U.S. patent application number 10/822411 was filed with the patent office on 2004-10-07 for compounds for the treatment of ischemia.
Invention is credited to DeNinno, Michael P., Masamune, Hiroko, Scott, Robert W..
Application Number | 20040198693 10/822411 |
Document ID | / |
Family ID | 33100715 |
Filed Date | 2004-10-07 |
United States Patent
Application |
20040198693 |
Kind Code |
A1 |
DeNinno, Michael P. ; et
al. |
October 7, 2004 |
Compounds for the treatment of ischemia
Abstract
A.sub.3 agonists, methods of using such A.sub.3 agonists and
pharmaceutical compositions containing such A.sub.3 agonists. The
A.sub.3 agonists are useful for the reduction of tissue damage
resulting from tissue ischemia or hypoxia.
Inventors: |
DeNinno, Michael P.; (Gales
Ferry, CT) ; Masamune, Hiroko; (Noank, CT) ;
Scott, Robert W.; (Mystic, CT) |
Correspondence
Address: |
SCULLY SCOTT MURPHY & PRESSER, PC
400 GARDEN CITY PLAZA
GARDEN CITY
NY
11530
|
Family ID: |
33100715 |
Appl. No.: |
10/822411 |
Filed: |
April 12, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10822411 |
Apr 12, 2004 |
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09640530 |
Aug 17, 2000 |
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60156828 |
Sep 30, 1999 |
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Current U.S.
Class: |
514/46 ;
514/263.23; 514/263.4; 514/303; 536/27.3; 544/277; 546/119 |
Current CPC
Class: |
C07D 473/30 20130101;
C07D 471/04 20130101; C07H 19/16 20130101; C07D 473/00
20130101 |
Class at
Publication: |
514/046 ;
514/303; 544/277; 546/119; 514/263.23; 536/027.3; 514/263.4 |
International
Class: |
C07H 019/16; C07D
487/14; C07D 473/14; A61K 031/52 |
Claims
1. A compound of the formula I 18a prodrug thereof or a
pharmaceutically acceptable salt of said compound or of said
prodrug, wherein X is oxy, methylene or thio; Y is CH or N; Z is H,
(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkyloxy, trifluoromethyl
or halo; R.sup.1 is hydroxymethyl, (C.sub.1-C.sub.3)alkoxymethyl,
(C.sub.3-C.sub.5)cycloalkox- ymethyl, carboxy,
(C.sub.1-C.sub.3)alkoxycarbonyl, (C.sub.3-C.sub.5)cycloa-
lkoxycarbonyl, 1,1-aminoiminomethyl, 1,1-(mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino)iminomethyl, 1,1-(mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylamino)iminomethyl, carbamoyl,
mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl or
N-(C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl;
R.sup.2 is H, (C.sub.1-C.sub.3)alkyl or
(C.sub.3-C.sub.5)cycloalkyl; R.sup.3 is halo, trifluoromethyl,
cyano, (C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)lkyloxy, ethenyl or
ethynyl; D is oxy, thio, NH, (C.sub.1-C.sub.6)alkyloxy,
(C.sub.1-C.sub.6)alkylthio or (C.sub.1-C.sub.6)alkylamino; G is a
partially saturated, fully saturated or fully unsaturated five to
eight membered ring optionally having one to three heteroatoms
selected independently from oxygen, sulfur and nitrogen, or, a
bicyclic ring consisting of two fused partially saturated, fully
saturated or fully unsaturated three to six membered rings, taken
independently, optionally having one to four heteroatoms selected
independently from nitrogen, sulfur and oxygen; wherein said G is
optionally mono- , di- or tri-substituted independently with halo,
(C.sub.1-C.sub.3)alkyl, trifluoromethyl, trifluoromethoxy, nitro,
cyano, (C.sub.3-C.sub.5)cycloalkyl, hydroxy or
(C.sub.1-C.sub.3)alkoxy or G is cyano,
(C.sub.1-C.sub.4)alkoxycarbonyl, (C.sub.3-C.sub.5)
cycloalkoxycarbonyl, C(O)NR.sup.4R.sup.5, C(S)NR.sup.4R.sup.5,
C(NH)NR.sup.4R.sup.5, C(N(C.sub.1-C.sub.3)alkyl)NR.sup.4R.sup.5 or
C(N(C.sub.3-C.sub.10)cycloalkyl)NR.sup.4R.sup.5; R.sup.4 is a bond,
H, (C.sub.1-C.sub.10)alkyl, hydroxy, (C.sub.1-C.sub.10)alkoxy,
(C.sub.3-C.sub.10)cycloalkoxy or a partially saturated, fully
saturated or fully unsaturated five to eight membered ring,
optionally linked through (C.sub.1-C.sub.3)alkyl, optionally having
one to three heteroatoms selected independently from oxygen, sulfur
and nitrogen, or, a bicyclic ring or a bicyclic ring with optional
(C.sub.1-C.sub.3)bridge optionally linked through
(C.sub.1-C.sub.3)alkyl, said bicyclic ring or bridged bicyclic ring
optionally having one to four heteroatoms selected independently
from nitrogen, sulfur and oxygen wherein said
(C.sub.1-C.sub.10)alkyl, (C.sub.1-C.sub.10)alkoxy,
(C.sub.3-C.sub.10)cycloalkoxy or R.sup.4 ring(s) is optionally
mono-, di- or tri-substituted independently with halo,
(C.sub.1-C.sub.3)alkyl, trifluoromethyl, nitro, cyano,
(C.sub.3-C.sub.5) cycloalkyl, hydroxy or (C.sub.1-C.sub.3) alkoxy;
R.sup.5 is a bond, H, (C.sub.1-C.sub.10)alkyl or
(C.sub.1-C.sub.10)cycloalkyl; or R.sup.4 and R.sup.5 taken together
with the nitrogen to which they are attached form a fully saturated
or partially unsaturated four to nine membered ring, said ring
optionally bridged, optionally having one to three heteroatoms
selected independently from oxygen, sulfur and nitrogen, said ring
optionally mono- or di-substituted independently with oxo, hydroxy,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.8) alkyl, amino, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
N-(C.sub.1-C.sub.4)alkyl-
-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cy- cloalkylamin, N-(
C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylamino- ,
formylamino, (C.sub.1-C.sub.4)alkylcarbonylamino,
(C.sub.3-C.sub.5)cycloalkylcarbonylamino,
(C.sub.1-C.sub.4)alkoxycarbonyl- amino,
N-(C.sub.1-C.sub.4)alkoxycarbonyl-N-(C.sub.1-C.sub.4)alkylamino,
(C.sub.1-C.sub.4)sulfamoyl, (C.sub.1-C.sub.4)alkylsulfonylamino,
(C.sub.3-C.sub.5)cycloalkylsulfonylamino or a partially saturated,
fully saturated or fully unsaturated five to eight membered ring,
optionally linked through (C.sub.1-C.sub.3)alkyl, optionally having
one to three heteroatoms selected independently from oxygen, sulfur
and nitrogen, or, a bicyclic ring consisting of two fused partially
saturated, fully saturated or fully unsaturated three to six
membered rings, taken independently, optionally linked through
(C.sub.1-C.sub.3)alkyl, optionally having one to four heteroatoms
selected independently from nitrogen, sulfur and oxygen, and
optionally mono- or di-substituted with halo, trifluoromethyl,
trifluoromethoxy, (C.sub.1-C.sub.3)alkyl or
(C.sub.1-C.sub.3)alkoxy.
2. A compound as recited in claim 1 wherein X is oxy; Y is N; Z is
H; R.sup.1 is (C.sub.1-C.sub.6)alkylcarbamoyl; R.sup.2 is H;
R.sup.3 is halo, trifluoromethyl, cyano, (C.sub.1-C.sub.3)alkyl,
(C.sub.1-C.sub.3)alkyloxy, ethenyl or ethynyl; D is oxy, thio,
(C.sub.1-C.sub.6)alkyloxy or (C.sub.1-C.sub.6)alkylthio; G is
phenyl, pyridyl, pyrimidinyl, pyrazinyl, thiazolyl, oxazolyl,
isoxazolyl, pyridazinyl, tetrazolyl, isothiazolyl, thiophenyl,
furanyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl, pyrazolyl,
pyrrolyl, indolyl, naphthalenyl, quinolinyl, isoquinolinyl,
benzo[b]furanyl, benzo[b]thiophenyl, benzothiazolyl,
tetrahydrofuranyl, pyrrolidinyl, piperidinyl, tetrahydropyranyl,
morpholinyl wherein said G is optionally mono-, di- or
tri-substituted independently with halo, (d-C.sub.3)alkyl or
(d-C.sub.3)alkoxy, or a pharmaceutically acceptable salt
thereof.
3. A compound as recited in claim 2 wherein R.sup.1 is
methylcarbamoyl; R.sup.3 is halo; D is (C.sub.1-C.sub.6)alkoxy; G
is phenyl, pyridyl, thiazolyl, oxazolyl, isoxazolyl, isothiazolyl,
furanyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl, pyrazolyl, pyrrolyl
wherein said G is optionally mono-, di- or tri-substituted
independently with halo, (C.sub.1-C.sub.3)alkyl, trifluoromethoxy
or (C.sub.1-C.sub.3)alkoxy, or a pharmaceutically acceptable salt
thereof.
4. A compound as recited in claim 3 wherein D is
(C.sub.1-C.sub.2)alkoxy; G is phenyl, thiazolyl, oxazolyl,
isoxazolyl, tetrahydrofuranyl, pyrrolidinyl, piperidinyl or
morpholinyl wherein said G is optionally mono-, di- or
tri-substituted independently with halo, (C.sub.1-C.sub.3)alkyl or
(C.sub.1-C.sub.3)alkoxy, or a pharmaceutically acceptable salt
thereof.
5. A compound as recited in claim 3 wherein R.sup.3 is chloro; D is
methyleneoxy; and G is phenyl, or a pharmaceutically acceptable
salt thereof.
6. A compound as recited in claim 3 wherein R.sup.3 is chloro; D is
methyleneoxy; and G is 3-furanyl, or a pharmaceutically acceptable
salt thereof.
7. A compound as recited in claim 3 wherein R.sup.3 is chloro; D is
methyleneoxy; and G is 2-furanyl, or a pharmaceutically acceptable
salt thereof.
8. A compound as recited in claim 3 wherein R.sup.3 is chloro; D is
methyleneoxy; and G is 2-thiazolyl, or a pharmaceutically
acceptable salt thereof.
9. A compound as recited in claim 3 wherein R.sup.3 is chloro; D is
methyleneoxy; and G is 5-(3-methylisoxazolyl), or a
pharmaceutically acceptable salt thereof.
10. A compound as recited in claim 1 wherein X is oxy; Y is N; Z is
H; R.sup.1 is (C.sub.1-C.sub.6)alkylcarbamoyl; R.sup.2 is H;
R.sup.3 is halo, trifluoromethyl, cyano, (C.sub.1-C.sub.3)alkyl,
(C.sub.1-C.sub.3)alkyloxy, ethenyl or ethynyl; D is
(C.sub.1-C.sub.6)alkyloxy or (C.sub.1-C.sub.6)alkylthio; G is
C(O)NR.sup.4R.sup.5or C(S)NR.sup.4 R.sup.5 wherein R.sup.4 and
R.sup.5 taken together with the nitrogen to which they are attached
form a fully saturated four to nine membered ring, optionally
having one to three additional heteroatoms selected independently
from oxygen, sulfur and nitrogen, said ring optionally mono- or
di-substituted independently with oxo, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.8)alkyl, amino, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
N-(C.sub.1-C.sub.4)alkyl-
-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl, mono-N- or
di-N,N-(d-C.sub.4)alkylamino, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalk- ylamino or
N-(C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylamino,
formylamino, (C.sub.1-C.sub.4)alkylformylamino,
(C.sub.3-C.sub.5)cycloalk- ylformylamino, sulfamoyl,
(Crd)alkylsulfonylamino, (C.sub.3-C.sub.5)cycloa- lkylsulfonylamino
or a partially saturated, fully saturated or fully unsaturated five
to eight membered ring, optionally linked through
(C.sub.1-C.sub.3)alky'l, optionally having one to three heteroatoms
selected independently from oxygen, sulfur and nitrogen, or, a
bicyclic ring consisting of two fused partially saturated, fully
saturated or fully unsaturated three to six membered rings, taken
independently, optionally linked through (C.sub.1-C.sub.3)alkyl,
optionally having one to four heteroatoms selected independently
from nitrogen, sulfur and oxygen, or a pharmaceutically acceptable
salt thereof.
11. A compound as recited in claim 10 wherein R.sup.1 is
methylcarbamoyl; R.sup.3 is halo; D is (C.sub.1-C.sub.2)alkoxy; G
is C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4 R.sup.5; wherein R.sup.4 and
R.sup.5 taken together with the nitrogen to which they are attached
form piperidinyl, piperazinyl, morpholinyl, azetidinyl or
pyrrolidinyl said ring optionally mono- or di-substituted
independently with oxo, hydroxy, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.8)alkyl, amino, carbamoyl, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
N-(C.sub.1-C.sub.4)alkyl-
-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cy- cloalkylamino or
N-(C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylami- no,
formylamino, (C.sub.1-C.sub.4)alkylformylamino,
(C.sub.3-C.sub.5)cycloalkylformylamino, sulfamoyl,
(C.sub.1-C.sub.4)alkylsulfonylamino,
(C.sub.3-C.sub.5)cycloalkylsulfonyla- mino or a partially
saturated, fully saturated or fully unsaturated four to eight
membered ring, optionally linked through (C.sub.1-C.sub.3)alkyl,
optionally having one to two heteroatoms selected independently
from oxygen, sulfur and nitrogen, or a pharmaceutically acceptable
salt thereof.
12. A compound as recited in claim 11 wherein G is
C(O)NR.sup.4R.sup.5; wherein R.sup.4 and R.sup.5 taken together
with the nitrogen to which they are attached form piperidinyl,
piperazinyl, morpholinyl, azetidinyl, pyrrolidinyl said ring
optionally mono- or di-substituted independently with hydroxy,
oxo,(C.sub.1-C.sub.6)alkoxy, (C.sub.1C.sub.8)alkyl, amino,
carbamoyl, mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl,
mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylamino, or a partially
saturated, fully saturated or fully unsaturated four to eight
membered ring, optionally linked through (C.sub.1-C.sub.3)alkyl,
optionally having one to two heteroatoms selected independently
from oxygen, sulfur and nitrogen, or a pharmaceutically acceptable
salt thereof.
13. A compound as recited in claim 12 wherein R.sup.3 is chloro; D
is methyleneoxy; G is C(O)NR.sup.4R.sup.5; wherein R.sup.4 and
R.sup.5 taken together with the nitrogen to which they are attached
form piperazinyl substituted in the four position with methyl, or a
pharmaceutically acceptable salt thereof.
14. A compound as recited in claim 12 wherein R.sup.3 is chloro; D
is methyleneoxy; G is C(O)NR.sup.4R.sup.5; wherein R.sup.4 and
R.sup.5 taken together with the nitrogen to which they are attached
form piperazinyl, or a pharmaceutically acceptable salt
thereof.
15. A compound as recited in claim 12 wherein R.sup.3 is chloro; D
is methyleneoxy; G is C(O)NR.sup.4R.sup.5; wherein R.sup.4 and
Retaken together with the nitrogen to which they are attached form
piperidinyl substituted in the four position with
N,N-dimethylamino, or a pharmaceutically acceptable salt
thereof.
16. A compound as recited in claim 12 wherein R.sup.3 is chloro; D
is methyleneoxy; G is C(O)NR.sup.4R.sup.5; wherein R.sup.4 and
R.sup.5 taken together with the nitrogen to which they are attached
form piperidinyl substituted in the four position with
piperidin-1-yl, or a pharmaceutically acceptable salt thereof.
17. A compound as recited in claim 12 wherein R.sup.3 is chloro; D
is methyleneoxy; G is C(O)NR.sup.4R.sup.5; wherein R.sup.4 and
R.sup.5 taken together with the nitrogen to which they are attached
form piperidinyl substituted in the four position with methylamino,
or a pharmaceutically acceptable salt thereof.
18. A compound as recited in claim 1 wherein X is oxy; Y is N;
ZisH; R.sup.1 is (C.sub.1-C.sub.6)alkylcarbamoyl; R.sup.2 is H;
R.sup.3 is halo, trifluoromethyl, cyano, (C.sub.1-C.sub.3)alkyl,
(C.sub.1-C.sub.3)alkyloxy, ethenyl or ethynyl; D is
(C.sub.1-C.sub.6)alkyloxy or (C.sub.1-C.sub.6)alkylthio; G is
C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4R.sup.5 R.sup.4 is H,
(C.sub.1-C.sub.10)alkyl, hydroxy, (C.sub.1-C.sub.10)alkoxy,
(C.sub.3-C.sub.10)cycloalkoxy or a partially saturated, fully
saturated or fully unsaturated five to eight membered ring,
optionally linked through (C.sub.1-C.sub.3)alkyl, optionally having
one to three heteroatoms selected independently from oxygen, sulfur
and nitrogen, or, a bicyclic ring consisting of two fused partially
saturated, fully saturated or fully unsaturated three to six
membered rings, taken independently, optionally linked through
(C.sub.1-C.sub.3)alkyl, optionally having one to four heteroatoms
selected independently from nitrogen, sulfur and oxygen; R.sup.5 is
H, (C.sub.1C.sub.10)alkyl or (C.sub.1-C.sub.10)cycloalkyl, or a
pharmaceutically acceptable salt thereof.
19. A compound as recited in claim 18 wherein R.sup.1 is
methylcarbamoyl; R.sup.3 is halo; D is (C.sub.1-C.sub.2)alkoxy; G
is C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4R.sup.5; R.sup.4 is H,
(C.sub.1-C.sub.10)alkyl hydroxy, (C.sub.1-C.sub.10)alkoxy,
(C.sub.3-C.sub.10)cycloalkoxy or a partially saturated, fully
saturated or fully unsaturated five to eight membered ring,
optionally linked through (C.sub.1-C.sub.3)alkyl, optionally having
one to three heteroatoms selected independently from oxygen, sulfur
and nitrogen; and R.sup.5 is H, (C.sub.1-C.sub.10)alkyl or
(C.sub.1-C.sub.10)cycloalkyl, or a pharmaceutically acceptable salt
thereof.
20. A compound as recited in claim 19 wherein G is
C(O)NR.sup.4R.sup.5R.su- p.4 is H, (C.sub.1-C.sub.10)alkyl,
(C.sub.3-C.sub.6)cycloalkyl, hydroxy, (C.sub.1-C.sub.10)alkoxy or
(C.sub.3-C.sub.10)cycloalkoxy; and R.sup.5 is H,
(C.sub.1-C.sub.10)alkyl or (C.sub.3-C.sub.10)cycloalkyl, or a
pharmaceutically acceptable salt thereof.
21. A compound as recited in claim 20 wherein R.sup.3 is chloro; D
is methyleneoxy; G is C(O)NR.sup.4R.sup.5; R.sup.4 is H; and
R.sup.5 is H, or a pharmaceutically acceptable salt thereof.
22. A compound as recited in claim 1 wherein D is oxy, thio,
(C.sub.1-C.sub.6)alkyloxy or (C.sub.1-C.sub.6)alkylthio; G is
phenyl, pyridyl, pyrimidinyl, pyrazinyl, thiazolyl, oxazolyl,
isoxazolyl, pyridazinyl, tetrazolyl, isothiazolyl, thiophenyl,
furanyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl, pyrazolyl,
pyrrolyl, indolyl, naphthalenyl, quinolinyl, isoquinolinyl,
benzo[b]furanyl, benzo[b]thiophenyl, benzothiazolyl,
tetrahydrofuranyl, pyrrolidinyl, piperidinyl, tetrahydropyranyl,
morpholinyl wherein said G is optionally mono-, di- or
tri-substituted independently with halo, (C.sub.1-C.sub.3)alkyl or
(C.sub.1-C.sub.3)alkoxy, or a pharmaceutically acceptable salt
thereof.
23. A compound as recited in claim 22 wherein D is
(C.sub.1-C.sub.6)alkoxy- ; G is phenyl, pyridyl, thiazolyl,
oxazolyl, isoxazolyl, isothiazolyl, furanyl, 1,2,4-oxadiazolyl,
1,2,4-thiadiazolyl, pyrazolyl, pyrrolyl wherein said G is
optionally mono-, di- or tri-substituted independently with halo,
(C.sub.1-C.sub.3)alkyl or (C.sub.1-C.sub.3)alkoxy, or a
pharmaceutically acceptable salt thereof.
24. A compound as recited in claim 1 wherein D is
(C.sub.1-C.sub.6)alkylox- y or (C.sub.1-C.sub.6)alkylthio; G is
C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4R.sup.5 wherein R.sup.4 and
R.sup.5 taken together with the nitrogen to which they are attached
form a fully saturated four to nine membered ring, optionally
having one to three additional heteroatoms selected independently
from oxygen, sulfur and nitrogen, said ring optionally mono- or
di-substituted independently with oxo, hydroxy,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.8)alkyl, amino, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
N-(C.sub.1-C.sub.4)alkyl-
-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cy- cloalkylamino or
N-(C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylami- no,
formylamino, (C.sub.1-C.sub.4) alkylformylamino,
(C.sub.3-C.sub.5)cycloalkylfornylamino, sulfamoyl,
(C.sub.1-C.sub.4)alkylsulfonylamino,
(C.sub.3-C.sub.5)cycloalkylsulfonyla- mino or a partially
saturated, fully saturated or fully unsaturated five to eight
membered ring, optionally linked through (C.sub.1-C.sub.3)alkyl,
optionally having one to three heteroatoms selected independently
from oxygen, sulfur and nitrogen, or, a bicyclic ring consisting of
two fused partially saturated, fully saturated or fully unsaturated
three to six membered rings, taken independently, optionally linked
through (C.sub.1-C.sub.3)alkyl, optionally having one to four
heteroatoms selected independently from nitrogen, sulfur and
oxygen, or a pharmaceutically acceptable salt thereof.
25. A compound as recited in claim 24 wherein D is
(C.sub.1-C.sub.2)alkoxy- ; G is C(O)NR.sup.4R.sup.5 or
C(S)NR.sup.4R.sup.5; wherein R.sup.4 and R.sup.5 taken together
with the nitrogen to which they are attached form piperidinyl,
piperazinyl, morpholinyl, azetidinyl or pyrrolidinyl said ring
optionally mono- or di-substituted independently with oxo,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.8)alkyl, amino, carbamoyl,
mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
N-(C.sub.1-C.sub.4)alkyl-
-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cy- cloalkylamino or
N-(C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylami- no,
formylamino, (C.sub.1-C.sub.4)alkylformylamino,
(C.sub.3-C.sub.5)cycloalkylformylamino, sulfamoyl,
(C.sub.1-C.sub.4)alkylsulfonylamino,
(C.sub.3-C.sub.5)cycloalkylsulfonyla- mino or a partially
saturated, fully saturated or fully unsaturated four to eight
membered ring, optionally linked through (C.sub.1-C.sub.3)alkyl,
optionally having one to two heteroatoms selected independently
from oxygen, sulfur and nitrogen, or a pharmaceutically acceptable
salt thereof.
26. A compound as recited in claim 25 wherein G is
C(O)NR.sup.4R.sup.5; wherein R.sup.4 and R.sup.5 taken together
with the nitrogen to which they are attached form piperidinyl,
piperazinyl, morpholinyl, azetidinyl, pyrrolidinyl said ring
optionally mono- or di-substituted independently with oxo, hydroxy,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.8)alkyl, amino, carbamoyl,
mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino, or a partially saturated, fully
saturated or fully unsaturated four to eight membered ring,
optionally linked through (C.sub.1-C.sub.3)alkyl, optionally having
one to two heteroatoms selected independently from oxygen, sulfur
and nitrogen, or a pharmaceutically acceptable salt thereof.
27. A compound as recited in claim 1 wherein D is
(C.sub.1-C.sub.6)alkylox- y or (C.sub.1-C.sub.6)alkylthio; G is
C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4R.sup.5; R.sup.4 is H,
(C.sub.1-C.sub.10)alkyl, hydroxy, (C.sub.1-C.sub.10)alkoxy,
(C.sub.3-C.sub.10)cycloalkoxy or a partially saturated, fully
saturated or fully unsaturated five to eight membered ring,
optionally linked through (C.sub.1-C.sub.3)alkyl, optionally having
one to three heteroatoms selected independently from oxygen, sulfur
and nitrogen, or, a bicyclic ring consisting of two fused partially
saturated, fully saturated or fully unsaturated three to six
membered rings, taken independently, optionally linked through
(C.sub.1-C.sub.3)alkyl, optionally having one to four heteroatoms
selected independently from nitrogen, sulfur and oxygen; R.sup.5 is
H, (C.sub.1-C.sub.10)alkyl or (C.sub.1-C.sub.10)cycloalkyl, or a
pharmaceutically acceptable salt thereof.
28. A compound as recited in claim 27 wherein D is
(C.sub.1-C.sub.2)alkoxy- ; R.sup.4 is H, (C.sub.1-C.sub.10)alkyl,
hydroxy, (C.sub.1C.sub.10)alkoxy, (C.sub.3-C.sub.10)cycloalkoxy or
a partially saturated, fully saturated or fully unsaturated five to
eight membered ring, optionally linked through
(C.sub.1-C.sub.3)alkyl, optionally having one to three heteroatoms
selected independently from oxygen, sulfur and nitrogen; and
R.sup.5 is H, (C.sub.1-C.sub.10)alkyl or
(C.sub.1-C.sub.10)cycloalkyl, or a pharmaceutically acceptable salt
thereof.
29. A compound as recited in claim 28 wherein G is
C(O)NR.sup.4R.sup.5; R.sup.4 is H, (C.sub.1-C.sub.10)alkyl,
(C.sub.3-C.sub.6)cycloalkyl, hydroxy, (C.sub.1-C.sub.10)alkoxy or
(C.sub.3-C.sub.10)cycloalkoxy; and R.sup.5 is H,
(C.sub.1-C.sub.10)alkyl or (C.sub.3-C.sub.10)cycloalkyl, or a
pharmaceutically acceptable salt thereof.
30. A compound having the Formula C 19wherein R.sup.20 and R.sup.21
are each independently (C.sub.1-C.sub.4)alkyl, H, phenyl,
phenyl(C.sub.1-C.sub.4)alkyl or are joined together to form a
piperidinyl, pyrrolidinyl or morpholinyl ring; R.sup.22 and
R.sup.23 are each independently (C.sub.1-C.sub.4)alkyl or are
joined together to form a 5-6 membered carbocyclic ring; and
R.sup.24 is (C.sub.1-C.sub.4)alkyl, phenyl or
phenyl(C.sub.1-C.sub.4)alkyl, said phenyl or
phenyl(C.sub.1-C.sub.4)alkyl optionally mono-, di, or
tri-substituted independently on the phenyl moiety with nitro, halo
or trifluoromethyl.
31. A compound having the Formula CI 20wherein R.sup.20 and
R.sup.21 are each independently (C.sub.1-C.sub.4)alkyl, H, phenyl,
phenyl(C.sub.1-C.sub.4)alkyl or are joined together to form a
piperidinyl, pyrrolidinyl or morpholinyl ring; and R.sup.24 is
(C.sub.1-C.sub.4)alkyl, phenyl or phenyl (C.sub.1-C.sub.4)alkyl,
said phenyl or phenyl(d-d)alkyl optionally mono-, di, or
tri-substituted independently on the phenyl moiety with nitro, halo
or trifluoromethyl.
32. A compound having the Formula CII 21wherein R.sup.20 and
R.sup.21 are each independently (d-C.sub.4)alkyl, H, phenyl,
phenyl(d-C.sub.4)alkyl or are joined together to form a
piperidinyl, pyrrolidinyl or morpholinyl ring; R.sup.24 is
(C.sub.1-C.sub.4)alkyl, phenyl or phenyl (C.sub.1-C.sub.4)alkyl,
said phenyl or pheny(C.sub.1-C.sub.4)alkyl optionally mono-, di, or
tri-substituted independently on the phenyl moiety with nitro, halo
or trifluoromethyl; and R.sup.25 and R.sup.26 are independently
(C.sub.1-C.sub.4)alkyl or phenyl.
33. A compound having the Formula CIII 22wherein R.sup.3 is halo,
trifluoromethyl, cyano, (C.sub.1-C.sub.3)alkyl,
(C.sub.1-C.sub.3)alkyloxy- , ethenyl or ethynyl.
34. A compound as recited in claim 33 wherein R.sup.3 is
trifluoromethyl.
35. A compound as recited in claim 33 wherein R.sup.3 is
fluoro.
36. A compound as recited in claim 33 wherein R.sup.3 is
chloro.
37. A compound having Formula CIV 23wherein R.sup.3 is halo,
trifluoromethyl, cyano, (C.sub.1-C.sub.3)alkyl,
(C.sub.1-C.sub.3)alkyloxy- , ethenyl or ethynyl.
38. A compound as recited in claim 37 wherein R.sup.3 is
trifluoromethyl.
39. A compound as recited in claim 37 wherein R.sup.3 is
fluoro.
40-48. (cancelled)
49. A method of making a compound of Formula CVII 24wherein T is
(C.sub.1-C.sub.4)alkyl; R.sup.2 is H, (C.sub.1-C.sub.3)alkyl or
(C.sub.3-C.sub.5)cycloalkyl; and R.sup.3 is halo, trifluoromethyl,
cyano, (C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkyloxy, ethenyl
or ethynyl; comprising acylating a (C.sub.1-C.sub.4)alkylamine with
a Formula CVI compound 25wherein R.sup.2 is H,
(C.sub.1-C.sub.3)alkyl or (C.sub.3-C.sub.5)cycloalkyl; and wherein
R.sup.3 is halo, trifluoromethyl, cyano, (C.sub.1-C.sub.3)alkyl,
(C.sub.1-C.sub.3)alkyloxy- , ethenyl or ethynyl.
50. A method as recited in claim 49 wherein R.sup.2 is H or
cyclopropyl; R.sup.3 is fluoro, chloro or trifluoromethyl; and the
Formula CVI acid is esterified to a (C.sub.1-C.sub.6)alkyl ester
prior to acylation with the (C.sub.1-C.sub.4)alkylamine.
51. A method as recited in claim 50 wherein the Formula CVI acid is
esterified with an alcohol in the presence of acid at a temperature
of ambient to reflux for a period of about 1 hours to about 12
hours.
52. A method as recited in claim 51 wherein the ester is reacted
with the amine at a temperature of about ambient to reflux for
about one to about 12 hours in an alcohol solvent.
53. A method as recited in claim 52 wherein the esterification
occurs at a temperature of about 50.degree. c. and the acylation
occurs at a temperature of about 50.degree. c.
54. A method as recited in claim 53 wherein the alcohol is
methanol; the acid is HCl; the amine is methylamine; R.sup.2 is H;
and R.sup.3 is chloro.
55. A method as recited in claim 53 wherein the alcohol is
methanol; the acid is HCl; the amine is methylamine; R.sup.2 is
cyclopropyl; and R.sup.3 is fluoro.
56. A method as recited in claim 53 wherein the alcohol is
methanol; the acid is HCl; the amine is methylamine; R.sup.2 is H;
and R.sup.3 is trifluoromethyl.
57. A method of reducing tissue damage resulting from ischemia or
hypoxia comprising administering to a mammal in need of such
treatment a therapeutically effective amount of a compound of claim
1 or a prodrug thereof or a pharmaceutically acceptable salt of
said compound or of said prodrug.
58. A method as recited in claim 57 wherein the tissue is cardiac,
brain, liver, kidney, lung, gut, skeletal muscle, spleen, pancreas,
nerve, spinal cord, retina tissue, the vasculature, or intestinal
tissue.
59. A method as recited in claim 57 wherein the amount of the
Formula I compound is about 0.01 mg/kg/day to about 50
mg/kg/day.
60. A method as recited in claim 59 wherein the mammal is a female
or male human.
61. A method as recited in claim 60 wherein said tissue is heart
tissue.
62. A method as recited in claim 60 wherein said tissue is brain
tissue.
63. A method as recited in claim 60 wherein said tissue is liver
tissue.
64. A method as recited in claim 60 wherein said tissue is kidney
tissue.
65. A method as recited in claim 60 wherein said tissue is lung
tissue.
66. A method as recited in claim 60 wherein said tissue is gut
tissue.
67. A method as recited in claim 60 wherein said tissue is skeletal
muscle tissue.
68. A method as recited in claim 60 wherein said tissue is spleen
tissue.
69. A method as recited in claim 60 wherein said tissue is pancreas
tissue.
70. A method as recited in claim 60 wherein said tissue is retina
tissue.
71. A method as recited in claim 60 wherein the compound is
administered prophylactically.
72. A method as recited in claim 60 wherein the compound is
administered prior to surgery.
73. A method as recited in claim 60 wherein the compound is
administered prior to cardiac surgery.
74. A method as recited in claim 60 wherein the compound is
administered prior to, during and after surgery.
75. A method as recited in claim 60 wherein the compound is
administered prior to, during and after cardiac surgery.
76. A method as recited in claim 60 wherein the compound is
administered within twenty-four hours after surgery.
77. A method as recited in claim 60 wherein the compound is
administered within twenty four hours after cardiac surgery.
78. A method as recited in claim 60 wherein the tissue damage
resulting from ischemia or hypoxia is ischemic or hypoxic damage
and is incurred during organ transplantation.
79. A method as recited in claim 60 wherein the compound is
administered to prevent perioperative myocardial ischemic
injury.
80. A pharmaceutical composition which comprises a therapeutically
effective amount of a compound of claim 1 or a prodrug thereof or a
pharmaceutically acceptable salt of said compound or of said
prodrug and a pharmaceutically acceptable carrier, vehicle or
diluent.
81. A pharmaceutical composition for the reduction of tissue damage
resulting from ischemia or hypoxia which comprises a
therapeutically effective amount of a compound of claim 1 or a
prodrug thereof or a pharmaceutically acceptable salt of said
compound or of said prodrug and a pharmaceutically acceptable
carrier, vehicle or diluent.
82. A pharmaceutical combination composition comprising: a
therapeutically effective amount of a composition comprising a
first compound, said first compound being a compound of claim 1, a
prodrug thereof, or a pharmaceutically acceptable salt of said
compound or of said prodrug; a second compound, said second
compound being an aldose reductase inhibitor; and a pharmaceutical
carrier, vehicle or diluent.
83. A pharmaceutical composition as recited in claim 82 wherein the
aldose reductase inhibitor is 1-phthalazineacetic acid,
3,4-dihydro-4-oxo-3-[[5--
trifluoromethyl)-2-benzothiazolyl]methyl]- or a pharmaceutically
acceptable salt thereof.
84. A method of reducing tissue damage resulting from ischemia or
hypoxia comprising administering to a mammal in need of such
treatment an amount of a first compound, said first compound being
a compound of claim 1, a prodrug thereof, or a pharmaceutically
acceptable salt of said compound or of said prodrug; an amount of a
second compound, said second compound being an aldose reductase
inhibitor; wherein the amounts of the first and second compounds
result in a therapeutic effect.
85. A method of reducing tissue damage resulting from ischemia or
hypoxia as recited in claim 84 wherein the aldose reductase
inhibitor is 1-phthalazineacetic acid,
3,4-dihydro-4-oxo-3-[[5-trifluoromethyl)-2-benz-
othiazolyl]methyl]- or a pharmaceutically acceptable salt
thereof.
86. A kit comprising: a first compound, said first compound being a
compound of claim 1, a prodrug thereof, or a pharmaceutically
acceptable salt of said compound or of said prodrug and a
pharmaceutically acceptable carrier, vehicle or diluent in a first
unit dosage form; b. a second compound, said second compound being
an aldose reductase inhibitor and a pharmaceutically acceptable
carrier, vehicle or diluent in a second unit dosage form; and c.
means for containing said first and second dosage forms wherein the
amounts of first and second compounds result in a therapeutic
effect.
87. A kit as recited in claim 86 wherein the aldose reductase
inhibitor is 1-phthalazineacetic acid,
3,4-dihydro-4-oxo-3-[[5-trifluoromethyl)-2-benz-
othiazolyl]methyl]- or a pharmaceutically acceptable salt
thereof.
88. A pharmaceutical combination composition comprising: a
therapeutically effective amount of a composition comprising a
first compound, said first compound being a compound of claim 1, a
prodrug thereof, or a pharmaceutically acceptable salt of said
compound or of said prodrug; a second compound, said second
compound being a glycogen phosphorylase inhibitor; and a
pharmaceutical carrier, vehicle or diluent.
89. A pharmaceutical composition as recited in claim 88 wherein the
glycogen phosphorylase inhibitor is 5-chloro-1H-indole-2-carboxylic
acid
[(IS)-benzyl-(2R)-hydroxy-3-((3S)-hydroxypyrrolidin-1-yl)-3-oxopropyl]-am-
ide; 5-chloro-1H-indole-2-carboxylic acid
[(IS)-benzyl-3-((3S,4S)-dihydrox-
ypyrrolidin-1-yl)-(2R)-hydroxy-3-oxopropyl]-amide;
5-chloro-1-1H-indole-2-- carboxylic acid
[(1S)-((R)-hydroxy-dimethylcarbamoyl-methyl)-2-phenyl-ethy-
l]-amide; 5-chloro-1H-indole-2-carboxylic acid
[(1S)-((R)-hydroxy-methoxy--
methyl-carbamoyl)-methyl)-2-phenyl-ethyl]-amide;
5-chloro-1H-indole-2-carb- oxylic acid
[(1S)-((R)-hydroxy-[(2-hydroxy-ethyl)-methyl-carbamoyl]-methyl-
)-2-phenyl-ethyl]-amide; 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-(3-hydroxylamino-pyrrolidin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[2-(cis-3,4-dihydroxy-pyrrolidin-1-y- l)-2-oxo-ethyl!]-amide;
5-chloro-1H-indole-2-carboxylic acid [(IS)-benzyl
-3-((cis)-dihydroxypyrrolidin-1-yl)
-(2R)-hydroxy-3-oxopropyl]-amide; 5-chloro-1H-indole-2-carboxylic
acid [2-((3S,4S) -dihydroxy-pyrrolidin-1-- yl)-2-oxo-ethyl]-amide;
5-chloro-1 H-indole-2-carboxylic acid
[(1S)-benzyl-2-(cis-3,4-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[2-(1,1-dioxo-thiazolidin-3-yl)-2-ox- o-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid [(1
S)-(4-fluoro-benzyl)-2-(4-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid [(1
S)-benzyl-2-((3RS)-hydroxy-piper- idin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid [2-oxo-2-((1
RS)-oxo-thiazolidin-3-yl)-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[(S1)-benzyl-2-(3-hydroxy-azetidin-1- -yl)-2-oxo-ethyl]-amide; or a
pharmaceutically acceptable salt thereof.
90. A method of reducing tissue damage resulting from ischemia or
hypoxia comprising administering to a mammal in need of such
treatment an amount of a first compound, said first compound being
a compound of claim 1, a prodrug thereof, or a pharmaceutically
acceptable salt of said compound or of said prodrug; and an amount
of a second compound, said second compound being a glycogen
phosphorylase inhibitor; wherein the amounts of first and second
compounds result in a therapeutic effect.
91. A method of reducing tissue damage resulting from ischemia as
recited in claim 90 wherein the glycogen phosphorylase inhibitor is
5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-(2R)-hydroxy-3-((3S)-hy-
droxypyrrolidin-1-yl)-3-oxopropyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[(IS)-benzyl-3-((3S,4S)-dihydroxypyrrolidin-1-yl)-(2R)-hydroxy-3-oxo-
propyl]-amide; 5-chloro-1H-indole-2-carboxylic acid
[(1S)-((R)-hydroxy-dimethylcarbamoyl-methyl)-2-phenyl-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[(IS)-((R)-hydroxy-methoxy-methyl-ca-
rbamoyl)-methyl)-2-phenyl-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[(1S)-((R)-hydroxy-[(2-hydroxy-ethyl)-methyl-carbamoyl]-methyl)-2-ph-
enyl-ethyl]-amide; 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-(3-hydroxylamino-pyrrolidin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[2-(cis-3,4-dihydroxy-pyrrolidin-1-y- l)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid [(IS)-benzyl
-3-((cis)-dihydroxypyrrolidin-1yl)-(2R)-hydroxy-3-oxopropyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[2-((3S,4S)-dihydroxy-pyrrolidin-1-y- l)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid [(1
S)-benzyl-2-(cis-3,4-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[2-(1,1-dioxo-thiazolidin-3-yl)-2-ox- o-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid [(1
S)-(4-fluoro-benzyl)-2-(4-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid [(1
S)-benzyl-2-((3RS)-hydroxy-piper- idin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid [2-oxo-2-(( 1
RS)-oxo-thiazolidin-3-yl)-ethyl]-amide; 5-chloro-1
H-indole-2-carboxylic acid [(1S)-benzyl-2-(3-hydroxy
azetidin-1-yl)-2-oxo-ethyl]-amide; or a pharmaceutically acceptable
salt thereof.
92. A kit comprising: a. a first compound, said first compound
being a compound of claim 1, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug and a pharmaceutically acceptable carrier, vehicle or
diluent in a first unit dosage form; b. a second compound, said
second compound being an glycogen phosphorylase inhibitor and a
pharmaceutically acceptable carrier, vehicle or diluent in a second
unit dosage form; and c. means for containing said first and second
dosage forms wherein the amounts of first and second compounds
result in a therapeutic effect.
93. A kit as recited in claim 92 wherein the glycogen phosphorylase
inhibitor is 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-(2R)-hydro-
xy-3-((3S)-hydroxypyrrolidin-1-yl)-3-oxopropyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[(IS)-benzyl-3-((3S,4S)-dihydroxypyr-
rolidin-1-yl)-(2R)-hydroxy-3-oxopropyl]-amide;
5-chloro-1H-indole-2-carbox- ylic acid
[(1S)-((R)-hydroxy-dimethylcarbamoyl-methyl)-2-phenyl-ethyl]-ami-
de; -chloro-1H-indole-2-carboxylic acid
[(1S)-((R)-hydroxy-methoxy-methyl--
carbamoyl)-methyl)-2-phenyl-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[(1S)-((R)-hydroxy-[(2-hydroxy-ethyl)-methyl-carbamoyl]-methyl)-2-ph-
enyl-ethyl]-amide; 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-(3-hydroxylamino-pyrrolidin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[2-(cis-3,4-dihydroxy-pyrrolidin-1-y- l)-2oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid [(1S)-benzyl
-3-((cis)-dihydroxypyrrolidin-1-yl)-(2R)-hydroxy-3-oxopropyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[2-((3S,4S)-dihydroxy-pyrrolidin-1-y- l)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-(cis-3,4-dihydroxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[2-(1,1-dioxo-thiazolidin-3-yl)-2-ox- o-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[(1S)-(4-fluoro-benzyl)-2-(4-hydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1 H-indole-2-carboxylic acid
[(1S)-benzyl-2-((3RS)-hydroxy-piper- idin-1-yl)-2-oxo-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[2-oxo-2-((1RS)-oxo-thiazolidin-3-yl)-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[2-oxo-2-((1RS)-oxo-thiazolidin-3-yl- )-ethyl]-amide;
5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-(3-hydroxy-azetidin-1-yl)-2-oxo-ethyl]-amide; or a
pharmaceutically acceptable salt thereof.
94. A pharmaceutical combination composition comprising: a
therapeutically effective amount of a composition comprising a
first compound, said first compound being a compound of claim 1, a
prodrug thereof, or a pharmaceutically acceptable salt of said
compound or of said prodrug; a second compound, said second
compound being a cardiovascular agent; and a pharmaceutical
carrier, vehicle or diluent.
95. A pharmaceutical composition as recited in claim 94 wherein the
cardiovascular agent is a p-blocker, a calcium channel blocker, a
potassium channel opener, adenosine, adenosine agonists, an ACE
inhibitor, a nitrate, a diuretic, a glycoside, a thrombolytic, a
platelet inhibitor, aspirin, dipyridamol, potassium chloride,
clonidine, prazosin, pyruvate dehydrogenase kinase inhibitors,
pyruvate dehydrogenase complex activators, biguanides, NHE-1
inhibitor, Angiotensin II (All) receptor antagonists, C5a
inhibitors, soluble complement receptor type 1 (sCR1) or analogues,
partial fatty acid oxidation (PFOX) inhibitors (specifically,
ranolazine), acetyl CoA carboxylase activators, malonyl CoA
decarboxylase inhibitors, 5'AMP-activated protein kinase (AMPK)
inhibitors, adenosine nucleoside inhibitors, anti-apoptotic agents
(e.g., caspase inhibitors), monophosphoryl lipid A or analogues,
nitric oxide synthase activators/inhibitors, protein kinase C
activators (specifically, protein kinase .epsilon.), poly (ADP
ribose) synthetase (PARS, PARP) inhibitors, metformin
(gluconegenesis inhibitors, insulin sensitizers), endothelin
coverting enzyme (ECE) inhibitors, endothelin ETA receptor
antagonists, TAFI inhibitors, or a Na/Ca exchanger modulators.
96. A pharmaceutical composition as recited in claim 95 wherein the
NHE-1 inhibitor is
[1-(8-bromoquinolin-5-yl)-5-cyclopropyl-1H-pyrazole-4-carbon-
yl]guanidine;
[1-(6-chloroquinolin-5-yl)-5-cyclopropyl-1H-pyrazole-4-carbo-
nyl]guanidine;
[1-(indazol-7-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guan- idine;
[1-(benzimidazol-5-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidi-
ne;
[1-(1-isoquinolyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[5-cydopropyl-1-(4-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[5-cyclopropyl-1-(quinolin-5-yl)-1H-pyrazole-4-carbonyl]guanidine;
[5-cyclopropyl-1-(quinolin-8-yl)-1H-pyrazole-4-carbonyl]guanidine;
[1-(indazol-6-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(indazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(benzimidazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(1-methylbenzimidazol-6-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
1-(5-quinolinyl)-5-n-propyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(5-quinolinyl)-5-isopropyl-1H-pyrazole-4-carbonyl]guanidine;
[5-ethyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-methylbenzimidazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(1,4-benzodioxan-6-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(benzotriazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(3-chloroindazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(5-quinolinyl)-5-butyl-1H-pyrazole-4-carbonyl]guanidine;
[5-propyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[5-isopropyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-chloro-4-methylsulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl-
]guanidine;
[1-(2-chlorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guani-
dine;
[1-(2-trifluoromethyl-4-fluorophenyl)-5-cyclopropyl-1H-pyrazole-4-ca-
rbonyl]guanidine;
[1-(2-bromophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]-
guanidine;
[1-(2-fluorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanid-
ine;
[1-(2-chloro-5-methoxyphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]gu-
anidine;
[1-(2-chloro-4-methylaminosulfonylphenyl)-5-cyclopropyl-1H-pyrazo-
le-4-carbonyl]guanidine;
[1-(2,5-dichlorophenyl)-5-cyclopropyl-1H-pyrazole-
-4-carbonyl]guanidine;
[1-(2,3-dichlorophenyl)-5-cyclopropyl-1H-pyrazole-4-
-carbonyl]guanidine;
[1-(2-chloro-5-aminocarbonylphenyl)-5-cyclopropyl-1H--
pyrazole-4-carbonyljguanidine;
[1-(2-chloro-5-aminosulfonylphenyl)-5-cyclo-
propyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-fluoro-6-trifluoromethylphe-
nyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-chloro-5-methylsulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl-
]guanidine;
[1-(2-chloro-5-dimethylaminosulfonylphenyl)-5-cyclopropyl-1H-p-
yrazole-4-carbonyl]guanidine;
[1-(2-trifluoromethyl-4-chlorophenyl)-5-cycl-
opropyl-1H-pyrazole-4-carbonyl]guanidine;
[1-(2-chlorophenyl)-5-methyl-1H-- pyrazole-4-carbonyl]guanidine;
[5-methyl-1-(2-trifluoromethylphenyl)-1H-py-
razole-4-carbonyl]guanidine;
[5-ethyl-1-phenyl-1H-pyrazole-4-carbonyl]guan- idine;
[5-cyclopropyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carbonyl]g-
uanidine; [5-cyclopropyl-1-phenyl-1H-pyrazole-4-carbonyl]guanidine;
[5-cyclopropyl-1-(2,6-dichlorophenyl)-1H-pyrazole-4-carbonyl]guanidine
or the pharmaceutically acceptable salts of said compounds.
97. A method of reducing tissue damage resulting from ischemia or
hypoxia comprising administering to a mammal in need of such
treatment an amount of a first compound, said first compound being
a compound of claim 1, a prodrug thereof, or a pharmaceutically
acceptable salt of said compound or of said prodrug; an amount of a
second compound, said second compound being a cardiovascular agent;
wherein the amounts the of first and second compounds result in a
therapeutic effect.
98. A method of reducing tissue damage resulting from ischemia or
hypoxia as recited in claim 97 wherein the cardiovascular agent is
a (3-blocker, a potassium channel opener, adenosine, adenosine
agonists, a calcium channel blocker, an ACE inhibitor, a nitrate, a
diuretic, a glycoside, a chrombolytic, a platelet inhibitor,
aspirin, dipyridamol, potassium chloride, clonidine, prazosin,
pyruvate dehydrogenase kinase inhibitors, pyruvate dehydrogenase
complex activators, biguanides, NHE-1 inhibitor, Angiotensin II
(All) receptor antagonists, C5a inhibitors, soluble complement
receptor type 1 (sCR 1) or analogues, partial fatty acid oxidation
(PFOX) inhibitors (specifically, ranolazine), acetyl CoA
carboxylase activators, malonyl CoA decarboxylase inhibitors,
5'AMP-activated protein kinase (AMPK) inhibitors, adenosine
nucleoside inhibitors, anti-apoptotic agents (e.g., caspase
inhibitors), monophosphoryl lipid A or analogues, nitric oxide
synthase activators/inhibitors, protein kinase C activators
(specifically, protein kinase .epsilon.), poly (ADP ribose)
synthetase (PARS, PARP) inhibitors, metformin (gluconegenesis
inhibitors, insulin sensitizers), endothelin coverting enzyme (ECE)
inhibitors, endothelin ET A receptor antagonists, TAFI inhibitors,
or a Na/Ca exchanger modulators.
99. A kit comprising: a. a first compound, said first compound
being a compound of claim 1, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug and a pharmaceutically acceptable carrier, vehicle or
diluent in a first unit dosage form; b. a second compound, said
second compound being a cardiovascular agent and a pharmaceutically
acceptable carrier, vehicle or diluent in a second unit dosage
form; and c. means for containing said first and second dosage
forms wherein the amounts of the first and second compounds result
in a therapeutic effect.
100. A kit as recited in claim 99 wherein the cardiovascular agent
is a P-blocker, a calcium channel blocker, an ACE inhibitor, a
nitrate, a diuretic, a glycoside, a thrombolytic, a platelet
inhibitor, aspirin, dipyridamol, potassium chloride, clonidine,
prazosin, pyruvate dehydrogenase kinase inhibitors, pyruvate
dehydrogenase complex activators, biguanides or an NHE-1
inhibitor.
101. A compound selected from the group consisting of
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzyloxy-5-chloro-benzylamino)-purin-9-yl]--
4-hydroxytetrahydrofuran-2-carboxylic acid methylamide,
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(furan-3-ylmethoxy)benzylamino]-pur-
in-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid methylamide,
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(furan-2-ylmethoxy)benzylamino]puri-
n-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid methylamide,
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(thiazol-2-ylmethoxy)-benzylamino]--
purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylicacid
methylamide,
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(3-methylisoxazol-5-ylmethoxy)
benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide or the pharmaceutically acceptable salts of said
compounds.
Description
[0001] This application claims priority from provision application
U.S. Ser. No. 60/156,828 filed Sep. 30, 1999, the benefit of which
is hereby claimed under 37 C.F.R..sctn.1.78(a)(3).
BACKGROUND OF INVENTION
[0002] This invention relates to adenosine A-3 receptor agonists,
pharmaceutical compositions containing such inhibitors and the use
of such inhibitors to treat for example, ischemia particularly,
perioperative myocardial ischemic injury in mammals, including
humans.
[0003] Mycardial ischemic injury can occur in out-patient as well
as in perioperative settings and can lead to the development of
sudden death, myocardial infarction or congestive heart failure.
There is an unmet medical need to prevent or minimize myocardial
ischemic injury, particularly perioperative myocardial infarction.
Such a therapy is anticipated to be life-saving and reduce
hospitalizations, enhance quality of life and reduce overall health
care costs of high risk patients.
[0004] Pharmacological cardioprotection would reduce the incidence
and progression of myocardial infarction and dysfunction occurring
in these surgical settings (perioperatively). In addition to
reducing myocardial damage and improving post-ischemic myocardial
function in patients with ischemic heart disease, cardioprotection
would also decrease the incidence of cardiac morbidity and
mortality due to myocardial infarction and dysfunction in patients
"at risk" (such as greater than 65 years, exercise intolerant,
coronary artery disease, diabetes mellitus, hypertension) that
require non-cardiac surgery.
[0005] U.S. Pat. No. 5,604,210 discloses the use of certain
adenosine type compounds for the prevention or treatment of a brain
edema, an intracranial hemorrhage and a cerebral infarction.
[0006] U.S. Pat. No. 5,688,774 discloses A.sub.3 selective
agonists, particularly, adenine compounds having selected
substituents at the 2, 6 and 9 positions, and related substituted
compounds, particularly those containing substituents on the benzyl
and/or uronamide groups as agents which activate the A.sub.3
receptor.
[0007] U.S. Pat. No. 5,773,423 discloses
N.sup.6-benzyladenosine-5'-N-uron- amide and related substituted
compounds, particularly those containing substituents on the benzyl
and/or uronamide groups, and modified xanthine ribosides for the
activation of the A.sub.3 adenosine receptor.
[0008] J. Med. Chem. 1994, 37, 636-646, "Structure-Activity
Relationships of N.sup.6-Benzyladenosine-5'-uronamides as
A.sub.3-Selective Agonists" discloses the synthesis of adenosine
analogues modified at the 5'-position as uronamides and/or as
N.sup.6-benzyl derivatives which are potentially useful as
pharmacological and biochemical probes for A.sub.3 receptors.
[0009] J. Med. Chem. 1995, 38, 1174-1188, "Search for New Purine-
and Ribose-Modified Adenosine Analogues as Selective Agonists and
Antagonists at Adenosine Receptors", discloses that the binding
affinities at rat A.sub.1, A.sub.2, and A.sub.3 adenosine receptors
of a wide range of derivatives of adenosine have been determined.
In particular, 3'-.beta.-amino compounds were found to have no
activity.
[0010] J. Med. Chem. 1995, 38, 1720-1735, "Structure-Activity
Relationships of 9-Alkyladenine and Ribose-Modified Adenosine
Derivatives at Rat A.sub.3 Adenosine Receptors" discloses the
synthesis of 9-alkyladenine derivatives and ribose-modified
N.sup.6-benzyladenosine derivatives as leads for the development of
antagonists for the rat A.sub.3 adenosine receptor.
[0011] U.S. Pat. No. 5,817,760 discloses recombinant human
adenosine receptors A1, A2a, A2b, and A3 which were prepared by
cDNA cloning and polymerase chain reaction techniques. The
recombinant adenosine receptors can be utilized in an assay to
identify and evaluate entities that bind to or enhance binding to
adenosine receptors.
[0012] Thus, while there has been some progress in this field of
art, there is clearly a need and a continuing search in this field
of art for treatments for perioperative myocardial ischemia.
SUMMARY OF THE INVENTION
[0013] This invention is directed to compounds of Formula I 1
[0014] prodrugs thereof and pharmaceutically acceptable salts of
said compounds and of said prodrug, wherein
[0015] X is oxy, methylene or thio;
[0016] Y is CH or N;
[0017] Z is H, (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkyloxy,
trifluoromethyl or halo;
[0018] R.sup.1 is hydroxymethyl, (C.sub.1-C.sub.3)alkoxymethyl,
(C.sub.3-C.sub.5)cycloalkoxymethyl, carboxy,
(C.sub.1-C.sub.3)alkoxycarbo- nyl,
(C.sub.3-C.sub.5)cycloalkoxycarbonyl, 1,1-aminoiminomethyl,
1,1-(mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylamino)iminomethyl, 1,1
-(mono-N- or di-N,N-(C.sub.3-C.sub.5)cycloalkylamino)iminomethyl,
carbamoyl, mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl,
mono-N- or di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl or
N-(C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl;
[0019] R.sup.2 is H, (C.sub.1-C.sub.3)alkyl or
(C.sub.3-C.sub.5)cycloalkyl- ;
[0020] R.sup.3 is halo, trifluoromethyl, cyano,
(C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkyloxy, ethenyl or
ethynyl;
[0021] D is oxy, thio, NH, (C.sub.1-C.sub.6)alkyloxy,
(C.sub.1-C.sub.6)alkylthio or (C.sub.1-C.sub.6)alkylamino;
[0022] G is a partially saturated, fully saturated or fully
unsaturated five to eight membered ring optionally having one to
three heteroatoms selected independently from oxygen, sulfur and
nitrogen, or a bicyclic ring consisting of two fused partially
saturated, fully saturated or fully unsaturated three to six
membered rings, taken independently, optionally having one to four
heteroatoms selected independently from nitrogen, sulfur and
oxygen; wherein said G is optionally mono-, di- or tri-substituted
independently with halo, (C.sub.1-C.sub.3)alkyl, trifluoromethyl,
trifluoromethoxy, nitro, cyano, (C.sub.3-C.sub.5)cycloal- kyl,
hydroxy or (C.sub.1-C.sub.3)alkoxy or
[0023] G is cyano, (C.sub.1-C.sub.4)alkoxycarbonyl,
(C.sub.3-C.sub.5)cycloalkoxycarbonyl, C(O)NR.sup.4R.sup.5,
C(S)NR.sup.4R.sup.5, C(NH)NR.sup.4R.sup.5,
C(N(C.sub.1-C.sub.3)alkyl)NR.s- up.4R.sup.5 or
C(N(C.sub.3-C.sub.10) cycloalkyl)NR.sup.4R.sup.5,
[0024] R.sup.4 is a bond, H, (C.sub.1-C.sub.10)alkyl, hydroxy,
(C.sub.1-C.sub.10)alkoxy, (C.sub.3-C.sub.10)cycloalkoxy or a
partially saturated, fully saturated or fully unsaturated five to
eight membered ring, optionally linked through
(C.sub.1-C.sub.3)alkyl, optionally having one to three heteroatoms
selected independently from oxygen, sulfur and nitrogen, or, a
bicyclic ring or a bicyclic ring with optional (C.sub.1-C.sub.3)
bridge (e.g., adamantane) optionally linked through
(C.sub.1-C.sub.3)alkyl, said bicyclic ring or bridged bicyclic ring
optionally having one to four heteroatoms selected independently
from nitrogen, sulfur and oxygen wherein said
(C.sub.1-C.sub.10)alkyl,(C.sub.1- -C.sub.10)alkoxy,
(C.sub.3-C.sub.10)cycloalkoxy or R.sup.4 ring(s) is optionally
mono-, di- or tri-substituted independently with halo,
(C.sub.1-C.sub.3)alkyl, trifluoromethyl, nitro, cyano,
(C.sub.3-C.sub.5)cycloalkyl, hydroxy or
(C.sub.1-C.sub.3)alkoxy;
[0025] R.sup.5 is a bond, H, (C.sub.1-C.sub.10)alkyl or
(C.sub.1-C.sub.10)cycloalkyl; or
[0026] R.sup.4 and R.sup.5 taken together with the nitrogen to
which they are attached form a fully saturated or partially
unsaturated four to nine membered ring, said ring optionally
bridged, optionally having one to three additional heteroatoms
selected independently from oxygen, sulfur and nitrogen, said ring
optionally mono- or di-substituted independently with oxo, hydroxy,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.8)alkyl, amino, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
N-(C.sub.1-C.sub.4)alkyl-
-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cy- cloalkylamino,
N-(C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylamino- ,
formylamino, (C.sub.1-C.sub.4)alkylcarbonylamino,
(C.sub.3-C.sub.5)cycloalkylcarbonylamino,
(C.sub.1-C.sub.4)alkoxycarbonyl- amino,
N-(C.sub.1-C.sub.4)alkoxycarbonyl-N-(C.sub.1-C.sub.4)alkylamino,
(C.sub.1-C.sub.4)sulfamoyl, (C.sub.1-C.sub.4)alkylsulfonylamino,
(C.sub.3-C.sub.5)cycloalkylsulfonylamino or a partially saturated,
fully saturated or fully unsaturated five to eight membered ring,
optionally linked through (C.sub.1-C.sub.3)alkyl, optionally having
one to three heteroatoms selected independently from oxygen, sulfur
and nitrogen, or a bicyclic ring consisting of two fused partially
saturated, fully saturated or fully unsaturated three to six
membered rings, taken independently, optionally linked through
(C.sub.1-C.sub.3)alkyl, optionally having one to four heteroatoms
selected independently from nitrogen, sulfur and oxygen, optionally
mono- or di-substituted with halo, trifluoromethyl,
trifluoromethoxy, (C.sub.1-C.sub.3)alkyl or
(C.sub.1-C.sub.3)alkoxy.
[0027] A preferred group of compounds, designated the A Group,
contains those compounds having the Formula I as shown above
wherein
[0028] X is oxy;
[0029] Y is N;
[0030] Z is H;
[0031] R.sup.1 is (C.sub.1-C.sub.6)alkylcarbamoyl;
[0032] R.sup.2 is H;
[0033] R.sup.3 is halo, trifluoromethyl, cyano,
(C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkyloxy, ethenyl or
ethynyl;
[0034] D is oxy, thio, (C.sub.1-C.sub.6)alkyloxy or
(C.sub.1-C.sub.6)alkylthio;
[0035] G is phenyl, pyridyl, pyrimidinyl, pyrazinyl, thiazolyl,
oxazolyl, isoxazolyl, pyridinazinyl, tetrazolyl, isothiazolyl,
thiophenyl, furanyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl,
pyrazolyl, pyrrolyl, indolyl, naphthalenyl, quinolinyl,
isoquinolinyl, benzo[b]furanyl, benzo[b]thiophenyl, benzothiazolyl,
tetrahydrofuranyl, pyrrolidinyl, piperidinyl, tetrahydropyranyl,
morpholinyl wherein said G is optionally mono-, di- or
tri-substituted independently with halo, (C.sub.1-C.sub.3)alkyl or
(C.sub.1-C.sub.3)alkoxy; and pharmaceutically acceptable salts
thereof.
[0036] A group of compounds which is preferred among the A Group of
compounds, designated the B Group, contains those compounds
wherein
[0037] R.sup.1 is methylcarbamoyl;
[0038] R.sup.3 is halo;
[0039] D is (C.sub.1-C.sub.6)alkoxy;
[0040] G is phenyl, pyridyl, thiazolyl, oxazolyl, isoxazolyl,
isothiazolyl, furanyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl,
pyrazolyl, pyrrolyl wherein said G is optionally mono-, di- or
tri-substituted independently with halo, (C.sub.1-C.sub.3)alkyl,
trifluoromethoxy or (C.sub.1-C.sub.3)alkoxy; and pharmaceutically
acceptable salts thereof.
[0041] A group of compounds which is preferred among the B Group of
compounds, designated the C Group, contains those compounds
wherein
[0042] D is (C.sub.1-C.sub.2)alkoxy;
[0043] G is phenyl, thiazolyl, oxazolyl, isoxazolyl,
tetrahydrofuranyl, pyrrolidinyl, piperidinyl or morpholinyl wherein
said G is optionally mono-, di- or tri-substituted independently
with halo, (C.sub.1-C.sub.3)alkyl or (C.sub.1-C.sub.3)alkoxy; and
pharmaceutically acceptable salts thereof.
[0044] Especially preferred compounds within the B Group of
compounds are compounds wherein
[0045] a. R.sup.3 is chloro;
[0046] D is methyleneoxy; and
[0047] G is phenyl,
[0048] b. R.sup.3 is chloro;
[0049] D is methyleneoxy; and
[0050] G is 3-furanyl,
[0051] c. R.sup.3 is chloro;
[0052] D is methyleneoxy; and
[0053] G is 2-furanyl,
[0054] d. R.sup.3 is chloro,
[0055] D is methyleneoxy; and
[0056] G is 2-thiazolyl,
[0057] e. R.sup.3 is chloro;
[0058] D is methyleneoxy; and
[0059] G is 5-(3-methylisoxazolyl); and the pharmaceuticallyl
acceptable salts of said compounds.
[0060] Especially preferred compounds of this invention are the
compounds
(2S,3S,4R,5R)3-amino-5-[6-(2-benzyloxy-5-chloro-benzylamino)-purin-9-yl]--
4-hydroxytetrahydrofuran-2-carboxylic acid methylamide,
(2S,3S,4R,5R)3-amino-5-{6-[5-chloro-2-(furan-3-ylmethoxy)benzylamino]-pur-
in-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid methylamide,
(2S,3S,4R,5R)3-amino-5-{6-[5-chloro-2-(furan-2-ylmethoxy)benzylamino]-pur-
in-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid methylamide,
(2S,3S,4R,5R)3-amino-5-{6-[5-chloro-2-(thiazol-2-ylmethoxy)-benzylamino]--
purin-9-yl}4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide,
(2S,3S,4R,5R)3-amino-5-{6-[5-chloro-2-(3-methylisoxazol-5-ylmethoxy)
benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide, and pharmaceutically acceptable salts of said
compounds.
[0061] A preferred group of compounds, designated the D Group,
contains those compounds having the Formula I as shown above
wherein
[0062] X is oxy;
[0063] Y is N;
[0064] Z is H;
[0065] R.sup.1 is (C.sub.1-C.sub.6)alkylcarbamoyl;
[0066] R.sup.2 is H;
[0067] R.sup.3 is halo, trifluoromethyl, cyano,
(C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkyloxy, ethenyl or
ethynyl;
[0068] D is (C.sub.1-C.sub.6)alkyloxy or
(C.sub.1-C.sub.6)alkylthio;
[0069] G is C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4R.sup.5
[0070] R.sup.4 and R.sup.5 taken together with the nitrogen to
which they are attached form a fully saturated four to nine
membered ring, optionally having one to three additional
heteroatoms selected independently from oxygen, sulfur and
nitrogen, said ring optionally mono- or di-substituted
independently with oxo, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.8)alkyl, amino, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylam- inocarbonyl, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
N-(C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylamino, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylamino or
N-(C.sub.1-C.sub.4)alkyl-N-(C.- sub.3-C.sub.5)cycloalkylamino,
formylamino, (C.sub.1-C.sub.4)alkylformylam- ino,
(C.sub.3-C.sub.5)cycloalkylformylamino, sulfamoyl,
(C.sub.1-C.sub.4)alkylsulfonylamino,
(C.sub.3-C.sub.5)cycloalkylsulfonyla- mino or a partially
saturated, fully saturated or fully unsaturated five to eight
membered ring, optionally linked through (C.sub.1-C.sub.3)alkyl,
optionally having one to three heteroatoms selected independently
from oxygen, sulfur and nitrogen, or, a bicyclic ring consisting of
two fused partially saturated, fully saturated or fully unsaturated
three to six membered rings, taken independently, optionally linked
through (C.sub.1-C.sub.3)alkyl, optionally having one to four
heteroatoms selected independently from nitrogen, sulfur and
oxygen; and pharmaceutically acceptable salts thereof.
[0071] A preferred group of compounds which is preferred among the
D Group of compounds, designated the E Group, contains those
compounds wherein
[0072] R.sup.1 is methylcarbamoyl;
[0073] R.sup.3 is halo;
[0074] D is (C.sub.1-C.sub.2)alkoxy;
[0075] G is C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4R.sup.5;
[0076] wherein R.sup.4 and R.sup.5 taken together with the nitrogen
to which they are attached form piperidinyl, piperazinyl,
morpholinyl, azetidinyl or pyrrolidinyl said ring optionally mono-
or di-substituted independently with oxo, hydroxy,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.8)alkyl, amino, carbamoyl,
mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
N-(C.sub.1-C.sub.4)alkyl-
-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cy- cloalkylamino or
N-(C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylami- no,
formylamino, (C.sub.1-C.sub.4)alkylformylamino,
(C.sub.3-C.sub.5)cycloalkylformylamino, sulfamoyl,
(C.sub.1-C.sub.4)alkylsulfonylamino,
(C.sub.3-C.sub.5)cycloalkylsulfonyla- mino or
[0077] a partially saturated, fully saturated or fully unsaturated
four to eight membered ring, optionally linked through
(C.sub.1-C.sub.3)alkyl, optionally having one to two heteroatoms
selected independently from oxygen, sulfur and nitrogen; and
pharmaceutically acceptable salts thereof.
[0078] A preferred group of compounds which is preferred among the
E Group of compounds, designated the F Group, contains those
compounds wherein
[0079] G is C(O)NR.sup.4R.sup.5;
[0080] wherein R.sup.4 and R.sup.5 taken together with the nitrogen
to which they are attached form piperidinyl, piperazinyl,
morpholinyl, azetidinyl, pyrrolidinyl said ring optionally mono- or
di-substituted independently with hydroxy,
oxo,(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.8)alkyl, amino,
carbamoyl, mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl,
mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylamino, or a partially
saturated, fully saturated or fully unsaturated four to eight
membered ring, optionally linked through (C.sub.1-C.sub.3)alkyl,
optionally having one to two heteroatoms selected independently
from oxygen, sulfur and nitrogen; and pharmaceutically acceptable
salts thereof.
[0081] Especially preferred compounds within the F Group of
compounds are compounds wherein
[0082] a. R.sup.3 is chloro;
[0083] D is methyleneoxy;
[0084] G is C(O)NR R;
[0085] wherein R.sup.4 and R.sup.5 taken together with the nitrogen
to which they are attached form piperazinyl substituted in the four
position with methyl,
[0086] b. R.sup.3 is chloro;
[0087] D is methyleneoxy;
[0088] G is C(O)NR.sup.4R.sup.5;
[0089] wherein R.sup.4 and R.sup.5 taken together with the nitrogen
to which they are attached form piperazinyl,
[0090] c. R.sup.3 is chloro;
[0091] D is methyleneoxy;
[0092] G is C(O)NR.sup.4R.sup.5;
[0093] wherein R.sup.4 and R.sup.5 taken together with the nitrogen
to which they are attached form piperidinyl substituted in the four
position with N,N-dimethylamino,
[0094] R.sup.3 is chloro;
[0095] D is methyleneoxy;
[0096] G is C(O)NR.sup.4R.sup.5;
[0097] wherein R.sup.4 and R.sup.5 taken together with the nitrogen
to which they are attached form piperidinyl substituted in the four
position with piperidin-1-yl,
[0098] e. R.sup.3 is chloro;
[0099] D is methyleneoxy;
[0100] G is C(O)NR.sup.4R.sup.5;
[0101] wherein R.sup.4 and R.sup.5 taken together with the nitrogen
to which they are attached form piperidinyl substituted in the four
position with methylamino, and pharmaceutically acceptable salts of
said compounds.
[0102] A preferred group of compounds, designated the G Group,
contains those compounds having the Formula I as shown above
wherein
[0103] X is oxy;
[0104] Y is N;
[0105] Z is H;
[0106] R.sup.1 is (C.sub.1-C.sub.6)alkylcarbamoyl;
[0107] R.sup.2 is H;
[0108] R.sup.3 is halo, trifluoromethyl, cyano,
(C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkyloxy, ethenyl or
ethynyl;
[0109] D is (C.sub.1-C.sub.6)alkyloxy or
(C.sub.1-C.sub.6)alkylthio;
[0110] G is C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4R.sup.5;
[0111] R.sup.4 is H, (C.sub.1-C.sub.10)alkyl, hydroxy,
(C.sub.1-C.sub.10)alkoxy, (C.sub.3-C.sub.10)cycloalkoxy or a
partially saturated, fully saturated or fully unsaturated five to
eight membered ring, optionally linked through
(C.sub.1-C.sub.3)alkyl, optionally having one to three heteroatoms
selected independently from oxygen, sulfur and nitrogen, or a
bicyclic ring consisting of two fused partially saturated, fully
saturated or fully unsaturated three to six membered rings, taken
independently, optionally linked through (C.sub.1-C.sub.3)alkyl,
optionally having one to four heteroatoms selected independently
from nitrogen, sulfur and oxygen;
[0112] R.sup.5 is H, (C.sub.1-C.sub.10)alkyl or
(C.sub.1-C.sub.10)cycloalk- yl; and pharmaceutically acceptable
salts thereof.
[0113] A group of compounds which is preferred among the G Group of
compounds, designated the H Group, contains those compounds
wherein
[0114] R.sup.1 is methylcarbamoyl;
[0115] R.sup.3 is halo;
[0116] D is (C.sub.1-C.sub.2)alkoxy;
[0117] G is C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4R.sup.5;
[0118] R.sup.4 is H, (C.sub.1-C.sub.10)alkyl, hydroxy,
(C.sub.1-C.sub.10)alkoxy, (C.sub.3-C.sub.10)cycloalkoxy or a
partially saturated, fully saturated or fully unsaturated five to
eight membered ring, optionally linked through
(C.sub.1-C.sub.3)alkyl, optionally having one to three heteroatoms
selected independently from oxygen, sulfur and nitrogen;
[0119] R.sup.5 is H, (C.sub.1-C.sub.10)alkyl or
(C.sub.1-C.sub.10)cycloalk- yl; and pharmaceutically acceptable
salts thereof.
[0120] A group of compounds which is preferred among the H Group of
compounds, designated the I Group, contains those compounds
wherein
[0121] G is C(O)NR.sup.4R.sup.5;
[0122] R.sup.4 is H, (C.sub.1-C.sub.10)alkyl,
(C.sub.3-C.sub.6)cycloalkyl, hydroxy, (C.sub.1-C.sub.10)alkoxy or
(C.sub.3-C.sub.10)cycloalkoxy;
[0123] R.sup.5 is H, (C.sub.1-C.sub.10)alkyl or
(C.sub.3-C.sub.10)cycloalk- yl; and pharmaceutically acceptable
salts thereof.
[0124] An especially preferred compound within the I Group of
compounds is the compound wherein
[0125] R.sup.3 is chloro;
[0126] D is methyleneoxy;
[0127] G is C(O)NR.sup.4R.sup.5;
[0128] R.sup.4 is H;
[0129] R.sup.5 is H; and pharmaceutically acceptable salts
thereof.
[0130] A preferred group of compounds, designated the J Group,
contains those compounds having the Formula I as shown above
wherein
[0131] D is oxy, thio, (C.sub.1-C.sub.6)alkyloxy or
(C.sub.1-C.sub.6)alkylthio;
[0132] G is phenyl, pyridyl, pyrimidinyl, pyrazinyl, thiazolyl,
oxazolyl, isoxazolyl, pyridinazinyl, tetrazolyl, isothiazolyl,
thiophenyl, furanyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl,
pyrazolyl, pyrrolyl, indolyl, naphthalenyl, quinolinyl,
isoquinolinyl, benzo[b]furanyl, benzo[b]thiophenyl, benzothiazolyl,
tetrahydrofuranyl, pyrrolidinyl, piperidinyl, tetrahydropyranyl,
morpholinyl wherein said G is optionally mono-, di- or
tri-substituted independently with halo, (C.sub.1-C.sub.3)alkyl or
(C.sub.1-C.sub.3)alkoxy; and pharmaceutically acceptable salts
thereof.
[0133] A group of compounds which is preferred among the J Group of
compounds, designated the K Group, contains those compounds
wherein
[0134] D is (C.sub.1-C.sub.6)alkoxy;
[0135] G is phenyl, pyridyl, thiazolyl, oxazolyl, isoxazolyl,
isothiazolyl, furanyl, 1,2,4-oxadiazolyl, 1,2,4-thiadiazolyl,
pyrazolyl, pyrrolyl wherein said G is optionally mono-, di- or
tri-substituted independently with halo, (C.sub.1-C.sub.3)alkyl or
(C.sub.1-C.sub.3)alkoxy; and pharmaceutically acceptable salts
thereof.
[0136] A preferred group of compounds, designated the L Group,
contains those compounds having the Formula I as shown above
wherein
[0137] D is (C.sub.1-C.sub.6)alkyloxy or
(C.sub.1-C.sub.6)alkylthio;
[0138] G is C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4R.sup.5 wherein
R.sup.4 and R.sup.5 taken together with the nitrogen to which they
are attached form a fully saturated four to nine membered ring,
optionally having one to three additional heteroatoms selected
independently from oxygen, sulfur and nitrogen, said ring
optionally mono- or di-substituted independently with oxo, hydroxy,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.8)alkyl, amino, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
N-(C.sub.1-C.sub.4)alkyl-
-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cy- cloalkylamino or
N-(C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylami- no,
formylamino, (C.sub.1-C.sub.4)alkylformylamino,
(C.sub.3-C.sub.5)cycloalkylformylamino, sulfamoyl,
(C.sub.1-C.sub.4)alkylsulfonylamino,
(C.sub.3-C.sub.5)cycloalkylsulfonyla- mino or a partially
saturated, fully saturated or fully unsaturated five to eight
membered ring, optionally linked through (C.sub.1-C.sub.3)alkyl,
optionally having one to three heteroatoms selected independently
from oxygen, sulfur and nitrogen, or a bicyclic ring consisting of
two fused partially saturated, fully saturated or fully unsaturated
three to six membered rings, taken independently, optionally linked
through (C.sub.1-C.sub.3)alkyl, optionally having one to four
heteroatoms selected independently from nitrogen, sulfur and
oxygen; and pharmaceutically acceptable salts thereof.
[0139] A group of compounds which is preferred among the L Group of
compounds, designated the M Group, contains those compounds
wherein
[0140] D is (C.sub.1-C.sub.2)alkoxy;
[0141] G is C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4R.sup.5;
[0142] wherein R.sup.4 and R.sup.5 taken together with the nitrogen
to which they are attached form piperidinyl, piperazinyl,
morpholinyl, azetidinyl or pyrrolidinyl said ring optionally mono-
or di-substituted independently with oxo, (C.sub.1-C.sub.6)alkoxy,
(C.sub.1-C.sub.8)alkyl, amino, carbamoyl, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
N-(C.sub.1-C.sub.4)alkyl-N-(C.sub.3-C.sub.5)cycloalkylaminocarbonyl,
mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylamino, mono-N- or
di-N,N-(C.sub.3-C.sub.5)cycloalkylamino or
N-(C.sub.1-C.sub.4)alkyl-N-(C.- sub.3-C.sub.5)cycloalkylamino,
formylamino, (C.sub.1-C.sub.4)alkylformylam- ino,
(C.sub.3-C.sub.5)cycloalkylformylamino, sulfamoyl,
(C.sub.1-C.sub.4)alkylsulfonylamino,
(C.sub.3-C.sub.5)cycloalkylsulfonyla- mino or a partially
saturated, fully saturated or fully unsaturated four to eight
membered ring, optionally linked through (C.sub.1-C.sub.3)alkyl,
optionally having one to two heteroatoms selected independently
from oxygen, sulfur and nitrogen; and pharmaceutically acceptable
salts thereof.
[0143] A group of compounds which is preferred among the M Group of
compounds, designated the N Group, contains those compounds
wherein
G is C(O)NR.sup.4R.sup.5;
[0144] wherein R.sup.4 and R.sup.5 taken together with the nitrogen
to which they are attached form piperidinyl, piperazinyl,
morpholinyl, azetidinyl, pyrrolidinyl said ring optionally mono- or
di-substituted independently with oxo, hydroxy,
(C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.8)alkyl, amino, carbamoyl,
mono-N- or di-N,N-(C.sub.1-C.sub.4)alkylaminocarbonyl, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino, or a partially saturated, fully
saturated or fully unsaturated four to eight membered ring,
optionally linked through (C.sub.1-C.sub.3)alkyl, optionally having
one to two heteroatoms selected independently from oxygen, sulfur
and nitrogen; and pharmaceutically acceptable salts thereof.
[0145] A preferred group of compounds, designated the O Group,
contains those compounds having the Formula I as shown above
wherein
[0146] D is (C.sub.1-C.sub.6)alkyloxy or
(C.sub.1-C.sub.6)alkylthio;
[0147] G is C(O)NR.sup.4R.sup.5 or C(S)NR.sup.4R.sup.5;
[0148] R.sup.4 is H, (C.sub.1-C.sub.10)alkyl, hydroxy,
(C.sub.1-C.sub.10)alkoxy, (C.sub.3-C.sub.10)cycloalkoxy or a
partially saturated, fully saturated or fully unsaturated five to
eight membered ring, optionally linked through
(C.sub.1-C.sub.3)alkyl, optionally having one to three heteroatoms
selected independently from oxygen, sulfur and nitrogen, or a
bicyclic ring consisting of two fused partially saturated, fully
saturated or fully unsaturated three to six membered rings, taken
independently, optionally linked through (C.sub.1-C.sub.3)alkyl,
optionally having one to four heteroatoms selected independently
from nitrogen, sulfur and oxygen;
[0149] R.sup.5 is H, (C.sub.1-C.sub.10)alkyl or
(C.sub.1-C.sub.10)cycloalk- yl;
[0150] and pharmaceutically acceptable salts thereof.
[0151] A group of compounds which is preferred among the O Group of
compounds, designated the P Group, contains those compounds
wherein
D is (C.sub.1-C.sub.2)alkoxy;
[0152] R.sup.4 is H, (C.sub.1-C.sub.10)alkyl, hydroxy,
(C.sub.1-C.sub.10)alkoxy, (C.sub.3-C.sub.10)cycloalkoxy or a
partially saturated, fully saturated or fully unsaturated five to
eight membered ring, optionally linked through
(C.sub.1-C.sub.3)alkyl, optionally having one to three heteroatoms
selected independently from oxygen, sulfur and nitrogen;
[0153] R.sup.5 is H, (C.sub.1-C.sub.10)alkyl or
(C.sub.1-C.sub.10)cycloalk- yl; and pharmaceutically acceptable
salts thereof.
[0154] A group of compounds which is preferred among the P Group of
compounds, designated the Q Group, contains those compounds
wherein
[0155] G is C(O)NR.sup.4R.sup.5;
[0156] R.sup.4 is H, (C.sub.1-C.sub.10)alkyl,
(C.sub.3-C.sub.6)cycloalkyl, hydroxy, (C.sub.1-C.sub.10)alkoxy or
(C.sub.3-C.sub.10)cycloalkoxy;
[0157] R.sup.5 is H, (C.sub.1-C.sub.10)alkyl or
(C.sub.3-C.sub.10)cycloalk- yl; and pharmaceutically acceptable
salts thereof.
[0158] Another aspect of this invention is directed to compounds
having the Formula C 2
[0159] wherein R.sup.20 and R.sup.21 are each independently
(C.sub.1-C.sub.4)alkyl, H, phenyl, phenyl(C.sub.1-C.sub.4)alkyl or
are joined together to form a piperidinyl, pyrrolidinyl or
morpholinyl ring;
[0160] R.sup.22 and R.sup.23 are each independently
(C.sub.1-C.sub.4)alkyl or are joined together to form a 5-6
membered carbocyclic ring; and
[0161] R.sup.24 is (C.sub.1-C.sub.4)alkyl, phenyl or
phenyl(C.sub.1-C.sub.4)alkyl, said phenyl or
phenyl(C.sub.1-C.sub.4)alkyl optionally mono-, di, or
tri-substituted independently on the phenyl moiety with nitro, halo
or trifluoromethyl.
[0162] Another aspect of this invention is directed to compounds
having the Formula CI 3
[0163] wherein R.sup.20 and R.sup.21 are each independently
(C.sub.1-C.sub.4)alkyl, H, phenyl, phenyl(C.sub.1-C.sub.4)alkyl or
are joined together to form a piperidinyl, pyrrolidinyl or
morpholinyl ring; and
[0164] R.sup.24 is (C.sub.1-C.sub.4)alkyl, phenyl or phenyl
(C.sub.1-C.sub.4)alkyl, said phenyl or phenyl(C.sub.1-C.sub.4)alkyl
optionally mono-, di, or tri-substituted independently on the
phenyl moiety with nitro, halo or trifluoromethyl.
[0165] Another aspect of this invention is directed to compounds
having the Formula CIII 4
[0166] wherein R.sup.20 and R.sup.21 are each independently
(C.sub.1-C.sub.4)alkyl, H, phenyl, phenyl(C.sub.1-C.sub.4)alkyl or
are joined together to form a piperidinyl, pyrrolidinyl or
morpholinyl ring;
[0167] R.sup.24 is (C.sub.1-C.sub.4)alkyl, phenyl or phenyl
(C.sub.1-C.sub.4)alkyl, said phenyl or phenyl(C.sub.1-C.sub.4)alkyl
optionally mono-, di, or tri-substituted independently on the
phenyl moiety with nitro, halo or trifluoromethyl; and
[0168] R.sup.25 and R.sup.26 are each independently
(C.sub.1-C.sub.4)alkyl or phenyl.
[0169] Another aspect of this invention is directed to compounds
having the Formula CIII 5
[0170] wherein R.sup.3 is halo, trifluoromethyl, cyano,
(C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkyloxy, ethenyl or
ethynyl.
[0171] Especially preferred compounds having Formula CIII as shown
above are compounds wherein
[0172] a. R.sup.3 is trifluoromethyl;
[0173] b. R.sup.3 is fluoro; and
[0174] c. R.sup.3 is chloro.
[0175] Another aspect of this invention is directed to compounds
having Formula CIV 6
[0176] wherein R.sup.3 is halo, trifluoromethyl, cyano,
(C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkyloxy, ethenyl or
ethynyl.
[0177] Especially preferred compounds having Formula CIV as shown
above are compounds wherein
[0178] a. R.sup.3 is trifluoromethyl;
[0179] b. R.sup.3 is fluoro; and
[0180] c. R.sup.3 is chloro.
[0181] Another aspect of this invention is directed to compounds
having the Formula CV 7
[0182] wherein R.sup.3 is halo, trifluoromethyl, cyano,
(C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkyloxy, ethenyl or
ethynyl.
[0183] Especially preferred compounds having Formula CV as shown
above are compounds wherein
[0184] a. R.sup.3 is trifluoromethyl;
[0185] b. R.sup.3 is fluoro; and
[0186] c. R.sup.3 is chloro.
[0187] Another aspect of this invention is directed to compounds
having the Formula CVI 8
[0188] wherein R.sup.2 is H, (C.sub.1-C.sub.3)alkyl or
(C.sub.3-C.sub.5)cycloalkyl; and
[0189] R.sup.3 is halo, trifluoromethyl, cyano,
(C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkyloxy, ethenyl or
ethynyl.
[0190] Especially preferred compounds having Formula CVI as shown
above are compounds wherein
[0191] a. R.sup.2 is H; and
[0192] R.sup.3 is chloro,
[0193] R.sup.2 is H; and
[0194] R.sup.3 is fluoro,
[0195] c. R.sup.2 is cyclopropyl; and
[0196] R.sup.3 is fluoro.
[0197] Another aspect of this invention is directed to a method of
making a compound of Formula CVII 9
[0198] wherein T is (C.sub.1-C.sub.4)alkyl;
[0199] R.sup.2 is H, (C.sub.1-C.sub.3)alkyl or
(C.sub.3-C.sub.5)cycloalkyl- ; and
[0200] R.sup.3 is halo, trifluoromethyl, cyano,
(C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkyloxy, ethenyl or
ethynyl; comprising acylating a (C.sub.1-C.sub.4alkyl)amine with a
Formula CVI compound. 10
[0201] wherein
[0202] R.sup.2 is H, (C.sub.1-C.sub.3)alkyl or
(C.sub.3-C.sub.5)cycloalkyl- ; and
[0203] R.sup.3 is halo, trifluoromethyl, cyano,
(C.sub.1-C.sub.3)alkyl, (C.sub.1-C.sub.3)alkyloxy, ethenyl or
ethynyl.
[0204] A preferred aspect of the above method is wherein
[0205] R.sup.2 is H or cyclopropyl;
[0206] R.sup.3 is fluoro, chloro or trifluoromethyl;
[0207] and the Formula CVI acid is esterified to a
(C.sub.1-C.sub.6)alkyl ester prior to acylation with the
(C.sub.1-C.sub.4)alkylamine.
[0208] An especially preferred aspect of the immediately preceding
method is wherein the Formula CVI acid is esterified with an
alcohol in the presence of acid at a temperature of ambient to
reflux for a period of about 1 hour to about 12 hours.
[0209] An especially preferred aspect of the immediately preceding
method, is wherein the ester is reacted with the amine at a
temperature of about ambient to reflux for about one to about 12
hours in an alcohol solvent.
[0210] An especially preferred aspect of the immediately preceding
method designated the X method is, wherein the esterification
occurs at a temperature of about 50.degree. C. and the acylation
occurs at a temperature of about 50.degree. C.
[0211] An especially preferred aspect of the X method is
wherein
[0212] the alcohol is methanol;
[0213] the acid is HCl;
[0214] the amine is methylamine;
[0215] R.sup.2 is H; and
[0216] R.sup.3 is chloro.
[0217] An especially preferred aspect of the X method is
wherein
[0218] the alcohol is methanol;
[0219] the acid is HCl;
[0220] the amine is methylamine;
[0221] R.sup.2 is cyclopropyl; and
[0222] R.sup.3 is fluoro.
[0223] An especially preferred aspect of the X method is
wherein
[0224] the alcohol is methanol;
[0225] the acid is HCl;
[0226] the amine is methylamine;
[0227] R.sup.2 is H; and
[0228] R.sup.3 is trifluoromethyl.
[0229] Another aspect of this invention are methods of treating a
mammal (e.g., human) having a disease or condition mediated by an
A.sub.3 adenosine receptor by administering a therapeutically
effective amount of a compound of Formula I, a prodrug thereof, or
a pharmaceutically acceptable salt of said compound or of said
prodrug to the mammal.
[0230] Another aspect of this invention is directed to methods of
reducing tissue damage (e.g., substantially preventing tissue
damage, inducing tissue protection) resulting from ischemia or
hypoxia comprising administering to a mammal (e.g., a female or
male human) in need of such treatment a therapeutically effective
amount of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug.
[0231] Preferred ischemic/hypoxic tissues taken individually or as
a group are wherein the ischemic/hypoxic tissue is cardiac, brain,
liver, kidney, lung, gut, skeletal muscle, spleen, pancreas, nerve,
spinal cord, retina tissue, the vasculature, or intestinal
tissue.
[0232] An especially preferred ischemic/hypoxic tissue is cardiac
tissue.
[0233] It is especially preferred that the compounds are
administered to prevent perioperative myocardial ischemic
injury.
[0234] Preferably, the compounds of this invention are administered
prophylactically.
[0235] The ischemic/hypoxic damage may occur during organ
transplantation.
[0236] Preferably, the compounds of this invention are administered
prior to, during or shortly after, cardiac surgery or non-cardiac
surgery (e.g., a three to four day infusion).
[0237] In one aspect of this invention a compound of Formula I is
administered locally.
[0238] A preferred dosage is about 0.001 to 100 mg/kg/day of the
Formula I compound, a prodrug thereof or a pharmaceutically
acceptable salt of said compound or of said prodrug. An especially
preferred dosage is about 0.01 to 50 mg/kg/day of a compound of
Formula I, a prodrug thereof, or a pharmaceutically acceptable salt
of said compound or of said prodrug.
[0239] Another aspect of this invention is directed to methods of
reducing myocardial tissue damage (e.g., substantially preventing
tissue damage, inducing tissue protection) during surgery (e.g.,
coronary artery bypass grafting (CABG) surgeries, vascular
surgeries, percutaneous transluminal coronary angioplasty (PTCA) or
any percutaneous transluminal coronary intervention (PTCI), organ
transplantation, or other non-cardiac surgeries) comprising
administering to a mammal (e.g., a female or male human) a
therapeutically effective amount of a compound of Formula I, a
prodrug thereof, or a pharmaceutically acceptable salt of said
compound or of said prodrug.
[0240] Another aspect of this invention is directed to methods of
reducing myocardial tissue damage (e.g., substantially preventing
tissue damage, inducing tissue protection) in patients presenting
with ongoing cardiac (acute coronary syndromes, e.g., myocardial
infarction or unstable angina) or cerebral ischemic events (e.g.,
stroke) comprising administering to a mammal (e.g., a female or
male human) a therapeutically effective amount of a compound of
Formula I, a prodrug thereof, or a pharmaceutically acceptable salt
of said compound or of said prodrug.
[0241] Another aspect of this invention is directed to chronic
methods of reducing myocardial tissue damage (e.g., substantially
preventing tissue damage, inducing tissue protection) in a patient
with diagnosed coronary heart disease (e.g., previous myocardial
infarction or unstable angina) or patients who are at high risk for
myocardial infarction (e.g., age>65 and two or more risk factors
for coronary heart disease) comprising administering to a mammal
(e.g., a female or male human) a therapeutically effective amount
of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug.
[0242] Another aspect of this invention is directed to methods of
preventing ischemic/hypoxic damage comprising the chronic oral
administration to a mammal in need of such treatment of a
therapeutically effective amount of a compound of Formula I, a
prodrug of said compound, or a pharmaceutically acceptable salt of
said compound or of said prodrug.
[0243] Another aspect of this invention is directed to methods for
treating cardiovascular diseases comprising administering to a
mammal (e.g., a female or male human) a therapeutically effective
amount of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug.
[0244] Another aspect of this invention is directed to methods for
treating arteriosclerosis comprising administering to a mammal
(e.g., a female or male human) a therapeutically effective amount
of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug.
[0245] Another aspect of this invention is directed to methods for
treating arrhythmia comprising administering to a mammal (e.g., a
female or male human) a therapeutically effective amount of a
compound of Formula I, a prodrug thereof, or a pharmaceutically
acceptable salt of said compound or of said prodrug.
[0246] Another aspect of this invention is directed to methods for
treating angina pectoris comprising administering to a mammal
(e.g., a female or male human) a therapeutically effective amount
of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug.
[0247] Another aspect of this invention is directed to methods for
treating cardiac hypertrophy comprising administering to a mammal
(e.g., a female or male human) a therapeutically effective amount
of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug.
[0248] Another aspect of this invention is directed to methods for
treating renal diseases comprising administering to a mammal (e.g.,
a female or male human) a therapeutically effective amount of a
compound of Formula I, a prodrug thereof, or a pharmaceutically
acceptable salt of said compound or of said prodrug.
[0249] Another aspect of this invention is directed to methods for
treating diabetic complications comprising administering to a
mammal (e.g., a female or male human) a therapeutically effective
amount of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug.
[0250] Another aspect of this invention is directed to methods for
treating restenosis comprising administering to a mammal (e.g., a
female or male human) a therapeutically effective amount of a
compound of Formula I, a prodrug thereof, or a pharmaceutically
acceptable salt of said compound or of said prodrug.
[0251] Another aspect of this invention is directed to methods for
treating organ hypertrophies or hyperplasias comprising
administering to a mammal (e.g., a female or male human) a
therapeutically effective amount of a compound of Formula I, a
prodrug thereof, or a pharmaceutically acceptable salt of said
compound or of said prodrug.
[0252] Another aspect of this invention is directed to methods for
treating septic shock and other inflammatory diseases (septicemia,
endotoxcemia) comprising administering to a mammal (e.g., a female
or male human) a therapeutically effective amount of a compound of
Formula I, a prodrug thereof, or a pharmaceutically acceptable salt
of said compound or of said prodrug.
[0253] Another aspect of this invention is directed to methods for
treating cerebro ischemic disorders comprising administering to a
mammal (e.g., a female or male human) a therapeutically effective
amount of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug.
[0254] Another aspect of this invention is directed to methods for
treating myocardial stunning comprising administering to a mammal
(e.g., a female or male human) a therapeutically effective amount
of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug.
[0255] Another aspect of this invention is directed to methods for
treating myocardial dysfunction comprising administering to a
mammal (e.g., a female or male human) a therapeutically effective
amount of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug.
[0256] Another aspect of this invention is directed to methods for
treating cerebrovascular diseases comprising administering to a
mammal (e.g., a female or male human) a therapeutically effective
amount of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug.
[0257] Another aspect of this invention is directed to methods for
treating organ hypertrophies or hyperplasias comprising
administering to a mammal (e.g., a female or male human) a
therapeutically effective amount of a compound of Formula I, a
prodrug thereof, or a pharmaceutically acceptable salt of said
compound or of said prodrug.
[0258] This invention is also directed to pharmaceutical
compositions which comprise a therapeutically effective amount of a
compound of Formula I, a prodrug thereof, or a pharmaceutically
acceptable salt of said compound or of said prodrug and a
pharmaceutically acceptable carrier, vehicle or diluent.
[0259] This invention is also directed to pharmaceutical
compositions for the reduction of tissue damage resulting from
ischemia or hypoxia which comprise a therapeutically effective
amount of a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug and a pharmaceutically acceptable carrier, vehicle or
diluent.
[0260] This invention is also directed to a kit for use by a
consumer having or at risk of having a disease or condition
resulting from, for example, ischemia or hypoxia which may be
ameliorated by an A.sub.3 agonist. The kit comprises a) a suitable
dosage form such as an injectable parenteral solution particularly
adapted for intravenous or intramuscular injection comprising a
compound of Formula I; and b) instructions describing a method of
using the dosage form to reduce tissue damage resulting from
ischemia or hypoxia.
[0261] In the above pharmaceutical compositions and methods,
preferred Formula I compounds include the preferred groups of
compounds described above labeled as Group A- to Group Q.
[0262] Yet another aspect of this invention are combinations of a
compound of Formula I, a prodrug thereof, or a pharmaceutically
acceptable salt of said compound or of said prodrug and other
compounds as described below.
[0263] This invention is also directed to a pharmaceutical
combination composition comprising: a therapeutically effective
amount of a composition comprising
[0264] a first compound, said first compound being a compound
Formula I of a prodrug thereof, or a pharmaceutically acceptable
salt of said compound or of said prodrug;
[0265] a second compound, said second compound being a
cardiovascular agent; and, optionally,
[0266] a pharmaceutical carrier, vehicle or diluent.
[0267] Another aspect of this invention are methods of reducing
tissue damage (e.g., substantially preventing tissue damage,
inducing tissue protection) resulting from or which could result
from ischemia or hypoxia comprising administering to a mammal
(e.g., a female or male human)
[0268] a. a first compound, said first compound being a compound of
Formula I, a prodrug thereof, or a pharmaceutically acceptable salt
of said compound or of said prodrug; and
[0269] b. a second compound, said second compound being a
cardiovascular agent
[0270] wherein the amounts of the first and second compounds result
in a therapeutic effect.
[0271] Another aspect of this invention are kits comprising:
[0272] a. a compound of Formula I, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug and a pharmaceutically acceptable carrier, vehicle or
diluent in a first unit dosage form;
[0273] b. a cardiovascular agent and a pharmaceutically acceptable
carrier, vehicle or diluent in a second unit dosage form; and
[0274] c. means for containing said first and second dosage forms
wherein the amounts of the first and second compounds result in a
therapeutic effect.
[0275] In the above combination compositions, combination methods
and kits, preferably the cardiovascular agents and salts thereof
(e.g., agents having a cardiovascular effect) are, for example,
.beta.-blockers (e.g., acebutolol, atenolol, bopindolol, labetolol,
mepindolol, nadolol, oxprenol, pindolol, propranolol, sotalol),
calcium channel blockers (e.g., amlodipine, nifedipine,
nisoldipine, nitrendipine, verapamil), potassium channel openers,
adenosine, adenosine agonists, sodium-hydrogen exchanger type 1
(NHE-1) inhibitors, ACE inhibitors (e.g., captopril, enalapril),
nitrates (e.g., isosorbide dinitrate, isosorbide 5-mononitrate,
glyceryl trinitrate), diuretics (e.g., hydrochlorothiazide,
indapamide, piretanide, xipamide), glycosides (e.g., digoxin,
metildigoxin), thrombolytics (e.g. tPA), platelet inhibitors (e.g.,
reopro), aspirin, dipyridamol, potassium chloride, clonidine,
prazosin, pyruvate dehydrogenase kinase inhibitors (e.g.,
dichloroacetate), pyruvate dehydrogenase complex activators,
biguanides (e.g., metformin) or other adenosine A.sub.3 receptor
agonists. Other cardiovascular agents include angiotensin II (All)
receptor antagonists, C5a inhibitors, soluble complement receptor
type 1 (sCR1) or analogues, partial fatty acid oxidation (PFOX)
inhibitors (specifically, ranolazine), acetyl CoA carboxylase
activators, malonyl CoA decarboxylase inhibitors, 5'AMP-activated
protein kinase (AMPK) inhibitors, adenosine nucleoside inhibitors,
anti-apoptotic agents (e.g., caspase inhibitors), monophosphoryl
lipid A or analogues, nitric oxide synthase activators/inhibitors,
protein kinase C activators (specifically, protein kinase E),
protein kinase delta inhibitor, poly (ADP ribose) synthetase (PARS,
PARP) inhibitors, metformin (gluconeogenesis inhibitors, insulin
sensitizers), endothelin converting enzyme (ECE) inhibitors,
endothelin ETA receptor antagonists, (thrombin activated
fibrinolytic inhibitor) TAFI inhibitors and Na/Ca exchanger
modulators.
[0276] Especially preferred NHE-1 inhibitors are
[1-(8-bromoquinolin-5-yl)-
-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0277]
[1-(6-chloroquinolin-5-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]gua-
nidine;
[0278]
[1-(indazol-7-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0279]
[1-(benzimidazol-5-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidi-
ne;
[0280]
[1-(1-isoquinolyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0281]
[5-cyclopropyl-1-(4-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[0282]
[5-cyclopropyl-1-(quinolin-5-yl)-1H-pyrazole-4-carbonyl]guanidine;
[0283]
[5-cyclopropyl-1-(quinolin-8-yl)-1H-pyrazole-4-carbonyl]guanidine;
[0284]
[1-(indazol-6-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0285]
[1-(indazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0286]
[1-(benzimidazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0287]
[1-(1-methylbenzimidazol-6-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guani-
dine;
[0288]
1-(5-quinolinyl)-5-n-propyl-1H-pyrazole-4-carbonyl]guanidine;
[0289]
[1-(5-quinolinyl)-5-isopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0290]
[5-ethyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[0291]
[1-(2-methylbenzimidazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guani-
dine;
[0292]
[1-(1,4-benzodioxan-6-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0293]
[1-(benzotriazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0294]
[1-(3-chloroindazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0295]
[1-(5-quinolinyl)-5-butyl-1H-pyrazole-4-carbonyl]guanidine;
[0296]
[5-propyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[0297]
[5-isopropyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[0298]
[1-(2-chloro-4-methylsulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-ca-
rbonyl]guanidine;
[0299]
[1-(2-chlorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0300]
[1-(2-trifluoromethyl-4-fluorophenyl)-5-cyclopropyl-1H-pyrazole-4-c-
arbonyl]guanidine;
[0301]
[1-(2-bromophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0302]
[1-(2-fluorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0303]
[1-(2-chloro-5-methoxyphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]-
guanidine;
[0304]
[1-(2-chloro-4-methylaminosulfonylphenyl)-5-cyclopropyl-1H-pyrazole-
-4-carbonyl]guanidine;
[0305]
[1-(2,5-dichlorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanid-
ine;
[0306]
[1-(2,3-dichlorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanid-
ine;
[0307]
[1-(2-chloro-5-aminocarbonylphenyl)-5-cyclopropyl-1H-pyrazole-4-car-
bonyl]guanidine;
[0308]
[1-(2-chloro-5-aminosulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-car-
bonyl]guanidine;
[0309]
[1-(2-fluoro-6-trifluoromethylphenyl)-5-cyclopropyl-1H-pyrazole-4-c-
arbonyl]guanidine;
[0310]
[1-(2-chloro-5-methylsulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-ca-
rbonyl]guanidine;
[0311]
[1-(2-chloro-5-dimethylaminosulfonylphenyl)-5-cyclopropyl-1H-pyrazo-
le-4-carbonyl]guanidine;
[0312]
[1-(2-trifluoromethyl-4-chlorophenyl)-5-cyclopropyl-1H-pyrazole-4-c-
arbonyl]guanidine;
[0313]
[1-(2-chlorophenyl)-5-methyl-1H-pyrazole-4-carbonyl]guanidine;
[0314]
[5-methyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carbonyl]guanid-
ine;
[0315] [5-ethyl-1-phenyl-1H-pyrazole-4-carbonyl]guanidine;
[0316]
[5-cyclopropyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carbonyl]g-
uanidine;
[0317]
[5-cyclopropyl-1-phenyl-1H-pyrazole-4-carbonyl]guanidine;
[0318]
[5-cyclopropyl-1-(2,6-dichlorophenyl)-1H-pyrazole-4-carbonyl]guanid-
ine; and pharmaceutically acceptable salts thereof.
[0319] In the above combination compositions, combination methods
and kits preferred Formula I compounds include the preferred groups
of compounds described above labeled as Group A to Group Q.
[0320] This invention is also directed to a pharmaceutical
combination composition comprising: a therapeutically effective
amount of a composition comprising
[0321] a first compound, said first compound being a Formula I
compound, a prodrug thereof, or a pharmaceutically acceptable salt
of said compound or of said prodrug;
[0322] a second compound, said second compound being a glycogen
phosphorylase inhibitor; and, optionally,
[0323] a pharmaceutical carrier, vehicle or diluent.
[0324] Another aspect of this invention are methods of reducing
tissue damage (e.g., substantially preventing tissue damage,
inducing tissue protection) resulting from or which could result
from ischemia or hypoxia comprising administering to a mammal
(e.g., a female or male human)
[0325] a. a first compound, said first compound being a Formula I
compound, a prodrug thereof, or a pharmaceutically acceptable salt
of said compound or of said prodrug; and
[0326] b. a second compound, said second compound being a glycogen
phosphorylase inhibitor
[0327] wherein the amounts of the first and second compounds result
in a therapeutic effect.
[0328] Another aspect of this invention are kits comprising:
[0329] a. a Formula I compound, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug and a pharmaceutically acceptable carrier, vehicle or
diluent in a first unit dosage form;
[0330] b. a glycogen phosphorylase inhibitor and a pharmaceutically
acceptable carrier, vehicle or diluent in a second unit dosage
form; and
[0331] c. means for containing said first and second dosage forms
wherein the amounts of the first and second compounds result in a
therapeutic effect.
[0332] In the above combination compositions, combination methods
and kits preferred Formula I compounds include the preferred groups
of compounds described above labeled as Group A to Group Q.
[0333] In the above combination compositions, combination methods
and kits preferred glycogen phosphorylase inhibitors are
[0334] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-((R)-hydroxy-dimethylcar-
bamoyl-methyl)-2-phenyl-ethyl]-amide,
[0335] 5,6-dichloro-1H-indole-2-carboxylic acid
{(1S)-[(R)hydroxy-(methoxy-
-methyl-carbamoyl)-methyl]-2-phenyl-ethyl}-amide,
[0336] 5-chloro-1H-indole-2-carboxylic acid
{(1S)-[(R)-hydroxy-(methoxy-me-
thyl-carbamoyl)-methyl]-2-phenyl-ethyl}-amide,
[0337] 5-chloro-1H-indole-2-carboxylic acid
((1S)-{(R)-hydroxy-[(2-hydroxy-
-ethyl)-methyl-carbamoyl]-methyl}-2-phenyl-ethyl)amide,
[0338] 5-chloro-1H-indole-2-carboxylic acid
{(1S)-[(R)-hydroxy-(methyl-pyr-
idin-2-yl-carbamoyl)-methyl]-2-phenyl-ethyl}-amide,
[0339] 5-chloro-1H-indole-2-carboxylic acid
((1S)-{(R)-hydroxy-[methyl-(2--
pyridin-2-yl-ethyl)-carbamoyl]-methyl}-2-phenyl-ethyl)-amide,
[0340] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-(2R)-hydroxy-3-(4-
-methyl-piperazin-1-yl)-3-oxo-propyl]-amide hydrochloride,
[0341] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-(2R)-hydroxy-3-(3-
-hydroxy-azetidin-1-yl)-3-oxo-propyl]-amide,
[0342] 5-chloro-1H-indole-2-carboxylic acid
((1S)-benzyl-(2R)-hydroxy-3-is-
oxazolidin-2-yl-3-oxo-propyl)-amide,
[0343] 5-chloro-1H-indole-2-carboxylic acid
((1S)-benzyl-(2R)-hydroxy-3-[1-
,2]oxazinan-2-yl-3-oxo-propyl)-amide,
[0344] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-(2R)-hydroxy-3-((-
3S)-hydroxy-pyrrolidin-1-yl)-3-oxo-propyl]-amide,
[0345] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-3-((3S,4S)-dihydr-
oxy-pyrrolidin-1-yl)-(2R)-hydroxy-3-oxo-propyl]-amide,
[0346] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-3-((3R,4S)-dihydr-
oxy-pyrrolidin-1-yl)-(2R)-hydroxy-3-oxo-propyl]-amide,
[0347] 5-chloro-1H-indole-2-carboxylic acid
((1S)-benzyl-(2R)-hydroxy-3-mo-
rpholin-4-yl-3-oxo-propyl)-amide,
[0348] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-(3-hydroxyimino-
-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,
[0349] 5-chloro-1H-indole-2-carboxylic acid
[2-(cis-3,4-dihydroxy-pyrrolid- in-1-yl)-2-oxo-ethyl]-amide,
[0350] 5-chloro-1H-indole-2-carboxylic acid
[2-((3S,4S)-dihydroxy-pyrrolid- in-1-yl)-2-oxo-ethyl]-amide,
[0351] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-(cis-3,4-dihydr-
oxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,
[0352] 5-chloro-1H-indole-2-carboxylic acid
[2-(1,1-dioxo-thiazolidin-3-yl- )-2-oxo-ethyl]-amide,
[0353] 5-chloro-1H-indole-2-carboxylic acid
(2-oxo-2-thiazolidin-3-yl-ethy- l)-amide,
[0354] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-(4-fluoro-benzyl)-2-(4-h-
ydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide,
[0355] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-((3RS)-hydroxy--
piperidin-1-yl)-2-oxo-ethyl]-amide,
[0356] 5-chloro-1H-indole-2-carboxylic acid
[2-oxo-2-((1RS)-oxo-1-thiazoli- din-3-yl)-ethyl]-amide,
[0357] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-(2-fluoro-benzyl)-2-(4-h-
ydroxy-piperidin-1-yl)-2-oxo-ethyl]-amide,
[0358] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-((3S,4S)-dihydr-
oxy-pyrrolidin-1-yl)-2-oxo-ethyl]-amide,
[0359] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-(3-hydroxy-azet- idin-1-yl)-2-oxo-ethyl]-amide,
[0360] 5-Chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-(3-hydroxyimino- -azetidin-1-yl)-2-oxo-ethyl]-amide
or
[0361] 5-chloro-1H-indole-2-carboxylic acid
[(1S)-benzyl-2-(4-hydroxyimino-
-piperidin-1-yl)-2-oxo-ethyl]-amide.
[0362] This invention is also directed to a pharmaceutical
combination composition comprising: a therapeutically effective
amount of a composition comprising
[0363] a first compound, said first compound being a Formula I
compound, a prodrug thereof, or a pharmaceutically acceptable salt
of said compound or of said prodrug;
[0364] a second compound, said second compound being an aldose
reductase inhibitor; and, optionally,
[0365] a pharmaceutical carrier, vehicle or diluent.
[0366] Another aspect of this invention are methods of reducing
tissue damage (e.g., substantially preventing tissue damage,
inducing tissue protection) resulting from or which could result
from ischemia or hypoxia comprising administering to a mammal
(e.g., a female or male human)
[0367] a. a first compound, said first compound being a Formula I
compound, a prodrug thereof, or a pharmaceutically acceptable salt
of said compound or of said prodrug; and
[0368] b. a second compound, said second compound being an aldose
reductase inhibitor
[0369] wherein the amounts of the first and second compounds result
in a therapeutic effect.
[0370] Another aspect of this invention are kits comprising:
[0371] a. a Formula I compound, a prodrug thereof, or a
pharmaceutically acceptable salt of said compound or of said
prodrug and a pharmaceutically acceptable carrier, vehicle or
diluent in a first unit dosage form;
[0372] b. an aldose reductase inhibitor and a pharmaceutically
acceptable carrier, vehicle or diluent in a second unit dosage
form; and
[0373] c. means for containing said first and second dosage forms
wherein the amounts of the first and second compounds result in a
therapeutic effect.
[0374] In the above combination compositions, combination methods
and kits preferred Formula I compounds include the preferred groups
of compounds described above labeled as Group A to Group Q.
[0375] In the above combination compositions, combination methods
and kits a preferred aldose reductase inhibitor is zopolrestat:
1-phthalazineacetic acid,
3,4-dihydro4-oxo-3-[[5-trifluoromethyl)-2-benzo-
thiazolyl]methyl]-.
[0376] In the methods of treatment as applied to the combinations
described above the following are preferred administration routes,
modes, etc.
[0377] Preferred ischemic or hypoxic tissues taken individually or
as a group are wherein the ischemic/hypoxic tissue is cardiac,
brain, liver, kidney, lung, gut, skeletal muscle, spleen, pancreas,
nerve, spinal cord, retina tissue, the vasculature, or intestinal
tissue.
[0378] An especially preferred ischemic or hypoxic tissue is
cardiac tissue.
[0379] It is especially preferred that the combinations are
administered to prevent perioperative myocardial ischemic
injury.
[0380] Preferably, the combinations of this invention are
administered prophylactically.
[0381] The ischemic/hypoxic damage may occur during organ
transplantation.
[0382] Preferably, the combinations of this invention are
administered prior to, during and/or shortly after, cardiac surgery
or non-cardiac surgery.
[0383] In one aspect of this invention the combinations are
administered locally.
[0384] In one aspect of this inventor myocardial tissue damage is
reduced during or after surgery.
[0385] In another aspect of this inventor myocardial tissue damage
is reduced in patients presenting with ongoing cardiac or cerebral
ischemic events.
[0386] In yet another aspect of this inventor myocardial tissue
damage is reduced by chronic administration of the above
combinations in a patient with diagnosed coronary heart
disease.
[0387] The term "reduction" is intended to include partial
prevention or prevention which, although greater than that which
would result from taking no compound or from taking a placebo, is
less than 100% in addition to substantially total prevention.
[0388] The term "damage resulting from ischemia or hypoxia" as
employed herein refers to conditions directly associated with
reduced blood flow or oxygen delivery to tissue, for example due to
a clot or obstruction of blood vessels which supply blood to the
subject tissue and which result, inter alia, in lowered oxygen
transport to such tissue, impaired tissue performance, tissue
dysfunction and/or necrosis and/or apoptosis. Alternatively, where
blood flow or organ perfusion may be quantitatively adequate, the
oxygen carrying capacity of the blood or organ perfusion medium may
be reduced, e.g., in hypoxic environment, such that oxygen supply
to the tissue is lowered, and impaired tissue performance, tissue
dysfunction, and/or tissue necrosis and/or apoptosis ensues.
[0389] The term "treating", "treat" or "treatment" as used herein
includes preventative (e.g., prophylactic) and palliative
treatment.
[0390] By "pharmaceutically acceptable" it is meant the carrier,
diluent, excipients, and/or salt must be compatible with the other
ingredients of the formulation, and not deleterious to the
recipient thereof.
[0391] The expression "prodrug" refers to compounds that are drug
precursors which, following administration, release the drug in
vivo via some chemical or physiological process (e.g., a prodrug on
being brought to the physiological pH or through enzyme action is
converted to the desired drug form).
[0392] Exemplary five to six membered aromatic rings optionally
having one or two heteroatoms selected independently from oxygen,
nitrogen and sulfur are phenyl, furyl, thienyl, pyrrolyl, oxazolyl,
thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl,
pyridinyl, pyridiazinyl, pyrimidinyl and pyrazinyl.
[0393] Exemplary partially saturated, fully saturated or fully
unsaturated five to eight membered rings optionally having one to
three heteroatoms selected independently from oxygen, sulfur and
nitrogen are cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and
phenyl. Further exemplary five membered rings are furyl, thienyl,
pyrrolyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrrolidinyl, 1,3-dioxolanyl,
oxazolyl, thiazolyl, imidazolyl, 2H-imidazolyl, 2-imidazolinyl,
imidazolidinyl, pyrazolyl, 2-pyrazolinyl, pyrazolidinyl,
isoxazolyl, isothiazolyl, 1,2-dithiolyl, 1,3-dithiolyl,
3H-1,2-oxathiolyl, 1,2,3-oxadizaolyl, 1,2,4-oxadiazolyl,
1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl,
1,2,4-trizaolyl, 1,3,4-thiadiazolyl, 3H-1,2,3-dioxazolyl,
1,2,4-dioxazolyl, 1,3,2-dioxazolyl, 1,3,4-dioxazolyl,
5H-1,2,5-oxathiazolyl and 1,3-oxathiolyl.
[0394] Further exemplary six membered rings are 2H-pyranyl,
4H-pyranyl, pyridinyl, piperidinyl, 1,2-dioxinyl, 1,3-dioxinyl,
1,4-dioxanyl, morpholinyl, 1,4-dithianyl, thiomorpholinyl,
pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl, 1,3,5-triazinyl,
1,2,4-triazinyl, 1,2,3-triazinyl, 1,3,5-trithianyl,
4H-1,2-oxazinyl, 2H-1,3-oxazinyl, 6H-1,3-oxazinyl, 6H-1,2-oxazinyl,
1,4-oxazinyl, 2H-1,2-oxazinyl, 4H-1,4-oxazinyl, 1,2,5-oxathiazinyl,
1,4-oxazinyl, o-isoxazinyl, p-isoxazinyl, 1,2,5-oxathiazinyl,
1,2,6-oxathiazinyl and 1,4,2-oxadiazinyl.
[0395] Further exemplary seven membered rings are azepinyl,
oxepinyl, thiepinyl and 1,2,4-diazepinyl.
[0396] Further exemplary eight membered rings are cyclooctyl,
cyclooctenyl and cyclooctadienyl.
[0397] Exemplary bicyclic rings consisting of two fused partially
saturated, fully saturated or fully unsaturated five and/or six
membered rings, taken independently, optionally having one to four
heteroatoms selected independently from nitrogen, sulfur and oxygen
are indolizinyl, indolyl, isoindolyl, indolinyl,
cyclopenta(b)pyridinyl, pyrano(3,4-b)pyrrolyl, benzofuryl,
isobenzofuryl, benzo(b)thienyl, benzo(c)thienyl, 1H-indazolyl,
indoxazinyl, benzoxazolyl, anthranilyl, benzimidazolyl,
benzthiazolyl, purinyl, quinolinyl, isoquinolinyl, cinnolinyl,
phthalazinyl, quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl,
pteridinyl, indenyl, isoindenyl, naphthyl, tetralinyl, decalinyl,
2H-1-benzopyranyl, pyrido(3,4-b)-pyridinyl,
pyrido(3,2-b)-pyridinyl, pyrido(4,3-b)-pyridinyl,
2H-1,3-benzoxazinyl, 2H-1,4-benzoxazinyl, 1H-2,3-benzoxazinyl,
4H-3,1-benzoxazinyl, 2H-1,2-benzoxazinyl and
4H-1,4-benzoxazinyl.
[0398] By alkylene is meant saturated hydrocarbon (straight chain
or branched) wherein a hydrogen atom is removed from each of the
terminal carbons. Exemplary of such groups (assuming the designated
length encompases the particular example) are methylene, ethylene,
propylene, butylene, pentylene, hexylene, heptylene). Of course,
such linking moieties may also be referred to as the substituent
without the "ene" suffix (e.g., methyl) as is commonly done by
those skilled in the art, and still refer to a linking group.
[0399] By halo is meant chloro, bromo, iodo, or fluoro.
[0400] By alkyl is meant straight chain saturated hydrocarbon or
branched saturated hydrocarbon. Exemplary of such alkyl groups
(assuming the designated length encompasses the particular example)
are methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tertiary
butyl, pentyl, isopentyl, neopentyl, tertiary pentyl,
1-methylbutyl, 2-methylbutyl, 3-methylbutyl, hexyl, isohexyl,
heptyl and octyl.
[0401] By alkoxy is meant straight chain saturated alkyl or
branched saturated alkyl bonded through an oxygen. Exemplary of
such alkoxy groups (assuming the designated length encompasses the
particular example) are methoxy, ethoxy, propoxy, isopropoxy,
butoxy, isobutoxy, tertiary butoxy, pentoxy, isopentoxy,
neopentoxy, tertiary pentoxy, hexoxy, isohexoxy, heptoxy and octoxy
.
[0402] As used herein the term mono-N- or
di-N,N-(C.sub.1-C.sub.x)alkyl . . . refers to the
(C.sub.1-C.sub.x)alkyl moiety taken independently when it is
di-N,N-(C.sub.1-C.sub.x)alkyl . . . (x refers to integers).
[0403] It is to be understood that if a carbocyclic or heterocyclic
moiety may be bonded or otherwise attached to a designated
substrate through differing ring atoms without denoting a specific
point of attachment, then all possible points are intended, whether
through a carbon atom or, for example, a trivalent nitrogen atom.
For example, the term "pyridyl" means 2-, 3-, or 4-pyridyl, the
term "thienyl" means 2-, or 3-thienyl, and so forth.
[0404] The expression "pharmaceutically-acceptable salt" refers to
nontoxic anionic salts containing anions such as (but not limited
to) chloride, bromide, iodide, sulfate, bisulfate, phosphate,
acetate, maleate, fumarate, oxalate, lactate, tartrate, citrate,
gluconate, methanesulfonate and 4-toluene-sulfonate. Where more
than one basic moiety exists the expression includes multiple salts
(e.g., di-salt). The expression also refers to nontoxic cationic
salts such as (but not limited to) sodium, potassium, calcium,
magnesium, ammonium or protonated benzathine
(N,N'-dibenzylethylenediamine), choline, ethanolamine,
diethanolamine, ethylenediamine, meglamine (N-methyl-glucamine),
benethamine (N-benzylphenethylamine), piperazine or tromethamine
(2-amino-2-hydroxymethyl-1,3-propanediol).
[0405] As used herein, the expressions "reaction-inert solvent" and
"inert solvent" refers to a solvent or mixture of solvents which
does not interact with starting materials, reagents, intermediates
or products in a manner which adversely affects the yield of the
desired product.
[0406] The chemist of ordinary skill will recognize that certain
compounds of this invention will contain one or more atoms which
may be in a particular stereochemical or geometric configuration,
giving rise to stereoisomers and configurational isomers. All such
isomers and mixtures thereof are included in this invention.
Hydrates of the compounds of this invention are also included.
[0407] DMF means N,N-dimethylformamide. DMSO means dimethyl
sulfoxide. THF means tetrahydrofuran.
[0408] The subject invention also includes isotopically-labelled
compounds, which are identical to those recited in Formula I, but
for the fact that one or more atoms are replaced by an atom having
an atomic mass or mass number different from the atomic mass or
mass number usually found in nature. Examples of isotopes that can
be incorporated into compounds of the invention include isotopes of
hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and
chlorine, such as .sup.2H, .sup.3H, .sup.13C, .sup.14C, .sup.15N,
.sup.18O, .sup.17O, .sup.31P, .sup.32P, .sup.35S, .sup.18F, and
.sup.36Cl, respectively. Compounds of the present invention,
prodrugs thereof, and pharmaceutically acceptable salts of said
compounds or of said prodrugs which contain the aforementioned
isotopes and/or other isotopes of other atoms are within the scope
of this invention. Certain isotopically-labelled compounds of the
present invention, for example those into which radioactive
isotopes such as .sup.3H and .sup.14C are incorporated, are useful
in drug and/or substrate tissue distribution assays. Tritiated,
i.e., .sup.3H, and carbon-14, i.e., .sup.14C, isotopes are
particularly preferred for their ease of preparation and
detectability. Further, substitution with heavier isotopes such as
deuterium, i.e., .sup.2H, can afford certain therapeutic advantages
resulting from greater metabolic stability, for example increased
in vivo half-life or reduced dosage requirements and, hence, may be
preferred in some circumstances. Isotopically labelled compounds of
Formula I of this invention and prodrugs thereof can generally be
prepared by carrying out the procedures disclosed in the Schemes
and/or in the Examples below, by substituting a readily available
isotopically labelled reagent for a non-isotopically labelled
reagent.
[0409] Other features and advantages will be apparent from this
description and claims which describe the invention.
DETAILED DESCRIPTION OF THE INVENTION
[0410] In general the compounds of this invention can be made by
processes which include processes analogous to those known in the
chemical arts, particularly in light of the description contained
herein. Certain processes for the manufacture of the compounds of
this invention are provided as further features of the invention
and are illustrated by the following reaction schemes. Other
processes are described in the experimental section. 11 12 13
14
[0411] In general, the compounds of this invention can be made by
coupling the desired chloropurine riboside and benzyl amine
followed by azide reduction. The following text which is keyed to
the above SCHEMES, provides a more detailed description.
[0412] According to reaction SCHEME I, the desired Formula I
compounds wherein R.sup.1, R.sup.2, R.sup.3, X, Y, Z, D and G are
as defined above may be prepared by reduction of the azide in the
corresponding Formula II compound. Typically the reduction is
accomplished by combining the Formula II compound with a trialkyl
or triaryl phosphine, preferably triphenyl phosphine, in a reaction
inert solvent such as tetrahydrofuran, at temperatures of about
0.degree. C. to about 65.degree. C., typically at ambient
temperature, for about thirty minutes to about two hours. The
reaction is then treated with a base, preferably an amine base,
most preferably ammonium hydroxide for about six hours to about
forty-eight hours at a temperature of about 0.degree. C. to about
65.degree. C., preferably at ambient temperature.
[0413] The desired Formula II compound wherein R.sup.2, R.sup.3, X,
Y, Z, D and G are as defined above and R.sup.1 is an ester may be
prepared from the appropriate Formula III compound and benzyl amine
derivative (wherein R.sup.3, D and G are as defined above).
Typically, the condensation reaction is run in a polar solvent,
such as ethanol, in the presence of a base, preferably an amine
base, most preferably triethylamine at elevated temperatures of
about 40.degree. C. to about 75.degree. C. for about two hours to
about twenty-four hours.
[0414] Analogously, the desired Formula II compound wherein
R.sup.2, R.sup.3, X, Y, Z, D and G are as defined above and R.sup.1
is an amide may be prepared from the appropriate Formula VI
compound and benzyl amine derivative (wherein R.sup.3, D and G are
as defined above). Typically, the condensation reaction is run in a
polar solvent, such as ethanol, in the presence of a base,
preferably an amine base, most preferably triethylamine at elevated
temperatures of about 40.degree. C. to about 75.degree. C. for
about two hours to about twenty-four hours.
[0415] The Formula VI amide may be prepared from the corresponding
Formula III ester by amine addition. Typically, the appropriate
amine is added to the Formula III ester at a temperature of about
15.degree. C. to about 50.degree. C. for about one hour to about
twenty-four hours in a polar solvent such as methanol.
[0416] Some of the methods useful for the preparation of the
compounds described herein may require protection of remote
functionality (e.g., primary amine, secondary amine, carboxyl in
Formula I precursors). The need for such protection will vary
depending on the nature of the remote functionality and the
conditions of the preparation methods. The need for such protection
is readily determined by one skilled in the art. The use of such
protection/deprotection methods is also within the skill in the
art. For a general description of protecting groups and their use,
see T. W. Greene, Protective Groups in Organic Synthesis, John
Wiley & Sons, New York, 1991
[0417] Thus, for example, in an alternative reaction sequence the
desired Formula I compound wherein R.sup.1, R.sup.2, R.sup.3, X, Y,
Z, D and G are as defined above may be prepared from the
corresponding Formula VI compound by protection and amine addition
followed by deprotection. Thus, the Formula VI compound wherein
R.sup.1, R.sup.2, X, Y and Z are as defined above undergoes azide
reduction. Typically the reduction is accomplished by combining the
Formula VI compound with a trialkyl or triaryl phosphine,
preferably triphenyl phosphine, in a reaction inert solvent such as
tetrahydrofuran, at temperatures of about 0.degree. C. to about
65.degree. C., typically at ambient temperature, for about thirty
minutes to about two hours. The reaction is then treated with a
base, preferably an amine base, most preferably ammonium hydroxide
for about six hours to about forty-eight hours at a temperature of
about 0.degree. C. to about 65.degree. C. Following reduction, the
amine moiety is protected (P.sup.1).
[0418] Preferably the amine is protected with a tert-butoxycarbonyl
group. The protection is accomplished by treating the amine with
tert-butoxycarbonyl anhydride and a base, preferably an amine base,
most preferably triethylamine, in an anhydrous solvent such as
dichloromethane, at ambient temperature for about five hours to
about twenty-four hours.
[0419] The desired Formula IV compound wherein R.sup.1, R.sup.2,
R.sup.3, X, Y, Z, D and G are as defined above are prepared from
the appropriate Formula V compound and benzyl amine derivative
(wherein R.sup.3, D and G are as defined above). Typically, the
condensation reaction is run in a polar solvent, such as ethanol,
in the presence of a base, preferably an amine base, most
preferably triethylamine at elevated temperatures of about
40.degree. C. to about 75.degree. C. for about two hours to about
twenty-four hours.
[0420] Following amine addition the desired Formula I compound may
be prepared from the corresponding protected Formula IV compound by
an appropriate catalyzed deprotection reaction. Typically, the
protected (e.g., tertiary butyl ester protected) compound is
treated with a strong acid, preferably trifluoroacetic acid at
10.degree. C. to 50.degree. C., preferably at ambient temperature,
for about one hour to about eight hours to remove the protecting
moiety.
[0421] According to reaction SCHEME II the desired Formula IA
compounds wherein R.sup.1, R.sup.2, R.sup.3, X, Y, Z, R.sup.4 and
R.sup.5 are as defined above and D is oxy, thio or NH may be
prepared by reduction of the azide in the corresponding Formula XX
compound. Typically, the reduction is accomplished by combining the
Formula XX compound with a trialkyl or triaryl phosphine,
preferably triphenyl phosphine, in a reaction inert solvent such as
tetrahydrofuran, at temperatures of about 0.degree. C. to about
65.degree. C., typically at ambient temperature, for about thirty
minutes to about two hours. The reaction is then treated with a
base, preferably an amine base, most preferably ammonium hydroxide
for about six hours to about forty-eight hours at a temperature of
about 0.degree. C. to about 65.degree. C.
[0422] The Formula XXI ester (following conversion to the acid) is
coupled with the appropriate amine in the presence of a suitable
coupling agent to prepare the desired Formula XX compound. A
suitable coupling agent is one which transforms a carboxylic acid
into a reactive species which forms an amide linkage on reaction
with an amine.
[0423] The coupling agent may be a reagent which effects this
condensation in a one pot process when mixed together with the
carboxylic acid and amine. Exemplary coupling reagents are
1-(3-dimethylaminopropyl)-3-ethylc- arbodiimide
hydrochloride-hydroxybenzotriazole (EDC/HOBT),
dicyclohexylcarbodiimide/hydroxybenzotriazole(HOBT),
2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ), and
diethylphosphorylcyanide. The coupling is performed in an inert
solvent, preferably an aprotic solvent at a temperature of about
-20.degree. C. to about 50.degree. C. for about one to about
forty-eight hours, in the presence of excess amine as base.
Exemplary solvents include acetonitrile, dichloromethane,
dimethylformamide and chloroform or mixtures thereof.
[0424] The coupling agent may also be that agent which converts the
carboxylic acid to an activated intermediate which is isolated
and/or formed in a first step and allowed to react with the amine
in a second step. Examples of such coupling agents and activated
intermediates are thionyl chloride or oxalyl chloride to form the
acid chloride, cyanuric fluoride to form an acid fluoride or an
alkyl chloroformate such as isobutyl or isopropenyl chloroformate
or propanephosphonic anhydride (propanephosphonic acid anhydride,
PPA) (with a tertiary amine base) to form a mixed anhydride of the
carboxylic acid, or carbonyldiimidazole to form an acylimidazole.
If the coupling agent is oxalyl chloride, it is advantageous to
employ a small amount of dimethylformamide as cosolvent with
another solvent (such as dichloromethane) to catalyze the formation
of the acid chloride. This activated acid derivative may be coupled
by mixing with excess amine in an appropriate solvent together with
an appropriate base. Appropriate solvent/base combinations are, for
example, dichloromethane, dimethylformamide or acetonitrile or
mixtures thereof in the presence of excess amine as base. Other
appropriate solvent/base combinations include water or a
(C.sub.1-C.sub.5)alcohol or a mixture thereof together with a
cosolvent such as dichloromethane, tetrahydrofuran or dioxane and a
base such as sodium, potassium or lithium hydroxide in sufficient
quantity to consume the acid liberated in the reaction. Use of
these coupling agents and appropriate selection of solvents and
temperatures are known to those skilled in the art or can be
readily determined from the literature. These and other exemplary
conditions useful for coupling carboxylic acids are described in
Houben-Weyl, Vol XV, part II, E. Wunsch, Ed., G. Theime Verlag,
1974, Stuttgart; M. Bodansky, Principles of Peptide Synthesis,
Springer-Verlag, Berlin 1984; and The Peptides. Analysis, Synthesis
and Biology (ed. E. Gross and J. Meienhofer), vols 1-5 (Academic
Press, NY 1979-1983).
[0425] The Formula XXI compound may be converted to the
corresponding acid by an acid catalyzed deprotection. Typically the
protected (e.g., tertiary butyl ester protected) compound is
treated with a strong acid, preferably trifluoroacetic acid at
10.degree. C. to 50.degree. C., preferably at ambient temperature
for about one hour to about eight hours to form the corresponding
acid.
[0426] The desired Formula XXI compound wherein R.sup.1, R.sup.2,
R.sup.3, X, Y, Z, D and R is a convenient ester moiety (alkyl) may
be prepared from the appropriate Formula XXII compounds wherein
R.sup.1, R.sup.2, R.sup.3, X, Y, Z and D are as described above
(typically DH represents D as oxy, thio or NH, i.e., prior to
alkylation) by an alkylation reaction. Generally, the Formula XXII
compound is combined with an alkylbromoacetate in the presence of a
strong base such as sodium hydride in a polar aprotic solvent such
as DMF at a temperature of about 0.degree. C. to about 30.degree. C
for a period of about one to about twenty-four hours.
[0427] The Formula XXII compound wherein R.sup.1, R.sup.2, R.sup.3,
X, Y. Z, D are as described above may be prepared from the
appropriate Formula III compound and the appropriate benzyl amine
derivative. Typically, the condensation reaction is performed in a
polar solvent, such as ethanol, in the presence of a base,
preferably an amine base, most preferably triethylamine at elevated
temperatures of about 40.degree. C. to about 75.degree. C. for
about two hours to about twenty-four hours.
[0428] According to SCHEME III the Formula XXX compounds wherein X
and Y are as defined above are prepared from a glycosidation
reaction between the appropriate Formula XXXI compound and a
silylated 6-chloro purine. Typically, the reaction is catalyzed by
a Lewis acid, preferably trimethylsilyltriflate, in a reaction
inert solvent, such as dichloroethane or acetonitrile, at
temperatures from about 30.degree. C. to about 75.degree. C.,
typically at 60.degree. C. for about thirty minutes to about six
hours.
[0429] The desired Formula XXXI compounds wherein X is defined
above may be prepared by an acid catalyzed hydrolysis of the
appropriate Formula XXXII. compound. Typically the acid is a strong
mineral acid, preferably sulfuric acid, in a protic solvent mixture
of acetic acid and acetic anhydride at a temperature of about
5.degree. C. to about 40.degree. C. for about two hours to about
twenty-four hours.
[0430] Analogously, the desired Formula XXXVI compounds wherein Y
and X are as described above may be prepared from the appropriate
Formula XXXIII compound using the glycosidation and hydrolysis
reactions described above.
[0431] The desired Formula XXXII compound wherein X is defined
above is prepared from the appropriate Formula XXXIII compound by
activation of the carboxylic acid followed by reaction with an
amine. Typically, the Formula XXXIII compound may be activated by
conversion to an acid chloride by, for example, treatment with
oxalyl chloride in a non-polar aprotic solvent, preferably
dichloromethane with a catalytic amount of dimethyl formamide, at a
temperature of about 0.degree. C. with warming to ambient
temperature for about two hours to about eight hours. The acid
chloride is then treated with excess of the appropriate amine at a
temperature of 0.degree. C. to about 30.degree. C.
[0432] The desired Formula XXXIII compound wherein X is defined
above is prepared by oxidation of the appropriate Formula XXXIV
compound. Generally the oxidant is ruthenium tetroxide, prepared
using a catalytic amount of ruthenium trichloride and a
stoichiometric amount of sodium periodate in a solvent mixture of
chloroform, acetonitrile and water. The reaction is conveniently
performed at ambient temperature for about four hours to about
twenty-four hours.
[0433] The desired Formula XXXIV compound wherein X is defined
above is prepared from the appropriate Formula XXXV compound by
treatment with periodic acid which hydrolyzes the isopropylidene
group and cleaves the glycol to furnish the aldehyde. The reaction
is run in ethereal solvents, typically diethyl ether conveniently
at ambient temperature for about two hours to about twenty-four
hours.
[0434] The desired Formula XXXV compound is prepared from the
corresponding hydroxyl compound by activation of the hydroxyl group
and displacement with azide ion. Typically, activation is achieved
by converting the hydroxyl group to the corresponding triflate
derivative by reaction with triflic anhydride in the presence of an
amine base, preferably pyridine at about -30.degree. C. to about
0.degree. C. for about thirty minutes to about two hours. The
resulting triflate is treated with an alkali metal azide,
preferably sodium azide, in a polar aprotic solvent, preferably
dimethylformamide at about ambient temperature to about 50.degree.
C. for about six hours to about twenty-four hours.
[0435] SCHEME IV provides preparation methods for the benzylamine
intermediates of this invention.
[0436] Thus, the Formula L benzyl amines wherein D, G and R.sup.3
are as described above (typically DH represents D as oxy, thio or
NH, i.e., prior to alkylation) are prepared for example by one of
three methods. In the first method, the Formula LII imide is
prepared from the corresponding Formula LI aromatic compound
through an imidoalkylation reaction. Thus, the appropriate Formula
LI compound is treated with N-chloromethylphthalimide and an acid
or Lewis acid catalyst, such as zinc chloride in an aprotic
reaction inert solvent such as THF at a temperature of ambient
temperature to about 100.degree. C., preferably about 50.degree.
C.
[0437] The resulting Formula LII compound is alkylated to prepare
the corresponding Formula L compound by combination with the
appropriate alkylation agent in a polar aprotic solvent such as DMF
at a temperature of about 0.degree. C. to about 50.degree. C. for a
period of about two to about twenty-four hours. Alternatively the
alkylation can be accomplished under Mitsunobu conditions using an
appropriate alcohol, triphenylphosphine, diethylazadicarboxylate in
an ether solvent, preferably THF at ambient temperature for about
four hours to about twenty-four hours.
[0438] The resulting Formula LIX phthalimide is deprotected by
treatment with hydrazine hydrate in a protic solvent such as
ethanol at a temperature of ambient temperature to about
100.degree. C., preferably about 50.degree. C. for about one to
about six hours. Alternatively, the deprotection can be
accomplished by first reducing the imide with a hydride reducing
agent, preferably sodium borohydride, followed by heating with
acetic acid at about 50.degree. C. to about 100.degree. C. for
about ten to about twenty-four hours.
[0439] Alternatively, the Formula L benzyl amine can be prepared
from a reaction sequence as follows. The Formula LV compounds are
prepared from the corresponding Formula LII compounds through a two
step procedure of alkylation followed by metal hydride reduction.
The Formula LV compounds can also be prepared from the
corresponding Formula LIV compounds by treatment with a boronic
acid, preferably phenylboronic acid, and formaldehye and an acid
catalyst such as propionic acid in an aprotic solvent such as
benzene at a temperature of about 30.degree. C. to about
100.degree. C. for about one to about twenty-four hours.
[0440] The Formula LVI compound wherein D, G and R.sup.3 can be
prepared from the corresponding Formula LV compound. Generally, the
LV benzyl alcohol is treated with diphenylphosphoryl azide and a
base, preferably a strong amine base such as diazobicycloundecane
(DBU), in an aprotic solvent, preferably toluene, at temperatures
from about 0.degree. C. to about 50.degree. C., most preferably at
ambient temperature for about one to about twenty-four hours.
[0441] The resulting azide may be reduced to prepare the Formula L
compound. In general, the reduction is accomplished by treating the
appropriate Formula LVI azide with a hydrogenation catalyst,
preferably a palladium catalyst, most preferably 10% Pd on carbon,
in a reaction inert solvent such as an alcohol solvent, preferably
ethanol. The reaction vessel is placed under an atmosphere of
hydrogen gas, preferably at 15 to 50 psi conveniently at ambient
temperature for about thirty minutes to about four hours. The azide
reduction can also be accomplished by combining the azide with a
trialkyl or triaryl phosphine, preferably triphenyl phosphine, in a
reaction inert solvent such as tetrahydrofuran, at a temperature of
about 0.degree. C. to about 65.degree. C., typically at ambient
temperature, for about one-half hour to about two hours. The
reaction is then treated with a base, preferably an amine base,
most preferably ammonium hydroxide for about six hours to about
forty-eight hours.
[0442] A third method for the synthesis of Formula L compounds is
from the Formula LVII nitrites by either catalytic hydrogenation or
metal hydride reduction. In general, the catalytic hydrogenation is
accomplished by treating the appropriate formula LVII compound with
a hydrogenation catalyst, preferably Raney nickel, in a reaction
inert solvent such as an alcohol solvent, preferably ethanol
containing about 1% aqueous ammonium hydroxide solution. The
reaction vessel is placed under an atmosphere of hydrogen gas,
preferably at 15 to 50 psi conveniently at ambient temperature for
about 30 minutes to about four hours. Alternatively the reduction
can be performed using a metal hydride reducing agent, preferably
lithium aluminum hydride in an ethereal solvent, preferably
tetrahydrofuran, at a temperature of about 0.degree. C. to about
60.degree. C., preferably at ambient temperature for about one to
about six hours. The Formula LVII compounds are prepared by
alkylation of the corresponding Formula LVIIII compounds by methods
known to those skilled in the art, and as described for Formula LIX
compounds.
[0443] The starting materials and reagents for the above described
compounds are readily available or can be easily synthesized by
those skilled in the art using conventional methods of organic
synthesis. For example, many of the compounds used herein are
related to, or are derived from compounds found in nature, in which
there is a large scientific interest and commercial need, and
accordingly many such compounds are commercially available or are
reported in the literature or are easily prepared from other
commonly available substances by methods which are reported in the
literature.
[0444] Some of the compounds of this invention have asymmetric
carbon atoms and can therefore exist as enantiomers or
diastereomers. Diasteromeric mixtures can be separated into their
individual diastereomers on the basis of their physical chemical
differences by methods known per se., for example, by
chromatography and/or fractional crystallization. Enantiomers can
be separated by converting the enantiomeric mixture into a
diasteromeric mixture by reaction with an appropriate optically
active compound (e.g., alcohol), separating the diastereomers and
converting (e.g., hydrolyzing) the individual diastereomers to the
corresponding pure enantiomers. All such isomers, including
diastereomers, enantiomers and mixtures thereof are considered as
part of this invention. Also, some of the compounds of this
invention are atropisomers (e.g., substituted biaryls) and are
considered as part of this invention.
[0445] Those skilled in the art will recognize that the compounds
of Formula I can exist in several tautomeric forms. All such
tautomeric forms are considered as part of this invention. Also,
for example all enol-keto forms of the compounds of Formula I are
included in this invention.
[0446] Some of the compounds of this invention are acidic and they
form a salt with a pharmaceutically acceptable cation. All of the
compounds of this invention are basic and they form a salt with a
pharmaceutically acceptable anion. All such salts, including
di-salts are within the scope of this invention and they can be
prepared by conventional methods. For example, they can be prepared
simply by contacting the acidic and basic entities, in either an
aqueous, non-aqueous or partially aqueous medium. The salts are
recovered either by filtration, by precipitation with a non-solvent
followed by filtration, by evaporation of the solvent, or, in the
case of aqueous solutions, by lyophilization, as appropriate.
[0447] In addition, when the compounds of this invention form
metabolites, hydrates or solvates they are also within the scope of
the invention.
[0448] Other cardiovascular agents (e.g., agents having a
cardiovascular effect) known to those skilled in the art such as
those described above in the Summary may be used in conjunction
with the compounds of this invention.
[0449] In combination therapy treatment, both the compounds of this
invention and the other drug therapies are administered to mammals
(e.g., humans, male or female) by conventional methods.
[0450] Any NHE-1 inhibitor may be used as the second compound
(active agent) of this invention for combination therapies. The
term NHE-1inhibitor refers to compounds which inhibit the
sodium/proton (Na+/H+) exchange transport system and hence are
useful as a therapeutic or prophylactic agent for diseases caused
or aggravated by the acceleration of the sodium/proton (Na+/H+)
exchange transport system. Such inhibition is readily determined by
those skilled in the art according to standard assays such as are
described herein below and in conventional preclinical
cardioprotection assays [see the in vivo assay in Klein, H. et al.,
Circulation 92:912-917 (1995); the isolated heart assay in Scholz,
W. et al., Cardiovascular Research 29:260-268 (1995); the
antiarrhythmic assay in Yasutake M. et al., Am. J. Physiol.,
36:H2430-H2440 (1994); the NMR assay in Kolke et al., J. Thorac.
Cardiovasc. Surg. 112: 765-775 (1996)]. A variety of NHE-1
inhibitors are described and referenced below, however, other NHE-1
inhibitors will be known to those skilled in the art such as are
disclosed in WO99/43663 published Sep. 2, 1999. Accordingly,
examples of NHE-1 inhibitors useful in the compositions and methods
of this invention include:
[1-(8-bromoquinolin-5-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0451]
[1-(6-chloroquinolin-5-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]gua-
nidine;
[0452]
[1-(indazol-7-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0453]
[1-(benzimidazol-5-yl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidi-
ne;
[0454]
[1-(1-isoquinolyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0455]
[5-cyclopropyl-1-(4-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[0456]
[5-cyclopropyl-1-(quinolin-5-yl)-1H-pyrazole-4-carbonyl]guanidine;
[0457]
[5-cyclopropyl-1-(quinolin-8-yl)-1H-pyrazole-4-carbonyl]guanidine;
[0458]
[1-(indazol-6-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0459]
[1-(indazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0460]
[1-(benzimidazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0461]
[1-(1-methylbenzimidazol-6-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guani-
dine;
[0462]
1-(5-quinolinyl)-5-n-propyl-1H-pyrazole-4-carbonyl]guanidine;
[0463]
[1-(5-quinolinyl)-5-isopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0464]
[5-ethyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[0465]
[1-(2-methylbenzimidazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guani-
dine;
[0466]
[1-(1,4-benzodioxan-6-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0467]
[1-(benzotriazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0468]
[1-(3-chloroindazol-5-yl)-5-ethyl-1H-pyrazole-4-carbonyl]guanidine;
[0469]
[1-(5-quinolinyl)-5-butyl-1H-pyrazole-4-carbonyl]guanidine;
[0470]
[5-propyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[0471]
[5-isopropyl-1-(6-quinolinyl)-1H-pyrazole-4-carbonyl]guanidine;
[0472]
[1-(2-chloro-4-methylsulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-ca-
rbonyl]guanidine;
[0473]
[1-(2-chlorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0474]
[1-(2-trifluoromethyl-4-fluorophenyl)-5-cyclopropyl-1H-pyrazole-4-c-
arbonyl]guanidine;
[0475]
[1-(2-bromophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0476]
[1-(2-fluorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanidine;
[0477]
[1-(2-chloro-5-methoxyphenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]-
guanidine;
[0478]
[1-(2-chloro-4-methylaminosulfonylphenyl)-5-cyclopropyl-1H-pyrazole-
-4-carbonyl]guanidine;
[0479]
[1-(2,5-dichlorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanid-
ine;
[0480]
[1-(2,3-dichlorophenyl)-5-cyclopropyl-1H-pyrazole-4-carbonyl]guanid-
ine;
[0481]
[1-(2-chloro-5-aminocarbonylphenyl)-5-cyclopropyl-1H-pyrazole-4-car-
bonyl]guanidine;
[0482]
[1-(2-chloro-5-aminosulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-car-
bonyl]guanidine;
[0483]
[1-(2-fluoro-6-trifluoromethylphenyl)-5-cyclopropyl-1H-pyrazole-4-c-
arbonyl]guanidine;
[0484]
[1-(2-chloro-5-methylsulfonylphenyl)-5-cyclopropyl-1H-pyrazole-4-ca-
rbonyl]guanidine;
[0485]
[1-(2-chloro-5-dimethylaminosulfonylphenyl)-5-cyclopropyl-1H-pyrazo-
le-4-carbonyl]guanidine;
[0486]
[1-(2-trifluoromethyl-4-chlorophenyl)-5-cyclopropyl-1H-pyrazole-4-c-
arbonyl]guanidine;
[0487]
[1-(2-chlorophenyl)-5-methyl-1H-pyrazole-4-carbonyl]guanidine;
[0488]
[5-methyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carbonyl]guanid-
ine;
[0489] [5-ethyl-1-phenyl-1H-pyrazole-4-carbonyl]guanidine;
[0490]
[5-cyclopropyl-1-(2-trifluoromethylphenyl)-1H-pyrazole-4-carbonyl]g-
uanidine;
[0491]
[5-cyclopropyl-1-phenyl-1H-pyrazole-4-carbonyl]guanidine;
[0492]
[5-cyclopropyl-1-(2,6-dichlorophenyl)-1H-pyrazole-4-carbonyl]guanid-
ine; and
[0493] pharmaceutically acceptable salts thereof.
[0494] Any aldose reductase inhibitor may be used as the second
compound (active agent) of this invention for combination
therapies. The term aldose reductase inhibitor refers to compounds
which inhibit the bioconversion of glucose to sorbitol catalyzed by
the enzyme aldose reductase. Such inhibition is readily determined
by those skilled in the art according to standard assays (J.
Malone, Diabetes, 29:861-864, 1980. "Red Cell Sorbitol, an
Indicator of Diabetic Control"). A variety of aldose reductase
inhibitors are described and referenced below, however, other
aldose reductase inhibitors will be known to those skilled in the
art. The disclosures of U.S. patents listed below are hereby
incorporated by reference. Also, common chemical USAN names or
other designation are in parentheses where applicable, together
with reference to appropriate patent literature disclosing the
compound.
[0495] The activity of an aldose reductase inhibitor in a tissue
can be determined by testing the amount of aldose reductase
inhibitor that is required to lower tissue sorbitol (i.e., by
inhibiting the further production of sorbitol consequent to
blocking aldose reductase) or lower tissue fructose (by inhibiting
the production of sorbitol consequent to blocking aldose reductase
and consequently the production of fructose). While not wishing to
be bound by any particular theory or mechanism, it is believed that
an aldose reductase inhibitor, by inhibiting aldose reductase,
prevents or reduces ischemic or hypoxic damage as described
hereinafter.
[0496] Accordingly, examples of aldose reductase inhibitors useful
in the compositions and methods of this invention include:
[0497] 1.
3-(4-bromo-2-fluorobenzyl)-3,4-dihydro4-oxo-1-phthalazineacetic
acid (ponalrestat, U.S. Pat. No. 4,251,528);
[0498] 2.
N[[(5-trifluoromethyl)-6-methoxy-1-naphthalenyl]thioxomethyl}-N--
methylglycine (tolrestat, U.S. Pat. No. 4,600,724);
[0499] 3.
5-[(Z,E)-.beta.-methylcinnamylidene]-4-oxo-2-thioxo-3-thiazolide-
neacetic acid (epalrestat, U.S. Pat. Nos. 4,464,382, 4,791,126,
4,831,045);
[0500] 4.
3-(4-bromo-2-fluorobenzyl)-7-chloro-3,4-dihydro-2,4-dioxo-1(2H)--
quinazolineacetic acid (zenarestat, U.S. Pat. Nos. 4,734,419, and
4,883,800);
[0501] 5. 2R,4R-6,7-dichloro-4-hydroxy-2-methylchroman-4-acetic
acid (U.S. Pat. No. 4,883,410);
[0502] 6.
2R,4R-6,7-dichloro-6-fluoro-4-hydroxy-2-methylchroman-4-acetic acid
(U.S. Pat. No. 4,883,410);
[0503] 7.
3,4-dihydro-2,8-diisopropyl-3-oxo-2H-1,4-benzoxazine-4-acetic acid
(U.S. Pat. No. 4,771,050);
[0504] 8.
3,4-dihydro-3-oxo-4-[(4,5,7-trifluoro-2-benzothiazolyl)methyl]-2-
H-1,4-benzothiazine-2-acetic acid (SPR-210, U.S. Pat. No.
5,252,572);
[0505] 9.
N-[3,5-dimethyl-4-[(nitromethyl)sulfonyl]phenyl]-2-methyl-benzen-
eacetamide (ZD5522, U.S. Pat. Nos. 5,270,342 and 5,430,060);
[0506] 10. (S)-6-fluorospiro[chroman-4,4'-imidazolidine]-2,5'-dione
(sorbinil, U.S. Pat. No. 4,130,714);
[0507] 11.
d-2-methyl-6-fluoro-spiro(chroman-4',4'-imidazolidine)-2',5'-di-
one (U.S. Pat. No. 4,540,704);
[0508] 12. 2-fluoro-spiro(9H-fluorene-9,4'imidazolidine)2',5'-dione
(U.S. Pat. No. 4,438,272);
[0509] 13.
2,7-di-fluoro-spiro(9H-fluorene-9,4'imidazolidine)2',5'-dione (U.S.
Pat. Nos. 4,436,745, 4,438,272);
[0510] 14.
2,7-di-fluoro-5-methoxy-spiro(9H-fluorene-9,4'imidazolidine)2',-
5'-dione (U.S. Pat. Nos. 4,436,745, 4,438,272);
[0511] 15.
7-fluoro-spiro(5H-indenol[1,2-b]pyridine-5,3'-pyrrolidine)2,5'--
dione (U.S. Pat. Nos. 4,436,745, 4,438,272);
[0512] 16.
d-cis-6'-chloro-2',3'-dihydro-2'-methyl-spiro-(imidazolidine-4,-
4'-4'-H-pyrano(2,3-b)pyridine)-2,5-dione (U.S. Pat. No.
4,980,357);
[0513] 17.
spiro[imidazolidine-4,5'(6H)-quinoline]2,5-dione-3'-chloro-7,'8-
'-dihydro-7'-methyl-(5'-cis) (U.S. Pat. No. 5,066,659);
[0514] 18.
(2S,4S)-6-fluoro-2',5'-dioxospiro(chroman-4,4'-imidazolidine)2--
carboxamide (U.S. Pat. No. 5,447,946); and
[0515] 19.
2-[(4-bromo-2-fluorophenyl)methyl]-6-fluorospiro[isoquinoline-4-
(1H),3'-pyrrolidine]-1,2',3,5'(2H)-tetrone (ARI-509, U.S. Pat. No.
5,037,831).
[0516] Other aldose reductase inhibitors include compounds having
formula IB 15
[0517] and pharmaceutically acceptable salts thereof, wherein
[0518] Z is O or S;
[0519] R.sup.1 is hydroxy or a group capable of being removed in
vivo to produce a compound of formula IB wherein R.sup.1 is OH;
and
[0520] X and Y are the same or different and are selected from
hydrogen, trifluoromethyl, fluoro, and chloro.
[0521] A preferred subgroup within the above group of aldose
reductase inhibitors includes numbered compounds 1, 2, 3, 4, 5, 6,
9, 10, and 17, and the following compounds of Formula IB:
[0522] 20.
3,4-dihydro-3-(5-fluorobenzothiazol-2-ylmethyl)-4-oxophthalazin-
-1-yl-acetic acid [R.sup.1=hydroxy; X.dbd.F; Y.dbd.H];
[0523] 21.
3-(5,7-difluorobenzothiazol-2-ylmethyl)-3,4-dihydro4-oxophthala-
zin-1-ylacetic acid [R.sup.1=hydroxy; X.dbd.Y.dbd.F];
[0524] 22.
3-(5-chlorobenzothiazol-2-ylmethyl)-3,4-dihydro-4-oxophthalazin-
-1-ylacetic acid [R.sup.1=hydroxy; X.dbd.Cl; Y.dbd.H];
[0525] 23.
3-(5,7-dichlorobenzothiazol-2-ylmethyl)-3,4-dihydro-4-oxophthal-
azin-1-ylacetic acid [R.sup.1=hydroxy; X.dbd.Y.dbd.Cl];
[0526] 24.
3,4-dihydro-4-oxo-3-(5-trifluoromethylbenzoxazol-2-ylmethyl)pht-
halazin-1-ylacetic acid [R.sup.1=hydroxy; X.dbd.CF.sub.3;
Y.dbd.H];
[0527] 25.
3,4-dihydro-3-(5-fluorobenzoxazol-2-ylmethyl)-4-oxophthalazin-1-
-yl-acetic acid [R.sup.1=hydroxy; X.dbd.F; Y.dbd.H];
[0528] 26.
3-(5,7-difluorobenzoxazol-2-ylmethyl)-3,4-dihydro-4-oxophthalaz-
in-1-ylacetic acid [R.sup.1=hydroxy; X.dbd.Y.dbd.F];
[0529] 27.
3-(5-chlorobenzoxazol-2-ylmethyl)-3,4-dihydro-4-oxophthalazin-1-
-ylacetic cid [R.sup.1=hydroxy; X.dbd.Cl; Y.dbd.H];
[0530] 28.
3-(5,7-dichlorobenzoxazol-2-ylmethyl)-3,4-dihydro-4-oxophthalaz-
in-1-ylacetic acid [R.sup.1=hydroxy; X.dbd.Y.dbd.Cl]; and
[0531] 29. zopolrestat; 1-phthalazineacetic acid,
3,4-dihydro-4-oxo-3-[[5--
(trifluoromethyl)-2-benzothiazolyl]methyl]-[R.sup.1=hydroxy;
X=trifluoromethyl; Y.dbd.H].
[0532] In compounds 20-23, and 29 Z is S. In compounds 24-28, Z is
O.
[0533] Of the above subgroup, compounds 20-29 are more preferred
with 29 especially preferred.
[0534] An especially preferred aldose reductase inhibitor is
1-phthalazineacetic acid,
3,4-dihydro-4-oxo-3-[[5-trifluoromethyl)-2-benz-
othiazolyl]methyl]-.
[0535] The aldose reductase inhibitor compounds of this invention
are readily available or can be easily synthesized by those skilled
in the art using conventional methods of organic synthesis,
particularly in view of the pertinent patent specification
descriptions.
[0536] An amount of the aldose reductase inhibitor of this
invention that is effective for the activities of this invention
may be used. Typically, an effective dosage for the aldose
reductase inhibitors for the combination compositions, methods and
kits of this invention is in the range of about 0.1 mg/kg/day to
100 mg/kg/day in single or divided doses, preferably 0.1 mg/kg/day
to 20 mg/kg/day in single or divided doses.
[0537] Any glycogen phosphorylase inhibitor may be used as the
second compound of this invention. The term glycogen phosphorylase
inhibitor refers to any substance or agent or any combination of
substances and/or agents which reduces, retards, or eliminates the
enzymatic action of glycogen phosphorylase. The currently known
enzymatic action of glycogen phosphorylase is the degradation of
glycogen by catalysis of the reversible reaction of a glycogen
macromolecule and inorganic phosphate to glucose-1-phosphate and a
glycogen macromolecule which is one glucosyl residue shorter than
the original glycogen macromolecule (forward direction of
glycogenolysis). Such actions are readily determined by those
skilled in the art according to standard assays (e.g., as described
hereinafter). A variety of these compounds are included in the
following published international patent applications: PCT
application publication WO 96/39384 and WO96/39385. However, other
glycogen phosphorylase inhibitors useful in the combinations,
methods, and kits of this invention will be known to those skilled
in the art.
[0538] Preferred glycogen phosphorylase inhibitors include
compounds having the Formula IC 16
[0539] and the pharmaceutically acceptable salts and prodrugs
thereof
[0540] wherein
[0541] the dotted line ( - - - ) is an optional bond;
[0542] A is --C(H).dbd., --C((C.sub.1-C.sub.4)alkyl).dbd. or
--C(halo).dbd. when the dotted line ( - - - ) is a bond, or A is
methylene or --CH((C.sub.1-C.sub.4)alkyl)- when the dotted line ( -
- - ) is not a bond;
[0543] R.sub.1, R.sub.10 or R.sub.11 are each independently H,
halo, 4-, 6- or 7-nitro, cyano, (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy, fluoromethyl, difluoromethyl or
trifluoromethyl;
[0544] R.sub.2 is H;
[0545] R.sub.3 is H or (C.sub.1-C.sub.5)alkyl;
[0546] R.sub.4 is H, methyl, ethyl, n-propyl,
hydroxy(C.sub.1-C.sub.3)alky- l,
(C.sub.1-C.sub.3)alkoxy(C.sub.1-C.sub.3)alkyl,
phenyl(C.sub.1-C.sub.4)a- lkyl,
phenylhydroxy(C.sub.1-C.sub.4)alkyl,
phenyl(C.sub.1-C.sub.4)alkoxy(C- .sub.1-C.sub.4)alkyl, thien-2- or
-3-yl(C.sub.1-C.sub.4)alkyl or fur-2- or
-3-yl(C.sub.1-C.sub.4)alkyl wherein said R.sub.4 rings are mono-,
di- or tri-substituted independently on carbon with H, halo,
(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy, trifluoromethyl,
hydroxy, amino or cyano; or
[0547] R.sub.4 is pyrid-2-, -3- or 4-yl(C.sub.1-C.sub.4)alkyl,
thiazol-2-, 4- or -5-yl(C.sub.1-C.sub.4)alkyl, imidazol -1-, -2-,
4- or -5-yl(C.sub.1-C.sub.4)alkyl, pyrrol-2- or
-3-yl(C.sub.1-C.sub.4)alkyl, oxazol-2-, -4- or
-5-yl-(C.sub.1-C.sub.4)alkyl, pyrazol-3-, 4- or
-5-yl(C.sub.1-C.sub.4)alkyl, isoxazol-3-, 4- or
-5-yl(C.sub.1-C.sub.4)alk- yl, isothiazol-3-, 4- or
-5-yl(C.sub.1-C.sub.4)alkyl, pyridazin-3- or
4-yl-(C.sub.1-C.sub.4)alkyl, pyrimidin-2-, 4-, -5- or
-6-yl(C.sub.1-C.sub.4)alkyl, pyrazin-2- or
-3-yl(C.sub.1-C.sub.4)alkyl or
1,3,5-triazin-2-yl(C.sub.1-C.sub.4)alkyl, wherein said preceding
R.sub.4 heterocycles are optionally mono- or di-substituted
independently with halo, trifluoromethyl, (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy, amino or hydroxy and said mono-or
di-substituents are bonded to carbon;
[0548] R.sub.5 is H, hydroxy, fluoro, (C.sub.1-C.sub.5)alkyl,
(C.sub.1-C.sub.5)alkoxy, (C.sub.1-C.sub.6)alkanoyl,
amino(C.sub.1-C.sub.4)alkoxy, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylami- no(C.sub.1-C.sub.4)alkoxy,
carboxy(C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.5)alkoxy-carbonyl(C.sub.1-C.sub.4)alkoxy,
benzyloxycarbonyl(C.sub.1-C.sub.4)alkoxy, or carbonyloxy wherein
said carbonyloxy is carbon-carbon linked with phenyl, thiazolyl,
imidazolyl, 1H-indolyl, furyl, pyrrolyl, oxazolyl, pyrazolyl,
isoxazolyl, isothiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl or
1,3,5-triazinyl and wherein said preceding R.sub.5 rings are
optionally mono-substituted with halo, (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy, hydroxy, amino or trifluoromethyl and said
mono-substituents are bonded to carbon;
[0549] R.sub.7 is H, fluoro or (C.sub.1-C.sub.5)alkyl; or
[0550] R.sub.5 and R.sub.7 can be taken together to be oxo;
[0551] R.sub.6 is carboxy, (C.sub.1-C.sub.8)alkoxycarbonyl,
C(O)NR.sub.8R.sub.9 or C(O)R.sub.12, wherein
[0552] R.sub.8 is H, (C.sub.1-C.sub.3)alkyl, hydroxy or
(C.sub.1-C.sub.3)alkoxy; and
[0553] R.sub.9 is H, (C.sub.1-C.sub.8)alkyl, hydroxy,
(C.sub.1C.sub.8) alkoxy,
methylene-perfluorinated(C.sub.1-C.sub.8)alkyl, phenyl, pyridyl,
thienyl, furyl, pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl,
imidazolyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, isoxazolyl,
isothiazolyl, pyranyl, piperidinyl, morpholinyl, pyridazinyl,
pyrimidinyl, pyrazinyl, piperazinyl or 1,3,5-triazinyl wherein said
preceding R.sub.9 rings are carbon-nitrogen linked; or
[0554] R.sub.9 is mono-, di- or tri-substituted
(C.sub.1-C.sub.5)alkyl, wherein said substituents are independently
H, hydroxy, amino, mono-N- or di-N,N-(C.sub.1-C.sub.5))alkylamino;
or
[0555] R.sub.9 is mono- or di-substituted (C.sub.1-C.sub.5)alkyl,
wherein said substituents are independently phenyl, pyridyl, furyl,
pyrrolyl, pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl,
pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, pyranyl,
pyridinyl, piperidinyl, morpholinyl, pyridazinyl, pyrimidinyl,
pyrazinyl, piperazinyl or 1,3,5-triazinyl
[0556] wherein the nonaromatic nitrogen-containing R.sub.9 rings
are optionally mono-substituted on nitrogen with
(C.sub.1-C.sub.6)alkyl, benzyl, benzoyl or
(C.sub.1-C.sub.6)alkoxycarbonyl and wherein the R.sub.9 rings are
optionally mono-substituted on carbon with halo,
(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy, hydroxy, amino, or
mono-N- and di-N,N (C.sub.1-C.sub.5)alkylamino provided that no
quaternized nitrogen is included and there are no nitrogen-oxygen,
nitrogen-nitrogen or nitrogen-halo bonds;
[0557] R.sub.12 is piperazin-1-yl,
4-(C.sub.1-C.sub.4)alkylpiperazin-1-yl, 4-formylpiperazin-1-yl,
morpholino, thiomorpholino, 1-oxothiomorpholino,
1,1-dioxo-thiomorpholino, thiazolidin-3-yl, 1-oxo-thiazolidin-3-yl,
1,1-dioxo-thiazolidin-3-yl,
2-(C.sub.1-C.sub.6)alkoxycarbonylpyrrolidin-1- -yl, oxazolidin-3-yl
or 2(R)-hydroxymethylpyrrolidin-1-yl; or
[0558] R.sub.12 is 3- and/or 4-mono- or di-substituted
oxazetidin-2-yl, 2-, 4-, and/or 5-mono- or di-substituted
oxazolidin-3-yl, 2-, 4-, and/or 5- mono- or di- substituted
thiazolidin-3-yl, 2-, 4-, and/or 5- mono- or di- substituted
1-oxothiazolidin-3-yl, 2-, 4-, and/or 5-mono- or di-substituted
1,1-dioxothiazolidin-3-yl, 3- and/or 4-, mono- or di-substituted
pyrrolidin-1-yl, 3-, 4- and/or 5-, mono-, di- or tri-substituted
piperidin-1-yl, 3-, 4-, and/or 5-mono-, di-, or tri-substituted
piperazin-1-yl, 3-substituted azetidin-1-yl, 4- and/or 5-, mono- or
di-substituted 1,2-oxazinan-2-yl, 3-and/or 4-mono- or
di-substituted pyrazolidin-1-yl, 4- and/or 5-, mono- or
di-substituted isoxazolidin-2-yl, 4- and/or 5-, mono- and/or
di-substituted isothiazolidin-2-yl wherein said R.sub.12
substituents are independently H, halo, (C.sub.1-C.sub.5)alkyl,
hydroxy, amino, mono-N- or di-N,N-(C.sub.1-C.sub.5)alkylamino,
formyl, oxo, hydroxyimino, (C.sub.1-C.sub.5)alkoxy, carboxy,
carbamoyl, mono-N-or di-N,N-(C.sub.1-C.sub.4)alkylcarbamoyl,
(C.sub.1-C.sub.4)alkoxyimino, (C.sub.1-C.sub.4)alkoxymethoxy,
(C.sub.1-C.sub.6)alkoxycarbonyl, carboxy(C.sub.1-C.sub.5)alkyl or
hydroxy(C.sub.1-C.sub.5)alkyl;
[0559] with the proviso that if R.sub.4 is H, methyl, ethyl or
n-propyl R.sub.5 is OH;
[0560] with the proviso that if R.sub.5 and R.sub.7 are H, then
R.sub.4 is not H, methyl, ethyl, n-propyl,
hydroxy(C.sub.1-C.sub.3)alkyl or
(C.sub.1-C.sub.3)alkoxy(C.sub.1-C.sub.3)alkyl and R.sub.6 is
C(O)NR.sub.8R.sub.9, C(O)R.sub.12 or
(C.sub.1-C.sub.4)alkoxycarbonyl.
[0561] Preferred glycogen phosphorylase inhibitors include
compounds having the Formula ID 17
[0562] and the pharmaceutically acceptable salts and prodrugs
thereof wherein
[0563] the dotted line ( - - - ) is an optional bond;
[0564] A is --C(H).dbd., --C((C.sub.1-C.sub.4)alkyl).dbd.,
--C(halo).dbd. or --N.dbd., when the dotted line ( - - - ) is a
bond, or A is methylene or --CH((C.sub.1-C.sub.4)alkyl)--, when the
dotted line ( - - - ) is not a bond;
[0565] R.sub.1, R.sub.10 or R.sub.11, are each independently H,
halo, cyano, 4-, 6-, or 7-nitro, (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy, fluoromethyl, difluoromethyl or
trifluoromethyl;
[0566] R.sub.2 is H;
[0567] R.sub.3 is H or (C.sub.1-C.sub.5)alkyl;
[0568] R.sub.4 is H, methyl, ethyl, n-propyl,
hydroxy(C.sub.1-C.sub.3)alky- l,
(C.sub.1-C.sub.3)alkoxy(C.sub.1-C.sub.3)alkyl,
phenyl(C.sub.1-C.sub.4)a- lkyl,
phenylhydroxy(C.sub.1-C.sub.4)alkyl,
(phenyl)((C.sub.1-C.sub.4)-alko- xy)(C.sub.1-C.sub.4)alkyl,
thien-2- or -3-yl(C.sub.1-C.sub.4)alkyl or fur-2- or
-3-yl(C.sub.1-C.sub.4)alkyl wherein said R.sub.4 rings are mono-,
di- or tri-substituted independently on carbon with H, halo,
(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy, trifluoromethyl,
hydroxy, amino, cyano or 4,5-dihydro-1H-imidazol-2-yl; or
[0569] R.sub.4 is pyrid-2-, -3- or -4-yl(C.sub.1-C.sub.4)alkyl,
thiazol-2-, -4- or -5-yl(C.sub.1-C.sub.4)alkyl, imidazol-2-, 4- or
-5-yl(C.sub.1-C.sub.4)alkyl, pyrrol-2- or
-3-yl(C.sub.1-C.sub.4)alkyl, oxazol-2-, -4- or
-5-yl(C.sub.1-C.sub.4)alkyl, pyrazol-3-, -4- or
-5-yl(C.sub.1-C.sub.4)alkyl, isoxazol-3-4- or
-5yl(C.sub.1-C.sub.4)alkyl, isothiazol-3-, -4- or
-5-yl(C.sub.1-C.sub.4)alkyl, pyridazin-3- or
-4-yl(C.sub.1-C.sub.4)alkyl, pyrimidin-2-, -4-, -5- or
-6-yl(C.sub.1-C.sub.4)alkyl, pyrazin-2- or
-3-yl(C.sub.1-C.sub.4)alkyl,
1,3,5-triazin-2-yl(C.sub.1-C.sub.4)alkyl or
indol-2-(C.sub.1-C.sub.4)alky- l, wherein said preceding R.sub.4
heterocycles are optionally mono- or di-substituted independently
with halo, trifluoromethyl, (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy, amino, hydroxy or cyano and said
substituents are bonded to carbon; or
[0570] R.sub.4 is R.sub.15-carbonyloxymethyl, wherein said R.sub.15
is phenyl, thiazolyl, imidazolyl, 1H-indolyl, furyl, pyrrolyl,
oxazolyl, pyrazolyl, isoxazolyl, isothiazolyl, pyridyl,
pyridazinyl, pyrimidinyl, pyrazinyl or 1,3,5-triazinyl and wherein
said preceding R.sub.15 rings are optionally mono- or
di-substituted independently with halo, amino, hydroxy,
(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy or trifluoromethyl
and said mono- or di-substituents are bonded to carbon;
[0571] R.sub.5 is H;
[0572] R.sub.6 is carboxy, (C.sub.1-C.sub.8)alkoxycarbonyl,
benzyloxycarbonyl, C(O)NR.sub.8R.sub.9 or C(O)R.sub.12 wherein
[0573] R.sub.8 is H, (C.sub.1-C.sub.6)alkyl,
cyclo(C.sub.3-C.sub.6)alkyl,
cyclo(C.sub.3-C.sub.6)alkyl(C.sub.1-C.sub.5)alkyl, hydroxy or
(C.sub.1-C.sub.8)alkoxy; and
[0574] R.sub.9 is H, cyclo(C.sub.3-C.sub.8)alkyl,
cyclo(C.sub.3-C.sub.8)al- kyl(C.sub.1-C.sub.5)alkyl,
cyclo(C.sub.4-C.sub.7)alkenyl,
cyclo(C.sub.3-C.sub.7)alkyl(C.sub.1-C.sub.5)alkoxy,
cyclo(C.sub.3-C.sub.7)alkyloxy, hydroxy,
methylene-perfluorinated(C.sub.1- -C.sub.8)alkyl, phenyl, or a
heterocycle wherein said heterocycle is pyridyl, furyl, pyrrolyl,
pyrrolidinyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl,
pyrazolinyl, pyrazolidinyl, isoxazolyl, isothiazolyl, pyranyl,
pyridinyl, piperidinyl, morpholinyl, pyridazinyl, pyrimidinyl,
pyrazinyl, piperazinyl, 1,3,5-triazinyl, benzothiazolyl,
benzoxazolyl, benzimidazolyl, thiochromanyl or
tetrahydrobenzothiazolyl wherein said heterocycle rings are
carbon-nitrogen linked; or
[0575] R.sub.9 is (C.sub.1-C.sub.6)alkyl or (C.sub.1-C.sub.8)alkoxy
wherein said (C.sub.1-C.sub.6)alkyl or (C.sub.1-C.sub.8)alkoxy is
optionally monosubstituted with cyclo(C.sub.4-C.sub.7)alken-1-yl,
phenyl, thienyl, pyridyl, furyl, pyrrolyl, pyrrolidinyl, oxazolyl,
thiazolyl, imidazolyl, pyrazolyl, pyrazolinyl, pyrazolidinyl,
isoxazolyl, isothiazolyl, pyranyl, piperidinyl, morpholinyl,
thiomorpholinyl, 1-oxothiomorpholinyl, 1,1-dioxothiomorpholinyl,
pyridazinyl, pyrimidinyl, pyrazinyl, piperazinyl, 1,3,5-triazinyl
or indolyl and wherein said (C.sub.1-C.sub.6)alkyl or
(C.sub.1-C.sub.8)alkoxy are optionally additionally independently
mono- or di-substituted with halo, hydroxy,
(C.sub.1-C.sub.5)alkoxy, amino, mono-N- or
di-N,N-(C.sub.1-C.sub.5)alkyla- mino, cyano, carboxy, or
(C.sub.1-C.sub.4)alkoxycarbonyl; and
[0576] wherein the R.sub.9 rings are optionally mono- or
di-substituted independently on carbon with halo,
(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy, hydroxy,
hydroxy(C.sub.1-C.sub.4)alkyl, amino(C.sub.1-C.sub.4)alkyl, mono-N-
or di-N,N-(C.sub.1-C.sub.4)alkylamin- o(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl, amino, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino, cyano, carboxy,
(C.sub.1-C.sub.5)alkoxycarbonyl, carbamoyl, formyl or
trifluoromethyl and said R.sub.9 rings may optionally be
additionally mono- or di-substituted independently with
(C.sub.1-C.sub.5)alkyl or halo;
[0577] with the proviso that no quaternized nitrogen on any R.sub.9
heterocycle is included;
[0578] R.sub.12 is morpholino, thiomorpholino, 1-oxothiomorpholino,
1,1-dioxothiomorpholino, thiazolidin-3-yl, 1-oxothiazolidin-3-yl,
1,1-dioxothiazolidin-3-yl, pyrrolidin-1-yl, piperidin-1-yl,
piperazin-1-yl, piperazin-4-yl, azetidin-1-yl, 1,2-oxazinan-2-yl,
pyrazolidin-1-yl, isoxazolidin-2-yl, isothiazolidin-2-yl,
1,2-oxazetidin-2-yl, oxazolidin-3-yl, 3,4-dihydroisoquinolin-2-yl,
1,3-dihydroisoindol-2-yl, 3,4-dihydro-2H-quinol-1-yl,
2,3-dihydro-benzo[1,4]oxazin-4-yl,
2,3-dihydro-benzo[1,4]-thiazine-4-yl,
3,4-dihydro-2H-quinoxalin-1-yl,
3,4-dihydro-benzo[c][1,2]oxazin-1-yl,
1,4-dihydro-benzo[d][1,2]oxazin-3-yl,
3,4-dihydro-benzo[e][1,2]-oxazin-2-- yl, 3H-benzo[d]isoxazol-2-yl,
3H-benzo[c]isoxazol-1-yl or azepan-1-yl,
[0579] wherein said R.sub.12 rings are optionally mono-, di- or
tri-substituted independently with halo, (C.sub.1-C.sub.5)alkyl,
(C.sub.1-C.sub.5)alkoxy, hydroxy, amino, mono-N- or
di-N,N-(C.sub.1-C.sub.5)alkylamino, formyl, carboxy, carbamoyl,
mono-N- or di-N,N-(C.sub.1-C.sub.5)alkylcarbamoyl,
(C.sub.1-C.sub.6)alkoxy(C.sub.- 1-C.sub.3)alkoxy,
(C.sub.1-C.sub.5)alkoxycarbonyl, benzyloxycarbonyl,
(C.sub.1-C.sub.5)alkoxycarbonyl(C.sub.1-C.sub.5)alkyl,
(C.sub.1-C.sub.4)alkoxycarbonylamino,
carboxy(C.sub.1-C.sub.5)alkyl, carbamoyl(C.sub.1-C.sub.5)alkyl,
mono-N- or di-N,N-(C.sub.1-C.sub.5)alkyl-
carbamoyl(C.sub.1-C.sub.5)alkyl, hydroxy(C.sub.1-C.sub.5)alkyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl,
amino(C.sub.1-C.sub.4)alky- l, mono-N- or
di-N,N-(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl, oxo,
hydroxyimino or (C.sub.1-C.sub.6)alkoxyimino and wherein no more
than two substituents are selected from oxo, hydroxyimino or
(C.sub.1-C.sub.6)alkoxyimino and oxo, hydroxyimino or
(C.sub.1-C.sub.6)alkoxyimino are on nonaromatic carbon; and
[0580] wherein said R.sub.12 rings are optionally additionally
mono- or di-substituted independently with (C.sub.1-C.sub.5)alkyl
or halo;
[0581] with the proviso that when R.sub.6 is
(C.sub.1-C.sub.5)alkoxycarbon- yl or benzyloxycarbonyl then R.sub.1
is 5-halo, 5-(C.sub.1-C.sub.4)alkyl or 5-cyano and R.sub.4 is
(phenyl)(hydroxy)(C.sub.1-C.sub.4)alkyl,
(phenyl)((C.sub.1-C.sub.4)alkoxy)(C.sub.1-C.sub.4)alkyl,
hydroxymethyl or Ar(C.sub.1-C.sub.2)alkyl, wherein Ar is thien-2-
or -3-yl, fur-2- or -3-yl or phenyl wherein said Ar is optionally
mono- or di-substituted independently with halo; with the provisos
that when R.sub.4 is benzyl and R.sub.5 is methyl, R.sub.12 is not
4-hydroxy-piperidin-1-yl or when R.sub.4 is benzyl and R.sub.5 is
methyl R.sub.6 is not C(O)N(CH.sub.3).sub.2;
[0582] with the proviso that when R.sub.1 and R.sub.10 and R.sub.11
are H, R.sub.4 is not imidazol-4-ylmethyl, 2-phenylethyl or
2-hydroxy-2-phenylethyl;
[0583] with the proviso that when both R.sub.8 and R.sub.9 are
n-pentyl, R.sub.1 is 5-chloro, 5-bromo, 5-cyano,
5(C.sub.1-C.sub.5)alkyl, 5(C.sub.1-C.sub.5)alkoxy or
trifluoromethyl;
[0584] with the proviso that when R.sub.12 is
3,4-dihydroisoquinol-2-yl, said 3,4-dihydroisoquinol-2-yl is not
substituted with carboxy((C.sub.1-C.sub.4)alkyl;
[0585] with the proviso that when R.sub.8 is H and R.sub.9 is
(C.sub.1-C.sub.6)alkyl, R.sub.9 is not substituted with carboxy or
(C.sub.1-C.sub.4)alkoxycarbonyl on the carbon which is attached to
the nitrogen atom N of NHR.sub.9; and
[0586] with the proviso that when R.sub.6 is carboxy and R.sub.1,
R.sub.10, R.sub.11 and R.sub.5 are all H, then R.sub.4 is not
benzyl, H, (phenyl)(hydroxy)methyl, methyl, ethyl or n-propyl.
[0587] In general an effective dosage for the pharmacological
combination compositions of this invention, for example the
ischemic damage reducing activities of combinations containing the
glycogen phosphorylase inhibitor compounds of this invention, is in
the range of 0.005 to 50 mg/kg/day, preferably 0.01 to 25 mg/kg/day
and most preferably 0.1 to 15 mg/kg/day.
[0588] Any glycogen phosphorylase inhibitor may be used as the
second compound of this invention. The term glycogen phosphorylase
inhibitor refers to any substance or agent or any combination of
substances and/or agents which reduces, retards, or eliminates the
enzymatic action of glycogen phosphorylase. The currently known
enzymatic action of glycogen phosphorylase is the degradation of
glycogen by catalysis of the reversible reaction of a glycogen
macromolecule and inorganic phosphate to glucose-1-phosphate and a
glycogen macromolecule which is one glucosyl residue shorter than
the original glycogen macromolecule (forward direction of
glycogenolysis). Such actions are readily determined by those
skilled in the art according to standard assays (e.g., as described
hereinafter). A variety of these compounds are included in the
following published international patent applications: PCT
application publication WO 96/39384 and WO96/39385. However, other
glycogen phosphorylase inhibitors will be known to those skilled in
the art.
[0589] Any sorbitol dehydrogenase inhibitor may be used as the
second compound of this invention. Such compounds inhibit the
formation of sorbitol dehydrogenase. Such actions are readily
determined by those skilled in the art according to standard assays
(e.g., as described hereinafter). A variety of these compounds will
be known to those skilled in the art (e.g., U.S. Pat. No.
5,728,704).
[0590] The compounds of the present invention pharmacologically
mimic the cardioprotective effects of ischemic preconditioning by
activating adenosine A-3 receptors and hence are useful as
therapeutic or prophylactic agents for diseases caused or
aggravated by ischemia or hypoxia, or ischemia/reperfusion for
example, cardiovascular diseases [e.g., arteriosclerosis,
arrhythmia (e.g. ischemic arrhythmia, arrhythmia due to myocardial
infarction, myocardial stunning, myocardial dysfunction, arrhythmia
after PTCA or after thrombolysis, etc.), angina pectoris, cardiac
hypertrophy, myocardial infarction, heart failure (e.g. congestive
heart failure, acute heart failure, cardiac hypertrophy, etc.),
restenosis after PTCA, PTCI, shock (e.g. hemorrhagic shock,
endotoxin shock, etc.)], renal diseases (e.g. diabetes mellitus,
diabetic nephropathy, ischemic acute renal failure, etc.) organ
disorders associated with ischemia or ischemic reperfusion [(e.g.
heart muscle ischemic reperfusion associated disorders, acute renal
failure, or disorders induced by surgical treatment such as
coronary artery bypass grafting (CABG) surgeries, vascular
surgeries, organ transplantation, non-cardiac surgeries or
percutaneous transluminal coronary angioplasty (PTCA)],
cerebrovascular diseases (e.g., ischemic stroke, hemorrhagic
stroke, etc.), cerebro ischemic disorders (e.g., disorders
associated with cerebral infarction, disorders caused after
cerebral apoplexy as sequelae, or cerebral edema. The compounds of
this invention can also be used as an agent for myocardial
protection during coronary artery bypass grafting (CABG) surgeries,
vascular surgeries, percutaneous transluminal coronary angioplasty
(PTCA), PTCI, organ transplantation, or non-cardiac surgeries.
[0591] Preferably, the compounds of this invention can be used as
agents for myocardial protection before, during, or after coronary
artery bypass grafting (CABG) surgeries, vascular surgeries,
percutaneous transluminal coronary angioplasty (PTCA), organ
transplantation, or non-cardiac surgeries.
[0592] Preferably, the compounds of this invention can be used as
agents for myocardial protection in patients presenting with
ongoing cardiac (acute coronary syndromes, e.g. myocardial
infarction or unstable angina) or cerebral ischemic events (e.g.,
stroke).
[0593] Preferably, the compounds of this invention can be used as
agents for chronic myocardial protection in patients with diagnosed
coronary heart disease (e.g. previous myocardial infarction or
unstable angina) or patients who are at high risk for myocardial
infarction (e.g., age greater than 65 and two or more risk factors
for coronary heart disease).
[0594] Accordingly, the compounds of this invention reduce
mortality.
[0595] The utility of the compounds of the present invention as
medical agents in the treatment of diseases, such as are detailed
herein in mammals (e.g., humans), for example, myocardial
protection during surgery or mycardial protection in patients
presenting with ongoing cardiac or cerebral ischemic or hypoxic
events or chronic cardioprotection in patients with diagnosed
coronary heart disease, or at risk for coronary heart disease,
cardiac dysfunction or myocardial stunning is demonstrated by the
activity of the compounds of this invention in conventional
preclinical cardioprotection assays [see the in vivo assay in
Klein, H. et al., Circulation 92:912-917 (1995); the isolated heart
assay in Tracey, W. R. et al., Cardiovascular Research 33:410415
(1997); the antiarrhythmic assay in Yasutake M. et al., Am. J.
Physiol., 36:H2430-H2440 (1994); the NMR assay in Kolke et al., J.
Thorac. Cardiovasc. Surg. 112: 765-775 (1996)] and the additional
in vitro and in vivo assays described below. Such assays also
provide a means whereby the activities of the compounds of this
invention can be compared with the activities of other known
compounds. The results of these comparisons are useful for
determining dosage levels in mammals, including humans, for the
treatment of such diseases.
Human Adenosine A.sub.1 and A.sub.3 Receptor Assays
[0596] Materials
[0597] Full-length human adenosine A.sub.1 and A.sub.3 receptor
cDNA's subcloned into the eukaryotic expression vector pRcCMV
(Invitrogen) were purchased from The Garvan Institute, Sydney,
Australia. Chinese hamster ovary (CHO-K1) cells were obtained from
the American Type Tissue Culture Collection (Rockville, Md., USA).
DMEM and DMEM/F12 culture media and foetal calf serum were obtained
from Gibco-BRL (Grand Island, N.Y., USA). The A1/A3 adenosine
receptor agonist N6-(4-amino-3-[1251]iodobenzyl)adeno- sine
(.sup.125I-ABA) was prepared by New England Nuclear (Boston, Mass.,
USA). Adenosine deaminase (ADA) was obtained from Boehringer
Mannheim (Indianapolis, Ind., USA). The phosphodiesterase inhibitor
RO-20-1724 was obtained from Research Biochemicals International
(Matick, Mass., USA).
Expression of Human Adenosine A1 and A3 Receptors
[0598] For stable expression studies adenosine receptor A.sub.1 and
A.sub.3 expression plasmids (20 .mu.g) are transfected into CHO-K1
cells, or HEK 293s cells, respectively, grown in DMEM/F12 (CHO) or
DMEM (HEK 293s), with 10% foetal calf serum media, using a calcium
phosphate mammalian cell transfection kit (5 Prime-3 Prime). Stable
transfectants are obtained by selection in complete media
containing 500 .mu.g/ml (CHO) or 700 .mu.g/ml (HEK 293s) active
neomycin (G418) and screened for expression by [.sup.125I]-ABA
binding.
Receptor Membrane Preparation
[0599] Cells stably expressing either human A.sub.1 or human
A.sub.3 receptors are collected by centrifugation at 300.times.g
for 5 minutes, the supernatant is discarded and the cell pellet is
resuspended in cell buffer consisting of (mmoles/L): HEPES (10),
MgCl.sub.2 (5), PMSF (0.1), bacitracin (100 .mu.g/ml), leupeptin
(10 .mu.g/ml), DNAse I (100 .mu.g/ml), ADA (2 U/ml), pH 7.4. Crude
cell membranes are prepared by repeated aspiration through a 21
gauge needle, collected by centrifugation at 60,000.times.g for 10
minutes and stored in cell buffer at -80.degree. C.
Estimation of Compound Binding Affinity Constants (K.sub.i)
[0600] Receptor membranes are resuspended in incubation buffer
consisting of (mmoles/L): HEPES (10), EDTA (1), MgCl.sub.2 (5), pH
7.4. Binding reactions (10-20 .mu.g membrane protein) are carried
out for one hour at room temperature in 250 .mu.l incubation buffer
containing 0.1 nM of .sup.125I-ABA (2200 Ci/mmol) and increasing
concentrations of compound (0.1 nM-30 .mu.M). The reaction is
stopped by rapid filtration with ice-cold PBS, through glass fibre
filters (presoaked in 0.6% polyethylenimine) using a Tomtec 96-well
harvester (Orange, Conn., USA). Filters are counted in a Wallac
Microbeta liquid scintillation counter (Gaithersberg, Md., USA).
Nonspecific binding is determined in the presence of 5 .mu.M I-ABA.
Compound inhibitory constants (K.sub.i) are calculated by fitting
binding data via nonlinear least squares regression analysis to the
equation: % Inhibition=100/[1+(10.sup.C/10.sup.X).sup.D], where
X=log [drug concentration], C (IC.sub.50)=log [drug concentration
at 50% inhibtion], and D=the Hill slope. At the concentration of
radioligand used in the present study (10 fold<K.sub.D),
IC.sub.50=K.sub.i.
Assessment of Human Adenosine A3 Receptor Agonist Activity
[0601] Adenosine A3 agonist activity is assessed by compound
inhibition of isoproternol-stimulated cAMP levels. HEK293s cells
stably transfected with human A3 receptors (as described above) are
washed with Phosphate Buffered Saline (PBS) (Ca/Mg-free) and
detached with 1.0 mM EDTA/PBS. Cells are collected by
centrifugation at 300.times.g for 5 minutes and the supernatant
discarded. The cell pellet is dispersed and resuspended in cell
buffer (DMEM/F12 containing 10 mM HEPES, 20 .mu.M RO-20-1724 and 1
U/ml ADA). Following preincubation of cells (100,000/well) for 10
min at 37.degree. C., 1 .mu.M isoproterenol, with or without
increasing concentrations (0.1 nM-300 nM) test compound, and the
incubation is continued for 10 min. Reactions are terminated by the
addition of 1.0 N HCl followed by centrifugation at 2000.times.g
for 10 minutes. Sample supernatants (10 .mu.l) are removed and cAMP
levels determined by radioimmunoassay (New England Nuclear, Boston,
Mass., USA). The basal and control isoproterenol-stimulated cAMP
accumulation (pmol/ml/100,000 cells) are routinely 3 and 80,
respectively. Smooth curves are fitted to the data via nonlinear
least squares regression analysis to the equation: %
isoproterenol-stimulated
cAMP=100/[1+(.sub.10.sup.X/10.sup.C).sup.D], where X=log [drug
concentration], C (IC.sub.50)=log [drug concentration at 50%
inhibition], and D=the Hill slope.
[0602] As background information, it is noted that brief periods of
myocardial ischemia followed by coronary artery reperfusion
protects the heart from subsequent severe myocardial ischemia
(Murry et al., Circulation 74:1124-1136, 1986).
[0603] The therapeutic effects of the compounds of this invention
in preventing heart tissue damage resulting from an ischemic insult
can be demonstrated in vitro along lines presented in Tracey et al.
(Cardiovasc. Res., 33:410415, 1997), as described specifically
herein. Cardioprotection, as indicated by a reduction in infarcted
myocardium, can be induced pharmacologically using adenosine
receptor agonists in isolated, retrogradely perfused rabbit hearts
as an in vitro model of myocardial ischemic preconditioning (Tracey
et al. (Cardiovasc. Res., 33:410-415, 1997)). The in vitro test
described below demonstrates that a test compound (i.e., a compound
as claimed herein) can also pharmacologically induce
cardioprotection, i.e., reduced myocardial infarct size, when
administered to a rabbit isolated heart. The effects of the test
compound are compared to ischemic preconditioning. The exact
methodology is described below.
[0604] The protocol used for these experiments closely follows that
described by Tracey et al. (Cardiovasc. Res., 33:410415, 1997).
Male New Zealand White rabbits (3-4 kg) are anesthetized with
sodium pentobarbital (30 mg/kg, i.v.). After deep anesthesia is
achieved (determined by the absence of an ocular blink reflex) the
animal is intubated and ventilated with 100% O.sub.2 using a
positive pressure ventilator. A left thoracotomy is performed, the
heart exposed, and a snare (2-0 silk) is placed loosely around a
prominent branch of the left coronary artery, approximately 2/3 of
the distance from the apex of the heart. The heart is removed from
the chest and rapidly (<30 sec) mounted on a Langendorff
apparatus. The heart is retrogradely perfused in a
non-recirculating manner with a modified Krebs solution (NaCl 118.5
mM, KCl 4.7 mM, Mg SO.sub.4 1.2 mM, KH.sub.2PO.sub.4 1.2 mM,
NaHCO.sub.3 24.8 mM, CaCl.sub.2 2.5 mM, and glucose 10 mM), at a
constant pressure of 80 mmHg and a temperature of 37.degree. C.
Perfusate pH is maintained at 7.4-7.5 by bubbling with 95%
O.sub.2/5% CO.sub.2. Heart temperature is tightly controlled by
using heated reservoirs for the physiological solution and water
jacketing around both the perfusion tubing and the isolated heart.
Heart rate and left ventricular pressures are determined via a
latex balloon which is inserted in the left ventricle and connected
by stainless steel tubing to a pressure transducer. The
intraventricular balloon is inflated to provide a systolic pressure
of 80-100 mmHg, and a diastolic pressure <10 mmHg. Total
coronary flow is also continuously monitored using an in-line flow
probe and normalized for heart weight.
[0605] The heart is allowed to equilibrate for 30 min, over which
time the heart must show stable left ventricular pressures within
the parameters outlined above. If the heart rate falls below 180
bpm at any time prior to the 30 min period of regional ischemia,
the heart is paced at about 200 bpm for the remainder of the
experiment. Ischemic preconditioning is induced by total cessation
of cardiac perfusion (global ischemia) for 5 min, followed by
reperfusion for 10 min. The regional ischemia is provided by
tightening the snare around the coronary artery branch. Following
the 30 min regional ischemia, the snare is released and the heart
reperfused for an additional 120 min.
[0606] Pharmacological cardioprotection is induced by infusing the
test compound at predetermined concentrations, for a 5 min period
which ends 10 min before the 30 min regional ischemia. Hearts which
receive test compounds do not undergo the period of ischemic
preconditioning.
[0607] At the end of the 120 min reperfusion period, the coronary
artery snare is tightened, and a 0.5% suspension of fluorescent
zinc cadmium sulfate particles (1-10 .mu.M) Duke Scientific
Corp.(Palo Alto, Calif.) is perfused through the heart; this stains
all of the myocardium, except that area-at-risk for infarct
development (area-at-risk). The heart is removed from the
Langendorff apparatus, blotted dry, wrapped in aluminum foil and
stored overnight at -20.degree. C. The next day, the heart is
sliced into 2 mm transverse sections from the apex to the top of
the ventricles. The slices are stained with 1% triphenyl
tetrazolium chloride (TTC) in phosphate-buffered saline for 20 min
at 37.degree. C. Since TTC reacts with living tissue (containing
NAD-dependent dehydrogenases), this stain differentiates between
living (red stained) tissue, and dead tissue (unstained infarcted
tissue). The infarcted area (no stain) and the area-at-risk (no
fluorescent particles) are calculated for each slice of left
ventricle using a precalibrated image analyzer. To normalize the
ischemic injury for differences in the area-at-risk between hearts,
the data is expressed as the ratio of infarct area vs. area-at-risk
(% IA/AAR). All data are expressed as mean.+-.SE and compared
statistically using a Mann-Whitney non-parametric test with a
Bonferroni correction for multiple comparisons. Significance is
considered as p<0.05.
[0608] The results from the above in vitro test demonstrate that
compounds of this invention induce significant cardioprotection
relative to the control group.
[0609] The therapeutic effects of the compounds of this invention
in preventing heart tissue damage resulting from an ischemic insult
can also be demonstrated in vivo along lines presented in Liu et
al. (Circulation, Vol. 84:350-356, 1991) as described specifically
herein. The in vivo assay tests the cardioprotection of the test
compound relative to the control group which receives saline
vehicle. Cardioprotection, as indicated by a reduction in infarcted
myocardium, can be induced pharmacologically using intravenously
administered adenosine receptor agonists in intact, anesthetized
rabbits studied as an in vivo model of myocardial ischemic
preconditioning (Liu et al., Circulation 84:350-356, 1991). The in
vivo assay tests whether compounds can pharmacologically induce
cardioprotection, i.e., reduced myocardial infarct size, when
parenterally administered to intact, anesthetized rabbits. The
effects of the compounds of this invention can be compared to
ischemic preconditioning. The methodology is described below.
[0610] Surgery: New Zealand White male rabbits (3-4 kg) are
anesthetized with sodium pentobarbital as a bolus dose (30 mg/kg,
i.v.) followed by an infusion (100 mg/kg/hr, i.v.) to maintain a
surgical plane of anesthesia. A tracheotomy is performed via a
ventral midline cervical incision and the rabbits are ventilated
with 100% oxygen using a positive pressure ventilator. The
ventilation is adjusted to maintain pH and PCO.sub.2 within
physiological ranges. Body temperature is held constant at
38.degree. C. using a heating pad. Catheters are placed in the left
jugular vein for drug administration and in the left carotid artery
for blood pressure measurements. The hearts are then exposed
through a left thoracotomy and a snare (00 silk) placed around a
prominent branch of the left coronary artery approximately
two-thirds of the distance from the apex of the heart. Ischemia is
induced by pulling the snare tight. Releasing the snare allows the
ischemic area to reperfuse. Myocardial ischemia is evidenced by
regional cyanosis; reperfusion is evidenced by reactive
hyperemia.
[0611] Protocol: Once arterial pressure and heart rate have been
stable for at least 120 minutes the test is started. Ischemic
preconditioning is induced by occluding the coronary artery for 5
min followed by a 10 min reperfusion. Pharmacological
preconditioning is induced by infusing test compound over, for
example, 5 minutes and allowing 10 minutes before further
intervention. Following ischemic preconditioning, pharmacological
preconditioning or no conditioning (unconditioned, vehicle control)
the artery is occluded for 30 minutes and then reperfused for two
hours to induce myocardial infarction.
[0612] At the end of the 2 hour reperfusion period, the hearts are
quickly removed, placed on a Langendorff apparatus, and perfused
for 1 minute with normal saline heated to body temperature
(38.degree. C.). The silk suture used as the snare is then tied
tightly to reocclude the artery and a 0.5% suspension of
fluorescent zinc cadmium sulphate particles (1-10 82 m) Duke
Scientific Corp. (Palo Alto, Calif.) is infused with the perfusate
to stain all of the myocardium except for the area at risk
(nonfluorescent ventricle). The hearts are then removed from the
apparatus, blotted dry, wrapped in aluminum foil and stored
overnight at -20.degree. C. On the following day, the ventricles
are sliced into 2 mm transverse slices sections from apex to base
and stained with 1% triphenyl tetrazolium chloride (TTC) in
phosphate buffered saline for 20 minutes at 38.degree. C. Since TTC
reacts with living tissue (NAD-dependent dehydrogenase present),
this stain differentiates between living (red stained) tissue, and
dead tissue (unstained infarcted tissue). The infarcted area (no
stain) and the area at risk (no fluorescent particles) are
calculated for each slice of left ventricle using a pre-calibrated
image analyzer. To normalize the ischemic injury for differences in
the area at risk between hearts, the data is expressed as the ratio
of infarct area vs. area at risk (% IA/MR). All data are expressed
as Mean.+-.SEM and compared statistically using single factor ANOVA
or Mann Whitney non parametric test. Significance is considered as
p<0.05.
[0613] The compounds of this invention can be tested for their
utility in reducing or preventing ischemic or hypoxic injury in
non-cardiac tissues, for example, the brain, or the liver,
utilizing procedures reported in the scientific literature. The
compounds of this invention in such tests can be administered by
the preferred route and vehicle of administration and at the
preferred time of administration either prior to the ischemic
episode, during the ischemic or hypoxic episode, following the
ischemic or hypoxic episode (reperfusion period) or during any of
the below-mentionedexperimental stages.
[0614] The benefit of the invention to reduce ischemic or hypoxic
brain damage can be demonstrated, for example, in mammals using the
method of Park, et al, (Ann. Neurol. 1988;24:543-551). According to
the procedure of Park, et al., adult male Sprague Dawley rats are
anesthetized initially with 2% halothane, and thereafter by
mechanical ventilation with a nitrous oxide-oxygen mixture
(70%:30%) containing 0.5-1% halothane. A tracheostomy is then
performed. The stroke volume of the ventilator is adjusted to
maintain arterial carbon dioxide tension at approximately 35 mm Hg
and adequate arterial oxygenation (PaO.sub.2>90 mm Hg). Body
temperature can be monitored by a rectal thermometer, and the
animals can be maintained normothermic, if necessary, by external
heating. The animals next undergo subtemporal craniectomy to expose
the main trunk of the left middle cerebral artery (MCA) under an
operating microscope, and the exposed artery is occluded with
microbipolar coagulation to generate large ischemic lesions in the
cerebral cortex and basal ganglia. After three hours of MCA
occlusion, the rats are deeply anesthetized with 2% halothane and a
thoracotomy is performed to infuse heparinized saline into the left
ventricle. The effluent is collected via an incision of the right
atrium. The saline washout is followed by approximately 200 ml of a
40% formaldehyde, glacial acetic acid and absolute methanol
solution (FAM; 1:1:8, v/v/v), then the animals are decapitated and
the head is stored in fixative for 24 hours. The brain is then
removed, dissected, embedded in paraffin wax, and sectioned
(approximately 100 sections of 0.2 mm per brain). The sections are
then stained with hematoxylin-eosin or with a combination of cresyl
violet and Luxol.RTM. fast blue, and examined by light microscopy
to identify and quantitate the ischemic damage using a
precalibrated image analyzer. The ischemic volumes and areas are
expressed in absolute units (mm.sup.3 and mm.sup.2) and as a
percentage of the total region examined. The effect of the
compounds, compositions and methods of this invention to reduce
ischemic brain damage induced by MCA occlusion is noted based on a
reduction in the area or volume of relative or absolute ischemic
damage in the brain sections from the rats in the treatment group
compared to brain sections from rats in a placebo-treated control
group.
[0615] Other methods which could alternatively be utilized to
demonstrate the benefit of the invention to reduce ischemic or
hypoxic brain damage include those described by Nakayama, et al. in
Neurology 1988,38:1667-1673; Memezawa, et al. in Stroke
1992,23:552-559; Folbergrova, et al. in Proc. Natl. Acad. Sci
1995,92:5057-5059; and Gotti, et al. in Brain Res.
1990,522:290-307.
[0616] The benefit of the compounds, compositions and methods of
this invention to reduce ischemic or hypoxic liver damage can be
demonstrated, for example, in mammals using the method of Yokoyama,
et al. (Am. J. Physiol. 1990;258;G564-G570). According to the
procedure of Yokoyama, et al., fasted adult male Sprague Dawley
rats are anesthetized with sodium pentobarbital (40 mg/kg i.p.),
then the animals are tracheotomized and mechanically ventilated
with room air. The liver is extirpated and placed in an
environmental chamber maintained at constant temperature
(37.degree. C.), then perfused through the portal vein at a
constant pressure of 15 cm H.sub.2O with a modified,
hemoglobin-free Krebs-Henseleit buffer (in mM: 118 NaCl, 4.7 KCl,
27 NaHCO.sub.3, 2.5 CaCl.sub.2, 1.2 MgSO.sub.4, 1.2
KH.sub.2PO.sub.4, 0.05 EDTA, and 11 mM glucose, plus 300 U
heparin). The pH of the perfusate is maintained at 7.4 by gassing
the buffer with 95% O.sub.2-5% CO.sub.2. Each liver is perfused at
a flow rate of 20 ml/min in a single-pass manner for a 30 min
washout and equilibration period (preischemic period), followed by
a 2 hour period of global ischemia, and then a 2 hour period of
reperfusion under conditions identical to the preischemic period.
Aliquots (20 ml) of the perfusate are collected during the
preischemic period, immediately after the occlusive ischemic
period, and every 30 min of the 2 hour reperfusion period. The
perfusate samples are assayed for the appearance of hepatocellular
enzymes, for example, aspartate amino-transferase (AST), alanine
amino-transferase (ALT), and lactate dehydrogenase (LDH), which are
taken to quantitatively reflect the degree of ischemic liver tissue
damage during the procedure. AST, ALT, and LDH activities in the
perfusate can be determined by several methods, for example, by the
reflectometry method using an automatic Kodak Ektachem 500 analyzer
reported by Nakano, et al. (Hepatology 1995;22:539-545). The effect
of the compounds, compositions and methods of this invention in
reducing ischemic liver damage induced by occlusion is noted based
on a reduction in the release of hepatocellular enzymes immediately
following the occlusive period and/or during the
postischemic-reperfusion period in the perfused livers from the
rats in the treatment group compared to perfused livers from rats
in a placebo-treated control group.
[0617] Other methods and parameters which could alternatively be
utilized to demonstrate the benefit of the compounds, compositions
and methods of this invention in reducing ischemic or hypoxic liver
damage include those described by Nakano, et al. (Hepatology
1995;22:539-545).
Measurement of Human NHE-1 Inhibitory Activity
[0618] Methodologies for measurement of human NHE-1 activity and
inhibitor potency are based on those published by Watson et al.,
Am. J. Physiol., 24:G229-G238, 1991), where NHE-mediated recovery
of intracellular pH is measured following intracellular
acidification. Thus, fibroblasts stably expressing human NHE-1
(Counillon, L. et al., Mol. Pharmacol., 44:1041-1045 (1993) are
plated onto collagen coated 96 well plates (50,000/well) and grown
to confluence in growth media (DMEM high glucose, 10% fetal bovine
serum, 50 .mu./ml penicillin and streptomycin). Confluent plates
are incubated for 30 min at 37.degree. C. with the pH sensitive
fluorescent probe BCECF (5 .mu.M; Molecular Probes, Eugene, Oreg.).
BCECF loaded cells are incubated for 30 min at 37.degree. C. in
acid loading media (70 mM choline chloride, 50 mM NHCl.sub.4, 5 mM
KCl, 1 mM MgCl.sub.2, 1.8 mM CaCl.sub.2, 5 mM glucose, 10 mM HEPES,
pH 7.5), and then placed in a Fluorescent Imaging Plate Reader
(Molecular Devices, Calif.). BCECF fluorescence is monitored using
excitation and emission wavelengths of 485 nM and 525 nM,
respectively. Intracellular acidification is initiated via rapid
replacement of acid loading media with recovery media (120 mM NaCl,
5 mM KCl, 1 mM MgCl.sub.2, 1.8 mM CaCl.sub.2, 5 mM glucose, 10 mM
HEPES, pH 7.5) .+-.test compound, and NHE-mediated recovery of
intracellular pH is monitored as the subsequent time-dependent
increase BCECF fluorescence. The potency of human NHE-1 inhibitors
is calculated as the concentration that reduces recovery of
intracellular pH by 50% (IC.sub.50). Under these conditions
reference NHE inhibitors amiloride and HOE-642 had IC.sub.50 values
for human NHE-1 of 50 .mu.M and 0.5 .mu.M, respectively.
Aldose Reductase Inhibitor Assays
[0619] Male Sprague-Dawley rats are rendered diabetic by injection
of streptozocin at 55 mg/kg, i.v., in pH 4.5 citrate buffer. They
are fed ad libitum in controlled conditions of housing, temperature
and lighting. After five weeks of diabetes, the rats are
anesthetized with an overdose of pentobarbital, and tissues are
rapidly removed and analyzed for sorbitol and fructose.
[0620] Sorbitol levels are analyzed according to the method of
Donald M. Eades et al., "Rapid Analysis of Sorbitol, Galactitol,
Mannitol and Myoinositol Mixtures From Biological Sources", Journal
of Chromatography, 490, 1-8, (1989).
[0621] Fructose in rat tissues is enzymatically measured using a
modification of the method of Ameyama (Methods in Enzymology,
89:20-29,1982), in which ferricyanide was replaced by resazurin, a
dye that is reduced to the highly fluorescent resorufin. The amount
of resorufin fluorescence is stoichiometric with the amount of
fructose oxidized by fructose dehydrogenase. The assay contains 0.1
ml neutralized 6% perchloric acid nerve extract in a final volume
of 1.5 ml. Following incubation for 60 minutes at room temperature
in a closed drawer, sample fluorescence is determined at
excitation=560 nm, emission=580 nm with slits of 5 mm each in a
Perkin-Elmer model 650-40 fluorescence spectrophotometer. Fructose
concentrations are calculated by comparison with a series of known
fructose standards.
Measurement of SDH Activity
[0622] Male Sprague-Dawley rats (350-400 g) are used for these
experiments. Diabetes is induced in some of the rats by a tail vein
injection of streptozocin, 85 mg/kg. Twenty-four hours later, 4
groups of diabetic rats are given a single dose of the test
compound of formula I of this invention (0.001 to 100 mg/kg) by
oral gavage. Animals are sacrificed 4-6 hours after.dosing and
blood and sciatic nerves are harvested. Tissues and cells are
extracted with 6% perchloric acid.
[0623] Sorbitol in erythrocytes and nerves is measured by a
modification of the method of R. S. Clements et al. (Science, 166:
1007-8, 1969). Aliquots of tissue extracts are added to an assay
system which has final concentrations of reagents of 0.033 M
glycine, pH 9.4, 800 mM .beta.-nicotine adenine dinucleotide, and 4
units/ml of sorbitol dehydrogenase. After incubation for 30 minutes
at room temperature, sample fluorescence is determined on a
fluorescence spectrophotometer with excitation at 366 nm and
emission at 452 nm. After subtracting appropriate blanks, the
amount of sorbitol in each sample is determined from a linear
regression of sorbitol standards processed in the same manner as
the tissue extracts.
[0624] Fructose is determined by a modification of the method
described by M. Ameyama, Methods in Enzymology, 89: 20-25 (1982).
Resazurin is substituted for ferricyanide. Aliquots of tissue
extracts are added to the assay system, which has final
concentrations of reagents of 1.2 M citric acid, pH 4.5, 13 mM
resazurin, 3.3 units/ml of fructose dehydrogenase and 0.068% Triton
X-100. After incubation for 60 minutes at room temperature, sample
fluorescence is determined on a fluorescence spectrophotometer with
excitation at 560 nm and emission at 580 nm. After subtracting
appropriate blanks, the amount of fructose in each sample is
determined from a linear regression of fructose standards processed
in the same manner as the tissue extracts.
[0625] SDH activity is measured by a modification of the method
described by U. Gerlach, Methodology of Enzymatic Analyses, edited
by H. U. Bergmeyer, 3, 112-117 (1983). Aliquots of sera or urine
are added to the assay system, which has final concentrations of
reagents of 0.1 M potassium phosphate buffer, pH 7.4, 5 mM NAD, 20
mM sorbitol, and 0.7 units/ml of sorbitol dehydrogenase. After
incubation for 10 minutes at room temperature, the average change
in sample absorbance is determined at 340 nm. SDH activity was
presented as milliOD.sub.340 units/minute (OD.sub.340=optical
density at 340 nm).
Glycogen Phosphorylase Inhibitor Assays
[0626] The three different purified glycogen phosphorylase (GP)
isoenzymes, wherein glycogen phosphorylase is in the activated "a"
state (referred to as glycogen phosphorylase a, or the abbreviation
GPa), and referred to here as human liver glycogen phosphorylase a
(HLGPa), human muscle glycogen phosphorylase a (HMGPa), and human
brain glycogen phosphorylase a (HBGPa), can be obtained by the
following procedures.
[0627] Expression and fermentation
[0628] The HLGP and HMGP cDNAs are expressed from plasmid pKK233-2
(Pharmacia Biotech. Inc., Piscataway, N.J.) in E. coli strain XL-1
Blue (Stratagene Cloning Systems, LaJolla, Calif.). The strain is
inoculated into LB medium (consisting of 10 g tryptone, 5 g yeast
extract, 5 g NaCl, and 1 ml 1N NaOH per liter) plus 100 mg/L
ampicillin, 100 mg/L pyridoxine and 600 mg/L MnCl.sub.2 and grown
at 37.degree. C. to a cell density of OD.sub.550=1.0. At this
point, the cells are induced with 1 mM
isopropyl-1-thio-.beta.-D-galactoside (IPTG). Three hours after
induction the cells are harvested by centrifugation and cell
pellets are frozen at -70.degree. C. until needed for
purification.
[0629] The HBGP cDNA can be expressed by several methodologies, for
example, by the method described by Crerar, et al. (J. Biol. Chem.
270:13748-13756). The method described by Crerar, et al. (J. Biol.
Chem. 270:13748-13756) for the expression of HBGP is as follows:
the HBGP cDNA can be expressed from plasmid pTACTAC in E. Coli
strain 25A6. The strain is inoculated into LB medium (consisting of
10 g tryptone, 5 g yeast extract, 5 g NaCl, and 1 ml 1N NaOH per
liter) plus 50 mg/L ampicillin and grown overnight, then
resuspended in fresh LB medium plus 50 mg/L ampicillin, and
reinoculated into a 40.times.volume of LB/amp media containing 250
.mu.M isopropyl-1-thio-.beta.-D-galactoside (IPTG), 0.5 mM
pyridoxine and 3 mM MgCl.sub.2 and grown at 22.degree. C. for 48-50
hours. The cells can then be harvested by centrifugation and cell
pellets are frozen at -70.degree. C. until needed for
purification.
[0630] The HLGP cDNA is expressed from plasmid pBlueBac III
(Invitrogen Corp., San Diego, Calif.) which is cotransfected with
BaculoGold Linear Viral DNA (Pharmingen, San Diego, Calif.) into
Sf9 cells. Recombinant virus is subsequently plaque-purified. For
production of protein, Sf9 cells grown in serum-free medium are
infected at a multiplicity of infection (moi) of 0.5 and at a cell
density of 2.times.10.sup.6 cells/ml. After growth for 72 hours at
27.degree. C., cells are centrifuged, and the cell pellets frozen
at -70.degree. C. until needed for purification. Purification of
Glycogen Phosphorylase expressed in E. coli
[0631] The E. coli cells in pellets described above are resuspended
in 25 mM .beta.-glycerophosphate (pH 7.0) with 0.2 mM DTT, 1 mM
MgCl.sub.2, plus the following protease inhibitors:
1 0.7 .mu.g/mL Pepstatin A 0.5 .mu.g/mL Leupeptin 0.2 mM
phenylmethylsulfonyl fluoride (PMSF), and 0.5 mM EDTA,
[0632] lysed by pretreatment with 200 .mu.g/mL lysozyme and 3
.mu.g/mL DNAase followed by sonication in 250 mL batches for
5.times.1.5 minutes on ice using a Branson Model 450 ultrasonic
cell disrupter (Branson Sonic Power Co., Danbury Conn.). The E.
coli cell lysates are then cleared by centrifugation at
35,000.times.g for one hour followed by filtration through 0.45
micron filters. GP in the soluble fraction of the lysates
(estimated to be less than 1% of the total protein) is purified by
monitoring the enzyme activity (as described in GPa Activity Assay
section, below) from a series of chromatographic steps detailed
below.
[0633] Immobilized Metal Affinity Chromatography (IMAC)
[0634] This step is based on the method of Luong et al (Luong et
al. Journal of Chromatography (1992) 584, 77-84.). 500 mL of the
filtered soluble fraction of cell lysates (prepared from
approximately 160-250 g of original cell pellet) are loaded onto a
130 mL column of IMAC Chelating-Sepharose (Pharmacia LKB
Biotechnology, Piscataway, N.J.) which has been charged with 50 mM
CuCl.sub.2 and 25 mM .beta.-glycerophosphate, 250 mM NaCl and 1 mM
imidazole at pH 7 equilibration buffer. The column is washed with
equilibration buffer until the A.sub.280 returns to baseline. The
sample is then eluted from the column with the same buffer
containing 100 mM imidazole to remove the bound GP and other bound
proteins. Fractions containing the GP activity are pooled
(approximately 600 mL), and ethylenediaminetetraacetic acid (EDTA),
DL-dithiothreitol (DTT), phenylmethylsulfonyl fluoride (PMSF),
leupeptin and pepstatin A are added to obtain 0.3 mM, 0.2 mM, 0.2
mM, 0.5 .mu.g/mL and 0.7 .mu.g/mL concentrations respectively. The
pooled GP is desalted over a Sephadex G-25 column (Sigma Chemical
Co., St. Louis, Mo.) equilibrated with 25 mM Tris-HCl (pH 7.3), 3
mM DTT buffer (Buffer A) to remove imidazole and is stored on ice
until the second chromatographic step.
[0635] 5'-AMP-Sepharose Chromatography
[0636] The desalted pooled GP sample (approximately 600 mL) is next
mixed with 70 mL of 5'-AMP Sepharose (Pharmacia LKB Biotechnology,
Piscataway, N.J.) which has been equilibrated with Buffer A (see
above). The mixture is gently agitated for one hour at 22.degree.
C. then packed into a column and washed with Buffer A until the
A.sub.208 returns to baseline. GP and other proteins are eluted
from the column with 25 mM Tris-HCl, 0.2 mM DTT and 10 mM adenosine
5'-monophosphate (AMP) at pH 7.3 (Buffer B). GP-containing
fractions are pooled following identification by determining enzyme
activity (described below) and visualizing the M.sub.r
approximately 97 kdal GP protein band by sodium dodecyl sulfate
polyacrylamide gel electrophoresis (SDS-PAGE) followed by silver
staining (2D-silver Stain II "Daiichi Kit", Daiichi Pure Chemicals
Co., LTD., Tokyo, Japan) and then pooled. The pooled GP is dialyzed
into 25 mM .beta.-glycerophosphate, 0.2 mM DTT, 0.3 mM EDTA, 200 mM
NaCl, pH 7.0 buffer (Buffer C) and stored on ice until use.
[0637] Prior to use of the GP enzyme, the enzyme is converted from
the inactive form as expressed in E. coli strain XL-1 Blue
(designated GPb) (Stragene Cloning Systems, La Jolla, Calif.), to
the active form (designated GPa) by the procedure described in
Section (A) Activation of GP below.
[0638] Purification of Glycogen Phosphorylase Expressed in Sf9
Cells
[0639] The Sf9 cells in pellets described above are resuspended in
25 mM .beta.-glycerophosphate (pH 7.0) with 0.2 mM DTT, 1 mM
MgCl.sub.2, plus the following protease inhibitors:
2 0.7 .mu.g/mL Pepstatin A 0.5 .mu.g/mL Leupeptin 0.2 mM
phenylmethylsulfonyl fluoride (PMSF), and 0.5 mM EDTA,
[0640] lysed by pretreatment with 3 .mu.g/mL DNAase followed by
sonication in batches for 3.times.1 minutes on ice using a Branson
Model 450 ultrasonic cell disrupter (Branson Sonic Power Co.,
Danbury Conn.). The Sf9 cell lysates are then cleared by
centrifugation at 35,000.times.g for one hour followed by
filtration through 0.45 micron filters. GP in the soluble fraction
of the lysates (estimated to be 1.5% of the total protein) is
purified by monitoring the enzyme activity (as described in GPa
Activity Assay section, below) from a series of chromatographic
steps detailed below.
[0641] Immobilized Metal Affinity Chromatography (IMAC)
[0642] Immobilized Metal Affinity Chromatography is performed as
described in the section above. The pooled, desalted GP is then
stored on ice until further processed.
[0643] Activation of GP
[0644] Before further chromatography, the fraction of inactive
enzyme as expressed in Sf9 cells (designated GPb) is converted to
the active form (designated GPa) by the following procedure
described in Section (A) Activation of GP below.
[0645] Anion Exchange Chromatography
[0646] Following activation of the IMAC purified GPb to GPa by
reaction with the immobilized phosphorylase kinase, the pooled GPa
fractions are dialyzed against 25 mM Tris-HCl, pH 7.5, containing
0.5 mM DTT, 0.2 mM EDTA, 1.0 mM phenylmethylsulfonyl fluoride
(PMSF), 1.0 .mu.g/mL leupeptin and 1.0 .mu.g/mL pepstatin A. The
sample is then loaded onto a MonoQ Anion Exchange Chromatography
column (Pharmacia Biotech. Inc., Piscataway, N.J.). The column is
washed with equilibration buffer until the A.sub.280 returns to
baseline. The sample is then eluted from the column with a linear
gradient of 0-0.25 M NaCl to remove the bound GP and other bound
proteins. GP-containing fractions elute between 0.1-0.2 M NaCl
range, as detected by monitoring the eluant for peak protein
absorbance at A.sub.280. The GP protein is then identified by
visualizing the M.sub.r approximately 97 kdal GP protein band by
sodium dodecyl sulfate polyacrylamide gel electrophoresis
(SDS-PAGE) followed by silver staining (2D-silver Stain II "Daiichi
Kit", Daiichi Pure Chemicals Co., LTD., Tokyo, Japan) and then
pooled. The pooled GP is dialyzed into 25 mM
N,N-bis[2-Hydroxyethyl]-2-aminoethanesulfonic acid, 1.0 mM DTT, 0.5
mM EDTA, 5 mM NaCl, pH 6.8 buffer and stored on ice until use.
[0647] Determination of GP Enzyme Activity
[0648] A) Activation of GP: Conversion of GPb to GPa
[0649] Prior to the determination of GP enzyme activity, the enzyme
is converted from the inactive form as expressed in E. coli strain
XL-1 Blue (designated GPb) (Stragene Cloning Systems, La Jolla,
Calif.), to the active form (designated GPa) by phosphorylation of
GP using phosphorylase kinase as follows. The fraction of inactive
enzyme as expressed in Sf9 cells (designated GPb) is also converted
to the active form (designated GPa) by the following procedure.
[0650] GP Reaction with Immobilized Phosphorylase Kinase
[0651] Phosphorylase kinase (Sigma Chemical Company, St. Louis,
Mo.) is immobilized on Affi-Gel 10 (BioRad Corp., Melvile, N.Y.) as
per the manufacturer's instructions. In brief, the phosphorylase
kinase enzyme (10 mg) is incubated with washed Affi-Gel beads (1
mL) in 2.5 mL of 100 mM HEPES and 80 mM CaCl.sub.2 at pH 7.4 for 4
hours at 4.degree. C. The Affi-Gel beads are then washed once with
the same buffer prior to blocking with 50 mM HEPES and 1 M glycine
methyl ester at pH 8.0 for one hour at room temperature. Blocking
buffer is removed and replaced with 50 mM HEPES (pH 7.4), 1 mM
.beta.-mercaptoethanol and 0.2% NaN.sub.3 for storage. Prior to use
to convert GPb to GPa, the Affi-Gel immobilized phosphorylase
kinase beads are equilibrated by washing in the buffer used to
perform the kinase reaction, consisting of 25 mM
.beta.-glycerophosphate, 0.3 mM DTT, and 0.3 mM EDTA at pH 7.8
(kinase assay buffer).
[0652] The partially purified, inactive GPb obtained from
5'-AMP-Sepharose chromatography above (from E. coli) or the mixture
of GPa and GPb obtained from IMAC above (from Sf9 cells) is diluted
1:10 with the kinase assay buffer then mixed with the
aforementioned phosphorylase kinase enzyme immobilized on the
Affi-Gel beads. NaATP is added to 5 mM and MgCl.sub.2 to 6 mM. The
resulting mixture is mixed gently at 25.degree. C. for 30 to 60
minutes. The sample is removed from the beads and the percent
activation of GPb by conversion to GPa is estimated by determining
GP enzyme activity in the presence and absence of 3.3 mM AMP. The
percentage of total GP enzyme activity due to GPa enzyme activity
(AMP-independent) is then calculated as follows: 1 % of total HLGPa
= HLGP activity - AMP HLGP activiy + AMP
[0653] Alternately, the conversion of GPb to GPa can be monitored
by isoelectric focusing, based on the shift in electrophoretic
mobility that is noted following conversion of GPb to GPa. GP
samples are analyzed by isoelectric focusing (IEF) utilizing the
Pharmacia PfastGel System (Pharmacia Biotech. Inc., Piscataway,
N.J.) using precast gels (pl range 4-6.5) and the manufacturer's
recommended method. The resolved GPa and GPb bands are then
visualized on the gels by silver staining (2D-silver Stain II
"Daiichi Kit", Daiichi Pure Chemicals Co., LTD., Tokyo, Japan).
Identification of GPa and GPb is made by comparison to E. coli
derived GPa and GPb standards that are run in parallel on the same
gels as the experimental samples.
[0654] B) GPa Activity Assay
[0655] The disease/condition treating/preventing activities
described herein of the glycogen phosphorylase inhibitor compounds
of this invention can be indirectly determined by assessing the
effect of the compounds of this invention on the activity of the
activated form of glycogen phosphorylase (GPa) by one of two
methods; glycogen phosphorylase a activity is measured in the
forward direction by monitoring the production of
glucose-1-phosphate from glycogen or by following the reverse
reaction, measuring glycogen synthesis from glucose-1-phosphate by
the release of inorganic phosphate. All reactions can be run in
triplicate in 96-well microtiter plates and the change in
absorbance due to formation of the reaction product is measured at
the wavelength specified below in a MCC/340 MKII Elisa Reader (Lab
Systems, Finland), connected to a Titertech Microplate Stacker (ICN
Biomedical Co, Huntsville, Ala.).
[0656] To measure the GPa enzyme activity in the forward direction,
the production of glucose-1-phosphate from glycogen is monitored by
the multienzyme coupled general method of Pesce et al. [Pesce, M.
A., Bodourian, S. H., Harris, R. C. and Nicholson, J. F. (1977)
Clinical Chemistry 23, 1711-1717] modified as follows: 1 to 100
.mu.g GPa, 10 units phosphoglucomutase and 15 units
glucose-6-phosphate dehydrogenase (Boehringer Mannheim
Biochemicals, Indianapolis, Ind.) are diluted to 1 mL in Buffer A
(described hereinafter). Buffer A is at pH 7.2 and contains 50 mM
HEPES, 100 mM KCl, 2.5 mM ethyleneglycoltetraacetic acid (EGTA),
2.5 mM MgCl.sub.2, 3.5 mM KH.sub.2PO.sub.4 and 0.5 mM
dithiothreitol. 20 .mu.l of this stock is added to 80 .mu.l of
Buffer A containing 0.47 mg/mL glycogen, 9.4 mM glucose, 0.63 mM of
the oxidized form of nicotinamide adenine dinucleotide phosphate
(NADP+). The compounds to be tested are added as 5 .mu.L of
solution in 14% dimethylsulfoxide (DMSO) prior to the addition of
the enzymes. The basal rate of GPa enzyme activity in the absence
of inhibitors is determined by adding 5 .mu.L of 14% DMSO and a
fully-inhibited rate of GPa enzyme activity is obtained by adding
20 .mu.L of 50 mM of the positive control test substance, caffeine.
The reaction is followed at room temperature by measuring the
conversion of oxidized NADP+ to reduced NADPH at 340 nm.
[0657] To measure the GPa enzyme activity in the reverse direction,
the conversion of glucose-1-phosphate into glycogen plus inorganic
phosphate is measured by the general method described by Engers et
al. [Engers, H. D., Shechosky, S. and Madsen, N. B. (1970) Can. J.
Biochem. 48, 746-754] modified as follows: 1 to 100 .mu.g GPa is
diluted to 1 mL in Buffer B (described hereinafter). Buffer B is at
pH 7.2 and contains 50 mM HEPES, 100 mM KCl, 2.5 mM EGTA, 2.5 mM
MgCl.sub.2 and 0.5 mM dithiothreitol. 20 .mu.L of this stock is
added to 80 .mu.L of Buffer B with 1.25 mg/mL glycogen, 9.4 mM
glucose, and 0.63 mM glucose-1-phosphate. The compounds to be
tested are added as 5 .mu.L of solution in 14% DMSO prior to the
addition of the enzyme. The basal rate of GPa enzyme activity in
the absence of added inhibitors is determined by adding 5 .mu.L of
14% DMSO and a fully-inhibited rate of GPa enzyme activity is
obtained by adding 20 .mu.L of 50 mM caffeine. This mixture is
incubated at room temperature for 1 hour and the inorganic
phosphate released from the glucose-1-phosphate is measured by the
general method of Lanzetta et al. [Lanzetta, P. A., Alvarez, L. J.,
Reinach, P. S. and Candia, O. A. (1979) Anal. Biochem. 100, 95-97]
modified as follows: 150 .mu.L of 10 mg/mL ammonium molybdate, 0.38
mg/mL malachite green in 1 N HCl is added to 100 .mu.L of the
enzyme mix. After a 20 minute incubation at room temperature, the
absorbance is measured at 620 nm.
[0658] The above assays carried out with a range of concentrations
of test compound allows the determination of an IC.sub.50 value
(concentration of test compound required for 50% inhibition) for
the in vitro inhibition of GPa enzyme activity by that test
compound.
[0659] Administration of the compounds of this invention can be via
any method which delivers a compound of this invention
preferentially to the desired tissue (e.g., liver and/or cardiac
tissues). These methods include oral routes, parenteral,
intraduodenal routes, etc. Generally, the compounds of the present
invention are administered in single (e.g., once daily) or multiple
doses or via constant infusion.
[0660] The compounds of this invention are useful, for example, in
reducing or minimizing damage effected directly to any tissue that
may be susceptible to either ischemia/reperfusion injury or injury
resulting from hypoxia (e.g., heart, brain, lung, kidney, liver,
gut, skeletal muscle, retina) as the result of an ischemic or
hypoxic event (e.g., myocardial infarction). The active compound is
therefore usefully employed prophylactically to prevent, i.e.
(prospectively or prophylactically) to blunt or stem, tissue damage
(e.g., myocardial tissue) in patients who are at risk for ischemia
or hypoxia (e.g., myocardial ischemia).
[0661] Generally, the compounds of this invention are administered
orally, or parenterally (e.g., intravenously, intramuscularly,
subcutaneously or intramedullary). Topical administration may also
be indicated, for example, where the patient is suffering from
gastrointestinal disorders or whenever the medication is best
applied to the surface of a tissue or organ as determined by the
attending physician.
[0662] The amount and timing of compounds administered will, of
course, be dependent on the subject being treated, on the severity
of the affliction, on the manner of administration and on the
judgement of the prescribing physician. Thus, because of patient to
patient variability, the dosages given below are a guideline and
the physician may titrate doses of the drug to achieve the
treatment that the physician considers appropriate for the patient.
In considering the degree of treatment desired, the physician must
balance a variety of factors such as age of the patient, presence
of preexisting disease, as well as presence of other diseases
(e.g., cardiovascular disease).
[0663] Thus, for example, in one mode of administration the
compounds of this invention may be administered just prior to
surgery (e.g., within twenty-four hours before surgery, for
example, cardiac surgery), during and/or subsequent to surgery
(e.g., within twenty-four hours after surgery) where there is risk
of ischemia (e.g., mycoardial ischemia). In another mode of
administration, the compounds of this invention are administered
with an initial loading dose (e.g., bolus injection or infusion)
prior to surgery followed by a constant infusion prior to, during
and post surgery. The compounds of this invention may also be
administered in a chronic daily mode.
[0664] An amount of a compound of this invention is used that is
effective for ischemic or hypoxic protection. A preferred dosage is
about 0.001 to about 100 mg/kg/day of a compound of this invention.
An especially preferred dosage is about 0.01 to about 50 mg/kg/day
of a compound of this invention.
[0665] The compounds of the present invention are generally
administered in the form of a pharmaceutical composition comprising
at least one of the compounds of this invention together with a
pharmaceutically acceptable vehicle or diluent. Thus, the compounds
of this invention can be administered individually or together in
any conventional oral, parenteral (e.g., intravenous, intramuscular
injection), rectal or transdermal dosage form.
[0666] For oral administration a pharmaceutical composition can
take the form of solutions, suspensions, tablets, pills, capsules,
powders, and the like. Tablets containing various excipients such
as sodium citrate, calcium carbonate and calcium phosphate are
employed along with various disintegrants such as starch and
preferably potato or tapioca starch and certain complex silicates,
together with binding agents such as polyvinylpyrrolidone, sucrose,
gelatin and acacia. Additionally, lubricating agents such as
magnesium stearate, sodium lauryl sulfate and talc are often very
useful for tabletting purposes. Solid compositions of a similar
type are also employed as fillers in soft and hard-filled gelatin
capsules; preferred materials in this connection also include
lactose or milk sugar as well as high molecular weight polyethylene
glycols. When aqueous suspensions and/or elixirs are desired for
oral administration, the compounds of this invention can be
combined with various sweetening agents, flavoring agents, coloring
agents, emulsifying agents and/or suspending agents, as well as
such diluents as water, ethanol, propylene glycol, glycerin and
various like combinations thereof.
[0667] For purposes of parenteral administration, solutions, for
example, in sesame or peanut oil or in aqueous propylene glycol can
be employed, as well as sterile aqueous solutions of the
corresponding water-soluble salts. Such aqueous solutions may be
suitably buffered, if necessary, and the liquid diluent first
rendered isotonic with sufficient saline or glucose. These aqueous
solutions are especially suitable for intravenous, intramuscular,
subcutaneous and intraperitoneal injection purposes. In this
connection, the sterile aqueous media employed are all readily
obtainable by standard techniques well-known to those skilled in
the art.
[0668] For purposes of transdermal (e.g., topical) administration,
dilute sterile, aqueous or partially aqueous solutions (usually in
about 0.1% to 5% concentration), otherwise similar to the above
parenteral solutions, are prepared.
[0669] Methods of preparing various pharmaceutical compositions
with a certain amount of active ingredient are known, or will be
apparent in light of this disclosure, to those skilled in this art.
For examples of methods of preparing pharmaceutical compositions,
see Reminqton's Pharmaceutical Sciences, Mack Publishing Company,
Easter, Pa., 15th Edition (1975).
[0670] Pharmaceutical compositions according to the invention may
contain, for example, 0.0001%-95% of the compound(s) of this
invention. In any event, the composition or formulation to be
administered will contain a quantity of a compound(s) according to
the invention in an amount effective to treat the disease/condition
of the subject being treated.
[0671] Advantageously, the present invention also provides kits for
use by a consumer having, or at risk of having, a disease or
condition resulting from, for example, ischemia or hypoxia, which
can be ameliorated by an A.sub.3 agonist. Such kits include a
suitable dosage form such as an injectable parenteral solution
particularly adapted for intravenous or intramuscular injection and
instructions describing the method of using such dosage form to
reduce the risk of tissue damage to the consumer. The instructions
would direct the consumer or medical personnel to administer the
parenteral solution according to administration modes known to
those skilled in the art. Such kits could advantageously be
packaged and sold in single or multiple parenteral kits units.
[0672] The two different compounds of the combination of this
invention can be co-administered simultaneously or sequentially in
any order, or as a single pharmaceutical composition comprising a
compound of Formula I and an aldose reductase inhibitor as
described above or a glycogen phosphorylase inhibitor as described
above or a sorbitol dehydrogenase inhibitor or a cardiovascular
agent.
[0673] Since the present invention has an aspect that relates to
the treatment of the disease/conditions described herein with a
combination of active ingredients which may be administered
separately, the invention also relates to combining separate
pharmaceutical compositions in kit form. The kit comprises two
separate pharmaceutical compositions: a compound of Formula I, a
prodrug thereof or a salt of such compound or prodrug and a second
compound as described above. The kit comprises a means for
containing the separate compositions such as a container, a divided
bottle or a divided foil packet. Typically, the kit comprises
directions for the administration of the separate components. The
kit form is particularly advantageous when the separate components
are preferably administered in different dosage forms (e.g., oral
and parenteral), are administered at different dosage intervals, or
when titration of the individual components of the combination is
desired by the prescribing physician.
[0674] An example of such a kit is a so-called blister pack.
Blister packs are well known in the packaging industry and are
being widely used for the packaging of pharmaceutical unit dosage
forms (tablets, capsules, and the like). Blister packs generally
consist of a sheet of relatively stiff material covered with a foil
of a preferably transparent plastic material. During the packaging
process recesses are formed in the plastic foil. The recesses have
the size and shape of the tablets or capsules to be packed. Next,
the tablets or capsules are placed in the recesses and the sheet of
relatively stiff material is sealed against the plastic foil at the
face of the foil which is opposite from the direction in which the
recesses were formed. As a result, the tablets or capsules are
sealed in the recesses between the plastic foil and the sheet.
Preferably the strength of the sheet is such that the tablets or
capsules can be removed from the blister pack by manually applying
pressure on the recesses whereby an opening is formed in the sheet
at the place of the recess. The tablet or capsule can then be
removed via said opening.
[0675] It may be desirable to provide a memory aid on the kit,
e.g., in the form of numbers next to the tablets or capsules
whereby the numbers correspond with the days of the regimen which
the tablets or capsules so specified should be ingested. Another
example of such a memory aid is a calendar printed on the card,
e.g., as follows "First Week, Monday, Tuesday, . . . etc . . .
Second Week, Monday, Tuesday, . . . " etc. Other variations of
memory aids will be readily apparent. A "daily dose" can be a
single tablet or capsule or several pills or capsules to be taken
on a given day. Also, a daily dose of Formula I compound can
consist of one tablet or capsule while a daily dose of the second
compound can consist of several tablets or capsules and vice versa.
The memory aid should reflect this.
[0676] In another specific embodiment of the invention, a dispenser
designed to dispense the daily doses one at a time in the order of
their intended use is provided. Preferably, the dispenser is
equipped with a memory aid, so as to further facilitate compliance
with the regimen. An example of such a memory aid is a mechanical
counter which indicates the number of daily doses that has been
dispensed. Another example of such a memory aid is a battery
powered microchip memory coupled with a liquid crystal readout, or
audible reminder signal which, for example, reads out the date that
the last daily dose has been taken and/or reminds one when the next
dose is to be taken.
[0677] The compounds of this invention generally will be
administered in a convenient formulation. The following formulation
examples are illustrative only and are not intended to limit the
scope of the present invention.
[0678] In the formulations which follow, "active ingredient" means
a compound(s) of this invention.
3 Formulation 1: Gelatin Capsules Hard gelatin capsules are
prepared using the following: Ingredient Quantity (mg/capsule)
Active ingredient 0.25-100 Starch, NF 0-650 Starch flowable powder
0-50 Silicone fluid 350 centistokes 0-15
[0679] A tablet formulation is prepared using the ingredients
below:
4 Formulation 2: Tablets Ingredient Quantity (mg/tablet) Active
ingredient 0.25-100 Cellulose, microcrystalline 200-650 Silicon
dioxide, fumed 10-650 Stearate acid 5-15
[0680] The components are blended and compressed to form
tablets.
[0681] Alternatively, tablets each containing 0.25-100 mg of active
ingredients are made up as follows:
5 Formulation 3: Tablets Ingredient Quantity (mg/tablet) Active
ingredient 0.25-100 Starch 45 Cellulose, microcrystalline 35
Polyvinylpyrrolidone (as 4 10% solution in water) Sodium
carboxymethyl cellulose 4.5 Magnesium stearate 0.5 Talc 1
[0682] The active ingredient, starch, and cellulose are passed
through a No. 45 mesh U.S. sieve and mixed thoroughly. The solution
of polyvinylpyrrolidone is mixed with the resultant powders which
are then passed through a No. 14 mesh U.S. sieve. The granules so
produced are dried at 50.degree.-60.degree. C. and passed through a
No. 18 mesh U.S. sieve. The sodium carboxymethyl starch, magnesium
stearate, and talc, previously passed through a No. 60 U.S. sieve,
are then added to the granules which, after mixing, are compressed
on a tablet machine to yield tablets.
[0683] Suspensions each containing 0.25-100 mg of active ingredient
per 5 ml dose are made as follows:
6 Formulation 4: Suspensions Quantity Ingredient (mg/5 ml) Active
ingredient 0.25-100 mg Sodium carboxymethyl cellulose 50 mg Syrup
1.25 mg Benzoic acid solution 0.10 mL Flavor q.v. Color q.v.
Purified Water to 5 mL
[0684] The active ingredient is passed through a No. 45 mesh U.S.
sieve and mixed with the sodium carboxymethyl cellulose and syrup
to form smooth paste. The benzoic acid solution, flavor, and color
are diluted with some of the water and 15 added, with stirring.
Sufficient water is then added to produce the required volume. An
aerosol solution is prepared containing the following
ingredients:
7 Formulation 5: Aerosol Ingredient Quantity (% by weight) Active
ingredient 0.25 Ethanol 25.75 Propellant 22 74.00
(Chlorodifluoromethane)
[0685] The active ingredient is mixed with ethanol and the mixture
added to a portion of the propellant 22, cooled to 30.degree. C.,
and transferred to a filling device. The required amount is then
fed to a stainless steel container and diluted with the remaining
propellant. The valve units are then fitted to the container.
[0686] Suppositories are prepared as follows:
8 Formulation 6: Suppositories Quantity Ingredient (mg/suppository)
Active ingredient 250 Saturated fatty acid glycerides 2,000
[0687] The active ingredient is passed through a No. 60 mesh U.S.
sieve and suspended in the saturated fatty acid glycerides
previously melted using the minimal necessary heat. The mixture is
then poured into a suppository mold of nominal 2 g capacity and
allowed to cool.
[0688] An intravenous formulation is prepared as follows:
9 Formulation 7: Intravenous Solution Ingredient Quantity Active
ingredient 25 mg-10,000 mg Isotonic saline 1,000 mL
[0689] The solution of the above ingredients is intravenously
administered to a patient.
[0690] The active ingredient above may also be a combination of
agents.
GENERAL EXPERIMENTAL PROCEDURES
[0691] NMR spectra were recorded on a Varian XL-300 (Varian Co.,
Palo Alto, Calif.), a Bruker AM-300 spectrometer (Bruker Co.,
Billerica, Mass.) or a Varian Unity 400 at about 23.degree. C. at
300 or 400 MHz for proton. Chemical shifts are expressed in parts
per million downfield from tetramethylsilane. The peak shapes are
denoted as follows: s, singlet; d, doublet; t, triplet, q, quartet;
m, multiplet; bs, =broad singlet. Resonances designated as
exchangeable did not appear in a separate NMR experiment where the
sample was shaken with several drops of D.sub.2O in the same
solvent. Atmospheric pressure chemical ionization mass spectra
(APCIMS) were obtained on a Fisons Platform II Spectrometer.
Chemical ionization mass spectra (CIMS) were obtained on a
Hewlett-Packard 5989 instrument (Hewlett-Packard Co., Palo Alto,
Calif.) (ammonia ionization, PBMS). Where the intensity of chlorine
or bromine-containing ions are described the expected intensity
ratio was observed (approximately 3:1 for
.sup.35Cl/.sup.37Cl-containing ions and 1:1 for
.sup.79Br/.sup.81Br-containing ions) and M is based on .sup.35Cl
and .sup.79Br. In some cases only representative .sup.1H NMR and
APCIMS peaks are given.
[0692] Column chromatography was performed with either Baker Silica
Gel (40 .mu.m) (J. T. Baker, Phillipsburg, N.J.) or Silica Gel 60
(EM Sciences, Gibbstown, N.J.) in glass columns or in Flash 40.TM.
or Flash 12.TM. (Biotage, Charlottesville, Va.) columns under
nitrogen pressure. Radial Chromatography was performed using a
Chromatron, (Harrison Research, Palo Alto, Calif.) Unless otherwise
specified, reagents were used as obtained from commercial sources.
Dimethylformamide, 2-propanol, tetrahydrofuran, and dichloromethane
used as reaction solvents were the anhydrous grade supplied by
Aldrich Chemical Company (Milwaukee, Wis.). Microanalyses were
performed by Schwarzkopf Microanalytical Laboratory, Woodside, N.Y.
The terms "concentrated" and "coevaporated" refer to removal of
solvent at water aspirator pressure on a rotary evaporator with a
bath temperature of less than 50.degree. C. The abbreviation "min"
and "h" stand for "minutes" and "hours" respectively and rt stands
for room temperature.
[0693] Reference to the hydrochloride salt in the Example names
below includes mono-or di-salts as appropriate in the particular
Example.
EXAMPLE 1
BOC Cleavage
[0694]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(3-methylisoxazol-5-ylmethoxy-
) benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic
acid methylamide.
(5-{6-[5-Chloro-2-(3-methylisoxazol-5-ylmethoxy)benzylamino]-
purin-9-yl}-4-hydroxy-2-methylcarbamoyltetrahydrofuran-3-yl)-carbamic
acid tert-butyl ester (1.0 mmol) was dissolved in anhydrous THF (10
mL). After adding H.sub.2O (10 mL) and then methanesulfonic acid
(1.5 mL, 15 mmol), the reaction was stirred for 6 h at 70.degree.
C. and then 15 h at room temperature. The organic solvent was
removed by rotary evaporation and the remaining aqueous solution
was neutralized to pH 7 with aqueous 1N NaOH solution. The title
compound then precipitated out and was recovered by filtration.
[0695] Mp 152.0-155.0.degree. C.; [.alpha.].sub.22=-30.5.degree.
(c=0.56, MeOH)
[0696] C.sub.23H.sub.25ClN.sub.8O.sub.5. MW 528.96. MS 529.1
(M+H).sup.+.
[0697] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.57 (s, 1H);
8.45 (quart, 1H, J=4.6 Hz); 8.35 (bs, 1H); 8.19 (s, 1H); 7.23 (dd,
1H, J=8.5 Hz, J=2.4Hz); 7.11 (d, 1H, J=8.5 Hz); 7.07 (bs, 1H); 6.48
(s, 1H); 5.99 (d, 1H, J=3.7 Hz); 5.87 (d, 1H, J=4.2 Hz); 5.29 (s,
2H); 4.62 (bs, 2H); 4.38-4.32 (mult, 1H); 4.08 (d, 1H. J=5.6 Hz);
3.58-3.51 (mult, 1H); 2.64 (d, 3H, J=4.6 Hz); 2.19 (s, 3H); 1.73
(bs, 2H).
EXAMPLE 2
Acetonide Cleavage
[0698]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(3,4,5,6-tetrahydroxytetrahyd-
ropyran-2-ylmethoxy)benzylamino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-ca-
rboxylic acid methyl amide.
[0699] To a solution of
3-amino-5-{-6-[5-chloro-2-(2,2,7,7-tetramethyltetr-
ahydro-bis[1,3]dioxolo[4,5-b;4',5'-d]pyran-5-ylmethoxy)
benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide (59 mg, 0.09 mmol) in chloroform (7 mL) was added
trifluoroacetic acid (0.7 mL). This reaction was stirred under
anhydrous conditions at room temperature for 2 h. After this time
period water was added (10 mL) and the reaction was stirred at room
temperature for 5 days. The solvent was removed with a rotary
evaporator and the resulting solid was then triturated with
Et.sub.2O to afford the title compound as a beige powder
[0700] (60 mg). Mp 212.0-218.0.degree. C.
C.sub.24H.sub.30ClN.sub.7O.sub.9- . MW 596.00. MS 596.1
(M+H).sup.+.
[0701] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.52-8.36 (mult,
5H); 8.35-8.24 (mult, 1H); 8.13 (s, 1H); 7.25-7.16 (mult, 1H);
7.10-6.90 (mult, 2H); 6.88-6.78 (mult, 1H); 6.21-6.16 (mult, 1H);
5.2-4.4 (mult, 2H); 5.02-4.88 (mult, 2H); 4.71-4.59 (mult, 2H);
4.53 (d, 1H, J=5.0 Hz); 4.30 (d, 1H, J=6.8 Hz); 4.26-4.20 (mult,
1H); 4.20-4.11 (mult, 1H); 4.10-3.99 (mult, 1H); 3.95-3.88 (mult,
1H); 3.84-3.76 (mult, 1H); 3.62-3.55 (mult, 1H); 3.34-3.25 (mult,
1H); 2.61 (d, 3H, J=4.4 Hz).
EXAMPLE 3
Reduction of Azide
[0702]
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzyloxy-5-chloro-benzylamino)-purin--
9-yl]-4-hydroxytetrahydrofuran-2-carboxylic acid methylamide.
[0703]
(2S,3S,4R,5R)-3-Azido-5-[6-(2-benzyloxy-5-chloro-benzylamino)-purin-
-9-yl-4-hydroxy tetrahydrofuran-2-carboxylic acid methyl amide (456
mg, 0.83 mmol) was dissolved in anhydrous THF (50 mL) and the
reaction cooled to 0.degree. C. After adding triphenylphosphine
(304 mg, 1.2 mmol), the reaction was stirred for 30 minutes at
0.degree. C. At the end of this time period, concentrated ammonium
hydroxide (0.4 mL) and water (0.5 mL) were added and the reaction
was allowed to slowly come to room temperature and stirred at room
temperature for 15 h. The solvent was then removed by rotary
evaporation and the product was preadsorbed onto silica gel and
purified by flash chromatography (SiO.sub.2, 5% then 18%
methanol/CH.sub.2Cl.sub.2) to afford the title compound as a
colorless solid.
[0704] Mp 114.2-115.2.degree. C.
[0705] [.alpha.].sub.22=-34-34.degree. (c=0.265, MeOH)
[0706] C.sub.25H.sub.26ClN.sub.7O.sub.4. MW 523.98. MS 524.1
(M+H).sup.+.
[0707] .sup.1H NMR (400 MHz, DMSO) .delta. 8.58 (s, 1H); 8.47
(quart, 1H, J=4.6 Hz); 8.35 (bs, 1H); 8.22 (s, 1H); 7.48-7.46
(mult, 2H); 7.38-7.33 (mult, 2H); 7.33-7.25 (mult, 1H); 7.25-7.20
(mult, 1H); 7.10-7.05 (mult, 2H); 6.01 (d, 1H, J=3.9 Hz); 5.95-5.85
(mult, 1H); 5.17 (s, 2H); 4.85-4.75 (mult, 2H); 4.38-4.34 (mult,
1H); 4.11 (d, 1H, J=5.8 Hz); 3.56 (t, 1H, J=5.8 Hz); 2.66 (d, 3H,
J=4.6 Hz); 1.9-1.7 (mult, 2H).
[0708] The following compounds, Examples 4-93 were prepared by
analogous procedures to the immediately preceding Example 3.
EXAMPLE 4
[0709]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2-morpholin-4-yl-ethoxy)-ben-
zylamino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide.
[0710] Mp 99.0-108.0.degree. C.
[0711] [.alpha.].sub.22=-29.64.degree. (c=0.280, MeOH)
[0712] C.sub.24H.sub.31ClN.sub.8O.sub.5. MW 547.02. MS 547.2
(M+H).sup.+.
[0713] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.57 (s, 1H);
8.45 (quart, 1H, J=4.6 Hz); 8.28 (bs, 1H); 8.21 (s, 1H); 7.22 (dd,
1H, J=8.7 Hz, J=2.4 Hz); 7.05 (bs, 1H); 7.01 (d, 1H, J=8.7 Hz);
6.02 (d, 1H, J=3.7 Hz), 6.05-5.80 (mult, 1H); 4.63 (bs, 2H);
4.40-4.30 (mult, 1H); 4.12 (t, 3H. J=5.5 Hz); 3.55-3.50 (mult, 5H);
2.70-2.60 (mult, 5H); 2.55-2.45 (mult, 4H); 2.25-1.95 (mult,
2H).
EXAMPLE 5
[0714]
(2S,3S,4R,5R)3-Amino-5-[6-(5-chloro-2-cyclobutylmethoxybenzylamino)-
-purin-9-yl]-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide.
[0715] Mp 107.0-117.0.degree. C.
[0716] [.alpha.].sub.21.5=-31.28.degree. (c=0.390, MeOH)
[0717] C.sub.23H.sub.28ClN.sub.7O.sub.4. MW 501.98; MS 501.9
(M+H).sup.+.
[0718] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.55 (s, 1H);
8.45-8.40 (mult, 1H); 8.25 (bs, 1H); 8.19 (s, 1H); 7.18 (dd, 1H,
J=8.5 Hz, J=2.7 Hz); 7.05-7.00 (mult, 1H); 6.95 (d, 1H, J=8.5 Hz);
5.99 (d, 1H, J=3.7 Hz); 5.90-5.80 (mult, 1H); 4.65-4.60 (mult, 2H);
4.35-4.30 (mult, 1H); 4.09 (d, 1H, J=5.8 Hz); 3.95 (d, 2H, J=6.2
Hz); 3.53 (t, 1H, J=5.8 Hz); 2.75-2.65 (mult, 1H); 2.63 (d, 3H,
J=4.6 Hz); 2.05-1.95 (mult, 2H); 1.90-1.80 (mult; 4H); 1.80-1.70
(mult, 2H).
EXAMPLE 6
[0719]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(3-methoxy-benzyloxy)-benzyla-
mino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0720] Mp 102.0-108.0.degree. C.
[0721] [.alpha.].sub.21.5=-28.890 (c=0.450, MeOH)
[0722] C.sub.26H.sub.28ClN.sub.7O.sub.5. MW 554.01. MS 553.8
(M+H).sup.+.
[0723] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.56 (s, 1H);
8.46 (quart, 1H, J=4.4 Hz); 8.35 (bs, 1H); 8.20 (s, 1H); 7.26 (t,
1H, J=8.1 Hz); 7.25 (dd, 1H, J=8.8 Hz, J=2.6 Hz); 7.10-7.05 (mult,
1H); 7.05-7.00 (mult, 3H); 6.84 (d, 1H, J=7.3 Hz); 5.99 (d, 1H,
J=3.7 Hz); 5.95-5.85 (mult, 1H); 5.13 (s, 2H); 4.75-4.65 (mult,
2H); 4.37 -4.30 (mult, 1H); 4.09 (d, 1H, J=5.6 Hz); 3.71 (s, 3H);
3.54 (t, 1H, J=4.9 Hz); 2.64 (d, 3H, J=4.4 Hz); 1.80-1.75 (mult,
2H).
EXAMPLE 7
[0724]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2,5-dimethoxy-benzyloxy)-ben-
zylamino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0725] Mp 112.0-115.0.degree. C.
[0726] [.alpha.].sub.21.5=-30.48.degree. (c=0.420, MeOH)
[0727] C.sub.27H.sub.30ClN.sub.7O.sub.6. MW 584.04. MS 583.8
(M+H).sup.+.
[0728] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.59 (s, 1H);
8.50 (quart, 1H, J=3.7 Hz); 8.37 (bs, 1H); 8.22 (s, 1H); 7.22 (dd,
1H, J=8.9 Hz, J=2.7 Hz); 7.10-7.05 (mult, 1H); 7.05-7.00 (mult,
2H); 6.97 (d, 1H, J=8.9 Hz); 6.86 (dd, 1H, J=8.9 Hz, J=2.7 Hz);
6.02 (d, 1H, J=3.9 Hz); 5.95-5.90 (mult, 1H); 5.10 (s, 2H);
4.75-4.65 (mult, 2H); 4.40-4.35 (mult, 1H); 4.12 (d, 1H, J=5.4 Hz);
3.78 (s, 3H); 3.67 (s, 3H); 3.60-3.55 (mult, 1H); 2.67 (d, 3H,
J=3.7 Hz); 1.85-1.75 (mult, 2H).
EXAMPLE 8
[0729]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(3-chloro-benzyloxy)-benzylam-
ino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0730] C.sub.25H.sub.25Cl.sub.2N.sub.7O.sub.4. MW 558.43. MS 557.8
(M+H).sup.+.
[0731] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.59 (s, 1H);
8.48 (quart, 1H, J=4.8 Hz); 8.40 (bs, 1H); 8.24 (s, 1H); 7.55 (s,
1H); 7.50-7.35 (mult, 3H); 7.24 (dd, 1H, J=8.7 Hz, J=2.7 Hz);
7.17-7.12 (bs, 1H); 7.05 (d, 1H, J=8.7 Hz); 6.02 (d, 1H, J=3.9 Hz);
5.95-5.85 (mult, 1H); 5.21 (s, 2H); 4.75-4.65 (mult, 2H); 4.40-4.35
(mult, 1H); 4.12 (d, 1H, J=5.8 Hz); 3.57 (t, 1H, J=5.8 Hz); 2.67
(d, 3H, J=4.8 Hz); 1.85-1.75 (mult, 2H).
EXAMPLE 9
[0732]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(4-chloro-benzyloxy)-benzylam-
ino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0733] Mp 88.0-92.0.degree. C.
[0734] [.alpha.].sub.24=-19.630.degree. (c=0.275, DMSO)
[0735] C.sub.25H.sub.25Cl.sub.2N.sub.7O.sub.4. MW 558.43. MS 558.1
(M+H).sup.+.
[0736] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 8.61 (s, 1H);
8.49 (quart, 1H, J=4.4 Hz); 8.39 (bs, 1H); 8.24 (s, 1H); 7.53 (d,
2H, J=8.9 Hz); 7.45 (d, 2H J=8.9 Hz); 7.23 (dd, 1H, J=8.7 Hz, J=3.6
Hz); 7.09 (bs, 1H); 7.02 (d, 1H, J=8.7 Hz); 6.04 (d, 1H, J=63 Hz);
5.95-5.87 (mult, 1H); 5.19 (s, 2H); 4.75-4.68 (mult, 2H); 4.41-4.36
(mult, 1H); 4.13 (d, 1H, J=5.1 Hz); 3.58 (t, 1H, J=5.1 Hz); 2.68
(d, 3H, J=4.4 Hz), 1.91 to 1.77 (mult, 2H).
EXAMPLE 10
[0737]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2-chloro-benzyloxy)-benzylam-
ino]-purin-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0738] Mp 88.0-92.0.degree. C.
[0739] [.alpha.].sub.24=-16.670.degree. (c=0.36, DMSO)
[0740] C.sub.25H.sub.25Cl.sub.2N.sub.7O.sub.4. MW 558.43. MS 558.1
(M+H).sup.+.
[0741] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 8.60 (s, 1H);
8.49 (quart, 1H, J=4.6 Hz); 8.36 (bs, 1H); 8.23 (s, 1H); 7.70-7.65
(mult, 1H); 7.55-7.50 (mult, 1H); 7.42-7.36 (mult, 2H); 7.27 (dd,
1H, J=8.5 Hz, J=2.6 Hz); 7.17-7.09 (mult, 2H); 6.03 (d, 1H, J=4.4
Hz); 5.95-5.88 (mult, 1H); 5.24 (s, 2H); 4.79-4.67 (mult, 2H);
4.40-4.36 (mult, 1H); 4.15 (d, 1H, J=6.3 Hz); 3.58 (t, 1H, J=6.3
Hz); 2.67 (d, 3H, J=4.6 Hz); 1.87-1.77 (mult, 2H).
EXAMPLE 11
[0742]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(tetrahydrofuran-3-ylmethoxy)-
-benzylamino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic
acid methylamide
[0743] Mp 114.0-118.0.degree. C.
[0744] C.sub.23H.sub.28ClN.sub.7O.sub.5. MW 517.98. MS 518.0
(M+H).sup.+.
[0745] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.58 (s, 1H);
8.47 (d, 1H, J=4.4 Hz); 8.31 (bs, 1H); 8.22 (s, 1H); 7.21 (dd, 1H,
J=8.5 Hz, J=2.7 Hz); 7.06 (bs, 1H); 6.99 (d, 1H, J=8.7 Hz); 6.01
(d, 1H, J=3.9 Hz); 5.89 (bs, 1H); 4.64 (bs, 2H); 4.36 (bs, 1H);
4.11 (d, 1H, J=5.6 Hz); 4.00-3.87 (mult, 2H); 3.80-3.70 (mult, 2H);
3.67-3.60 (mult, 1H); 3.59-3.50 (mult, 2H); 2.66 (d, 3H, J=4.4 Hz);
2.60 (bs, 1H); 2.04-1.96 (mult, 1H); 1.78 (bs, 2H); 1.74-1.62
(mult, 1H).
EXAMPLE 12
[0746]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(4-methyl-benzyloxy)-benzylam-
ino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0747] Mp 82.0-86.0.degree. C.
[0748] C.sub.26H.sub.28ClN.sub.7O.sub.4. MW 538.01. MS 538.2
(M+H).sup.+.
[0749] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.58 (bs, 1H);
8.48 (quart, 1H, J=4.8 Hz); 8.36 (bs, 1H); 8.20 (s, 1H); 7.45 (d,
1H, J=7.3 Hz); 7.27-7.15 (mult, 4H); 7.14 (d, 1H, J=8.7 Hz); 7.09
(bs, 1H); 6.01 (d, 1H, J=3.7 Hz); 5.90 (bs, 1H); 5.15 (s, 2H); 4.68
(bs, 2H); 4.38 (bs, 1H); 4.11 (d, 1H, J=5.8 Hz); 3.56 (t, 1H, J=5.8
Hz); 2.66 (d, 3H, J=4.8 Hz); 2.34 (s, 3H); 1.82 (bs, 2H).
EXAMPLE 13
[0750]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2-methyl-benzyloxy)-benzylam-
ino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0751] Mp 76.0-80.0.degree. C.
[0752] C.sub.26H.sub.28ClN.sub.7O.sub.4. MW 538.01. MS 538.2
(M+H).sup.+.
[0753] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.59 (s, 1H);
8.48 (quart, 1H, J=4.6 Hz); 8.35 (bs, 1H); 8.22 (s, 1H); 7.35 (d,
2H, J=8.1 Hz); 7.21-7.15 (mult, 3H); 7.22-7.17 (mult, 2H); 6.01 (d,
1H, J=3.9 Hz); 5.88 (bs, 1H); 5.12 (s, 2H); 4.70 (bs, 2H); 4.37
(bs, 1H); 4.11 (d, 1H, J=5.8 Hz); 3.56 (t, 1H, J=5.8 Hz); 2.66 (d,
3H, J=4.6 Hz); 2.28 (s, 3H); 1.79 (bs, 2H).
EXAMPLE 14
[0754]
(2S,3S4R,5R)3-Amino-5-{6-[5-chloro-2-(3-methyl-benzyloxy)-benzylami-
no]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0755] Mp 92.0-97.0.degree. C.
[0756] C.sub.26H.sub.28ClN.sub.7O.sub.4. MW 538.01. MS 538.2
(M+H).sup.+.
[0757] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.56 (s, 1H);
8.45 (quart, 1H, J=4.6 Hz); 8.32 (bs, 1H); 8.19 (s, 1H); 7.25-7.21
(mult, 3H); 7.21-7.16 (mult, 1H); 7.11-7.00 (mult, 3H); 5.99 (d,
1H, J=3.9 Hz); 5.87 (bs, 1H); 5.10 (s, 2H); 4.67 (bs, 2H); 4.34
(bs, 1H); 4.09 (d, 1H, J=5.8 Hz); 3.54 (t, 1H, J=5.8 Hz); 2.63 (d,
3H, J=4.6 Hz); 2.25 (s, 3H); 1.84
EXAMPLE 15
[0758]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2-methoxybenzyloxy)benzylami-
no]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0759] Mp 102.0-123.0.degree. C.
[0760] [.alpha.].sub.21=-49.39.degree. (c=0.225, MeOH)
[0761] C.sub.26H.sub.28ClN.sub.7O.sub.5. MW 554.01. MS 554.1
(M+H).sup.+.
[0762] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.58 (s, 1H);
8.48 (quart, 1H, J=4.6 Hz); 8.34 (bs, 1H); 8.21 (s, 1H); 7.44 (dd,
1H, J=7.5Hz, J=1.5 Hz); 7.30 (td, 1H, J=7.5 Hz, J=1.5 Hz); 7.22
(dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.08-7.01 (mult, 3H); 6.94 (t, 1H,
J=7.5 Hz); 6.02 (d, 1H, J=3.9 Hz); 5.91 (bs, 1H); 5.12 (s, 2H);
4.68 (bs, 2H); 4.37 (t, 1H, J=4.1 Hz); 4.11 (d, 1H, J=5.8 Hz); 3.82
(s, 3H); 3.57 (t, 1H, J=5.8 Hz); 2.66 (d, 3H, J=4.6 Hz); 1.91 (bs,
2H).
EXAMPLE 16
[0763]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(furan-3-ylmethoxy)benzylamin-
o]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0764] Mp 93.0-97.0.degree. C.
[0765] C.sub.23H.sub.24ClN.sub.7O.sub.5. MW 513.94. MS 513.8
(M+H).sup.+.
[0766] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.59 (s, 1H):
8.48 (quart, 1H, J=4.6 Hz); 8.34 (bs, 1H); 8.21 (s, 1H); 7.80 (s,
1H); 7.66 (s, 1H); 7.22 (dd, 1H, J=8.7 Hz, J=2.5 Hz); 7.10 (d, 1H,
J=8.7 Hz); 7.06 (bs, 1H); 6.60 (s, 1H); 6.01 (d, 1H, J=3.7 Hz);
5.90 (bs, 1H); 5.03 (s, 2H); 4.64 (bs, 2H); 4.36 (bs, 1H); 3.56 (t,
1H, J=4.5 Hz); 3.14 (d, 1H, J=5.2 Hz); 2.66 (d, 3H, J=4.6 Hz); 1.77
(bs, 2H).
EXAMPLE 17
[0767]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(4-methoxy-benzyloxy)benzylam-
ino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0768] C.sub.26H.sub.28ClN.sub.7O.sub.5. MW 554.01. MS 553.8
(M+H).sup.+.
[0769] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.55 (s, 1H);
8.44 (quart, 1H, J=4.4 Hz); 8.32 (bs, 1H); 8.19 (s, 1H); 7.37 (d,
2H, J=8.7 Hz); 7.18 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.11-7.04 (mult,
2H); 6.89 (d, 2H, J=8.7 Hz); 6.00 (d, 1H, J=3.7 Hz); 6.95 (bs, 1H);
5.06 (s, 2H); 4.62-4.59 (mult, 2H); 4.37 (bs, 1H); 4.11 (d, 1H,
J=5.4 Hz); 3.70 (s, 3H); 3.60-3.56 (mult, 1H); 2.63 (d, 3H, J=4.4
Hz).
EXAMPLE 18
[0770] (2S,3S,
4R,5R)3-Amino-5-[6-(5-chloro-2-cyclopentylmethoxy-benzylami-
no)-purin-9-yl]-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0771] Mp 110.6-116.2.degree. C.
[0772] [.alpha.].sub.22=-25.88.degree. (c=0.255, MeOH)
[0773] C.sub.24H.sub.30ClN.sub.7O.sub.4. MW 516.00. MS 515.8
(M+H).sup.+.
[0774] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.58 (s, 1H);
8.47 (quart, 1H, J=4.2 Hz); 8.29 (bs, 1H); 8.21 (s, 1H); 7.19 (dd,
1H, J=8.5 Hz, J=2.3 Hz); 7.03 (bs, 1H); 6.97 (d, 1H, J=8.5 Hz);
6.01 (d, 1H, J=3.1 Hz); 5.91 (bs, 1H); 4.64 (bs, 2H); 4.37 (bs,
1H); 4.11 (d, 1H, J=5.6 Hz); 3.87 (d, 2H, J=6. 4 Hz); 3.60-3.55
(mult, 1H); 2.65 (d, 3H, J=4.2 Hz); 2.36-2.22 (mult, 1H); 2.22-1.90
(mult, 2H); 1.80-1.70 (mult, 2H); 1.62-1.44 (mult, 4H); 1.39-1.28
(mult, 2H).
EXAMPLE 19
[0775]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[3-(2-morpholin-4-yl-ethoxy)--
benzyloxy]benzylamino}-purin-9-yl)-4-hydroxytetrahydrofuran-2-carboxylic
acid methylamide
[0776] C.sub.31H.sub.37ClN.sub.8O.sub.6. MW 653.14. MS 653.0
(M+H).sup.+.
[0777] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.57 (s, 1H);
8.45 (quart, 1H, J=4.4 Hz); 8.37 (bs, 1H); 8.22 (s, 1H); 7.26 (t,
1H, J=8.5Hz); 7.21 (dd, 1H, J=8.5 Hz, J=2.5 Hz); 7.09 (bs, 1H);
7.06-7.00 (mult, 3H); 6.87 (d, 1H, J=8.5 Hz); 6.02 (d, 1H, J=3.7
Hz); 5.95 (bs, 1H); 5.14 (s, 2H); 4.70 (bs, 2H); 4.42-4.36 (mult,
1H); 4.14 (d, 1H, J=5.4 Hz); 4.06 (t, 2H, J=5.4 Hz); 3.62-3.57
(mult, 1H); 3.53 (mult, 4H); 2.67-2.61 (mult, 5H); 2.42-2.38 (mult,
4H).
EXAMPLE 20
[0778]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(tetrahydrofuran-3-ylmethoxy)-
-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide
[0779] Mp 132.0-161.0.degree. C.
[0780] [60 ].sub.21=-16.47.degree. (c=0.170, MeOH)
[0781] C.sub.23H.sub.28ClN.sub.7O.sub.5. MW 517.98. MS 518.1
(M+H).sup.+.
[0782] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.53 (s, 1H);
8.42 (quart, 1H, J=4.4 Hz); 8.29 (bs, 1H); 8.19 (s, 1H); (dd, 1H,
J=8.7 Hz, J=2.7 Hz); 7.04 (s, 1H); 6.97 (d, 1H, J=8.7 Hz); 6.11
(bs, 1H), 6.01 (d, 1H, J=3.9 Hz); 4.61 (bs, 2H); 4.44-4.40 (mult,
1H); 3.78-3.65 4.15 (d, 1H, J=5.0 Hz); 3.99-3.92 (mult, 1H);
3.91-3.85 (mult, 1H); 3.78-3.65 (mult, 2H); 3.64-3.58 (mult, 2H);
3.57-3.50 (mult, 1H); 2.95 (quart, 1H, J=7.3 Hz); 2.69-2.58 (mult,
5H); 2.02-1.93 (mult, 1H); 1.70-1.59 (mult, 1H).
EXAMPLE 21
[0783]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(tetrahydro-furan-3-ylmethoxy-
)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide
[0784] C.sub.23H.sub.28ClN.sub.7O.sub.5. MW 517.98. MS 518.0
(M+H).sup.+.
[0785] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.54 (s, 1H);
8.47-8.40 (mult, 1H); 8.29 (bs, 1H); 8.19 (s, 1H); 7.18 (dd, 1H,
J=8.5 Hz, J=2.3 Hz); 7.03 (bs, 1H); 6.97 (d, 1H, J=8.5 Hz); 5.99
(d, 1H, J=3.7 Hz); 5.97 (bs, 1H); 4.61 (bs, 2H); 4.40-4.35 (mult,
1H); 4.11 (d, 1H, J=5.4 Hz); 3.98-3.92 (mult, 1H); 3.91-3.83 (mult,
1H); 3.78-3.68 (mult, 2H); 3.62-3.48 (mult, 3H); 2.87-2.82 (mult,
1H); 2.65-2.57 (mult, 5H); 2.02-1.93 (mult, 1H); 1.69-1.59 (mult,
1H).
EXAMPLE 22
[0786]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(furan-2-ylmethoxy)benzylamin-
o]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0787] Mp 187.0-192.0.degree. C.
[0788] C.sub.23H.sub.24ClN.sub.7O.sub.5. MW 513.95. MS 514.1
(M+H).sup.+.
[0789] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.55 (s, 1H);
8.44 (quart, 1H, J=4.4 Hz); 8.27 (bs, 1H); 8.20 (s, 1H); 7.67 (d,
1H, J=1.9 Hz); 7.21 (dd, 1H, J=8.9 Hz, J=2.7 Hz); 7.15 (d, 1H,
J=8.9 Hz); 7.02 (bs, 1H); 6.57 (d, 1H, J=3.3 Hz); 6.44 (dd, 1H,
J=3.3 Hz, J=1.9 Hz); 5.99 (d, 1H, J=4.0 Hz); 5.86 (bs, 1H); 5.11
(s, 2H); 4.58 (bs, 2H); 4.39-4.32 (mult, 1H); 4.09 (d, 1H, J=5.8
Hz); 3.59-3.53 (mult, 1H); 2.63 (d, 3H, J=4.4 Hz); 1.85 (bs,
2H).
EXAMPLE 23
[0790]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2,2,7,7-tetramethyltetrahydr-
o-bis[1,3]dioxolo[4,5-b;4',5'-d]pyran-5-ylmethoxy)benzylamino]purin-9-yl}--
4-hydroxytetrahydrofuran-2-carboxylic acid methylamide
[0791] C.sub.30H.sub.38ClN.sub.7O.sub.9. MW 676.13. MS 676.1
(M+H).sup.+.
[0792] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.55 (s, 1H);
8.47 (quart, 1H, J=4.4 Hz); 8.26 (bs, 1H); 8.22 (s, 1H); 7.22 (dd,
1H, J=8.7 Hz, J=2.3 Hz); 7.06 (bs, 1H); 7.02 (d, 1H, J=8.7 Hz);
6.02 (d, 1H, J=3.9 Hz); 5.48 (d, 1H, J=4.8 Hz); 4.65 (bs, 2H); 4.60
(dd, 1H, J=7.9 Hz, J=2.3 Hz); 4.42-4.39 (mult, 1H); 4.39-4.37
(mult, 1H); 4.37-4.28 (mult, 2H); 4.20-4.13 (mult, 2H); 4.09-3.98
(mult, 2H); 3.62 (t, 1H, J=5.4 Hz); 2.65 (d, 3H, J=4.4 Hz); 1.37
(s, 3H); 1.34 (s, 3H); 1.25 (s, 6H).
EXAMPLE 24
[0793]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2,5-dimethylfuran-3-ylmethox-
y)benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic
acid methylamide
[0794] Mp 85.0-88.0.degree. C.
[0795] C.sub.25H.sub.28ClN.sub.7O.sub.5. MW 542.00. MS 542.1
(M+H).sup.+.
[0796] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.58 (s, 1H);
8.48 (quart, 1H, J=4.6 Hz); 8.29 (bs, 1H); 8.20 (s, 1H); 7.22 (dd,
1H, J=8.7 Hz, J=2.3 Hz); 7.07 (d, 1H, J=8.7 Hz); 7.05 (bs, 1H);
6.07 (s, 1H); 6.01 (d, 1H, J=3.7 Hz); 5.89 (bs, 1H); 4.88 (s, 2H);
4.60 (bs, 2H); 4.36 (bs, 1H); 4.10 (d, 1H, J=5.4 Hz); 3.55 (t, 1H,
J=5.0 Hz); 2.66 (d, 3H, J=4.6 Hz); 2.23 (s, 3H); 2.16 (s, 3H); 1.77
(bs, 2H).
EXAMPLE 25
[0797]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(pyridin-3-ylmethoxy)benzylam-
ino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0798] Mp 200.0-218.0.degree. C.
[0799] C.sub.24H.sub.25ClN.sub.8O.sub.4. MW 524.97. MS 525.0
(M+H).sup.+.
[0800] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.69 (s, 1H);
8.52 (quart, 1H, J=4.6 Hz); 8.45 (s, 1H); 8.45-8.39 (mult, 1H);
8.35 (d, 1H, J=5.0 Hz); 8.20 (s, 1H); 7.89 (d, 1H, J=7.7 Hz); 7.40
(dd, 1H, J=7.7 Hz, J=4.8 Hz); 7.24 (dd, 1H, J=8.5 Hz, J=2.3 Hz);
7.11 (d, 1H, J=8.5 Hz); 6.17 (d, 1H, J=4.4 Hz); 5.22 (s, 2H);
4.90-4.84 (mult, 1H); 4.76-4.63 (mult, 2H); 4.60-4.56 (mult, 1H);
4.55-4.51 (mult, 1H); 4.18-4.09 (mult, 2H); 2.61 (d, 3H, J=4.6
Hz).
EXAMPLE 26
[0801]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(5-dimethylaminomethylfuran-2-
-ylmethoxy)benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic
acid methylamide
[0802] Mp 78.0-81.0.degree. C.
[0803] C.sub.26H.sub.31ClN.sub.8O.sub.5. MW 571.04. MS 571.1
(M+H).sup.+.
[0804] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.35 (s, 1H); 8.25
(s, 1H); 7.24 (bs, 1H); 7.20 (dd, 1H, J=8.7 Hz, J=2.5 Hz); 7.10 (d,
1H, J=8.7 Hz); 6.43 (d, 1H, J=3.1 Hz); 6.26 (d, 1H, J=3.1 Hz); 6.06
(d, 1H, J=4.2 Hz); 5.08 (s, 2H); 4.73 (bs, 2H); 4.59 (t, 1H, J=4.8
Hz); 4.30 (d, 1H, J=5.6 Hz); 3.74 (t, 1H, J=5.4 Hz); 3.48 (s, 2H);
2.82 (s, 3H); 2.20 (s, 6H).
EXAMPLE 27
[0805]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(thiazol-2-ylmethoxy)-benzyla-
mino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide
[0806] Mp 208.0-209.0.degree. C.
[0807] [60 ].sub.21=-32.08.degree. (c=0.265, MeOH)
[0808] C.sub.22H.sub.23ClN.sub.8O.sub.4S. MW 531.00. MS 531.0
(M+H).sup.+.
[0809] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.57 (s, 1H);
8.45 (quart, 1H, J=4.2 Hz); 8.38 (bs, 1H); 8.20 (s, 1H); 7.82 (d,
1H, J=3.2 Hz); 7.75 (d, 1H, J=3.2 Hz); 7.23 (dd, 1H, J=8.7 Hz,
J=2.3 Hz); 7.13 (d, 1H, J=8.7 Hz); 7.07 (s, 1H); 5.99 (d, 1H, J=3.5
Hz); 5.90-5.84 (mult, 1H); 5.49 (s, 2H); 4.69 (bs, 2H); 4.34 (bs,
1H); 4.09 (d, 1H, J=5.6 Hz); 3.59-3.50 (mult, 1H); 2.64 (d, 3H,
J=4.2 Hz); 1.74 (bs, 2H).
EXAMPLE 28
[0810]
(2S,3S,4R,5R)3-Amino-5-{6-[2-(benzothiazol-2-ylmethoxy)-5-chloroben-
zylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0811] Mp 127.0-129.0.degree. C.
[0812] C.sub.26H.sub.25ClN.sub.8O.sub.4S. MW 581.06. MS 581.0
(M+H).sup.+.
[0813] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.58 (s, 1H);
8.51-8.39 (mult, 2H); 8.21 (s, 1H); 8.09 (d, 1H, J=8.1 Hz); 8.00
(d, 1H, J=8.1 Hz); 7.50 (t, 1H, J=8.1 Hz); 7.42 (t, 1H, J=8.1 Hz);
7.25 (dd, 1H, J=8.7 Hz, J=2.5 Hz); 7.16 (d, 1H, J=8.7 Hz); 7.09
(bs, 1H); 6.00 (d, 1H, J=3.7 Hz); 5.91-5.84 (mult, 1H); 5.65 (s,
2H); 4.78 (bs, 2H); 4.34 (bs, 1H); 4.09 (d, 1H, J=5.8 Hz);
3.59-3.50 (mult, 1H); 2.64 (d, 3H, J=4.8 Hz); 1.74 (bs, 2H).
EXAMPLE 29
[0814]
(2S,3S,4R,5R)3-Amino-5-{6-[2-(benzofuran-2-ylmethoxy)-5-chlorobenzy-
lamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0815] Mp 127.0-130.0.degree. C.
[0816] C.sub.27H.sub.26ClN.sub.7O.sub.5. MW 564.01. MS 564.0.
(M+H).sup.+.
[0817] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.57 (s, 1H);
8.46 (quart, 1H, J=4.4 Hz); 8.33 (bs, 1H); 8.16 (s, 1H); 7.61 (d,
1H, J=7.5 Hz); 7.56 (d, 1H, J=8.3 Hz); 7.32-7.24 (mult, 1H);
7.24-7.19 (mult, 3H); 7.08-7.01 (mult, 2H); 5.99 (d, 1H, J=3.5 Hz);
5.87 (d, 1H, J=4.8 Hz); 5.33 (s, 2H); 4.63 (mult, 2H); 4.38-4.30
(mult, 1H); 4.08 (d, 1H, J=5.6 Hz); 3.59-3.48 (mult, 1H); 2.63 (d,
3H, J=4.4 Hz); 1.73 (bs, 2H).
EXAMPLE 30
[0818]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(isothiazol-5-ylmethoxy)benzy-
lamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0819] Mp 180.0-183.0.degree. C.
[0820] C.sub.22H.sub.23ClN.sub.8O.sub.4S. MW 531.00. MS 531.1
(M+H).sup.+.
[0821] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.59 (s, 1H);
8.53 (d, 1H, J=1.5 Hz); 8.47 (quart, 1H, J=4.8 Hz); 8.39 (bs, 1H);
8.22 (s, 1H); 7.47 (d, 1H, J=1.5 Hz); 7.26 (dd, 1H, J=8.7 Hz, J=2.7
Hz); 7.15-7.08 (mult, 2H); 6.02 (d, 1H, J=3.7 Hz); 5.90 (bs, 1H,
J=5.59 (s, 2H); 4.69 (bs, 2H); 4.36 (bs, 1H); 4.11 (d, 1H, J=5.6
Hz); 3.56 (t, 1H, J=5.6 Hz); 2.66 (d, 3H, J=4.8 Hz); 1.79 (bs,
2H).
EXAMPLE 31
[0822] (2S,3S,
4R,5R)3-Amino-5-{6-[5-chloro-2-(thiophen-2-ylmethoxy)benzyl-
amino]purin-9-yl}4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0823] Mp 135.0-141.0.degree. C.
[0824] C.sub.23H.sub.24ClN.sub.7O.sub.4S. MW 530.01. MS 530.1
(M+H).sup.+.
[0825] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.58 (s, 1H);
8.47 (quart, 1H, J=4.6 Hz); 8.35 (bs, 1H); 8.21 (s, 1H); 7.55 (dd,
1H, J=5.0 Hz, J=1.2 Hz); 7.27-7.19 (mult, 2H); 7.15 (d, 1H, J=8.7
Hz); 7.09-7.00 (mult, 2H); 6.07-5.87 (mult, 1H); 6.02 (d, 1H, J=3.3
Hz); 5.36 (s, 2H); 4.64 (bs, 2H); 4.41 (bs, 1H); 4.14 (d, 1H, J=5.8
Hz); 3.66-3.56 (mult, 1H); 2.65 (d, 3H, J=4.6 Hz).
EXAMPLE 32
[0826]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(quinolin-2-ylmethoxy)benzyla-
mino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[0827] Mp 130.0-134.0.degree. C.
[0828] C.sub.28H.sub.27ClN.sub.8O.sub.4. MW 575.03. MS 575.1
(M+H).sup.+.
[0829] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.57(s, 1H);
8.50-8.36 (mult, 3H); 8.21 (s, 1H); 8.02-7.93 (mult, 2H); 7.78-7.67
(mult, 2H); 7.62-7.53 (mult, 1H); 7.18 (d, 1H, J=8.9 Hz); 7.13 (bs,
1H); 7.06 (d, 1H, J=8.9 Hz); 6.00 (d, 1H, J=3.7 Hz); 5.88 (d, 1H,
J=4.6 Hz); 5.42 (s, 2H); 4.78 (bs, 2H); 4.34 (bs, 1H); 4.09 (d, 1H,
J=5.6 Hz), 3.57-3.48 (mult, 1H); 2.64 (d, 3H, J=4.6 Hz); 1.73 (bs,
2H).
EXAMPLE 33
[0830] (2S,3S,
4R,5R)3-Amino-5-{6-[5-chloro-2-(4-methyl-[1,2,3]thiadiazol--
5-ylmethoxy)benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic
acid methylamide
[0831] Mp 107.0-110.0.degree. C.
[0832] C.sub.22H.sub.24ClN.sub.9O.sub.4S. MW 546.01. MS 546.1
(M+H).sup.+.
[0833] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.60 (s, 1H);
8.48 (bs, 1H); 8.39 (bs, 1H); 8.20 (s, 1H); 7.36-7.23 (mult, 1H);
7.18 (d, 1H, J=8.1 Hz); 7.13 (bs, 1H); 6.02 (b, 1H); 5.92 (bs, 1H);
5.60 (s, 2H); 4.66 (bs, 2H); 4.36 (bs, 1H); 4.12 (bs, 1H); 3.57
(bs, 1H); 2.68 (bs, 6H); 1.78 (bs, 2H).
EXAMPLE 34
[0834]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(naphthalen-1-ylmethoxy)
benzylamino]purin-9-yl}-4-hydroxytetrahydro furan-2-carboxylic acid
methylamide
[0835] Mp 115.0-119.0.degree. C.
[0836] C.sub.29H.sub.28ClN.sub.7O.sub.4. MW 574.04. MS 574.9
(M+H).sup.+.
[0837] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.58 (s, 1H);
8.53-8.45 (mult, 1H); 8.34 (bs, 1H); 8.19 (s, 1H); 8.16 (d, 1H,
J=7.9 Hz); 7.99-7.86 (mult, 2H); 7.72 (d, 1H, J=6.4 Hz); 7.63-7.45
(mult, 3H); 7.35-7.23 (mult, 2H); 7.09 (bs, 1H); 6.01 (bs, 1H);
5.90 (bs, 1H); 5.63 (s, 2H); 4.64 (bs, 2H); 4.36 (bs, 1H); 4.11 (d,
1H, J=5.0 Hz); 3.56 (bs, 1H); 2.66 (d, 3H, J=4.2 Hz); 1.77 (bs,
2H).
EXAMPLE 35
[0838]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(3,5-dimethylisoxazol-4-ylmet-
hoxy) benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic
acid methylamide
[0839] C.sub.24H.sub.27ClN.sub.8O.sub.5. MW 542.99. MS 543.2
(M+H).sup.+.
[0840] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.55 (bs, 1H);
8.44 (bs, 1H); 8.31 (bs, 1H); 8.16 (s, 1H); 7.25 (d, 1H, J=8.5 Hz);
7.11 (d, 1H, J=8.7 Hz); 7.08 (bs, 1H); 5.98 (bs, 1H); 5.83 (bs,
1H); 4.95 (s, 2H); 4.58 (bs, 2H); 4.37 (bs, 1H); 4.11 (d, 1H, J=4.9
Hz); 3.57 (bs, 1H); 2.64 (d, 3H, J=3.8 Hz); 2.39 (s, 3H); 2.22 (s,
3H), 1.15 (bs, 2H).
EXAMPLE 36
[0841]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2-oxo-2-piperidin-1-yl-ethox-
y)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide. C.sub.25H.sub.31ClN.sub.8O.sub.5 M.W. 567.00. MS
567 (M+H).sup.+.
[0842] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.34 (s, 1H); 8.29
(bs, 1H); 7.3 (bs, 1H); 7.18 (d, 1H, J=8.9 Hz); 6.89 (d, 1H, J=8.9
Hz); 6.07 (d, 1H, J=4.1 Hz); 4.91 (s, 2H); 4.82 (bs, 2H); 4.64 (t,
1H, J=4.3 Hz); 4.38 (d, 1H, J=5.6 Hz); 3.79 (t, 1H, J=5.4 Hz); 3.51
(m, 4H); 2.8 (s, 3H); 1.6 (m, 6H).
EXAMPLE 37
[0843]
(2S,3S,4R,5R)3-Amino-5-[6-(5-chloro-2-phenylcarbamoylmethoxy-benzyl-
amino)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide.
[0844] C.sub.26H.sub.27ClN.sub.8O.sub.5 M.W. 567.00. MS 567
(M+H).sup.+.
[0845] 1H NMR (400 MHz, D6 DMSO) .delta. 8.6 (bs, 1H); 8.5 (bd, 1H,
J=5.5 Hz); 8.4 (bs, 1H); 7.6 (d, 2H, J=8.2 Hz); 7.33 (t, 2H, J=8.2
Hz); 7.23 (d, 1H, J=8.8 Hz); 7.12 (bs, 1H); 7.03 (t, 1H, J=8.2 Hz);
6.96 (d, 1H, J=8.8 Hz); 6.0 (d, 1H, J=4.0 Hz); 5.9 (bs, 1H); 4.8
(s, 2H); 4.78 (bs, 2H); 4.39 (bs, 1H); 4.1 (d, 1H, J=5.5 Hz); 3.59
(m, 1H); 2.62 (d, 3H, J=4.0 Hz); 1.94 (bs, 2H).
EXAMPLE 38
[0846]
(2S,3S,4R,5R)3-Amino-5-[6-(5-chloro-2dimethylcarbamoylmethoxy-benzy-
lamino)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide.
[0847] C.sub.22H.sub.27ClN.sub.8O.sub.5 M.W. 518.96. MS 519
(M+H).sup.+.
[0848] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.31 (s, 1H); 8.26
(bs, 1H); 7.28 (bs, 1H); 7.17 (dd, 1H, J=8.9, 2.5 Hz); 6.83 (d, 1H,
J=8.9 Hz); 6.03 (d, 1H, J=4.0 Hz); 4.9 (s, 2H); 4.82 (bs, 2H); 4.6
(t, 1H, J=4.6 Hz); 4.26 (d, 1H, J=5.6 Hz); 3.7 (t, 1H, J=5.6 Hz);
3.02 (s, 3H); 2.9 (s, 3H); 2.8 (s, 3H).
EXAMPLE 39
[0849]
(2S,3S,4R,5R)3-Amino-5-{6-[2-(benzylcarbamoyl-methoxy)-5-chloro-ben-
zylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide.
[0850] C.sub.27H.sub.29ClN.sub.8O.sub.5 M.W. 581.04. MS 581
(M+H).sup.+.
[0851] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.3 (s, 1H); 8.2
(s, 1H); 7.3 d, 1H, J=2.5 Hz); 7.19 (dd, 1H, J=8.8, 2.6 Hz); 6.9
(d, 1H, J=8.8 Hz); 6.02 (d, 1H, J=4.1 Hz); 4.82 (bs, 2H); 4.64 (s,
2H); 4.58 (t, 1H, J=4.5 Hz); 4.4 (s, 2H); 4.3 (d, 1H, J=5.6 Hz);
3.78 (t, 1H, J=4.8 Hz); 2.78 (s, 3H).
EXAMPLE 40
[0852]
(2R,3R,4S,5S)(2-{[9-(4-Amino-3-hydroxy-5-methylcarbamoyl-tetrahydro-
-furan-2-yl)-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetic
acid. C.sub.20H.sub.22ClN.sub.7O.sub.6 M.W.
[0853] 491.90. MS 492 (M+H).sup.+.
EXAMPLE 41
[0854]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(4-methyl-piperazin-1-yl)--
2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-carbox-
ylic acid methylamide. C.sub.25H.sub.32ClN.sub.9O.sub.5 M.W.
574.04. MS 574 (M+H).sup.+.
[0855] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.34 (s, 1H); 8.29
(bs, 1H); 7.29 (bs, 1H); 7.19 (d, 1H, J=8.9 Hz); 6.92 (d, 1H, J=8.9
Hz); 6.06 (d, 1H, J=4.1 Hz); 4.91 (s, 2H); 4.82 (bs, 2H); 4.59 (t,
1H, J=5.3 Hz); 4.30 (d, 1H, J=5.6 Hz); 3.79(t, 1H, J=5.6 Hz); 3.6
(m, 4H); 2.80 (m, 3H); 2.4 (m, 4H); 222 (s, 3H).
EXAMPLE 42
[0856]
(2S,3S,4R,5R)3-Amino-5-[6-(5-chloro-2-propylcarbamoylmethoxy-benzyl-
amino)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide.
[0857] C.sub.23H.sub.29ClN.sub.8O.sub.5 M.W. 532.99. MS 533
(M+H).sup.+.
[0858] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.37 (s, 1H); 8.29
(s, 1H); 7.27 d, 1H, J=2.5 Hz); 7.2 (dd, 1H, J=8.8, 2.6 Hz); 6.95
(D, 1H, J=8.8 Hz); 6.07 (d, 1H, J=4.0 Hz); 4.82 (bs, 2H); 4.6 (m,
3H); 4.3 (d, 1H, J=5.6 Hz); 3.8 (t, 1H, J=5.6 Hz); 3.2 (t, 2H,
J=7.1 Hz); 2.8 (s, 3H); 1.5 (m, 1H); 0.83 (t, 3H, J=7.1 Hz).
EXAMPLE 43
[0859]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2-morpholin-4-yl-2-oxo-ethox-
y)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide. C.sub.24H.sub.29ClN.sub.8O.sub.6 M.W. 561.00. MS
561 (M+H).sup.+.
[0860] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.35 (s, 1H); 8.28
(bs, 1H); 7.3 (bs, 1H); 7.2 (d, 1H, J=8.9 Hz); 6.94 (d, 1H, J=8.9
Hz); 6.07 (d, 1H, J=4.1 Hz); 4.94 (s, 2H); 4.82 (bs, 2H); 4.63 (t,
1H, J=5.3 Hz); 4.35 (d, 1H, J=5.6 Hz); 3.92 (t, 1H, J=5.6 Hz); 3.65
(m, 4H); 3.58 (m, 4H); 2.8 (s, 3H).
Example 44
[0861]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2-oxo-2-pyrrolidin-1-yl-etho-
xy)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide. C.sub.24H.sub.29ClN.sub.8O.sub.5 M.W. 545.00. MS
545 (M+H).sup.+.
[0862] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.28 (s, 1H); 8.25
(s, 1H); 7.24 (bs, 1H); 7.18 (d, 1H, J=8.9 Hz); 6.89 (d, 1H, J=8.9
Hz); 6.07 (d, 1H, J=3.9 Hz); 4.82 (bs, 2H); 4.8 (s, 2H); 4.64 (t,
1H, J=5.3 Hz); 4.38 (d, 1H, J=5.6 Hz); 3.92 (t, 1H, J=5.6 Hz); 3.51
(t, 2H, J=6.7 Hz); 3.41 (t, 3H, J=6.7 Hz); 2.8 (s, 3H); 1.92 (m,
2H); 1.82 (m, 2H).
EXAMPLE 45
[0863]
(2S,3S,4R,5R)3-Amino-5-[6-(5-chloro-2-dipropylcarbamoylmethoxy-benz-
ylamino)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide.
[0864] C.sub.26H.sub.35ClN.sub.8O.sub.5 M.W. 575.07. MS 575
(M+H).sup.+.
[0865] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.32 (s, 1H); 8.27
(bs, 1H); 7.28 (bs, 1H); 7.2 (d, 1H, J=8.9 Hz); 6.86 (d, 1H, J=8.9
Hz); 6.04 (d, 1H, J=3.8 Hz); 4.92 (s, 2H); 4.82 (bs, 2H); 4.6 (t,
1H, J=5.5 Hz); 4.3 (d, 1 H, J=5.6 Hz); 3.8 (t, 1H, J=Hz); 3.3 (m,
4H); 2.8 (s, 3H); 1.65 (m, 1H); 1.48 (m, 1H); 0.97 (t, 3H, J=7.2
Hz); 0.88 (t, 3H, J=7.2 Hz).
EXAMPLE 46
[0866]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[(2-methoxy-ethylcarbamoyl)-m-
ethoxy]-benzylamino)-purin-9-yl)4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide. C.sub.23H.sub.29ClN.sub.8O.sub.6 M.W. 548.99. MS
549 (M+H).sup.+.
[0867] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.37 (s, 1H); 8.3
(s, 1H); 7.3 (d, 1H, J=2.5 Hz); 7.2 (dd, 1H, J=8.8, 2.5 Hz); 6.95
(d, 1H, J=8.8 Hz); 6.07 (d, 1H, J=4.0 Hz); 4.82 (bs, 2H); 4.6 (m,
3H); 4.3 (d, 1H, J=5.8 Hz); 3.8 (m, 1H); 3.41 (s, 3H); 3.3 (m, 4H);
2.8 (s, 3H).
EXAMPLE 47
[0868]
(2S,3S,4R,5R)3-Amino-5-[6-(5-chloro-2-methylcarbamoylmethoxy-benzyl-
amino)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide.
[0869] C.sub.21H.sub.25ClN.sub.8O.sub.5 M.W. 504.93. MS 505
(M+H).sup.+.
[0870] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.32 (s, 1H); 8.26
(s, 1H); 7.24 (d, 1H, J=2.5 Hz); 7.2 (dd, 1H, J=8.9, 2.5 Hz); 6.83
(d, 1H, J=8.9 Hz); 6.04 (d, 1H, J=3.9 Hz); 4.82 (bs, 2H); 4.6 (t,
1H, J=5.5 Hz); 4.58 (s, 2H); 4.3 (d, 1H, J=5.6 Hz); 3.8 (t, 1H,
J=5.6 Hz); 2.8 (s, 6H).
EXAMPLE 48
[0871]
(2S,3S,4R,5R)3-Amino-5-[6-(5-chloro-2-cyclohexylcarbamoylmethoxy-be-
nzylamino)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide. C.sub.26H.sub.33ClN.sub.8O.sub.5 M.W. 573.05. MS 573
(M+H).sup.+. .sup.1H NMR (400 MHz, D6 DMSO) d 8.6 (s, 1H); 8.43 (m,
1H); 8.4 (bs, 1H); 8.2 (s, 1H); 7.8 (bd, 1H, J=9 Hz); 7.2 (d, 1H,
J=8.8 Hz); 7.1 (bs, 1H); 6.82 (d, 1H, J=8.8 Hz); 6.0 (d, 1H, J=4.0
Hz); 5.95 (bs, 1H); 4.7 (bs, 2H); 4.52 (s, 2H); 4.4 (m, 1H); 4.1
(d, 1H, J=5.7 Hz); 3.6 (m, 2H); 2.62 (d, 3H, J=4.2 Hz); 1.65 (m,
4H); 1.5 (m, 1H); 1.2 (m, 5H).
EXAMPLE 49
[0872]
(2R,3R,4S,5S)4-[(2-{[9-(4-Amino-3-hydroxy-5-methylcarbamoyl-tetrahy-
dro-furan-2-yl)-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetyl]-pipe-
razine-1-carboxylic acid ethyl ester.
C.sub.27H.sub.34ClN.sub.9O.sub.7 M.W. 632.07. MS 632 (M+H).sup.+.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33 (s, 1H); 8.28 (s,
1H); 7.3 (bs, 1H); 7.2 (d, 1H, J=8.7 Hz); 6.93 (d, 1H, J=8.7 Hz);
6.07 (d, 1H, J=3.9 Hz); 4.93 (s, 2H); 4.82 (bs, 2H); 4.6 (t, 1H,
J=5.5 Hz); 4.3 (d, 1H, J 5.6 Hz); 4.1 (q, 2H, J=7.1 Hz); 3.79 (t,
1H, J=5.5 Hz); 3.6 (m, 4H); 3.42 (m, 4H); 2.8 (s, 3H); 1.22 (t, 3H,
J=7.1 Hz).
EXAMPLE 50
[0873]
(2S,3S,4R,5R)3-Amino-5-{6-[2-(2-azetidin-1-yl-2-oxo-ethoxy)-5-chlor-
o-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide. C.sub.23H.sub.27ClN.sub.8O.sub.5 M.W. 530.97. MS
531 (M+H).sup.+.
[0874] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33 (s, 1H); 8.27
(s, 1H); 7.3 (s, 1H); 7.2 (d, 1H, J=8.7 Hz); 6.85 (d, 1H, J=8.7
Hz); 6.07 (d, 1H, J=4.1 Hz); 4.82 (bs, 2H); 4.63 (s, 2H); 4.61 (t,
1H, J=5.3 Hz); 4.37 (m, 3H); 4.02 (m, 2H); 3.82 (t, 1H, J=5.6 Hz);
2.8 (s, 3H); 2.3 (m, 2H).
EXAMPLE 51
[0875]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[(2-morpholin-4-yl-ethylcarba-
moyl-methoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-carbox-
ylic acid methylamide. C.sub.26H.sub.34ClN.sub.9O.sub.6 M.W.
604.07. MS 604 (M+H).sup.+.
[0876] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.36 (s, 1H); 8.29
(s, 1H); 7.3 (s, 1H); 7.2 (d, 1H, J=8.7 Hz); 6.93 (d, 1H, J=8.7
Hz); 6.08 (d, 1H, J=3.7 Hz); 4.84 (bs, 2H); 4.63 (t, 1H, J=5.5 Hz);
4.6 (s, 2H); 4.38 (d, 1H, J=5.6 Hz); 3.9 (t, 1H, J=5.3 Hz); 3.55
(m, 4H); 3.4 (t, 2H, J=6.4 Hz); 2.8 (s, 3H); 2.42 (t, 2H, J=6.4
Hz); 2.4 (m, 4H).
EXAMPLE 52
[0877]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-oxo-2-(4-phenyl-piperazin--
1-yl)-ethoxyl]-benzylamino}-purin-9-yl)4-hydroxy-tetrahydro-furan-2-carbox-
ylic acid methylamide. C.sub.30H.sub.34ClN.sub.9O.sub.5 M.W.
636.12. MS 636 (M+H).sup.+.
[0878] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.32 (s, 1H); 8.26
(s, 1H); 7.3 (s, 1H); 7.2 (m, 3H); 6.9 (m, 4H); 6.07 (d, 1H, J=3.9
Hz); 4.92 (s, 2H); 4.82 (bs, 2H); 4.59 (t, 1H, J=5.3 Hz); 4.3 (d,
1H, J=5.6 Hz); 3.72(m, 5H); 3.11 (m, 4H); 2.8 (s, 3H).
EXAMPLE 53
[0879]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(4-cyclohexyl-piperazin-1--
yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)4-hydroxy-tetrahydro-furan-2-car-
boxylic acid methylamide. C.sub.30H.sub.40ClN.sub.9O.sub.5 M.W.
642.16. MS 642 (M+H).sup.+.
[0880] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33 (s, 1H); 8.29
(s, 1H); 7.3 (s, 1H); 7.2 (dd, 1H, J=8.8, 2.6 Hz); 6.93 (d, 1H,
J=8.8 Hz); 6.08 (d, 1H, J=3.8 Hz); 4.93 (s, 2H); 4.82 (bs, 2H);
4.63 (t, 1H, J=5.5 Hz); 4.38 (d, 1H, J=5.6 Hz); 3.83(m, 1H); 3.6
(m, 4H); 2.8 (s, 3H); 2.6 (m, 4H); 2.32 (m, 1H); 1.8-1.2 (m,
10H).
EXAMPLE 54
[0881]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(4-ethyl-piperazin-1-yl)-2-
-oxo-ethoxyl]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-carbox-
ylic acid methylamide. C.sub.26H.sub.34ClN.sub.9O.sub.5 M.W.
588.07. MS 588 (M+H).sup.+.
[0882] .sup.1H NMR (400 MHz, CD.sub.3OD) 8.32 (s, 1H); 8.28 (s,
1H); 7.3 (bs, 1H); 7.2 (d, 1H, J=8.7 Hz); 6.93 (d, 1H, J=8.7 Hz);
6.07 (d, 1H, J=3.9 Hz); 4.93 (s, 2H); 4.82 (s, 2H); 4.64 (t, 1H,
J=5.3 Hz); 4.38 (d, 1H, J=6.4 Hz); 3.9 (t, 1H, J=5.5 Hz); 3.6 (bs,
4H); 2.8 (s, 3H); 2.45 (m, 6H); 1.05 (t, 3H, J=7.3 Hz).
EXAMPLE 55
[0883]
(2S,3S,4R,5R)3-Amino-5-[6-(5-chloro-2-cyclopropylcarbamoylmethoxy-b-
enzylamino)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide. C.sub.23H.sub.27ClN.sub.8O.sub.5 M.W. 530.98. MS
531 (M+H).sup.+. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.37 (s,
1H); 8.31 (s, 1H); 7.3 (bs, 1H); 7.2 (dd, 1H, J=8.6, 2.6 Hz); 6.85
(d, 1H, J=8.6 Hz); 6.07 (d, 1H, J=3.9 Hz); 4.82 (bs, 2H); 4.58 (m,
3H); 4.3 (d, 1H, J=5.6 Hz); 3.75 (t, 1H, J=5.6 Hz); 2.8 (s, 3H);
2.63 (m, 1H); 0.7 (m, 2H); 0.5 (m, 2H).
EXAMPLE 56
[0884]
(2S,3S,4R,5R)3-Amino-5-[6-(2-carbamoylmethoxy-5-chloro-benzylamino)-
-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide. C.sub.20H.sub.23ClN.sub.8O.sub.5 M.W. 490.91. MS 491
(M+H).sup.+.
[0885] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.37 (s, 1H); 8.31
(s, 1H); 7.36 (d, 1H, 2.65 Hz); 7.25 (dd, 1H, J=8.6, 2.6 Hz); 6.96
(d, 1H, J=8.6 Hz); 6.09 (d, 1H, J=4.28 Hz); 4.88 (s, 2H); 4.62 (m,
3H); 4.33 (d, 1H, J=5.6 Hz); 3.77 (t, 1H, J=5.6 Hz); 2.84 (s,
3H).
EXAMPLE 57
[0886]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(4-cyclopropyl-piperazin-1-
-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-c-
arboxylic acid methylamide. C.sub.27H.sub.34ClN.sub.9O.sub.5 M.W.
600.08. MS 600 (M+H).sup.+.
[0887] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.32 (s, 1H); 8.28
(s, 1H); 7.3 (bs, 1H); 7.2 (dd, 1H, J=8.8, 2.8 Hz); 6.93 (d, 1H,
J=8.8 Hz); 6.07 (d, 1H, J=4.06 Hz); 4.89 (s, 2H); 4.83 (bs, 2H);
4.64 (t, 1H, J=4.9 Hz); 4.35 (d,1H, J=5.8 Hz); 3.86 (t, 1H, J=5.7
Hz); 3.53 (m, 4H); 2.80 (s, 3H); 2.60 (m, 4H); 1.63 (m, 1H); 0.47
(m, 2H); 0.42 (m, 2H).
EXAMPLE 58
[0888]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(4-isopropyl-piperazin-1-y-
l)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)4-hydroxy-tetrahydro-furan-2-carb-
oxylic acid methylamide. C.sub.27H.sub.36ClN.sub.9O.sub.5 M.W.
602.1. MS 602 (M+H).sup.+.
[0889] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.28 (s, 1H); 8.27
(s, 1H); 7.29 (bs, 1H); 7.2 (dd, 1H, J=8.8, 2.6 Hz); 6.9 (d, 1H,
J=8.8 Hz); 6.11 (d, 1H, J=3.2 Hz); 4.93 (s, 2H); 4.9 (m, 1H); 4.82
(s, 2H); 4.53 (d, 1H, J=6.2 Hz); 4.28 (t, 1H, J=5.5 Hz); 3.74 (bs,
4H); 3.04 (m, 1H); 2.84 (m, 4H); 2.75 (s, 3H); 1.66 (d, 6H, J=6.4
Hz).
EXAMPLE 59
[0890]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-oxo-2-(4-propyl-piperazin--
1-yl)-ethoxyl]-benzylamino}-purin-9-yl)4-hydroxy-tetrahydro-furan-2-carbox-
ylic acid methylamide. C.sub.27H.sub.36ClN.sub.9O.sub.5 M.W. 602.1.
MS 602 (M+H).sup.+.
[0891] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.26 (s, 2H); 7.28
(bs, 1H); 7.2 (dd, 1H, J=8.4, 2.2 Hz); 6.93 (d, 1H, J=8.4 Hz); 6.1
(d, 1H, J=3.2 Hz); 4.92 (s, 2H); 4.9 (m, 1H); 4.82 (s, 2H); 4.54
(d, 1H, J=6.4 Hz); 4.33 (t, 1H, J=6.19 Hz); 3.66 (bs, 4H); 2.74 (s,
3H); 2.66 (m, 4H); 2.49 (t, 2H, J=7.9 Hz); 1.57 (m, 2H); 0.93 (t,
3H, J=7.2 Hz).
EXAMPLE 60
[0892]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(4-cyclopentyl-piperazin-1-
-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-c-
arboxylic acid methylamide. C.sub.29H.sub.38ClN.sub.9O.sub.5 M.W.
628.14. MS 628 (M+H).sup.+.
[0893] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.31 (s, 1H); 8.28
(s, 1H); 7.3 (s,1H); 7.2 (dd, 1H, J=8.6, 2.6 Hz); 6.93 (d, 1H,
J=8.6 Hz); 6.08 (d, 1H, J=3.8 Hz); 4.89 (s, 2H); 4.82 (s, 2H); 4.70
(t, 1H, J=4.8 Hz); 4.4 (d, 1H, J=5.9 Hz); 3.98(t, 1H, J=5.6 Hz);
3.6 (m, 4H); 2.78 (s, 3H); 2.55 (m, 4H); 1.9-1.2 (m, 9H).
EXAMPLE 61
[0894]
(2S,3S,4R,5R)3-Amino-5-(6-{2-[2-(4-benzyl-piperazin-1-yl)-2-oxo-eth-
oxy]-5-chloro-benzylamino}-purin-9-yl)4-hydroxy-tetrahydro-furan-2-carboxy-
lic acid methylamide. C.sub.31H.sub.36ClN.sub.9O.sub.5 M.W. 650.1.
MS 650 (M+H).sup.+.
[0895] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.30 (s, 1H); 8.27
(s, 1H); 7.27 (m, 7H); 6.90 (d, 1H, J=8.9 Hz); 6.06 (d, 1H, J=4.1
Hz); 4.85 (s, 2H); 4.81 (s, 2H); 4.61 (t, 1H, J=4.5 Hz); 4.32 (d,
1H, J=5.7 Hz); 3.79(t, 1H, J=5.6 Hz); 3.56 (m, 4H); 3.5(s, 2H);
2.79 (s, 3H); 2.43 (m, 4H).
EXAMPLE 62
[0896]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-oxo-2-(3-oxo-piperazin-1-y-
l)-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-carboxyli-
c acid methylamide. C.sub.24H.sub.28ClN.sub.9O.sub.6 M.W. 574.00.
MS 574 (M+H).sup.+.
[0897] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.32 (s, 1H); 8.27
(s, 1H); 7.30 (s, 1H); 7.2 (m, 1H); 6.94 (d, 1H, J=8.7 Hz); 6.06
(d, 1H, J=3.9 Hz); 4.93 (s, 2H); 4.82 (bs, 2H); 4.62 (t, 1H, J=4.5
Hz); 4.33 (d, 1H, J=5.7 Hz); 4.13(s, 1H); 3.80 (m 1H); 3.8 (t, 1H,
J=4.5 Hz); 3.76 (bs, 2H); 3.38 (m, 1H); 3.30 (m, 1H); 2.80 (s,
3H).
EXAMPLE 63
[0898]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2-oxo-2-piperazin-1-yl-ethox-
y)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide. C.sub.24H.sub.30ClN.sub.9O.sub.5 M.W. 560.01. MS
560 (M+H).sup.+.
[0899] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.34 (s, 1H); 8.28
(s, 1H); 7.29 (s, 1H); 7.19 (dd, 1H, J=8.6, 2.6 Hz); 6.92 (d, 1H,
J=8.9 Hz); 6.06 (d, 1H, J=4.3 Hz); 4.89 (s, 2H); 4.82 (s, 2H); 4.59
(dd, 1H, J=5.0, 4.5 Hz); 4.30 (d, 1H, J=5.6 Hz); 3.74(d, 1H, J=5.6
Hz); 3.54 (m, 4H); 2.80 (m, 7H).
EXAMPLE 64
[0900]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(4-ethyl-3-oxo-piperazin-1-
-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-c-
arboxylic acid methylamide. C.sub.26H.sub.32ClN.sub.9O.sub.6 M.W.
602.1. MS 602 (M+H).sup.+.
[0901] .sup.1H NMR (400 MHz. CD.sub.3OD) .delta. 8.33 (s, 1H); 8.28
(s, 1H); 7.30 (m, 1H); 7.2 (t, 1H, J=6.2 Hz); 6.94 (dd, 1H, J=8.7,
3.9 Hz); 6.06 (d, 1H, J=4.1 Hz); 4.92 (s, 2H); 4.83 (m, 2H); 4.61
(t, 1H, J=4.2 Hz); 4.32 (d, 1H, J=5.7 Hz); 4.22 (s, 1H); 4.13 (s,
1H); 3.8 (m, 3H); 3.46 (m, 4H); 2.81 (s, 3H); 1.12 (q, 3H,
J=7.3).
EXAMPLE 65
[0902]
(2S,3S,4R,5R)3-Amino-5-[6-(5-chloro-2-{2-[4-(2-chloro-phenyl)-piper-
azin-1-yl]-2-oxo-ethoxy}-benzylamino)-purin-9-yl]-4-hydroxy-tetrahydro-fur-
an-2-carboxylic acid methylamide. C.sub.30H.sub.33ClN.sub.9O.sub.5
M.W. 670.55. MS 670 (M+H).sup.+. .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta.8.29 (s, 1H); 8.23 (s, 1H); 7.33 (m, 2H); 7.2 (m, 2H); 6.97
(m, 3H); 6.03 (d, 1H, J=4.2 Hz); 4.92 (s, 2H); 4.83 (s, 2H); 4.56
(t, 1H, J=4.6 Hz); 4.28 (d, 1H, J=5.6 Hz); 3.73(m, 5H); 2.98 (m,
4H); 2.77 (s, 3H).
EXAMPLE 66
[0903]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(phenethylcarbamoyl-methoxy)--
benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide. C.sub.28H.sub.31ClN.sub.8O.sub.5 M.W. 595.06. MS
595 (M+H).sup.+.
[0904] .sup.1H NMR (400 MHz. CD.sub.3OD) .delta. 8.37 (s,1H); 8.27
(s, 1H); 7.3 (d, 1H, J=2.5 Hz); 7.2-7.02 (m, 6H); 6.83 (d, 1H,
J=8.8 Hz); 6.06 (d, 1H, J=4.3 Hz); 4.76 (s, 2H); 4.57 (m, 3H); 4.3
(d, 1H, J=5.7 Hz); 3.74 (t, 1H, 5.6 Hz); 3.5 (t, 4H, 7 Hz); 2.80
(s, 3H); 2.76 (t, 1H, J=7 Hz).
EXAMPLE 67
[0905]
(2S,3S,4R,5R)3-Amino-5-[6-(5-chloro-2-phenethyloxy-benzylamino)-pur-
in-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide.
C.sub.26H.sub.28ClN.sub.7O.sub.4. M.W.=538.01. M.S. 538
(M+H).sup.+. .sup.1H NMR (400 MHz, CD3OD) .delta. 8.35 (s, 1H);
8.27 (s, 1H); 7.3-7.05 (m, 7H); 6.92 (d, 1H, J=8.5 Hz); 6.05 (d,
1H, J=4.2 Hz); 5.45 (s, 2H); 4.65 (bs, 2H); 4.59 (m, 1H); 4.25 (d,
1H, J=5.4 Hz); 4.2 (dd, 2H, J=6.5, 6.0 Hz); 3.74 (m, 1H); 3.02 (dd,
2H, J=6.5, 6.0 Hz); 2.8 (s, 3H).
EXAMPLE 68
[0906]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(3,5-dimethyl-piperazin-1--
yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-ca-
rboxylic acid methylamide. C.sub.26H.sub.34ClN.sub.9O.sub.5 M.W.
588.1. MS 588 (M+H).sup.+.
[0907] .sup.1H NMR (400 MHz. CD.sub.3OD) .delta. 8.34 (s, 1H); 8.28
(s, 1H); 7.28 (s, 1H); 7.18 (dd, 1H, J=8.7, 2.6 Hz); 6.92 (d, 1H,
J=8.7 Hz); 6.06 (d, 1H, J=4.1 Hz); 4.81 (m, 2H); 4.60 (t, 1H, J=4.8
Hz); 4.4 (d, 1H, J=12 Hz); 4.31 (d, 1H, J=5.6 Hz); 3.83 (d, 1H,
12.9 Hz); 3.76 (m, 1H); 3.13 (m, 1H); 2.81 (s, 3H); 2.72 (m, 2H);
2.51 (s, 1H); 2.27 (t, 1H, J=12 Hz); 1.27 (s, 1H); 1.09 (d, 6H,
J=6.4 Hz).
EXAMPLE 69
[0908]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(4-dimethylamino-piperidin-
-1-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-
-carboxylic acid methylamide. C.sub.27H.sub.36ClN.sub.9O.sub.5 M.W.
602.09. MS 602 (M+H).sup.+. .sup.1H NMR (400 MHz. CD.sub.3OD)
.delta. 8.32 (s, 1H); 8.26 (s, 1H); 7.28 (s, 1H); 7.16 (dd, 1H,
J=8.7, 2.4 Hz); 6.90 (dd, 1H, J=8.7, 2.4 Hz); 6.06 (d,1H, J=4.0
Hz); 4.81 (s, 2H); 4.58 (t, 1H, J=4.8 Hz); 4.55 (d, 1H, J=12.0 Hz);
4.28 (d, 1H, J=5.4 Hz); 4.0 (d, 1H, J-13 Hz); 3.72 (s, 1H); 3.05
(t, 1H, J=5.7 Hz); 2.79 (s, 3H); 2.62 (t, 1H, J=13 Hz); 2.39 (m,
1H); 2.21 (s, 6H); 1.87 (s, 2H); 1.4-1.25 (m, 4H).
EXAMPLE 70
[0909]
(2S,3S,4R,5R)5-(6-{2-[2-(4-Adamantan-2-yl-piperazin-1-yl)-2-oxo-eth-
oxy]-5-chloro-benzylamino}-purin-9-yl)-3-amino-4-hydroxy-tetrahydro-furan--
2-carboxylic acid methylamide. C.sub.34H.sub.44ClN.sub.9O.sub.5
M.W. 694.24. MS 694 (M+H).sup.+.
[0910] .sup.1H NMR (400 MHz. CD.sub.3OD) .delta. 8.35 (s, 1H); 8.29
(s, 1H); 7.31 (d, 1H, J=2.1 Hz); 7.2 (dd, 1H, J=8.7, 2.6 Hz); 6.93
(d, 1H, J=8.9 Hz); 6.06 (d, 1H, J=4.3 Hz); 4.83 (m, 2H); 4.58 (t,
1H, J=4.8 Hz); 4.3 (d, 1H, J=5.6 Hz); 3.74 (t, 1H, 5.48 Hz); 3.59
(t, 4H, 4.87 Hz); 2.82 (s, 3H); 2.4 (s, 4H), 2.1-1.27 (m, 17H).
EXAMPLE 71
[0911]
(2R,3R,4S,5S)1-[(2-{[9-(4-Amino-3-hydroxy-5-methylcarbamoyl-tetrahy-
dro-furan-2-yl)-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetyl]-pipe-
ridine-4-carboxylic acid amide. C.sub.26H.sub.32ClN.sub.9O.sub.6
M.W. 602.05. MS 602 (M+H).sup.+. .sup.1H NMR (400 MHz. CD.sub.3OD)
.delta. 8.33 (s, 1H); 8.29 (s, 1H); 7.29 (s, 1H); 7.18 (dd, 1H,
J=8.7, 2.6 Hz); 6.93 (d, 1H, J=8.9 Hz); 6.06 (d, 1H, J=4.3 Hz);
4.90 (s, 3H); 4.61 (t, 1H, J=4.3 Hz); 4.4 (d, 1H); 4.3 (d, 1H,
J=5.6 Hz); 4.05 (d, 1H); 3.76 (t, 1H); 3.28 (m, 1H); 2.81 (s, 3H);
2.75 (m, 1H); 2.5 (m, 1H); 1.83 (d, 2H); 1.63 (m, 3H).
EXAMPLE 72
[0912]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(4-cycloheptyl-piperazin-1-
-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-c-
arboxylic acid methylamide. C.sub.31 H.sub.42 Cl N.sub.9 O.sub.5
M.W. 656.19. CIMS 656.1 (M+H).sup.+. .sup.1H NMR (400 MHz.
CD.sub.3OD) .delta.8.31 (s, 1H); 8.29 (s, 1H); 7.31 (s, 1H); 7.20
(dd, 1H, J=8.71, 2.69 Hz); 6.94 (d, 1H, J=8.71 Hz); 6.08 (d, 1H,
J=3.74 Hz); 4.90 (s, 2H); 4.82 (s, 2H); 4.72 (t, 1H J=4.56 Hz);
4.42 (d, 1H, J=6.02 Hz); 4.03 (t, 1 H, J=4.56 Hz); 3.62 (s, 4H);
2.78 (s, 3H); 2.66 (s, 5H); 1.81-1.38 (m, 12H)
EXAMPLE 73
[0913]
(2S,3S,4R,5R)5-{6-[2-(Adamantan-2-ylcarbamoylmethoxy)-5-chloro-benz-
ylamino]-purin-9-yl}-3-amino-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide.
[0914] C.sub.30H.sub.37Cl N.sub.8O.sub.5 M.W. 625.13. CIMS 625.0
(M+1). .sup.1H NMR (400 MHz. CD.sub.3OD) .delta. 8.36 (s, 1H); 8.28
(s, 1H); 7.34 (d, 1H, J=2.49 Hz); 7.24 (dd, 1H, J=8.72, 2.49 Hz);
6.94 (d, 1H, J=8.93 Hz); 6.06 (d, 1H, J=4.15); 4.83 (s, 2H); 4.64
(s 2H); 4.58 (t, 1H, J=4.77 Hz), 4.30 (d, 1H, J=5.61 Hz); 4.01 (s,
1H); 3.74 (t, 1H, 5.61 Hz); 2.81 (s, 3H); 1.81-1.43 (m, 14H).
EXAMPLE 74
[0915]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(1-phenyl-ethoxy)-benzylamino-
]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide. C.sub.26H.sub.28ClN.sub.7O.sub.4. M.W.=538.01. M.S.
538 (M+H).sup.+. .sup.1H NMR (400 MHz, CD30D) .delta.8.37 (s, 1H);
8.25 (s, 1H); 7.42-7.08 (m, 6H); 7.0 (d, 1H, J=8.5 Hz); 6.74 (d,
1H, J=8.5 Hz); 6.05 (d, 1H, J=4.2 Hz); 5.4 (q, 1H, J=6.8 Hz); 4.82
(bs, 2H); 4.59 (m, 1H); 4.3 (d, 1H, J=5.5 Hz); 3.73 (m, 1H); 2.8
(s, 3H); 1.58 (bs, 3H).
EXAMPLE 75
[0916]
(2R,3R,4S,5S)4-[(2-{[9-(4-Amino-3-hydroxy-5-methylcarbamoyl-tetrahy-
dro-furan-2-yl)-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetyl]-pipe-
razine-1-carboxylic acid tert-butyl ester.
C.sub.29H.sub.38ClN.sub.9O.sub.- 7 M.W. 660.13. MS 660 (M+H).sup.+.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33 (s, 1H); 8.28 (bs,
1H); 7.3 (bs, 1H); 7.2 (d, 1H, J=8.9 Hz); 6.93 (d, 1H, J=8.9 Hz);
6.07 (d, 1H, J=3.9 Hz); 4.91 (s, 2H); 4.82 (bs, 2H); 4.6 (t, 1H,
J=4.5 Hz); 4.3 (d, 1H, J=5.6 Hz); 3.81 (t, 1H, J=5.0 Hz); 3.6 (m,
4H); 3.42 (m, 4H); 2.8 (s, 3H); 1.42 (s, 9H).
EXAMPLE 76
[0917]
(2S,3S,4R,5R)3-Amino-5-[6-(5-chloro-2-cyanomethoxy-benzylamino)-pur-
in-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide.
C.sub.20H.sub.21 ClN.sub.8O.sub.4 M.W.472.89. MS 473
(M+H).sup.+.
[0918] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.36 (s, 1H); 8.29
(s, 1H); 7.32 (bs, 1H); 7.28 (dd, 1H, J=8.9, 2.5 Hz); 7.08 (d, 1H,
J=8.9 Hz); 6.07 (d, 1H, J=4.1 Hz); 5.05 (s, 2H); 4.8 (bs, 2H); 4.6
(t, 1H, J=4.9 Hz); 4.3 (d, 1H, J=5.6 Hz); 3.78 (t, 1H, J=5.5 Hz);
2.8 (s, 3H).
EXAMPLE 77
[0919]
(2R,3R,4S,5S)(2-{[9-(4-Amino-3-hydroxy-5-methylcarbamoyl-tetrahydro-
-furan-2-yl-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetic
acid methyl ester. C.sub.21H.sub.38ClN.sub.7O.sub.6 M.W.505.92. MS
506 (M+H).sup.+.
[0920] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.35 (s, 1H); 8.29
(s, 1H); 7.3 (bs, 1H); 7.2 (dd, 1H, J=8.9, 2.5 Hz); 6.9 (d, 1H,
J=8.9 Hz); 6.04 (d,1H, J=4.0 Hz); 4.8 (m, 4H); 4.6 (t, 1H, J=4.5
Hz); 4.3 (d, 1H, J=5.6 Hz); 3.78 (m, 4H); 2.8 (s, 3H).
EXAMPLE 78
[0921]
(2R,3R,4S,5S)(2-{[9-(4-Amino-3-hydroxy-5-methylcarbamoyl-tetrahydro-
-furan-2-yl-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetic
acid ethyl ester.
[0922] C.sub.22H.sub.26ClN.sub.7O.sub.6 M.W.519.95. MS 520
(M+H).sup.+.
[0923] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.35 (s, 1H); 8.29
(s, 1H); 7.3 (bs, 1H); 7.2 (dd, 1H, J=8.9, 2.5 Hz); 6.9 (d, 1H,
J=8.9 Hz); 6.04 (d, 1H, J=4.0 Hz); 4.82 (s, 2H); 4.8 (s, 2H); 4.6
(t, 1H, J=4.5 Hz); 4.3 (d, 1H, J=5.6 Hz); 4.21 (q, 2H, J=7.1 Hz);
3.78 (t, 1H, J=5.4 Hz); 2.8 (s, 3H); 1.22 (t, 3H, J=7.1 Hz).
EXAMPLE 79
[0924]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(4,5-dihydro-1H-imidazol-2-yl-
methoxy)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide. C.sub.22H.sub.26ClN.sub.9O.sub.4 M.W.515.96. MS
516 (M+H).sup.+.
[0925] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.4 (s, 1H); 8.3
(s, 1H); 7.32 (bs, 1H); 7.2 (d, 1H, J=8.9 Hz); 6.97 (d, 1H, J=8.9
Hz); 6.05 (d, 1H, J=3.8 Hz); 4.84 (m, 4H); 4.6 (t, 1H, J=4.5 Hz);
4.3 (d, 1H, J=5.6 Hz); 3.75 (t, 1H, J=5.4 Hz); 3.62 (m, 4H); 2.8
(s, 3H).
EXAMPLE 80
[0926]
(2S,3S,4R,5R)3-Amino-5-{6-[1-(2-benzyloxy-5-chloro-phenyl)-ethylami-
no]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide. C.sub.26H.sub.28ClN.sub.7O.sub.4.
[0927] M.W.=538.01. M.S. 538 (M+H).sup.+. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 8.39 (bs, 1H); 8.2 (d, 1H, J=4.5 Hz); 7.5-7.22
(m, 5H); 7.18 (d, 1H, J=8.9 Hz); 7.0 (d, 2H, J=8.6 Hz); 6.05 (d,
1H, J=3.9 Hz); 5.79 (bs, 1H); 5.18 (s, 2H); 4.6 (t, 1H, J=4.5 Hz);
4.3 (d, 1H, J=5.6 Hz); 3.7 (m, 1H); 2.8 (s, 3H); 1.6 (d, 3H, J=6.8
Hz).
EXAMPLE 81
[0928]
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzyloxy-5-chloro-benzylamino)-2-chlo-
ro-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide. C.sub.25H.sub.25Cl.sub.2N.sub.7O.sub.4.
[0929] M.W.=558.43. M.S. 558 (M+H).sup.+. .sup.1 H NMR (400 MHz,
D6DMSO) .delta. 8.82 (m, 1H); 8.6 (bs, 1H); 8.2 (bs, 1H); 7.43-7.02
(m, 7H); 5.95 (bs, 2H); 4.62 (bs, 2H); 4.3 (bs, 1H); 4.1 (m, 2H);
3.52 (m, 1H); 3.1 (d, 3H, J=5.0 Hz); 2.42 (bs, 2H); 1.75 (bs,
2H).
EXAMPLE 82
[0930]
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzylsulfanyl-5-chloro-benzylamino)-p-
urin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide. C.sub.25H.sub.26ClN.sub.7O.sub.4S. M.W.=540.05. M.S.
540 (M+H).sup.+. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.35 (s,
1H); 8.27 (s, 1H); 7.4-7.1 (m, 8H); 6.07 (d, 1H, J=3.9 Hz); 4.8
(bs, 2H); 4.61 (m, 1H); 4.35 (d, 1H, J=5.8 Hz); 4.08 (s, 2H); 3.82
(m, 1H); 2.8 (s, 3H).
EXAMPLE 83
[0931]
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzyloxy-5-bromo-benzylamino)-purin-9-
-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide.
C.sub.25H.sub.26BrN.sub.7O.sub.4. M.W.=568.43. M.S. 568
(M+H).sup.+. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.35 (s,
1H); 8.25 (s, 1H); 7.42-7.23 (m, 7H); 6.97 (d, 1H, J=9.3 Hz); 6.06
(d, 1H, J=4.1 Hz); 5.13 (s, 2H); 4.8 (bs, 2H); 4.6 (m, 1H); 4.3 (d,
1H, J=5.5 Hz); 3.79 (m, 1H); 2.8 (s, 3H).
EXAMPLE 84
[0932]
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzyloxy-5-fluoro-benzylamino)-purin--
9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide.
C.sub.25H.sub.26FN.sub.7O.sub.4. M.W.=507.53. M.S. 508 (M+H).sup.+.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.32 (s, 1H); 8.26 (s,
1H); 7.42-7.22 (m, 4H); 7.1-6.9 (m, 4H); 6.06 (d, 1H, J=4.2 Hz);
5.13 (s, 2H); 4.8 (bs, 2H); 4.5 (m, 1H); 4.3 (d, 1H, J=5.5 Hz);
3.79 (m, 1H); 2.8 (s, 3H).
EXAMPLE 85
[0933]
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzyloxy-5-iodo-benzylamino)-purin-9--
yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide.
C.sub.25H.sub.26IN.sub.7O.sub.4. M.W.=615.43. M.S. 490 (M-I).sup.+.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.32 (s, 1H); 8.21 (s,
1H); 7.42-7.18 (m, 5H); 7.02-6.82 (m, 3H); 6.05 (d, 1H, J=4.2 Hz);
5.1 (d, 2H, J=4.9 Hz); 4.83 (bs, 2H); 4.55 (m, 1H); 4.29 (d,1H,
J=5.5 Hz); 3.7 (m, 1H 2.8 (s, 3H).
EXAMPLE 86
[0934]
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzyloxy-5-trifluoromethyl-benzylamin-
o)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide. C.sub.26H.sub.26F.sub.3N.sub.7O.sub.4.
[0935] M.W.=557.54. M.S. 558 (M+H).sup.+. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 8.36 (s, 1H); 8.24 (s, 1H); 7.6-7.18 (m, 8H);
6.06 (d, 1H, J=4.1 Hz); 5.21 (s, 2H); 4.83 (bs, 2H); 4.6 (m, 1H);
4.3 (d, 1H, J=5.7 Hz); 3.75 (m, 1H); 2.8 (s, 3H).
EXAMPLE 87
[0936]
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzyloxy-5-cyano-benzylamino)-purin-9-
-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide.
C.sub.26H.sub.26N.sub.8O.sub.4. M.W.=514.55. M.S. 515 (M+H).sup.+.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.36 (s, 1H); 8.24 (s,
1H); 7.63-7.2 (m, 8H); 6.06 (d, 1H, J=4.1 Hz); 5.24 (s, 2H); 4.83
(bs, 2H); 4.6 (m, 1H); 4.32 (d, 1H, J=5.4 Hz); 3.76 (m, 1H); 2.8
(s, 3H).
EXAMPLE 88
[0937]
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzyloxy-5-methyl-benzylamino)-purin--
9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide.
C.sub.26H.sub.29N.sub.7O.sub.4. M.W.=503.56. M.S. 504 (M+H).sup.+.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.31 (s, 1H); 8.22 (s,
1H); 7.4-6.9 (m, 8H); 6.03 (d, 1H, J=4.3 Hz); 5.04 (s, 2H); 4.8
(bs, 2H); 4.58 (m, 1H); 4.3 (d, 1H, J=5.6 Hz); 3.7 (m, 1H); 2.8 (s,
3H); 2.2 (s, 3H).
EXAMPLE 89
[0938]
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzyloxy-5-vinyl-benzylamino)-purin-9-
-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide.
C.sub.27H.sub.29N.sub.7O.sub.4. M.W.=515.57. M.S. 516 (M+H).sup.+.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.33 (s, 1H); 8.22 (s,
1H); 7.42-7.2 (m, 7H); 6.98 (d, 1H, J=8.5 Hz); 6.6 (dd, 1H, J=8.2,
6.0 Hz); 6.03 (d, 1H, J=4.2 Hz); 5.58 (d, 1H, J=8.2 Hz); 5.1 (s,
2H); 4.8 (bs, 2H); 4.58 (m, 1H); 4.26 (d, 1H, J=5.5 Hz); 3.72 (m,
1H); 2.8 (s, 3H).
EXAMPLE 90
[0939]
(2S,3S,4R,5R)3-Amino-5-[6-(2-benzyloxy-5-ethynyl-benzylamino)-purin-
-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide.
C.sub.27H.sub.27N.sub.7O.sub.4. M.W.=513.56. M.S. 514 (M+H).sup.+.
.sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.36 (s, 1H); 8.26 (s,
1H); 7.7-7.23 (m, 7H); 7.0 (d, 1H, J=8.5 Hz); 6.06 (d, 1H, J=4.2
Hz); 5.18 (s, 2H); 4.8 (bs, 2H); 4.6 (m, 1H); 4.3 (d, 1H, J=5.5
Hz); 3.75 (m, 1H); 3.3 (s, 1H); 2.8 (s, 3H).
EXAMPLE 91
[0940]
(2S,3S,4R,5R)3-Amino-5-{6-[5-chloro-2-(2-oxo-2-(4-piperidin-1-yl-et-
hoxy)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide. C.sub.30H.sub.40ClN.sub.9O.sub.5 M.W. 642.18. MS
642 (M+H).sup.+.
[0941] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.4 (s, 1H); 8.32
(bs, 1H); 7.35 (bs, 1H); 7.22 (d, 1H, J=8.9 Hz); 6.96 (d, 1H, J=8.9
Hz); 6.07 (d, 1H, J=4.1 Hz); 4.91 (s, 2H); 4.82 (bs, 2H); 4.6 (m,
2H); 4.33 (d, 1H, J=5.6 Hz); 4.2 (bd, 1H, J=10.5 Hz); 3.79 (t, 1H,
J=5.4 Hz); 3.18 (m, 1H); 2.82 (s, 3H);. 2.6 (m, 6H); 1.9 (m, 2H);
1.65-1.3 (m, 8H).
EXAMPLE 92
[0942]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(4-methylamino-piperidin-1-
-yl-)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2--
carboxylic acid methylamide. C.sub.26H.sub.34ClN.sub.9O.sub.5 M.W.
588.07. MS 588 (M+H).sup.+. .sup.1H NMR (400 MHz. CD.sub.3OD)
.delta. 8.39 (s, 1H); 8.33 (s, 1H); 7.35 (s, 1H); 7.22 (dd, 1H,
J=8.7, 2.4 Hz); 6.94 (dd, 1H, J=8.7, 2.4 Hz); 6.06 (d, 1H, J=4.0
Hz); 4.95 (s, 2H); 4.81 (s, 2H); 4.62 (t, 1H, J=4.8 Hz); 4.5 (bd,
1H, J=12.0 Hz); 4.36 (d, 1H, J=5.4 Hz); 4.02 (bd, 1H, J=13 Hz);
3.79 (t, 1H, J=5.7 Hz); 3.2 (m, 1H); 2.82 (s, 3H); 2.79 (t, 1H,
J=13 Hz); 2.62 (m, 1H); 2.4 (s, 3H); 1.97 (m, 2H); 1.4-1.15 (m,
2H).
EXAMPLE 93
[0943]
(2S,3S,4R,5R)3-Amino-5-(6-{5-chloro-2-[2-(4-amino-piperidin-1-yl)-2-
-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-carboxy-
lic acid methylamide. C.sub.25H.sub.32ClN.sub.9O.sub.5 M.W. 574.04.
MS 574 (M+H).sup.+.
[0944] .sup.1H NMR (400 MHz. CD.sub.3OD) .delta. 8.37 (s, 1H); 8.28
(s, 1H); 7.3 (s, 1H); 7.2 (dd, 1H, J=8.7, 2.4 Hz); 6.92 (dd, 1H,
J=8.7, 2.4 Hz); 6.06 (d, 1H, J=4.0 Hz); 4.95 (s, 2H); 4.81 (s, 2H);
4.6 (t, 1H, J=4.8 Hz); 4.4 (bd, 1H, J=12.0 Hz); 4.3 (d, 1H, J=5.4
Hz); 3.95 (bd, 1H, J=13 Hz); 3.75 (t, 1H, J=5.7 Hz); 3.1 (m, 1H);
2.8 (s, 3H); 2.79 (m, 2H); 2.62 (m, 1H); 1.82 (m, 2H); 1.4-1.15 (m,
2H).
Preparation A1
BOC Cleavage
[0945]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(2-oxo-2-piperazin-1-yl-ethox-
y)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide To trifluoroacetic acid (30 mL) was added to
(2S,3S,4R,5R)3-azido-5-{6-[5-chloro-2-(2-oxo-2-(4-tert-butyloxycarbonyl)--
piperazin-1-yl-ethoxy)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-
-2-carboxylic acid methylamide (3 g, 4.5 mmol) at ambient
temperature. After 1 hour, the mixture was concentrated, and the
residue reconcentrated from chloroform three times. The crude
product was dissolved in methanol (80 mL) and dichloromethane (100
mL) and neutralized with Amberlite IR 400 (OH) resin. The mixture
was filtered and concentrated and the residue was purified by
passing through a plug of silica gel (7.5-10%
[0946] methanol/dichloromethane/0.1% NH.sub.4OH) to afford 2.24 g
of the title compound as a colorless solid.
[0947] In an analogous manner the following compounds, preparations
A2-A3, were prepared from the appropriate protected amine using
analogous procedures to Preparation A1.
Preparation A2
[0948]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(4-amino-piperidin-1-yl)-2-
-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-carboxy-
lic acid methylamide.
Preparation A3
[0949]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(4-methylamino-piperidin-1-
-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-c-
arboxylic acid methylamide.
Preparation B1
Amide Coupling
[0950]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-oxo-2-(3-oxo-piperazin-1-y-
l)-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-carboxyli-
c acid methylamide
[0951] EDCl (44 mg, 0.23 mmol), HOBT (30 mg, 0.22 mmol) and DMAP
(40 mg) were added to a mixture of
(2R,3R,4S,5S)(2-{[9-(4-Azido-3-hydroxy-5-methy-
lcarbamoyl-tetrahydro-furan-2-yl)-9H-purin-6-ylamino]-methyl}-4-chloro-phe-
noxy)-acetic acid (60 mg, 0.116 mmol) and piperizin-2-one (35 mg,
0.35 mmol) in anhydrous DMF (3 mL). After 20 hours at room
temperature, the mixture was concentrated, pre-adsorbed onto silica
gel and purified by flash chromatography (6-8%
methanol/dichloromethane) to afford the title compound as a
colorless solid. MS: 600 (M+H).sup.+.
[0952] In an analogous manner the following compounds, preparations
B2-B38, were prepared from the appropriate amine using analogous
procedures to the Preparation B1.
Preparation B2
[0953]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(2-oxo-2-piperidin-1-yl-ethox-
y)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide.
Preparation B3
[0954]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-phenylcarbamoylmethoxy-benzyl-
amino)-purin-9-yl]4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide.
Preparation B4
[0955]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-dimethylcarbamoylmethoxy-benz-
ylamino)-purin-9-yl]4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide
Preparation B5
[0956]
(2S,3S,4R,5R)3-Azido-5-{6-[2-(benzylcarbamoyl-methoxy)-5-chloro-ben-
zylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide.
Preparation B6
[0957]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(4-methyl-piperazin-1-yl)--
2-oxo-ethoxy]-benzylamino}-purin-9-yl)4-hydroxy-tetrahydro-furan-2-carboxy-
lic acid methylamide.
Preparation B7
[0958]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-propylcarbamoylmethoxy-benzyl-
amino)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide.
Preparation B8
[0959]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(2-morpholin-4-yl-2-oxo-ethox-
y)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide.
Preparation B9
[0960]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(2-oxo-2-pyrrolidin-1-yl-etho-
xy)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide
Preparation B10
[0961]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-dipropylcarbamoylmethoxy-benz-
ylamino)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide
Preparation B11
[0962]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[(2-methoxy-ethylcarbamoyl)-m-
ethoxy]-benzylamino}-purin-9-yl)4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide.
Preparation B12
[0963]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-methylcarbamoylmethoxy-benzyl-
amino)-purin-9-yl]4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide.
Preparation B13
[0964]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-cyclohexylcarbamoylmethoxy-be-
nzylamino)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide
Preparation B14
[0965]
(2R,3R,4S,5S)4-[(2-{[9-(4-Azido-3-hydroxy-5-methylcarbamoyl-tetrahy-
dro-furan-2-yl)-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetyl]-pipe-
razine-1-carboxylic acid ethyl ester
Preparation B15
[0966]
(2S,3S,4R,5R)3-Azido-5-{6-[2-(2-azetidin-1-yl-2-oxo-ethoxy)-5-chlor-
o-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide
Preparation B16
[0967]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[(2-morpholin4-yl-ethylcarbam-
oyl)-methoxy]-benzylamino}-purin-9-yl)4-hydroxy-tetrahydro-furan-2-carboxy-
lic acid methylamide
Preparation B17
[0968]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-oxo-2-(4-phenyl-piperazin--
1-yl)-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-carbox-
ylic acid methylamide
Preparation B18
[0969]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(4-cyclohexyl-piperazin-1--
yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-ca-
rboxylic acid methylamide
Preparation B19
[0970]
(2S,3S,4R,5R)3-Azido-5-(6-(5-chloro-2-[2-(4-ethyl-piperazin-1-yl)-2-
-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-carboxy-
lic acid methylamide.
Preparation B20
[0971]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-cyclopropylcarbamoylmethoxy-b-
enzylamino)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide
Preparation B21
[0972]
(2S,3S,4R,5R)3-Azido-5-[6-(2-carbamoylmethoxy-5-chloro-benzylamino)-
-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide
Preparation B22
[0973]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(4-cyclopropyl-piperazin-1-
-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-c-
arboxylic acid methylamide
Preparation B23
[0974]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(4-isopropyl-piperazin-1-y-
l)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)4-hydroxy-tetrahydro-furan-2-carb-
oxylic acid methylamide
Preparation B24
[0975]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-oxo-2-(4-propyl-piperazin--
1-yl)-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-carbox-
ylic acid methylamide
Preparation B25
[0976]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(4-cyclopentyl-piperazin-1-
-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)4-hydroxy-tetrahydro-furan-2-ca-
rboxylic acid methylamide
Preparation B26
[0977]
(2S,3S,4R,5R)3-Azido-5-(6-{2-[2-(4-benzyl-piperazin-1-yl)-2-oxo-eth-
oxy]-5-chloro-benzylamino}-purin-9-yl)4-hydroxy-tetrahydro-furan-2-carboxy-
lic acid methylamide
Preparation B27
[0978]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(4-ethyl-3-oxo-piperazin-1-
-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-c-
arboxylic acid methylamide
Preparation B28
[0979]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-{2-[4-(2-chloro-phenyl)-piper-
azin-1-yl]-2-oxo-ethoxy}-benzylamino)-purin-9-yl]-4-hydroxy-tetrahydro-fur-
an-2-carboxylic acid methylamide
Preparation B29
[0980]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(phenethylcarbamoyl-methoxy)--
benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide
Preparation B30
[0981]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(3,5-dimethyl-piperazin-1--
yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-ca-
rboxylic acid methylamide
Preparation B31
[0982]
(2S,3S,4R,5R)3-Azido-5-(6){5-chloro-2-[2-(4-dimethylamino-piperidin-
-1-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-
-carboxylic acid methylamide
Preparation B32
[0983]
(2S,3S,4R,5R)5-(6-{2-[2-(4-Adamantan-2-yl-piperazin-1-yl)-2-oxo-eth-
oxy]-5-chloro-benzylamino}-purin-9-yl)-3-azido4-hydroxy-tetrahydro-furan-2-
-carboxylic acid methylamide
Preparation B33
[0984]
(2R,3R,4S,5S)1-[(2-{[9-(4-Azido-3-hydroxy-5-methylcarbamoyl-tetrahy-
dro-furan-2-yl)-9H-purin-6-ylamino]-methyl}4-chloro-phenoxy)-acetyl]-piper-
idine4-carboxylic acid amide
Preparation B34
[0985]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(4-cycloheptyl-piperazin-1-
-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-c-
arboxylic acid methylamide
Preparation B35
[0986]
(2S,3S,4R,5R)5-{6-[2-(Adamantan-2-ylcarbamoylmethoxy)-5-chloro-benz-
ylamino]-purin-9-yl}-3-azido-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide
Preparation B36
[0987]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(2-oxo-2-(4-tert-butyloxycarb-
onyl)-piperazin-1-yl-ethoxy)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-
-furan-2-carboxylic acid methylamide
Preparation B37
[0988]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(4-tert-butyloxycarbonylam-
ino-piperiden-1-yl)-2-oxo-ethoxy]-benzylamino)-purin-9-yl)-4-hydroxy-tetra-
hydro-furan-2-carboxylic acid methylamide.
Preparation B38
[0989]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[2-(4-tert-butyloxycarbonyl-m-
ethylamino-piperidin-1-yl)-2-oxo-ethoxy]-benzylamino}-purin-9-yl)-4-hydrox-
y-tetrahydro-furan-2-carboxylic acid methylamide.
Preparation C1
Deprotection of t-Butyl Ester
[0990]
(2R,3R,4S,5S)(2-{[9-(4-Azido-3-hydroxy-5-methylcarbamoyl-tetrahydro-
-furan-2-yl)-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetic
acid
[0991]
(2R,3R,4S,5S)(2-{[9-(4-Azido-3-hydroxy-5-methylcarbamoyl-tetrahydro-
-furan-2-yl)-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetic
acid tert-butyl ester (1 g, 1.9 mmol) was added to trifluoroacetic
acid (15 mL) and stirred at room temperature for 3 hours. The
mixture was concentrated and the residue was reconcentrated from
chloroform three times to afford the title compound as a foam. M.S.
518
[0992] (M+H).sup.+. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.5
(m, 2H); 7.38 (bs, 1H); 7.25 (d, 1H, J=8.0 Hz); 6.95 (d, 1H, J=8.0
Hz); 6.0 (d, 1H J=4.0 Hz); 5.0 (m, 1H); 4.85 (m, 2H); 4.75 (s, 2H);
4.4 (m, 2H); 2.8 (s, 3H).
Preparation D1
Alkylation of Phenol
[0993]
(2R,3R,4S,5S)(2-{[9-(4-Azido-3-hydroxy-5-methylcarbamoyl-tetrahydro-
-furan-2-yl)-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetic
acid ethyl ester
[0994] Sodium hydride (115 mg, 4.8 mmol) was added to a solution of
(2S,3S,4R,5R)3-azido-5-{6-[5-chloro-2-hydroxy-benzylamino]-purin-9-yl}-4--
hydroxy-tetrahydro-furan-2-carboxylic acid methylamide (2.06 g,
4.37 mmol) in anhydrous DMF (30 mL) while cooled in an ice bath.
After 30 minutes, ethyl bromoacetate (0.58 mL, 5.25 mmol) was added
and the reaction was warmed to ambient temperature and stirred
overnight. The reaction was quenched with methanol and
concentrated. The residue was preadsorbed on silica gel and
purified by flash chromatography (2-6% methanol/dichloromethane) to
afford 1.7 g product as a colorless solid. MS 546
[0995] (M+H).sup.+. .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 8.34
(bs, 1H); 8.3 (s, 1H); 7.32 (bs, 1H); 7.22 (dd, 1H, J=8.9, 2.6 Hz);
6.9 (d, 1H, J=8.9 Hz); 6.02 (d, 1H, J=7.1 Hz); 5.07 (dd, 1H, J=7.1,
4.9 Hz); 4.85 (bs, 2H); 4.8 (s, 2H); 4.42 (m, 2H); 4.25 (q, 2H,
J=6.9 Hz); 3.0 (s, 1H); 2.85 (s, 3H); 1.28 (t, 3H, J=6.9 Hz).
[0996] In an analogous manner the following compounds, preparations
D2-D3, were prepared from the appropriate starting material using
analogous procedures to Preparation D1.
Preparation D2
[0997]
(2R,3R,4S,5S)(2-{[9-(4-Azido-3-hydroxy-5-methylcarbamoyl-tetrahydro-
-furan-2-yl)-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetic
acid methyl ester
Preparation D3
[0998]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-cyanomethoxy-benzylamino)-pur-
in-9-yl]-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide
Preparation E1
[0999]
(2S,3S,4R,5R)3-Azido-5-6-[5-chloro-2-(2-morpholin-4-yl-ethoxy)-benz-
ylamino]-purin-9-y})-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1000] To a mixture of
4-acetoxy-3-azido-5-(6-chloropurin-9-yl)tetrahydrof-
uran-2-carboxylic acid methyl ester (485 mg, 1.27 mmol) and in
anhydrous methanol (20 mL) was added triethylamine (0.51 mL, 3.8
mmol) and the reaction was heated to 50.degree. C. under anhydrous
conditions. The reaction was stirred at reflux for 15 hours.
Methylamine (3.85 mL, 1.0 M in MeOH) was added to the reaction and
the reaction was stirred at room temperature for another 15 hours.
The solvent was then removed by rotary evaporation and the product
was preadsorbed onto silica gel and purified by flash
chromatography (SiO.sub.2, 2.5% MeOH, EtoAc) to afford the title
compound.
[1001] C.sub.24H.sub.29ClN.sub.10O.sub.5. MW 573.02. MS 573.1
(M+H).sup.+.
[1002] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.64 (quart, 1H,
J=4.6 Hz); 8.46 (s, 1H); 8.35 (bs, 1H); 8.23 (s, 1H); 7.21 (dd, 1H,
J=8.7 Hz, J=2.7 Hz); 7.05 (bs, 1H); 7.01 (d, 1H, J=8.7 Hz); 6.30
(d, 1H, J=5.6 Hz); 5.98 (d, 1H, J=6.2 Hz) 4.98 (quart, 1H, J=5.5
Hz); 4.64 (bs, 2H); 4.50-4.45 (mult, 1H); 4.31 (d, 1H, J=3.1 Hz);
4.12 (t, 2H, J=5.7 Hz); 3.57-3.51 (mult, 4H); 2.72-2.62 (mult, 5H);
2.51-2.45 (mult, 4H).
[1003] The following compounds, Preparation E2-E18, were prepared
by analogous procedures to Preparation E1 using three
propatedbenzyl amine.
Preparation E2
[1004]
(2S,3S,4R,5R)3-Azido-5-[6-(2-benzyloxy-5-chloro-benzylamino)-purin--
9-yl]-4-hydroxytetrahydrofuran-2-carboxylic acid methylamide
[1005] C.sub.25H.sub.24ClN.sub.9O.sub.4. MW 549.98. MS 550.1
(M+H).sup.+.
[1006] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.65 (quart, 1H,
J=4.6 Hz); 8.47 (s, 1H); 8.42 (bs, 1H); 8.23 (s, 1H); 7.49-7.45
(mult, 2H);7.40 to 7.35 (mult, 2H); 7.30 (t, 1H, J=7.1 Hz); 7.21
(dd, 1H, J=8.7 Hz, J=2.5 Hz); 7.20 (d, 2H, J=8.9 Hz); 6.31 (d, 1H,
J=5.4 Hz); 5.98 (d, 1H, J=6.4 Hz); 5.17 (s, 2H); 4.96 (quart, 1H,
J=5.8 Hz); 4.71 (bs, 2H); 4.52-4.45 (mult, 1H); 4.35-4.30 (mult,
1H); 2.66 (d, 3H, J=4.6 Hz).
Preparation E3
[1007]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-cyclobutylmethoxybenzylamino)-
-purin-9-yl]-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1008] C.sub.23H.sub.26ClN.sub.9O.sub.4. MW 527.97. MS 527.7
(M+H).sup.+.
[1009] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.65 (quart, 1H,
J=4.6 Hz); 8.47 (s, 1H); 8.35 (bs, 1H); 8.23 (s, 1H); 7.20 (dd, 1H,
J=8.7 Hz, J=2.7 Hz); 7.03 (bs, 1H); 6.98 (d, 1H, J=8.7 Hz); 6.31
(d, 1H, J=5.0 Hz); 5.98 (d, 1H, J=6.4 Hz); 4.96 (quart, 1H, J=5.2
Hz); 4.64 (bs, 2H); 4.48 (bs, 1H); 4.30-4.32 (mult, 1H); 3.97 (d,
2H, J=6.2 Hz); 2.71 (bs, 1H); 2.66 (d, 3H, J=4.6 Hz); 2.03 (bs,
2H); 1.86 (bs, 4H).
Preparation E4
[1010]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(3-methoxy-benzyloxy)-benzyla-
mino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1011] C.sub.26H.sub.26ClN.sub.9O.sub.5. MW 580.01. MS 579.8
(M+H).sup.+.
[1012] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.65 (quart, 1H,
J=4.6 Hz); 8.47 (s, 1H); 8.43 (bs, 1H); 8.24 (s, 1H); 7.28 (t, 1H,
J=8.2 Hz); 7.21 (dd, 1H, J=8.6 Hz, J=2.5 Hz); 7.10-7.00 (mult, 4H);
6.86 (d, 1H, J=6.8 Hz); 6.31 (d, 1H, J=5.4 Hz); 5.98 (d, 1H, J=6.4
Hz); 5.15 (s, 2H); 4.96 (quart, 1H, J=5.5 Hz); 4.71 (bs, 2H);
4.50-4.45 (mult, 1H); 4.31 (d, 1H, J=2.9 Hz); 3.72 (s, 3H); 2.66
(d, 3H, J=4.6 Hz).
Preparation E5
[1013]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(2,5-dimethoxy-benzyloxy)-ben-
zylamino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1014] C.sub.27H.sub.28ClN.sub.9O.sub.6. MW 610.03. MS 609.9
(M+H).sup.+.
[1015] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.66 (quart, 1H,
J=4.8 Hz); 8.47 (s, 1H); 8.41 (bs, 1H); 8.23 (s, 1H); 7.21 (dd, 1H,
J=8.7 Hz, J=2.7 Hz); 7.07 (bs, 1H); 7.05-7.00 (mult, 2H); 6.96 (d,
1H, J=8.9 Hz); 6.86 (dd, 1H, J=8.9 Hz, J=3.1 Hz); 6.31 (d, 1H,
J=5.4 Hz); 5.98 (d, 1H, J=6.4 Hz); 5.10 (s, 2H); 4.95 (quart, 1H,
J=5.8 Hz); 4.69 (bs, 2H); 4.49-4.48 (mult, 1H); 4.31 (d, 1H, J=2.9
Hz); 3.77 (s, 3H); 3.75 (s, 3H); 2.66 (d, 3H, J=4.6 Hz).
Preparation E6
[1016]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(3-chloro-benzyloxy)-benzylam-
ino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1017] Mp 89.0-95.0.degree. C.
[1018] C.sub.25H.sub.23Cl.sub.2N.sub.9O.sub.4. MW 584.43. MS 583.8
(M+H).sup.+.
[1019] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.63 (quart, 1H,
J=4.4 Hz); 8.45 (s, 1H); 8.45 (bs, 1H); 8.23 (s, 1H); 7.52 (s, 1H);
7.45-7.32 (mult, 3H); 7.21 (dd, 1H, J=8.9 Hz, J=2.7 Hz); 7.10 (bs,
1H); 7.02 (d, 1H, J=8.7 Hz); 6.29 (d, 1H, J=5.2 Hz); 5.96 (d, 1H,
J=6.4 Hz); 5.17 (s, 2H); 4.98-4.90 (mult, 1H); 4.69 (bs, 2H);
4.48-4.44 (mult, 1H); 4.30 (d, 1H, J=2.9 Hz); 2.64 (d, 3H, J=4.4
Hz).
Preparation E7
[1020]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(4-chloro-benzyloxy)-benzylam-
ino]-purin-9-yl)-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1021] Mp 79.0-84.0.degree. C.
[1022] C.sub.25H.sub.23Cl.sub.2N.sub.9O.sub.4. MW 584.43. MS 584.1
(M+H).sup.+.
[1023] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.66 (quart, 1H,
J=4.4 Hz); 8.47 (s, 1H); 8.44 (bs, 1H); 8.24 (s, 1H); 7.50 (d, 2H,
J=8.3 Hz); 7.47-7.41 (mult, 2H); 7.25-7.20 (mult, 1H); 7.15-7.05
(mult, 2H); 6.32 (d, 1H, J=5.4 Hz); 5.99 (d, 1H, J=7.7 Hz); 5.17
(s, 2H); 4.96 (quart, 1H, J=5.2 Hz); 4.70 (bs, 2H); 4.49 (bs, 1H);
4.32 (d, 1H, J=2.9 Hz); 2.67 (d, 3H, J=4.4 Hz).
Preparation E8
[1024]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(2-chloro-benzyloxy)-benzylam-
ino]-purin-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1025] Mp 66.0-70.0.degree. C.
[1026] C.sub.25H.sub.23Cl.sub.2N.sub.9O.sub.4. MW 584.43. MS 584.1
(M+H).sup.+.
[1027] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.66 (quart, 1H,
J=4.8 Hz); 8.47 (s, 1H); 8.42 (bs, 1H); 8.22 (s, 1H); 7.67-7.61
(mult, 1H); 7.52-7.48 (mult, 1H); 7.41-7.34 (mult, 2H); 7.27-7.18
(mult, 1H); 7.11 (d, 2H, J=9.1 Hz); 6.32 (d, 1H, J=5.4 Hz); 5.98
(d, 1H, J=6.4 Hz); 5.22 (s, 2H); 4.96 (quart, 1H, J=5.8 Hz); 4.70
(bs, 2H); 4.50-4.45 (mult, 1H); 4.32 (d, 1H, J=3.1 Hz); 2.66 (d,
3H, J=4.8 Hz).
Preparation E9
[1028]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(tetrahydrofuran-3-ylmethoxy)-
-benzylamino]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic
acid methylamide
[1029] C.sub.23H.sub.26ClN.sub.9O.sub.5. MW 543.97. MS 544.1
(M+H).sup.+.
[1030] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.65 (quart, 1H,
J=4.6 Hz); 8.47 (s, 1H); 8.38 (bs, 1H); 8.24 (s, 1H); 7.21 (dd, 1H,
J=8.7 Hz, J=2.7 Hz); 7.06 (bs, 1H); 6.99 (d, 1H, J=8.7 Hz); 6.31
(dd, 1H, J=5.4 Hz, J=2.1 Hz); 5.99-5.97 (mult, 1H); 4.97-4.95
(mult, 1H); 4.65 (bs, 2H); 4.50-4.45 (mult, 1H); 4.35-4.29 (mult,
1H); 4.01-3.88 (mult, 2H); 3.83-3.72 (mult, 2H); 3.65-3.60 (mult,
1H); 3.59-3.53 (mult, 1H); 2.70-2.60 (mult, 4H); 2.05-1.95 (mult,
1H); 1.75-1.62 (mult, 1H).
Preparation E10
[1031]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(4-methyl-benzyloxy)-benzylam-
ino]-purin-yl)-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1032] Mp 88.0-95.0.degree. C.
[1033] C.sub.26H.sub.26ClN.sub.9O.sub.4. MW 564.01. MS 564.2
(M+H).sup.+.
[1034] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.66 (quart, 1H,
J=4.8 Hz); 8.47 (s, 1H); 8.42 (bs, 1H); 8.22 (s, 1H); 7.45 (d, 1H,
J=7.1 Hz); 7.27-7.16 (mult, 4H); 7.14 (d, 1H, J=8.7 Hz); 7.06 (bs,
1H); 6.32 (d, 1H, J=5.4 Hz); 5.99 (d, 1H, J=6.4 Hz); 5.15 (s, 2H);
4.96 (quart, 1H, J=5.5 Hz); 4.68 (bs, 2H); 4.50-4.45 (mult, 1H);
4.32 (d, 1H, J=2.9 Hz); 2.67 (d, 3H, J=4.8 Hz); 2.34 (s, 3H).
Preparation E11
[1035]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(2-methyl-benzyloxy)-benzylam-
ino]-purin-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1036] Mp 58.0-62.0.degree. C.
[1037] C.sub.26H.sub.26ClN.sub.9O.sub.4. MW 564.01. MS 564.2
(M+H).sup.+.
[1038] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.66 (quart, 1H,
J=4.8 Hz); 8.48 (s, 1H); 8.41 (bs, 1H); 8.24 (s, 1H); 7.35 (d, 1H,
J=7.9 Hz); 7.21 (d, 1H, J=8.5 Hz); 7.20-7.15 (mult, 3H); 7.06 (d,
1H, J=8.7 Hz); 6.99 (d, 1H, J=8.7 Hz); 6.32 (d, 1H, J=5.4 Hz); 5.99
(d, 1H, J=6.4 Hz); 5.12 (s, 2H); (quart, 1H, J=5.8 Hz); 4.68 (bs,
2H) 4.50-4.47 (mult, 1H); 4.32 (d, 1H, J=3.1 Hz); 2.67 (d, 3H,
J=4.8 Hz); 2.28 (s, 3H).
Preparation E12
[1039]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(3-methyl-benzyloxy)-benzylam-
ino]-purin-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1040] Mp 80.0-84.0.degree. C.
[1041] C.sub.26H.sub.26ClN.sub.9O.sub.4. MW 564.01. MS 564.2
(M+H).sup.+.
[1042] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.66 (quart, 1H,
J=4.8 Hz); 8.48 (s,1H); 8.41 (bs, 1H); 8.24 (s, 1H); 7.28-7.24
(mult, 3H); 7.22 (dd, 1H, J=8.7 Hz, J=2.5 Hz); 7.13-7.05 (mutt,
3H); 6.31 (d, 1H, J=5.4 Hz); 5.99 (d, 1H, J=6.4 Hz); 5.13 (s, 2H);
4.96 (quart, 1H, J=5.7 Hz); 4.69 (bs, 2H); 4.50-4.45 (mult, 1H);
4.32 (d, 1H, J=3.1 Hz); 2.67 (d, 3H, J=4.8 Hz); 2.28 (s, 3H).
Preparation E13
[1043]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(2-methoxybenzyloxy)benzylami-
no]-purin-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1044] C.sub.26H.sub.26ClN.sub.9O.sub.5. MW 580.01. MS 580.1
(M+H).sup.+.
[1045] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.63 (quart, 1H,
J=4.8 Hz); 8.45 (s, 1H); 8.36 (bs, 1H); 8.21 (s, 1H); 7.41 (dd, 1H,
J=7.7 Hz, J=1.7 Hz); (t, 1H, J=7.2 Hz); 7.19 (dd, 1H, J=8.7 Hz,
J=2.7 Hz); 7.05-6.99 (mutt, 3H); 6.92 (t, 1H, J=7.5 Hz); 6.28 (d,
1H, J=5.4 Hz); 5.96 (d, 1H, J=6.4 Hz); 5.09 (s, 2H); 4.98-4.90
(mult, 1H); 4.65 (bs, 2H); 4.48-4.42 (mult, 1H); 4.29 (d, 1H, J=3.1
Hz); 3.80 (s, 3H); 2.64 (d, 3H, J=4.8 Hz).
Preparation E14
[1046]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(furan-3-ylmethoxy)benzylamin-
o]-purin-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1047] Mp 86.0-90.0.degree. C.
[1048] C.sub.23H.sub.22ClN.sub.9O.sub.5. MW 539.94. MS 540.1
(M+H).sup.+.
[1049] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.66 (quart, 1H,
J=4.6 Hz); 8.48 (s, 1H); 8.39 (bs, 1H); 8.24 (s, 1H); 7.80 (s, 1H);
7.66 (s, 1H); 7.22 (dd, 1H, J=8.7 Hz, J=2.5 Hz); 7.11 (d, 1H, J=8.7
Hz); 7.06 (bs, 1H); 6.60 (s, 1H); 6.31 (d, 1H, J=5.4 Hz); 5.99 (d,
1H, J=6.4 Hz); 5.03 (s, 2H); 4.96 (quart, 1H, J=5.6 Hz); 4.64 (bs,
2H); 4.49-4.47 (mult, 1H); 4.32 (d, 1H, J=3.1 Hz); 2.67 (d, 3H,
J=4.6 Hz).
Preparation E15
[1050]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(4-methoxy-benzyloxy)-benzyla-
mino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide
[1051] C.sub.26H.sub.26ClN.sub.9O.sub.5. MW 580.01. MS 580.0
(M+H).sup.+.
[1052] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.63 (quart, 1H,
J=4.6 Hz); 8.45 (s, 1H); 8.37 (bs, 1H); 8.21 (s, 1H); 7.37 (d, 2H,
J=8.5 Hz); 7.19 (dd, 1H, J=8.5 Hz, J=2.5 Hz); 7.05 (d, 2H, J=8.9
Hz); 6.89 (d, 2H, J=8.7 Hz); 6.28 (d, 1H, J=4.8 Hz); 5.96 (d, 1H,
J=6.2 Hz); 5.06 (s, 2H); 4.98-4.90 (mult, 1H); 4.63 (bs, 2H);
4.46-4.42 (mult, 1H); 4.29 (d, 1H, J=2.9 Hz); 3.70 (s, 3H); 2.64
(d, 3H, J=4.6 Hz).
Preparation E16
[1053]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-cyclopentylmethoxybenzylamino-
)-purin-9-yl]4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1054] C.sub.24H.sub.28ClN.sub.9O.sub.4. MW 542.00. MS 541.8
(M+H).sup.+.
[1055] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.64 (quart, 1H,
J=4.8 Hz); 8.45 (s, 1H); 8.33 (bs, 1H); 8.21 (s, 1H); 7.17 (dd, 1H,
J=8.7 Hz, J=2.9 Hz); 7.00 (bs, 1H), 6.95 (d, 1H, J=8.7 Hz); 6.29
(d, 1H, J=5.6 Hz); 5.96 (d, 1H, J=6.2 Hz); 4.95-4.90 (mult, 1H);
4.62 (bs, 2H); 4.47-4.43 (mult, 1H); 4.29 (d, 1H, J=3.1 Hz); 3.85
(d, 2H, J=6.9 Hz); 2.64 (d, 3H, J=4.8 Hz); 2.32-2.21 (mult, 1H);
1.78-1.66 (mult, 2H); 1.60-1.42 (mult, 4H); 1.37-1.25 (mult,
2H).
Preparation E17
[1056]
(2S,3S,4R,5R)3-Azido-5-(6-{5-chloro-2-[3-(2-morpholin4-yl-ethoxy]-b-
enzylamino}-purin-9-yl)4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1057] C.sub.31H.sub.35ClN.sub.10O.sub.6. MW 679.14. MS 678.7
(M+H).sup.+.
[1058] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.62 (quart, 1H,
J=5.0 Hz); 8.45 (s, 1H); 8.40 (bs, 1H); 8.22 (s, 1H); 7.27-7.17
(mult, 2H); 7.05-7.03 (mult, 1H); 7.03-6.98 (mult, 3H); 6.84 (d,
1H, J=8.3 Hz); 6.30-6.25 (mult, 1H); 5.99-5.92 (mult, 1H); 5.12 (s,
2H); 4.95-4.88 (mult, 1H); 4.73-4.65 (mult, 2H); 4.45-4.41 (mult,
1H); 4.30-4.26 (mult, 1H); 4.07-4.00 (mult, 2H); 3.55-3.45 (mult,
4H); 2.67-2.60 (mult, 5H); 2.42-2.37 (mult, 4H).
Preparation E18
[1059]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(2,2,7,7-tetramethyltetrahydr-
o-bis[1,3]dioxolo[4,5-b;4',5'-d]pyran-5-ylmethoxy)benzylamino]purin-9-yl}4-
-hydroxytetrahydrofuran-2-carboxylic acid methylamide
[1060] C.sub.30H.sub.36ClN.sub.9O.sub.9. MW 702.13. MS 702.1
(M+H).sup.+.
[1061] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.72-8.65 (mult,
1H); 8.47 (s, 1H); 8.38 (bs, 1H); 8.25 (s, 1H); 7.21 (dd, 1H, J=8.5
Hz, J=2.3 Hz); 7.06 (bs, 1H); 7.03 (d, 1H, J=8.5 Hz); 6.38 (bs,
1H); 5.98 (d, 1H, J=6.4 Hz); 5.48 (d, 1H, J=4.8 Hz); 4.95 (t, 1H,
J=5.8 Hz); 4.65 (bs, 2H); 4.60 (d, 1H, J=7.9 Hz); 4.51-4.45 (mult,
1H); 4.39-4.35 (mult, 1H); 4.35-4.29 (mult, 2H); 4.21-4.16 (mult,
1H); 4.08-3.99 (mult, 2H); 2.67 (d, 3H, J=4.6 Hz); 1.35 (d, 6H,
J=11.4 Hz); 1.25 (s, 6H).
Preparation F1
[1062]
(5-{6-[5-Chloro-2-(3-methylisoxazol-5-ylmethoxy)benzylamino]purin-9-
-yl}4-hydroxy-2-methylcarbamoyltetrahydrofuran-3-yl)-carbamic acid
tert-butyl ester
[1063] To a mixture of
[5-(6-chloropurin-9-yl)-4-hydroxy-2-methylcarbamoyl-
-tetrahydrofuran-3-yl]-carbamic acid tert-butyl ester (500 mg, 1.2
mmol) and (3-methylisoxazol-5-ylmethoxy)benzyl amine (1.4 mmol) in
anhydrous ethanol (10 mL) was added triethylamine (0.5 mL, 3.6
mmol) and the reaction was heated to 65.degree. C.
[1064] The reaction was stirred at reflux for 15 hours under
anhydrous conditions. The solvent was then removed by rotary
evaporation and the product was preadsorbed onto silica gel and
purified by flash chromatography to afford the title compound.
[1065] Mp 135.0-137.0.degree. C.
[1066] C.sub.28H.sub.33ClN.sub.8O.sub.7. MW 629.08. MS 629.2
(M+H).sup.+.
[1067] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.55 (bs, 1H);
8.41-8.33 (mult, 2H); 8.19 (s, 1H); 7.23 (dd, 1H, J=8.7 Hz, J=2.7
Hz); 7.12 (d, 1H, J=8.7 Hz); 7.07 (bs, 1H); 6.96 (d, 1H, J=6.0 Hz);
6.47 (s, 1H); 5.98 (d, 1H, J=4.6 Hz); 5.91 (bs, 1H); 5.29 (s, 2H);
4.63 (bs, 2H); 4.55-4.43 (mult, 1H); 4.28-4.17 (mult, 2H); 2.63 (d,
3H, J=4.6 Hz); 2.20 (s, 3H); 1.35 (s, 9H).
Preparation G1
[1068]
(2R,3R,4S,5S)(2-{[9-(4-Azido-3-hydroxy-5-methylcarbamoyl-tetrahydro-
-furan-2-yl)-9H-purin-6-ylamino]-methyl}-4-chloro-phenoxy)-acetic
acid tert-butyl ester
[1069] Triethyl amine (0.3 mL, 2.1 mmol) was added to a solution of
(2S,3S,4R,5R)3-azido-5-(6-chloro-purin-9-yl)-4-hydroxy-tetrahydro-furan-2-
-carboxylic acid methylamide (302 mg, 0.9 mmol) and
2-aminomethyl-4-chloro-phenoxy acetic acid tert-butyl ester (290
mg, 1.07 mmol) in ethanol at ambient temperature. The reaction was
heated to 70.degree. C. for 2.5 hours, cooled, and the resultant
solid was washed with cold ethanol, and dried to afford 420 mg
(81%) of the title compound as a colorless solid. Alternatively,
the product could be purified by flash chromatography (2-5%
methanol/dichloromethane). MS 574 (M+H).sup.+. .sup.1H NMR (400
MHz, CD.sub.3OD) .delta.8.28 (bs, 1H); 8.24 (s, 1H); 7.32 (bs, 1H);
7.18 (dd, 1H, J=8.9, 2.6 Hz); 6.92 (d, 1H, J=8.9 Hz); 5.95 (d, 1H,
J=7.1 Hz); 5.03 (dd, 1H, J=7.1, 4.9 Hz); 4.82 (bs, 2H); 4.83 (s,
2H); 4.39 (m, 2H); 3.0 (s, 1H); 2.8 (s, 3H); 1.4 (s, 9H).
[1070] In an analogous manner the following compounds, Preparations
G2-G30, were prepared from the appropriate starting material using
analogous procedures to Preparation G1.
Preparation G2
[1071]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(3-methylisoxazol-5-ylmethoxy-
) benzylamino]purin-9-yl}4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1072] Mp 94.0-98.0.degree. C.; C.sub.23H.sub.23ClN.sub.10O.sub.5.
MW 554.96. MS 555.1 (M+H).sup.+.
[1073] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.8.63 (quart, 1H,
J=4.6 Hz); 8.46 (s, 1H);,8.41 (bs, 1H); 8.21 (s, 1H); 7.23 (dd, 1H,
J=8.7 Hz, J=2.5 Hz); 7.11 (d, 1H, J=8.7 Hz); 7.06 (bs, 1H); 6.48
(s, 1H); 6.29 (d, 1H, J=5.6 Hz); 5.97 (d, 1H, J=6.4 H (s, 2H); 4.94
(quart, 1H, J=5.6 Hz); 4.63 (bs, 2H); 4.48-4.42 (mult, 1H); 4.30
(d, 1H, J=2.7 Hz); 2.64 (d, 3H, J=4.6 Hz); 2.19 (s, 3H).
Preparation G3
[1074]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(tetrahydrofuran-3-ylmethoxy)-
-benzylamino]-purin-9-yl)4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1075] C.sub.23H.sub.26ClN.sub.9O.sub.5. MW 543.97. MS 544.1
(M+H).sup.+.
[1076] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.65 (quart, 1H,
J=4.6 Hz); 8.47 (s, 1H); 8.38 (bs, 1H); 8.24 (s, 1H); 7.21 (dd, 1H,
J=8.7 Hz, J=2.7 Hz); 7.06 (bs, 1H); 6.99 (d, 1H, J=8.7 Hz); 6.31
(dd, 1H, J.=5.4 Hz, J=2.1 Hz); 5.99-5.97 (mult, 1H); 4.97-4.95
(mult, 1H); 4.65 (bs, 2H); 4.50-4.45 (mult, 1H); 4.35-4.29 (mult,
1H); 4.01-3.88 (mult, 2H); 3.83-3.72 (mult, 2H); 3.65-3.60 (mult,
1H); 3.59-3.53 (mult, 1H); 2.70-2.60 (mult, 4H); 2.05-1.95 (mult,
1H); 1.75-1.62 (mult, 1H).
Preparation G4
[1077]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(tetrahydrofuran-3-ylmethoxy)-
-benzylamino]-purin-9-yl}4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1078] C.sub.23H.sub.26ClN.sub.9O.sub.5. MW 543.97. MS 544.1
(M+H).sup.+.
[1079] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.8.65 (quart, 1H,
J=4.6 Hz); 8.47 (s, 1H); 8.38 (bs, 1H); 8.24 (s, 1H); 7.21 (dd, 1H,
J=8.7 Hz, J=2.7 Hz); 7.06 (bs, 1H); 6.99 (d, 1H, J=8.7 Hz); 6.31
(dd, 1H, J=5.4 Hz, J=2.1 Hz); 5.99-5.97 (mult, 1H); 4.97-4.95
(mult, 1H); 4.65 (bs, 2H); 4.50-4.45 (mult, 1H); 4.35-4.29 (mult,
1H); 4.01-3.88 (mult, 2H); 3.83-3.72 (mult, 2H); 3.65-3.60 (mult,
1H); 3.59-3.53 (mult, 1H); 2.70-2.60 (mult, 4H); 2.05-1.95 (mult,
1H); 1.75-1.62 (mult, 1H).
Preparation G5
[1080]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(furan-2-ylmethoxy)-benzylami-
no]-purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1081] Mp 182.0-185.0.degree. C.
[1082] C.sub.23H.sub.22ClN.sub.9O.sub.5. MW 539.94. MS 540.1
(M+H).sup.+.
[1083] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.8.66 (d, 1H,
J=4.6 Hz); 8.47 (s, 1H); 8.38 (bs, 1H); 8.23 (s, 1H); 7.68 (d, 1H,
J=1.9 Hz); 7.23 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.17 (d, 1H, J=8.7
Hz); 7.03 (bs, 1H); 6.60 (d, 1H, J=3.1 Hz); 6.45 (dd, 1H, J=3.1 Hz,
J=1.9 Hz); 6.31 (d, 1H, J=5.4 Hz); 5.98 (d, 1H, J=6.2 Hz); 5.13 (s,
2H); 4.96 (quart, 1H, J=6.0 Hz); 4.60 (bs, 2H); 4.51-4.45 (mult,
1H); 4.31 (d, 1H, J=3.1 Hz); 2.66 (d, 3H, J=4.6 Hz).
Preparation G6
[1084]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(2,5-dimethylfuran-3-ylmethox-
y)benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic
acid methylamide
[1085] Mp 85.0-87.0.degree. C.
[1086] C.sub.25H.sub.26ClN.sub.9O.sub.5. MW 568.00. MS 568.1
(M+H).sup.+.
[1087] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta.8.63 (quart, 1H,
J=4.6 Hz); 8.45 (s, 1H); 8.33 (bs, 1H); 8.20 (s, 1H); 7.19 (dd, 1H,
J=8.7 Hz, J=2.7 Hz); 7.05 (d, 1H, J=8.7 Hz); 7.02 (bs, 1H); 6.28
(d, 1H, J=5.4 Hz); 6.06 (s, 1H); 5.96 (d, 1H, J=6.4 Hz); 4.93
(quart, 1H, J=5.2 Hz); 4.86 (s, 2H); 4.59 (bs, 2H); 4.47-4.42
(mult, 1H); 4.29 (d, 1H, J=3.1 Hz); 2.64 (d, 3H, J=4.6 Hz); 2.21
(s, 3H); 2.14 (s, 3H).
Preparation G7
[1088]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(5-dimethylaminomethylfuran-2-
-ylmethoxy)benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic
acid methylamide
[1089] C.sub.26H.sub.29ClN .sub.10O.sub.5. MW 597.04. MS 597.1
(M+H).sup.+.
[1090] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.63 (quart, 1H,
J=4.6 Hz); 8.45 (s, 1H); 8.34 (bs, 1H); 8.20 (s, 1H); 7.22-7.16
(mult, 1H); 7.13 (d, 1H, J=8.7 Hz); 7.00 (bs, 1H); 6.49 (d, 1H,
J=2.7 Hz); 6.29 (bs, 1H); 6.22 (d, 1H, J=3.1 Hz); 5.96 (d, 1H,
J=6.0 Hz); 5.06 (s, 2H); 4.92 (bs, 1H); 4.56 (bs, 2H); 4.48-4.41
(mult, 1H); 4.36-4.22 (mult, 1H); 3.28 (bs, 2H); 2.63 (d, 1H, J=4.6
Hz); 2.14-2.04 (mult, 6H).
Preparation G8
[1091]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(thiazol-2-ylmethoxy)benzylam-
ino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1092] Mp 88.0-92.0.degree. C.
[1093] C.sub.22H.sub.21ClN.sub.10O.sub.4S. MW 556.99. MS 557.1
(M+H).sup.+.
[1094] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.66 (quart, 1H,
J=4.4 Hz); 8.52-8.40 (mult, 2H); 8.24 (s, 1H); 7.84 (d, 1H, J=2.9
Hz); 7.77 (d, 1H, J=2.9 Hz); 7.25 (d, 1H, J=8.5 Hz); 7.15 (d, 1H,
J=8.5 Hz); 7.08 (bs, 1H); 6.31 (d, 1H, J=5.4 Hz); 5.98 (d, 1H,
J=6.4 Hz); 5.51 (s, 2H); 5.00-4.91 (mult, 1H); 4.72 (bs, 2H); 4.49
(bs, 1H); 4.32 (bs, 1H); 2.66 (d, 3H, J=4.4 Hz).
Preparation G9
[1095]
(2S,3S,4R,5R)3-Azido-5-{6-[2-(benzothiazol-2-ylmethoxy)-5-chloroben-
zylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1096] Mp 95.0-100.0.degree. C.
[1097] C.sub.26H.sub.23ClN.sub.10O.sub.4S. MW 607.05. MS 607.0
(M+H).sup.+.
[1098] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.66 (quart, 1H,
J=4.4 Hz); 8.49 (bs, 2H); 8.24 (s, 1H); 8.11 (d, 1H, J=8.1 Hz);
8.01 (d, 1H, J=8.1 Hz); 7.52 (t, 1H, J=7.7 Hz); 7.44 (t, 1H, J=7.7
Hz); 7.26 (d, 1H, J=8.3 Hz); 7.17 (d, 1H, J=8.3 Hz); 7.10 (bs, 1H);
6.32 (d, 1H, J=5.2 Hz); 5.99 (d, 1H, J=6.4 Hz); 5.67 (s, 2H);
5.00-4.96 (mult, 1H); 4.80 (bs, 2H); 4.49 (bs, 1H); 4.32 (bs, 1H);
2.66 (d, 3H, J=4.4 Hz).
Preparation G10
[1099]
(2S,3S,4R,5R)3-Azido-5-{6-[2-(benzofuran-2-ylmethoxy)-5-chlorobenzy-
lamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1100] Mp 89.0-100.0.degree. C.
[1101] C.sub.27H.sub.24ClN.sub.9O.sub.5. MW 590.00. MS 590.1
(M+H).sup.+.
[1102] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.65 (quart, 1H,
J=4.4 Hz); 8.47 (s, 1H); 8.41 (bs, 1H); 8.20 (s, 1H); 7.63 (d, 1H,
J=7.9 Hz); 7.57 (d, 1H, J=7.9 Hz); 7.31 (t, 1H, J=7.5 Hz);
7.26-7.20 (mult, 3H); 7.07 (mult, 2H); 6.31 (d, 1H, J=5.2 Hz); 5.98
(d, 1H, J=6.4 Hz); 5.35 (s, 2H); 4.95 (quart, 1H, J=5.8 Hz); 4.66
(bs, 2H); 4.48 (bs, 1H
Preparation G11
[1103]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(isothiazol-5-ylmethoxy)benzy-
lamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1104] Mp 96.0-99.0.degree. C.
[1105] C.sub.22H.sub.21ClN.sub.10O.sub.4S. MW 556.99. MS 557.1
(M+H).sup.+.
[1106] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.63 (quart, 1H,
J=4.6 Hz); 8.51 (s, 1H); 8.46 (s, 1H); 8.42 (bs, 1H); 8.22 (s, 1H);
7.45 (s, 1H); 7.24 (dd, 1H, J=8.7 Hz, J=2.5 Hz); 7.12-7.07 (mult,
2H); 6.29 (d, 1H, J=5.4 Hz); 5.97 (d, 1H, J=6.4 Hz); 5.57 (s, 2H);
4.94 (quart, 1H, J=5.8 Hz); 4.67 (bs, 2H); 4.49-4.41 (mult, 1H);
4.30 (d, 1H, J=3.1 Hz); 2.65 (d, 3H, J=4.6 Hz).
Preparation G12
[1107]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(thiophen-2-ylmethoxy)benzyla-
mino]purin-9-yl}4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1108] Mp91.0-112.0.degree. C.
[1109] C.sub.23H.sub.22ClN.sub.9O.sub.4S. MW 556.01. MS 556.1
(M+H).sup.+.
[1110] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.63 (quart, 1H,
J=4.6 Hz); 8.46 (s, 1H); 8.42 (bs, 1H); 8.22 (s, 1H); 7.52 (d, 1H,
J=5.0 Hz); 7.25-7.18 (mult, 2H); 7.13 (d, 1H, J=8.9 Hz); 7.03 (bs,
1H); 7.01-6.98 (mult, 1H); 6.30 (bs, 1H); 5.97 (d, 1H, J=6.2 Hz);
5.34 (s, 2H); 4.97-4.90 (mult, 1H); 4.62 (bs, 2H); 4.48-4.41 (mult,
1H); 4.30 (d, 1H, J=3.1 Hz); 2.64 (d, 3H, J=4.6 Hz).
Preparation G13
[1111]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(quinolin-2-ylmethoxy)benzyla-
mino]purin-9-yl)4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1112] Mp 117.0-120.0.degree. C.
[1113] C.sub.28H.sub.25ClN.sub.10O.sub.4. MW 601.03. MS 601.1
(M+H).sup.+.
[1114] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.64 (quart, 1H,
J=4.6 Hz); 8.47 (bs, 1H), 8.46 (s, 1H); 8.37 (d, 1H, J=8.5 Hz);
8.24 (s, 1H); 8.03-7.92 (mult, 2H); 7.80-7.67 mult, 2H); 7.58 (t,
1H, J=7.1 Hz); 7.18 (dd, 1H, J=8.5 Hz, J=2.1 Hz); 7.12 (bs, 1H);
7.07 (d, 1H, J=8.5 Hz); 6.30 (d, 1H, J=5.4 Hz); 5.97 (d, 1H, J=6.4
Hz); 5.42 (s, 2H); 4.95 (quart, 1H, J=5.2 Hz); 4.78 (bs, 2H);
4.51-4.44 (mult, 1H); 4.30 (d, 1H, J=3.1 Hz); 2.64 (d, 3H, J=4.6
Hz).
Preparation G14
[1115]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(4-methyl-[1,2,3]thiadiazol-5-
-ylmethoxy)benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic
acid methylamide
[1116] Mp 105.0-107.0.degree. C.
[1117] C.sub.22H.sub.22ClN.sub.11O.sub.4S. MW 572.01. MS 572.0
(M+H).sup.+.
[1118] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.63 (quart, 1H,
J=4.8 Hz); 8.46 (s, 1H); 8.42 (bs, 1H); 8.20 (s, 1H); 7.27 (dd, 1H,
J=8.7 Hz, J=2.7 Hz); 7.16 (d, 1H, J=8.7 Hz); 7.09 (bs, 1H); 6.30
(d, 1H, J=5.4 Hz); 5.96 (d, 1H, J=6.6 Hz); 5.57 (s, 2H); 4.94
(quart, 1H, J=5.2 Hz); 4.63 (bs, 2H); 4.49-4.42 (mult, 1H); 4.30
(d, 1H, J=5.4 Hz); 2.70-2.60 (mult, 6H).
Preparation G15
[1119]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(naphthalen-1-ylmethoxy)
benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1120] Mp 115.0-119.0.degree. C.
[1121] C.sub.29H.sub.26ClN.sub.9O.sub.4. MW 600.04. MS 600.1
(M+H).sup.+.
[1122] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.63 (quart, 1H,
J=4.8 Hz); 8.44 (s, 1H); 8.38 (bs, 1H); 8.19 (s, 1H); 8.14 (d, 1H,
J=7.9 Hz); 7.98-7.86 (mult, 2H); 7.70 (d, 1H, J=6.6 Hz); 7.59-7.43
(mult, 3H); 7.32-7.21 (mult, 2H); 7.06 (bs, 1H); 6.29 (d, 1H, J=5.2
Hz); 5.96 (d, 1H, J=6.4 Hz); 5.61 (s, 2H); 4.93 (quart, 1H, J=5.6
Hz); 4.62 (bs, 2H); 4.49-4.42 (mult, 1H); 4.30 (d, 1H, J=2.9 Hz);
2.64 (d, 3H, J=4.8 Hz).
Preparation G16
[1123]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-hydroxybenzylamino)-purin-9-y-
l]-4-hydroxytetrahydrofuran-2-carboxylic acid methylamide
[1124] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 10.04 (bs, 1H);
8.63 (quart, 1H, J=4.6 Hz); 8.45 (s, 1H); 8.40 (bs, 1H); 8.25 (s,
1H); 7.05 (dd, 1H, J=8.5 Hz, J=2.5 Hz); 7.00 (bs, 1H); 6.77 (d, 1H,
J=8.5 Hz); 6.30 (d, 1H, J=5.4 Hz); 5.96 (d, 1H, J=6.2 Hz); 4.94
(quart, 1H, J=5.8 Hz); 4.56 (bs, 2H); 4.49-4.43 (mult, 1H); 4.30
(d, 1H, J=2.9 Hz); 2.65 (d, 3H, J=4.6 Hz).
Preparation G17
[1125]
(2S,3S,4R,5R)3-Azido-5-[6-(2-benzyloxy-5-chloro-benzylamino)-2-chlo-
ro-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide.
Preparation G18
[1126]
(2S,3S,4R,5R)3-Azido-5-[6-(5-chloro-2-phenethyloxy-benzylamino)-pur-
in-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide
Preparation G19
[1127]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(1-phenyl-ethoxy)-benzylamino-
]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide
Preparation G20
[1128]
(2S,3S,4R,5R)3-Azido-5-{6-[5-chloro-2-(4,5-dihydro-1H-imidazol-2-yl-
methoxy)-benzylamino]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic
acid methylamide
Preparation G21
[1129]
(2S,3S,4R,5R)3-Azido-5-{6-[1-(2-benzyloxy-5-chloro-phenyl)-ethylami-
no]-purin-9-yl}-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide
Preparation G22
[1130]
(2S,3S,4R,5R)3-Azido-5-[6-(2-benzyloxy-5-bromo-benzylamino)-purin-9-
-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide
Preparation G23
[1131]
(2S,3S,4R,5R)3-Azido-5-[6-(2-benzyloxy-5-fluoro-benzylamino)-purin--
9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide
Preparation G24
[1132]
(2S,3S,4R,5R)3-Azido-5-[6-(2-benzyloxy-5-iodo-benzylamino)-purin-9--
yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide
Preparation G25
[1133]
(2S,3S,4R,5R)3-Azido-5-[6-(2-benzyloxy-5-trifluoromethyl-benzylamin-
o)-purin-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid
methylamide
Preparation G26
[1134]
(2S,3S,4R,5R)3-Azido-5-[6-(2-benzyloxy-5-cyano-benzylamino)-purin-9-
-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide
Preparation G27
[1135]
(2S,3S,4R,5R)3-Azido-5-[6-(2-benzyloxy-5-methyl-benzylamino)-purin--
9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide
Preparation G28
[1136]
(2S,3S,4R,5R)3-Azido-5-[6-(2-benzyloxy-5-vinyl-benzylamino)-purin-9-
-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide
Preparation G29
[1137]
(2S,3S,4R,5R)3-Azido-5-[6-(2-benzyloxy-5-ethynyl-benzylamino)-purin-
-9-yl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid methylamide
Preparation G30
[1138] 3-Azido-5-{6-[5-chloro-2-(3,5-dimethylisoxazol-4-ylmethoxy)
benzylamino]purin-9-yl}-4-hydroxytetrahydrofuran-2-carboxylic acid
methylamide
[1139] C.sub.24H.sub.25ClN.sub.10O.sub.5. MW 568.98. MS 569.1
(M+H).sup.+.
[1140] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 5 8.69 (quart,
1H, J=4.8 Hz); 8.40 (s, 1H); 8.35 (bs, 1H); 8.14 (s, 1H); 7.25 (dd,
1H, J=8.7 Hz, J=2.7 Hz); 7.11 (d, 1H, J=8.7 Hz); 7.07 (bs, 1H);
6.28 (d, 1H, J=5.6 Hz); 6.05 (d, 1H, J=6.6 Hz); 4.95 (s, 2H);
4.85-4.77 (mult, 1H); 4.67-4.52 (mult, 2H); 4.49-4.42 (mult, 1H);
4.39-4.32 (mult, 1H); 2.65 (d, 3H, J=4.8 Hz); 2.38 (s, 3H); 2.22
(s, 3H).
Preparation H1
[1141]
(2S,3S,4R,5R)3-Azido-5-(6-chloro-purin-9-yl)4-hydroxy-tetrahydro-fu-
ran-2-carboxylic acid methylamide
[1142] Triethylamine (4.4 mL, 0.032 mL) was added to a solution of
(2S,3S,4R,5R)3-azido-5-(6-chloro-purin-9-yl)-4-acetoxy-tetrahydro-furan-2-
-carboxylic acid methylamide (4 g, 0.011 mmol) in methanol (80 mL).
After stirring for 15 hours, the mixture was concentrated and the
residue purified by flash chromatography (5%
methanol/dichloromethane) to afford 2.7 g (77%) of the title
compound as a colorless solid. MS 339 (M+H).sup.+.sup.1H NMR (400
MHz, CDCl.sub.3) .delta. 8.72 (s, 1H); 8.2 (s, 1H); 7.62 (bs, 1H);
5.98 (d, 1H, J=6.7 Hz); 5.07 (t, 1H, J=6.2 Hz); 4.65 (dd, 1H,
J=5.4, 2.7 Hz); 4.58 (m, 1H); 4.2 (bs, 1H); 2.88 (d, 1H, J=4.9 Hz);
1.67 (bs, 1H).
Preparation I1
[1143] Alternate preparation of
(2S,3S,4R,5R)3-Azido-5-(6-chloropurin-9-yl-
)4-hydroxytetrahydrofuran-2-carboxylic acid methylamide
[1144] To a solution of acetic acid
4-azido-2-(6-chloropurin-9-yl)-5-methy-
lcarbamoyl-tetrahydrofuran-3-yl ester (1.1 g, 2.9 mmol) in
anhydrous methanol (100 mL), cooled to 0.degree. C., was added
methylamine (1.2 mL, 8.6 mmol). The solution was stirred for 2 h at
room temperature, under anhydrous conditions. After removal of
solvent by rotary evaporation, the resulting solid was purified via
flash chromatography (7% MeOH/CH.sub.2Cl.sub.2) to yield the title
compound as a white foam.
[1145] C.sub.11H.sub.11ClN.sub.8O.sub.3. MW 338.72. MS 339.1
(M+H).sup.+.
[1146] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.01 (s, 1H);
8.81 (s, 1H); 8.22 (quart, 1H, J=4.2 Hz); 6.41 (dd, 1H, J=5.2 Hz,
J=2.1 Hz); 6.12 (d, 1H, J=5.2 Hz); 5.03 (quart, 1H, J=5.2 Hz);
4.57-4.47 (mult, 1H); 4.41 (d, 1H, J=3.9 Hz); 2.61 (d, 3H, J=4.2
Hz).
Preparation J1
Glycosidation
[1147]
(2S,3S,4R,5R)3-Azido-5-(6-chloro-purin-9-yl34-acetoxy-tetrahydro-fu-
ran-2-carboxylic acid methylamide
[1148] 6-Chloropurine (20.4 g, 0.132 mol) was suspended in
hexamethyl disilazane (165 mL) and heated at 110.degree. C. After 2
h, the now homogeneous mixture was concentrated and the solid
residue reconcentrated from toluene 2.times. and placed under high
vac for 1 hour. The resulting solid was combined with
1,2-bis-O-acetyl-3-azido-3-deoxy-D-ribofuranuroni- c acid methyl
amide (12.7 g, 0.044 mol) and dissolved in anhydrous dichloroethane
(350 mL). Powdered 4A molecular sieves (15 g) were added and the
mixture stirred for 15 minutes. TMSOTf (16.0 mL, 0.088 mol) was
added and the reaction was then heated to 60.degree. C. for two
hours, cooled and quenched by the careful addition of saturated
sodium bicarbonate solution (200 mL). Ethyl acetate (350 mL) was
added and the mixture was filtered through sintered glass. The
filtrate was extracted with ethyl acetate (3.times.) and the
combined organic layers were washed with brine, dried
(Na.sub.2SO.sub.4), filtered and concentrated.
[1149] The residue was purified by flash chromatography (15% then
20% acetoneldichloromethane) to afford the title compound (10.6 g)
as an off-white foam. MS 381 (M+H).sup.+. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.8 (s, 1H); 8.2 (s, 1H); 7.6 (bs, 1H); 6.13
(d, 1H, J=7.0 Hz); 5.87 (dd, 1H, J=7.0, 5.6 Hz); 4.85 (dd, 1H,
J=5.6, 3.0 Hz); 4.58 (d, 1H, J=3.0 Hz); 2.9 (d, 1H, J=4.9 Hz); 2.11
(s, 3H).
[1150] The following compound, Preparation J2, was prepared from
the appropriate starting material using analogous procedures to
Preparation J1.
Preparation J2
[1151]
(2S,3S,4R,5R)3-Azido-5-(2,6-dichloro-purin-9-yl)4-hydroxy-tetrahydr-
o-furan-2-carboxylic acid methylamide
Preparation K1
[1152] 1,2-bis-O-acetyl-3-azido-3-deoxy-D-ribofuranuronic acid
methyl amide
3-Azido-3-deoxy-1,2-0-isopropylidene-.alpha.-D-ribofuranuronic acid
methyl amide (12 g, 0.05 mmol) was dissolved in acetic acid (150
mL) and acetic anhydride (50 mL).
[1153] The mixture was cooled in an ice bath and a solution of
sulfuric acid (1 mL dissolved in 5 mL acetic acid) was added. The
mixture was allowed to warm to room temperature and stirred for 18
hours. The reaction mixture was added dropwise to a saturated
solution of sodium bicarbonate (2 L), and then extracted with
chloroform (3.times.). The combined organic layers were washed with
water (2.times.) and sat. NaHCO.sub.3 (2.times.) and brine
(1.times.), dried (Na.sub.2SO.sub.4) filtered and concentrated to
afford a mixture of anomeric acetates as a tan oil.
Preparation L1
[1154]
3-Azido-3-deoxy-1,2-O-isopropylidene-.alpha.-D-ribofuranuronic acid
methyl amide Oxalyl chloride (15 mL) was added to a solution of
3-azido-3-deoxy-.alpha.-D-ribofuranuronic acid (30 g) in anhydrous
THF (250 mL) at 0.degree. C. DMF (1 mL) was added and the reaction
was allowed to warm to room temperature at which time gas evolution
commenced. After five hours, the mixture was concentrated and the
residue dissolved in dichloromethane (100 mL), cooled to 0.degree.
C. A solution of methyl amine in THF (260 mL of a 2M solution) was
added slowly. After 30 minutes, the mixture was diluted with water
(500 mL) and extracted with chloroform (3.times.). The combined
organic layers were dried (Na.sub.2SO.sub.4), filtered and
concentrated to afford the title compound as a light brown solid.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 6.39 (bs, 1H); 5.80 (d,
1H, J=3.7 Hz); 4.67 (dd, 1H, J=4.0, 3.7 Hz); 4.42 (d, 1H, J=9.3
Hz); 3.6 (dd, 1H, J=9.3, 4.0 Hz); 2.9 (d, 3H, J=5.0 Hz); 1.54 (s,
3H); 1.34 (s, 3H).
Preparation M1
[1155]
(2S,3S,4R,5R)4-Acetoxy-3-azido-5-(6-chloropurin-9-yl)tetrahydrofura-
n-2-carboxylic acid methyl ester
[1156] A solution of 6-chloropurine (4.96 g, 32 mmol) and
hexamethyldisilazane (40 mL) were combined and heated to
100.degree. C. for 3 h under anhydrous conditions. The mixture was
allowed to cool to room temperature and was then concentrated to a
solid on the rotary evaporator using anhydrous toluene (3.times.50
mL) to help remove the solvent. The solid was pumped on high vacuum
for 15 minutes and then anhydrous acetonitrile (50 mL) was added.
1,2-Bis-O-acetyl-3-Azido-3-deox- y-D-ribofuranuronic acid methyl
ester (3.09 g, 10.7 nmol) was dissolved in anhydrous acetonitrile
(20 mL) and added to the reaction.
[1157] Trimethylsilyltrifluoromethanesulfonate (7.5 mL, 41.4 mmol)
was added. The reaction was stirred at 70.degree. C., under
anhydrous conditions, for 15 h. The reaction mixture was quenched
with saturated aqueous sodium bicarbonate (200 mL). Water (160 mL)
was added, and the reaction mixture was extracted with EtOAc
(4.times.100 mL), dried over sodium sulfate, and concentrated to a
solid on the rotary evaporator. This solid was purified via flash
chromatography (7:3 hexane:EtOAc) to afford 2.88 g of the title
compound as a white foam.
[1158] Mp 94.0-96.0.degree. C.
[1159] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.73 (s, 1H); 8.60
(s, 1H); 6.36 (d, 1H, J=5.4 Hz); 5.83 (t, 1H, J=5.4 Hz); 4.83-4.77
(mult, 1H); 4.65 (d, 1H, J=4.2 Hz); 3.84 (s, 3H); 2.14 (s, 3H).
Preparation N1
[1160] 1,2-Bis-O-acetyl-3-azido-3-deoxy-D-ribofuranuronic acid
methyl ester
3-Azido-3-deoxy-1,2-O-isopropylidene-.alpha.-D-ribofuranuronic acid
methyl ester (4.85 g, 20 mmol), concentrated H.sub.2SO.sub.4 (5.5
mL), glacial acetic acid (65 mL), and acetic anhydride (18 mL, 20
mmol) were combined and stirred at room temperature, under
anhydrous conditions, for 15 h. The reaction mixture was then taken
up in water (500 mL), neutralized with solid sodium hydroxide to pH
7 and extracted with EtOAc (3.times.250 mL). The combined organic
layers were washed with saturated aqueous NaCl (500 mL), dried over
sodium sulfate and concentrated on a rotary evaporator to a solid,
which was purified via flash chromatography (2:1 hexane:EtOAc) to
afford 3.09 g of the title compound as a clear, colorless oil.
[1161] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 6.18 (s, 1H); 5.33
(d, 1H, J=5.0 Hz); 4.53 (d, 1H, J=7.5 Hz); 4.44-4.38 (mult, 1H);
3.83 (s, 3H); 2.17 (s, 3H); 2.08 (s, 3H).
Preparation O1
[1162]
3-Azido-3-deoxy-1,2-O-isopropylidene-.alpha.-D-ribofuranuronic acid
methyl ester
[1163] To a solution of
3-azido-3-deoxy-1,2-isopropylidene-.alpha.-D-ribof- uranuronic acid
(4.23 g, 18 mmol) in DMF (50 mL) was added potassium carbonate (3
g, 22 mmol) and iodomethane (2.3 mL, 37 mmol). The reaction mixture
was stirred at room temperature under anhydrous conditions for 15
h. The reaction mixture was then taken up in EtOAc (500 mL) and
washed with water (500 mL), saturated aqueous NaHCO (2.times.500
mL), and saturated aqueous NaCl (500 mL). The combined organic
layers were dried over sodium sulfate and concentrated on the
rotary evaporator to an oil. The product was purified by flash
chromatography (7:3 hexane:EtOAc) to afford 4.45 g of the title
compound as a clear, colorless oil.
[1164] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.89 (d, 1H, J=3.3
Hz); 4.74-4.68 (mult, 1H); 4.53 (d, 1H, J=,9.6 Hz); 3.82 (s, 3H);
3.69-3.63 (mult, 1H); 1.55 (s, 3H); 1.34 (s, 3H).
Preparation P1
[1165] 3-Azido-3-deoxy-1,2-isopropylidene-.alpha.-D-ribofuranuronic
acid
[1166] A solution of
3-azido-1,2,5,6-bis-O-isopropylidene-3-deoxy-D-allofu- ranose (451
g, 1.58 mol) in diethyl ether (4.5 L) was treated with periodic
acid (540 g, 2.37 mol) at ambient temperature which was maintained
with a water bath. After 24 hours, the precipitated salts were
filtered and washed with ether. The filtrate was concentrated and
the crude aldehyde was added to a mixture of chloroform (2.5 L),
acetonitrile (2.5 L) and water (3.4 L). To this vigorously stirred
mixture was added sodium periodate (1211 g, 5.67 mol) and ruthenium
trichloride hydrate (14.5 g, 0.69 mol) at room temperature
maintained with a water bath. After 20 hours, the mixture was
diluted with chloroform (4 L) and water (4 L) and the mixture was
filtered through. Celite.RTM. filter aid. The layers were separated
and the aqueous phase reextracted with chloroform. The combined
organic layers were concentrated in vacuo and the residue
partitioned between saturated aqueous sodium bicarbonate (2 L) and
ethyl acetate (3 L). The layers were separated and the aqueous
layer reextracted with ethyl acetate (2 L). The aqueous layer was
acidified with 2N HCl solution and extracted. with ethyl acetate
(3.times.3 L). The combined organic layers were dried
(Na.sub.2SO.sub.4), filtered and concentrated to give 208 g of the
title compound as an off-white solid pure by tic and NMR. .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 7.5 (bs, 1H); 5.95 (d, 1H,
J=3.7); 4.8 (dd, 1H, J=4.0, 3.7 Hz); 4.6 (d, 1H, J=9.5 Hz); 3.72
(dd, 1H, J=9.5, 4.0 Hz); 1.58 (s, 3H); 1.38 (s, 3H).
Preparation Q1
[1167]
3-Azido-1,2,5,6-bis-O-isopropylidene-3-deoxy-D-allofuranose.
[1168] Triflic anhydride (1500 g, 5.3 mol) was added dropwise to a
solution of pyridine (493 g, 6.2 mol) in dichloromethane (7.5 L) at
-15.degree. C. After 30 minutes, a solution of
1,2,5,6-bis-O-isopropylide- ne-D-glucofuranose (735 g, 2.84 mol)
was added as a solution in dichloromethane (2.5 L). After 1 hour,
the reaction was quenched by the dropwise addition of water (4 L)
allowing the reaction temperature to warm to 0.degree. C. The
layers were separated and the organic layer dried with sodium
sulfate, filtered and concentrated to give a red oil. The triflate
was immediately dissolved in DMF (8 L), treated with NaN.sub.3 (554
g, 8.5 mol) and warmed to 35.degree. C. After 18 hours, the mixture
was poured into water (12 L) and extracted with ethyl acetate
(3.times.4 L). The combined organic layers were washed with water
(2.times.3 L) and brine (1.times.3 L), dried (Na.sub.2SO.sub.4),
filtered and concentrated. The residue was preadsorbed onto silica
gel and purified by flash chromatography (6:1 hex/EtOAc then 4:1
hex/EtOAc) to afford 228 g of the title compound as a colorless
oil. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.77 (d, 1H, J=3.7
Hz); 4.7 (dd, 1H, J=4.4, 3.7 Hz); 4.15 (m, 2H); 4.0 (m, 2H); 3.5
(dd, 1H, J=9.1, 4.4 Hz); 1.56 (s, 3H); 1.47 (s, 3H); 1.37 (s, 3H);
1.34 (s, 3H).
Preparation R1
[1169]
5-Chloro-2-(4,5-dihydro-1H-imidazol-2-ylmethoxy)-benzylamine
[1170] A solution of 30% HCl in ether (0.5 mL) was added to a
solution of
[5-chloro-2-(4,5-dihydro-1H-imidazol-2-ylmethoxy)-benzyl]-carbamic
acid tert-butyl ester (146 mg, 0.43 mmol) in methanol (5 mL). After
stirring overnight, the mixture was concentrated and triturated
from ether, filtered, washed with ether and dried to afford 76 mg
of the title compound as a colorless solid. .sup.1H NMR (400 MHz,
CD.sub.3OD) .delta. 7.4 (m, 2H); 7.04 (m, 1H); 4.91 (s, 2H); 4.12
(s, 2H); 4.00 (s, 2H); 3.79 (s, 2H).
Preparation S1
[1171] (5-Chloro-2-cyanomethoxy-benzyl)-carbamic acid tert-butyl
ester
[1172] Di-tert-butyl dicarbonate (0.45 g, 2.03 mmol) was added to a
solution of (2-aminomethyl-4-chloro-phenoxy)-acetonitrile (0.2 g,
1.01 mmol) and triethylamine (0.57 mL, 4 mmol) in dry THF (10 mL)
at ambient temperature. After 4 hours the mixture was diluted with
ethyl acetate (30 mL) washed with 1N HCl solution (1.times.), sat.
sodium bicarbonate solution (1.times.) and brine (1.times.), dried
(Na.sub.2SO.sub.4), filtered and concentrated. The residue was
purified by flash chromatography (5-20% ethyl acetate/hexanes) to
afford 218 mg (74%) of the title compound as a colorless solid.
[1173] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.3 (m, 2H); 6.92
(d, 1H, J=8.7 Hz); 4.91 (bs, 1H); 4.78 (s, 2H); 4.28 (s, 2H); 1.41
(s, 9H).
Preparation T1
[1174] 2-Aminomethyl4-chloro-phenoxy acetic acid tert-butyl
ester.
[1175] Raney nickel (.about.0.5 g) was added to a solution of
2-cyano-4-chlorophenoxy acetic acid-t-butyl ester (560 mg, 0.21
mmol) in methanol (25 mL) containing 1 mL of sat. ammonium
hydroxide. The mixture was placed under 40 psi of hydrogen and
shaken for two hours. The mixture was filtered and concentrated.
The residue was partitioned between 1N HCl and ether. The layers
were separated and the aqueous layer was re-extracted with ether.
The aqueous layer was basified to .about.pH 10 with K.sub.2
CO.sub.3 solution and extracted with ether (3.times.). The combined
organic layers were dried (MgSO.sub.4), filtered and concentrated
to afford 360 mg of the title compound as an oil. MS 272
(M+H).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.22 (s,
1H); 7.12 (d, 1H, J=8.7 Hz); 6.61 (d, 1H, J=8.7 Hz); 4.5 (s, 2H);
3.85 (s, 2H); 2.62 (bs, 2H); 1.41 (s, 9H).
Preparation U1
[1176] 2-cyano-4-chlorophenoxy acetic acid-tert-butyl ester
[1177] Sodium hydride (96 mg, 4.1 mmol) was added to a solution of
2-cyano-4-chloro phenol (0.5 g, 3.26 mmol) in anhydrous DMF (6 mL)
at -10.degree. C. After 15 minutes the mixture became homogeneous
and t-butyl bromo acetate was added dropwise. After 2 hours, the
mixture was quenched with water and extracted with ethyl acetate
(2.times.). The combined organic layers were washed with water
(2.times.) brine (1.times.), dried (Na.sub.2SO.sub.4), filtered and
concentrated. The residue was purified by flash chromatography (10%
ethyl acetate/hexanes) to afford 850 mg of the title compound as a
colorless syrup (97%). .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.53 (d, 1H, J=2.7 Hz); 7.46 (dd, 1H, J=9.1, 2.7 Hz); 6.78 (d, 1H,
J=9.1 Hz); 4.63 (s, 3H); 1.46 (s, 9H).
Preparation V1
[1178] (-)-5-Chloro-2-(tetrahydrofuran-3-ylmethoxy)benzylamine
[1179] After converting
5-chloro-2-(tetrahydrofuran-3-ylmethoxy)benzylamin- e hydrochloride
into its free base, the free base was subjected to chiral HPLC, to
yield the title compound.
[1180] [.alpha.].sub.22=-23.57.degree. (c=0.140, MeOH)
[1181] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.35 (bs, 3H);
7.49 (d, 1H, J=2.7 Hz); 7.37 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.07 (d,
1H, J=8.7 Hz); 4.00-3.90 (mult, 4H); 3.80-3.70 (mult, 2H);
3.65-3.60 (mult, 1H); 3.60-3.55 (mult, 1H); 2.75-2.71 (mult, 1H);
2.05-1.95 (mult, 1H); 1.77-1.63 (mult, 1H).
[1182] The following compound, Preparation V2, was prepared using
analogous procedures to Preparation V1.
Preparation V2
[1183] (+)-5-Chloro-2-(tetrahydrofuran-3-ylmethoxy)benzylamine
[1184] After converting
5-chloro-2-(tetrahydrofuran-3-ylmethoxy)benzylamin- e hydrochloride
into its free base, the free base was subjected to chiral HPLC, to
yield the title compound. ps [.alpha.].sub.22=+16.32.degree. (c
=0.190, MeOH)
[1185] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.35 (bs, 3H);
7.49 (d, 1H, J=2.7 Hz); 7.37 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.07 (d,
1H, J=8.7 Hz); 4.00-3.90 (mult, 4H); 3.80-3.70 (mult, 2H);
3.65-3.60 (mult, 1H); 3.60-3.55 (mult, 1H); 2.75-2.71 (mult, 1H);
2.05-1.95 (mult, 1H); 1.77-1.63 (mult, 1H).
Preparation W1
[1186] 5-Chloro-2-(2-morpholin-4-yl-ethoxy)benzylamine
hydrochloride
[1187] A solution of
5-chloro-2-(2-morpholin-4-yl-ethoxy)benzonitrile (412 mg, 1.54
mmol) in anhydrous THF (15 mL) was added to a solution of lithium
aluminum hydride (3.53 mL, 1.0 M in THF) at room temperature. The
reaction was stirred for 15 h at room temperature under anhydrous
conditions, whereupon it was cooled to 0.degree. C. and 1 N aqueous
NaOH (0.54 mL) was added. The reaction was stirred for 1/2 h at
0.degree. C. and then the solids were filtered off and rinsed with
THF. The filtrate was concentrated by rotary evaporation and the
resulting solid was dissolved in 30 mL of absolute ethanol. Aqueous
hydrochloride solution (1.0 N, 1.7 mL) was added and the reaction
was stirred for 15 min. The solvent was removed by rotary
evaporation and the resulting solid was triturated with Et.sub.2O
to to yield the title compound.
[1188] C.sub.13H.sub.19ClN.sub.2O.sub.2. MW 270.76. MS 271.2
(M+H).sup.+.
[1189] .sup.1H NMR (400 MHz, CD.sub.3OD) .delta. 7.45-7.40 (mult,
2H); 7.13 (d, 1H, J=5.9); 4.34 (bs, 2H); 4.15 (s, 2H); 3.79 (bs,
4H); 3.35-3.25 (mult, 4H); 3.08 (bs, 2H); 2.83 (bs, 2H)
[1190] The following compounds, Preparations W2-W26, were prepared
from the appropriate benzonitrile using analogous procedures to
Preparation W1.
Preparation W2
[1191] 2-Benzyloxy-5-chloro-benzylamine hydrochloride
[1192] Mp 150.5-152.5.degree. C.
[1193] C.sub.14H.sub.14ClNO. MW 247.73. MS 248.1 (M+H).sup.+.
[1194] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.34 (bs, 3H);
7.49 (d, 1H, J=2.7 Hz); 7.47-7.38 (mult, 2H); 7.43 to 7.33 (mult,
3H) 7.32-7.25 (mult, 1H); 7.10 (d, 1H, J=8.7 Hz); 5.14 (s, 2H);
3.96 (s, 2H).
Preparation W3
[1195] 5-Chloro-2-cyclobutylmethoxybenzylamine hydrochloride
[1196] Mp 168.0-170.0.degree. C.
[1197] C.sub.12H.sub.16ClNO. MW 225.72. MS 225.8 (M+H).sup.+.
[1198] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.36 (bs, 3H);
7.48 (d, 1H, J=2.5 Hz); 7.36 (dd, 1H, J=8.7 Hz, J=2.5 Hz); 7.06 (d,
1H, J=8.7 Hz); 3.97 (d, 2H, J=6.4 Hz); 3.92 (bs, 2H); 2.76-2.71
(mult, 1H); 2.09-1.97 (mult, 2H); 1.97-1.77 (mult, 4H).
Preparation W4
[1199] 5-Chloro-2-(3-methoxy-benzyloxy)benzylamine
hydrochloride.
[1200] Mp 159.0-160.0.degree. C.
[1201] C.sub.15H.sub.16ClNO.sub.2. MW 277.75. MS 277.9
(M+H).sup.+.
[1202] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.37 (bs, 3H);
7.51 (d, 1H, J=2.7 Hz); 7.38 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.28 (t,
1H, J=8.0 Hz); 7.10 (d, 1H, J=8.9 Hz); 7.06 7.00 (mult, 2H); 6.87
(d, 1H, J=8.3 Hz); 5.13 (s, 2H); 3.99 (s, 2H); 3.74 (s, 3H).
Preparation W5
[1203] 5-Chloro-2-(2,5-dimethoxy-benzyloxy)benzylamine
hydrochloride
[1204] Mp 212.0-213.0.degree. C.
[1205] C.sub.16H.sub.18ClNO.sub.3. MW 307.78. MS 307.8
(M+H).sup.+.
[1206] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.30 (bs, 3H);
7.50 (d, 1H, J=2.7 Hz); 7.38 (dd, 1H, J=8.9 Hz, J=2.7 Hz); 7.11 (d,
1H, J=8.9 Hz); 7.02 (d, 1H, J=3.1 Hz); 6.96 (d, 1H, J=8.9 Hz); 6.86
(dd, 1H, J=8.9 Hz, J=3.1 Hz); 5.09 (s, 2H); 3.98 (s, 2H); 3.75 (s,
3H); 3.68 (s, 3H).
Preparation W6
[1207] 5-Chloro-2-(3-chloro-benzyloxy)benzylamine hydrochloride
[1208] Mp 164.0-166.0.degree. C.
[1209] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.31 (bs, 3H);
7.58 (s 1H); 7.52 (d, 1H, J=2.7 Hz); 7.50-7.37 (mult, 4H); 7.11 (d,
1H, J=8.9 Hz); 5.19 (s, 2H); 4.02 (bs, 2H).
Preparation W7
[1210] 5-Chloro-2-(2-chloro-benzyloxy)benzylamine hydrochloride
[1211] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.33 (bs, 3H);
7.85-7.80 (mult, 1H); 7.55-7.45 (mult, 2H); 7.45-7.35 (mult, 3H);
7.17 (d, 1H, J=8.7 Hz); 5.22 (s, 2H); 4.00 (d, 2H, J=5.2 Hz).
Preparation W8
[1212] 5-Chloro-2-(tetrahydrofuran-3-ylmethoxy)benzylamine
hydrochloride
[1213] Mp 105.5-108.0.degree. C.
[1214] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.35 (bs, 3H);
7.49 (d, 1H, J=2.7 Hz); 7.37 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.07 (d,
1H, J=8.7 Hz); 4.00-3.90 (mult, 4H); 3.80-3.70 (mult, 2H);
3.65-3.60 (mult, 1H); 3.60-3.55 (mult, 1H); 2.75-2.71 (mult, 1H);
2.05-1.95 (mult, 1H); 1.77-1.63 (mult, 1H).
Preparation W9
[1215] 5-Chloro-2-(4-methyl-benzyloxy)benzylamine hydrochloride
[1216] Mp 155.0-157.0.degree. C.
[1217] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.33 (bs, 3H);
7.50 (d, 1H, J=2.7 Hz); 7.40-7.35 (mult, 3H); 7.17 (d, 2H, J=7.7
Hz); 7.12 (d, 1H, J=8.9 Hz); 5.11 (s, 2H); 3.96 (bs, 2H); 2.28 (s,
3H).
Preparation W10
[1218] 5-Chloro-2-(2-methyl-benzyloxy)benzylamine hydrochloride
[1219] Mp 176.0-178.0.degree. C.
[1220] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.38 (bs, 3H);
7.53 (d, 1H, J=2.7 Hz); 7.45-7.38 (mult, 2H); 7.25-7.17 (mult, 4H);
5.15 (s, 2H); 3.97 (d, 2H, J=5.4 Hz); 2.31 (s, 3H).
Preparation W11
[1221] 5-Chloro-2-(3-methyl-benzyloxy)benzylamine hydrochloride
[1222] Mp 137.0-140.0.degree. C.
[1223] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.26 (bs, 3H);
8.49 (d, 1H, J=2.5 Hz); 7.39 (dd, 1H, J=8.7 Hz, J=2.7 Hz);
7.29-7.22 (mult, 3H); 7.12 (d, 2H, J=8.9 Hz); 5.12 (s, 2H);
4.02-3.97 (mult, 2H); 2.30 (s, 3H).
Preparation W12
[1224] 5-Chloro-2-(2-methoxy-benzyloxy)benzylamine
hydrochloride
[1225] Mp. 199.5-200.5.degree. C.
[1226] C.sub.15H.sub.16ClNO.sub.2. MW 277.75. MS 278.1
(M+H).sup.+.
[1227] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.37 (bs, 3H);
7.51 (d, 1H, J=2.3 Hz); 7.41 (d, 1H, J=7.5 Hz); 7.37 (dd, 1H, J=8.7
Hz, J=2.5 Hz); 7.33 (dd, 1H, J=8.3 Hz, J=7.5 Hz); 7.12 (d, 1H,
J=8.9 Hz); 7.04 (d, 1H, J=8.3 Hz); 6.94 (t, 1H, J=7.5Hz); 5.12 (s,
2H); 3.97 (bs, 2H); 3.81 (s, 3H).
Preparation W13
[1228] 5-Chloro-2-(furan-3-ylmethoxy)benzylamine hydrochloride
[1229] Mp 137.0-140.0.degree. C.
[1230] C.sub.12H.sub.12ClNO.sub.2. MW 237.7. MS 238.1
(M+H).sup.+.
[1231] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.33 (bs, 3H);
7.80 (s, 1H); 7.66 (s,1H); 7.49 (d, 1H, J=2.7 Hz); 7.39 (dd, 1H,
J=8.9 Hz, J=2.7 Hz); 7.17 (d, 1H, J=8.7 Hz); 6.62 (s, 1H); 5.03 (s,
2H); 3.92 (quart, 2H, J=5.7 Hz).
Preparation W14
[1232] 5-Chloro-2-(4-methoxy-benzyloxy)benzylamine
[1233] Mp 142.0-147.0.degree. C.
[1234] C.sub.15H.sub.16ClNO.sub.2. MW 277.75. MS 278.1
(M+H).sup.+.
[1235] .sup.1H NMR (400 MHz, DMSO) .delta. 8.38 (bs, 2H); 8.25 (bs,
1H); 7.50 (d, 1H, J=2.1 Hz); 7.43-7.36 (mult, 3H); 7.14 (d, 1H,
J=8.7 Hz); 6.95-6.90 (mult, 2H); 5.07 (s, 2H); 3.96-3.90 (mult,
2H); 3.73 (s, 3H).
Preparation W15
[1236] 5-Chloro-2-cyclopentylmethoxybenzylamine
[1237] Mp 155.0-157.0.degree. C.
[1238] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.34 (bs, 3H);
7.47 (d, 1H, J=2.5 Hz); 7.36 (dd, 1H, J=8.9 Hz, J=2.5 Hz); 7.06 (d,
1H, J=8.9 Hz); 3.94 (bs, 2H); 3.88 (d, 2H, J=6.8 Hz); 2.32 (sept,
1H, J=7.5 Hz); 1.80-1.70 (mult, 2H); 1.62-1.45 (mult, 4H);
1.38-1.28 (mult, 2H).
Preparation W16
[1239] 5-Chloro-2-[3-(2-morpholin-4-yl-ethoxy)benzyloxy]benzylamine
hydrochloride
[1240] C.sub.20H.sub.25ClN.sub.2O.sub.3. MW 376.89. MS 377.1
(M+H).sup.+.
[1241] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.39 (bs, 3H);
7.51 (d, 1H, J=2.7 Hz); 7.39 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.31 (t,
1H, J=7.9 Hz); 7.12 (d, 2H, J=8.9 Hz); 7.07 (d, 1H, J=7.5 Hz); 6.93
(d, 1H, J=8.5 Hz); 5.14 (s, 2H); 4.47-4.27 (mult, 2H); 4.02-3.98
(mult, 2H); 3.98-3.70 (mult, 4H); 3.59-3.39 (mult, 4H); 3.35-3.25
(mult, 2H).
Preparation W17
[1242] 5-Chloro-2-(tetrahydrofuran-3-ylmethoxy)benzylamine
hydrochloride
[1243] Mp 105.5-108.0.degree. C.
[1244] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.35 (bs, 3H);
7.49 (d, 1H, J 2.7 Hz); 7.37 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.07 (d,
1H, J=8.7 Hz); 4.00-3.90 (mult, 4H); 3.80-3.70 (mult, 2H);
3.65-3.60 (mult, 1H); 3.60-3.55 (mult, 1H); 2.75-2.71 (mult, 1H);
2.05-1.95 (mult, 1H); 1.77-1.63 (mult, 1H).
Preparation W18
[1245] 5-Chloro-2-(tetrahydrofuran-3-ylmethoxy)benzylamine
hydrochloride
[1246] Mp 105.5-108.0.degree. C.
[1247] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.35 (bs, 3H);
7.49 (d, 1H, J=2.7 Hz); 7.37 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.07 (d,
1H, J=8.7 Hz); 4.00-3.90 (mult, 4H); 3.80-3.70 (mult, 2H);
3.65-3.60 (mult, 1H); 3.60-3.55 (mult, 1H); 2.75-2.71 (mult, 1H);
2.05-1.95 (mult, 1H); 1.77-1.63 (mult, 1H).
Preparation W19
[1248] 5-Chloro-2-(furan-2-ylmethoxy)benzylamine hydrochloride
[1249] Mp 145.0-150.0.degree. C.
[1250] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.35 (bs, 3H);
7.69 (s, 1H); 7.50 (d, 1H, J=2.5 Hz); 7.39 (dd, 1H, J=8.9 Hz, J=1.9
Hz); 7.25 (d, 1H, J=8.5 Hz); 6.60 (d, 1H, J=3.1 Hz);. 6.46 (bs,
1H); 5.14 (s, 2H); 3.89 (d, 2H, J=4.4 Hz).
Preparation W20
[1251]
5-Chloro-2-(2,2,7,7-tetramethyltetrahydro-bis[1,3]dioxolo[4,5-b;4',-
5'-d]pyran-5-ylmethoxy)benzylamine hydrochloride
[1252] .sup.1H NMMR (400 MHz, DMSO-d.sub.6) .delta. 8.23 (bs, 3H);
7.48-7.42 (mult, 1H); 7.34 (dd, 1H, J=8.9 Hz, J=2.7 Hz); 7.13 (d,
1H, J=8.9 Hz); 5.43 (d, 1H, J=5.0 Hz); 4.59 (dd, 1H, J=8.1 Hz,
J=2.3 Hz); 4.36-4.40 (mult, 2H); 4.01 (s, 2H); 4.05-3.80 (mult,
3H); 1.30 (d, 6H, J=6.6 Hz); 1.23 (d, 6H, J=11.0 Hz).
Preparation W21
[1253]
5-Chloro-2-(5-dimethylaminomethylfuran-2-ylmethoxy)benzylamine
[1254] C.sub.15H.sub.19ClN.sub.2O.sub.2. MW 294.78. MS 295.2
(M+H).sup.+.
[1255] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 8.49 (bs, 3H);
7.52 (d, 1H, J=2.4 Hz); 7.43 (dd, 1H, J=9.0 Hz, J=2.4 Hz); 7.29 (d,
1H, J=9.0 Hz); 6.77-6.68 (mult, 2H); 5.18 (s, 2H); 4.39-4.34 (mult,
2H); 3.93 (d, 2H, J =5.6 Hz); 2.76-2.61 (mult, 6H).
Preparation W22
[1256] 5-Chloro-2-(naphthalen-1-ylmethoxy)benzylamine
hydrochloride
[1257] Mp 196.0-198.0.degree. C.
[1258] C.sub.18H.sub.16ClNO. MW 297.79. MS 297.9 (M+H).sup.+.
[1259] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 5 8.35 (bs, 3H);
8.14 (dd, 1H, J=8.9 Hz, J=2.7 Hz); 8.03-7.92 (mult, 2H); 7.72 (d,
1H, J=6.9 Hz); 7.63-7.41 (mult, 5H); 7.40 (d, 1H, J=8.9 Hz); 5.66
(s, 2H); 3.95 (d, 2H, J=5.6 Hz).
Preparation W23
[1260] 5-Chloro-2-(4-chloro-benzyloxy)benzylamine hydrochloride
[1261] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.35 (bs, 3H);
7.54-7.47 (mult, 3H); 7.46-7.35 (mult, 3H); 7.10 (d, 1H, J=8.9 Hz);
5.16 (s, 2H); 3.99 (bs, 2H).
Preparation W24
[1262] 5-Chloro-2-(2,5-dimethylfuran-3-ylmethoxy)benzylamine
[1263] Using a modified Preparation W1, wherein instead of making
the hydrochloride salt, the title compound was isolated as the free
base 5-chloro-2-(2,5-dimethylfuran-3-ylmethoxy)benzonitrile was
converted to the title compound. Thus, after the NaOH quench and
filter, the solids were washed with THF. The filtrate was
concentrated on the rotary evaporator to yield the title compound
as a pale yellow oil.
[1264] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.31 (d, 1H,
J=2.7 Hz); 7.15 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 6.98 (d, 1H, J=8.7
Hz); 6.00 (s, 1H); 4.78 (s, 2H); 3.56 (s, 2H); 2.18 (s, 3H); 2.14
(s, 3H); 1.65 (bs, 2H).
Preparation W25
[1265] 5-Chloro-2-methoxybenzylamine
[1266] Using a modified Preparation W1, wherein instead of making
the hydrochloride salt, the title compound was isolated as the free
base, 5-chloro-2-methoxybenzonitrile and was converted to the title
compound. Thus, after the NaOH quench and filter, the solids were
washed with THF. The filtrate was concentrated on the rotary
evaporator. The resulting viscous oil was then taken up in
Et.sub.2O (100 mL) and water (100 mL). After separating the layers,
the aqueous layer was extracted twice more with Et.sub.2O. The
combined organic layers were washed with aqueous saturated NaCl,
dried over Na.sub.2SO.sub.4, filtered, and the solvent removed by
rotary evaporation to yield the title compound as a pale yellow
oil.
[1267] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.43-7.38 (mult,
1H); 7.25-7.15 (mult, 1H); 6.92 (td, 1H, J=11 Hz, J=8.7 Hz ); 3.75
(d, 3H, J=7.2 Hz); 3.65-3.55 (mult, 2H); 1.72 (bs, 2H).
Preparation W26
[1268] [3-(2-Morpholin4-yl-ethoxy)phenyl]methanol
[1269] Using a modified Preparation W1, wherein instead of adding
the aqueous hydrochloride solution, the title compound was
extracted as the alcohol, 3-(2-morpholin4-yl-ethoxy)benzaldehyde
and was converted to the title compound. Thus, after the NaOH
quench and filter, the solids were washed with THF. The filtrate
was concentrated on the rotary evaporator. The resulting viscous
oil was then taken up in Et.sub.2O (100 mL) and water (100 mL).
After separating the layers, the aqueous layer was extracted twice
more with Et.sub.2O. The combined organic layers were washed with
aqueous saturated NaCl, dried over MgSO.sub.4, filtered, and the
solvent removed by rotary evaporation to yield the title compound
as a pale yellow oil.
[1270] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.18 (t, 1H, J
7.9 Hz); 6.87-6.81 (mult, 2H); 6.76 (d, 1H, J=8.1 Hz); 5.17-5.09
(mult, 1H); 4.43 (d, 2H, J=5.6 Hz); 4.04 (t, 2H, J=5.6 Hz);
3.59-3.51 (mult, 4H); 2.70-2.62 (mult, 2H); 2.50-2.38 (mult,
4H).
Preparation X1
[1271] 5-Chloro-2-(3-methylisoxazol-5-ylmethoxy)benzylamine
[1272] To a flask containing anhydrous 2-propanol (40 mL),
hydrazine hydrate (4 mL, and anhydrous THF (60 mL), at room
temperature, was added 2-{5-chloro-2-(3-methyl isoxazol-5ylmethoxy)
benzyl]isoindole-1,3-dione (5 g, 13 mmol). The reaction was stirred
at 50.degree. C. for 3 h. The solids were filtered and washed with
THF. The filtrate was concentrated, dissolved in ether (100 ml) and
washed with 1N NaOH (1X) and brine, dried (MgSO.sub.4), filtered
and concentrated to afford the title product as a solid.
[1273] Mp 32.0-35.0.degree. C.; C.sub.12H.sub.13ClN.sub.2O.sub.2.
MW 252.70. MS 253.1 (M+H).sup.+.
[1274] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.37 (d, 1H,
J=2.6 Hz); 7.18 (dd, 1H, J=8.8 Hz, J=2.6 Hz); 7.04 (d, 1H, J=8.8
Hz); 6.42 (s, 1H); 5.21 (s, 2H); 3.60 (s, 2H); 2.18 (s, 3H); 1.71
(bs, 2H).
[1275] The following compounds, Preparations X1-X6, were prepared
from the appropriate phthalimide using analogous procedures to
Preparation X1.
Preparation X2
[1276] 5-Chloro-2-(pyridin-3-ylmethoxy)benzylamine
[1277] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.63 (d, 1H,
J=1.5 Hz); 8.50 (dd, 1H, J=4.8 Hz, J=1.5 Hz); 7.83 (dd, 1H, J=6.0
Hz, J=3.1 Hz); 7.42-7.35 (mult, 2H); 7.20 (dd, 1H, J=8.7 Hz, J=2.9
Hz); 7.02 (d, 1H, J=8.7 Hz); 5.13 (s, 2H); 3.67 (s, 2H).
Preparation X3
[1278] 5-Chloro-2-(thiazol-2-ylmethoxy)benzylamine
[1279] Mp 74.0-76.0.degree. C.
[1280] C.sub.11H.sub.11ClN.sub.2OS. MW 254.74. MS 255.1
(M+H).sup.+.
[1281] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.80 (d, 1H,
J=3.3 Hz); 7.74 (d, 1H, J=3.3 Hz); 7.39 (d, 1H, J=2.7 Hz); 7.19
(dd, 1H, J=8.3 Hz, J=2.7 Hz); 7.05 (d, 1H, J=8.7 Hz); 5.41 (s, 2H);
3.68 (s, 2H); 1.78 (bs, 2H).
Preparation X4
[1282] 2-(Benzothiazol-2-ylmethoxy)-5-chlorobenzylamine
[1283] Mp 72.0-79.0
[1284] C.sub.15H.sub.13ClN.sub.2OS. MW 304.80. MS 305.1
(M+H).sup.+.
[1285] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.09 (d, 1H,
J=7.9 Hz); 7.97 (d, 1H, J=7.9 Hz); 7.49 (dd, 1H, J=8.1 Hz, J=7.3
Hz); 7.44-7.39 (mult, 2H); 7.20 (dd, 1H, J=8.6 Hz, J=2.6 Hz); 7.07
(d, 1H, J=8.6 Hz); 5.57 (s, 2H); 3.75 (s, 2H); 1.83 (bs, 2H).
Preparation X5
[1286] 2-(Benzofuran-2-ylmethoxy)-5-chlorobenzylamine
[1287] Mp 75.5-77.0.degree. C.
[1288] C.sub.16H.sub.14ClNO.sub.2. MW 287.75. MS 288.0
(M+H).sup.+.
[1289] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.60 (d, 1H,
J=7.9 Hz); 7.54 (d, 1H, J=7.9 Hz); 7.36 (d, 1H, J=2.1 Hz); 7.28 (t,
1H, J=7.3 Hz); 7.25-7.16 (mult, 2H); 7.13 (d, 1H, J=8.7 Hz); 6.99
(s, 1H); 5.25 (s, 2H); 3.61 (s, 2H); 1.78 (bs, 2H).
Preparation X6
[1290] 5-Chloro-2-(thiophen-2-ylmethoxy)benzylamine
[1291] C.sub.12H.sub.12ClNOS. MW 253.75. MS 254.0 (M+H).sup.+.
[1292] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.53 (dd, 1H,
J=5.0 Hz, J=1.2 Hz); 7.40-7.36 (mult, 1H); 7.22-7.16 (mult; 2H);
7.07 (dd, 1H, J=8.7 Hz, J=2.3 Hz); 7.04-7.36 (mult, 1H); 5.29 (s,
2H); 3.62 (s, 2H); 1.79 (bs, 2H).
Preparation Y1
[1293] 5-Chloro-2-(isothiazol-5-ylmethoxy)benzylamine
[1294] To a mixture of
2-[5-chloro-2-(isothiazol-5ylmethoxy)benzyl]isoindo- le-1,3-dione
(464 mg, 1.20 mmol), 2-propanol (20 mL), water (2.5 mL), and THF
(25 mL) was added NaBH.sub.4 (295 mg, 7.80 mmol). The reaction was
stirred at room temperature for 16 h. The organic solvent was then
removed by rotary evaporation and the resulting oil was taken up in
CH.sub.2Cl.sub.2 (50 mL). The organic layer was washed with water
(50 mL), saturated aqueous NaCl (50 mL), dried over NaSO.sub.4,
filtered, and concentrated on a rotary evaporator. This material
was taken up in 2-propanol (20 mL), glacial acetic acid (1.2 mL),
and water (1.0 mL) and the reaction was heated to 80.degree. C. in
a sealed tube for 24 h. The organic solvent was then removed by
rotary evaporation and the resulting oil was taken up in
CH.sub.2Cl.sub.2 (50 mL). The organic layer was washed with water
(50 mL), saturated aqueous NaCi (50 mL), dried over NaSO.sub.4,
filtered, and concentrated on a rotary evaporator to afford the
title compound.
[1295] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.50 (s, 1H);
7.42-7.38 (mult, 2H); 7.19 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.02 (d,
1H, J=8.7 Hz); 5.50 (s, 2H); 3.65 (s, 2H); 1.73 (bs, 2H).
[1296] The following compounds, Preparations Y2-Y3, were prepared
from the appropriate azide using analogous procedures to
Preparation Y1.
Preparation Y2
[1297] 5-Chloro-2-(quinolin-2-ylmethoxy)benzylamine
[1298] C.sub.17H.sub.15ClN.sub.2O. MW 298.77. MS 299.2
(M+H).sup.+.
[1299] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.40 (d, 1H,
J=8.5 Hz); 8.03-7.96 (mult, 2H); 7.76 (t, 1H, J=6.8 Hz); 7.65 (d,
1H, J=8.3 Hz); 7.60 (t 1H, J=6.8 Hz); 7.42 (s, 1H); 7.18 (d, 1H,
J=8.5 Hz); 7.01 (d, 1H, J=8.9 Hz); 5.37 (s, 2H); 3.78 (s, 2H).
Preparation Y3
[1300] 5-Chloro-2-(4-methyl-[1,2,3]thiadiazol-5-ylmethoxy)
benzylamine
[1301] Mp 64.0-66.0.degree. C.
[1302] C.sub.11H.sub.12ClN.sub.3OS. MW 269.75. MS 270.1
(M+H).sup.+.
[1303] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.42 (d, 1H,
J=2.3 Hz); 7.25 (dd, 1H, J=8.7 Hz, J=2.3 Hz); 7.10 (d, 1H, J=8.7
Hz); 5.53 (s, 2H); 3.65 (s, 2H); 2.66 (s, 3H); 1.84 (bs, 2H).
Preparation Z1
[1304] 2-Benzyloxy-5-cyano-benzylamine
[1305] Triphenylphosphine (1.64 g, 6.24 mmol) was added to a
solution of 2-benzyloxy-5-cyano-benzylazide (1.1 g, 4.16 mmol) in
anhydrous THF (10 mL) at 0.degree. C. After 1 hour, ammonium
hydroxide (0.5 mL) was added and the mixture was warmed to room
temperature and stirred overnight. The mixture was concentrated and
the residue purified by flash chromatography (3%
methanol/dichloromethane) to afford 677 mg (69%) of the title
compound as a colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.6-7.3 (m, 7H); 6.95 (d, 1H, J=9.1 Hz); 5.18 (s, 2H); 3.9
(s, 2H).
[1306] The following compounds, Preparations Z2-Z4, were prepared
from the appropriate azide using analogous procedures to
Preparation Z1.
Preparation Z2
[1307] 2-Benzyloxy-5-ethenyl-benzylamine
Preparation Z3
[1308] 2-Benzyloxy-5-ethynyl-benzylamine
Preparation Z4
[1309] (2-Aminomethyl4-chloro-phenoxy)-acetonitrile
Preparation AA1
[1310] 2-Benzyloxy-5-trifluoromethyl-benzylamine LAH (136 mg, 3.58
mmol) was added to a solution of
2-benzyloxy-5-trifluoromethyl-benzylazide (1.1 g, 3.58 mmol) at
0.degree. C. After 1 hour, the reaction was quenched by the
sequential addition of water (136 .mu.L), 15% NaOH (136 .mu.L), and
water (400 .mu.L), the mixture was diluted with dichloromethane and
dried with MgSO.sub.4, filtered and concentrated. The residue was
purified by flash chromatography (3% methanol/dichloromethane) to
afford 825 mg (82%) of the title compound as a colorless oil. MS
282 (M+H).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.6-7.3
(m, 7H); 6.92 (d, 1H, J=9.1 Hz); 5.18 (s, 2H); 3.9 (s, 2H); 2.65
(bs, 2H).
[1311] The following compounds, Preparations AA2-M5, were prepared
from the appropriate azide using analogous procedures to
Preparation AA1.
Preparation AA2
[1312] 2-Benzyloxy-5-bromo-benzylamine
Preparation AA3
[1313] 2-Benzyloxy-5-fluoro-benzylamine
Preparation AA4
[1314] 2-Benzyloxy-5-iodo-benzylamine
Preparation AA5
[1315] 2-Benzyloxy-5-methyl-benzylamine
Preparation BB1
[1316] 2-Benzyloxy-5-trifluoromethyl-benzylazide
[1317] DBU (1 mL, 6.62 mmol) and diphenyl phosphoryl azide (1.6 mL,
7.5 mmol) were added to a solution of
2-benzyloxy-5-trifluoromethyl-benzyl alcohol (1.24 g, 4.41 mmol) in
anhydrous toluene (10 mL) at room temperature. After 3 hours, water
was added and the mixture extracted with ethyl acetate.(3.times.).
The combined organic layers were washed with 1N HCl (1.times.),
brine (1.times.), dried (Na.sub.2SO.sub.4), filtered and
concentrated in vacuo. The residue was purified by flash
chromatography (3% ethyl acetate/hexanes) to afford 1.14 grams of
the title compound as a colorless oil (84%). MS 280
(M-N.sub.2).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.6-7.3 (m, 7H); 7.01 (d, 1H, J=9.1 Hz); 5.18 (s, 2H); 4.42 (s,
2H).
[1318] The following compounds, Preparations BB2-BB7, were prepared
from the appropriate alcohol using analogous procedures to
Preparation BB1.
Preparation BB2
[1319] 2-Benzyloxy-5-bromo-benzylazide
Preparation BB3
[1320] 2-Benzyloxy-5-fluoro-benzylazide
Preparation BB4
[1321] 2-Benzyloxy-5-methyl-benzylazide
Preparation BB5
[1322] 2-Benzyloxy-5-cyano-benzylazide
Preparation BB6
[1323] 2-Benzyloxy-5-iodo-benzylazide
Preparation BB7
[1324] (2-Azidomethyl-4-chloro-phenoxy)-acetonitrile
Preparation CC1
[1325] 2-Benzyloxy-5-trifluoromethyl-benzylalcohol
[1326] Cesium carbonate (2.24 g, 6.9 mmol) was added to a solution
of 2-hydroxy-5-trifluoromethyl-benzyl alcohol (885 mg, 4.6 mmol)
and benzyl bromide (547 .mu.L, 4.6 mmol) in anhydrous DMF (8 mL) at
room temperature. The reaction was heated to 50.degree. C. for 2
hours, cooled, diluted with ethyl acetate (25 mL) and washed with
water (3.times.25 mL), brine (1.times.25 mL), and dried
(Na.sub.2SO.sub.4), filtered and concentrated to afford 1.3 g of
the title compound as a colorless solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.6-7.3 (m, 7H); 7.01 (d, 1H, J=9.1 Hz); 5.18
(s, 2H); 4.79 (s, 2H); 2.8 (bs, 1H).
Preparation DD1
[1327] 2-Hydroxy-5-trifluoromethyl-benzyl alcohol
[1328] Paraformaldehyde (3.4 g, 0.114 mol) was added in 0.5 9
portions over a six hour period to a refluxing mixture of
4-trifluoromethyl phenol (2.3 g, 0.0142 mol), phenyl boronic acid
(2.1 g, 0.017 mol) and proprionic acid (530 ML, 7 mmol) in benzene
with the azeotropic removal of water (Dean-Stark trap). When the
addition was complete, the reaction was heated for an additional
hour, then cooled with an ice bath, diluted with THF (30 mL) and
treated with 5 mL of 30% hydrogen peroxide solution. After stirring
for one hour, the mixture was partitioned between water (50 mL) and
ethyl acetate (50 mL). The organic layer was washed with
NaHSO.sub.3 solution (1.times.50 mL), brine (1.times.50 mL), dried
(Na.sub.2SO.sub.4), filtered and concentrated. The residue was
purified by flash chromatography (30% ethyl acetate/hexanes) to
afford 950 mg (35%) of the title compound as a colorless solid.
.sup.1H NMR (400 MHz, CDCl.sub.3) 8 7.42 (d, 1H, J=9.1 Hz); 7.3 (s,
1H); 6.95 (d, 1H, J=9.1 Hz); 4.89 (s, 2H);
[1329] The following compound, Preparation DD2, was prepared from
the appropriate phenol using analogous procedures to Preparation
DD1.
Preparation DD2
[1330] 2-Hydroxy-5-cyano-benzyl alcohol
Preparation EE1
[1331] 2-Benzyloxy-5-fluoro benzyl alcohol
[1332] LAH (226 mg, 5.95 mmol) was added to a solution of
2-benzyloxy-5-fluoro-benzoic acid benzyl ester (2 g, 5.95 mmol) at
0.degree. C. The reaction was warmed to room temperature and after
1 hour, the reaction was quenched by the sequential addition of
water (226 .mu.L), 15% NaOH (226 .mu.L), and water (760 .mu.L), the
mixture was diluted with ethyl acetate and dried with MgSO.sub.4,
filtered and concentrated. The residue was purified by flash
chromatography (10% ethyl acetate/hexanes) to afford 1.13 g (82%)
of the title compound as a colorless solid. MS 250 (M+NH4).sup.+.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.4 (m, 5H); 7.05 (dd,
1H, J 9.1, 4.3 Hz); 6.92 (m, 2H); 5.08 (s, 2H); 4.65 (s, 2H); 2.2
(bs, 1H).
[1333] The following compounds, Preparations EE2-EE3, were prepared
from the appropriate starting material using analogous procedures
to Preparation EE1.
Preparation EE2
[1334] 2-Benzyloxy-5-iodo-benzyl alcohol
Preparation EE3
[1335] 2-Benzyloxy-5-methyl-benzyl alcohol
Preparation FF1
[1336] 2-Benzyloxy-5-ethynyl-benzyl azide and
2-benzyloxy-5-ethenyl-benzyl azide
[1337] Lithium aluminum hydride (95 mg, 2.49 mmol) was added to a
solution of 2-benzyloxy-5-ethynyl-benzoic acid benzyl ester (680
mg, 1.99 mmol) in THF (5 mL) at 0.degree. C. After 15 minutes, the
reaction was quenched by the sequential addition of water (100
.mu.L), 15% NaOH (100 .mu.L), and water (300 .mu.L). The mixture
was diluted with ethyl acetate (10 mL) and dried (MgSO.sub.4),
filtered, and concentrated. The residue was purified by flash
chromatography (10% ethyl acetate/hexanes) to afford 600 mg of a
.about.1:1 mixture of 2-benzyloxy-5-ethynyl-benzyl alcohol and
2-benzyloxy-5-ethenyl-benzyl alcohol. This mixture was dissolved in
toluene (8 mL) and treated with DBU (0.7 mL, 4.52 mmol) and
diphenyl phosphoryl azide (1.1 mL, 5.12 mmol) at room temperature.
After 18 hours, water was added and the mixture was extracted with
ethyl acetate (3.times.). The combined organic layers were washed
with 1N HCl (1.times.), brine (1.times.), dried (Na.sub.2SO.sub.4),
filtered and concentrated in vacuo. The residue was purified by
flash chromatography (3.5% THF/hexanes) and the mixed fractions
were rechromatographed with 3% ether/pet ether to afford 206 mg of
2-benzyloxy-5-ethenyl-benzyl azide as the faster eluting fraction.
MS 238 (M-N.sub.2).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.4 (m, 7H); 6.95 (d, 1H, J=9.1 Hz); 6.62 (dd, 1H, J=17.5, 10.5
Hz); 5.61 (d, 1H, J=17.5 Hz); 5.15 (m, 3H); 4.40 (s, 2H).
[1338] Further elution provided 135 mg of
2-benzyloxy-5-ethynyl-benzyl azide. MS 236 (M--N.sub.2).sup.30 .
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.4 (m, 7H); 7.95 (d, 1H,
J=9.1 Hz); 5.1 (s, 2H); 4.39 (s, 2H); 3.0 (s, 1H).
Preparation GG1
[1339] 2-Benzyloxy-5-ethynyl-benzoic acid benzyl ester
[1340] Tetra-butyl ammonium fluoride (2.97 mL of a 1M solution in
THF, 2.97 mmol) was added to a solution of
2-benzyloxy-5-(2-trimethylsilyl-eth- ynyl)-benzoic acid benzyl
ester (1.07 g, 2.58 mmol) in THF (10 mL) at room temperature. After
30 minutes, the mixture was diluted with ether (20 mL) and washed
with water (3.times.30 mL) and brine (1.times.30 mL), dried
(MgSO.sub.4), filtered and concentrated. The residue was purified
by flash chromatography (5% ethyl acetate/hexanes) to afford 687 mg
(78%) product as a light orange oil. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.98 (d, 1H, J=2 Hz); 7.52 (dd, 1H, J=9.1, 1
Hz); 7.38 (m, 10H); 6.95 (d, 1H, J=9.1 Hz); 5.35 (s, 2H); 5.19 (s,
2H); 3.0 (s, 1H).
Preparation HH1
[1341] 2-Benzyloxy-5-(2-trimethylsilyl-ethynyl)-benzoic acid benzyl
ester
[1342] Pd(PPh.sub.3).sub.2Cl.sub.2 (194 mg, 0.277 mmol), copper
iodide (106 mg, 0.554 mmol) and triethylamine (0.8 mL, 5.44 mmol)
were added to a solution of 2-benzyloxy-5-iodo-benzoic acid benzyl
ester (1.23 g, 2.77 mmol) and trimethylsilyl acetylene (0.5 mL,
3.32 mmol) in anhydrous DMF (15 mL) at room temperature. The flask
was covered with aluminum foil and stirred overnight. The mixture
was diluted with ethyl acetate (50 mL) and washed with water
(3.times.), 1N HCl (1.times.) and brine (1.times.), dried
(Na.sub.2SO.sub.4), filtered and concentrated. The residue was
purified by flash chromatography (5% ethyl acetate/hexanes) to
afford 1.07 g (93%) of the title compound as an oil. .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.98 (d, 1H, J=2 Hz); 7.52 (dd, 1H,
J=9.1, 1 Hz); 7.38 (m, 10H); 6.95 (d, 1H, J=9.1 Hz); 5.31 (s, 2H);
5.17 (s, 2H); 0.1 (s, 9H).
Preparation II1
[1343] 2-Benzyloxy-5-fluoro-benzoic acid benzyl ester.
[1344] Benzyl bromide (0.86 mL, 7.2 mmol) and cesium carbonate (2.6
g, 8 mmol) were added to a solution of 2-hydroxy-5-fluoro-benzoic
acid (0.5 g, 3.2 mmol) in anhydrous DMF (8 mL) at room temperature.
The reaction was heated to 80.degree. C. for two hours, cooled and
diluted with water (50 mL). The mixture was extracted with ethyl
acetate (3.times.40 mL) and the combined organic layers were washed
with water (1.times.50 mL) and brine (1.times.50 mL), dried
(Na.sub.2SO.sub.4), filtered and concentrated to afford 1.02 g of
the title compound as a colorless oil (93%). MS: 337 (M+H).sup.+.
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.55 (dd, 1H, J=8.7, 3.1
Hz); 7.35 (m, 10OH); 7.1 (m, 1H); 6.97 (dd, 1H, J=9.1, 4.3 Hz); 5.3
(s, 2H); 5.1 (s, 2H).
[1345] The following compounds, Preparations 112-113, were prepared
from the appropriate starting material using analogous procedures
to Preparation II1.
Preparation II2
[1346] 2-Benzyloxy-5-iodo-benzoic acid benzyl ester.
Preparation II3
[1347] 2-Benzyloxy-5-methyl-benzoic acid benzyl ester.
Preparation JJ1
[1348] 2-Benzyloxy-5-bromo-benzyl alcohol.
[1349] Sodium borohydride (339 mg, 8.93 mmol) was added to a
solution of 2-benzyloxy-5-bromo-benzaldehyde (2.6 g, 8.93 mmol) in
anhydrous ethanol (20 mL) at 0.degree. C. The reaction was allowed
to warm to room temperature and, after 3 hours, the mixture was
concentrated and the residue was dissolved in ethyl acetate (50 mL)
and washed with water (1.times.25 mL), 1N HCl solution (1.times.25
mL) and brine (1.times.25 mL). The solution was dried
(Na.sub.2SO.sub.4), filtered and concentrated. The residue was
purified by flash chromatography (8% ethyl acetate/hexane) to
afford 2.58 g of the title compound as a colorless oil (98%).
[1350] MS 292 (M+H).sup.+. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 7.39 (m, 7H); 6.9 (d, 1H, J=8.7 Hz); 5.06 (s, 2H); 4.62 (s,
2H); 2.2 (bs, 1H).
Preparation KK1
[1351] 2-Benzyloxy-5-bromo-benzaldehyde.
[1352] Benzyl bromide (18 mL, 14.9 mmol) and cesium carbonate (8.1
g, 24.9 mmol) were added to a solution of 2-hydroxy-5-bromo
benzaldehyde 2 g, 9.95 mmol) in anhydrous DMF (25 mL) at room
temperature. The reaction was heated to 80.degree. C. for two
hours, cooled and diluted with water (50 mL). The mixture was
extracted with ethyl acetate (3.times.40 mL) and the combined
organic layers were washed with water (1.times.50 mL) and brine
(1.times.50 mL), dried (Na.sub.2SO.sub.4), filtered and
concentrated. The residue was purified by flash chromatography (10%
ethyl acetate/hexane) to afford 2.67 g of the title compound as a
colorless oil (92%). MS: 291 (M+H).sup.+. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.94 (s, 1H); 7.6 (dd, 1H, J=8.9, 2.7 Hz); 7.39
(m, 6H); 6.92 (d, 1H, J=8.9 Hz); 5.18 (s, 2H).
Preparation LL1
[1353] 2-[5-Chloro-2-(3-methylisoxazol-5-ylmethoxy)
benzyl]isoindole-1 ,3-dione
[1354] To a mixture of 2-[5-chloro-2-hydroxy
benzyl]isoindole-1,3-dione (800 mg, 2.78 mmol),
5-hydroxymethyl-3-methyl-isoxazole (373 mg, 3.3 mmol), and
triphenylphosphine (1.0 g, 4.2 mmol) in anhydrous THF (10 mL) was
added diethylazodicarboxylate (0.657 mL, 4.7 mmol). The solution
was stirred for 15 h at room temperature under anhydrous
conditions. The solvent was then removed by rotary evaporation and
the product was preadsorbed onto silica gel and purified by flash
chromatography to afford the title compound (555 mg, 52%).
[1355] C.sub.20H.sub.15ClN.sub.2O.sub.4. MW 382.81. MS 382.2
(M+H).sup.+.
[1356] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.85-7.79 (mult,
4H); 7.28 (dd, 1H, J=8.8 Hz, J=2.6 Hz); 7.18 (d, 1H, J=2.6 Hz);
7.16 (d, 1H, J=8.8 Hz); 6.39 (s, 1H); 5.25 (s, 2H); 4.67 (s, 2H);
2.17 (s, 3H).
[1357] The following compounds, Preparations LL2-LL32, were
prepared from the starting material by reaction with the
appropriate alcohol using analogous procedures to Preparation
LL1.
Preparation LL2
[1358] 5-Chloro-2-(2-morpholin4-yl-ethoxy)benzonitrile
[1359] Mp 78.0-80.0.degree. C.
[1360] C.sub.13H.sub.15ClN.sub.2O.sub.2. MW 266.73. MS 267.2
(M+H).sup.30 .
[1361] .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.55-7.45 (mult,
2H); 6.92 (d, 1H, J=8.9 Hz); 4.20 (t, 2H, J=5.6 Hz); 3.72 (t, 4H,
J=4.6 Hz); 2.86 (t, 2H, J=5.6 Hz); 2.62 (t, 4H, J=4.6 Hz).
Preparation LL3
[1362] 2-Benzyloxy-5-chlorobenzonitrile
[1363] Mp 75.0-76.5.degree. C.
[1364] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.88 (d, 1H,
J=2.7 Hz); 7.68 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.45-7.36 (mult, 4H);
7.35-7.28 (mult, 2H); 5.25 (s, 2H).
Preparation LL4
[1365] 5-Chloro-2-cyclobutylmethoxybenzonitrile
[1366] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.84 (d, 1H,
J=2.7 Hz); 7.65 (dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.45 (d, 1H, J=8.7
Hz); 4.06 (d, 2H, J=6.4 Hz); 2.75-2.63 (mult, 1H); 2.05-1.95 (mult,
2H); 1.92-1.75 (mult, 4H).
Preparation LL5
[1367] 5-Chloro-2-(3-methoxy-benzyloxy)benzonitrile
[1368] Mp 90.0-92.0.degree. C.
[1369] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.91 (d, 1H,
J=2.5 Hz); 7.70 (dd, 1H, J=8.9 Hz, J=2.5 Hz); 7.35-7.25 (mult, 2H);
7.05-6.95 (mult, 2H); 6.88 (dd, 1H, J=8.3 Hz, J=2.5 Hz); 5.25 (s,
2H); 3.73 (s, 3H).
Preparation LL6
[1370] 5-Chloro-2-(2,5-dimethoxy-benzyloxy)benzonitrile
[1371] Mp 99.0-102.0.degree. C.
[1372] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.90 (d, 1H,
J=2.7 Hz); 7.69 (dd, 1H, J=8.9 Hz, J=2.7 Hz); 7.33 (d, 1H, J=9.1
Hz); 7.02 (d, 1H, J=3.1 Hz); 6.97 (d, 1H, J=8.9 Hz); 6.87 (dd, 1H,
J=8.9 Hz, J=3.1 Hz); 5.17 (s, 2H); 3.75 (s, 3H); 3.68 (s, 3H).
Preparation LL7
[1373] 5-Chloro-2-(3-chloro-benzyloxy)benzonitrile
[1374] Mp 101.0-104.0.degree. C.
[1375] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.93 (d, 1H,
J=2.7 Hz); 7.73 (dd, 1H, J=8.9 Hz, J=2.7 Hz); 7.53 (d, 1H, J=1.5
Hz); 7.47-7.36 (mult, 2H); 7.33 (d, 1H, J=9.1 Hz); 7.25-6.95 (mult,
1 H); 5.30 (s, 2H).
Preparation LL8
[1376] 5-Chloro-2-(4-chloro-benzyloxy)benzonitrile
[1377] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.92 (d, 1H,
J=2.7 Hz); 7.71 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.46 (s, 4H); 7.33
(d, 1H, J=9.1 Hz); 5.27. (s, 2H).
Preparation LL9
[1378] 5-Chloro-2-(2-chloro-benzyloxy)benzonitrile
[1379] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.93 (d, 1H,
J=2.6 Hz); 7.73 (dd, 1H, J=9.1 Hz, J=2.6 Hz); 7.64-7.59 (mult, 1H);
7.54-7.49 (mult, 1H); 7.45-7.37 (mult, 3H); 5.32 (s, 2H).
Preparation LL10
[1380] 5-Chloro-2-(tetrahydrofuran-3-ylmethoxy)benzonitrile
[1381] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.89 (d, 1H,
J=2.7 Hz); 7.70 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.28 (d, 1H, J=9.1
Hz); 4.13-4.00 (mult, 2H); 3.80-3.70 (mult, 2H); 3.65 (dd, 1H,
J=7.7 Hz, J=6.6 Hz); 3.55-3.50 (mult, 1H); 2.70-2.60 (mult, 1H);
2.05-1.95 (mult, 1H); 1.70-1.61 (mult, 1H).
Preparation LL11
[1382] 5-Chloro-2-(4-methyl-benzyloxy)benzonitrile
[1383] Mp 106.0-108.0.degree. C.
[1384] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.89 (d, 1H,
J=2.7 Hz); 7.69 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.35-7.30 (mult, 3H);
7.19 (d, 2H, J=8.1 Hz); 5.21 (s, 2H); 2.28 (s, 3H).
Preparation LL12
[1385] 5-Chloro-2-(2-methyl-benzyloxy)benzonitrile
[1386] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.88 (d, 1H,
J=2.7 Hz); 7.70 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.39 (d, 2H, J=9.1
Hz); 7.25-7.15 (mult, 3H); 5.23 (s, 2H); 2.30 (s, 3H).
Preparation LL13
[1387] 5-Chloro-2-(3-methyl-benzyloxy)benzonitrile
[1388] .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 7.91 (d, 1H,
J=2.5 Hz); 7.71 (dd, 1H, J=9.2 Hz, J=2.5 Hz); 7.40-7.12 (mult, 5H);
5.24 (s, 2H); 2.31 (s, 3H).
Preparation LL14
[1389] 5-Chloro-2-(2-methoxy-benzyloxy)benzonitrile
[1390] Mp 114.0-115.0.degree. C.
[1391] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.89 (d, 1H,
J=2.7 Hz); 7.69 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.40 (d, 1H, J=7.5
Hz); 7.39-7.31 (mult, 2H); 7.05 (d, 1H, J=8.3 Hz); 6.97 (t, 1H,
J=7.5 Hz); 5.20 (s, 2H); 3.80 (s, 3H).
Preparation LL15
[1392] 5-Chloro-2-(furan-3-ylmethoxy)benzonitrile
[1393] Mp 72.0-74.0.degree. C.
[1394] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.89 (d, 1H,
J=2.7 Hz); 7.81 (s, 1H); 7.73-7.67 (mult, 2H); 7.38 (d, 1H, J=9.1
Hz); 6.56 (s, 1H); 5.13 (s, 2H).
Preparation LL16
[1395] 5-Chloro-2-(4-methoxy-benzyloxy)benzonitrile
[1396] Mp 84.0-85.0.degree. C.
[1397] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.88 (d, 1H,
J=2.7 Hz); 7.69 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.41-7.30 (mult, 3H);
6.98-6.92 (mult, 2H); 5.18 (s, 2H); 3.73 (s, 3H).
Preparation LL17
[1398] 5-Chloro-2-cyclopentylmethoxybenzonitrile.
[1399] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.86 (d, 1H,
J=2.7 Hz); 7.67 (dd, 1H, J=9.1 Hz, J=27 Hz); (d, 1H, J=9.1 Hz);
3.99 (d, 2H, J=6.8 Hz); 2.30 (sept, 1H, J=7.4 Hz); 1.80-1.68 (mult,
2H); 1.65-1.48 (mult, 4H); 1.38-1.26 (mult, 2H).
Preparation LL18
[1400] 3-(2-Morpholin-4-yl-ethoxy)benzaldehyde
[1401] C.sub.13H.sub.17NO.sub.3. MW 235.29. MS 236.1
(M+H).sup.+.
[1402] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 9.94 (s, 1H);
7.51-7.43 (mult, 2H); 7.42 (bs, 1H) 7.29-7.22 (mult, 1H); 4.14 (t,
2H, J=5.6 Hz); 3.55 (t, 4H, J=3.7 Hz); 2.72-2.63 (mult, 2H);
2.50-2.43 (mult, 4H).
Preparation LL19
[1403]
5-Chloro-2-[3-(2-morpholin-4-yl-ethoxy)benzyloxy]benzonitrile
[1404] C.sub.20H.sub.21ClN.sub.2O.sub.3. MW 372.86. MS 373.1
(M+H).sup.+.
[1405] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.89 (d, 1H,
J=2.7 Hz); 7.68 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.29 (d, 1H, J=9.1
Hz); 7.25 (d, 1H, J=7.7 Hz); 7.00 (bs, 1H); 6.96 (d, 1H, J=7.5 Hz);
6.87 (d, 1H, J=7.5 Hz); 5.21 (s, 2H); 4.04 (t, 2H, J=5.8 Hz);
3.57-3.49 (mult, 4H); 2.67-2.58 (mult, 2H); 2.42-2.38 (mult,
4H).
Preparation LL20
[1406] 5-Chloro-2-(tetrahydrofuran-3-ylmethoxy)benzonitrile
[1407] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.89 (d, 1H,
J=2.7 Hz); 7.70 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.28 (d, 1H, J=9.1
Hz); 4.13-4.00 (mult, 2H); 3.80-3.70 (mult, 2h); 3.65 (dd, 1H,
J=7.7 Hz, J=6.6 Hz); 3.55-3.50 (mult, 1H); 2.70-2.60 (mult, 1H);
2.05-1.95 (mult, 1H); 1.70-1.61 (mult, 1H).
Preparation LL21
[1408] 5-Chloro-2-(tetrahydrofuran-3-ylmethoxy)benzonitrile
[1409] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.89 (d, 1H,
J=2.7 Hz); 7.70 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.28 (d, 1H, J=9.1
Hz); 4.13-4.00 (mult, 2H); 3.80-3.70 (mult, 2H); 3.65 (dd, 1H,
J=7.7 Hz, J=6.6 Hz); 3.55-3.50. (mult, 1H); 2.70-2.60 (mult, 1H);
2.05-1.95 (mult, 1H); 1.70-1.61 (mult, 1H).
Preparation LL22
[1410] 5-Chloro-2-(furan-2-ylmethoxy)benzonitrile
[1411] Mp 85.0-88.0.degree. C.
[1412] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.89 (d, 1H,
J=2.7 Hz); 7.78-7.69 (mult, 2H); 7.44 (d, 1H, J=9.1 Hz); 6.64 (d,
1H, J=3.1 Hz); 6.46 (bs, 1H); 5.25 (s, 2H).
Preparation LL23
[1413]
5-Chloro-2-(2,2,7,7-tetramethyltetrahydro-bis[1,3]dioxolo[4,5-b;4',-
5'-d]pyran-5-ylmethoxy)benzonitrile
[1414] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.88 (d, 1H,
J=2.7 Hz); 7.67 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.31 (d, 1H, J=9.1
Hz); 5.44 (d, 1H, J=5.0 Hz); 4.64 (dd, 1H, J=7.9 Hz, J=1.9 Hz);
4.37-4.32 (mult, 2H); 4.30 (dd, 1H, J=10.4 Hz, J=4.2 Hz); 4.19-4.11
(mult, 1H); 4.08-4.02 (mult, 1H); 1.34 (d, 6H, J=1.5 Hz); 1.26 (d,
6H, j=10.2 Hz).
Preparation LL24
[1415] 5-Chloro-2-(2,5-dimethylfuran-3-ylmethoxy)benzonitrile
[1416] Mp 103.0-105.0.degree. C.
[1417] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.87 (d, 1H,
J=2.7 Hz); 7.70 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.33 (d, 1H, J=9.1
Hz); 6.03 (s, 1H); 5.00 (s, 2H); 2.23 (s, 3H); 2.17 (s, 3H).
Preparation LL25
[1418] 5-Chloro-2-(5-dimethylaminomethylfuran-2-ylmethoxy)
benzonitrile
[1419] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.86 (dd, 1H,
J=2.7 Hz, J=1.0 Hz); 7.69 (ddd, 1H, J=9.1 Hz, J=2.7 Hz, J=1.0 Hz);
7.40(d, 1H, J=9.1 Hz); 6.54 (d, 1H, J=3.1 Hz); 6.23 (d, 1H, J=3.1
Hz); 5.18 (s, 2H); 3.36 (s, 2H); 2.06 (d, 6H, J=1.0 Hz).
Preparation LL26
[1420]
2-[5-Chloro-2-(thiazol-2-ylmethoxy)benzyl]isoindole-1,3-dione
[1421] Mp 205.0-205.5
[1422] C.sub.19H.sub.13ClN.sub.2O.sub.3S. MW 384.84. MS 385.1
(M+H).sup.+.
[1423] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.90-7.80 (mult,
4H); 7.78 (d, 1H, J=3.1 Hz); 7.74 (d, 1H, J=3.1 Hz); 7.29 (dd, 1H,
J=8.7 Hz, J=2.5 Hz); 7.19 (d, 1H, J=2.5 Hz); 7.16 (d, 1H, J=8.7
Hz); 5.48 (s, 2H); 4.74 (s, 2H).
Preparation LL27
[1424]
2-[2-(Benzofuran-2-ylmethoxy)-5-chloro-benzyl]isoindole-1,3-dione
[1425] Mp 155.0-156.0.degree. C.
[1426] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.83-7.75 (mult,
4H); 7.59 (d, 1H, J=7.5 Hz); 7.37 (d, 1H, J=7.5 Hz); 7.33-7.17
(mult, 5H); 6.98 (s, 1H); 5.29 (s, 2H); 4.69 (s, 2H).
Preparation LL28
[1427]
2-[5-Chloro-2-(isothiazol-5-ylmethoxy)benzyl]isoindole-1,3-dione
[1428] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.48 (s, 1H);
7.89-7.79 (mult, 4H); 7.42 (s, 1H); 7.30 (dd, 1H, J=8.7 Hz, J=2.5
Hz); 7.20 (d, 1H, J=2.5 Hz); 7.14 (d, 1H, J=8.7 Hz); 5.56 (s, 2H);
4.73 (s, 2H).
Preparation LL29
[1429]
2-[5-Chloro-2-(thiophen-2-ylmethoxy)benzyl]isoindole-1,3-dione
[1430] Mp 135.0-137.0.degree. C.
[1431] C.sub.20H.sub.14ClNO.sub.3S. MW 383.86. MS 383.1
(M+H).sup.+.
[1432] .sup.1H MNR (400 MHz, DMSO-d.sub.6) .delta. 7.84-7.78 (mult,
4H); 7.47 (dd, 1H, J=5.0 Hz, J=1.2 Hz); 7.27-7.22 (mult, 1H),
7.19-7.12 (mult, 3H); 6.95 (dd, 1H, J=5.0 Hz, j==3.5 Hz); 5.31 (s,
2H); 4.65 (s, 2H).
Preparation LL30
[1433]
2-[5-Chloro-2-(quinolin-2-ylmethoxy)benzyl]isoindole-1,3-dione
[1434] Mp 190.0-192.0.degree. C.
[1435] C.sub.25H.sub.17ClN.sub.2O.sub.3. MW 428.89. MS 429.1
(M+H).sup.+.
[1436] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.20 (d, 1H,
J.=8.5 Hz); 8.06 (d, 1H, J=8.5 Hz); 7.90-7.80 (mult, 3H); 7.79-7.67
(mult, 4H); 7.56 (t, 1H, J=7.3 Hz); 7.17-7.08 (mult, 2H); 6.86 (d,
1H, J=8.5 Hz); 5.42 (s, 2H); 5.02 (s, 2H).
Preparation LL31
[1437]
2-[5-Chloro-2-(4-methyl-[1,2,3]thiadiazol-5-ylmethoxy)benzyl]isoind-
ole-1,3-dione
[1438] Mp 205.0-208.0.degree. C.
[1439] C.sub.19H.sub.14ClN.sub.3O.sub.3S. MW 399.86. MS 400.1
(M+H).sup.+.
[1440] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.82 (s, 4H);
7.36 (d, 1H, J=8.7 Hz); 7.26 (bs, 1H); 7.19 (d, 1H, J=8.7 Hz); 5.55
(s, 2H); 4.69 (s, 2H); 2.61 (s, 3H).
Preparation LL32
[1441] 5-Chloro-2-(naphthalen-1-ylmethoxy)benzonitrile
[1442] Mp 128.0-130.0.degree. C.
[1443] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.12-8.05 (mult,
1H); 7.97-7.85 (mult, 3H); 7.74 (dd, 1H, J=9.1 Hz, J=2.7 Hz); 7.68
(d, 1H, J=6.9 Hz); 7.60-7.44 (mult, 4H); 5.71 (s, 2H).
Preparation MM1
[1444]
2-[5-Chloro-2-(pyridin-3-ylmethoxy)benzyl]isoindole-1,3-dione
[1445] To a solution of 2-[5-chloro-2-hydroxy
benzyl]isoindole-1,3-dione (300 mg, 1.04 mmol), and
3-picolylchloride hydrochloride (171 mg, 1.04 mmol) in anhydrous
N,N-dimethylformamide (10 mL), cooled to 0.degree. C., was added a
60% dispersion of sodium hydride in oil (104 mg, 2.6 mmol). The
reaction was stirred at room temperature under anhydrous conditions
for 15 h. The reaction was quenched by the addition of H.sub.2O (5
mL) and then poured into a separatory funnel, and aqueous saturated
NaHCO.sub.3 solution (50 mL) was added. The aqueous layer was
extracted 3.times.50 mL with CH.sub.2Cl.sub.2. The organics were
combined and washed with saturated aqueous NaCl solution (100 mL),
dried over MgSO.sub.4, filtered, and concentrated on a rotary
evaporator triturated with ether, filtered and dried to yield the
title compound.
[1446] Mp 173.0-175.0.degree. C.
[1447] C.sub.21H.sub.15ClN.sub.2O.sub.3. MW 378.82. MS 379.1
(M+H).sup.+.
[1448] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.61 (s, 1H);
8.49 (d, 1H, J=5.0 Hz); 7.84-7.78 (mult, 5H); 7.34 (dd, 1H, J=7.9
Hz, J=4.8 Hz); 7.27 (dd, 1H, J=8.7 Hz, J=2.1 Hz); 7.17 (d, 1H,
J=1.9 Hz); 7.11 (d, 1H, J=8.7 Hz); 5.15 (s, 2H); 4.70 (s, 2H).
[1449] The following compound, preparation MM2, was prepared from
the appropriate starting material using analogous procedures to
Preparation MM1.
Preparation MM2
[1450]
2-[2-(Benzothiazol-2-ylmethoxy)-5-chlorobenzyl]isoindole-1,3-dione
[1451] Mp 209.0-211.0.degree. C.
[1452] C.sub.23H.sub.15ClN.sub.2O.sub.3S. MW434.90. MS 435.1
(M+H).sup.+.
[1453] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 8.03 (d, 1H,
J=8.0 Hz); 7.93 (d, 1H, J=8.0 Hz); 7.88-7.76 (mult, 4H); 7.49 (td,
1H, J=7.7 Hz, J=1.2 Hz); 7.41 (td, 1H, J=7.7 Hz, J=1.2 Hz); 7.29
(dd, 1H, J=8.7 Hz, J=2.7 Hz); 7.20 (d, 1H, J=2.7 Hz); 7.17 (d, 1H,
J=8.7 Hz); 5.63 (s, 2H); 4.80 (s, 2H).
Preparation NN1
[1454] 4-Methyl-[1,2,3]thiadiazol-5-yl)methanol
[1455] To a mixture of 4-methyl-[1,2,3]thiadiazole-5-carboxylic
acid methyl ester (923 mg, 5.84 mmol) in anhydrous ethanol (32 mL)
was added NaBH.sub.4 (992 mg, 26.2 mmol).
[1456] The reaction was stirred at room temperature for 15 h, under
anhydrous conditions. The reaction was then cooled to 0.degree. C.
and saturated aqueous NH.sub.4Cl (25 mL) was added. The mixture was
poured into water (25 mL), extracted with CH.sub.2Cl.sub.2
(4.times.40 mL), dried over MgSO.sub.4, and concentrated on the
rotary evaporator to an oil.
[1457] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 6.00 (bs, 1H);
4.78 (s, 2H); 2.51 (s, 3H).
Preparation OO1
[1458] 4-Methyl-[1,2,3]thiadiazole-5-carboxylic acid methyl
ester
[1459] 4-Methyl-[1,2,3]thiadiazole-5-carboxylic acid (1.0 g, 6.94
mmol), concentrated H.sub.2SO.sub.4 (866 .mu.L), and methanol (12
mL) were combined and heated to reflux for 15 h. The solvent was
reduced by a rotary evaporator and the residue was poured into 15 g
of ice. The reaction was neutralized with saturated aqueous
NaHCO.sub.3. The solution was then extracted with CH.sub.2Cl.sub.2
(4.times.35 mL). The combined organics were dried over MgSO.sub.4,
filtered, and concentrated on the rotary evaporator. The product
was preadsorbed onto silica gel and purified by flash
chromatography to afford the title compound as a yellow oil.
[1460] C.sub.5H.sub.6N.sub.2O.sub.2S. MW 158.18. MS 159.0
(M+H).sup.+.
[1461] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 3.87 (s, 3H);
2.85 (s, 3H).
Preparation PP1
[1462]
3-Deoxy-1,2-O-(1-methylethylidene)-3-[(phenylmethyl)amino]-5-O-(tri-
phenylmethyl)-.alpha.-D-ribofuranose (3 step procedure)
[1463] Step 1:
[1464] To a clean and nitrogen purged glass lined reactor, was
charged CH.sub.2Cl.sub.2 (13 gal). To the reactor was added
1,2-O-(1-methylethylidene)-.alpha.-D-xylofuranose (7.0 kg, 36.8
mol) followed by pyridine (4.5 L). The reaction was cooled to
10-15.degree. C. and then charged with trityl chloride (10.8 Kg,
38.6 moles; some exotherm noted with this addition). The reaction
mixture was then stirred at 20-25.degree. C. for 5 h and judged
complete by TLC. The crude reaction mixture was washed with a 5%
aqueous acetic acid solution (2.times.11.5 gal) and with water (22
gal). The organic layer containing
1,2-O-(1-methylethylidene)-5-O-(triphenylmethyl)-.alpha.-D-xylofuranose
was carried on to Step 2 without drying or isolation.
[1465] Step 2:
[1466] To the crude containing
1,2-O-(1-methylethylidene)-5-O-(triphenylme-
thyl)-.alpha.-D-xylofuranose in CH.sub.2Cl.sub.2 was added KBr (877
g, 7.36 mol) at 25-30.degree. C. followed by water (17 gal) and
NaHCO.sub.3(15.4 kg). The reactor was cooled to 0-5.degree. C. and
2,2,6,6,-tetramethyl-1-piperidinyloxy free radical (TEMPO) radical
(280 g) was added. With 0-5.degree. C. cooling on the jackets of
the reactor was charged with high agitation: 14 gallons of Clorox
bleach over 4 h (charge is very exothermic,the bleach is added at
such a rate to keep the internal temperature between 0-50.degree.
C.).
[1467] The reaction mixture was sampled for completion by .sup.1H
NMR (CDCl.sub.3). When the reaction was judged complete, the
mixture was diluted with CH.sub.2Cl.sub.2 (38 gal) and water (24
gal). The phases were separated and the CH.sub.2Cl.sub.2 layer was
then washed with water (2.times.30 gal) and brine (16 gal). The
organic layer was atmospherically concentrated to 10-15 gal and
used directly in Step 3.
[1468] Step 3:
[1469] To the product of Step 2 in CH.sub.2Cl.sub.2 was charged
acetic acid (1.7 L). To the reactor was added benzylamine (6 L,
56.31 mol) over 30 minutes with a cooling solution of 15.degree. C.
on the jacket of the reactor (very exothermic during initial
charge, pot kept <30.degree. C.). To the reactor was added
NaBH(OAc).sub.3 (11.6 kg, 51.5 mol) over 15 minutes. The reaction
was allowed to stir for 12 h and sampled for reaction completion by
HPLC. The reaction was diluted with CH.sub.2Cl.sub.2 (26 gal) and
2N NaOH (26 gal) at a temperature of 4-14.degree. C. over 30
minutes. The phases were separated and the organic layer extracted
with water (10 gal) and brine (10 gal). The organic layer was then
concd atmospherically to 15 gal, followed by the addition of
methanol (20 gallons) and reconcentrated atmospherically to 17 gal
with the final vapor temperature at 65.degree. C. The mixture was
cooled to 10-20.degree. C. and granulated overnight. Filtration of
the crystalline solids provided 18.4 kg of
3-deoxy-1,2-O-(1-methylethylidene)-
-3-[(phenylmethyl)amino]-5-O-(tri
phenylmethyl)-.alpha.-D-ribofuranose which was calculated to have
16% by weight methanol content (80% yield corrected for residual
solvent over three steps). The material was suitable for
transformation in the next step without further drying.
[1470] [.alpha.].sub.D: +70.75 (c 1, CH.sub.2Cl.sub.2)
[1471] m.p.=1 16.2-116.3
[1472] .sup.1NMR (400 MHz, d.sub.6-DMSO): 1.31 (s, 3), 1.47 (s, 3),
2.06 (br s, 1), 2.93 (br s, 1), 3.02 (d, 1, J=5.3, 10.4), 3.27 (d,
1, J=9), 3.61 (br d, 1, J=13.7), 3.76 (m, 1), 3.80 (br d, 1,
J=13.5), 4.65 (t, 1, J=4.2), 5.82 (d, 1, J=3.9), 7.16-7.42 (m,
20).
[1473] .sup.13C NMR (100 MHz, d.sub.6-DMSO): 26.94, 51.13, 60.60,
63.70, 77.04, 79.12, 86.24, 104.75, 111.32, 127.07, 127.43, 128.07,
128.29, 128.49, 128.78, 140.81, 144.17.
[1474] Anal. Calcd for C.sub.34H.sub.35NO.sub.4: C, 78.28; H, 6.76;
N, 2.69. Found: C, 77.89; H, 6.55; N, 2.56.
Preparation QQ1
[1475]
3-Deoxy-1,2-O-(1-methylethylidene)-3-[(4-nitrobenzoyl)amino]-.alpha-
.-D-ribofuranose (3 step procedure)
[1476] Step 1:
[1477] To a solution of
3-deoxy-1,2-O-(1-methylethylidene)-3-[(phenylmethy-
l)amino]-5-O-(triphenylmethyl)-.alpha.-D-ribofuranose (200 g, 0.383
mol) in toluene (1000 mL) was added acetic acid (30 mL) followed by
10% Pd/C (48 9). The reaction was hydrogenated on a Parr shaker for
20 h. The reaction still showed 5% starting material by HPLC, so an
additional catalyst charge was added (9.6 g) and hydrogenation
continued for an additional 20 h. The solution was filtered through
celite and the catalyst rinsed with toluene (200 mL). The solution
of crude
3-amino-3-deoxy-1,2-O-(1-methylethylidene)-5-O-(triphenylmethyl)-.alpha.--
D-ribofuranose was used directly in the next reaction.
[1478] Step 2:
[1479] To the above solution was added water (2 L) and NaHCO.sub.3
(128.8 g). While stirring well, .rho.-nitrobenzoyl chloride (71.1
g, 0.383 mol) was added portionwise over 5 min to control foaming.
After an additional 5 min, the reaction was complete by HPLC. The
phases were separated and the organic layer dried (MgSO.sub.4) and
filtered to provide crude
3-deoxy-1,2-O-(1-methylethylidene)-3-[(4-nitrobenzoyl)amino]-5-O-(triphen-
ylmethyl)-.alpha.-D-ribofuranose which was used directly in the
next step.
[1480] Step 3:
[1481] To the crude
3-deoxy-1,2-O-(1-methylethylidene)-3-[(4-nitrobenzoyl)-
amino]-5-O-(triphenylmethyl)-.alpha.-D-ribofuranose solution from
above was added methanol (890 mL) and conc. HCl (0.6 mL). The
reaction was monitored by HPLC, and after 3.5 h there was a couple
percent starting material as well as several percent of byproducts,
and the reaction was quenched by the addition of NaHCO.sub.3 (18.5
g). The solution was concentrated with the aid of vacuum to remove
all methanol; toward the end the solution was heated under
atmospheric pressure to a pot temperature of 60.degree. C. Water
(93.5 mL) was added and the solution cooled to rt and allowed to
granulate overnight. The solids were filtered off and rinsed with
toluene (200 mL) then dried under vacuum at 40-45.degree. C. to
provide 117.5 g of 3-deoxy-1,2-O-(1-methylethylidene)-
-3-[(4-nitrobenzoyl)amino]-.alpha.-D-ribofuranose which contained
8.7% ash (NaHCO.sub.3 residue). This leads to a corrected yield of
107 g (83%). This material is sufficiently pure for use in further
synthetic steps. Analytically pure material can be obtained by a
further toluene/water repulp.
[1482] [.alpha.].sub.D: +53.25 (c 1, CH.sub.2Cl.sub.2)
[1483] m.p.=187.2-187.6
[1484] .sup.1H NMR (400 MHz, CDCl.sub.3): 1.38 (s, 3), 1.58 (s, 3),
3.18 (dd, 1, J=6.6, 7,7), 3.72-3.80 (m, 1), 3.87-3.94 (m, 2), 4.43
(ddd, 5.1, 9.0, 9.0), 4.72 (dd, 1, J=4.0, 4.9), 5.93 (d, 1, J=3.8),
6.77 (d, 1, J=8.1), 7.96 (d, 2, J=8.8), 8.30 (d, 2, J=8.9).
[1485] .sup.13C NMR (75 MHz, CDCl.sub.3): 26.38, 26.51, 52.32,
60.77, 78.96, 80.41, 104.24, 112.89, 124.05, 128.40, 138.59,
149.97, 166.08.
[1486] M.S. (AP-) =337.2
[1487] Anal. Calcd for C.sub.15H.sub.18N.sub.2O.sub.7: C, 53.25; H,
5.36; N, 8.28. Found: C, 53.38; H, 5.62; N, 8.33.
Preparation RR1
[1488]
1-(3-deoxy-1,2-O-(1-methylethylidene)-3-[(4-nitrobenzoyl)amino]-.al-
pha.-D-ribofuranuronoyl]-piperidine (2 step procedure)
[1489] To a mixture of CH.sub.2Cl.sub.2 (2700 mL) and water (1800
mL) is added
3-deoxy-1,2-O-(1-methylethylidene)-3-[(4-nitrobenzoyl)amino]-.alpha-
.-D-ribofuranose (300 g, 0.887 mol), KBr (21.1 g, 0.177 mol),
NaHCO.sub.3 (408 g, 4.86 mol), and Et.sub.4NCl (12.3 g, 0.044 mol).
The solution was cooled to 0-5.degree. C. and TEMPO (6.9 g, 0.044
mol) was added. A solution of commercial Clorox bleach (4400 mL,
5.25% NaOCl) was added via addition funnel over 80 min at such a
rate that the internal temperature is maintained below 5.degree. C.
After an additional 1 h, the reaction was diluted with water (7500
mL) and warmed to rt. The organic phase was removed and discarded.
The pH of the aq. layer was adjusted to pH=2.4 by the addition of
conc. HCl (560 mL); EtOAc (4000 mL) was added, and after mixing the
phases separated. The aq. layer was extracted again with EtOAc
(2000 mL) and the combined EtOAc extracts were dried (MgSO.sub.4),
filtered, and concentrated to a thin oil (still contains residual
EtOAc). This oil was used directly in the next step.
[1490] Step 2:
[1491] To the crude
3-deoxy-1,2-O-(1-methylethylidene)-3-[(4-nitrobenzoyl)-
amino]-.alpha.-D-ribofuranuronic acid was added CH.sub.2Cl.sub.2
(3100 mL) and the solution cooled to 0-5.degree. C. To this
solution was added NEt.sub.3 (310 mL) dropwise, followed by DMF
(3.4 mL). To the mixture was slowly added oxalyl chloride (85 mL),
causing significant gas evolution. The addition rate was controlled
such that the reaction temperature does not rise above 5.degree. C.
(addition time=1 h). After an additional 15 min, piperidine (114
mL) was added dropwise to the reaction mixture, at such a rate to
maintain internal temperature below 5.degree. C. (addition time=45
min). After an additional 1 h, the reaction was diluted with water
(3 L). The phases were separated and the organic layer extracted
with aq. NaHCO.sub.3 (1.5 L water, 100 g NaHCO.sub.3). The layers
were separated and the organic layer dried (MgSO.sub.4), filtered,
and transferred to a flask equipped for distillation. The majority
of the solvent was removed by distillation (3200 mL), followed by
the addition of heptane (500 mL) and further heating until the
distillate temp. reached 58.degree. C. Additional heptane was added
(1 L) and the heating continued until the distillate reached
65.degree. C. An additional 300 mL of distillate was removed with
the aid of mild vacuum, followed by cooling and addition of
CH.sub.2Cl.sub.2 (45 mL). This solution was allowed to stir
overnight. The solids obtained were still rather sticky, so
additional CH.sub.2Cl.sub.2 (120 mL) was added and the material
stirred at rt for 3 h. The solids were filtered, and rinsed with
heptane (80 mL) to provide 242 g (65% over 2 steps)
1-[3-deoxy-1,2-O-(1-methyleth-
ylidene)-3-[(4-nitrobenzoyl)amino]-.alpha.-D-ribofuranuronoyl]-piperidine.
Estimated repulp solution: 12-14% CH.sub.2Cl.sub.2 in heptane.
[1492] [.alpha.].sub.D: +72.1 (c 1, CH.sub.2Cl.sub.2)
[1493] .sup.1H NMR (400 MHz, CDCl.sub.3): 1.36 (s, 3), 1.57 (s, 3),
1.51-1.68 (m, 6), 3.30-3.45 (m, 2), 3.60-3.74 (m, 2), 4.67 (d, 1,
J=8.7), 4.84 (dd, 1, J=3.7, 5.0), 4.90 (ddd, 1, J=5.0, 7.9, 8.3),
5.95 (d, 1, J=3.3), 6.69 (d, 1, J=7.5), 7.94 (d, 2, J=8.7), 8.26
(d, 2, J=9.1).
[1494] 13C NMR (100 MHz, CDCl.sub.3): 24.68, 25.64, 26.57, 26.74,
27.16, 43.79, 46.88, 54.76, 78.79, 105.16, 113.23, 124.00, 128.68,
139.66, 149.90, 165.17, 165.67.
Preparation SS1
[1495]
1-[1,2-di-O-acetyl-3-deoxy-3-[(4-nitrobenzoyl)amino]-D-ribofuranuro-
noyl]-piperidine -mix of anomers (2 step procedure):
[1496] Step 1:
[1497] To a solution of TFA (345 mL) and water (85 mL) was added )
1-[3-deoxy-1,2-O-(1-methylethylidene)-3-[(4-nitrobenzoyl)amino]-.alpha.-D-
-ribofuranuronoyl]-piperidine (150 g, 0.357 mol). The reaction was
held at room temperature for 5.5 h, then was. slowly added to a
quench solution of water (7.5 L) containing NaCl (2250 g),
NaHCO.sub.3 (450 g), and CH.sub.2Cl.sub.2 (3.75 L). Once the quench
was complete, the mixture was stirred an additional 15 min, and the
phases separated. The water layer was extracted with
CH.sub.2Cl.sub.2 (1.5 L) and the combined organic layers were dried
(MgSO.sub.4), filtered, and this solution used directly in the next
step.
[1498] Step 2:
[1499] To the above solution was added triethylamine (225 mL)
followed by acetic anhydride (137 mL, added dropwise to keep
temperature below 30.degree. C.). After 45 min, the reaction was
confirmed complete by HPLC analysis. To the reaction was slowly
added 2N NaOH (1 L) with water cooling to maintain temp at
30.degree. C. Once complete, the phases were separated and the
organic layer extracted with water (800 mL) and concentrated to a
dark oil. To the residue was added EtOAc (500 mL) and the solution
filtered through SiO.sub.2 (120 g) and washed with EtOAc (200 mL).
The eluents were concentrated to provide 132 g (80% over two steps)
of 1-[1,2-di-O-acetyl-3-deoxy-3-[(4-nitrobenzoyl)amino]-D-ribofura-
nuronoyl]-piperidine as a yellow foam.
[1500] Note: ratio of anomers .about.3:2. NMR data are listed for
the mixture, and for simplicity the integration numbers are
approximated as if the ratio were 1:1.
[1501] .sup.1H NMR (400 MHz, CDCl.sub.3): 1.44-1.74 (m, 12), 2.08
(s, 3), 2.09 (s, 3), 2.12 (s, 3), 2.16 (s, 3), 3.22-3.80 (m, 8),
4.84 (d, 1, J=6.8), 5.05 (s, 1), 5.11 (dd, 1, J=6.2. 7.5), 5.45 (d,
1, J=5.35), 5.52-5.60 (m, 2), 6.18 (s, 1), 6.48 (d, 1, J=7.3), 6.60
(d, 1, J=4.5), 6.96 (d, 1, J=7.5). 7.88-7.94 (m, 4), 8.26-8.36 (m,
4).
[1502] .sup.1C NMR (75 MHz, CDCl.sub.3): 20.61, 20.93, 21.22,
21.43, 24.57, 24.63, 25.64, 25.80, 26.62, 26.68, 27.15, 43.70,
43.96, 46.87, 46.98, 47.07, 52.20, 52.61, 70.51, 75.92, 78.81,
80.06, 81.51, 94.94, 98.56, 105.17, 124.01, 124.10, 124.34, 128.19,
128.54, 128.64, 139.43, 139.56, 150.00, 150.12, 165.26, 165.39,
1.65.62, 165.66, 168.59, 168.90, 169.52, 169.55.
[1503] Hi Res MS (M+H): calc: 464.1669; found: 464.1674.
[1504] Anal. Calcd for C.sub.21H.sub.25N.sub.3O.sub.9: C, 54.42; H,
5.44; N, 9.07. Found: C, 54.21; H, 5.80; N, 9.06.
Preparation TT1
[1505]
N-[[5-chloro-2-[(3-methyl-5-isoxazolyl)methoxy]phenyl]methyl]-1H-pu-
rin-6-amine (2 step procedure)
[1506] Step 1:
[1507] To a solution of
2-[[5-chloro-2-[(3-methyl-5-isoxazolyl)methoxy]phe-
nyl]methyl)-1H-isoindole-1,3 (2H)-dione (3000.0 g, 7.837 mol) in
THF (42 L) and isopropanol (31.5 L) was added 54% aqueous hydrazine
(1500 mL). The solution was. warmed to 50 .degree. C. for 5 h, upon
which the reaction was judged complete by HPLC analysis. The
solution was cooled to rt, and copious white solids were filtered
off and rinsed with THF (2500 mL). The filtrate was concentrated,
and to the oily residue was added MTBE (36 L). (Note: the
distillate containing excess hydrazine was quenched with bleach).
To the MTBE mixture was added 1N NaOH (35 L) the mixture stirred
for 10 min., and the layers separated. The organic phase was
extracted with brine (18 L), dried (Na.sub.2SO.sub.4), filtered,
and conc. to provide an oily residue. The above procedure was
repeated, and the crude
5-chloro-2-[(3-methyl-5-isoxazolyl)methoxy]-benzenemethanamine from
both runs was combined (theoretical=15.674 mol)
[1508] Step 2:
[1509] To the above combined oily residue was added isopropanol (79
L), triethylamine (4 L), and 6-chloropurine (2620 g, 16.95 mol).
The solution was heated to 75.degree. C. After .about.30 min, solid
precipitates began to form. After a total reaction time of 22 h,
the reaction was judged complete by HPLC analysis. The reaction was
cooled to rt, and to the thick slurry was added isopropanol (15 L)
to thin the solution. The slurry was filtered and the cake rinsed
with ispropanol (2.times.20 L). The solids were dried under vacuum
to provide 4700 g (81% over two steps) of
N-[[5-chloro-2-[(3-methyl-5-isoxazolyl)methoxy]phenyl]methyl]-1H-purin-
-6-amine as an off-white solid.
[1510] .sup.1H NMR (500 MHz, d.sub.6-DMSO): 2.25 (s, 3), 4.68 (br
s, 2), 5.35 (s, 2), 6.52 (br s, 1), 7.14 (br s, 1), 7.17 (d, 1,
J=8.8), 7.28 (dd, 1, J=2.0, 8.6), 8.17 (br s, 2)
[1511] .sup.13C NMR (125 MHz, d.sub.6-DMSO): 11.6, 38.6, 61.7,
105.4, 114.3, 119.3, 125.6, 127.4, 127.8, 131.2, 140.0, 150.7,
153.04, 154.4, 154.7, 160.4, 167.9.
[1512] m.p. 264.2-265.8.
[1513] Anal. Calcd for C.sub.17H.sub.15N.sub.6O.sub.2Cl: C, 55.07;
H, 4.08; N, 22.67; Cl, 9.56. Found: C, 54.98; H, 4.18; N, 22.54;
Cl, 9.74.
[1514]
1-[2-O-acetyl-1-[6-[[[5-chloro-2-[(3-methyl-5-isoxazolyl)methoxy]ph-
enyl]methyl]amino]-9H-purin-9-yl]-1,3-dideoxy-3-[(4-nitrobenzoyl)amino]-.b-
eta.-D-ribofuranuronoyl]-piperidine: To a solution of
N-[[5-chloro-2-[(3-methyl-5-isoxazolyl)methoxy]phenyl]methyl]-1H-purin-6--
amine (36.54 g, 0.0985 mol) in DME (175 mL) was added
trimethylsilyltrifluoromethanesulfonate (TMSOTf) (50.00 g, 0.2250
mol). This solution was heated to 65.degree. C. In a separate
flask,
1-[1,2-di-O-acetyl-3-deoxy-3-[(4-nitrobenzoyl)amino]-D-ribofuranuronoyl]--
piperidine (crude from previous step, 51.46 g, 0.1110 mol) was
dissolved in DME (80 mL). The
1-[1,2-di-O-acetyl-3-deoxy-3-[(4-nitrobenzoyl)amino]--
D-ribofuranuronoyl]-piperidine solution was transferred to an
addition funnel, and added to the hot
N-[[5-chloro-2-[(3-methyl-5-isoxazolyl)metho-
xy]phenyl]methyl]-1H-purin-6-amine /TMSOTf solution dropwise over
30 min. After an additional 15 min, heating was discontinued and
the reaction solution was added to CH.sub.2Cl.sub.2 (750 mL) and
sat. NaHCO.sub.3 (750 mL). The layers were separated and the aq.
layer ext. once with CH.sub.2Cl.sub.2 (200 mL). The combined
organic layers were dried (MgSO.sub.4), filtered and concentrated.
The residue was dissolved in EtOAc (250 mL), filtered, and rinsed
with EtOAc (150 mL). The EtOAc layer (400 mL total volume) was
stored at 5.degree. C. overnight, which allowed the slow formation
of crystalline pdt. The crystals were filtered and rinsed with cold
EtOAc (200 mL). The product retained a slight color, and therefore
was repulped in EtOAc (400 mL) for 1 h and refiltered, rinsing with
EtOAc (100 mL) to provide 46.76 g (54%) of the title compound as a
white solid. This material is .about.85% pure by HPLC analysis, and
was used without further purification. An analytically pure sample
was prepared by chromatography (5% IPA/CH.sub.2Cl.sub.2).
[1515] [.alpha.].sub.D: -51.8 (c 1, CH.sub.2Cl.sub.2)
[1516] .sup.1NMR (500 MHz, d.sub.6-DMSO): 1.40-1.56 (m, 6), 2.02
(s, 3), 2.25 (s, 3), 3.40-3.55 (m, 4), 4.69 (br s, 1 or 2), 5.20
(d, 1, J=5.6), 5.35 (s, 2), 5.35-5.40 (m, 1), 5.93 (app t, 1,
J=5.4), 6.52 (br s, 2), 7.15 (br s, 1), 7.17 (d, 1, J=8.8), 7.29
(dd, 1, J=2.5, 8.7), 8,10 (d, 2, J=8.7), 8.24 (s, 1), 8.37 (d, 2,
J=8.7), 8.42 (br s, 1), 8.51 (br s, 1), 9.18 (d, 1, J=7.9).
[1517] .sup.13C NMR (125 MHz, D.sub.6-DMSO): 10.94, 20.43, 23.83,
25.21, 26.14, 37.94, 42.76, 46.04, 52.42, 61.07, 74.00, 77.81,
86.33, 104.77, 113.77, 119.27, 123.64, 124.97, 126.71, 127.24,
129.05, 130.26, 139.25, 139.49, 148.82, 149.24, 152.92, 153.70,
154.43, 159.71, 165.21, 165.53, 167.22, 169.22.
[1518] M.S. (AP+)=774.2
[1519] Anal. Calcd for C.sub.36H.sub.36N.sub.9O.sub.9Cl: C, 55.85;
H, 4.69; N, 16.28; Cl, 4.58. Found: C, 55.82; H, 4.77; N, 16.28;
Cl, 4.63.
Preparation UU1
[1520]
3-amino-1-[6-[[[5-chloro-2-[(3-methyl-5-isoxazolyl)methoxy]phenyl]m-
ethyl]amino]-9H-purin-9-yl]-1,3-dideoxy-.beta.-D-ribofuranuronic
acid: To
1-[2-O-acetyl-1-[6-[[[5-chloro-2-[(3-methyl-5-isoxazolyl)methoxy]phenyl]m-
ethyl]amino]-9H-purin-9-yl]-1,3-dideoxy-3-[(4-nitrobenzoyl)amino]-.beta.-D-
-ribofuranuronoyl]-piperidine (250.0 g, 0.3229 mol, 88% purity) was
added THF (1250 mL, water (650 mL), MeOH (350 mL), and solid KOH
(85%, 127.9 g). The solution was heated to 65.degree. C. for 23 h
and allowed to cool to rt. To the solution was added water (1250
mL) and MTBE (1250 mL).
[1521] The phases were separated and the MTBE layer discarded. To
the aqueous layer was added conc. HCl to adjust to pH=5.6, forming
significant precipitate. After granulating for 2 h, the slurry was
filtered. The wet cake was added back to a flask with water (200
mL) and THF (1800 mL) and stirred 1 h. The slurry was refiltered,
and the wet cake again added to a flask with water (1 L). To this
slurry was added conc. HCl to pH=1.2, THF (2 L) and EtOAc (1 L)
were added, followed by additional THF to aid separation (1L). Due
to emulsion, an additional portion of THF was added (500 mL) and
the bilayer filtered through celite. The phases were then
separated, and the organic phase extracted with water (2.times.200
mL). All aqueous phases were then combined and 6M NaOH (.about.100
mL) was added to adjust pH to 5.0. The solids were filtered and
rinsed with a solution of 10/90 water/THF (1300 mL total). The
solids were dried in a vacuum oven to provide 91.02 g (55%) of the
title compound as an off-white powder.
[1522] Hi. Res. M.S.=516.1390 (M+H). Calculated =516.1398.
[1523] .sup.1H NMR (500 MHz, d.sub.6-DMSO): 2.24 (s, 3), 3.82 (br
s, 1), 4.34 (br d, 1, J=5.1), 4.66 (brs, 2), 5.34 (s, 2), 6.14 (d,
1, J=2.9), 6.52 (s, 1), 7.11 (brs, 1), 7.15 (d, 1, J=8.8), 7.27
(dd, 1, J=2.3, 8.7), 8.20 (s, 1), 8.34 (br s, 1), 8.88 (br s,
1).
[1524] .sup.13C NMR (125 MHz, d.sub.6-DMSO): 10.94, 37.88, 55.31,
61.05, 73.82, 81.57, 88.61, 104.76, 113.73, 119.34, 124.95, 126.61,
127.16, 130.39, 139.97, 148.70, 152.49, 153.67, 154.33, 159.11,
167.20, 172.06.
Preparation Vv1
[1525]
3-amino-1-[6-[[[5-chloro-2-[(3-methyl-5-isoxazolyl)methoxy]phenyl]m-
ethyl]amino]-9H-purin-9-yl]-1,3-dideoxy-N-methyl-.beta.-D-ribofuranuronami-
de: A solution of HCl/methanol was prepared by adding acetyl
chloride (8.5 mL, 0.12 mol) slowly to methanol (1 L). After 10
minutes,
3-amino-1-[6-[[[5-chloro-2-[(3-methyl-5-isoxazolyl)methoxy]phenyl]methyl]-
amino]-9H-purin-9-yl]-1,3-dideoxy-.beta.-D-ribofuranuronic acid
(40.03 g, 0.0776 mol) was added to the HCl/MeOH soln with a
methanol rinse (100 mL). The solution was heated at 50.degree. C.
for 15 h, upon which conversion of the acid to the methyl ester was
confirmed by HPLC. The solution was cooled to 37.degree. C.
followed by the addition of methylamine (600 mL of a 2.0M soln in
methanol). The reaction was reheated to 50.degree. C. for 5.5 h,
and conversion of the ester to methylamide was confirmed by HPLC.
The reaction was set up for distillation, and 250 mL of solvent was
removed by maintaining the pot temperature between 45-50.degree. C.
and using slight vacuum. The solution temperature was then raised
to 65.degree. C. at atmospheric pressure, and Darco (6.0 g) and
water (10 mL) were added. After 5 min, the solution was filtered
through celite while hot, rinsing with methanol (100 mL). The
filtrate was allowed to cool to rt while stirring overnight,
producing a crystalline product. The crystalline product was
filtered and rinsed with methanol (200 mL) to provide 30.73 g (72%)
the title compound as the hydrate. HPLC analysis of this material
showed greater than 98% purity.
[1526] It should be understood that the invention is not limited to
the particular embodiments described herein, but that various
changes and modifications may be made without departing from the
spirit and scope of this novel concept as defined by the following
claims.
* * * * *