
was stirred at room temperature for 1 h. The mixture was
concentrated and the resulting residue was acidified to pH 5 using
1M HCl. The product was extracted using ethyl acetate (.times.3).
The combined organic phase was dried over Na.sub.2SO.sub.4 and
concentrated to yield 1.0 g of the crude
{4-[(4-tert-butoxycarbonylamino-cyclohexylmethyl)-amino]-5-nitro-pyrimidi-
n-2-ylamino}-acetic acid as a white solid. This product was used
for the next reaction without further purification.
[0861] To
{4-[(4-tert-butoxycarbonylamino-cyclohexylmethyl)-amino]-5-nitro-
-pyrimidin-2-ylamino}-acetic acid (120 mg, 0.28 mmol) in DMF (1.5
mL) were added pyrrolidine (48 .mu.L, 0.57 mmol), TBTU (141 mg,
0.43 mmol), and diisopropylethylamine (99 .mu.L, 0.57 mmol). The
reaction mixture was stirred at room temperature for 16 h. The
mixture was diluted with ethyl acetate and washed with water
(.times.2) and brine. The organic phase was dried over
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified by silica gel prep TLC using 95:5 CH.sub.2Cl.sub.2:MeOH as
an eluant to afford 19 mg of
(4-{[5-nitro-2-(2-oxo-2-pyrrolidin-1-yl-ethylamino)-pyrimidin-4-ylamino]--
methyl}-cyclohexyl)-carbamic acid tert-butyl ester as a light brown
solid.
[0862]
(4-{[5-Nitro-2-(2-oxo-2-pyrrolidin-1-yl-ethylamino)-pyrimidin-4-yla-
mino]-methyl}-cyclohexyl carbamic acid tert-butyl ester (19 mg,
0.04 mmol) was dissolved in CH.sub.2Cl.sub.2 (5 mL) and TFA (1 mL)
was added. The reaction mixture was stirred at room temperature for
1 h. The solution was treated with saturated NaHCO.sub.3 and the
organic phase was separated. The aqueous phase was extracted with
CH.sub.2Cl.sub.2 and the combined organic phase was dried over
Na.sub.2SO.sub.4 and concentrated. The resulting residue was
purified by silica gel prep TLC using 10:1:0.3
CH.sub.2Cl.sub.2:MeOH:Et.sub.3N as an eluant to afford 13 mg of the
title compound as a light yellow solid (83%), m/z 376.3
[M-1].sup.+.
[0863] The following compounds were prepared using similar
procedures as described above: [0864]
1-[N-(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)gl-
ycyl]piperidin-4-ol, m/z 408.6 [M+1].sup.+. [0865]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(2-oxo-2-pipera-
zin-1-ylethyl)pyrimidine-2,4-diamine, m/z 393.6 [M+1].sup.+. [0866]
(3R)-1-[N-(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2--
yl)glycyl]pyrrolidin-3-ol, m/z 394.6 [M+1].sup.+. [0867]
1-[N-(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)gl-
ycyl]piperidine-4-carboxamide, m/z 433.3 [M-1].sup.+. [0868]
(3S)-1-[N-(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2--
yl)glycyl]pyrrolidin-3-ol, m/z 392.3 [M-1].sup.+. [0869]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(2-azetidin-1-yl-2-oxoe-
thyl)-5-nitropyrimidine-2,4-diamine, m/z 364.6 [M+1].sup.+. [0870]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[(1S)-2-oxo-1-p-
henyl-2-piperidin-1-ylethyl]pyrimidine-2,4-diamine, m/z 468.5
[M+1].sup.+. [0871]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[(1R)-2--
oxo-1-phenyl-2-piperidin-1-ylethyl]pyrimidine-2,4-diamine, m/z
468.5 [M+1].sup.+.
Example 41
N.sup.4-{[trans-4-(3-aminoazetidin-1-yl)cyclohexyl]methyl}-5-nitro-N.sup.2-
-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00138##
[0873]
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}-cyclohexyl)azetidin-3-ol (111 mg, 0.22 mmol,
see Example 7), phthalimide (50 mg, 0.33 mmol) and PPh.sub.3 (127
mg, 0.48 mmol) were combined in THF (5 mL). To this mixture was
added DEAD (61 mg, 0.35 mmol) and the reaction was allowed to stir
at 25.degree. C. for 18 h. The reaction was diluted with EtOAc and
poured into 10% aqueous NaHCO.sub.3. The aqueous phase was
separated and extracted two more times with EtOAc. The organic
layers were combined, dried (Na.sub.2SO.sub.4), decanted and
concentrated. The crude was purified via flash chromatography
(SiO.sub.2, 1:9:90-NH.sub.4OH:CH.sub.3OH:CH.sub.2Cl.sub.2) to
afford 92 mg (67%) of
2-[1-(4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylamino]-
-methyl}-cyclohexyl)-azetidin-3-yl]-isoindole-1,3-dione.
[0874]
2-[1-(4-{[5-Nitro-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-yl-
amino]-methyl}-cyclohexyl)-azetidin-3-yl]-isoindole-1,3-dione (92
mg, 0.15 mmol) was dissolved in EtOH-CH.sub.2Cl.sub.2 (3 mL, 2:1).
With stirring, hydrazine monohydrate (11.3 mg, 0.23 mmol) was added
and the reaction was stirred at 25.degree. C. for 3 h. The
volatiles were removed in vacuo and the crude residue was purified
via flash chromatography (SiO.sub.2,
1:9:90-NH.sub.4OH:CH.sub.3OH:CH.sub.2Cl.sub.2) to afford 46 mg
(62%) of the target compound, m/z 496.35 [M+1].sup.+.
Example 42
N-[1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4--
yl)amino]methyl}cyclohexyl)azetidin-3-yl]acetamide
##STR00139##
[0876]
N.sup.4-{[Trans-4-(3-aminoazetidin-1-yl)cyclohexyl]methyl}-5-nitro--
N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine (20 mg,
0.04 mmol) was dissolved in CH.sub.2Cl.sub.2 (1.0 mL). To this
solution was added acetic anhydride (0.057 mL, 0.06 mmol) followed
by triethylamine (0.084 mL, 0.06 mmol). The reaction was allowed to
stir at 25.degree. C. for 18 h. The volatiles were removed and the
crude was redissolved in CH.sub.2Cl.sub.2 and extracted with 10%
aqueous NaHCO.sub.3. The aqueous phase was separated and extracted
two more times with CH.sub.2Cl.sub.2. The organic layers were
combined, dried (Na.sub.2SO.sub.4), decanted and concentrated. The
resultant residue was purified via flash chromatography (SiO.sub.2,
10-30% (2:18:80,
NH.sub.4OH:CH.sub.3OH:CH.sub.2Cl.sub.2)--CH.sub.2Cl.sub.2) to
afford 16 mg (74%) of the target compound, m/z 538.46
(M+1).sup.+.
Example 43
N-[1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4--
yl)amino]methyl}cyclohexyl)azetidin-3-yl]benzamide
##STR00140##
[0878]
N.sup.4-{[Trans-4-(3-aminoazetidin-1-yl)cyclohexyl]methyl}-5-nitro--
N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine (20 mg,
0.04 mmol) was dissolved in THF (1.0 mL). To this solution was
added benzoic acid (7.3 mg, 0.06 mmol), Polystyrene-linked
Carbodiimide (PS-CDI) (47 mg, 0.06 mmol) followed by triethylamine
(0.084 mL, 0.06 mmol). The reaction was allowed to stir at
25.degree. C. for 18 h. The reaction was filtered to remove the
PS-CDI and the volatiles were removed in vacuo. The crude was
redissolved in CH.sub.2Cl.sub.2 and extracted with 10% aqueous
NaHCO.sub.3. The aqueous phase was separated and extracted two more
times with CH.sub.2Cl.sub.2. The organic layers were combined,
dried (Na.sub.2SO.sub.4), decanted and concentrated. The resultant
residue was purified via flash chromatography (SiO.sub.2, 10-30%
(2:18:80,
NH.sub.4OH:CH.sub.3OH:CH.sub.2Cl.sub.2)--CH.sub.2Cl.sub.2) to
afford 18 mg (74%) of the target compound, m/z 600.34
(M+1).sup.+.
Example 44
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[(1-oxidopyridin-
-2-yl)methyl]pyrimidine-2,4-diamine
##STR00141##
[0880] 3-Aminomethylpyridine (2.5 g, 23 mmol), di-tert-butyl
dicarbonate (7.72 g, 35.4 mmol) and diisopropylethyl amine (6.2 mL,
35.4 mmole) were combined in CH.sub.2Cl.sub.2 (50 mL). The reaction
was allowed to stir for 18 h. The reaction was worked up by removal
of the solvent in vacuo. The resultant residue was dissolved in
EtOAc and poured into 10% aqueous NaHCO.sub.3. The aqueous phase
was separated and extracted two more times with EtOAc. The organic
layers were combined, dried (Na.sub.2SO.sub.4), decanted and the
concentrated. The crude was purified via flash chromatography
(SiO.sub.2, 2-5% MeOH--CH.sub.2Cl.sub.2) to afford 3.9 g (81%) of
pyridin-3-ylmethyl-carbamic acid tert-butyl ester.
[0881] To pyridin-3-ylmethyl-carbamic acid tert-butyl ester (3.89
g, 18.7 mmol) dissolved in EtOH (50 mL), was added
3-chloroperoxybenzoic acid (4.7 g, 27.4 mmol). After 4 hours the
volatiles were removed in vacuo and the crude was redissolved in
CH.sub.2Cl.sub.2 (25 mL). To this solution was added TFA (25 mL)
and the reaction was stirred at 25.degree. C. for 2 h then the
volatiles were removed in vacuo. The crude was redissolved in
CH.sub.2Cl.sub.2 and evaporated. This procedure was done twice
more. (1-Oxy-pyridin-3-yl)-methylamine was obtained with
contamination of 3-chlorobenzoic acid, but carried further without
any additional purification (2.74 g).
[0882]
{trans-4-[(2-Chloro-5-nitro-pyrimidin-4-ylamino)-methyl]-cyclohexyl-
}-carbamic acid tert-butyl ester (105 mg, 0.27 mmol) was dissolved
in DMA (5 mL). To this solution was added the
(1-oxy-pyridin-3-yl)-methylamine (98 mg, 0.79 mmol), followed by
DIEA (0.14 mL, 0.79 mmol). The reaction was allowed to stir at
25.degree. C. for 18 h. The volatiles were removed in vacuo and the
crude was purified via flash chromatography (SiO.sub.2, 2-8%
MeOH--CH.sub.2Cl.sub.2).
[4-({5-Nitro-2-[(1-oxy-pyridin-3-ylmethyl)-amino]-pyrimidin-4-ylamino}-me-
thyl)-cyclohexyl]-carbamic acid tert-butyl ester was obtained pure
and was resuspended in CH.sub.2Cl.sub.2 (2.5 mL) To this suspension
was added TFA (2.5 mL) and the reaction was stirred for 2 h. The
volatiles were removed in vacuo and the crude was purified via
flash chromatography (SiO.sub.2, 25-100%
(2:18:80-NH.sub.4OH:MeOH:CH.sub.2Cl.sub.2)--CH.sub.2Cl.sub.2) to
afford 107 mg (36%) of the target compound, m/z 374.41
[M+1].sup.+.
[0883] The following compound was prepared following a procedure
similar to that described above: [0884]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-[(1-oxidopyridi-
n-2-yl)methyl]pyrimidine-2,4-diamine, m/z 374.59 [M+1].sup.+.
Example 45
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-bromo-N.sup.2-[2-(trifluoromet-
hoxy)benzyl]pyrimidine-2,4-diamine
##STR00142##
[0886] 5-Bromo-2,4-dichloropyrimidine (1.1 g, 4.83 mmol) was
dissolved in DMA (25 mL). To this solution was added
tert-butyl-trans-4-aminomethylcyclohexylcarbamate hydrochloride
(1.54 g, 5.80 mmol), followed by DIEA (1.98 mL, 11.4 mmol). The
reaction was stirred at 25.degree. C. for 18 h. The reaction was
worked up by removal of the volatiles in vacuo. The crude was
purified via flash chromatography (SiO.sub.2, 10-25% EtOAc-Hexanes)
to afford 1.32 g (65%) of
{4-[(5-bromo-2-chloro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}-carbami-
c acid tert-butyl ester.
[0887]
{4-[(2-chloro-5-nitro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}-carb-
amic acid tert-butyl ester (1.32 g, 3.14 mmol) was dissolved in
2-trifluoromethoxybenzyl amine (4.0 g, 20.9 mmol). The reaction was
heated, in two separate iterations, to 140.degree. C. for 10 min in
a Smith Synthesizer Microwave Reactor. The reaction was diluted
with CH.sub.2Cl.sub.2 and poured into 1 M aqueous HCl. The aqueous
phase was separated and extracted two more times with
CH.sub.2Cl.sub.2. The organic layers were combined, dried
(Na.sub.2SO.sub.4), decanted and concentrated. The crude was
diluted with minimal THF:MeOH:Hexane:EtOAc (25:1:49:25) and
purified via flash chromatography (SiO.sub.2, 15-25% EtOAc-Hexanes)
to afford 103 g (57%) of
(4-{[5-bromo-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylamino]-meth-
yl}-cyclohexyl)-carbamic acid tert-butyl ester.
[0888]
(4-{[5-Bromo-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylamino-
]-methyl}-cyclohexyl)-carbamic acid tert-butyl ester (75 mg, 0.13
mmol) was dissolved in CH.sub.2Cl.sub.2 (2 mL). To this solution
was added TFA (2 mL) and the reaction was stirred at 25.degree. C.
for 1 h. The volatiles were removed in vacuo. The crude was
redissolved in CH.sub.2Cl.sub.2 and poured into 10% aqueous
NaHCO.sub.3. The aqueous phase was separated and extracted two more
times with CH.sub.2Cl.sub.2. The organic layers were combined,
dried (Na.sub.2SO.sub.4), decanted and concentrated. The crude was
purified via flash chromatography (SiO.sub.2, 2-10%
(NH.sub.4OH:CH.sub.3OH:CH.sub.2Cl.sub.2-CH.sub.2Cl.sub.2)) to
afford 45 mg (73%) of the target compound, m/z 474.38
[M+1].sup.+.
Example 46
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-(phenylethynyl)-N.sup.2-[2-(tr-
ifluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00143##
[0890]
(trans-4-{[5-Bromo-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-y-
lamino]-methyl}-cyclohexyl)-carbamic acid tert-butyl ester (100 mg,
0.174 mmol), CuI (0.97 mg, 0.005 mmol), and Pd(PhCN).sub.2Cl.sub.2
(1.96 mg, 0.005 mmol) were dissolved in degassed dioxane (0.5
mmol). To this solution was added tri-(tert)butylphosphine
tetrafluoroborate (2.4 mg, 0.010 mmol) followed by diisopropylamine
(0.029 mL, 0.2 mmol) and phenylacetylene (0.022 mL, 0.2 mmol). The
reaction was stirred at 25.degree. C. for 18 h. The reaction was
worked up by the addition of EtOAc and filtered through a plug of
SiO.sub.2. The filtrate was concentrated and purified via flash
chromatography (SiO.sub.2, 25% EtOAc-Hexanes) to yield
(4-{[5-phenylethynyl-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylami-
no]-methyl}-cyclohexyl)-carbamic acid tert-butyl ester.
(4-{[5-Phenylethynyl-2-(2-trifluoromethoxy-benzylamino)pyrimidin-4-ylamin-
o]-methyl}-cyclohexyl)-carbamic acid tert-butyl ester was dissolved
in CH.sub.2Cl.sub.2 (0.5 mL) and TFA (0.5 mL) was added. The
solution was stirred for 2 h and the volatiles were removed and the
crude residue was purified via preparative thin layer
chromatography (SiO.sub.2, 2000 micron thickness, 1:9:90
NH.sub.4OH:CH.sub.3OH:CH.sub.2Cl.sub.2) to afford 49 mg (57% K) of
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-(phenylethynyl)-N.sup.2-[2-(t-
rifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 496.58
[M+1].sup.+.
[0891] The following compounds were prepared following a procedures
similar to that described above: [0892]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-pent-1-yn-1-yl-N.sup.2-[2-(tr-
ifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 462.39
[M+1].sup.+. [0893]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(trifluoromet-
hoxy)benzyl]-5-{[3-(trifluoromethyl)phenyl]ethynyl}pyrimidine-2,4-diamine,
m/z 564.59 [M+1].sup.+. [0894]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-[(2-fluorophenyl)ethynyl]-N.s-
up.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 513.91
[M+1].sup.+. [0895]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(trifluoromethoxy)be-
nzyl]-5-{[2-(trifluoromethyl)phenyl]ethynyl}pyrimidine-2,4-diamine,
m/z 564.62 [M+1].sup.+. [0896]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-[(3-methylphenyl)ethynyl]-N.s-
up.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 510.04
[M+1].sup.+. [0897]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-[(4-methylphenyl)ethynyl])-N.-
sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z
510.03 [M+1].sup.+.
Example 47
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(3-piperidin-1-y-
lbenzyl)pyrimidine-2,4-diamine
##STR00144##
[0899] Palladium (II) acetate (12.3 mg, 0.055 mmol) and
4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (47.7 mg, 0.082
mmol, Xantphos) were combined and the flask was evacuated and
flushed three times with N.sub.2. Degassed PhCH.sub.3 (25 mL) was
added and the solution was stirred for 5 min. 3-Bromobenzonitrile
(1.0 g, 5.49 mmol) and piperidine (561 mg, 6.59 mmol) were added
and the reaction was stirred for another 5 min. Cs.sub.2CO.sub.3
(2.15 g, 6.59 mmol) was added and the flasked was flushed with
N.sub.2 for 1 min then heated to 70.degree. C. for 48 h. The
volatiles were removed and the crude 3-piperidin-1-yl-benzonitrile
(965 mg) was carried further without purification
[0900] 3-Piperidin-1-yl-benzonitrile (965 mg, 5.18 mmol) was
dissolved in MeOH (125 mL). To this solution was added about 1.5 mL
of Raney Nickel suspension in H.sub.2O. The flask was evacuated and
backflushed with N.sub.2. A balloon was filled with H.sub.2 and the
reaction flask was evaluated, filled with H.sub.2, and maintained
under atmospheric pressure. The reaction was stirred vigorously for
2 h, then filtered through a 2 cm thick pad of Celite under a
stream of N.sub.2. The volatiles were removed and the desired
3-piperidin-1-yl-benzylamine (993 mg) was carried further without
purification.
[0901]
{trans-4-[(2-Chloro-5-nitro-pyrimidin-4-ylamino)-methyl]-cyclohexyl-
}-carbamic acid tert-butyl ester (200 mg, 0.52 mmol) was dissolved
in CH.sub.2Cl.sub.2 (10 mL) and Et.sub.3N (0.289 mL, 2.07 mmol) was
added. 3-Piperidin-1-yl-benzylamine (197 mg, 1.04 mmol) was
dissolved in DMA:ETOH (2 mL, 1:1) and added to the reaction
solution. The reaction was stirred at room temperature for 18 h.
The volatiles were removed in vacuo and the crude was purified via
flash chromatography (Sir, 25-50% EtOAc:Hexanes then 5%
MeOH:CH.sub.2Cl.sub.2) to afford 176 mg (63%) of
(4-{[5-nitro-2-(3-piperidin-1-yl-benzylamino)-pyrimidin-4-ylamino]-methyl-
}-cyclohexyl)-carbamic acid tert-butyl ester
[0902]
(4-{[5-Nitro-2-(3-piperidin-1-yl-benzylamino)-pyrimidin-4-ylamino]--
methyl}-cyclohexyl)-carbamic acid tert-butyl ester (176 mg, 0.33
mmol) was dissolved in CH.sub.2Cl.sub.2 (6 mL). To this solution
was added TFA (3 mL) and the reaction was stirred for 1 h. The
volatiles were removed and the crude was re-dissolved in 90:9:1
(CH.sub.2Cl.sub.2:CH.sub.3OH:NH.sub.4OH), then concentrated again
to afford a solid. The resultant solid was purified via flash
chromatography (SiO.sub.2, 10-75%
CH.sub.2Cl.sub.2-(90:9:1-CH.sub.2Cl.sub.2:CH.sub.3OH:NH.sub.4OH))
to afford 131 mg (91%) of the target compound, m/z 440.72
[M+1].sup.+.
[0903] The following compounds were prepared following a procedure
similar to that described above: [0904]
N.sup.4-[(trans-4-aminocyclohexyl)methyl)]-5-nitro-N.sup.2-(3-pyrrolidin--
1-ylbenzyl)pyrimidine-2,4-diamine, m/z 426.68 [M+1].sup.+. [0905]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(3-azepan-1-ylbenzyl)-5-
-nitropyrimidine-2,4-diamine, m/z 454.72 [M+1].sup.+.
Example 48
N-(4-{[2-(3-Bromo-2-methyl-benzylamino)-5-nitro-pyrimidin-4-ylamino]-methy-
l}-cyclohexyl)-2,2,2-trifluoro-acetamide
##STR00145##
[0907] To a solution of the
N.sup.4-(4-Amino-cyclohexylmethyl)-N.sup.2-(3-bromo-2-methyl-benzyl)-5-ni-
tro-pyrimidine-2,4-diamine bis-trifluoroacetate salt (185 mg, 0.273
mmol) in dichloromethane (1.5 mL) was added the methanesulfonyl
chloride (43 .mu.L, 0.546 mmol) and diisopropylethylamine (143
.mu.L, 0.819 mmol) and the solution was stirred for 1 hour. The
reaction mixture was diluted with dichloromethane (2 mL) and
saturated aqueous NaHCO.sub.3 (2 mL) was added. The phases were
separated and the aqueous phase was extracted with dichloromethane
(3 mL). The organic phases were combined, dried over
Na.sub.2SO.sub.4, and concentrated to afford 87.2 mg (59%) of the
target compound, m/z 546.4 [M+1].sup.+
Example 49
3-{[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)amin-
o]methyl}pyridin-2(1H)-one
##STR00146##
[0909]
[trans-4-({2-[(2-Methoxy-pyridin-3-ylethyl)-amino]-5-nitro-pyrimidi-
n-4-ylamino}-methyl)-cyclohexyl]-carbamic acid tert-butyl ester
(100 mg, 0.205 mmol) was dissolved in dioxane (2 mL) and treated
with HCl in dioxane (0.85 mL, 4 mmol/mL, 3.40 mmol). After stirring
overnight, an oil was seen at the bottom of the solution. The
solvent was taken out and the residue dried to give an oily solid.
The crude solid was purified with prep TLC (1:9:90
NH.sub.3:MeOH:DCM) to give the title product (20.5 mg, 0.055 mmol,
26.8%) as a yellow solid, m/z 374.5 (M+1).sup.+
Example 50
N-methyl-N-[1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyr-
imidin-4-yl)amino]methyl}cyclohexyl)pyrrolidin-3-yl]acetamide
##STR00147##
[0911] cis-4-Hydroxy-cyclohexanecarbonitrile was made according to
literature procedures (Noyce, D. S. et al.; J. Org. Chem.; 1969;
34, 1247).
[0912] To a 2 N solution of NH.sub.3 in MeOH (40 mL) in a high
pressure autoclave was added cis-4-hydroxy-cyclohexanecarbonitrile
(6.6 g, 52.7 mmol) and Raney Ni (1.0 g, 17 mmol, 0.3 equiv.) The
mixture was placed under 45 psi H2 and shaken overnight. The
pressure was released and the reaction filtered through a pad of
Celite. The filter pad was washed with EtOAc and the mixture was
concentrated under reduced pressure to provide a yellow oil as
cis-4-aminomethyl-cyclohexanol (6.35 g, 49.1 mmol, 93.2%).
[0913]
(5-Nitro-4-thiocyanato-pyrimidin-2-yl)-(2-trifluoromethoxy-benzyl)--
amine (10 g, 26.9 mmol) was dissolved in DCM (100 ml).
cis-4-aminomethyl-cyclohexanol (3.48 g, 26.9 mmol) and
diisopropylethylamine (10 mL, 53.8 mmol) were added to the solution
and stirred overnight. The reaction mixture was evaporated in-vacuo
and purified by column chromatography (1% MeOH/99% CH2Cl2). The
resulting solid was washed with cold methanol to give a pale yellow
solid as
cis-4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylamino]-m-
ethyl}-cyclohexanol (11.3 g, 25.6 mmol, 95.1%).
[0914]
cis-4-{[5-Nitro-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylam-
ino]-methyl}-cyclohexanol (3.6 g, 8.16 mmol) was dissolved in dry
DCM (150 mL) and cooled to 5.degree. C. Methanesulfonyl chloride
(0.94 ml, 12.2 mmol) and diisopropylethylamine (4.26 ml, 24.4 mmol)
were added sequentially. After 2 h, the reaction was quenched with
cold water (10 mL) and warmed to room temperature. The solution was
partitioned between DCM (100 mL) and water (50 mL). The layers were
separated and the aqueous layer was extracted with DCM (2.times.100
mL). The combined organic layers were washed with brine, dried over
MgSO4, filtered and concentrated in vacuo to yield a yellow solid.
The residue was then chromatographed (silica, 100:1 DCM/MeOH) and
washed with cold methanol to yield a pale yellow solid as
methanesulfonic acid
cis-4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylamino]-m-
ethyl}-cyclohexyl ester (3.90 g, 7.50 mmol, 92%).
[0915] Methanesulfonic acid
4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)pyrimidin-4-ylamino]-methyl-
}-cyclohexyl ester (100 mg, 0.19 mmol) and
N-Methyl-N-pyrrolidin-3-yl-acetamide (137 mg, 0.96 mmol) were
combined with DMA (0.4 mL) and heated at 100 degrees overnight. The
reaction mixture was then dissolved in methanol and purified by
preparatory HPLC. The pure HPLC fractions were lyophilized to give
the title compound (30 mg, 28%), m/z 566.6 [M+1].sup.+.
[0916] The following compounds were prepared following a procedure
similar to that described above: [0917]
N.sup.4-{[trans-4-(1,1-dioxidothiomorpholin-4-yl)cyclohexyl]methyl}-5-nit-
ro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine,
(BI00612608) m/z 559.2 [M+1].sup.+. [0918]
N.sup.4-{[trans-4-(3,3-difluoropyrrolidin-1-yl)cyclohexyl]methyl}-5-nitro-
-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z
531.1 [M+1].sup.+ [0919]
4-trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)-
amino]methyl}cyclohexyl)piperazin-2-one, m/z 524.1 [M+1].sup.+
[0920]
N.sup.4-({trans-4-[4-(methylsulfonyl)piperazin-1-yl]cyclohexyl}methyl)-5--
nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine,
m/z 588.1 [M+1].sup.+ [0921]
N.sup.4-{[trans-4-(4-acetylpiperazin-1-yl)cyclohexyl]methyl}-5-nitro-N.su-
p.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 552.1
[M+1].sup.+ [0922]
N.sup.4-({trans-4-[3-(methylsulfonyl)pyrrolidin-1-yl]cyclohexyl}methyl)-5-
-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine,
m/z 573.1 [M+1].sup.+ [0923]
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)-1,4-diazepan-5-one, m/z 538.1 [M+1].sup.+
[0924]
N-[(3S)-1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimi-
din-4-yl)amino]methyl}cyclohexyl)pyrrolidin-3-yl]acetamide, m/z
552.2 [M+1].sup.+ [0925]
N-[(3R)-1-(trans-4-{[(5-nitro-2-{([2-(trifluoromethoxy)benzyl]amino}pyrim-
idin-4-yl)amino]methyl}cyclohexyl)pyrrolidin-3-yl]acetamide, m/z
552.2 [M+1].sup.+ [0926]
N.sup.4-({trans-4-[(3R)-3-(dimethylamino)pyrrolidin-1-yl]cyclohexyl}methy-
l)-5-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine,
m/z 538.2 [M+1].sup.+ [0927]
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)-L-prolinamide, m/z 538.2 [M+1].sup.+
[0928]
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)-D-prolinamide, m/z 538.1 [M+1].sup.+
[0929]
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)piperidine-3-carboxamide, m/z 552.2
[M+1].sup.+ [0930]
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimid-
in-4-yl)amino]methyl}cyclohexyl)piperidin-3-ol, m/z 525.1
[M+1].sup.+ [0931]
1-(trans-4-{[(5-nitro-2-{[2-trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)piperidin-4-ol, m/z 525.2
[M+1].sup.+ [0932]
5-nitro-N.sup.4-[(trans-4-piperazin-1-ylcyclohexyl)methyl]-N.sup.2-
-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine m/z 510.6
[M+1].sup.+ [0933]
N.sup.4-{[trans-4-(4-methylpiperazin-1-yl)cyclohexyl]methyl}-5-nit-
ro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z
524.6 [M+1].sup.+ [0934] ethyl
4-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)piperazine-1-carboxylate, m/z 582.6
[M+1].sup.+ [0935]
2-[methyl(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}-
pyrimidin-4-yl)amino]methyl}cyclohexyl)amino]ethanol, m/z 499.6
[M+1].sup.+ [0936]
N-(2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin--
4-yl)amino]methyl}cyclohexyl)amino]ethyl)acetamide, m/z 526.6
[M+1].sup.+ [0937]
N-{[trans-4-(1,4-diazepan-1-yl)cyclohexyl]methyl}-5-nitro-N.sup.2--
[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 524.7
[M+1].sup.+ [0938]
N.sup.4-{[trans-4-(4-methyl-1,4-diazepan-1-yl)cyclohexyl]methyl}-5-
-nitro-N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine,
m/z 538.6 [M+1].sup.+ [0939]
N.sup.4-{[trans-4-(4-acetyl-1,4-diazepan-1-yl)cyclohexyl]methyl}-5-nitro--
N.sup.2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z
566.6 [M+1].sup.+ [0940]
N'-{[trans-4-(3-fluoropyrrolidin-1-yl)cyclohexyl]methyl}-5-nitro-N.sup.2--
[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 513.1
[M+1].sup.+ [0941]
N,N-dimethyl-4-trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]a-
mino}pyrimidin-4-yl)amino]methyl}cyclohexyl)piperazine-1-carboxamide,
m/z 581.2 [M+1].sup.+ [0942]
(2R)-1-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]propan-2-ol, m/z 499.6
[M+1].sup.+ [0943]
(2S)-1-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}p-
yrimidin-4-yl)amino]methyl}cyclohexyl)amino]propan-2-ol, m/z 499.6
[M+1].sup.+ [0944]
3-[(trans-4-{[5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)amino]propan-1-ol, m/z 499.2 [M+1].sup.+
[0945]
N.sup.4-({trans-4-[(3-aminopropyl)amino]cyclohexyl}methyl)-5-nitro-N.sup.-
2-[2-(trifluoromethoxy)benzyl]pyrimidine-2,4-diamine, m/z 498.1
[M+1].sup.+ [0946]
(2R-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-
-4-yl)amino]methyl}cyclohexyl)amino]propan-1-ol, m/z 499.1
[M+1].sup.+ [0947]
(2S)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}p-
yrimidin-4-yl)amino]methyl}cyclohexyl)amino]propan-1-ol, m/z 499.1
[M+1].sup.+ [0948]
(2R)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]-3-phenylpropan-1-ol, m/z
575.6[M+1].sup.+ [0949]
(2S)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]-3-phenylpropan-1-ol, m/z
575.6 [M+1].sup.+ [0950]
N.alpha.-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimid-
in-4-yl)amino]methyl}cyclohexyl)-L-phenylalaninamide, m/z 588.6
[M+1].sup.+ [0951]
N.alpha.-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimid-
in-4-yl)amino]methyl}cyclohexyl)-D-phenylalaninamide, m/z 588.6
[M+1].sup.+ [0952]
N,N-dimethyl-1-trans-4-{[(5-nitro-2-{[2-trifluoromethoxy)benzyl]amino}pyr-
imidin-4-yl)amino]methyl}cyclohexyl)-L-prolinamide, m/z 566.1
[M+1].sup.+
Example 51
N.sup.4-{[trans-(1H-Imidazol-1-yl)cyclohexyl]methyl}-5-nitro-N.sup.2-[2-(t-
rifluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00148##
[0954] A stirred mixture of (4-amino-cyclohexylmethyl)-carbamic
acid benzyl ester (285 mg, 1.09 mmol) in dioxane (1 mL) and water
(2.5 mL) was acidified (pH 1-2) with a few drops of phosphoric
acid. Paraformaldehyde (34 mg) and 40% aqueous glyoxal solution
(150 .mu.L, 1.09 mmol) were added before heating at 80.degree. C.
for 20 min. NH.sub.4Cl (58 mg, 1.09 mmol) was then added and the
mixture heated at 100.degree. C. for a further 3 h. After cooling
to room temperature 2 N NaOH was added until the reaction mixture
attained pH 12. The aqueous phase was extracted with EtOAc
(2.times.20 mL) and the combined extracts dried over
Na.sub.2SO.sub.4 before concentrating in vacuo. The residue was
purified by column chromatography using a 12 g ISCO combi-flash
cartridge (silica gel, 95:4.75:0.25 DCM/MeOH/NH.sub.4OH to obtain
(4-imidazol-1-yl-cyclohexylmethyl)-carbamic acid benzyl ester (60
mg, 17%) as a colorless oil that solidified on standin.
[0955] To a solution of (4-imidazol-1-yl-cyclohexylmethyl)-carbamic
acid benzyl ester (57 mg, 0.18 mmol) in EtOH (4 mL) under N.sub.2
was added 100% Pd/C (200 mg). The reaction flask was flushed with
H2 via balloon and stirred at room temperature for 4 h whereupon
TLC analysis (95:4.75:0.25 DCM/MeOH/NH.sub.4OH) showed complete
consumption of starting material. The reaction mixture was filtered
through a plug of celite and the filtrate concentrated in vacuo to
afford crude (4-imidazol-1-yl-cyclohexyl)-methylamine (32 mg, 95%).
This material was used directly in the next reaction.
[0956] To a solution of (4-imidazol-1-yl-cyclohexyl)-methylamine
(32 mg, 0.18 mmol) and Et.sub.3N (76 .mu.L, 0.54 mmol) in EtOH (1
mL) was added
(5-Nitro-4-thiocyanato-pyrimidin-2-yl)-(2-trifluoromethoxy-benzyl)-amine
(71 mg, 0.19 mmol). The mixture was stirred at room temperature for
14 h before concentrating in vacuo. The residue was partially
purified by column chromatography using a 12 g ISCO combi-flash
cartridge (silica gel, 95:4.75:0.25 DCM/MeOH/NH.sub.4OH) then
re-purified by semi-preparative HPLC. The isolated TFA salt was
converted to the free base by partitioning between sat. NaHCO.sub.3
and EtOAc. Concentration of the organic layer in vacuo afforded 29
mg of the title compound as an off-white solid, m/z 492
[M+1].sup.+
Example 52
N.sup.2-[3-(aminomethyl)benzyl]-5-nitro-N.sup.4-[(trans-4-pyrrolidin-1-ylc-
yclohexyl)methyl]pyrimidine-2,4-diamine
##STR00149##
[0958] To a solution of m-xylylenediamine (5.00 g, 36.70 mmol) in
MeOH (65 mL) was added a solution of Boc.sub.2O (2.00 g, 9.16 mmol)
in MeOH (35 mL) dropwise over 45 min. The reaction mixture was
allowed to stir at room temperature overnight before concentrating
in vacuo. The residue was purified by column chromatography (silica
gel, DCM to 80:19:1 DCM/MeOH/NH.sub.4OH) to afford
(3-aminomethyl-benzyl)-carbamic acid tert-butyl ester (1.43 g,
67%).
[0959] To a solution of 2-chloro-5-nitro-4-thiocyanato-pyrimidine
(1.31 g, 6.05 mmol) in EtOH (10 mL) at 0.degree. C. was added a
solution of (3-aminomethyl-benzyl)-carbamic acid tert-butyl ester
(1.43 g, 6.05 mmol) dropwise over 45 min. The reaction was allowed
to stir at room temperature overnight and the resulting precipitate
filtered and washed with EtOH. The crude material was purified by
column chromatography (silica gel, 10:90 to 12:88 EtOAc/hexane) to
afford
{3-[(5-nitro-4-thiocyanato-pyrimidin-2-ylamino)-methyl]-benzyl}-carbamic
acid tert-butyl ester (1.43 g, 56%) as a yellow solid.
[0960] To a solution of (4-pyrrolidin-1-yl-cyclohexyl)-methylamine
(32 mg, 0.18 mmol) and Et.sub.3N (26 .mu.L, 0.18 mmol) in EtOH (2
mL) was added
{3-[(5-nitro-4-thiocyanato-pyrimidin-2-ylamino)-methyl]-benzyl}-carbamic
acid tert-butyl ester (74 mg, 0.18 mmol). The reaction mixture was
stirred at room temperature overnight before concentrating in
vacuo. The residue was purified by chromatography using an ISCO
combi-flash cartridge (silica gel, 50:50 to 0:100 DCM/80:18:2
DCM/MeOH/NH.sub.4OH) to afford impure
[3-({5-nitro-4-[(4-pyrrolidin-1-yl-cyclohexylmethyl)-amino]-pyrimidin-2-y-
lamino}-methyl)-benzyl]-carbamic acid tert-butyl ester (132 mg),
m/z 540 (M+1).sup.+
[0961] To a solution of
[3-({5-nitro-4-[(4-pyrrolidin-1-yl-cyclohexylmethyl)amino]-pyrimidin-2-yl-
amino}-methyl)-benzyl]-carbamic acid tert-butyl ester (132 mg, 0.25
mmol) in DCM (1 mL) was added TFA (2 mL). After 20 min the reaction
mixture was concentrated in vacuo and the resulting oil taken up in
EtOAc and the solution washed with sat. NaHCO.sub.3. The aqueous
layer was then extracted with 90:10 CHCl.sub.3/i-PrOH. The combined
organic layers were concentrated in vacuo and the residue purified
by column chromatography (silica gel, 96:4 to 93:7 DCM/10%
NH.sub.4OH in MeOH) to afford the title compound (25 mg), m/z 440
(M+1).sup.+.
Preparation of (4-pyrrolidin-1-yl-cyclohexyl)-methylamine
##STR00150##
[0963] To a stirred slurry of (4-aminomethyl-cyclohexyl)-carbamic
acid tert-butyl ester (500 mg, 2.19 mmol) and
N,N-diisopropylethylamine (382 .mu.L, 2.63 mmol) in DCM (8 mL) at
0.degree. C. under N.sub.2 was added benzylchloroformate (311
.mu.L, 2.41 mmol) dropwise via syringe. After 30 min the reaction
was allowed to warm to room temperature and stirred for a further 2
h before diluting with DCM (50 mL). The solution was washed
successively with 1 N HCl, sat. NaHCO.sub.3 and brine before drying
over Na.sub.2SO.sub.4. Residual chloroformate was removed by
chromatography using a 12 g ISCO combi-flash cartridge (silica gel,
90:10 to 50:50 hexane/EtOAc) to afford the di-protected
intermediate which was treated directly with DCM (2 mL) and TFA (2
mL). After 1 h the volatiles were removed in vacuo and the residue
partitioned between sat. NaHCO.sub.3 and EtOAc. Concentration of
the organic layer in vacuo afforded
(4-amino-cyclohexylmethyl)-carbamic acid benzyl ester (517 mg, 90%)
as a white solid.
[0964] To a solution of (4-amino-cyclohexylmethyl)-carbamic acid
benzyl ester (350 mg, 1.34 mmol) in DMF (5 mL) was added
K.sub.2CO.sub.3 (924 mg, 6.70 mmol) and 1,4-dibromobutane (160 mL,
1.34 mmol). The reaction mixture was stirred at room temperature
for 14 h before diluting with EtOAc (100 mL). The reaction solution
was washed with water (2.times.100 mL) and brine before
concentrating in vacuo. The residue was purified by chromatography
using a 12 g ISCO combiflash cartridge (silica gel, 95:4.75:0.25
DCM/MeOH/NH.sub.4OH) to afford
(4-pyrrolidin-1-yl-cyclohexylmethyl)-carbamic acid benzyl ester
(351 mg, 83%) as an off-white solid.
[0965] To a solution of
(4-pyrrolidin-1-yl-cyclohexylmethyl)-carbamic acid benzyl ester
(292 mg, 0.92 mmol) in MeOH (5 mL) was added ammonium formate (0.58
g, 9.20 mmol). The reaction flask was flushed with N.sub.2 prior to
addition of 10% Pd/C (300 mg). The mixture was heated at 65.degree.
C. for 2 h whereupon TLC analysis (silica gel, 95:4.75:0.25
DCM/MeOH/NH.sub.4OH) showed complete consumption of 57. After
cooling to room temperature the reaction mixture was filtered
through a plug of celite in order to remove the Pd catalyst. The
filtrate contained (4-pyrrolidin-1-yl-cyclohexyl)-methylamine which
was used without further purification in the next step.
Example 53
3-{[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-y-
l)amino]methyl}cyclohexyl)amino]methyl}benzoic acid
##STR00151##
[0967] To a mixture of
N.sup.4-(4-amino-cyclohexylmethyl)-5-nitro-N.sup.2-(2-trifluoromethoxy-be-
nzyl)-pyrimidine-2,4-diamine (100 mg, 0.23 mmol) and
3-formyl-benzoic acid methyl ester (41 mg, 0.25 mmol) in DCM (1 mL)
was added sodium triacetoxyborohydride (58 mg, 0.27 mmol). The
mixture was stirred at room temperature for 18 h then quenched with
excess water. The mixture was diluted with DCM and the organic
layer separated and dried over Na.sub.2SO.sub.4. The solvent was
removed in vacuo and the residue purified by column chromatography
using an ISCO combi-flash cartridge (silica gel, DCM/MeOH) to
afford
3-[(4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylamino]-m-
ethyl}-cyclohexylamino)-methyl]-benzoic acid methyl ester (76 mg,
57%), m/z 589 (M+1).sup.+.
[0968] A mixture of
3-[(4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylamino]-m-
ethyl}-cyclohexylamino)-methyl]-benzoic acid methyl ester (75 mg,
0.13 mmol), LiOH (30 mg, 1.29 mmol) and water (3 drops) in MeOH (2
mL) was heated at 50.degree. C. for 18 h. The reaction mixture was
concentrated in vacuo then taken up in water and EtOAc. Dilute HCl
was added until the aqueous phase reached pH 6. The solid that
precipitated was isolated by filtration to afford the target
compound (20 mg, 30%) as a white solid, m/z 575 (M+1).sup.+.
[0969] The following compound was prepared following a procedure
similar to that described above: [0970]
4-{[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4--
yl)amino]methyl}cyclohexyl)amino]methyl}benzoic acid, m/z 575
(M+1).sup.+.
Example 54
N-(trans-4-{[(5-nitro-2-{[2-trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)a-
mino]methyl}cyclohexyl)glycine
##STR00152##
[0972] To a solution of
N.sup.4-(4-amino-cyclohexylmethyl)-5-nitro-N.sup.2-(2-trifluoromethoxy-be-
nzyl)-pyrimidine-2,4-diamine (88 mg, 0.20 mmol) in 2:1 DMF/DMSO
(0.6 mL) was added N,N-diisopropylethylamine (35 .mu.L, 0.30 mmol)
followed by tert-butyl bromoacetate (22 mL, 0.15 mmol). The mixture
was stirred at room temperature overnight then partitioned with
EtOAc (15 mL) and water (5 mL). The layers were separated and the
organics washed with water (2.times.5 mL) and brine. After
concentration in vacuo, the residue was purified by column
chromatography (silica gel, 97:3 DCM/MeOH) to afford
(4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylamino]-meth-
yl}-cyclohexylamino)acetic acid tert-butyl ester (51 mg, 61%) as a
colorless oil, m/z 555 [M+1].sup.+.
[0973] To a solution of
(4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylamino]-meth-
yl}-cyclohexylamino)-acetic acid tert-butyl ester (60 mg, 0.108
mmol) in DCM (1 mL) was added TFA (1 mL). The solution was allowed
to stand at room temperature for 16 h before the solvent was
removed in vacuo. The residue was taken up in water (1 mL) and 2 N
NaOH added until the mixture reached pH 5. The mixture was diluted
with water (3 mL) and filtered. The filtered solid was washed with
water and Et.sub.2O before drying in vacuo to afford the target
compound (46 mg, 85%) as a white solid, m/z 499 [M+1].sup.+.
[0974] The following compound was prepared following similar
procedures as described above: [0975]
N-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}-cyclohexyl)-beta-alanine, m/z 513 (M+1).sup.+.
Example 55
4-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)amino]butanoic acid
##STR00153##
[0977] To a solution of
N.sup.4-(4-amino-cyclohexylmethyl)-5-nitro-N.sup.2-(2-trifluoromethoxy-be-
nzyl)-pyrimidine-2,4-diamine (80 mg, 0.182 mmol) in 2:1 DMF/DMSO
(0.6 mL) was added N,N-diisopropylethylamine (32 .mu.L, 0.272 mmol)
followed by methyl-4-iodobutyrate (19 .mu.L, 0.136 mmol). The
mixture was stirred at room temperature overnight then partitioned
with EtOAc (15 mL) and water (10 mL). The layers were separated and
the organics dried over Na.sub.2SO.sub.4. After concentration in
vacuo, the residue was purified by column chromatography using a 12
g ISCO combi-flash cartridge (silica gel, 100:0 to 90:10 DCM/MeOH)
to afford
4-(4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)-pyrimidin-4-ylamino]-me-
thyl}-cyclohexylamino)-butyric acid methyl ester (39 mg, 53%) as a
white solid: m/z 541 [M+1].sup.+.
[0978] To a solution of
4-(4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)pyrimidin-4-ylamino]-met-
hyl}-cyclohexylamino)-butyric acid methyl ester (39 mg, 0.072 mmol)
in 6:1 MeOH/water (0.7 mL) was added LiOH (9 mg, 0.375 mmol) and
the mixture stirred at room temperature overnight. The resulting
suspension was diluted with MeOH (1 mL) and stirred at room
temperature for a further 2 h. The reaction mixture was
concentrated in vacuo and the remaining aqueous residue treated
with 2 N HCl until the mixture reached pH 5-6. The resulting solid
was filtered and washed with water. The material was subjected to
semi-preparative to separate the title compound [7 mg, 13%, m/z 527
(M+1).sup.+] and
1-(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl-
)amino]methyl}cyclohexyl)pyrrolidin-2-one [15 mg, 41%, m/z 509
(M+1).sup.+].
Example 56
4-{[(4-{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)amin-
o]methyl}-5-chloropyridin-2(1H)one
##STR00154##
[0980] A mixture of 3,6-dichloro-4-cyanopyridine (prepared
according to WO 9633975) (3.40 g, 19.65 mmol) and PtO.sub.2 (0.40
g) in 1:1 Ac.sub.2O/AcOH (60 mL) was shaken in a Parr hydrogenation
apparatus at 30 psi H.sub.2 for 6 h. The reaction mixture was
filtered through a pad a celite and the filtered solids washed with
EtOAc. The combined filtrate and washings were concentrated in
vacuo and the resulting white solid purified by column
chromatography on a 120 g ISCO combi-flash cartridge (silica gel,
20:80 to 100:0 EtOAc/hexane) to afford
N-(2,5-Dichloro-pyridin-4-ylmethyl)-acetamide (1.73 g, 40%) as a
white solid.
[0981] Compound N-(2,5-dichloro-pyridinylmethyl)-acetamide (250 mg,
1.14 mmol) was added to 33% H.sub.2SO.sub.4 (3 mL) and the mixture
heated in a CEM microwave reactor at 200.degree. C. for 30 min.
After cooling to room temperature, the reaction mixture was diluted
with water (20 mL) and basified with excess concentrated
NH.sub.4OH. The reaction mixture was then frozen using a dry
ice/acetone bath and evaporated to dryness in vacuo overnight in a
freeze-dry apparatus. The solids were purified by column
chromatography using a 12 g ISCO combi-flash column and dry-loading
cartridge (silica gel, 100:0 to 50:50 DCM:DCM/MeOH/NH.sub.4OH
6:3:1) to afford
N-(5-chloro-2-oxo-1,2-dihydro-pyridin-4-ylmethyl)acetamide (246 mg,
quant.) as an off-white solid. This material was directly in the
next step.
[0982] A mixture of
N-(5-chloro-2-oxo-1,2-dihydro-pyridin-4-ylmethyl)-acetamide (100
mg, 0.62 mmol),
(4-[(2-chloro-5-nitro-pyrimidin-4-ylamino)methyl]-cyclohexyl)-carb-
amic acid tert-butyl ester (146 mg, 0.38 mmol) and
diisopropylethylamine (188 .mu.L, 1.14 mmol) in i-PrOH (2 mL) was
heated in a CEM microwave reactor at 120.degree. C. for 30 min.
After cooling the reaction mixture was concentrated in vacuo and
the residue treated with TFA (10 mL). After 30 min the solution was
concentrated in vacuo and basified with excess 6:3:1
DCM/MeOH/NH.sub.4OH. Concentration in vacuo afforded a residue
which was purified by column chromatography using a 12 g ISCO
combi-flash cartridge (silica gel, 50:50 to 0:100 DCM/(6:3:1
DCM/MeOH/NH.sub.4OH). The material obtained after chromatography
was then suspended in EtOAc (20 mL) and the flask placed in a
sonicator bath for 5 min. The insoluble material was removed by
filtration and the filtrate concentrated in vacuo to afford the
target compound (148 mg, 96%) as a white solid, m/z 408
[M+1].sup.+.
[0983] The following compound was prepared following similar
procedures as described above: [0984] 4-{[(4
{[(trans-4-aminocyclohexyl)methyl]amino}-5-nitropyrimidin-2-yl)amino]meth-
yl}pyridin-2(1H)-one, m/z 374 [M+1].sup.+.
Example 57
(1S)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-
-4-yl)amino]methyl}cyclohexyl)amino]-1-phenylethanol and
(2R)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]-2-phenylethanol
##STR00155##
[0986] A solution of
N.sup.4-trans-(4-amino-cyclohexylmethyl)-5-nitro-N.sup.2-(2-trifluorometh-
oxy-benzyl)-pyrimidine-2,4-diamine (400 mg, 0.91 mmol) and
(S)-styrene oxide (109 mg, 0.91 mmol) in EtOH (5 mL) was stirred at
80.degree. C. for 72 h. After concentration in vacuo, the residue
was purified by preparative TLC (silica gel, 90:10 DCM/MeOH) to
afford
(1S)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]-1-phenylethanol, [52 mg, m/z
562 (M+1).sup.+] and
(2R)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}-pyrimid-
in-4-yl)amino]methyl}cyclohexyl)amino]-2-phenylethanol [6 mg, m/z
561 (M+1).sup.+] as off-white solids.
[0987] The following compounds were prepared following similar
procedures as described above: [0988]
(1R)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidi-
n-4-yl)amino]methyl}cyclohexyl)amino]-1-phenylethanol, m/z
562[M+1].sup.+. [0989]
(2S)-2-[(trans-4-{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}p-
yrimidin-4-yl)amino]methyl}cyclohexyl)amino]-2-phenylethanol, m/z
562[M+1].sup.+.
Example 58
N.sup.4-[(trans-4-amino-4-methylcyclohexyl)methyl]-5-nitro-N.sup.2-[2-(tri-
fluoromethoxy)benzyl]pyrimidine-2,4-diamine
##STR00156##
[0991] To a stirred slurry of
trans-4-aminomethyl-cyclohexanecarboxylic acid (5.00 g, 31.8 mmol)
and K.sub.2CO.sub.3 (4.40 g, 127.2 mmol) in MeOH (100 mL) cooled to
0.degree. C. was added benzyl bromide (7.94 mL, 66.8 mmol). The
reaction mixture was allowed to warm to room temperature and
stirred overnight. Most of the volatiles were removed in vacuo
before addition of water (100 mL). The aqueous mixture was
extracted with Et.sub.2O (2.times.200 mL) before adjusting to pH
5-6 by addition of 2 N HCl. The aqueous phase was extracted with
DCM (3.times.200 mL) and the combined DCM extracts dried over
Na.sub.2SO.sub.4 before concentrating in vacuo to afford
trans-4-[(dibenzylamino)-methyl]-cyclohexanecarboxylic acid (3.22
g, 43%) as an off-white solid, m/z 338 [M+1].sup.+.
[0992] To a solution of diisopropylamine (0.95 g, 9.4 mmol) in THF
(40 mL) under N2 at -78.degree. C. was added n-BuLi (3.75 mL of a
2.5 M solution in hexanes, 9.38 mmol) dropwise via syringe. After
20 min a solution of
trans-4-[(dibenzylamino)-methyl]-cyclohexanecarboxylic acid (1.58
g, 4.7 mmol) in THF (10 mL) was added dropwise. The reaction
mixture was stirred at -78.degree. C. for a further 20 min before
it was allowed to warm to room temperature and stirred for a
further 3 h. The reaction mixture was cooled to -78.degree. C. and
methyl iodide (0.32 mL, 5.2 mmol) added dropwise. The reaction was
then allowed to warm slowly to room temperature and stirred
overnight before quenching with water (10 mL). The mixture was
treated with 6 N HCl until it reached pH 6 and was then
concentrated in vacuo to remove most of the THF. The aqueous phase
was extracted with EtOAc (2.times.60 mL) and the combined extracts
washed with brine, dried (Na.sub.2SO.sub.4) and concentrated in
vacuo. The residue was purified by column chromatography (silica
gel, 1:1 hexane/Et.sub.2O) to afford
trans-4-[(dibenzylamino)-methyl]-1-methyl-cyclohexanecarboxylic
acid (375 mg, 23%), m/z 352 (M+1).sup.+.
[0993] To a solution of
trans-4-[(dibenzylamino)-methyl]-1-methyl-cyclohexanecarboxylic
acid (375 mg, 1.07 mmol) and Et.sub.3N (317 .mu.L, 2.28 mmol) in
toluene (3.6 mL) under N.sub.2 was added diphenylphosphoryl azide
(250 .mu.L, 1.16 mmol). The reaction was stirred at room
temperature for 1 h before heating at 100.degree. C. for 4 h. The
reaction mixture was concentrated to about half volume then a
solution of Boc.sub.2O (0.70 g, 3.20 mmol) in THF (4 mL) added
followed by Et.sub.3N (0.60 mL, 4.30 mmol) and water (0.2 mL).
After stirring at room temperature for 2 h, little or no reaction
was observed and the intermediate isocyanate remained unchanged.
The solvent was evaporated in vacuo before the residue was
dissolved in 1:1 THF/6 N HCl (4 mL) and stirred at room temperature
overnight. NaOH pellets were added until the reaction mixture
reached pH 11 and Boc.sub.2O (0.70 g, 3.20 mmol) added. After
stirring at room temperature for 3 h the reaction mixture was
extracted with EtOAc (2.times.30 mL) and the combined extracts
dried over Na.sub.2SO.sub.4. After concentration in vacuo the
residue was purified by column chromatography using a 12 g ISCO
combi-flash cartridge (silica gel, 90:10 to 50:50 hexane/EtOAc) to
afford
trans-{4-[(dibenzylamino)-methyl]-1-methyl-cyclohexyl}-carbamic
acid tert-butyl ester (221 mg, 50%) as a colorless oil, m/z 423
[M+1].sup.+.
[0994] To a mixture of
trans-{4-[(dibenzylamino)-methyl]-1-methyl-cyclohexyl}-carbamic
acid tert-butyl ester (221 mg, 0.52 mmol) and ammonium formate (330
mg, 5.23 mmol) in MeOH (5 mL) under N.sub.2 was added 10% palladium
on charcoal (300 mg). The reaction was heated at 60.degree. C. for
1 h and allowed to cool to room temperature before filtering
through celite. Concentration in vacuo afforded
trans-(4-aminomethyl-1-methyl-cyclohexyl)-carbamic acid tert-butyl
ester (126 mg, 100%) as a colorless oil, m/z 243 [M+1].sup.+.
[0995] A solution of
trans-{4-[(dibenzylamino)-methyl]-1-methyl-cyclohexyl}-carbamic
acid tert-butyl ester (62 mg, 0.26 mmol) was added to
(5-nitro-4-thiocyanato-pyrimidin-2-yl)-(2-trifluoromethoxy-benzyl)-amine
(95 mg, 0.26 mmol) followed by Et.sub.3N (143 .mu.L, 1.03 mmol).
The mixture was stirred at room temperature for 3 h then
concentrated in vacuo. The residue was purified by column
chromatography using a 12 g ISCO combi-flash cartridge (silica gel,
90:10 to 70:30 hexane/EtOAc) to afford
trans-(1-methyl-4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)-pyr-
imidin-4-ylamino]-methyl}-cyclohexyl)-carbamic acid tert-butyl
ester (111 mg, 79%) as an off-white solid, m/z 555 [M+1].sup.+.
[0996] To a solution of
trans-(1-methyl-4-{[5-nitro-2-(2-trifluoromethoxy-benzylamino)-pyrimidin--
4-ylamino]-methyl}-cyclohexyl)-carbamic acid tert-butyl ester (110
mg, 0.20 mmol) in DCM (2 mL) at room temperature was added TFA
(2-mL). After 1 h the reaction mixture was concentrated in vacuo
and treated with sat. NaHCO.sub.3 (5 mL) before extracting with
EtOAc (2.times.25 mL). The combined organic layers were dried
(Na.sub.2SO.sub.4) and concentrated in vacuo to afford the title
compound (83 mg, 92%) as an off-white solid: m/z 455
[M+1].sup.+.
Example 59
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-(1,2-diphenylethyl)-5-ni-
tropyrimidine-2,4-diamine
##STR00157##
[0998]
trans-{4-[(2-Chloro-5-nitro-pyrimidin-4-ylamino)-methyl]-cyclohexyl-
}-carbamic acid tert-butyl ester (100 mg, 0.26 mmol) was dissolved
in DCM (5 mL). 1,2-Diphenylethylamine (154 mg, 0.78 mmol) was added
in one portion. After 18 h, the solution was partitioned between
DCM (15 mL) and 2% AcOH (aq; 10 mL). The layers were separated and
the aqueous layer was extracted with DCM (20 mL). The combined
organic fraction were washed with saturated sodium carbonate (10
mL) and water, dried over MgSO.sub.4, filtered and concentrated to
yield a pale yellow solid. Chromatography (silica, 10%-50%
Hex/EtOAc) yielded 110 mg of
(4-{[2-(1,2-diphenyl-ethylamino)-5-nitro-pyrimidin-4-ylamino]-methyl}-cyc-
lohexyl)-carbamic acid tert-butyl ester as a pale yellow solid, m/z
547.4 [M+1].sup.+.
[0999]
(4-{[2-(1,2-Diphenyl-ethylamino)-5-nitro-pyrimidin-4-ylamino]-methy-
l}-cyclohexyl)-carbamic acid tert-butyl ester (105 mg, 0.19 mmol)
was dissolved in dioxane (5 mL). An HCl solution (2 mL of a 4M
solution) was added. After 18 h, an oil separated from the solvent.
The solvent was decanted and the oil was dissolved in water and
lyophilized yielding 57 mg of the title compound as a pale yellow
solid, m/z 447.3 [M+1].sup.+.
[1000] The following compound was prepared following similar
procedures as described above: [1001]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-5-nitro-N.sup.2-(2-piperidin-1--
ylbenzyl)pyrimidine-2,4-diamine, m/z 440.6 [M+1].sup.+.
Example 60
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[2-(methylsulfonyl)benzy-
l]-5-nitropyrimidine-2,4-diamine
##STR00158##
[1003]
{4-[(2-Chloro-5-nitro-pyrimidin-4-ylamino)-methyl]-cyclohexyl}-carb-
amic acid tert-butyl ester (100 mg, 0.26 mmol) was dissolved in DMF
(3 mL). 2-Methanesulfonyl-benzylamine hydrochloride (115 mg, 0.52
mmol) and diisopropylethylamine (0.14 mL, 0.78 mmol) were added
sequentially. After 24 h, the reaction was partitioned between
water (10 mL) and DCM (10 mL). The organic layer was washed with 5%
AcOH (aq), saturated Na.sub.2CO.sub.3 and water, dried over
MgSO.sub.4, filtered and concentrated. Chromatography (silica,
0-15% MeOH in DCM; Rf-0.3 10% MeOH in DCM) yielded 96 mg of
(4-{[2-(2-methanesulfonyl-benzylamino)-5-nitro-pyrimidin-4-ylamino]-methy-
l}-cyclohexyl)-carbamic acid tert-butyl ester as a pale yellow
solid, m/z 535.7 [M+1].sup.+.
[1004]
(4-{[2-(2-Methanesulfonyl-benzylamino)-5-nitro-pyrimidin-4-ylamino]-
-methyl}-cyclohexyl)-carbamic acid tert-butyl ester (96 mg, 0.18
mmol) was dissolved in warn dioxane (5 mL). HCl in dioxane (4M, 1.5
mL, 6.0 mmol) was added. After 18 h, the heterogeneous mixture was
decanted and the solid was washed with dioxane and dried in-vacuo
to yield 76 mg of the title compound as a white solid, m/z 435.5
[M+1]*.
Preparation of 2-Methanesulfonyl-benzylamine intermediate
##STR00159##
[1006] 2-(Methylthio)benzyl amine (3 g, 19.6 mmol) was dissolved in
DCM (100 mL). Di-tert-butyldicarbonate (4.27 g, 19.6 mmol) was
added portionwise. After 8 h, the reaction was concentrated.
Chromatography (silica, Hex/EtOAc 2%-15%) yielded 4.5 g of a
(2-methylsulfanyl-benzyl)-carbamic acid tert-butyl ester as a
colorless oil, m/z 254.4 [M+1].sup.+.
[1007] MCPBA (60%, 5.82 g, 20.2 mmol) was added to a solution of
(2-methylsulfanyl-benzyl)-carbamic acid tert-butyl ester (2.5 g,
9.87 mmol) in DCM (100 mL) which was pre-cooled to 0 degrees C.
After 5 h, 100 ml of a 10% aqueous solution of sodium sulfite
(Na.sub.2SO3) was added to the reaction and the organic layers were
collected, washed with saturated aqueous sodium carbonate (50 mL),
dried over MgSO4, filtered and concentrated in-vacuo.
Chromatography (silica, 0-15% MeOH in DCM) yielded 1.885 g of
(2-methanesulfonyl-benzyl)-carbamic acid tert-butyl ester as an
off-white solid.
[1008] (2-Methanesulfonyl-benzyl)-carbamic acid tert-butyl ester
(1.55 g, 5.43 mmol) was dissolved in warm dioxane (50 mL) and
cooled to room temperature. HCl solution (4M solution in dioxane,
20 ml, 80 mmol) was added. After 20 h, a white precipitate had
formed. The reaction was diluted with CHCl.sub.3 (50 mL) and
filtered. The white solid was washed with CHCl.sub.3 and dried
in-vacuo to yield 1.075 g of 2-methanesulfonyl-benzylamine
hydrochloride as an off white solid, m/z 186.6 (M+1).sup.+.
[1009] The following compound was prepared following similar
procedures as described above: [1010]
N.sup.4-[(trans-4-aminocyclohexyl)methyl]-N.sup.2-[5-chloro-2-(methylsulf-
onyl)benzyl]-5-nitropyrimidine-2,4-diamine, m/z 469.6
(M+1).sup.+.
[1011] The following intermediate was prepared following similar
procedures as described above for 2-methanesulfonyl-benzylamine:
[1012] 5-Chloro-2-methanesulfonyl-benzylamine, m/z 220.5
(M+1).sup.+.
Preparation of 5-chloro-2-methylsulfanyl-benzylamine
intermediate
##STR00160##
[1014] 5-Chloro-2-fluorobenzonitrile (9.0 g, 57.9 mmol) was
dissolved in DMF (200 mL). Sodium methanethiolate (4.86 g, 69.4
mmol) was added in one portion and the reaction was heated to 80
degrees C. After 72 h, the reaction was cooled to room temperature
and partitioned between EtOAc (250 mL) and H.sub.2O (250 mL). The
layers were separated and the aqueous layer was extracted with
EtOAc (150 mL). The combined organic layers were washed with water
(3.times.250 mL) and brine (50 mL), dried over MgSO.sub.4, filtered
and concentrated to yield 11 g of a yellow solid. Recrystallization
from EtOH yielded 7.5 g (two batches) of
5-chloro-2-methylsulfanyl-benzonitrile as pale yellow crystals.
[1015] 5-Chloro-2-methylsulfanyl-benzonitrile (3.9 g, 21.2 mmol)
was dissolved in THF (42 mL) and cooled to 0 degrees C. Borane-THF
complex (50 mL of a 1M THF solution) was added dropwise. After 10
minutes at 0 degrees C. the reaction was warmed to 65 degrees C.
for 4 h. Methanol (approximately 30 mL) was added to the reaction
at 0 degrees C. The reaction was warmed to room temperature and
stirred for 10 minutes. The reaction solution was concentrated
in-vacuo to yield a thick oil which was dissolved in 50% THF/MeOH
(30 mL). 4M HCl (30 mL) was added slowly. After 10 minutes, the
solution was partitioned between water (10 mL) and DCM (20 mL). The
aqueous layer was extracted with DCM (30 mL) and the organic layers
were discarded. The aqueous layer was neutralized to pH9 with 5M
NaOH. The solution was extracted with DCM (3.times.70 mL). The
combined organic layers were washed with brine, dried over
MgSO.sub.4, filtered and concentrated to yield 2.9 g of
5-chloro-2-methylsulfanyl-benzylamine as a pale yellow oil, .sup.1H
NMR 400 MHz (CDCl.sub.3) .delta. 7.33 (d, 1H, J=2.4 Hz), 7.22 (dd,
1H, J=2.4, 8.4 Hz), 7.13 (d, 1H, J=8.4 Hz), 3.89 (s, 2H), 2.48 (s,
3H).
Example 61
cis-2-amino-trans-5{[(5-nitro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimid-
in-4-yl)amino]methyl}cyclohexanol
##STR00161##
[1017] trans-4-Azido-cis-3-hydroxy-cyclohexanecarboxylic acid
methyl ester (8.66 g, 43.5 mmol) was combined with PPh.sub.3 (13.7
g, 52.2 mmol) and benzoic acid (6.4 g, 52.2 mmol) and dissolved in
THF (150 mL). The solution was cooled to 0 degrees C. and DEAD (8.2
mL, 52.2 mmol) was added dropwise over 20 minutes. The reaction was
stirred at 0 degrees C. for 24 h. The reaction was then warmed to
room temperature. After 6 h at room temperature, the reaction was
partitioned between saturated NaHCO.sub.3 (100 mL) and Et.sub.2O
(300 mL). The layers were separated and the aqueous layer was
extracted with Et.sub.2O (2.times.250 mL). The combined organic
layers were washed with brine, dried over MgSO.sub.4, filtered and
concentrated to yield a colorless oil. The oil was dissolved in
Et.sub.2O (approx. 200 mL) and the heterogeneous solution was
filtered and the white solid was washed with Et.sub.2O (75 mL). The
filtrate was concentrated in vacuo. The oil was then stirred with
5% EtOAc in Hex (500 mL) over 30 minutes. The heterogeneous
solution was filtered and the filtrate was concentrated in vacuo.
Chromatography (Combiflash, 120 g silica, 0-30% EtOAc in hexanes;
Rf=0.4 (25% EtOAc in hex)) yielded 7.6 g of benzoic
acid-cis-2-azido-trans-5-methoxycarbonyl-cyclohexyl ester as a
colorless oil. The impure solid from the previous Hex/EtOAc
filtration was mixed with 10% EtOAc in Hex and heated to reflux.
The heterogeneous solution was cooled to room temperature and the
filtrate was concentrated in vacuo to yield 5 g of a pale yellow
oil. Chromatography (Combiflash, 120 g silica, 0-30% EtOAc in
hexanes; Rf=0.4 (25% EtOAc in hex)) yielded another 1.2 g of
benzoic acid-cis-2-azido-trans-5-methoxycarbonyl-cyclohexyl ester
as a pale yellow oil (8.8 g total, 67%).
[1018] Benzoic acid-cis-2-azido-trans-5-methoxycarbonyl-cyclohexyl
ester (4.0 g, 13.2 mmol) was dissolved in MeOH (50 mL). 25% Sodium
methoxide solution (1.51 mL, 6.6 mmol) was added dropwise over 5
minutes. After 3 h, amberlite IRC-50 (8 g, weakly acidic ion
exchange resin) was added and the solution was shaken at room
temperature. After 4 h, the heterogeneous solution was filtered and
the resin was washed with MeOH (2.times.40 mL). The combined
filtrates were dried in vacuo. Chromatography (Combiflash, 120 g
silica, 5-50% EtOAc in hex) yielded 2.3 g of
trans-4-azido-trans-3-hydroxy-cyclohexane-carboxylic acid methyl
ester as a colorless oil, .sup.1H NMR 400 MHz (CDCl.sub.3) .delta.
4.05 (brs, 1H), 3.68 (s, 3H), 3.49 (m, 1H), 2.76 (m, 1H), 2.14 (m,
1H), 2.05 (m, 1H), 1.96, Brs, 1H), 1.88 m, 2H), 1.48-1.72 (m,
2H).
[1019] trans-4-Azido-trans-3-hydroxycyclohexane-carboxylic acid
methyl ester (2.75 g, 13.8 mmol) was dissolved in MeOH (70 mL) and
flushed with Ar. 10% Pd/C (0.37 g, 0.34 mmol) was added and the
flask was flushed with H.sub.2. After 18 h, the reaction was
filtered and the catalyst was washed with MeOH (2.times.10 mL). The
filtrate was concentrated and yielded a pale yellow solid which was
dissolved in DCM (60 mL) and diluted with iPr.sub.2NEt (2.65 mL,
15.2 mmol). Boc.sub.2O (3.32 g, 15.2 mmol) was added in one
portion. After 8 h, the reaction was concentrated in vacuo.
Chromatography (Combiflash, 120 g silica, 0-10% MeOH in DCM; Rf=0.4
5% MeOH in DCM) yielded 3.75 g of
trans-4-tert-butoxycarbonylamino-trans-3-hydroxy-cyclohexanecarboxylic
acid methyl ester as a white solid, .sup.1H NMR 400 MHz
(CDCl.sub.3) .delta. 4.82 (brs, 1H), 4.11H), 3.67 (s, 3H), 3.51
(brs, 1H), 2.70 (m, 1H), 2.10 (ddd, 1H, J=3.6, 6.2 and 14.1 Hz),
2.00 (m, 1H), 1.48-1.80 (m, 5H), 1.45 (s, 9 h).
[1020]
trans-4-tert-Butoxycarbonylamino-trans-3-hydroxy-cyclohexanecarboxy-
lic acid methyl ester (3.75 g, 13.7 mmol) was dissolved in DMF (20
mL) and TBSCl (2.481 g, 16.44 mmol), imidazole (2.8 g, 41.1 mmol)
and DMAP (5.02 g, 41.1 mmol) were added sequentially. After 18 h,
another portion of TBSCl (1.24 g, 8.25 mmol) was added. After 72 h,
the reaction was partitioned between Et.sub.2O (150 mL) and water
(100 mL). The layers were separated and the aqueous layer was
extracted with Et.sub.2O (2.times.100 mL). The combined organic
layers were washed with water (3.times.100 mL) and brine (50 mL),
dried over MgSO.sub.4, filtered and concentrated to yield 5.6 g of
a pale brown oil. Chromatography (Combiflash, 120 g silica, 3-15%
EtOAc in Hex; Rf 0.5 (10% EtOAc in Hex)) yielded 3.44 g of
trans-4-tert-butoxycarbonylamino-trans-3-(tert-butyl-dimethyl-silanyloxy)-
-cyclohexanecarboxylic acid methyl ester as a colorless oil which
slowly solidified.
[1021]
trans-4-tert-Butoxycarbonylamino-trans-3-(tert-butyl-dimethyl-silan-
yloxy)-cyclohexanecarboxylic acid methyl ester (3.44 g, 8.88 mmol)
was dissolved in EtOH (60 mL) and the solution was cooled to 0
degrees C. Sodium borohydride (336 mg, 8.88 mmol) was added in one
portion. The reaction was allowed to warm to room temperature
slowly overnight. After 18 h, more sodium borohydride (100 mg, 2.64
mmol) was added and the reaction was warmed to 40 degrees C. After
another 18 h, the reaction was partitioned between Et.sub.2O (200
mL) and water (100 mL). The layers were separated and the aqueous
layer was extracted with Et.sub.2O (2.times.150 mL). The combined
organic layers were washed with brine, dried over MgSO4, filtered
and concentrated to yield a colorless oil. Chromatography
(Combiflash, silica, 3%-50% EtOAc in Hex) yielded 1.2 g of
[cis-2-(tert-butyl-dimethyl-silanyloxy)trans-4-hydroxymethyl-cyclohexyl]--
carbamic acid tert-butyl ester as a colorless oil which solidifies,
.sup.1H NMR 400 MHz (CDCl.sub.3) .delta. 4.55 (d, 1H, J=9.2 Hz),
3.98 (s, 1H), 3.39 (m, 3H), 1.68-1.85 (m, 3H), 1.44-1.62 (m, 3H),
1.37 (s, 9H), 1.12 (ddd, 1H, J=1.8, 13, 13 Hz), 0.97 (m, 1H), 0.85
(s, 9H), 0.00 (s, 6H).
[1022]
[cis-2-(tert-Butyl-dimethyl-silanyloxy)-trans-4-hydroxymethyl-cyclo-
hexyl]-carbamic acid tert-butyl ester (1.33 g, 3.7 mmol) was
dissolved in dry DCM (20 mL) and cooled to 5 degrees C.
Diisopropylethylamine (0.97 mL, 5.55 mmol) and methanesulfonyl
chloride (0.344 mL, 4.44 mmol) were added sequentially. After 2 h,
the reaction was quenched with cold water (10 mL) and warmed to
room temperature. The solution was partitioned between DCM (100 mL)
and water (50 mL). The layers were separated and the aqueous layer
was extracted with DCM (2.times.100 mL). The combined organic
layers were washed with brine, dried over MgSO.sub.4, filtered and
concentrated in vacuo to yield 1.6 g (99%) of methanesulfonic acid
trans-4-tert-butoxycarbonylamino-trans-3-(tert-butyl-dimethyl-silanyloxy)-
-cyclohexylmethyl ester as a pale yellow oil. The material was used
without further purification.
[1023] Methanesulfonic acid
trans-4-tert-butoxycarbonylamino-trans-3-(tert-butyl-dimethyl-silanyloxy)-
-cyclohexylmethyl ester (1.6 g, 3.66 mmol) was dissolved in dry DMF
and sodium azide (475 mg, 7.31 mmol) was added. The reaction was
heated to 55 degrees C. After 8 h, the reaction was cooled to room
temperature and another portion of sodium azide (119 mg, 1.83 mmol)
was added. After 72 h, the reaction was partitioned between EtOAc
(100 mL) and water (50 mL). The layers were separated and the
aqueous layer was extracted with EtOAc (2.times.50 mL). The
combined organic layers were washed with water (3.times.75 mL) and
brine (15 mL), dried over MgSO.sub.4, filtered and concentrated to
yield a colorless oil. Chromatography (Combiflash, 40 g silica,
3-50% EtOAc in Hex) yielded 1.34 g (95%) of
trans-4-azidomethyl-cis-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexyl]-ca-
rbamic acid tert-butyl ester a colorless oil.
[1024]
trans-4-Azidomethyl-cis-2-(tert-butyl-dimethyl-silanyloxy)-cyclohex-
yl]-carbamic acid tert-butyl ester (1.1 g, 2.86 mmol) was dissolved
in MeOH (50 mL) and flushed with Ar. 10% Pd/C (200 mg, 0.18 mmol)
was added and the reaction was flushed with H.sub.2. After 18 h,
the reaction was flushed with Ar and the heterogeneous solution was
filtered. The catalyst was washed with MeOH (2.times.5 mL) and the
solvent was removed in vacuo to yield 900 mg of
[trans-4-aminomethyl-cis-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexyl]-c-
arbamic acid tert-butyl ester (approximately 80% pure), m/z 359.6
[M+1].sup.+. The product was used in subsequent steps without any
further purification.
[1025] A solution of diisopropylethyl amine (0.07 mL, 0.4 mmol) and
[trans-4-aminomethyl-cis-2-(tert-butyl-dimethyl-silanyloxy)-cyclohexyl]-c-
arbamic acid tert-butyl ester (290 mg, 0.81 mmol) were dissolved in
DCM (2 mL) and added to a solution of
(5-nitro-4-thiocyanato-pyrimidin-2-yl)-(2-trifluoromethoxy-benzyl)-amine
(in 3 mL of DCM). After 18 h, the reaction was partitioned between
DCM (10 mL) and water (5 mL). The layers were separated and the
water layer was extracted with DCM (2.times.5 mL). The combined
organic layers were washed with water (5 mL) and brine (5 mL),
dried over MgSO.sub.4, filtered and concentrated to yield a
colorless oil. Chromatography (Combiflash, 12 g silica, 5%-40%
EtOAc in Hex; Rf (product in 20% EtOAc in Hex) 0.3) yielded 226 mg
of
(cis-2-(tert-butyl-dimethyl-silanyloxy)-4-{[5-nitro-2-(2-trifluoromethoxy-
-benzylamino)-pyrimidin-trans-4-ylamino]-methyl}-cyclohexyl)-carbamic
acid tert-butyl ester as a pale yellow foam, m/z 671.7
[M+1].sup.+.
[1026]
(cis-2-(tert-Butyl-dimethyl-silanyloxy)-4-{[5-nitro-2-(2-trifluorom-
ethoxy-benzylamino)-pyrimidin-trans-4-ylamino]-methyl}-cyclohexyl)-carbami-
c acid tert-butyl ester (116 mg, 0.17 mmol) was dissolved in
dioxane (2 mL)/MeOH (1 mL) and 4M HCl solution (1.5 mL, 10 mmol)
was added dropwise. After 18 h, another portion of MeOH (1 mL) and
4M HCl solution (1 mL, 4 mmol) were added. After a total of 48 h,
the reaction was concentrated and the residue was triturated with
DCM. Chromatography of the residue (silica, 12 g, 9:1:2%
DCM:MeOH:NH.sub.4OH) yielded 48 mg (79%) of the title compound as a
pale yellow solid, m/z 457.5 [M+1].sup.+.
[1027] trans-4-Azido-cis-3-hydroxy-cyclohexanecarboxylic acid
methyl ester was synthesized as described previously (Krzysztof,
K., Tett. Asym., 2001, 12, 455).
Assessment of Biological Activity
PKC-theta Inhibition Assay
[1028] The ability of compounds to inhibit the kinase activity of
PKC-theta was measured using a Kinase Glo Assay. This assay detects
the enzymatic activity of protein kinase C-theta using a
firefly-luciferase reagent (Kinase-Glo--Promega #V6714) to detect
ATP levels remaining after incubation of the enzyme with ATP and
it's acceptor substrate, [Ser25]-Protein Kinase C (19-31) (Anaspec
#23020). Briefly, compounds are diluted in 100% DMSO at 100.times.
the final desired assay concentration. Compounds are subsequently
diluted 1:25 into complete assay buffer (50 mM HEPES/KOH, pH 7.5;
10 mM MgCl2; 50 mM KCl; 0.01% CHAPS; 0.1% BSA; 200 .mu.M TCEP). 10
.mu.l of the 4.times.4% DMSO stocks are transferred to 384-well
white polystyrene plates (Greiner #781075). 10 .mu.l of 4 nM
PKC-theta are added to the compounds; followed by 20 .mu.l of a
mixture containing 10 .mu.M peptide substrate and 2 .mu.M ATP.
Blank wells are defined by the addition of an equal volume of assay
buffer in place of the PKC-theta. The complete reaction is allowed
to incubate at room temperature for 90 minutes. Following this
incubation period the reaction is terminated by the addition of 40
.mu.l of the Kinase-Glo reagent. This condition is allowed to
incubate for 15 minutes after which luminescence is quantified
using an LJL Analyst.
[1029] All compounds in the synthetic examples and Tables above
were evaluated in the PKC-theta assay above and were found to have
IC.sub.50's less than 1 microM. Preferred compounds had IC.sub.50's
equal to or less than 0.01 microM.
[1030] Many of the compounds in the synthetic examples and Tables
above were also tested against Syk, Lyn, Veg-f and insulin receptor
kinase to evaluate selectivity for PKC-theta inhibition. Some
compounds were also tested against other kinases including CDK-2
and PLK. Many of the compounds demonstrated selectivity for the
inhibition of PKC-theta as compared to one or more of the other
kinases tested.
[1031] Assay conditions for testing against other kinases are
generally known in the art. Examples of suitable assays that can be
used are described below:
SYK Kinase Assay
[1032] Syk is purified as a GST-fusion protein. The kinase activity
is measured using DELFIA (Dissociation Enhanced Lanthanide
Fluoroimmunoassay) which utilizes europium chelate-labeled
anti-phosphotyrosine antibodies to detect phosphate transfer to a
random polymer, poly Glu4: Tyr1 (PGTYR).
[1033] The kinase assay is performed in kinase assay buffer (50 mM
HEPES, pH 7.0, 25 mM MgCl2, 5 mM MnCl2, 50 mM KCl, 100 .mu.M
Na3VO4, 0.2% BSA, 0.01% CHAPS, 200 .mu.M TCEP). Test compounds
initially dissolved in DMSO at 5 mg/mL, are prediluted for dose
response (starting conc. 10 .mu.M (or 5 .mu.g/mL), 1 to 3 serial
dilutions, 10 doses) with the assay buffer in 96-well polypropylene
microtiter plates. A 40 .mu.L volume of diluted enzyme (0.5 nM
final conc.) in kinase buffer and a 20 .mu.L aliquot of diluted
compound are sequentially added to neutravidin coated 96-well white
plate (PIERCE). The kinase reaction is started with a 40 .mu.L
volume of a mixture of substrates containing 0.75 .mu.M ATP plus
4.5 ng/.mu.L PGTYR-biotin (CIS Biointernational) in kinase buffer.
Background wells are incubated with kinase plus buffer, and the
reference inhibitor wells are incubated with 20 .mu.L of 25 .mu.M
ADP instead of the compound. The assay plates are incubated for 30
min at room temperature. Following incubation, the assay plates are
washed three times with 250 .mu.L wash buffer (50 mM Tris-HCL, pH
7.4, 150 mM NaCl, 0.05% Tween 20, 0.2% BSA). A 100 .mu.L aliquot of
europium-labeled anti-phosphotyrosine (Eu3+-PT66, Wallac CR04-100)
diluted in 50 mM Tris-HCl, pH 7.8, 150 mM NaCl, 10 .mu.M DTPA,
0.05% Tween 40, 0.2% BSA, 0.05% BGG (1 nM final conc.) is added to
each well and incubated for 30 min at room temperature. Upon
completion of the incubation, the plate is washed four times with
250 .mu.L of wash buffer and 100 .mu.L of DELFIA Enhancement
Solution (Wallac) is added to each well. After 15 min or longer,
time-resolved fluorescence is measured on the LJL's Analyst
(excitation at 360 nm, emission at 620 nm, EU 400 Dicbroic Mirror)
after a delay time of 250 .mu.s.
LYN Kinase Assay
[1034] Lyn(Kd) is purified as a GST-fusion protein. The kinase
activity is measured using DELFIA (Dissociation Enhanced Lanthanide
Fluoroimmunoassay) which utilizes europium chelate-labeled
anti-phosphotyrosine antibodies to detect phosphate transfer to a
random polymer, poly Glu4: Tyr1 (PGTYR).
[1035] The kinase assay is performed in kinase assay buffer (50 mM
HEPES, pH 7.0, 25 mM MgCl2, 5 mM MnCl2, 50 mM KCl, 100 .mu.M
Na3VO4, 0.2% BSA, 0.01% CHAPS, 200 .mu.M TCEP). Test compounds
initially dissolved in DMSO at 5 mg/mL, are pre-diluted for dose
response (starting conc. 10 .mu.M (or 5 .mu.g/mL), 1 to 3 serial
dilutions, 10 doses) with the assay buffer in 96-well polypropylene
microtiter plates. A 40 .mu.L volume of diluted enzyme (0.7 nM
final conc.) in kinase buffer and a 20 .mu.L aliquot of diluted
compound are sequentially added to neutravidin coated 96-well white
plate (PIERCE). The kinase reaction is started with a 40 .mu.L
volume of a mixture of substrates containing 1.25 .mu.M ATP plus
4.5 ng/.mu.L PGTYR-biotin (CIS Biointernational) in kinase buffer.
Background wells are incubated with kinase plus buffer, and the
reference inhibitor wells are incubated with 20 .mu.L of 25 .mu.M
ADP instead of the compound. The assay plates are incubated for 30
min at room temperature. Following incubation, the assay plates are
washed three times with 250 .mu.L wash buffer (50 mM Tris-HCL, pH
7.4, 150 mM NaCl, 0.05% Tween 20, 0.2% BSA). A 100 .mu.L aliquot of
europium-labeled anti-phosphotyrosine (Eu3+-PT66, Wallac CR04-100)
diluted in 50 mM Tris-HCl, pH 7.8, 150 mM NaCl, 10 .mu.M DTPA,
0.05% Tween 40, 0.2% BSA, 0.05% BGG (1 nM final conc.) is added to
each well and incubated for 30 min at room temperature. Upon
completion of the incubation, the plate is washed four times with
250 .mu.L of wash buffer and 100 .mu.L of DELFIA Enhancement
Solution (Wallac) is added to each well. After 15 min or longer,
time-resolved fluorescence is measured on the LJL's Analyst
(excitation at 360 nm, emission at 620 nm, EU 400 Dichroic Mirror)
after a delay time of 250 .mu.s.
Methods of Therapeutic Use
[1036] The compounds of the invention are effective inhibitors of
PKC-theta activity, and therefore are useful to inhibit PKC-theta
activity in a patient and treat a variety of diseases and disorders
that are mediated or sustained through the activity of
PKC-theta.
[1037] Without wishing to be bound by theory, the compounds of this
invention would be expected to inhibit T cell activation via
effective inhibition of PKC-theta, and are therefore useful to
treat diseases and disorders associated with T cell activation. For
example, the inhibition of T cell activation is therapeutically
useful for selectively suppressing the immune function. Thus, the
inhibition of PKC-theta with the compounds of this invention is an
attractive means for treating a variety of immunological disorders,
including inflammatory diseases, autoimmune diseases, organ and
bone marrow transplant rejection and other disorders associated
with T cell mediated immune response. In particular, the compounds
of the invention may be used to treat acute or chronic
inflammation, allergies, contact dermatitis, psoriasis, rheumatoid
arthritis, multiple sclerosis, type I diabetes, inflammatory bowel
disease, Guillain-Barre syndrome, Crohn's disease, ulcerative
colitis, graft versus host disease (and other forms of organ or
bone marrow transplant rejection) and lupus erythematosus. Other
disorders associated with T cell-mediated immune responses will be
evident to those of ordinary skill in the art and can also be
treated with the compounds and compositions of this invention.
[1038] In addition, PKC theta activation has been shown to be
associated with insulin resistance in skeletal muscle. Therefore,
the inhibition of PKC-theta with the compounds of this invention is
also an attractive means for treating type II diabetes.
[1039] For therapeutic use, the compounds of the invention may be
administered via a pharmaceutical composition in any conventional
pharmaceutical dosage form in any conventional manner. Conventional
dosage forms typically include a pharmaceutically acceptable
carrier suitable to the particular dosage form selected. Routes of
administration include, but are not limited to, intravenously,
intramuscularly, subcutaneously, intrasynovially, by infusion,
sublingually, transdermally, orally, topically or by inhalation.
The preferred modes of administration are oral and intravenous.
[1040] The compounds of this invention may be administered alone or
in combination with adjuvants that enhance stability of the
inhibitors, facilitate administration of pharmaceutical
compositions containing them in certain embodiments, provide
increased dissolution or dispersion, increase inhibitory activity,
provide adjunct therapy, and the like, including other active
ingredients. In one embodiment, for example, multiple compounds of
the present invention can be administered. Advantageously, such
combination therapies utilize lower dosages of the conventional
therapeutics, thus avoiding possible toxicity and adverse side
effects incurred when those agents are used as monotherapies.
Compounds of the invention may be physically combined with the
conventional therapeutics or other adjuvants into a single
pharmaceutical composition. Advantageously, the compounds may then
be administered together in a single dosage form. In some
embodiments, the pharmaceutical compositions comprising such
combinations of compounds contain at least about 5%, but more
preferably at least about 20%, of a compound of formula (I) (w/w)
or a combination thereof. The optimum percentage (w/w) of a
compound of the invention may vary and is within the purview of
those skilled in the art. Alternatively, the compounds of the
present invention and the conventional therapeutics or other
adjuvants may be administered separately (either serially or in
parallel). Separate dosing allows for greater flexibility in the
dosing regime.
[1041] As mentioned above, dosage forms of the compounds of this
invention may include pharmaceutically acceptable carriers and
adjuvants known to those of ordinary skill in the art and suitable
to the dosage form. These carriers and adjuvants include, for
example, ion exchangers, alumina, aluminum stearate, lecithin,
serum proteins, buffer substances, water, salts or electrolytes and
cellulose-based substances. Preferred dosage forms include tablet,
capsule, caplet, liquid, solution, suspension, emulsion, lozenges,
syrup, reconstitutable powder, granule, suppository and transdermal
patch. Methods for preparing such dosage forms are known (see, for
example, H. C. Ansel and N. G. Popovish, Pharmaceutical Dosage
Forms and Drug Delivery Systems, 5th ed., Lea and Febiger (1990)).
Dosage levels and requirements for the compounds of the present
invention may be selected by those of ordinary skill in the art
from available methods and techniques suitable for a particular
patient. In some embodiments, dosage levels range from about 1-1000
mg/dose for a 70 kg patient. Although one dose per day may be
sufficient, up to 5 doses per day may be given. For oral doses, up
to 2000 mg/day may be required. As the skilled artisan will
appreciate, lower or higher doses may be required depending on
particular factors. For instance, specific dosage and treatment
regimens will depend on factors such as the patient's general
health profile, the severity and course of the patient's disorder
or disposition thereto, and the judgment of the treating
physician.
* * * * *