U.S. patent application number 14/109922 was filed with the patent office on 2014-06-19 for tricyclic compounds.
This patent application is currently assigned to NOVARTIS AG. The applicant listed for this patent is Mahbub AHMED, Alexander ASHALL-KELLY, Louisa GUERITZ, Jeffrey MCKENNA, Joseph MCKENNA, Simon MUTTON, Rakesh PARMAR, Jon SHEPHERD, Paul WRIGHT. Invention is credited to Mahbub AHMED, Alexander ASHALL-KELLY, Louisa GUERITZ, Jeffrey MCKENNA, Joseph MCKENNA, Simon MUTTON, Rakesh PARMAR, Jon SHEPHERD, Paul WRIGHT.
Application Number | 20140171417 14/109922 |
Document ID | / |
Family ID | 50030372 |
Filed Date | 2014-06-19 |
United States Patent
Application |
20140171417 |
Kind Code |
A1 |
AHMED; Mahbub ; et
al. |
June 19, 2014 |
TRICYCLIC COMPOUNDS
Abstract
The present invention provides a compound of formula I or a
pharmaceutically acceptable salt thereof; ##STR00001## and its
therapeutic uses. The present invention further provides a
combination of pharmacologically active agents and a pharmaceutical
composition.
Inventors: |
AHMED; Mahbub; (Horsham,
GB) ; ASHALL-KELLY; Alexander; (London, GB) ;
GUERITZ; Louisa; (London, GB) ; MCKENNA; Jeffrey;
(Horsham, GB) ; MCKENNA; Joseph; (Hove, GB)
; MUTTON; Simon; (Guildford, GB) ; PARMAR;
Rakesh; (Hove, GB) ; SHEPHERD; Jon; (Horsham,
GB) ; WRIGHT; Paul; (Ashtead, GB) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AHMED; Mahbub
ASHALL-KELLY; Alexander
GUERITZ; Louisa
MCKENNA; Jeffrey
MCKENNA; Joseph
MUTTON; Simon
PARMAR; Rakesh
SHEPHERD; Jon
WRIGHT; Paul |
Horsham
London
London
Horsham
Hove
Guildford
Hove
Horsham
Ashtead |
|
GB
GB
GB
GB
GB
GB
GB
GB
GB |
|
|
Assignee: |
NOVARTIS AG
Basel
CH
|
Family ID: |
50030372 |
Appl. No.: |
14/109922 |
Filed: |
December 17, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61906141 |
Nov 19, 2013 |
|
|
|
61739335 |
Dec 19, 2012 |
|
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|
Current U.S.
Class: |
514/214.02 ;
514/224.5; 514/248; 514/267; 540/578; 544/234; 544/250; 544/34 |
Current CPC
Class: |
A61K 45/06 20130101;
A61K 31/55 20130101; A61P 13/12 20180101; A61K 31/519 20130101;
C07D 487/22 20130101; C07D 513/14 20130101; A61K 31/542 20130101;
C07D 487/14 20130101; C07D 495/14 20130101; C07D 487/04 20130101;
C07D 513/00 20130101; A61P 43/00 20180101; A61P 1/12 20180101; C07D
471/14 20130101 |
Class at
Publication: |
514/214.02 ;
544/34; 514/224.5; 544/250; 514/267; 544/234; 514/248; 540/578 |
International
Class: |
C07D 513/00 20060101
C07D513/00; C07D 471/14 20060101 C07D471/14; A61K 45/06 20060101
A61K045/06; C07D 487/14 20060101 C07D487/14; A61K 31/55 20060101
A61K031/55; A61K 31/542 20060101 A61K031/542; A61K 31/519 20060101
A61K031/519 |
Claims
1. A compound of the formula (I) or a pharmaceutically acceptable
salt thereof: ##STR00201## wherein R.sup.1 represents phenyl,
(C.sub.4-C.sub.7)cycloalkenyl or Het.sup.1, which R.sup.1 group may
be unsubstituted or substituted on one or two carbon atoms by
substituents R.sup.a, and may further be substituted on a nitrogen
atom with a substituent R.sup.a1; each R.sup.a independently
represents (C.sub.1-C.sub.4)alkyl, halo,
halo(C.sub.1-C.sub.4)alkyl, cyano, hydroxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy, halo(C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl, R.sup.6OC(O)--, or
R.sup.6OC(O)(C.sub.1-C.sub.4)alkyl-; R.sup.a1 represents
(C.sub.1-C.sub.4)alkyl, hydroxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-,
aryl(C.sub.1-C.sub.4)alkyl or R.sup.6OC(O)(C.sub.1-C.sub.4)alkyl-;
R.sup.2 represents (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.7)cycloalkyl, (C.sub.4-C.sub.7)cycloalkenyl, phenyl,
furanyl, thiazolyl or thienyl, which R.sup.2 may be unsubstituted
or substituted on from one to three carbon atoms with substituents
R.sup.b; each R.sup.b independently represents
(C.sub.1-C.sub.4)alkyl, halo, halo(C.sub.1-C.sub.4)alkyl, cyano,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-,
(R.sup.6).sub.2NC(O)(C.sub.1-C.sub.4)alkyl- or
R.sup.6OC(O)(C.sub.1-C.sub.4)alkyl-; X represents S, Z represents
CHR.sup.4a and n represents 1; or X represents CHR.sup.4b, Z
represents NR.sup.5 and n represents 1 or 2; or X represents
CHR.sup.4b, Z represents CHR.sup.4a and n represents 0, 1 or 2; or
X represents C(.dbd.CH.sub.2), CF.sub.2 or C(CH.sub.3).sub.2, Z
represents CHR.sup.4a and n represents 0 or 1; each R.sup.3
independently represents methyl or ethyl; when X represents S,
R.sup.4 represents hydrogen, (C.sub.1-C.sub.4)alkyl,
halo(C.sub.1-C.sub.4)alkyl, hydroxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl-,
amino(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-, phenyl,
Het.sup.1(C.sub.1-C.sub.4)alkyl-, Het.sup.2(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylS(O).sub.2NH(C.sub.1-C.sub.4)alkyl-, or
R.sup.7C(O)NH(C.sub.1-C.sub.4)alkyl-; when X represents CHR.sup.4b,
each R.sup.4 independently represents hydrogen, hydroxy,
(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy,
halo(C.sub.1-C.sub.4)alkyl, hydroxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl-,
amino(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-, amino,
(C.sub.1-C.sub.4)alkylamino-, di[(C.sub.1-C.sub.4)alkyl]amino-,
phenyl, Het.sup.1(C.sub.1-C.sub.4)alkyl-,
Het.sup.2(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylS(O).sub.2NH(C.sub.1-C.sub.4)alkyl-, or
R.sup.7C(O)NH(C.sub.1-C.sub.4)alkyl-; R.sup.4a represents hydrogen,
(C.sub.1-C.sub.4)alkyl, hydroxy(C.sub.1-C.sub.4)alkyl,
halo(C.sub.1-C.sub.4)alkyl, amino(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-,
Het.sup.1(C.sub.1-C.sub.4)alkyl-, Het.sup.2(C.sub.1-C.sub.4)alkyl-,
or R.sup.6OC(O)--; R.sup.4b represents hydrogen or methyl; R.sup.5
represents hydrogen or (C.sub.1-C.sub.4)alkyl; R.sup.6 represents
hydrogen or (C.sub.1-C.sub.4)alkyl; R.sup.7 represents
(C.sub.1-C.sub.2)alkyl, (C.sub.1-C.sub.2)alkoxy,
(C.sub.1-C.sub.2)alkoxy(C.sub.1-C.sub.2)alkyl or phenyl; Het.sup.1
represents a 5- or 6-membered heteroaryl ring comprising a) one
oxygen or sulphur atom and optionally one or two nitrogen atoms; or
b) from one to four nitrogen atoms; and Het.sup.2 represents a 4-
to 7-membered heterocyclic ring comprising a) 1 or 2 heteroatoms
selected from N, O and S; or b) --C(O)-- and 1 or 2 heteroatoms
selected from N and O.
2. A compound according to claim 1, wherein R.sup.1 represents
phenyl, cyclohexenyl, thiazolyl, pyrazolyl, thienyl, pyrimidin-2-yl
or pyridine-2-yl and wherein R.sup.1 may be unsubstituted or
substituted on one or two carbon atoms by substituents R.sup.a, and
may further be substituted on a nitrogen atom with a substituent
R.sup.a1.
3. A compound according to claim 1, wherein R.sup.2 represents
phenyl, furanyl, thiazolyl, or thieny, which R.sup.2 may be
unsubstituted or substituted on from one to three carbon atoms with
substituents R.sup.b.
4. A compound according to claim 1, wherein each R.sup.b
independently represents (C.sub.1-C.sub.2)alkyl, halo,
halo(C.sub.1-C.sub.2)alkyl, hydroxy(C.sub.1-C.sub.2)alkyl, or
cyano.
5. A compound according to claim 1, wherein each R.sup.3 represents
methyl.
6. A compound according to claim 1, wherein R.sup.4 represents
hydrogen, methyl, phenyl or HOCH.sub.2--.
7. A compound according to claim 1, wherein X represents S, Z
represents CHR.sup.4a and n represents 1.
8. A compound according to claim 1, wherein R.sup.4a represents
hydrogen or methyl.
9. A compound according to claim 1, wherein the compound is of
formula (Ia): ##STR00202##
10. A compound of formula (I) which is selected from:
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[4',-
5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[4-
',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',-
5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(3-Chlorophenyl)-1,3-dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',5':3-
,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
1,3-Dimethyl-5-phenyl-10-(m-tolyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrr-
olo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(m-tolyl)-7,8-dihydro-1H-pyrimido[-
4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-(m-tolyl)-7,8-dihydro-1H-pyrimid-
o[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
1,3-Dimethyl-10-(5-methylthiophen-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[-
4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
1,3-Dimethyl-10-(4-methylthiophen-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[-
4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',-
5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[4-
',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[4',-
5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(3-Chlorophenyl)-1,3-dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',5':3-
,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(m-tolyl)-7,8-dihydro-1H-pyrimido[-
4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
7-((dimethylamino)methyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7-
,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-d-
ione;
10-(5-chlorofuran-2-yl)-1,3,8-trimethyl-5-phenyl-7,8-dihydro-1H-pyri-
mido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
1,3,8-trimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimid-
o[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
3-(10-(5-Chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-hexahydro--
1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)benzonitrile;
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8-dihydro-
-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-5-(cyclohex-1-en-1-yl)-1,3-dimethyl-7,8-dihydro-1-
H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-5-(3,5-difluorophenyl)-1,3-dimethyl-7,8-dihydro-1-
H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8-dihydro-
-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-chlorofuran-2-yl)-5-(cyclohex-1-en-1-yl)-1,3-dimethyl-7,8-dihydro-1-
H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-1,3-diethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',5-
':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
3-Chloro-5-(10-(5-chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-h-
exahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)benzonitril-
e;
10-(2,3-Difluorophenyl)-1,3-dimethyl-5-phenyl-7,8,9,10-tetrahydropyrimi-
do[4,5-a]indolizine-2,4(1H,3H)-dione;
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimido-
[4,5-a]indolizine-2,4(1H,3H)-dione;
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimi-
do[4,5-a]indolizine-2,4(1H,3H)-dione;
5-(3-Fluorophenyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetrahyd-
ro pyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
5-(3-fluorophenyl)-1,3,9-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetra-
hydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
5-(3-fluorophenyl)-1,3,8-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetra-
hydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
5-(3-fluorophenyl)-1,3,9-trimethyl-10-(4-methylthiazol-2-yl)-7,8,9,10-tet-
rahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-m-tolyl-7,8,9,10-tetrahydropyrim-
ido[4,5-a]indolizine-2,4(1H,3H)-dione;
5-(3-Chlorophenyl)-10-(4-chlorothiazol-2-yl)-1,3-dimethyl-7,8,9,10-tetrah-
ydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
3-(1,3-Dimethyl-10-(4-methylthiazol-2-yl)-2,4-dioxo-1,2,3,4,7,8,9,10-octa-
hydro pyrimido[4,5-a]indolizin-5-yl)benzonitrile;
3-(1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4,7,8,9,10-octahydropyrimido[4,5-
-a]indolizin-10-yl)benzonitrile;
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimi-
do[4',5':3,4]pyrrolo[1,2-b]pyridazine-2,4(1H,3H)-dione;
5-(3-Chlorophenyl)-1,3-dimethyl-10-(4-methylthiazol-2-yl)-7,8,9,10-tetrah-
ydropyrimido[4',5':3,4]pyrrolo[1,2-b]pyridazine-2,4(1H,3H)-dione;
1,3-dimethyl-9-(5-methylfuran-2-yl)-5-phenyl-8,9-dihydro-1H-pyrimido[4,5--
a]pyrrolizine-2,4(3H,7H)-dione;
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-phenyl-7,8-dihyd-
ro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-chlorofuran-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimethyl-
-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-
-dione;
3-(10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-diox-
o-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin--
5-yl)benzonitrile;
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3-
,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)-
benzonitrile
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10-
H)-dione;
3-(10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-1,2,3,4,7,8,-
9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile;
10-(4-Chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl--
7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
1,3-Dimethyl-5,10-bis(4-methylthiazol-2-yl)-7,8,9,10-tetrahydropyrimido[4-
,5-a]indolizine-2,4(1H,3H)-dione;
10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8,9,10--
tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(2-methylthiazol-4-yl)-7,8,9,10--
tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
10-(5-chlorofuran-2-yl)-8-((dimethylamino)
methyl)-1,3-dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2-
,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one;
10-(4-Chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimet-
hyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,1-
0H)-dione;
10-(5-Chlorofuran-2-yl)-5-(3-ethoxyphenyl)-1,3-dimethyl-7,8-dih-
ydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
5-(3-Methoxyphenyl)-1,3-dimethyl-10-(4-methylthiazol-2-yl)-7,8-dihydro-1H-
-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-5-(3-methoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-p-
yrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(2-Chlorothiazol-4-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8-d-
ihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione-
;
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8-di-
hydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-imidazol-4-yl)-7,8-di-
hydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-4-yl)-7,8-dih-
ydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(5-methyl-1,3,4-oxadiazol-2-yl)-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one;
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(4-methyloxazol-2-yl)-7,8-dihy-
dro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8-dih-
ydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-chlorofuran-2-yl)-5-(3-ethoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-py-
rimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-5-(3-methoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-p-
yrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(5-methyl-1,3,4-oxadiazol-2-yl)-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one;
10-(5-chlorofuran-2-yl)-5-(4-(1-hydroxyethyl)thiazol-2-yl)-1,3-dimeth-
yl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10-
H)-dione;
10-(4-chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3--
dimethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4-
(3H,10H)-dione;
9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9-dihydro-1H-pyrimido[4,-
5-a]pyrrolizine-2,4(3H,7H)-dione;
1,3-Dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyrimido[4,-
5-a]pyrrolizine-2,4(3H,7H)-dione;
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(4-methylthiazol-
-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione;
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(1-methyl-1H-pyr-
azol-3-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2-
,4(3H,10H)-dione;
10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(2-methylthiazol-
-4-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione;
9,9-Difluoro-5-(3-fluorophenyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-7,8,-
9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
8-(aminomethyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-
-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,-
10H)-dione;
3-(9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9-hexahydro--
1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile;
9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8,9-dihydr-
o-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydro-1H-p-
yrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
9-(4-Chlorothiazol-2-yl)-1,3,8,8-tetramethyl-5-phenyl-8,9-dihydro-1H-pyri-
mido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3,8,8-tetramethyl-8,9-dihyd-
ro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
9-(4-Chlorothiazol-2-yl)-1,3,8,8-tetramethyl-5-phenyl-8,9-dihydro-1H-pyri-
mido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
3-(1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-2,4-dioxo-2,3,4,7,8,9-hex-
ahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile;
5-(3-Fluorophenyl)-1,3,8,8-tetramethyl-9-(4-methylthiazol-2-yl)-8,9-dihyd-
ro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyri-
mido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
9-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3,8,8-tetramethyl-8,9-dihyd-
ro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
5-(3-Fluorophenyl)-1,3,8,8-tetramethyl-9-(4-methylthiazol-2-yl)-8,9-dihyd-
ro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
3-(1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-2,4-dioxo-2,3,4,7,8,9-hex-
ahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile;
1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyri-
mido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8,9-
,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-1,2-
,3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile;
7-((2H-1,2,3-triazol-2-yl)methyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-5--
phenyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
2-(10-(5-Chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-hexahydro--
1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)thiazole-4-carboxyli-
c acid;
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)--
2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]-
thiazin-8)methyl)methanesulfonamide;
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)-2-methoxyacetamide; Methyl
((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dioxo-
-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-8-
-yl)methyl) carbamate;
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)acetamide;
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)benzamide;
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-7-(hydroxymethyl)-1,3-dimethy-
l-8,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-5-phenyl-8,9-dihy-
dro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
7-(hydroxymethyl)-1,3-dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihy-
dro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
3-(9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3,-
4,7,8,9-hexahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile;
8-((1H-imidazol-1-yl)methyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-me-
thylthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thi-
azine-2,4(3H,10H)-dione;
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8-methylene-8,9--
dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxoox-
azolidin-3-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazine-2,4(3H,10H)-dione;
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxo
pyrrolidin-1-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][-
1,4]thiazine-2,4(3H,10H)-dione;
3-(11-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9,10,11-oct-
ahydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepin-5-yl)benzonitrile;
11-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9,10,11-tetra-
hydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione;
1,3-Dimethyl-11-(4-methylthiazol-2-yl)-5-phenyl-8,9,10,11-tetrahydro-1H-p-
yrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione;
11-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9,10,11-tetrahydro-1H-p-
yrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione; and the
pharmaceutically acceptable salts thereof.
11. A pharmaceutical composition comprising a therapeutically
effective amount of a compound according to claim 1 or a
pharmaceutically acceptable salt thereof and one or more
pharmaceutically acceptable carriers.
12. A combination comprising a therapeutically effective amount of
a compound according to claim 1 or a pharmaceutically acceptable
salt thereof and one or more therapeutically active co-agents.
13. A method of modulating CFTR activity in a subject, wherein the
method comprises administering to the subject a therapeutically
effective amount of the compound according to claim 1 or a
pharmaceutically acceptable salt thereof.
14. A method of treating a disorder or disease selected from
polycystic kidney disease and diarrhea, comprising administering to
the subject a therapeutically effective amount of the compound
according to claim 1 or a pharmaceutically acceptable salt thereof.
Description
FIELD OF THE INVENTION
[0001] The invention provides tricyclic compounds, the use thereof
for inhibiting the CFTR channel and methods of treating disease
using same.
BACKGROUND OF THE INVENTION
[0002] The cystic fibrosis transmembrane conductance regulator
protein (CFTR) is a cAMP-activated chloride channel which is
expressed in epithelial cells in mammalian airways, intestine,
pancreas, and testis (Sheppard et al., Physiol. Rev. 79:S23-45
(1999); Gadsby et al., Nature 40:477-83 (2006)). The CFTR chloride
channel is known to be associated with a number of diseases and
conditions, including cystic fibrosis (CF), polycystic kidney
disease and secretory diarrhea.
[0003] Diarrheal disease remains an area of high unmet medical
need, resulting in approximately 2 million deaths in 2002, of which
more than 95% were children under the age of 5 years. Infectious
secretory diarrhea, the result of poor sanitation and close living
conditions, is responsible for most acute episodes and there is a
defined need for an adjunct therapy to be used in combination with
existing oral rehydration and antibiotic therapies.
[0004] CFTR inhibitors are discussed by Thiagarajah and Verkman in
Clinical Pharmacology and Therapeutics (2012): 92, 3, 287-290.
SUMMARY OF THE INVENTION
[0005] There is a need to provide new CFTR inhibitors that are good
drug candidates. In particular, compounds of the invention should
bind potently to the cystic fibrosis transmembrane conductance
regulator protein whilst showing little affinity for other
receptors and show functional activity as CFTR inhibitors. They
should be well absorbed from the gastrointestinal tract, be
metabolically stable and possess favourable pharmacokinetic
properties. Furthermore, the ideal drug candidate will exist in a
physical form that is stable, non-hygroscopic and easily
formulated.
[0006] The compounds of the invention are high affinity inhibitors
of the CFTR channel and are therefore potentially useful in the
treatment of a wide range of disorders, particularly polycystic
kidney disease and diarrhea (including infectious secretory
diarrhea, travellers diarrhea, diarrhea associated with HIV and
diarrhea predominant irritable bowel syndrome (IBS)).
[0007] The treatment of polycystic kidney disease and diarrhea is a
contemplated use. All forms of polycystic kidney disease and
diarrhea are potentially treatable with the compounds of the
present invention including infectious secretory diarrhea,
travellers diarrhea, diarrhea associated with HIV and diarrhea
predominant irritable bowel syndrome (IBS).
[0008] The invention therefore provides, as Embodiment 1, a
compound of the formula (I) or a pharmaceutically acceptable salt
thereof:
##STR00002##
[0009] wherein R.sup.1 represents phenyl,
(C.sub.4-C.sub.7)cycloalkenyl or Het.sup.1, which R.sup.1 group may
be unsubstituted or substituted on one or two carbon atoms by
substituents R.sup.a, and may further be substituted on a nitrogen
atom with a substituent R.sup.a1;
[0010] each R.sup.a independently represents
(C.sub.1-C.sub.4)alkyl, halo, halo(C.sub.1-C.sub.4)alkyl, cyano,
hydroxy(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy,
halo(C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl, R.sup.6OC(O)--, or
R.sup.6OC(O)(C.sub.1-C.sub.4)alkyl-;
[0011] R.sup.a1 represents (C.sub.1-C.sub.4)alkyl,
hydroxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-,
aryl(C.sub.1-C.sub.4)alkyl or
R.sup.6OC(O)(C.sub.1-C.sub.4)alkyl-;
[0012] R.sup.2 represents (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.7)cycloalkyl, (C.sub.4-C.sub.7)cycloalkenyl, phenyl,
furanyl, thiazolyl or thienyl, which R.sup.2 may be unsubstituted
or substituted on from one to three carbon atoms with substituents
R.sup.b;
[0013] each R.sup.b independently represents
(C.sub.1-C.sub.4)alkyl, halo, halo(C.sub.1-C.sub.4)alkyl, cyano,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-,
(R.sup.6).sub.2NC(O)(C.sub.1-C.sub.4)alkyl- or
R.sup.6OC(O)(C.sub.1-C.sub.4)alkyl-;
[0014] X represents S, Z represents CHR.sup.4a and n represents 1;
or
[0015] X represents CHR.sup.4b, Z represents NR.sup.5 and n
represents 1 or 2; or
[0016] X represents CHR.sup.4b, Z represents CHR.sup.4a and n
represents 0, 1 or 2; or
[0017] X represents C(.dbd.CH.sub.2), CF.sub.2 or
C(CH.sub.3).sub.2, Z represents CHR.sup.4a and n represents 0 or
1;
[0018] each R.sup.3 independently represents methyl or ethyl;
[0019] when X represents S, R.sup.4 represents hydrogen,
(C.sub.1-C.sub.4)alkyl, halo(C.sub.1-C.sub.4)alkyl,
hydroxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl-,
amino(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-, phenyl,
Het.sup.1(C.sub.1-C.sub.4)alkyl-, Het.sup.2(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylS(O).sub.2NH(C.sub.1-C.sub.4)alkyl-, or
R.sup.7C(O)NH(C.sub.1-C.sub.4)alkyl-;
[0020] when X represents CHR.sup.4b, each R.sup.4 independently
represents hydrogen, hydroxy, (C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy, halo(C.sub.1-C.sub.4)alkyl,
hydroxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl-,
amino(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-, amino,
(C.sub.1-C.sub.4)alkylamino-, di[(C.sub.1-C.sub.4)alkyl]amino-,
phenyl, Het.sup.1(C.sub.1-C.sub.4)alkyl-,
Het.sup.2(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylS(O).sub.2NH(C.sub.1-C.sub.4)alkyl-, or
R.sup.7C(O)NH(C.sub.1-C.sub.4)alkyl-;
[0021] R.sup.4a represents hydrogen, (C.sub.1-C.sub.4)alkyl,
hydroxy(C.sub.1-C.sub.4)alkyl, halo(C.sub.1-C.sub.4)alkyl,
amino(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-,
Het.sup.1(C.sub.1-C.sub.4)alkyl-, Het.sup.2(C.sub.1-C.sub.4)alkyl-,
or R.sup.6OC(O)--;
[0022] R.sup.4b represents hydrogen or methyl;
[0023] R.sup.5 represents hydrogen or (C.sub.1-C.sub.4)alkyl;
[0024] R.sup.6 represents hydrogen or (C.sub.1-C.sub.4)alkyl;
[0025] R.sup.7 represents (C.sub.1-C.sub.2)alkyl,
(C.sub.1-C.sub.2)alkoxy,
(C.sub.1-C.sub.2)alkoxy(C.sub.1-C.sub.2)alkyl or phenyl;
[0026] Het.sup.1 represents a 5- or 6-membered heteroaryl ring
comprising a) one oxygen or sulphur atom and optionally one or two
nitrogen atoms; or b) from one to four nitrogen atoms; and
[0027] Het.sup.2 represents a 4- to 7-membered heterocyclic ring
comprising a) 1 or 2 heteroatoms selected from N, O and S; or b)
--C(O)-- and 1 or 2 heteroatoms selected from N and O.
[0028] In another embodiment, the invention provides a
pharmaceutical composition comprising a therapeutically effective
amount of a compound according to the definition of formula (I), or
a pharmaceutically acceptable salt thereof, or subformulae thereof
and one or more pharmaceutically acceptable carriers.
[0029] In another embodiment, the invention provides a combination,
in particular a pharmaceutical combination, comprising a
therapeutically effective amount of the compound according to the
definition of formula (I), or a pharmaceutically acceptable salt
thereof, or subformulae thereof and one or more therapeutically
active agent.
[0030] In another embodiment, the invention provides a method of
modulating CFTR activity in a subject, wherein the method comprises
administering to the subject a therapeutically effective amount of
a compound according to the definition of formula (I), or a
pharmaceutically acceptable salt thereof, or subformulae
thereof.
[0031] In another embodiment, the invention provides a method of
treating a disorder or disease selected from polycystic kidney
disease and diarrhea, comprising administering to the subject a
therapeutically effective amount of a compound according to the
definition of formula (I), or a pharmaceutically acceptable salt
thereof, or subformulae thereof.
DETAILED DESCRIPTION
[0032] The invention therefore provides a compound of the formula
(I) as described hereinabove as Embodiment 1.
[0033] Unless specified otherwise, the term "compounds of the
present invention" refers to compounds of formula (I) and formula
(Ia), salts of the compound, hydrates or solvates of the compounds,
as well as all stereoisomers (including diastereoisomers and
enantiomers), tautomers and isotopically labeled compounds
(including deuterium substitutions). Compounds of the present
invention further comprise polymorphs of compounds of formula I and
formula (Ia) and salts thereof.
[0034] For purposes of interpreting this specification, the
following definitions will apply and whenever appropriate, terms
used in the singular will also include the plural and vice
versa.
[0035] As used herein, the term "C.sub.1-6alkyl" refers to a fully
saturated branched or unbranched hydrocarbon moiety having 1 to 6
carbon atoms. The terms "C.sub.1-4alkyl" and "C.sub.1-2alkyl" are
to be construed accordingly. Representative examples of
C.sub.1-6alkyl include, but are not limited to, methyl, ethyl,
n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl,
n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl,
2,2-dimethylpentyl, 2,3-dimethylpentyl, and n-heptyl.
[0036] As used herein, the term "haloC.sub.1-6alkyl" refers to a
C.sub.1-6alkyl group as defined herein, wherein at least one of the
hydrogen atoms is replaced by a halo atom. The haloC.sub.1-6alkyl
group can be monohaloC.sub.1-6alkyl, dihaloC.sub.1-6alkyl or
polyhaloC.sub.1-6alkyl including perhaloC.sub.1-6alkyl. A
monohaloC.sub.1-6alkyl can have one iodo, bromo, chloro or fluoro
within the alkyl group. DihaloC.sub.1-6alkyl and
polyhaloC.sub.1-6alkyl groups can have two or more of the same halo
atoms or a combination of different halo groups within the alkyl.
Typically the polyhaloC.sub.1-6alkyl group contains up to 12, or
10, or 8, or 6, or 4, or 3, or 2 halo groups.
[0037] Non-limiting examples of haloC.sub.1-6alkyl include
fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl,
dichloromethyl, trichloromethyl, pentafluoroethyl,
heptafluoropropyl, difluorochloromethyl, dichlorofluoromethyl,
difluoroethyl, difluoropropyl, dichloroethyl and dichloropropyl. A
perhaloC.sub.1-6alkyl group refers to an C.sub.1-6alkyl group
having all hydrogen atoms replaced with halo atoms.
[0038] The term "aryl" refers to an aromatic hydrocarbon group
having 6-20 carbon atoms in the ring portion. Typically, aryl is
monocyclic, bicyclic or tricyclic aryl having 6-20 carbon atoms and
includes one or more aromatic rings fused to one or more
non-aromatic hydrocarbon rings. Non-limiting examples include
phenyl, naphthyl or tetrahydronaphthyl.
[0039] As used herein, the term "C.sub.1-6alkoxy" refers to
C.sub.1-6alkyl-O--, wherein C.sub.1-6alkyl is defined herein above.
Representative examples of C.sub.1-1alkoxy include, but are not
limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy,
tert-butoxy, pentyloxy, and hexyloxy.
[0040] As used herein, the term "Het.sup.2" or "heterocyclic ring"
refers to a saturated or unsaturated non-aromatic ring or ring
system, which is a 4-, 5-, 6-, or 7-membered monocyclic ring
comprising a) 1 or 2 heteroatoms selected from N, O and S; or b)
--C(O)-- and 1 or 2 heteroatoms selected from N and O. The
heterocyclic group can be attached via a heteroatom or a carbon
atom. Examples of heterocyclic rings include tetrahydrofuran (THF),
dihydrofuran, 1,4-dioxane, morpholine, 1,4-dithiane, piperazine,
piperidine, 1,3-dioxolane, imidazolidine, imidazoline, pyrroline,
pyrrolidine, tetrahydropyran, dihydropyran, oxathiolane,
dithiolane, 1,3-dioxane, 1,3-dithiane, oxathiane, pyrrolidinonyl,
oxazolidinonyl, and thiomorpholine.
[0041] The term "C.sub.3-7cycloalkyl" refers to a fully saturated
or unsaturated monocyclic hydrocarbon group of 3-7 carbon atoms.
Exemplary monocyclic hydrocarbon groups include, but are not
limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl,
cyclohexyl and cyclohexenyl.
[0042] As used herein, the term "Het.sup.1" or "heteroaryl ring"
refers to an aromatic ring system, which is a 5- or 6-membered
monocyclic ring comprising a) one oxygen or sulphur atom and
optionally one or two nitrogen atoms; or b) from one to four
nitrogen atoms. Typical Heteroaryl rings include 2- or 3-thienyl,
2- or 3-furyl, 2- or 3-pyrrolyl, 2-, 4-, or 5-imidazolyl, 3-, 4-,
or 5-pyrazolyl, 2-, 4-, or 5-thiazolyl, 3-, 4-, or 5-isothiazolyl,
2-, 4-, or 5-oxazolyl, 3-, 4-, or 5-isoxazolyl, 3- or
5-(1,2,4-triazolyl), and 4- or 5-(1,2,3-triazolyl), tetrazolyl,
pyrimidinyl and pyridinyl.
[0043] As used herein, the term "halogen" or "halo" refers to
fluoro, chloro, bromo, and iodo.
Embodiment 2
[0044] A compound of the formula (I) or a pharmaceutically
acceptable salt thereof:
##STR00003##
[0045] wherein R.sup.1 represents phenyl,
(C.sub.4-C.sub.7)cycloalkenyl or Het.sup.1, which R.sup.1 group may
be unsubstituted or substituted on one or two carbon atoms by
substituents R.sup.a, and may further be substituted on a nitrogen
atom with a substituent R.sup.a1;
[0046] each R.sup.a independently represents
(C.sub.1-C.sub.4)alkyl, halo, halo(C.sub.1-C.sub.4)alkyl, cyano,
hydroxy(C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy,
halo(C.sub.1-C.sub.4)alkoxy,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl, R.sup.6OC(O)--, or
R.sup.6OC(O)(C.sub.1-C.sub.4)alkyl-;
[0047] R.sup.a1 represents (C.sub.1-C.sub.4)alkyl,
hydroxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-,
aryl(C.sub.1-C.sub.4)alkyl or
R.sup.6OC(O)(C.sub.1-C.sub.4)alkyl-;
[0048] R.sup.2 represents (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.7)cycloalkyl, (C.sub.4-C.sub.7)cycloalkenyl, phenyl,
furanyl, thiazolyl or thienyl, which R.sup.2 may be unsubstituted
or substituted on from one to three carbon atoms with substituents
R.sup.b;
[0049] each R.sup.b independently represents
(C.sub.1-C.sub.4)alkyl, halo, halo(C.sub.1-C.sub.4)alkyl, cyano,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-,
(R.sup.6).sub.2NC(O)(C.sub.1-C.sub.4)alkyl- or
R.sup.6OC(O)(C.sub.1-C.sub.4)alkyl-;
[0050] X represents S, Z represents CHR.sup.4a and n represents 1;
or
[0051] X represents CHR.sup.4b, Z represents NR.sup.5 and n
represents 1; or
[0052] X represents CHR.sup.4b, Z represents CHR.sup.4a and n
represents 0 or 1;
[0053] each R.sup.3 independently represents methyl or ethyl;
[0054] when X represents S, R.sup.4 represents hydrogen,
(C.sub.1-C.sub.4)alkyl, halo(C.sub.1-C.sub.4)alkyl,
hydroxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl-,
amino(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-, or phenyl;
or
[0055] when X represents CHR.sup.4b, R.sup.4 represents hydrogen,
hydroxy, (C.sub.1-C.sub.4)alkyl, (C.sub.1-C.sub.4)alkoxy,
halo(C.sub.1-C.sub.4)alkyl, hydroxy(C.sub.1-C.sub.4)alkyl,
(C.sub.1-C.sub.4)alkoxy(C.sub.1-C.sub.4)alkyl-,
amino(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-, amino,
(C.sub.1-C.sub.4)alkylamino-, di[(C.sub.1-C.sub.4)alkyl]amino- or
phenyl;
[0056] R.sup.4a represents hydrogen, (C.sub.1-C.sub.4)alkyl,
halo(C.sub.1-C.sub.4)alkyl, amino(C.sub.1-C.sub.4)alkyl-,
(C.sub.1-C.sub.4)alkylamino(C.sub.1-C.sub.4)alkyl-,
di[(C.sub.1-C.sub.4)alkyl]amino(C.sub.1-C.sub.4)alkyl-,
Het.sup.2(C.sub.1-C.sub.4)alkyl-, or R.sup.6OC(O)--;
[0057] R.sup.4b represents hydrogen or methyl;
[0058] R.sup.5 represents hydrogen or (C.sub.1-C.sub.4)alkyl;
[0059] R.sup.6 represents hydrogen or (C.sub.1-C.sub.4)alkyl;
[0060] Het.sup.1 represents a 5- or 6-membered heteroaryl ring
comprising a) one oxygen or sulphur atom and optionally one or two
nitrogen atoms; or b) from one to four nitrogen atoms; and
[0061] Het.sup.2 represents a 4- to 7-membered heterocyclic ring
comprising 1 or 2 heteroatoms selected from N, O and S.
Embodiment 3
[0062] A compound according to Embodiment 1 or Embodiment 2,
wherein R.sup.1 represents phenyl, cyclohexenyl, thiazolyl,
pyrazolyl, thienyl, pyrimidin-2-yl or pyridine-2-yl and wherein
R.sup.1 may be unsubstituted or substituted on one or two carbon
atoms by substituents R.sup.a, and may further be substituted on a
nitrogen atom with a substituent R.sup.a1.
Embodiment 4
[0063] A compound according to any preceding Embodiment, wherein
R.sup.1 represents thiazol-2-yl, thiazol-4-yl, thien-2-yl or
pyrazol-4-yl, which R.sup.1 group may be unsubstituted or
substituted on one or two carbon atoms by substituents R.sup.a, and
may further be substituted on a nitrogen atom with a substituent
R.sup.a1
Embodiment 5
[0064] A compound according to any preceding Embodiment, wherein
R.sup.1 represents phenyl, which may be unsubstituted or
substituted by 1 or 2 substituents R.sup.a.
Embodiment 6
[0065] A compound according to Embodiment 5, wherein the R.sup.a
substitutents are in the 3-position, 2- and 3-positions, 3- and
4-positions, 3- and 5-positions or 3- and 6-positions.
Embodiment 7
[0066] A compound according to any preceding Embodiment, wherein
each R.sup.a independently represents (C.sub.1-C.sub.4)alkyl, halo,
halo(C.sub.1-C.sub.4)alkyl, hydroxy(C.sub.1-C.sub.4)alkyl, cyano,
(C.sub.1-C.sub.4)alkoxy, R.sup.6OC(O)--, or
R.sup.6OC(O)(C.sub.1-C.sub.4)alkyl-.
Embodiment 8
[0067] A compound according to any preceding Embodiment, wherein
R.sup.2 represents phenyl, furanyl, thiazolyl, or thienyl, which
R.sup.2 may be unsubstituted or substituted on from one to three
carbon atoms with substituents R.sup.b.
Embodiment 9
[0068] A compound according to any preceding Embodiment, wherein
each R.sup.b independently represents (C.sub.1-C.sub.2)alkyl, halo,
halo(C.sub.1-C.sub.2)alkyl, or cyano.
Embodiment 10
[0069] A compound according to any preceding Embodiment, wherein
each R.sup.b independently represents methyl, ethyl, bromo, chloro,
fluoro, trifluoromethyl, or cyano.
Embodiment 11
[0070] A compound according to any preceding Embodiment, wherein
each R.sup.3 represents methyl.
Embodiment 12
[0071] A compound according to any preceding Embodiment, wherein
R.sup.4 represents hydrogen, methyl, phenyl or HOCH.sub.2--.
Embodiment 13
[0072] A compound according to any preceding Embodiment, wherein Z
represents --CHR.sup.4a and R.sup.4a represents hydrogen or
methyl.
Embodiment 14
[0073] A compound according to any preceding Embodiment,
wherein:
[0074] X represents S, Z represents CHR.sup.4a and n represents 1;
or
[0075] X represents CHR.sup.4b, Z represents CHR.sup.4a and n
represents 1.
Embodiment 15
[0076] A compound according to any one of Embodiments 1 to 13,
wherein Z represents NR.sup.5 and R.sup.5 represents hydrogen.
Embodiment 16
[0077] A compound according to any preceding Embodiment, wherein
R.sup.6 represents hydrogen, methyl or ethyl.
Embodiment 17
[0078] A compound according to any preceding Embodiment, wherein
the compound is of formula (Ia):
##STR00004##
Embodiment 18
[0079] A compound according to any preceding Embodiment, wherein
the compound of formula (I) is racemic or has the stereochemistry
as shown in formula (Ia).
Embodiment 19
[0080] A compound of formula (I) or formula (Ia), which is selected
from the list consisting of:
Example 1.1
[0081]
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8-dihydro-1H-pyri-
mido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 1.4
[0081] [0082]
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[4-
',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 1.5
[0082] [0083]
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',-
5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 1.6
[0083] [0084]
10-(3-Chlorophenyl)-1,3-dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',5':3-
,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 1.7
[0084] [0085]
1,3-Dimethyl-5-phenyl-10-(m-tolyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrr-
olo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 1.8
[0085] [0086]
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(m-tolyl)-7,8-dihydro-1H-pyrimido[-
4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 1.9
[0086] [0087]
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-(m-tolyl)-7,8-dihydro-1H-pyrimid-
o[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 1.10
[0087] [0088]
1,3-Dimethyl-10-(5-methylthiophen-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[-
4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
Example 1.11
[0088] [0089]
1,3-Dimethyl-10-(4-methylthiophen-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[-
4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
Example 1.1.1
[0089] [0090]
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',-
5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 1.1.2
[0090] [0091]
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[4-
',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 1.1.3
[0091] [0092]
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[4',-
5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 1.1.4
[0092] [0093]
10-(3-Chlorophenyl)-1,3-dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',5':3-
,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 1.1.5
[0093] [0094]
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(m-tolyl)-7,8-dihydro-1H-pyrimido[-
4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 2.0
[0094] [0095]
7-((dimethylamino)methyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7-
,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-d-
ione;
Example 3.0
[0095] [0096]
10-(5-chlorofuran-2-yl)-1,3,8-trimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[-
4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 3.3
[0096] [0097]
1,3,8-trimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimid-
o[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 3.4
[0097] [0098]
3-(10-(5-Chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-hexahydro--
1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)benzonitrile;
Example 3.7
[0098] [0099]
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8-dihydro-
-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 3.8
[0099] [0100]
10-(5-Chlorofuran-2-yl)-5-(cyclohex-1-en-1-yl)-1,3-dimethyl-7,8-dihydro-1-
H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 3.9
[0100] [0101]
10-(5-Chlorofuran-2-yl)-5-(3,5-difluorophenyl)-1,3-dimethyl-7,8-dihydro-1-
H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 3.1.1
[0101] [0102]
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8-dihydro-
-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 3.1.2
[0102] [0103]
10-(5-chlorofuran-2-yl)-5-(cyclohex-1-en-1-yl)-1,3-dimethyl-7,8-dihydro-1-
H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 4.1
[0103] [0104]
10-(5-Chlorofuran-2-yl)-1,3-diethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',5-
':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 5
[0104] [0105]
3-Chloro-5-(10-(5-chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-h-
exahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)benzonitril-
e;
Example 6.1
[0105] [0106]
10-(2,3-Difluorophenyl)-1,3-dimethyl-5-phenyl-7,8,9,10-tetrahydropyrimido-
[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 6.2
[0106] [0107]
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimido-
[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 6.5
[0107] [0108]
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimi-
do[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 7.1
[0108] [0109]
5-(3-Fluorophenyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetrahyd-
ro pyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 7.4
[0109] [0110]
5-(3-fluorophenyl)-1,3,9-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetra-
hydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 7.5
[0110] [0111]
5-(3-fluorophenyl)-1,3,8-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetra-
hydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 7.6
[0111] [0112]
5-(3-fluorophenyl)-1,3,9-trimethyl-10-(4-methylthiazol-2-yl)-7,8,9,10-tet-
rahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 8.1
[0112] [0113]
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-m-tolyl-7,8,9,10-tetrahydropyrim-
ido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 8.3
[0113] [0114]
5-(3-Chlorophenyl)-10-(4-chlorothiazol-2-yl)-1,3-dimethyl-7,8,9,10-tetrah-
ydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 9.0
[0114] [0115]
3-(1,3-Dimethyl-10-(4-methylthiazol-2-yl)-2,4-dioxo-1,2,3,4,7,8,9,10-octa-
hydro pyrimido[4,5-a]indolizin-5-yl)benzonitrile;
Example 9.2
[0115] [0116]
3-(1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4,7,8,9,10-octahydropyrimido[4,5-
-a]indolizin-10-yl)benzonitrile; [0117]
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimi-
do[4',5':3,4]pyrrolo[1,2-b]pyridazine-2,4(1H,3H)-dione;
Example 10.2
[0117] [0118]
5-(3-Chlorophenyl)-1,3-dimethyl-10-(4-methylthiazol-2-yl)-7,8,9,10-tetrah-
ydropyrimido[4',5':3,4]pyrrolo[1,2-b]pyridazine-2,4(1H,3H)-dione;
Example 11
[0118] [0119]
1,3-dimethyl-9-(5-methylfuran-2-yl)-5-phenyl-8,9-dihydro-1H-pyrimido[4,5--
a]pyrrolizine-2,4(3H,7H)-dione;
Example 12.0
[0119] [0120]
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-phenyl-7,8-dihyd-
ro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 12.1
[0120] [0121]
10-(5-chlorofuran-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimethyl-
-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-
-dione;
Example 12.2
[0121] [0122]
3-(10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3,4-
,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)be-
nzonitrile;
Example 12.3
[0122] [0123]
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3-
,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)-
benzonitrile
Example 12.4
[0123] [0124]
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10-
H)-dione;
Example 13
[0124] [0125]
3-(10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-1,2,3,4,7,8,9,10-octa-
hydropyrimido[4,5-a]indolizin-5-yl)benzonitrile;
Example 13.1
[0125] [0126]
10-(4-Chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl--
7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 13.2
[0126] [0127]
1,3-Dimethyl-5,10-bis(4-methylthiazol-2-yl)-7,8,9,10-tetrahydropyrimido[4-
,5-a]indolizine-2,4(1H,3H)-dione;
Example 13.3
[0127] [0128]
10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8,9,10--
tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 13.4
[0128] [0129]
10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(2-methylthiazol-4-yl)-7,8,9,10--
tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 14
[0129] [0130]
10-(5-chlorofuran-2-yl)-8-((dimethylamino)methyl)-1,3-dimethyl-5-phenyl-7-
,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-d-
ione;
Example 15
[0130] [0131]
10-(5-Chlorofuran-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one;
Example 15.1
[0131] [0132]
10-(4-Chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl--
7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)--
dione;
Example 15.2
[0132] [0133]
10-(5-Chlorofuran-2-yl)-5-(3-ethoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-py-
rimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 15.3
[0133] [0134]
5-(3-Methoxyphenyl)-1,3-dimethyl-10-(4-methylthiazol-2-yl)-7,8-dihydro-1H-
-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 15.4
[0134] [0135]
10-(5-Chlorofuran-2-yl)-5-(3-methoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-p-
yrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 15.5
[0135] [0136]
10-(2-Chlorothiazol-4-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8-d-
ihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione-
;
Example 15.6
[0136] [0137]
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8-dih-
ydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 15.7
[0137] [0138]
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-imidazol-4-yl)-7,8-di-
hydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 15.8
[0138] [0139]
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-4-yl)-7,8-dih-
ydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 15.9
[0139] [0140]
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(5-methyl-1,3,4-oxadiazol-2-yl)-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one;
Example 15.10
[0140] [0141]
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(4-methyloxazol-2-yl)-7,8-dihydro--
1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 15.11
[0141] [0142]
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8-dih-
ydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 15.12
[0142] [0143]
10-(5-chlorofuran-2-yl)-5-(3-ethoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-py-
rimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 15.13
[0143] [0144]
10-(5-Chlorofuran-2-yl)-5-(3-methoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-p-
yrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 15.14
[0144] [0145]
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(5-methyl-1,3,4-oxadiazol-2-yl)-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one;
Example 15:15
[0145] [0146]
10-(5-chlorofuran-2-yl)-5-(4-(1-hydroxyethyl)thiazol-2-yl)-1,3-dimethyl-7-
,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-d-
ione;
Example 15:16
[0146] [0147]
10-(4-chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl--
7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)--
dione;
Example 16
[0147] [0148]
9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9-dihydro-1H-pyrimido[4,-
5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 16.1
[0148] [0149]
1,3-Dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyrimido[4,-
5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 17
[0149] [0150]
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(4-methylthiazol-
-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione;
Example 17.1
[0150] [0151]
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(1-methyl-1H-pyr-
azol-3-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2-
,4(3H,10H)-dione;
Example 17.2
[0151] [0152]
10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(2-methylthiazol-
-4-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione;
Example 18
[0152] [0153]
9,9-Difluoro-5-(3-fluorophenyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-7,8,-
9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 19
[0153] [0154]
8-(aminomethyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-
-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,-
10H)-dione;
Example 20
[0154] [0155]
3-(9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9-hexahydro--
1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile;
Example 20.1
[0155] [0156]
9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8,9-dihydr-
o-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 20.2
[0156] [0157]
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydro-1H-p-
yrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 21
[0157] [0158]
9-(4-Chlorothiazol-2-yl)-1,3,8,8-tetramethyl-5-phenyl-8,9-dihydro-1H-pyri-
mido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 21.1
[0158] [0159]
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3,8,8-tetramethyl-8,9-dihyd-
ro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 21.2
[0159] [0160]
9-(4-Chlorothiazol-2-yl)-1,3,8,8-tetramethyl-5-phenyl-8,9-dihydro-1H-pyri-
mido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 21.3
[0160] [0161]
3-(1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-2,4-dioxo-2,3,4,7,8,9-hex-
ahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile;
Example 21.4
[0161] [0162]
5-(3-Fluorophenyl)-1,3,8,8-tetramethyl-9-(4-methylthiazol-2-yl)-8,9-dihyd-
ro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 21.5
[0162] [0163]
1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyri-
mido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 21.6
[0163] [0164]
9-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3,8,8-tetramethyl-8,9-dihyd-
ro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 21.7
[0164] [0165]
5-(3-Fluorophenyl)-1,3,8,8-tetramethyl-9-(4-methylthiazol-2-yl)-8,9-dihyd-
ro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 21.8
[0165] [0166]
3-(1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-2,4-dioxo-2,3,4,7,8,9-hex-
ahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile;
Example 21.9
[0166] [0167]
1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyri-
mido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 22
[0167] [0168]
10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8,9-
,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 23
[0168] [0169]
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 23.1
[0169] [0170]
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-1,2-
,3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile;
Example 24
[0170] [0171]
7-((2H-1,2,3-triazol-2-yl)methyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-5--
phenyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 25
[0171] [0172]
2-(10-(5-Chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-hexahydro--
1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)thiazole-4-carboxyli-
c acid;
Example 26
[0172] [0173]
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8)methyl)methanesulfonamide;
Example 26.1
[0173] [0174]
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)-2-methoxyacetamide;
Example 26.2
[0174] [0175] Methyl
((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dioxo-
-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-8-
-yl)methyl) carbamate;
Example 26.3
[0175] [0176]
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)acetamide;
Example 26.4
[0176] [0177]
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)benzamide;
Example 27
[0177] [0178]
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-7-(hydroxymethyl)-1,3-dimethy-
l-8,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 27.1
[0178] [0179]
9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-5-phenyl-8,9-dihy-
dro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 27.2
[0179] [0180]
7-(hydroxymethyl)-1,3-dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihy-
dro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 27.3
[0180] [0181]
3-(9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3,-
4,7,8,9-hexahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile;
Example 28
[0181] [0182]
8-((1H-imidazol-1-yl)methyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-me-
thylthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thi-
azine-2,4(3H,10H)-dione;
Example 29
[0182] [0183]
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8-methylene-8,9--
dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione;
Example 30
[0183] [0184]
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxoox-
azolidin-3-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazine-2,4(3H,10H)-dione;
Example 30.1
[0184] [0185]
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxopy-
rrolidin-1-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazine-2,4(3H,10H)-dione;
Example 31
[0185] [0186]
3-(11-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9,10,11-oct-
ahydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepin-5-yl)benzonitrile;
Example 32
[0186] [0187]
11-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9,10,11-tetra-
hydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione;
Example 33
[0187] [0188]
1,3-Dimethyl-11-(4-methylthiazol-2-yl)-5-phenyl-8,9,10,11-tetrahydro-1H-p-
yrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione;
Example 34
[0188] [0189]
11-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9,10,11-tetrahydro-1H-p-
yrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione; and the
pharmaceutically acceptable salts thereof.
Embodiment 20
[0190] A compound of formula (I) or formula (Ia), which is selected
from the list consisting of:
Example 13a
[0191]
(R)-3-(10-(4-chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-1,2,3,4,7-
,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile;
Example 13.2a
[0191] [0192]
10-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8,9,10--
tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 13.3a
[0192] [0193]
10-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-(2-methylthiazol-4-yl)-7,8,9,10--
tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 13.1a
[0193] [0194]
10-(4-chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl--
7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 15
[0194] [0195]
10-(5-Chlorofuran-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one;
Example 15.11
[0195] [0196]
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8-dih-
ydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione;
Example 17
[0196] [0197]
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(4-methylthiazol-
-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione;
Example 17.1
[0197] [0198]
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(1-methyl-1H-pyr-
azol-3-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2-
,4(3H,10H)-dione;
Example 23a
[0198] [0199]
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 23.1a
[0199] [0200]
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-1,2-
,3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile;
Example 28a
[0200] [0201]
8-((1H-imidazol-1-yl)methyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-me-
thylthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thi-
azine-2,4(3H,10H)-dione; and the pharmaceutically acceptable salts
thereof.
Embodiment 21
[0202] A compound of formula (I) or formula (Ia), which is selected
from the list consisting of:
Example 13a
[0203]
(R)-3-(10-(4-chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-1,2,3,4,7-
,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile;
Example 13.1a
[0203] [0204]
10-(4-chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl--
7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 13.2a
[0204] [0205]
10-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8,9,10--
tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 13.3a
[0205] [0206]
10-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-(2-methylthiazol-4-yl)-7,8,9,10--
tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 15
[0206] [0207]
10-(5-Chlorofuran-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one;
Example 17
[0207] [0208]
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(4-methylthiazol-
-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione;
Example 23a
[0208] [0209]
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione;
Example 23.1a
[0209] [0210]
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-1,2-
,3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile;
and the pharmaceutically acceptable salts thereof.
[0211] As used herein, the term "an optical isomer" or "a
stereoisomer" refers to any of the various stereo isomeric
configurations which may exist for a given compound of the present
invention and includes geometric isomers. It is understood that a
substituent may be attached at a chiral center of a carbon atom.
The term "chiral" refers to molecules which have the property of
non-superimposability on their mirror image partner, while the term
"achiral" refers to molecules which are superimposable on their
mirror image partner. Therefore, the invention includes
enantiomers, diastereomers or racemates of the compound.
"Enantiomers" are a pair of stereoisomers that are
non-superimposable mirror images of each other. A 1:1 mixture of a
pair of enantiomers is a "racemic" mixture. The term is used to
designate a racemic mixture where appropriate. "Diastereoisomers"
are stereoisomers that have at least two asymmetric atoms, but
which are not mirror-images of each other. The absolute
stereochemistry is specified according to the Cahn-Ingold-Prelog
R-S system. When a compound is a pure enantiomer the
stereochemistry at each chiral carbon may be specified by either R
or S. Resolved compounds whose absolute configuration is unknown
can be designated (+) or (-) depending on the direction (dextro- or
levorotatory) which they rotate plane polarized light at the
wavelength of the sodium D line. Certain compounds described herein
contain one or more asymmetric centers or axes and may thus give
rise to enantiomers, diastereomers, and other stereoisomeric forms
that may be defined, in terms of absolute stereochemistry, as (R)-
or (S)-.
[0212] Depending on the choice of the starting materials and
procedures, the compounds can be present in the form of one of the
possible isomers or as mixtures thereof, for example as pure
optical isomers, or as isomer mixtures, such as racemates and
diastereoisomer mixtures, depending on the number of asymmetric
carbon atoms. The present invention is meant to include all such
possible isomers, including racemic mixtures, diasteriomeric
mixtures and optically pure forms. Optically active (R)- and
(S)-isomers may be prepared using chiral synthons or chiral
reagents, or resolved using conventional techniques. If the
compound contains a double bond, the substituent may be E or Z
configuration. If the compound contains a disubstituted cycloalkyl,
the cycloalkyl substituent may have a cis- or trans-configuration.
All tautomeric forms are also intended to be included.
[0213] As used herein, the terms "salt" or "salts" refers to an
acid addition or base addition salt of a compound of the invention.
"Salts" include in particular "pharmaceutical acceptable salts".
The term "pharmaceutically acceptable salts" refers to salts that
retain the biological effectiveness and properties of the compounds
of this invention and, which typically are not biologically or
otherwise undesirable. In many cases, the compounds of the present
invention are capable of forming acid and/or base salts by virtue
of the presence of amino and/or carboxyl groups or groups similar
thereto.
[0214] Pharmaceutically acceptable acid addition salts can be
formed with inorganic acids and organic acids, e.g., acetate,
aspartate, benzoate, besylate, bromide/hydrobromide,
bicarbonate/carbonate, bisulfate/sulfate, camphorsulfonate,
chloride/hydrochloride, chlortheophyllonate, citrate,
ethandisulfonate, fumarate, gluceptate, gluconate, glucuronate,
hippurate, hydroiodide/iodide, isethionate, lactate, lactobionate,
laurylsulfate, malate, maleate, malonate, mandelate, mesylate,
methylsulphate, naphthoate, napsylate, nicotinate, nitrate,
octadecanoate, oleate, oxalate, palmitate, pamoate,
phosphate/hydrogen phosphate/dihydrogen phosphate,
polygalacturonate, propionate, stearate, succinate,
sulfosalicylate, tartrate, tosylate and trifluoroacetate salts.
[0215] Inorganic acids from which salts can be derived include, for
example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric
acid, phosphoric acid, and the like.
[0216] Organic acids from which salts can be derived include, for
example, acetic acid, propionic acid, glycolic acid, oxalic acid,
maleic acid, malonic acid, succinic acid, fumaric acid, tartaric
acid, citric acid, benzoic acid, mandelic acid, methanesulfonic
acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic
acid, and the like. Pharmaceutically acceptable base addition salts
can be formed with inorganic and organic bases.
[0217] Inorganic bases from which salts can be derived include, for
example, ammonium salts and metals from columns I to XII of the
periodic table. In certain embodiments, the salts are derived from
sodium, potassium, ammonium, calcium, magnesium, iron, silver,
zinc, and copper; particularly suitable salts include ammonium,
potassium, sodium, calcium and magnesium salts.
[0218] Organic bases from which salts can be derived include, for
example, primary, secondary, and tertiary amines, substituted
amines including naturally occurring substituted amines, cyclic
amines, basic ion exchange resins, and the like. Certain organic
amines include isopropylamine, benzathine, cholinate,
diethanolamine, diethylamine, lysine, meglumine, piperazine and
tromethamine.
[0219] In another aspect, the present invention provides as
Embodiment 22, a compound of formula (I), or a pharmaceutically
acceptable salt thereof, according to any one of embodiments 1 to
21, in acetate, ascorbate, adipate, aspartate, benzoate, besylate,
bromide/hydrobromide, bicarbonate/carbonate, bisulfate/sulfate,
camphorsulfonate, caprate, chloride/hydrochloride,
chlortheophyllonate, citrate, ethandisulfonate, fumarate,
gluceptate, gluconate, glucuronate, glutamate, glutarate,
glycolate, hippurate, hydroiodide/iodide, isethionate, lactate,
lactobionate, laurylsulfate, malate, maleate, malonate, mandelate,
mesylate, methylsulphate, mucate, naphthoate, napsylate,
nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate,
pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate,
polygalacturonate, propionate, sebacate, stearate, succinate,
sulfosalicylate, sulfate, tartrate, tosylate trifenatate,
trifluoroacetate or xinafoate salt form.
Embodiment 23
[0220] A compound of formula (I), or a pharmaceutically acceptable
salt thereof, according to any one of Embodiments 1 to 21, which is
in sodium, potassium, ammonium, calcium, magnesium, iron, silver,
zinc, copper, isopropylamine, benzathine, cholinate,
diethanolamine, diethylamine, lysine, meglumine, piperazine or
tromethamine salt form.
[0221] The pharmaceutically acceptable salts of the present
invention can be synthesized from a basic or acidic moiety, by
conventional chemical methods. Generally, such salts can be
prepared by reacting free acid forms of these compounds with a
stoichiometric amount of the appropriate base (such as Na, Ca, Mg,
or K hydroxide, carbonate, bicarbonate or the like), or by reacting
free base forms of these compounds with a stoichiometric amount of
the appropriate acid. Such reactions are typically carried out in
water or in an organic solvent, or in a mixture of the two.
Generally, use of non-aqueous media like ether, ethyl acetate,
ethanol, isopropanol, or acetonitrile is desirable, where
practicable. Lists of additional suitable salts can be found, e.g.,
in "Remington's Pharmaceutical Sciences", 20th ed., Mack Publishing
Company, Easton, Pa., (1985); and in "Handbook of Pharmaceutical
Salts: Properties, Selection, and Use" by Stahl and Wermuth
(Wiley-VCH, Weinheim, Germany, 2002).
[0222] Any formula given herein is also intended to represent
unlabeled forms as well as isotopically labeled forms of the
compounds. Isotopically labeled compounds have structures depicted
by the formulas given herein except that one or more atoms are
replaced by an atom having a selected atomic mass or mass number.
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.11C, .sup.13C, .sup.14C, .sup.15N, .sup.18F .sup.31P,
.sup.32P, .sup.35S, .sup.36Cl, .sup.125I respectively. The
invention includes various isotopically labeled compounds as
defined herein, for example those into which radioactive isotopes,
such as .sup.3H and .sup.14C, or those into which non-radioactive
isotopes, such as .sup.2H and .sup.13C are present. Such
isotopically labelled compounds are useful in metabolic studies
(with .sup.14C), reaction kinetic studies (with, for example
.sup.2H or .sup.3H), detection or imaging techniques, such as
positron emission tomography (PET) or single-photon emission
computed tomography (SPECT) including drug or substrate tissue
distribution assays, or in radioactive treatment of patients. In
particular, an .sup.18F or labeled compound may be particularly
desirable for PET or SPECT studies. Isotopically-labeled compounds
of formula (I) can generally be prepared by conventional techniques
known to those skilled in the art or by processes analogous to
those described in the accompanying Examples and Preparations using
an appropriate isotopically-labeled reagents in place of the
non-labeled reagent previously employed.
[0223] Further, substitution with heavier isotopes, particularly
deuterium (i.e., .sup.2H or D) may afford certain therapeutic
advantages resulting from greater metabolic stability, for example
increased in vivo half-life or reduced dosage requirements or an
improvement in therapeutic index. It is understood that deuterium
in this context is regarded as a substituent of a compound of the
formula (I). The concentration of such a heavier isotope,
specifically deuterium, may be defined by the isotopic enrichment
factor. The term "isotopic enrichment factor" as used herein means
the ratio between the isotopic abundance and the natural abundance
of a specified isotope. If a substituent in a compound of this
invention is denoted deuterium, such compound has an isotopic
enrichment factor for each designated deuterium atom of at least
3500 (52.5% deuterium incorporation at each designated deuterium
atom), at least 4000 (60% deuterium incorporation), at least 4500
(67.5% deuterium incorporation), at least 5000 (75% deuterium
incorporation), at least 5500 (82.5% deuterium incorporation), at
least 6000 (90% deuterium incorporation), at least 6333.3 (95%
deuterium incorporation), at least 6466.7 (97% deuterium
incorporation), at least 6600 (99% deuterium incorporation), or at
least 6633.3 (99.5% deuterium incorporation).
[0224] Pharmaceutically acceptable solvates in accordance with the
invention include those wherein the solvent of crystallization may
be isotopically substituted, e.g. D.sub.2O, d.sub.6-acetone,
d.sub.6-DMSO.
[0225] Compounds of the invention, i.e. compounds of formula (I)
that contain groups capable of acting as donors and/or acceptors
for hydrogen bonds may be capable of forming co-crystals with
suitable co-crystal formers. These co-crystals may be prepared from
compounds of formula (I) by known co-crystal forming procedures.
Such procedures include grinding, heating, co-subliming,
co-melting, or contacting in solution compounds of formula (I) with
the co-crystal former under crystallization conditions and
isolating co-crystals thereby formed. Suitable co-crystal formers
include those described in WO 2004/078163. Hence the invention
further provides co-crystals comprising a compound of formula
(I).
[0226] As used herein, the term "pharmaceutically acceptable
carrier" includes any and all solvents, dispersion media, coatings,
surfactants, antioxidants, preservatives (e.g., antibacterial
agents, antifungal agents), isotonic agents, absorption delaying
agents, salts, preservatives, drug stabilizers, binders,
excipients, disintegration agents, lubricants, sweetening agents,
flavoring agents, dyes, and the like and combinations thereof, as
would be known to those skilled in the art (see, for example,
Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing
Company, 1990, pp. 1289-1329). Except insofar as any conventional
carrier is incompatible with the active ingredient, its use in the
therapeutic or pharmaceutical compositions is contemplated.
[0227] The term "a therapeutically effective amount" of a compound
of the present invention refers to an amount of the compound of the
present invention that will elicit the biological or medical
response of a subject, for example, reduction or inhibition of an
enzyme or a protein activity, or ameliorate symptoms, alleviate
conditions, slow or delay disease progression, or prevent a
disease, etc.
[0228] In one non-limiting embodiment, the term "a therapeutically
effective amount" refers to the amount of the compound of the
present invention that, when administered to a subject, is
effective to (1) at least partially alleviate, inhibit, prevent
and/or ameliorate a condition, or a disorder or a disease (i)
mediated by CFTR or (ii) associated with CFTR activity, or (iii)
characterized by activity (normal or abnormal) of CFTR or (2)
reduce or inhibit the activity of CFTR. In another non-limiting
embodiment, the term "a therapeutically effective amount" refers to
the amount of the compound of the present invention that, when
administered to a cell, or a tissue, or a non-cellular biological
material, or a medium, is effective to at least partially reducing
or inhibiting the activity of CFTR; or at least partially reducing
or inhibiting the expression of CFTR.
[0229] As used herein, the term "subject" refers to an animal.
Typically the animal is a mammal. A subject also refers to for
example, primates (e.g., humans, male or female), cows, sheep,
goats, horses, dogs, cats, rabbits, rats, mice, fish, birds and the
like. In certain embodiments, the subject is a primate. In yet
other embodiments, the subject is a human.
[0230] As used herein, the term "inhibit", "inhibition" or
"inhibiting" refers to the reduction or suppression of a given
condition, symptom, or disorder, or disease, or a significant
decrease in the baseline activity of a biological activity or
process.
[0231] As used herein, the term "treat", "treating" or "treatment"
of any disease or disorder refers in one embodiment, to
ameliorating the disease or disorder (i.e., slowing or arresting or
reducing the development of the disease or at least one of the
clinical symptoms thereof). In another embodiment "treat",
"treating" or "treatment" refers to alleviating or ameliorating at
least one physical parameter including those which may not be
discernible by the patient. In yet another embodiment, "treat",
"treating" or "treatment" refers to modulating the disease or
disorder, either physically, (e.g., stabilization of a discernible
symptom), physiologically, (e.g., stabilization of a physical
parameter), or both. In yet another embodiment, "treat", "treating"
or "treatment" refers to preventing or delaying the onset or
development or progression of the disease or disorder.
[0232] As used herein, a subject is "in need of" a treatment if
such subject would benefit biologically, medically or in quality of
life from such treatment.
[0233] As used herein, the term "a," "an," "the" and similar terms
used in the context of the present invention (especially in the
context of the claims) are to be construed to cover both the
singular and plural unless otherwise indicated herein or clearly
contradicted by the context.
[0234] All methods described herein can be performed in any
suitable order unless otherwise indicated herein or otherwise
clearly contradicted by context. The use of any and all examples,
or exemplary language (e.g. "such as") provided herein is intended
merely to better illuminate the invention and does not pose a
limitation on the scope of the invention otherwise claimed.
[0235] Any asymmetric atom (e.g., carbon or the like) of the
compound(s) of the present invention can be present in racemic or
enantiomerically enriched, for example the (R)-, (S)- or
(R,S)-configuration. In certain embodiments, each asymmetric atom
has at least 50% enantiomeric excess, at least 60% enantiomeric
excess, at least 70% enantiomeric excess, at least 80% enantiomeric
excess, at least 90% enantiomeric excess, at least 95% enantiomeric
excess, or at least 99% enantiomeric excess in the (R)- or
(S)-configuration. Substituents at atoms with unsaturated double
bonds may, if possible, be present in cis-(Z)- or
trans-(E)-form.
[0236] Accordingly, as used herein a compound of the present
invention can be in the form of one of the possible isomers,
rotamers, atropisomers, tautomers or mixtures thereof, for example,
as substantially pure geometric (cis or trans) isomers,
diastereomers, optical isomers (antipodes), racemates or mixtures
thereof.
[0237] Any resulting mixtures of isomers can be separated on the
basis of the physicochemical differences of the constituents, into
the pure or substantially pure geometric or optical isomers,
diastereomers, racemates, for example, by chromatography and/or
fractional crystallization.
[0238] Any resulting racemates of final products or intermediates
can be resolved into the optical antipodes by known methods, e.g.,
by separation of the diastereomeric salts thereof, obtained with an
optically active acid or base, and liberating the optically active
acidic or basic compound. In particular, a basic moiety may thus be
employed to resolve the compounds of the present invention into
their optical antipodes, e.g., by fractional crystallization of a
salt formed with an optically active acid, e.g., tartaric acid,
dibenzoyl tartaric acid, diacetyl tartaric acid, di-O,O'-p-toluoyl
tartaric acid, mandelic acid, malic acid or camphor-10-sulfonic
acid. Racemic products can also be resolved by chiral
chromatography, e.g., high pressure liquid chromatography (HPLC)
using a chiral adsorbent.
[0239] Furthermore, the compounds of the present invention,
including their salts, can also be obtained in the form of their
hydrates, or include other solvents used for their crystallization.
The compounds of the present invention may inherently or by design
form solvates with pharmaceutically acceptable solvents (including
water); therefore, it is intended that the invention embrace both
solvated and unsolvated forms. The term "solvate" refers to a
molecular complex of a compound of the present invention (including
pharmaceutically acceptable salts thereof) with one or more solvent
molecules. Such solvent molecules are those commonly used in the
pharmaceutical art, which are known to be innocuous to the
recipient, e.g., water, ethanol, and the like. The term "hydrate"
refers to the complex where the solvent molecule is water.
[0240] The compounds of the present invention, including salts,
hydrates and solvates thereof, may inherently or by design form
polymorphs.
[0241] Typically, the compounds of formula (I) can be prepared
according to the Schemes provided infra.
[0242] Compounds of formula (I), wherein X is S, Z is CH.sub.2, n
is 1, R.sup.4 is H, and R.sup.1, R.sup.2 and R.sup.3 are as defined
hereinabove, may be prepared according to Scheme 1.
##STR00005##
[0243] Step 1(i) An intermediate of formula (III) may be prepared
from an intermediate of formula (II) by removal of the trityl
moiety in the presence of a suitable acid, such as HCl, or acid and
reducing agent, such as trifluoroacetic acid and
triethylsilane.
[0244] Step 1(ii) A compound of formula (I) may be prepared from an
intermediate of formula (III) by reaction with a suitable aldehyde
in the presence of a Lewis acid, such as bismuth triflate.
[0245] Intermediates of formula (II) may be prepared according to
Scheme 2.
##STR00006##
[0246] wherein R represents
##STR00007##
R.sup.a represents methyl or i-propyl; E represents I and G.sup.1
represents boronic acid or boronate ester; or E represents
B(OH).sub.2 and G.sup.1 represents Cl, Br, I or OTf
[0247] Step 2(i) An intermediate of formula (IV) may be prepared by
reaction of a suitable pyrimidine-2,4(1H,3H)-dione with
1-(isocyanomethyl)sulfonyl)-4-methylbenzene in the presence of a
suitable base, such as sodium hydride in DMSO/THF or potassium
t-butoxide in 5-methyl-tetrahydrofuran.
[0248] Step 2(ii) An intermediate of formula (V) may be prepared by
N-alkylation of an intermediate of formula (IV) with a compound
R--Cl.
[0249] Step 2(iii) An intermediate of formula (VI) maybe prepared
from an intermediate of formula (V) by reaction with a suitable
electrophile, such as I.sub.2 or B(OR.sup.a).sub.3, in the presence
of a suitable base, such as LDA or BuLi. The skilled person will
recognize that the C5-lithiated species may either be pre-formed or
formed in-situ in the presence of the appropriate electrophile.
[0250] Step 2(iv) An intermediate of formula (II) may be prepared
from an intermediate of formula (VI) via a Suzuki cross-coupling
reaction with a compound of formula R.sup.1-G.sup.1, in the
presence of a suitable palladium catalyst, such as
tetrakis-(triphenylphosphine)palladium (0) or
1,1''bis(di-t-butylphosphino)ferrocene palladium dichloride
(Johnson Matthey PD-118).
[0251] Compounds of formula (I) wherein Z is CR.sup.4a, n is 1, and
X, R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are as defined
hereinabove, may be prepared according to Scheme 3.
##STR00008##
[0252] wherein LG represents a suitable leaving group such as
mesylate or triflate; G.sup.2 represents halide, preferably
bromide; and Y represents trityl or acetyl.
[0253] Step 3(i) An intermediate of formula (VIII) may be prepared
by reaction of an intermediate of formula (VII) with the required
amino alcohol in the presence of a suitable base, such as
triethylamine in ethanol.
[0254] Step 3(ii) An intermediate of formula (IX), may be prepared
from an intermediate of formula (VIII) by reaction with a suitable
agent, such as mesyl chloride, mesyl anhydride or triflic
anhydride, optionally in the presence of a suitable catalyst such
as DMAP. Alternatively, where R.sup.4 in the compound of formula
(I) is --CH.sub.2OH, an intermediate of formula (IX) may be
prepared from an intermediate of formula (VII) according to the
process described in Steps 3(i), 3(vi) and 3(ii), using the
required amino diol and a suitable protecting group for the primary
hydroxy group, such as TBDMS, followed by reaction of the secondary
alcohol as described in Step 3(ii).
[0255] Step 3(iii) Direct displacement of the activated secondary
alcohol (IX) to provide an intermediate of formula (X) can be
achieved by reaction with a suitable sulphur based nucleophile,
such as potassium thioacetate or the sodium salt of triphenylmethyl
mercaptan.
[0256] Step 3(iv) When Y is acetyl, deprotection of intermediate
(X), can be achieved using NaBH.sub.4 in ethanol. When Y is trityl,
intermediate (X) may be deprotected using the conditions described
in Step 1(i).
[0257] Step 3(v) Cyclisation to provide a compound of formula (I)
can be achieved by a process analogous to that described in Step
1(ii).
[0258] Intermediates of formula (VII) may be prepared according to
Scheme 4.
##STR00009##
[0259] Steps 4(i) and (ii) Intermediates of formula (XV) may be
prepared in a two-step process by reaction of acetic anhydride and
dimethyl urea, followed by a Friedel-Crafts acylation using the
required acyl chloride in the presence of a suitable Lewis acid,
such as zinc chloride or aluminium trichloride.
[0260] Step 4(iii) An intermediate of formula (VII) may be prepared
by halogenation of an intermediate of formula (XV) in the presence
or absence of a radical initiator. Suitably, bromine in chloroform
may be used.
[0261] Compounds of formula (I), wherein X is S, Z is CR.sup.4a,
R.sup.4a is amino-CH.sub.2--,
(C.sub.1-C.sub.4)alkylamino-CH.sub.2-- or
di[(C.sub.1-C.sub.4)alkyl]amino-CH.sub.2--, n is 1, R.sup.4 is H,
and R.sup.1, R.sup.2 and R.sup.3 are as defined hereinabove, may be
prepared according to Scheme 5.
##STR00010## ##STR00011##
[0262] wherein each R is independently selected from H and
(C.sub.1-C.sub.4)alkyl; and LG represents a suitable leaving group
such as mesylate or triflate.
[0263] Step 5(i) and (ii) An intermediate of formula (XVII) may be
prepared from an intermediate of formula (VII) via processes
analogous to those described in step 3(i) followed by step
3(ii).
[0264] Step 5(iii) An intermediate of formula (XVII) may be
converted to the corresponding secondary amine of formula (XVIII)
by displacement of the leaving group with a suitable nucleophile,
R.sub.2NH.
[0265] Steps 5(iv) and (v) A compound of formula (I) may be
prepared from an intermediate of formula (XVIII) via processes
analogous to those described in Step 1(i) followed by Step
1(ii).
[0266] Compounds of formula (I), wherein X is CH.sub.2, Z is NH, n
is 1, R.sup.4 is H and R.sup.1, R.sup.2 and R.sup.3 are as defined
hereinabove, may be prepared according to Scheme 6.
##STR00012## ##STR00013##
[0267] wherein ZNHNH.sub.2 is benzyl carbazate.
[0268] Step 6(i) An intermediate of formula (XX) may be prepared by
reaction of an intermediate of formula (VII) with benzyl carbazate
using conditions analogous to those described in step 3(i)
[0269] Step 6(ii) An intermediate of formula (XXI) may be prepared
by N-alkylation of an intermediate of formula (XX) with
3-bromopropan-1-ol in the presence of a suitable base, such as
potassium carbonate.
[0270] Step 6(iii) Oxidation of the alcohol intermediate of formula
(XXI) to provide the corresponding acid intermediate of formula
(XXII) can suitably be achieved using TPAP/NMO conditions
[0271] Step 6(iv) An intermediate of formula (XXIII) may be
prepared by cyclization of (XXII), suitably using polyphosphoric
acid (PPA).
[0272] Step 6(v) An enol-triflate of formula (XXIV) may be prepared
by reaction of an intermediate of formula (XXIII) with triflic
anhydride in the presence of a base, such as lutidine.
[0273] Step 6(vi) An intermediate of (XXV) may be prepared by
reaction of an enol-triflate of formula (XXIV) with a suitable
stannane, such as R.sup.2-tributyl tin in the presence of a
suitable palladium catalyst such as
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) and
suitable additives such as LiCl and CuI.
[0274] Step 6(vii) A compound of formula (I) may be prepared from
an intermediate of formula (XXV) by transfer hydrogenation, using
ammonium formate in the presence of palladium on carbon in a
suitable solvent such as ethanol.
[0275] Compounds of formula (I), wherein X and Z are CH.sub.2, n is
1, R.sup.4 is H, and R.sup.1, R.sup.2 and R.sup.3 are as defined
hereinabove, may be prepared according to Scheme 7. When R.sup.2 is
2-furyl, a compound of formula (I) may be prepared according to
steps 7(iv) and 7(v)
##STR00014##
[0276] Steps 7(i), (ii) and (iii) may be carried out in an
analogous fashion to the reactions described in Steps 6(v), (vi)
and (vii). Step 7(iii) may alternatively be carried out using
hydrogenation under palladium or platinum catalysis conditions,
such as palladium on carbon or platinum on carbon in ethanol.
[0277] Step 7(iv) An intermediate of formula (XXIX) may be prepared
by reduction of a compound of formula (XXVI) using a suitable
reducing agent such as NaBH.sub.4 in a suitable solvent such as
methanol.
[0278] Step 7(v) When R.sup.2 is a 2-furyl moiety, a compound of
formula (I) may be prepared directly from an intermediate of
formula (XXIX) by gold(III) catalysis.
[0279] Step 7(vi) An intermediate of formula (XXX) may be prepared
by reaction of an intermediate of formula (XXVII) with a suitable
boron species, such as bis(pinacolato)diboron, in the presence of a
suitable palladium catalyst, such as PdCl.sub.2(PPh.sub.3).sub.2,
and KOPh.
[0280] Step 7(vii) An intermediate of formula (XXVIII) may
alternatively be prepared from an intermediate of formula (XXX) by
a Suzuki cross-coupling reaction in an analogous fashion to the
reaction described in Step 2(iv).
[0281] An intermediate of formula (XXVI) may be prepared according
to Scheme 8.
##STR00015##
[0282] Step 8(i) An intermediate of formula (XXXI) may be prepared
by fluoride- or acid-catalysed deprotection of an intermediate of
formula (II), such as TBAF in THF.
[0283] Step 8(ii) N-alkylation of an intermediate of formula (XXXI)
to give an intermediate of formula (XXXII) may be carried out under
conditions analogous to those described in Step 2(ii), such as
caesium carbonate in DMF.
[0284] Step 8(iii) Ester hydrolysis of an intermediate of formula
(XXXII) to give an intermediate of formula (XXXIII) may be carried
out using a suitable base, such as LiOH in THF/H.sub.2O.
[0285] Step 8(iv) An intermediate of formula (XXVI) may be prepared
by cyclisation of an intermediate of formula (XXXIII) under
conditions analogous to those described in Step 6(iv), or by using
propane phosphonic acid anhydride (T3P.RTM.) in ethyl acetate or
DMF.
[0286] Step 8(v) Alternatively, an intermediate of formula (XXXII)
may be prepared from an intermediate of formula (VII) using the
required amino ester in the presence of a suitable base, such as
triethylamine, in a suitable solvent such as ethanol.
[0287] Compounds of formula (I), wherein X and Z are CH.sub.2, n is
0, and R.sup.1, R.sup.2 and R.sup.3 are as defined hereinabove, may
also be prepared according to Schemes 7 and 8, using the
corresponding .omega.-amino ester.
[0288] Alternatively, an intermediate of formula (XXVIII), may be
prepared according to Scheme 9.
##STR00016##
[0289] Step 9(i) and (ii) An intermediate of formula (XXXIV) may be
prepared by reaction of an intermediate of formula (VII) with an
amino acetal followed by acid catalyzed cyclisation, suitably using
HCl.
[0290] Step 9(iii) An intermediate of formula (XXVIII) may be
prepared from an intermediate of formula (XXXIV) using a Heck
reaction under palladium catalysis, suitably using palladium
acetate in the presence of tri-tert-butylphosphine
tetrafluorohydroborate in dimethylacetamide (DMA).
[0291] Intermediates of formula (XIV) may also be prepared
according to scheme 10.
##STR00017##
[0292] wherein LG is a leaving group such as mesylate or triflate,
preferably triflate.
[0293] Step 10(i) An intermediate of formula (XXXV) may be prepared
by treatment of the appropriate commercially available alcohol by
treatment with an appropriate sulfonylating agent in the presence
of a suitable base, such as trifluoromethanesulfonic anhydride in
the presence of 2,6-lutidine.
[0294] Step 10(ii) An intermediate of formula (XXXVI) may be
prepared from an intermediate of formula (XXXI) by treatment with
intermediate (XXXV) and a suitable base, such as sodium hydride in
DMF.
[0295] Step 10(iii) An intermediate of formula (XIV) may be
prepared from an intermediate of formula (XXXVI) by treatment with
an appropriate acid, such as HCl in diethyl ether.
[0296] Further compounds of formula (I) wherein Z is CR.sup.4a, n
is 1, and X, R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are as defined
hereinabove, may be prepared by derivatisation of another compound
of formula (I) where R.sup.4 is CH.sub.2OH according to Scheme
11.
##STR00018##
[0297] wherein LG represents a suitable leaving group such as
mesylate or triflate.
[0298] Step 11(i) An intermediate of formula (XXXVII) may be
prepared by reaction of a compound of formula (I) in the manner
described in Step 3(ii).
[0299] Step 11 (ii) A compound of formula (I) may be prepared by
reaction of an intermediate of formula (XXXVII) by treatment with a
nucleophile and application of heat in a suitable solvent, such as
dimethylamine in THF.
[0300] Step 11 (iii) Compounds of formula (I) may also be prepared
directly by direct treatment of another compound of formula (I)
with an appropriate nucleophile in the presence of a sulfonic
anhydride in a suitable solvent, such as sodium imidazolide in the
presence of triflic anhydride in DCM.
[0301] Compounds of formula (I), wherein X is CF.sub.2 and Z is
CH.sub.2, n is 1, R.sup.4 is H, and R.sup.1, R.sup.2 and R.sup.3
are as defined hereinabove, may be prepared according to Scheme
12.
##STR00019##
[0302] Step 12(i) An intermediate of formula (XXXVIII) may be
prepared from an intermediate of formula (XXVI) by treatment with
an appropriate fluorinating agent and an appropriate base in the
presence of an appropriate Lewis acid, such as
N-fluorobenzenesulfonimide and potassium hexamethyldisilazane in
the presence of manganese (II) bromide.
[0303] Steps 12(ii) and 12(iii) may be carried out by analogous
reactions to those described in Steps 7(iv) and 7(v).
[0304] Compounds of formula (I), wherein X and Z are CH.sub.2, n is
0, 1 or 2 and R.sup.1, R.sup.2 and R.sup.3 are as defined
hereinabove, may be prepared according to Scheme 13.
##STR00020##
[0305] wherein G.sub.2 represents halide, preferably bromide or
iodide, and Alk represents methyl or ethyl.
[0306] Step 13(i) For n=1 or 2, N-alkylation of an intermediate of
formula (XXXI) with a commercially available halide to give an
intermediate of formula (XLI) may be carried out under conditions
analogous to those described in Step 2(ii), such as cesium
carbonate in DMF. Alternatively, for n=0, N-alkylation of an
intermediate of formula (XXXI) to give an intermediate of formula
(XLI) may be carried out by treatment by treatment with a suitable
acrylate ester and a suitable base, such as methyl acrylate and
DBU.
[0307] Step 13(ii) Alternatively, an intermediate of formula (XLI)
may be prepared from an intermediate of formula (VII) using the
required commercially available amino ester in the presence of a
suitable base in a suitable solvent, such as triethylamine in
ethanol.
[0308] Step 13(iii) Ester hydrolysis of an intermediate of formula
(XLI) may be carried out to give an intermediate of formula (XLII)
by use of an appropriate base, such as lithium hydroxide.
[0309] Step 13(iv) An intermediate of formula (XLIII) may be
prepared from an intermediate of formula (XLII) by treatment with
N,O-dimethylhydroxylamine and an appropriate coupling reagent in
the presence of a suitable base, such as T3P in EtOAc or DMF in the
presence of DIPEA.
[0310] Step 13(v) Alternatively, an intermediate of formula (XLIII)
may be directly prepared from an intermediate of formula (XLI) by
treatment with N,O-dimethylhydroxylamine and a suitable base, such
as isopropylmagnesium bromide in THF.
[0311] Step 13(vi) An intermediate of formula (XLIV) may be
prepared from an intermediate of formula (XLIII) by reaction with
an appropriate halo compound R.sub.2-G.sub.2 in the presence of an
appropriate metallating reagent, such as isopropylmagnesium
chloride-lithium chloride complex.
[0312] Step 13(vii) An intermediate of formula (XLV) may be
prepared from an intermediate of formula (XLIV) by treatment with
an appropriate reducing agent, such as sodium borohydride-lithium
chloride in methanol.
[0313] Step 13(viii) A compound of formula (I) may be prepared from
an intermediate of formula (XLV) by treatment with an appropriate
activating agent and an appropriate base, such as triflic anhydride
in the presence of triethylamine.
[0314] Alternatively, compounds of formula (I) may be prepared from
and intermediate of formula (XLIV) by Steps 13(ix) and 13(x).
[0315] Step 13(ix) An intermediate of formula (XXVIII) may be
prepared by treatment of an intermediate of formula (XLIV) by
treatment with an appropriate acid, such as triflic acid.
[0316] Step 13(x) may be carried out using hydrogenation under
palladium or platinum catalysis conditions, such as palladium on
carbon or platinum on carbon in ethanol, analogously to Step
7(ii).
[0317] Compounds of formula (I), wherein Z is CHR.sup.4s, n is 0,
and R.sup.1, R.sup.2 and R.sup.3 are as defined hereinabove, may be
prepared according to Scheme 14.
##STR00021##
[0318] wherein G.sub.3 represents an appropriate metal or metalloid
residue, such as trialkylstannyl, PG represents an appropriate
protecting group, such as trialkylsilyl, and LG represents a
leaving group, such as mesylate.
[0319] Where X.dbd.CH.sub.2 and Z.dbd.CHCH.sub.2OH, compounds of
formula (I) may be prepared according to Steps 14(i)-14(vi). Where
X.dbd.CMe.sub.2 and Z.dbd.CH.sub.2, compounds of formula (I) may be
prepared according to Steps 14(vii)-14(x).
[0320] Step 14(i) An intermediate of formula (XLVII) may be
prepared from an intermediate of formula (XLVI) by treatment with
an appropriate allylating agentin the presence of an appropriate
lewis acid, such as tri-n-butylallylstannane in the presence of
boron trifluoride in THF.
[0321] Step 14(ii) An intermediate of formula (XLVIII) may be
prepared from an intermediate of formula (XLVII) by treatment with
an appropriate dihydroxylating reagent in an appropriate solvent,
such as osmium tetroxide and N-methylmorpholine oxide in
acetonitrile/water.
[0322] Step 14(iii) An intermediate of formula (XLIX) may be
prepared from an intermediate of formula (XLVIII) by O-protection
with a compound PG-Cl in the presence of a suitable base, such as
tertbutyldimethylsillylchloride in the presence of imidazole.
[0323] Step 14(iv) An intermediate of formula (L) may be prepared
from an intermediate of formula (XLIX) by an analogous reaction to
Step 3(ii).
[0324] Step 14(v) An intermediate of formula (LI) may be prepared
from an intermediate of formula (L) by treatment with a suitable
base in a suitable solvent, such as sodium hydride in THF.
[0325] Step 14(vi) A compound of formula (I) may be prepared from
an intermediate of formula (LI) by removal of the protecting group
in the presence of a suitable acid, such as TFA in methanol/water
mixtures.
[0326] Step 14(vii) An intermediate of formula (LII) may be
prepared from an intermediate of formula (XLVI) by treatment with
the commercially available silyl ketene acetal in the presence of a
suitable lewis acid, such as boron trifluoride.
[0327] Step 14(viii) An intermediate of formula (LIII) may be
prepared from an intermediate of formula (LII) by treatment with an
appropriate reducing agent in an appropriate solvent, such as
sodium borohydride-lithium chloride in methanol.
[0328] Step 14(ix) An intermediate of formula (LIV) may be prepared
from an intermediate of formula (LIII) by an analogous reaction to
Step 3(ii)
[0329] Step 14(x) A compound of formula (I) may be prepared from an
intermediate of formula (LIV) by an analogous reaction to Step
14(v).
[0330] Intermediates of formula (XLVI) may be prepared according to
Scheme 15.
##STR00022##
[0331] wherein G.sub.2 represents halide, preferably bromide or
iodide, and M represents a metal residue, preferably magnesium
halide.
[0332] Step 15(i) An intermediate of formula (LV) may be prepared
from an intermediate of formula (XXXI) by reaction with a suitable
formylating agent, such as POCOl.sub.3 in DMF
[0333] Step 15(ii) An intermediate of formula (XLVI) may be
prepared from an intermediate of formula (LV) by treatment with an
organometallic compound R.sup.2-M, which itself may by prepared
directly prior to use by treatment of a compound R.sup.2-G.sub.2
with a metallating agent, such as isopropylmagnesium
chloride-lithium chloride.
[0334] Compounds of formula (I), wherein X is C.dbd.CH.sub.2, Z is
CHR.sup.4s, n is 0, and R.sup.1, R.sup.2, R.sup.3 and R.sup.4a are
as defined hereinabove, may be prepared according to Scheme 16.
##STR00023##
[0335] wherein LG is a leaving group such as mesylate.
[0336] Step 16(i) An intermediate of formula (LVI) may be prepared
from an intermediate of formula (XLVI) by treatment with dimethyl
malonate in the presence of an appropriate base and an appropriate
Lewis acid, such as dimethyl malonate in the presence of sodium
hexamethyldisilazane and boron trifluoride.
[0337] Steps 16(ii), 1(iii) and 16(iv) may be carried out in by
reactions analogous to Steps 15(viii), 3(ii) and 14(v)
respectively.
[0338] Compounds of formula (I), wherein X is S and Z is H.sub.2, n
is 1, R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.a and Het.sup.2 are
as defined hereinabove and W is C.dbd.O or SO.sub.2, may be
prepared according to Scheme 17.
##STR00024##
[0339] wherein G.sub.2 is halide, preferably bromide
[0340] Step 17(i) An intermediate of formula (LX) may be prepared
by treatment of an intermediate of formula (XXXI) with an
intermediate of formula (LIX) and a suitable base in a suitable
solvent, such as cesium carbonate in DMF.
[0341] Step 17(ii) An intermediate of formula (LXI) may be prepared
from an intermediate of formula (LX) by reactions analogous to
Steps 1(i) and 1(ii).
[0342] Step 17(iii) Deprotection of the Phth protecting group may
be carried out by using ethanolamine and the application of heat to
give an intermediate of formula (LXII).
[0343] Step 17(iv) Compounds of formula (N) may be prepared by
treatment of an intermediate of formula (LXII) with a suitable
acylating agent or sulfonylating agent in the presence of a
suitable base in a suitable solvent, such as acetyl chloride or
methanesulfonyl chloride in the presence of DIPEA in DCM.
[0344] Step 17(v) Further compounds of formula (I) may be prepared
from an appropriate compound of formula (I) where R.sup.a is
halo(C1-C4)alkyl or halo(C1-C4)alkoxy by treatment with an
appropriate base in an appropriate solvent, such as cesiumsodium
hydride in THF.
[0345] An intermediate of formula (LIX) may be prepared according
to Scheme 18.
##STR00025##
[0346] wherein G.sub.2 is halide, preferably bromide
[0347] Step 18(i) An intermediate of formula (LXIII) may be
prepared from the commercially available epoxide by treatment with
thiourea in an appropriate solvent, such as methanol.
[0348] Step 18(ii) An intermediate of formula (LIX) may be prepared
from an intermediate of formula (LXIII) by treatment with an
appropriate trityl halide in an appropriate solvent, such a trityl
bromide in DCM.
[0349] Compounds of formula (I), wherein X is CHR.sup.4b and Z is
CHR.sup.4a, n is 1, R.sup.1, R.sup.2 and R.sup.3 are as defined
hereinabove, may be prepared according to Scheme 19.
##STR00026##
[0350] wherein LG is a leaving group such as mesylate or triflate,
preferably triflate, G.sub.2 is a halide, preferably bromide or
iodide and PG is a protecting group such as trialkylsilyl.
[0351] Step 19(i) An intermediate of formula (LXV) may be prepared
from an intermediate of formula (XXXI) and an intermediate of
formula (LXIV) by treatment with a suitable base in a suitable
solvent, such as potassium tert-butoxide in THF.
[0352] Step 19(ii) An intermediate of formula (LXVI) may be
prepared from an intermediate of formula (LXV) by base-mediated
hydrolysis with a suitable base, such as sodium hydroxide.
[0353] Step 19(iii), Step 19(iv) and 19(v) may be carried out by
analogous reactions to Steps 14(iii), 13(iv) and 13(vi)
respectively.
[0354] Step 19(vi) An intermediate of formula (LXX) may be prepared
from an intermediate of formula (LXIX) by treatment with a suitable
acid in a suitable solvent, such as pyridinium hydrochloride in
methanol and water.
[0355] Step 19(vii) may be carried out by an analogous reaction to
Step 7(iii).
[0356] An intermediate of formula (LXIV) may be prepared according
to Scheme 20.
##STR00027##
[0357] wherein LG is a leaving group such as mesylate or triflate,
preferably triflate.
[0358] Step 20(i) An intermediate of formula (LXXI) may be prepared
from the commerically available dihydroxyacetone by treatment with
(carbethoxymethylene)triphenylphosphorane in a suitable solvent,
such as DCM
[0359] Step 20(ii) An intermediate of formula (LXXII) may be
prepared from an intermediate of formula (LXXI) by hydrogenation
over a suitable catalyst, such as palladium on charcoal in
2-methyltetrahydrofuran
[0360] Step 20(iii) An intermediate of formula (LXIV) may be
prepared from an intermediate of formula (LXXII) by treatment with
a suitable activating agent in the presence of a suitable base,
such as trifluoromethanesulfonic anhydride in the presence of
2,6-lutidine
[0361] Compounds of formula (I), wherein X is CH.sub.2 and Z is
CHR.sup.4a, n is 1, R.sup.1, R.sup.2, R.sup.3 and Het.sup.2 are as
defined hereinabove, may be prepared according to Scheme 21.
##STR00028## ##STR00029##
[0362] wherein LG is a leaving group such as mesylate.
[0363] Steps 21(v) and 21(vi) may be carried out in a similar
manner to Steps 7(iv) and 7(v).
[0364] Step 21(i) An intermediate of formula (LXXIII) may be
prepared from an intermediate of formula (VII) in a similar manner
to Step 3(i) using the appropriate commercially available
aminodiester.
[0365] Step 21(ii) An intermediate of formula (LXXIV) may be
prepared from an intermediate of formula (LXXIII) by hydrogenolysis
under palladium catalysis conditions, such as palladium on carbon
in ethanol.
[0366] Step 21(iii) An intermediate of formula (LXXV) may be
prepared from an intermediate of formula (LXXIV) by an analogous
reaction to Step 8(iv).
[0367] Step 21(iv) An intermediate of formula (LXXVI) may be
prepared from an intermediate of formula (LXXV) by treatment with
an appropriate alkylating agent in the presence of a suitable base
with the application of heat in a suitable solvent, such as
dimethyl sulfate in the presence of potassium carbonate in wet
acetone.
[0368] Steps 21(v) and 21(vi) may be carried out by analogous
reactions to those described in Steps 7(iv) and 7(v).
[0369] Step 21(vii) An intermediate of formula (LXXIX) may be
prepared from an intermediate of formula (LXXVIII) by treatment
with a suitable reducing agent, such as lithium aluminium hydride
in THF.
[0370] Step 21(viii) An intermediate of formula (LXXX) may be
prepared from an intermediate of formula (LXXIX) by treatment with
an appropriate activating agent in the presence of a suitable base,
such as methansulfonyl chloride in the presence of
triethylamine.
[0371] Step 21(ix) A compound of formula (I) may be prepared from
an intermediate of formula (LXXX) by treatment with an appropriate
heterocycle Het.sup.2-H in the presence of a suitable base, such as
potassium carbonate in DMF.
[0372] Compounds of formula (I), wherein X is CH.sub.2 or CHMe, Z
is CH.sub.2, n is 1, R.sup.4 is H or Me, and R.sup.1, R.sup.2 and
R.sup.3 are as defined hereinabove, may be prepared according to
Scheme 22. When R.sup.2 is 2-furyl, compounds of formula (I) may be
prepared according to steps 7(iv) and 7(v)
##STR00030##
[0373] Steps 22(i) to 22(vii) may be carried out by analogous
reactions to those detailed in Scheme 7.
[0374] Intermediates of formula (LXXXI) may be prepared according
to Scheme 8 using the appropriate Intermediate (LXXXVII) in Step
8(ii). Intermediates of formula (LXXXVII) may be prepared according
to Scheme 23.
##STR00031##
[0375] wherein X is CH.sub.2 or CHMe and R.sup.4 is H or Me.
[0376] Step 23(i) An intermediate of formula (LXXXVI) may be
prepared from a commercially available lactone using hydrogenation
under palladium catalysis conditions, such as palladium on carbon
or platinum on carbon in ethyl acetate.
[0377] Step 23(ii) An intermediate of formula (LXXXVII) may be
prepared from an intermediate of formula (LXXXVI) by treatment with
boron tribromide in DCM.
[0378] In another aspect, the present invention provides a
pharmaceutical composition comprising a compound of the present
invention, or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier. The pharmaceutical composition
can be formulated for particular routes of administration such as
oral administration, parenteral administration, and rectal
administration, etc. In addition, the pharmaceutical compositions
of the present invention can be made up in a solid form (including
without limitation capsules, tablets, pills, granules, powders or
suppositories), or in a liquid form (including without limitation
solutions, suspensions or emulsions). The pharmaceutical
compositions can be subjected to conventional pharmaceutical
operations such as sterilization and/or can contain conventional
inert diluents, lubricating agents, or buffering agents, as well as
adjuvants, such as preservatives, stabilizers, wetting agents,
emulsifers and buffers, etc.
[0379] Typically, the pharmaceutical compositions are tablets or
gelatin capsules comprising the active ingredient together with
[0380] a) diluents, e.g., lactose, dextrose, sucrose, mannitol,
sorbitol, cellulose and/or glycine;
[0381] b) lubricants, e.g., silica, talcum, stearic acid, its
magnesium or calcium salt and/or polyethyleneglycol; for tablets
also
[0382] c) binders, e.g., magnesium aluminum silicate, starch paste,
gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose
and/or polyvinylpyrrolidone; if desired
[0383] d) disintegrants, e.g., starches, agar, alginic acid or its
sodium salt, or effervescent mixtures; and/or
[0384] e) absorbents, colorants, flavors and sweeteners.
[0385] Tablets may be either film coated or enteric coated
according to methods known in the art.
[0386] Suitable compositions for oral administration include an
effective amount of a compound of the invention in the form of
tablets, lozenges, aqueous or oily suspensions, dispersible powders
or granules, emulsion, hard or soft capsules, or syrups or elixirs.
Compositions intended for oral use are prepared according to any
method known in the art for the manufacture of pharmaceutical
compositions and such compositions can contain one or more agents
selected from the group consisting of sweetening agents, flavoring
agents, coloring agents and preserving agents in order to provide
pharmaceutically elegant and palatable preparations. Tablets may
contain the active ingredient in admixture with nontoxic
pharmaceutically acceptable excipients which are suitable for the
manufacture of tablets. These excipients are, for example, inert
diluents, such as calcium carbonate, sodium carbonate, lactose,
calcium phosphate or sodium phosphate; granulating and
disintegrating agents, for example, corn starch, or alginic acid;
binding agents, for example, starch, gelatin or acacia; and
lubricating agents, for example magnesium stearate, stearic acid or
talc. The tablets are uncoated or coated by known techniques to
delay disintegration and absorption in the gastrointestinal tract
and thereby provide a sustained action over a longer period. For
example, a time delay material such as glyceryl monostearate or
glyceryl distearate can be employed. Formulations for oral use can
be presented as hard gelatin capsules wherein the active ingredient
is mixed with an inert solid diluent, for example, calcium
carbonate, calcium phosphate or kaolin, or as soft gelatin capsules
wherein the active ingredient is mixed with water or an oil medium,
for example, peanut oil, liquid paraffin or olive oil.
[0387] Certain injectable compositions are aqueous isotonic
solutions or suspensions, and suppositories are advantageously
prepared from fatty emulsions or suspensions. Said compositions may
be sterilized and/or contain adjuvants, such as preserving,
stabilizing, wetting or emulsifying agents, solution promoters,
salts for regulating the osmotic pressure and/or buffers. In
addition, they may also contain other therapeutically valuable
substances. Said compositions are prepared according to
conventional mixing, granulating or coating methods, respectively,
and contain about 0.1-75%, or contain about 1-50%, of the active
ingredient.
[0388] Suitable compositions for transdermal application include an
effective amount of a compound of the invention with a suitable
carrier. Carriers suitable for transdermal delivery include
absorbable pharmacologically acceptable solvents to assist passage
through the skin of the host. For example, transdermal devices are
in the form of a bandage comprising a backing member, a reservoir
containing the compound optionally with carriers, optionally a rate
controlling barrier to deliver the compound of the skin of the host
at a controlled and predetermined rate over a prolonged period of
time, and means to secure the device to the skin.
[0389] Suitable compositions for topical application, e.g., to the
skin and eyes, include aqueous solutions, suspensions, ointments,
creams, gels or sprayable formulations, e.g., for delivery by
aerosol or the like. Such topical delivery systems will in
particular be appropriate for dermal application, e.g., for the
treatment of skin cancer, e.g., for prophylactic use in sun creams,
lotions, sprays and the like. They are thus particularly suited for
use in topical, including cosmetic, formulations well-known in the
art. Such may contain solubilizers, stabilizers, tonicity enhancing
agents, buffers and preservatives.
[0390] As used herein a topical application may also pertain to an
inhalation or to an intranasal application. They may be
conveniently delivered in the form of a dry powder (either alone,
as a mixture, for example a dry blend with lactose, or a mixed
component particle, for example with phospholipids) from a dry
powder inhaler or an aerosol spray presentation from a pressurised
container, pump, spray, atomizer or nebuliser, with or without the
use of a suitable propellant.
[0391] The present invention further provides anhydrous
pharmaceutical compositions and dosage forms comprising the
compounds of the present invention as active ingredients, since
water may facilitate the degradation of certain compounds.
[0392] Anhydrous pharmaceutical compositions and dosage forms of
the invention can be prepared using anhydrous or low moisture
containing ingredients and low moisture or low humidity conditions.
An anhydrous pharmaceutical composition may be prepared and stored
such that its anhydrous nature is maintained. Accordingly,
anhydrous compositions are packaged using materials known to
prevent exposure to water such that they can be included in
suitable formulary kits. Examples of suitable packaging include,
but are not limited to, hermetically sealed foils, plastics, unit
dose containers (e.g., vials), blister packs, and strip packs.
[0393] The invention further provides pharmaceutical compositions
and dosage forms that comprise one or more agents that reduce the
rate by which the compound of the present invention as an active
ingredient will decompose. Such agents, which are referred to
herein as "stabilizers," include, but are not limited to,
antioxidants such as ascorbic acid, pH buffers, or salt buffers,
etc.
[0394] The CFTR chloride channel is known to be associated with a
number of diseases and conditions, including cystic fibrosis (CF)
(Quinton, Physiol. Rev. 79:S3-S22 (1999); Boucher, Eur. Respir. J
23:146-58 (2004)), polycystic kidney disease (O'Sullivan et al.,
Am. J. Kidney Dis. 32:976-983 (1998); Sullivan et al., Physiol.
Rev. 78:1165-91 (1998); Strong et al., J Clin. Invest. 93:347-54
(1994); Mall et al., Gastroenterology 126:32-41 (2004); Hanaoka et
al., Am. J Physiol. 270:C389-C399 (1996); Kunzelmann et al.,
Physiol. Rev. 82:245-289 (2002); Davidow et al., Kidney Int.
50:208-18 (1996); Li et al., Kidney Int. 66:1926-38 (2004);
Al-Awqati, J Clin. Invest. 110:1599-1601 (2002); Thiagarajah et
al., Curr. Opin. Pharmacol. 3:594-99 (2003)) and secretory diarrhea
(Clarke et al., Science 257:1125-28 (1992); Gabriel et al., Science
266:107-109 (1994); Kunzelmann and Mall, Physiol. Rev. 82:245-89
(2002); Field, M. J Clin. Invest. 111:931-43 (2003); and
Thiagarajah et al., Gastroenterology 126:511-519 (2003)).
[0395] The compounds of formula (I) in free form or in salt form,
exhibit valuable pharmacological properties, e.g. CFTR modulating
properties, e.g. as indicated in in vitro and in vivo tests as
provided in the next sections, and are therefore indicated for
therapy or for use as research chemicals, e.g. as tool compounds.
The compounds of formula (I) and (Ia) may be useful in vivo in the
development of models of cystic fibrosis.
[0396] Compounds of the invention may be useful in the treatment of
an indication selected from polycystic kidney disease and diarrhea,
including infectious secretory diarrhea, travellers diarrhea,
diarrhea associated with HIV and diarrhea predominant irritable
bowel syndrome (IBS).
[0397] Thus, as a further embodiment, the present invention
provides the use of a compound of formula (I) or (Ia) in therapy.
In a further embodiment, the therapy is selected from a disease
which may be treated by inhibition of CFTR. In another embodiment,
the disease is selected from the afore-mentioned list, suitably
polycystic kidney disease and diarrhea, more suitably polycystic
kidney disease, infectious secretory diarrhea, travellers diarrhea,
diarrhea associated with HIV and diarrhea predominant irritable
bowel syndrome (IBS).
[0398] In another embodiment, the invention provides a method of
treating a disease which is treated by inhibition of CFTR
comprising administration of a therapeutically acceptable amount of
a compound of formula (I) or (Ia) In a further embodiment, the
disease is selected from the afore-mentioned list, suitably
polycystic kidney disease and diarrhea, more suitably polycystic
kidney disease, infectious secretory diarrhea, travellers diarrhea,
diarrhea associated with HIV and diarrhea predominant irritable
bowel syndrome (IBS).
[0399] Thus, as a further embodiment, the present invention
provides the use of a compound of formula (I) or (Ia) for the
manufacture of a medicament. In a further embodiment, the
medicament is for treatment of a disease which may be treated by
inhibition of CFTR. In another embodiment, the disease is selected
from the afore-mentioned list, suitably polycystic kidney disease
and diarrhea, more suitably polycystic kidney disease, infectious
secretory diarrhea, travellers diarrhea, diarrhea associated with
HIV and diarrhea predominant irritable bowel syndrome (IBS).
Embodiment 24
[0400] A compound according to any one of previous Embodiments 1 to
21, or a pharmaceutically acceptable salt thereof, for use in the
treatment of polycystic kidney disease and diarrhea, more suitably
polycystic kidney disease, infectious secretory diarrhea,
travellers diarrhea, diarrhea associated with HIV and diarrhea
predominant irritable bowel syndrome (IBS).
[0401] The pharmaceutical composition or combination of the present
invention can be in unit dosage of about 1-2000 mg of active
ingredient(s) for a subject of about 50-70 kg, or about 1-1000 mg
or about 1-500 mg or about 1-250 mg or about 1-150 mg or about
0.5-100 mg, or about 1-50 mg of active ingredients. The
therapeutically effective dosage of a compound, the pharmaceutical
composition, or the combinations thereof, is dependent on the
species of the subject, the body weight, age and individual
condition, the disorder or disease or the severity thereof being
treated. A physician, clinician or veterinarian of ordinary skill
can readily determine the effective amount of each of the active
ingredients necessary to prevent, treat or inhibit the progress of
the disorder or disease.
[0402] The above-cited dosage properties are demonstrable in vitro
and in vivo tests using advantageously mammals, e.g., mice, rats,
dogs, monkeys or isolated organs, tissues and preparations thereof.
The compounds of the present invention can be applied in vitro in
the form of solutions, e.g., aqueous solutions, and in vivo either
enterally, parenterally, advantageously intravenously, e.g., as a
suspension or in aqueous solution. The dosage in vitro may range
between about 10.sup.-3 molar and 10.sup.-9 molar concentrations. A
therapeutically effective amount in vivo may range depending on the
route of administration, between about 0.1-500 mg/kg, or between
about 1-100 mg/kg.
[0403] The activity of a compound according to the present
invention can be assessed by the following methods.
IonWorks Quattro assay:
[0404] CFTR activity can be quantified by electrophysiology
methods, using the whole-cell configuration of the patch clamp
technique (Hamill O, Marty A, Neher E, Sakmann B and Sigworth F.
`Improved patch-clamp techniques for high resolution current
recording from cells and cell-free membrane patches.` Pflugers
Archive 1981 391: 85-100). This assay directly measures the
currents associated with chloride flow through CFTR channels whilst
either maintaining or adjusting the transmembrane voltage. This
assay can use either single glass micropipettes or parallel planar
arrays to measure CFTR activity from native or recombinant cell
systems. Currents measured using parallel planar arrays can be
quantified using an appropriately equipped instrument such as the
IonWorks Quattro (Molecular Devices Corporation, Sunnyvale,
Calif.). The Quattro system can measure CFTR currents from either a
single cell per recording well (HT configuration) or alternatively
from a population of 64 cells per well (Population Patch Clamp PPC)
(Finkel A, Wittel A, Yang N, Handran S, Hughes J, Costantin J.
`Population patch clamp improves data consistency and success rates
in the measurement of ionic currents.` J Biomol Screen. 2006
August; 11 (5):488-96).
Cell Culture:
[0405] Chinese hamster ovary (CHO) cells stably expressing WT-CFTR
channels were used for the IonWorks Quattro experiments. Cells were
maintained at 37.degree. C. in 5% v/v CO.sub.2 at 100% humidity in
MEM alpha medium supplemented with 10% (v/v) FCS, 100 U/mL
Penicillin/Streptomycin, and 100 .mu.g/L methotrexate. For
experiments, cells were grown in 225 cm.sup.2 tissue culture flasks
until almost confluent. Cells were removed from the flask using
trypsin-EDTA and resuspended in extracellular recording solution
for immediate experimentation.
CFTR Inhibitor Assay:
[0406] Cells, at a density of 1.5-2 million per mL, were placed on
the Quattro system, added to the planar patch plate and seals
allowed to establish for 5-10 mins. After assessing seal
resistances (typically >50 M.OMEGA.), whole-cell access was
obtained by perforation with 100 .mu.g/mL amphotericin B. Baseline
currents were measured by a pre-compound scan obtained by
application of a voltage ramp from -100 to +100 mV. CFTR was
activated by the addition of 10 .mu.M forskolin to each of the 384
wells of the patch plate. After 5 min incubation the post-compound
1 currents were measured, again by application of a voltage ramp
from -100 to +100 mV. Test compounds, diluted from 10 mM stocks in
DMSO in extracellular solution were then added to the patch plate
and were incubated for a further 10 min. The post-compound 2
currents were measured by the application of the same voltage ramp
from -100 to +100 mV. The inhibition of CFTR was determined from
the difference in current between the forskolin addition
(post-compound 1) and the test compound (post-compound 2). This was
determined at both -100 mV and +100 mV which represents the inward
and outward current respectively.
Solutions:
[0407] Extracellular solution (ECS): 145 mM NaCl, 4 mM CsCl, 5 mM
D-glucose, 10 mM TES, 1 mM CaCl.sub.2, 1 mM MgCl.sub.2, pH 7.4
NaOH
[0408] Intracellular solution (ICS): 113 mM L-Aspartic acid, 113 mM
CsOH, 27 mM CsCl, 1 mM NaCl, 1 mM MgCl.sub.2, 1 mM EGTA, 10 mM TES.
pH 7.2 with CsOH. Filter sterilized before use.
[0409] Using the Ion Works Quattro assay (as described in this
application) compounds of the invention exhibit CFTR inhibitory
efficacy in accordance with Table A:
TABLE-US-00001 TABLE A Inhibitory Activity of Compounds of Formula
(I) CFTR IC50 (.mu.M) Example Number Inward current 1.2 0.79 1.3
>30 1.1.1 0.63 1.6 11.13 1.10 8.08 2.1 0.41 3.1.1 0.41 3.1.2
1.51 3.5 0.16 4.2 1.67 6.1 27.63 6.3 2.24 6.4 >30 7.5b 2.18 8.2
0.29 9.1 0.29 9.2 9.30 10.0 2.15 10.2 0.70 11.0 9.14 12.1b 0.18
12.b 0.54 12.a 0.20 13.1a 0.10 13.2a 0.05 13a 0.07 13.3a 0.09 14a
0.31 15 0.10 15.7 5.50 15.8 3.93 15.9 1.90 15.10 1.18 15.11 0.13
15.12 9.27 15.13 0.62 17 0.10 17.1 0.23 17.2 0.24 18 0.48 19 5.38
20a 0.27 21a 0.33 21.8 1.39 22 0.47 23a 0.05 23.1a 0.10 24 1.74 25
4.84 26a 0.15 26.1a 0.23 26.3a 0.45 27a 0.15 28a 0.11 29 0.44 30a
0.16 30.1a 0.25 31 1.45
[0410] The compounds of the present invention may be administered
either simultaneously with, or before or after, one or more other
therapeutic agent. The compound of the present invention may be
administered separately, by the same or different route of
administration, or together in the same pharmaceutical composition
as the other agents.
[0411] In one embodiment, the invention provides a product
comprising a compound of formula (I) and at least one other
therapeutic agent as a combined preparation for simultaneous,
separate or sequential use in therapy. In one embodiment, the
therapy is the treatment of a disease or condition mediated by
CFTR. Products provided as a combined preparation include a
composition comprising the compound of formula (I) and the other
therapeutic agent(s) together in the same pharmaceutical
composition, or the compound of formula (I) and the other
therapeutic agent(s) in separate form, e.g. in the form of a
kit.
[0412] In one embodiment, the invention provides a pharmaceutical
composition comprising a compound of formula (I) and another
therapeutic agent(s). Optionally, the pharmaceutical composition
may comprise a pharmaceutically acceptable carrier, as described
above.
[0413] In one embodiment, the invention provides a kit comprising
two or more separate pharmaceutical compositions, at least one of
which contains a compound of formula (I). In one embodiment, the
kit comprises means for separately retaining said compositions,
such as a container, divided bottle, or divided foil packet. An
example of such a kit is a blister pack, as typically used for the
packaging of tablets, capsules and the like.
[0414] The kit of the invention may be used for administering
different dosage forms, for example, oral and parenteral, for
administering the separate compositions at different dosage
intervals, or for titrating the separate compositions against one
another. To assist compliance, the kit of the invention typically
comprises directions for administration.
[0415] In the combination therapies of the invention, the compound
of the invention and the other therapeutic agent may be
manufactured and/or formulated by the same or different
manufacturers. Moreover, the compound of the invention and the
other therapeutic may be brought together into a combination
therapy: (i) prior to release of the combination product to
physicians (e.g. in the case of a kit comprising the compound of
the invention and the other therapeutic agent); (ii) by the
physician themselves (or under the guidance of the physician)
shortly before administration; (iii) in the patient themselves,
e.g. during sequential administration of the compound of the
invention and the other therapeutic agent.
[0416] Accordingly, the invention provides the use of a compound of
formula (I) or (Ia) for treating a disease or condition mediated by
CFTR, wherein the medicament is prepared for administration with
another therapeutic agent. The invention also provides the use of
another therapeutic agent for treating a disease or condition
mediated by CFTR, wherein the medicament is administered with a
compound of formula (I).
[0417] The invention also provides a compound of formula (I) or
(Ia) for use in a method of treating a disease or condition
mediated by CFTR, wherein the compound of formula (I) or (Ia) is
prepared for administration with another therapeutic agent. The
invention also provides another therapeutic agent for use in a
method of treating a disease or condition mediated by CFTR, wherein
the other therapeutic agent is prepared for administration with a
compound of formula (I) or (Ia). The invention also provides a
compound of formula (I) or (Ia) for use in a method of treating a
disease or condition mediated by CFTR, wherein the compound of
formula (I) or (Ia) is administered with another therapeutic agent.
The invention also provides another therapeutic agent for use in a
method of treating a disease or condition mediated by CFTR, wherein
the other therapeutic agent is administered with a compound of
formula (I) or (Ia).
[0418] The invention also provides the use of a compound of formula
(I) for treating a disease or condition mediated by CFTR, wherein
the patient has previously (e.g. within 24 hours) been treated with
another therapeutic agent. The invention also provides the use of
another therapeutic agent for treating a disease or condition
mediated by CFTR, wherein the patient has previously (e.g. within
24 hours) been treated with a compound of formula (I).
[0419] In one embodiment, the invention provides a combination
comprising a therapeutically effective amount of a compound of
formula (I) or (Ia) according to any preceding Embodiment, or a
pharmaceutically acceptable salt thereof, and one or more
therapeutically active co-agent is selected from an anti-diarrheal
agent, including an oral rehydration agent; an antibiotic; and an
antimotility agent such as loperamide.
[0420] Specific individual combinations which may provide
particular treatment benefits include a combination of a compound
of formula (I) or (Ia) according to any preceding Embodiment, or a
pharmaceutically acceptable salt thereof, and loperamide.
EXAMPLES
General Conditions
[0421] Mass spectra were acquired on LC-MS, SFC-MS, or GC-MS
systems using electrospray, chemical and electron impact ionization
methods from a range of instruments of the following
configurations: Agilent 1100 HPLC systems with an Agilent 6110 Mass
Spectrometer, or Micromass Platform Mass Spectrometer or Thermo LTQ
Mass Spectrometer; a Waters Acquity UPLC system with SQD Mass
Spectrometer, a Waters FractionLynx HPLC system with 3100 Mass
Spectrometer, a Waters UPC2 system with TQD Mass Spectrometer or a
Waters Prep100 SFC-MS system with SQD2 Mass Spectrometer. [M+H]+
refers to protonated molecular ion of the chemical species. NMR
spectra were run on Bruker AVANCE 400 MHz or 500 MHz NMR
spectrometers using ICON-NMR, under TopSpin program control.
Spectra were measured at 298K, unless indicated otherwise, and were
referenced relative to the solvent resonance.
[0422] The following examples are intended to illustrate the
invention and are not to be construed as being limitations thereon.
Temperatures are given in degrees centigrade.
[0423] If not mentioned otherwise, all evaporations are performed
under reduced pressure, preferably between about 15 mm Hg and 100
mm Hg (=20-133 mbar). The structure of final products,
intermediates and starting materials is confirmed by standard
analytical methods, e.g., microanalysis and spectroscopic
characteristics, e.g., MS, IR, NMR.
[0424] Abbreviations used are those conventional in the art. If not
defined, the terms have their generally accepted meanings.
ABBREVIATIONS
[0425] BOC tertiary butyl carboxy br broad CBA weak cation exchange
(e.g Isolute.RTM. CBA columns from Biotage) d doublet d.e. or de
diastereomeric excess
deg C. .degree. C.
[0426] dd doublet of doublets ddd doublet of doublet of doublets
DBU 1,8-Diazabicyclo[5.4.0]undec-7-ene DCM dichloromethane DIPEA
diisopropylethylamine DMA dimethyl acetamide DMAP
4-dimethylaminopyridine
DMF N,N-dimethylformamide
[0427] DMSO dimethylsulfoxide e.e. or ee enantiomeric excess EtOAc
ethyl acetate EtOH ethanol h hour(s) HCl hydrochloric acid HPLC
high pressure liquid chromatography IR infra-red KOtBu potassium
t-butoxide LCMS liquid chromatography and mass spectrometry LDA
lithium diisopropylamide MeOH methanol 2-Me-THF
2-methyltetrahydrofuran MS mass spectrometry m multiplet mult
multiplet min minutes mL or ml milliliter(s) m/z mass to charge
ratio NaOH sodium hydroxide
NBS N-bromosuccinimide
[0428] NMR nuclear magnetic resonance Pd-118
1,1''bis(di-t-butylphosphino)ferrocene palladium dichloride ppm
parts per million PS polymer supported RT room temperature Rt
retention time s singlet SCX-2 strong cation exchange (e.g.
Isolute.RTM. SCX-2 columns from Biotage) SEM-Cl
(2-(chloromethoxy)ethyl)trimethylsilane SFC Supercritical fluid
chromotography t triplet T3P.RTM.
2,4,6-Tripropyl-1,3,5,2,4,6-trioxatriphosphorinane-2,4,6-trioxide
solution TBAF Tetrabutylammonium fluoride solution TBS-Cl
tert-Butyldimethylsilyl chloride TEA triethylamine TFA
trifluoroacetic acid THF tetrahydrofuran
[0429] Referring to the examples that follow, compounds of the
preferred embodiments were synthesized using the methods described
herein, or other methods, which are known in the art.
[0430] The various starting materials, intermediates, and compounds
of the preferred embodiments may be isolated and purified, where
appropriate, using conventional techniques such as precipitation,
filtration, crystallization, evaporation, distillation, and
chromatography. Unless otherwise stated, all starting materials are
obtained from commercial suppliers and used without further
purification. Salts may be prepared from compounds by known
salt-forming procedures.
[0431] It should be understood that the organic compounds according
to the preferred embodiments may exhibit the phenomenon of
tautomerism. As the chemical structures within this specification
can only represent one of the possible tautomeric forms, it should
be understood that the preferred embodiments encompasses any
tautomeric form of the drawn structure.
[0432] If not indicated otherwise, the analytical HPLC conditions
are as follows:
2minLC_v003
TABLE-US-00002 Column Waters BEH C18 50 .times. 2.1 mm, 1.7 .mu.m
Column Temperature 50.degree. C. Eluents A: H.sub.2O, B:
acetonitrile, both containing 0.1% TFA Flow Rate 0.8 mL/min
Gradient 0.20 min 5% B; 5% to 95% B in 1.30 min, 0.25 min 95% B
10minLC_v003
TABLE-US-00003 Column Waters BEH C18 50 .times. 2.1 mm, 1.7 .mu.m
Column 50.degree. C. Temperature Eluents A: H.sub.2O, B:
acetonitrile, both containing 0.1% TFA Flow Rate 0.8 mL/min
Gradient 0.20 min 5% B; 5% to 95% B in 7.80 min, 1.00 min 95% B
2minLowpH:
TABLE-US-00004 Column: Waters Acquity CSH 1.7 .mu.m, 2.1 .times. 50
mm Temperature: 50.degree. C. Mobile Phase: A: Water + 0.1% Formic
Acid B: Acetonitrile + 0.1% Formic Acid Flow rate: 1.0 mL/min
Gradient: 0.0 min 5% B, 0.2-1.3 min 5-98% B, 1.3-1.55 min 98% B,
1.55-1.6 min 98-5% B
2minLowpHv01:
TABLE-US-00005 Column: Waters Acquity CSH 1.7 .mu.m, 2.1 .times. 50
mm Temperature: 50.degree. C. Mobile Phase: A: Water + 0.1% Formic
Acid B: Acetonitrile + 0.1% Formic Acid Flow rate: 1.0 mL/min
Gradient: 0.0 min 5% B, 0.2-1.55 min 5-98% B, 1.55-1.75 min 98% B,
1.75-1.8 min 98-5% B
2minLowpHv02:
TABLE-US-00006 Column: Waters Acquity CSH 1.7 .mu.m, 2.1 .times. 50
mm Temperature: 50.degree. C. Mobile Phase: A: Water + 0.1% TFA B:
Acetonitrile + 0.1% TFA Flow rate: 1.0 mL/min Gradient: 0.0 min 5%
B, 0.2-1.55 min 5-98% B, 1.55-1.75 min 98% B, 1.75-1.8 min 98-5%
B
2minLowpHv03:
TABLE-US-00007 Column: Waters Acquity CSH 1.7 .mu.m, 2.1 .times. 50
mm Temperature: 50.degree. C. Mobile Phase: A: Water + 0.1% Formic
Acid B: Acetonitrile + 0.1% Formic Acid Flow rate: 1.0 mL/min
Gradient: 0.0 min 5% B, 0.2-1.8 min 5-98% B, 1.8-2.1 min 98% B,
2.1-2.3 min 98% B
2minLowpHv04:
TABLE-US-00008 Column: Waters Acquity CSH C18 50 .times. 2.1 mm
Temperature: 50.degree. C. Mobile Phase: A: Water + 0.1% TFA B:
Acetonitrile + 0.1% TFA Flow rate: 1.0 mL/min Gradient: 0.0 min 5%
B, 0.2-1.8 min 5-98% B, 1.8-2.1 min 98% B
Preparation of Final Compounds
[0433] Unless otherwise indicated, the chemical name refers to the
racemic compound or to the diastereomeric mixture. The term
`Enantiomer 1` refers to the more active of the separated
enantiomers and does not indicate the relative position of the
eluted peak within the chromatogram (where separation is achieved
by SFC under the specified conditions). The term `Diastereomer 1`
refers to the more active of the separated diastereomers and does
not indicate the relative position of the eluted peak within the
chromatogram (where separation is achieved by SFC under the
specified conditions).
Example 1.1
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[4',5-
':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00032##
[0435] To
6-(2-mercapto-ethyl)-1,3-dimethyl-5-phenyl-1,6-dihydro-pyrrolo[3-
,4-d]pyrimidine-2,4-dione (Intermediate A) (207 mg, 0.656 mmol) and
bismuth triflate (43.1 mg, 0.066 mmol) in toluene (5 ml) was added
5-methylfuran-2-carbaldehyde (commercial) (0.065 ml, 0.656 mmol)
the mixture heated to 100.degree. C. for 1 hour. The reaction
mixture was cooled to RT and the solvent was removed under reduced
pressure. The residue was partitioned between EtOAc (20 mL) and
water (20 mL) and the phases separated. The aqueous phase was
extracted with EtOAc (2.times.20 ml) the combined organic phases
were dried over magnesium sulfate and the solvent removed under
vacuum. Purification by chromatography on silica, eluting with
0-50% EtOAc/hexane gave the title compound. .sup.1H NMR (400 MHz,
DMSO-d6) .delta. 7.45 (5H, s), 6.03 (1H, s), 5.99 (1H, dd), 5.88
(1H, d), 4.18-4.11 (1H, m), 4.02-3.97 (1H, m), 3.42 (3H, s), 3.11
(3H, s), 3.03-2.89 (2H, m), 2.25 (3H, s).
[0436] LC-MS: Rt=1.11 mins; MS m/z 408.4 [M+H].sup.+ Method
2minLowpH
[0437] Chiral separation of the racemate by Supercritical Fluid
Chromatography was carried out using the following conditions to
afford the compounds listed hereinafter:
[0438] Mobile Phase: 50% MeOH/50% CO2
[0439] Column: Chiralcel OJ-H, 250.times.10 mm, 5 um
[0440] Detection: UV @ 220 nm
[0441] Flow rate: 10 mL/min
[0442] Injection volume: 200 .mu.l
Examples 1.2 and 1.3 are Enantiomers.
Example 1.2
Enantiomer 1 of
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[4',-
5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[0443] SFC retention time=5.09 min.
[0444] 1HNMR (400 MHz, DMSO-d6) .delta. 7.45 (5H, s), 6.03 (1H, s),
5.99 (1H, dd), 5.88 (1H, d), 4.18-4.11 (1H, m), 4.02-3.97 (1H, m),
3.42 (3H, s), 3.11 (3H, s), 3.03-2.89 (2H, m), 2.25 (3H, s).
[0445] LC-MS: Rt=1.11 mins; MS m/z 408.4 [M+H].sup.+ Method
2minLowpH Chiral purity 99% ee
Example 1.3
Enantiomer 2 of
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8-dihydro-1H-pyrimido[4',-
5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[0446] SFC retention time=6.82 min.
[0447] 1H NMR (400 MHz, DMSO-d6) .delta. 7.45 (5H, s), 6.03 (1H,
s), 5.99 (1H, dd), 5.88 (1H, d), 4.18-4.11 (1H, m), 4.02-3.97 (1H,
m), 3.42 (3H, s), 3.11 (3H, s), 3.03-2.89 (2H, m), 2.25 (3H,
s).
[0448] LC-MS: Rt=1.11 mins; MS m/z 408.4 [M+H].sup.+ Method
2minLowpH Chiral purity 99% ee
[0449] The compounds of the following tabulated examples (Table 1)
were prepared by a similar method to that of Example 1.1 from the
appropriate thiol (preparation described hereinafter) and
commercially available aldehyde.
TABLE-US-00009 TABLE 1 Ex Structure Name LC-MS NMR 1.4 ##STR00033##
1,3-Dimethyl-10-(4- methylthiazol-2-yl)- 5-phenyl-7,8- dihydro-1H-
pyrimido[4',5':3,4] pyrrolo[2,1- c][1,4]thiazine- 2,4(3H,10H)-dione
LC-MS: Rt 1.03 mins; 425.4 [M + H]+; 2 minLowpH 1H NMR (400 MHz,
CDCl3) .delta. 7.54-7.43 (m, 5H), 6.86 (d, 1H), 6.02 (s, 1H), 4.21
(dd, 1H), 4.20 (dd, 1H), 3.64 (s, 3H), 3.36 (s, 3H), 3.21 (ddd,
1H), 2.90 (ddd, 1H), 2.46 (d, 3H) 1.5 ##STR00034##
10-(5-Chlorofuran- 2-yl)-1,3-dimethyl- 5-phenyl-7,8- dihydro-1H-
pyrimido[4',5':3,4] pyrrolo[2,1- c][1,4]thiazine- 2,4(3H,10H)-dione
LC-MS: Rt 1.13 mins; 428 [M + H]+; 2 minLowpH; 1H NMR: (400 MHz,
CDCl3) .delta. 7.53-7.46 (3H, m), 7.44-7.41 (2H, m), 6.10 (1H, d),
5.94 (1H, dd), 5.73 (1H, s), 4.24- 4.17 (1H, m), 4.16- 4.08 (1H,
m), 3.59 (3H, s), 3.35 (3H, s), 3.11-3.04 (1H, m), 2.87-2.80 (1H,
m). 1.6 ##STR00035## 10-(3- Chlorophenyl)-1,3- dimethyl-5-phenyl-
7,8-dihydro-1H- pyrimido[4',5':3,4] pyrrolo[2,1- c][1,4]thiazine-
2,4(3H,10H)-dione LC-MS: Rt 1.12 mins; 438 [M + H]+; 2 minLowpH; 1H
NMR: (400 MHz, CDCl3) .delta. 7.54-7.44 (5H, m), 7.28 (2H, m), 7.17
(1H, s), 6.99 (1H, d), 5.73 (1H, s), 4.17-4.06 (2H, m), 3.46 (3H,
s), 3.35 (3H, s), 2.98-2.90 (1H, m), 2.80-2.73 (1H, m) 1.7
##STR00036## 1,3-Dimethyl-5- phenyl-10-(m-tolyl)- 7,8-dihydro-1H-
pyrimido[4',5':3,4] pyrrolo[2,1- c][1,4]thiazine- 2,4(3H,10H)-dione
LC-MS: Rt 1.16 mins; 418.5 [M + H]+; 2 minLowpH 1H NMR: (400 MHz,
CDCl3) .delta. 7.49 (5H, m), 7.22 (1H, t), 7.09 (1H, d), 6.99 (1H,
s), 6.86 (1H, d), 5.75 (1H, s), 4.12 (2H, m), 3.46 (3H, s), 3.35
(3H, s), 2.98 (1H, m), 2.75 (1H, m), 2.37 (3H, s). 1.8 ##STR00037##
10-(5-Chlorofuran- 2-yl)-1,3-dimethyl- 5-(m-tolyl)-7,8- dihydro-1H-
pyrimido[4',5':3,4] pyrrolo[2,1- c][1,4]thiazine- 2,4(3H,10H)-dione
LC-MS: Rt 1.14 mins; 442.4 [M + H]+; 2 minLowpH; 1H NMR: (400 MHz,
DMSO-d6) .delta. 7.35 (1H, q), 7.29-7.22 (3H, m), 6.41 (1H, d),
6.13 (1H, s), 6.11 (1H, dd), 4.13-4.07 (1H, m), 3.99-3.92 (1H, m),
3.44 (3H, s), 3.13 (3H, s), 2.96-2.92 (2H, m), 2.36 (3H, s). 1.9
##STR00038## 1,3-Dimethyl-10-(4- methylthiazol-2-yl)-
5-(m-tolyl)-7,8- dihydro-1H- pyrimido[4',5':3,4] pyrrolo[2,1-
c][1,4]thiazine- 2,4(3H,10H)-dione LC-MS; Rt 1.09 mins; 439.4 [M +
H]+; 2 minLowpH; 1H NMR (400 MHz, DMSO-d6) .delta. 7.36 (1H, t),
7.29-7.19 (4H, m), 6.36 (1H, s), 4.11-3.99 (1H, m), 3.52 (3H, s),
3.18 (3H, s), 3.11 (1H, m), 3.02 (1H, m), 2.35 (3H, s), 2.30 (3H,
s) 1.10 ##STR00039## 1,3-Dimethyl-10-(5- methylthiophen-2-
yl)-5-phenyl-7,8- dihydro-1H- pyrimido[4',5':3,4] pyrrolo[2,1-
c][1,4]thiazine- 2,4(3H,10H)-dione LC-MS Rt 1.18 mins [M + H].sup.+
424.2 (Method 2 minLowpH) 1H NMR (400 MHz, CDCl3) .delta. 7.47 (5H,
mult), 6.86 (1H, s), 6.50 (1H, s), 5.87 (1H, s), 4.11 (2H, mult),
3.62 (3H, s), 3.37 (3H, s), 3.16 (1H, mult), 2.87 (1H, mult), 2.21
(3H, s). 1.11 ##STR00040## 1,3-Dimethyl-10-(4- methylthiophen-2-
yl)-5-phenyl-7,8- dihydro-1H- pyrimido[4',5':3,4] pyrrolo[2,1-
c][1,4]thiazine- 2,4(3H,10H)-dione LC-MS Rt 1.30 mins [M + H].sup.+
424.3 (Method 2 minLowpHv01) 1H NMR (400 MHz, CDCl3) .delta. 7.47
(5H, mult), 6.86 (1H, s), 6.51 (1H, s), 5.87 (1H, s), 4.11 (2H,
mult), 3.62 (3H, s), 3.37 (3H, s), 3.16 (1H, mult), 2.87 (1H,
mult), 2.21 (3H, s)
[0450] The compounds of the following tabulated Examples (Table
1.1) were prepared by SFC chromatographic resolution of the
appropriate racemate.
TABLE-US-00010 TABLE 1.1 LC-MS NMR Structure SFC (Method; Ex Name
Rt (min); Chiral purity) 1.1.1 ##STR00041##
(R)-10-(5-chlorofuran-2-yl)-1,3- dimethyl-5-phenyl-7,8-dihydro-1H-
pyrimido[4',5':3,4]pyrrolo[2,1- c][1,4]thiazine-2,4(3H,10H)-dione
(see X-ray data below table) LC-MS: Rt 1.14 mins; 428 [M + H]+; 2
minLowpH; 1H NMR: (400 MHz, DMSO-d6) .delta. 7.47 (5H, s), 6.41
(1H, d), 6.14 (1H, s), 6.13 (1H, s), 4.14-4.07 (1H, m), 4.02-3.95
(1H, m), 3.45 (3H, s), 3.14 (3H, s), 2.97-2.93 (2H, m). SFC:
Co-SolventlPA + 0.1% DEA Instrument Method OJ50IPA_DEA Column
CHIRALCEL OJ-H 250 .times. 10 mm, 5 mic Rt 5.32 min; >99.9% ee
1.1.2 ##STR00042## Enantiomer 1 of
1,3-Dimethyl-10-(4-methylthiazol-2- yl)-5-phenyl-7,8-dihydro-1H-
pyrimido[4',5':3,4]pyrrolo[2,1- c][1,4]thiazine-2,4(3H,10H)-dione
LC-MS: Rt 1.04 mins; 425.2 [M + H]+; 2 minLowpH; 1H NMR: (400 MHz,
DMSO-d6) .delta. 7.47 (5H, m), 7.25 (1H, s), 6.39 (1H, s),
4.15-4.08 (1H, m), 4.09- 3.99 (1H, m), 3.53 (3H, s), 3.15 (3H, s),
3.13-3.08 (1H, m), 3.04- 2.99 (1H, m), 2.31 (3H, s). SFC: Column:
Chiralcel OJ-H 250 .times. 10 mm, 5 um Mobile phase: 50% methanol +
0.1% DEA/50% CO2 Flow: 10 ml/min Detection: UV @ 220 nm Rt 4.99
min; >99% ee 1.1.3 ##STR00043## Enantiomer 1 of
1,3-Dimethyl-10-(5-methylfuran-2- yl)-5-phenyl-7,8-dihydro-1H-
pyrimido[4',5':3,4]pyrrolo[2,1- c][1,4]thiazine-2,4(3H,10H)-dione
LC-MS: Rt 1.12 min; 408.2 [M + H]+; 2 minLowpH; 1H NMR: (400 MHz,
DMSO-d6) .delta. 7.42 (5H, s), 6.05 (1H, s), 5.98 (1H, d), 5.86
(1H, d), 5.16-4.10 (1H, m), 4.02-3.94 (1H, m), 3.44 (3H, s), 3.13
(3H, s), 3.01-2.87 (2H, m), 2.26 (3H, s). SFC: Column: Chiralcel
OJ-H 250 .times. 10 mm, 5 um Mobile phase: 50% methanol/50% CO2
Flow: 10 ml/min Detection: UV @ 220 nm Column temp: 35 deg C Berger
Minigram system 2 Rt 5.09 min; >99.9% ee 1.1.4 ##STR00044##
Enantiomer 1 of 10-(3-Chlorophenyl)-1,3-dimethyl-
5-phenyl-7,8-dihydro-1H- pyrimido[4',5':3,4]pyrrolo[2,1-
c][1,4]thiazine-2,4(3H,10H)-dione LC-MS: Rt 1.19 mins; 438.2 [M +
H]+; 2 minLowpH; 1H NMR: (400 MHz, DMSO-d6) .delta. 7.52-7.45 (5H,
m), 7.39-7.30 (3H, m), 7.00 (1H, d), 6.17 (1H, s), 4.15 (1H, dt),
3.98-3.90 (1H, m), 3.30 (3H, s), 3.12 (3H, s), 2.91- 2.75 (2H, m)
SFC: Column: Chiralcel OD-H 250 .times. 10 mm, 5 um @ 35degC Mobile
phase: 50% Isopropanol/ 50% CO2 Flow: 10 ml/min Detection: UV @ 220
nm System: Berger Minigram SFC1 Rt 7.88 min; >99.9% ee 1.1.5
##STR00045## Enantiomer 1 of 10-(5-Chlorofuran-2-yl)-1,3-
dimethyl-5-(m-tolyl)-7,8-dihydro-
1H-pyrimido[4',5':3,4]pyrrolo[2,1-
c][1,4]thiazine-2,4(3H,10H)-dione LC-MS: Rt 1.19 mins; 442.4 [M +
H]+; 2 minLowpH; 1H NMR: (400 MHz, DMSO-d6) .delta. 7.35 (1H, q),
7.29-7.22 (3H, m), 6.41 (1H, d), 6.13 (1H, s), 6.11 (1H, dd),
4.13-4.07 (1H, m), 3.99-3.92 (1H, m), 3.44 (3H, s), 3.13 (3H, s),
2.96-2.92 (2H, m), 2.36 (3H, s). SFC: Column: Chiralcel OD-H 250
.times. 10 mm, 5 um Mobile phase: 40% methanol + 0.1% DEA/60% CO2
Flow: 10 ml/min Detection: UV @ 220 nm Rt 7.34 min; >99.9%
ee
Example 1.1.1
X-ray Crystallography
[0451] Absolute stereochemistry of
(R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido-
[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione was
confirmed by X-ray crystallography:
##STR00046##
TABLE-US-00011 TABLE 1 Crystal data and structure refinement
Empirical formula C21H18ClN3O3S Formula weight 427.89 Temperature
100(2) K Wavelength 1.54178 .ANG. Crystal system Orthorhombic Space
group P212121 Unit cell dimensions a = 7.1030(10) .ANG. .alpha. =
90.degree. b = 11.330(2) .ANG. .beta. = 90.degree. c = 23.851(4)
.ANG. .gamma. = 90.degree. Volume 1919.5(5) .ANG..sup.3 Z 4 Density
(calculated) 1.481 g/cm.sup.3 Absorption coefficient 3.030
mm.sup.-1 F(000) 888 Crystal size 0.31 .times. 0.14 .times. 0.03
mm.sup.3 Theta range for data collection 3.71 to 68.36.degree.
Index ranges -8 <= h <= 8, -13 <= k <= 13, -28 <= l
<= 28 Reflections collected 40374 Independent reflections 3517
[R(int) = 0.0389] Completeness to theta = 68.36.degree. 99.9%
Absorption correction Semi-empirical from equivalents Max. and min.
transmission 0.9146 and 0.4535 Refinement method Full-matrix
least-squares on F.sup.2 Data/restraints/parameters 3517/68/273
Goodness-of-fit on F.sup.2 1.040 Final R indices [I > 2sigma(I)]
R1 = 0.0240, wR2 = 0.0635 R indices (all data) R1 = 0.0243, wR2 =
0.0637 Absolute structure parameter 0.010(10) Largest diff. peak
and hole 0.190 and -0.225 e .ANG..sup.-3
Example 2.0
7-((dimethylamino)methyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one
##STR00047##
[0453]
(S)-6-(1-(dimethylamino)-3-mercaptopropan-2-yl)-1,3-dimethyl-5-phen-
yl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (Intermediate B)
(370 mg, 0.993 mmol), 5-methylfurfural (0.109 mL, 1.093 mmol) and
bismuth triflate (65.2 mg, 0.099 mmol) were combined in toluene (15
mL) and the mixture was heated at 100.degree. C. for 1 hour 40mins.
The reaction mixture was cooled to room temperature and evaporated
under vacuum. The residue was partitioned between DCM and sat.
NaHCO.sub.3(aq) and the phases were separated. The organic phase
was passed through a hydrophobic frit and evaporated under vacuum.
Purification by chromatography on silica, eluting with 10-50%
EtOAc/hexane afforded a diastereomeric mixture of
7-((dimethylamino)methyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7-
,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-d-
ione (Example 2.0).
[0454] The diastereoisomers were separated under the following
conditions to afford the compound listed hereinafter:
[0455] Mobile Phase: 20% (MeOH+0.1% DEA)/80% CO.sub.2
[0456] Column: Chiralcel OJ-H 250 mm.times.10 mm.times.5 .mu.m
[0457] Detection: UV @ 220 nm
[0458] Flow rate: 10 mL/min
Example 2.1
Diastereomer 1 of
7-((dimethylamino)methyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7-
,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-d-
ione
[0459] SFC Retention Time=8.42 min (second eluted peak)
[0460] LC-MS: Rt 0.75 mins; MS 465.6 m/z [M+H] Method
2minLowpHv01
[0461] 1H NMR (400 MHz, CDCl3) .delta. 7.64-7.33 (5H, mult),
5.96-5.83 (3H, mult), 4.58 (1H, dddd), 3.55 (3H, s), 3.34 (3H, s),
3.18 (1H, dd), 3.13 (1H, dd), 2.77 (1H, mult), 2.32 (3H, s), 2.22
(1H, dd), 1.89 (6H, s).
Example 3.0
(8S)-10-(5-chlorofuran-2-yl)-1,3,8-trimethyl-5-phenyl-7,8-dihydro-1H-pyrim-
ido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00048##
[0463]
(S)-6-(2-Mercaptopropyl)-1,3-dimethyl-5-phenyl-1H-pyrrolo[3,4-d]pyr-
imidine-2,4(3H,6H)-dione (Intermediate E) (200 mg 0.61 mmol) was
added to a microwave vial (0.5-2 mL) equipped with a stirrer bar.
To the vial was added toluene (2 ml) followed by bismuth triflate
(38 mg, 0.061 mmol) and 5-chlorofuran-2-carbaldehyde (87 mg, 0.67
mmol). The reaction mixture was heated to 100.degree. C. in a
microwave reactor for 15 min and then reduced in vacuo. The
resulting residue was partitioned between EtOAc and water and the
layers were separated. The aqueous was extracted with EtOAc and the
combined organic extracts were dried (MgSO.sub.4), filtered and
concentrated in vacuo to yield an oil. The brown oil was dissolved
in smallest minimal volume of DCM and purified by chromatography on
silica eluting with 0-40% EtOAc in iso-hexane to afford a
diasteromeric mixture of
(8S)-10-(5-chlorofuran-2-yl)-1,3,8-trimethyl-5-phenyl-7,8-dihydro-1H-p-
yrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione as
a pale yellow white solid. LCMS Rt 1.32 mins; MS m/z 442 [M+H]+;
(Method 2minLowpHv01). The diastereoisomers were separated under
the following conditions to afford the compounds listed
hereinafter:
[0464] Mobile phase: 50% Isopropanol/50% CO2
[0465] Column: Chiralpak IB, 250.times.10 mm, 5 um
[0466] Flow: 10 ml/min
Example 3.1
Diastereoisomer 1 of
(8S)-10-(5-chlorofuran-2-yl)-1,3,8-trimethyl-5-phenyl-7,8-dihydro-1H-pyri-
mido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[0467] SFC retention time=2.45 min
[0468] 1H NMR (400 MHz, CD.sub.2Cl.sub.2) .delta. 7.53-7.49 (3H,
m); 7.44-7.41 (2H, m); 6.13 (1H, d); 5.93 (1H, dd); 5.76 (1H, s
with splitting); 4.19 (1H, dd); 3.74 (1H, dd); 3.54 (3H, s);
3.37-3.29 (1H, m); 3.28 (3H, s); 1.17 (3H, d).
[0469] LC-MS Rt 1.31 mins; MS m/z 442 [M+H].sup.+ (Method
2minLowpHv01)
[0470] >99% de
Example 3.2
Diastereoisomer 2 of
(8S)-10-(5-chlorofuran-2-yl)-1,3,8-trimethyl-5-phenyl-7,8-dihydro-1H-pyri-
mido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[0471] SFC retention time 4.35 min
Example 3.3
(8S)-1,3,8-trimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydro-1H-pyr-
imido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00049##
[0473] The diastereoisomeric mixture was prepared analogously to
Example 3.0 from
(S)-6-(2-mercaptopropyl)-1,3-dimethyl-5-phenyl-1H-pyrrolo[3,4-d]-
pyrimidine-2,4(3H,6H)-dione (Intermediate E) and the appropriate
aldehyde.
[0474] LCMS: Rt 1.19/1.22 mins; MS m/z 439 [M+H]+; Method
2minLowpHv01
[0475] The diastereoisomers were separated under the following
conditions to afford the title compound;
[0476] Sample 110 mg in 3 ml EtOH+1.5 ml THF
[0477] Column: Chiralpak IB, 250.times.10 mm, 5 um
[0478] Mobile phase: 35% Methanol/65% CO.sub.2
[0479] Flow: 10 ml/min
Example 3.3a
Diastereoisomer 1 of
(8S)-1,3,8-trimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydro-1H-py-
rimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[0480] >99% de
[0481] SFC Retention Time: Rt 3.71
[0482] LC-MS: Rt 1.18 mins; MS 439 m/z [M+H] Method
2minLowpHv01
[0483] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.53-7.41 (5H, m); 6.84
(1H, s); 6.02 (1H, s); 4.22 (1H, dd) 3.79 (1H, dd); 3.60 (3H, s);
3.51-3.42 (1H, m); 3.35 (3H, s); 2.45 (3H, s); 1.17 (3H, d).
Example 3.4
3-(10-(5-Chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-hexahydro-1-
H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)benzonitrile
##STR00050##
[0485] A suspension comprising
3-(6-(2-mercaptoethyl)-1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrro-
lo[3,4-d]pyrimidin-5-yl)benzonitrile (Intermediate G) (150 mg,
0.441 mmol), 5-chlorofuran-2-carbaldehyde (57.5 mg, 0.441 mmol) and
bismuth triflate (28.9 mg, 0.044 mmol) in toluene (5 ml) was heated
to 100.degree. C. for 10 min using microwave radiation. After
cooling to RT, the reaction mixture was diluted with EtOAc (25 ml)
and washed with water. The layers were separated and the aqueous
extracted with EtOAc (3.times.20 ml). The combined organics were
dried over MgSO.sub.4, filtered and concentrated to yield a brown
oil. Purification was carried out by chromatography on silica
eluting with 0-45% EtOAc in iso-hexane. The relevant fraction were
combined and concentrated in vacuo to yield a pale yellow oil.
Diethyl ether (5 ml) was added to the oil to give a white solid.
The solid was isolated by suction filtration to afford the title
compound.
[0486] LCMS: Rt 1.22 in, [M+H]+ 453.4. Method 2minLowpHv01.
[0487] 1H NMR: (400 MHz, CDCl.sub.3) .delta. 7.77 (1H, d), 7.72
(2H, m), 7.64 (1H, t), 6.13 (1H, d), 5.98 (1H, dd), 5.74 (1H, s),
4.19 (1H, m), 4.11 (1H, m), 3.60 (3H, s), 3.36 (3H, s), 3.13 (1H,
m), 2.87 (1H, m).
[0488] Chiral separation of the racemate by Supercritical Fluid
Chromatography was carried out using the following conditions to
afford the compounds listed hereinafter:
[0489] Mobile Phase: 35% MeOH/65% CO.sub.2
[0490] Column: Chirapak AD-H, 250.times.10 mm, 5 um
[0491] Detection: UV @ 220 nm
[0492] Flow rate: 10 mL/min
Examples 3.5 and 3.6 are Enantiomers
Example 3.5
Enantiomer 1 of
3-(10-(5-chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-hexahydro--
1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)benzonitrile
[0493] SFC retention time=4.12 min.
[0494] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.78 (1H, d), 7.72 (2H,
mult), 7.64 (1H, t), 6.13 (1H, d), 5.98 (1H, d), 5.73 (1H, s),
4.23-4.06 (2H, mult), 3.60 (3H, s), 3.36 (3H, s), 3.13 (1H, mult),
2.87 (1H, mult).
Example 3.6
Enantiomer 2 of
3-(10-(5-chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-hexahydro--
1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)benzonitrile
[0495] SFC retention time=4.88 min.
[0496] 1H NMR: (400 MHz, CDCl.sub.3) .delta. 7.78 (1H, d), 7.72
(2H, mult), 7.64 (1H, t), 6.13 (1H, d), 5.98 (1H, d), 5.74 (1H, s),
4.24-4.07 (2H, mult), 3.60 (3H, s), 3.36 (3H, s), 3.13 (1H, mult),
2.87 (1H, mult).
[0497] The compounds of the following tabulated examples (Table 3)
were prepared analogously to Example 3.4 by replacing Intermediate
G with the appropriate starting compound (prepared by a similar
Method to Intermediate G from Intermediate F and the appropriate
halo compound in step 1) and the commercially available
aldehyde:
TABLE-US-00012 TABLE 3 Ex Structure Name LCMS NMR 3.7 ##STR00051##
10-(5-Chlorofuran-2-yl)- 1,3-dimethyl-5-(4- methylthiazol-2-
yl)-7,8-dihydro-1H- pyrimido[4',5':3,4] pyrrolo[2,1-
c][1,4]thiazine- 2,4(3H,10H)-dione LC-MS: Rt 1.26 mins; 449.4 [M +
H]+; 2 minLowpHv01; 1H NMR: (400 MHz, CDCl3) .delta. 7.22 (1H, s),
7.61 (2H, d), 5.73 (1H, s), 4.79 (1H, m), 4.54 (1H, m), 3.60 (3H,
s), 3.41 (3H, s), 3.23 (1H, m), 2.94 (1H, m), 2.60 (3H, s). 3.8
##STR00052## 10-(5-Chlorofuran-2-yl)- 5-(cyclohex-1-en-
1-yl)-1,3-dimethyl-7,8- dihydro-1H- pyrimido[4',5':3,4]
pyrrolo[2,1- c][1,4]thiazine- 2,4(3H,10H)-dione LC-MS: Rt 1.35
mins; 432.5/434.5 [M + H].sup.+; 2 minLowpHv01 1H NMR: (400 MHz,
CDCl3) .delta. 6.06 (1H, d), 5.86-5.78 (2H, m), 5.64 (1H, s), 4.30
(1H, m), 4.15 (1H, m), 3.52 (3H, s), 3.38 (3H, s), 3.10 (1H, m),
2.85 (1H, dt), 2.74-2.07 (4H, m), 1.91-1.79 (2H, m), 1.78-1.66 (2H,
m). 3.9 ##STR00053## 10-(5-Chlorofuran-2-yl)-
5-(3,5-difluorophenyl)- 1,3-dimethyl-7,8- dihydro-1H-
pyrimido[4',5':3,4] pyrrolo[2,1- c][1,4]thiazine- 2,4(3H,10H)-dione
LC-MS: Rt 1.33 mins; 464.5/466.5 [M + H].sup.+; 2 minLowpHv01 1H
NMR: (400 MHz, CDCl3) .delta. 7.01-6.90 (3H, m), 6.11 (1H, d), 5.94
(1H, dd), 5.71 (1H, s), 4.19 (1H, m), 4.10 (1H, m), 3.58 (3H, s),
3.35 (3H, s), 3.10 (1H, ddd), 2.85 (1H, ddd).
[0498] The compounds of the following tabulated examples (Table
3.1) were prepared analogously to Example 3.4 by replacing
(Intermediate G) with the appropriate starting compound (prepared
by a similar Method to Intermediate G from Intermediate F and the
appropriate halo compound in step 1) and the commercially available
aldehyde. The resulting racemates were separated by SFC
chromatographic resolution to yield single enantiomers.
TABLE-US-00013 TABLE 3.1 Structure LCMS NMR Ex Name SFC 3.1.1
##STR00054## Enantiomer 1 of
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-
methylthiazol-2-yl)-7,8-dihydro-1H- pyrimido[4',5':3,4]pyrrolo[2,1-
c][1,4]thiazine-2,4(3H,10H)-dione LC-MS: Rt 1.23 mins 449.4 [M +
H].sup.+; 2 minLowpHv01 1H NMR: (400 MHz, CDCl3) .delta. 7.14 (1H,
s), 6.11 (1H, mult), 6.01 (1H, mult), 5.73 (1H, s), 4.66 (2H,
mult), 3.59 (3H, s), 3.41 (3H, s), 3.15 (1H, mult), 2.92 (1H,
mult), 2.54 (3H, s). SFC: Column: Chiralpak IB 250 .times. 10 mm, 5
um @ 35 deg C Mobile phase: 40% Methanol/60% CO2 Flow: 10 ml/min
Detection: UV @ 220 nm Rt: 5.98 min >99% ee 3.1.2 ##STR00055##
Enantiomer 1 of 10-(5-chlorofuran-2-yl)-5-(cyclohex-1-en-
1-yl)-1,3-dimethyl-7,8-dihydro-1H- pyrimido[4',5':3,4]pyrrolo[2,1-
c][1,4]thiazine-2,4(3H,10H)-dione LC-MS: Rt 1.35 mins; 432.5/434.5
[M + H].sup.+; 2 minLowpHv01 1H NMR: (400 MHz, CDCl3) .delta. 6.06
(1H, d), 5.86-5.78 (2H, m), 5.64 (1H, s), 4.30 (1H, m), 4.15 (1H,
m), 3.52 (3H, s), 3.38 (3H, s), 3.10 (1H, m), 2.85 (1H, dt), 2.74-
2.07 (4H, m), 1.91-1.79 (2H, m), 1.78-1.66 (2H, m). SFC: Column:
Chiralpak IB 250 .times. 10 mm, 5 um @ 35 deg C Mobile phase: 25%
Methanol/75% CO2 Flow: 10 ml/min Detection: UV @ 220 nm Rt: 5.10
min >99% ee
Example 4.1
10-(5-Chlorofuran-2-yl)-1,3-diethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',5'-
:3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00056##
[0500]
1,3-Diethyl-6-(2-mercaptoethyl)-5-phenyl-1H-pyrrolo[3,4-d]pyrimidin-
e-2,4(3H,6H)-dione (Intermediate H) (329 mg, 0.958 mmol),
5-chlorofurfual (138 mg, 1.054 mmol) and bismuth triflate (62.9 mg,
0.096 mmol) were combined in toluene (Volume: 10 mL) and the
mixture heated at 95.degree. C. for 15 mins. The reaction mixture
was cooled to room temp. and evaporated under vacuum. The residue
was partitioned between DCM and sat. NaHCO.sub.3(aq) and the phases
separated. The organic phase was passed through a hydrophobic frit
and evaporated onto silica. The silica was deposited onto a 10 g
silica cartridge and the system gradient-eluted from 10-60%
EtOAc/hexane. Product eluted at 20-30% EtOAc/hexane, fractions were
combined and evaporated. The residue was triturated with
Et.sub.2O/hexane and the precipitate collected by filtration to
yield the title compound as a white solid.
[0501] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.54-7.41 (5H, m), 6.10
(1H, d), 5.93 (1H, dd), 5.48 (1H, s), 4.39 (1H, ddd), 4.17 (2H, m),
4.02 (2H, q), 3.60 (1H, ddd), 3.08 (1H, ddd), 2.84 (1H, ddd), 1.35
(3H, t), 1.21 (3H, t).
[0502] LC-MS Rt 1.37 mins; MS 456.4 m/z [M+H](Method
2minLowpHv01)
[0503] Chiral separation of the racemate by Supercritical Fluid
Chromatography was carried out using the following conditions to
afford the compounds listed hereinafter:
[0504] Mobile Phase: 45% MeOH/55% CO.sub.2
[0505] Column: SFC Chiralcel OJ-H 250.times.10 mm, 5 .mu.m diameter
@ 35.degree. C.
[0506] Detection: UV @ 220 nm
[0507] Flow rate: 10 mL/min
Example 4.2
Enantiomer 1 of
10-(5-Chlorofuran-2-yl)-1,3-diethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',5-
':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[0508] SFC retention time=5.00 mins.
[0509] LC-MS: Rt 1.41 mins; MS 456.2 m/z [M+H] Method
2minLowpHv01
[0510] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.54-7.41 (5H, m), 6.10
(1H, d), 5.93 (1H, dd), 5.48 (1H, s), 4.39 (1H, ddd), 4.17 (2H, m),
4.02 (2H, q), 3.60 (1H, ddd), 3.08 (1H, ddd), 2.84 (1H, ddd), 1.35
(3H, t), 1.21 (3H, t).
Enantiomer 2 of
10-(5-Chlorofuran-2-yl)-1,3-diethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4',5-
':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[0511] SFC retention time=6.88 mins was also isolated.
Example 5
3-Chloro-5-(10-(5-chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-he-
xahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)benzonitrile
##STR00057##
[0513]
3-Chloro-5-(1,3-dimethyl-2,4-dioxo-6-(2-(tritylthio)ethyl)-2,3,4,6--
tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
(Intermediate GA) (146 mg, 0.237 mmol),
5-chlorofuran-2-carbaldehyde (34.0 mg, 0.260 mmol) and bismuth
(III) triflate (31.0 mg, 0.047 mmol) were suspended in toluene
(Volume: 2.5 ml). The reaction was then heated to 100.degree. C.
under microwave irradiation for 3 hr. The reaction mixture was
suspended in MeOH (50 mL) and adsorbed onto silica. The residue was
purified via ISCO SiO2 12 g Redisep Rf column eluting with 20%
EtOAc/hexane. Fractions were combined and evaporated to afford a
pale yellow gum. The gum was dissolved in Et.sub.2O (30 mL) and
iso-hexane (30 mL) was added to afford a suspension. The solvent
was evaporated to afford the title compound as a pale yellow
solid.
[0514] LC-MS Rt 1.32 mins; MS m/z 487.1/489.1/491.1 [M+H]+; (Method
2minLowpHv01)
[0515] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.74 (1H, s), 7.68 (1H,
s), 7.63 (1H, s), 6.13 (1H, d), 5.98 (1H, d), 5.72 (1H, s), 4.19
(1H, dt), 4.10 (1H, m), 3.60 (3H, s), 3.36 (3H, s), 3.13 (1H, ddd),
2.87 (1H, dt).
Example 6.1
10-(2,3-Difluorophenyl)-1,3-dimethyl-5-phenyl-7,8,9,10-tetrahydropyrimido[-
4,5-a]indolizine-2,4(1H,3H)-dione
##STR00058##
[0516] Step 1:
10-(2,3-Difluorophenyl)-1,3-dimethyl-5-phenyl-7,8-dihydropyrimido[4,5-a]i-
ndolizine-2,4(1H,3H)-dione
[0517]
1,3-Dimethyl-5-phenyl-7,8-dihydropyrimido[4,5-a]indolizine-2,4(1H,3-
H)-dione (Intermediate I) (180 mg, 0.586 mmol),
1-bromo-2,3-difluorobenzene (commercially available) (0.085 mL,
0.761 mmol) and N-cyclohexyl-N-methylcyclohexanamine (commercially
available) (0.251 mL, 1.171 mmol) were combined in DMA (2.5 mL) and
the mixture sparged with nitrogen for 30 min.
[1,1'-Bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II)
(commercially available) (38.2 mg, 0.059 mmol) was then added and
the mixture heated at 110.degree. C. for 1 hour under microwave
irradiation. After cooling to room temperature, the mixture was
heated at 110.degree. C. for a further 6 hours. Further portions of
1-bromo-2,3-difluorobenzene (0.085 mL, 0.761 mmol) and
[1,1'-Bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II)
(38.2 mg, 0.059 mmol) were added, and the mixture heated at
110.degree. C. for a further 5 h. Further
[1,1'-Bis(di-tert-butylphosphino) ferrocene]dichloropalladium(II)
(38.2 mg, 0.059 mmol) and 1-bromo-2,3-difluorobenzene (0.085 mL,
0.761 mmol) were added and the mixture heated 110.degree. C. for a
further 6 hours. Further
[1,1'-Bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II)
(38.2 mg, 0.059 mmol) was added and the mixture heated at
110.degree. C. for a further 6 h. The reaction mixture was diluted
with EtOAc and washed with water and brine. The organic phase was
dried over sodium sulphate and evaporated under reduced pressure.
Purification by chromatography on silica, eluting with 15% EtOAc in
iso-hexane afforded the title compound, which was used without
further purification.
Step 2:
10-(2,3-Difluorophenyl)-1,3-dimethyl-5-phenyl-7,8,9,10-tetrahydrop-
yrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0518]
10-(2,3-Difluorophenyl)-1,3-dimethyl-5-phenyl-7,8-dihydropyrimido[4-
,5-a]indolizine-2,4(1H,3H)-dione (step 1,) (40 mg, 0.095 mmol), 10%
wt. palladium on activated carbon (10 mg, 9.54 .mu.mol) and
ammonium formate (30.1 mg, 0.477 mmol) were suspended in EtOH (5
mL) and the mixture stirred at 50.degree. C. for 19 hours, adding
extra 10% wt. palladium on activated carbon (10 mg, 9.54 .mu.mol)
after 17 hours. The reaction mixture was cooled to RT and filtered
through a Celite.RTM. (filter material) cartridge (10 g), rinsing
the residues with MeOH. The filtrates were evaporated under reduced
pressure. The residue was partitioned between DCM and water and the
phases separated. The organic phase was passed through a
hydrophobic frit and evaporated under reduced pressure to afford
the title product as a pale yellow oil.
[0519] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.45-7.34 (m, 5H), 7.01
(m, 1H), 6.91 (m, 1H), 6.33 (t, 1H), 5.08 (dd, 1H), 4.02 (ddd, 1H),
3.70 (m, 1H), 3.24 (s, 3H), 3.21 (s, 3H), 2.22 (m, 1H), 2.05 (ddd,
1H), 1.73 (m, 2H);
[0520] LC-MS Rt 1.29 min [M+H]+ 422 (Method 2minLC_v003).
Example 6.2
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimido[-
4,5-a]indolizine-2,4(1H,3H)-dione
##STR00059##
[0521] Step 1:
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8-dihydropyrimido[4,5-a]i-
ndolizine-2,4(1H,3H)-dione
[0522]
1,3-dimethyl-5-phenyl-7,8-dihydropyrimido[4,5-a]indolizine-2,4(1H,3-
H)-dione (Intermediate I), 2-bromo-5-methylfuran (commercially
available) (390 mg, 2.421 mmol), palladium acetate (commercially
available) (36.2 mg, 0.161 mmol), tri-tert-butylphosphine
tetrafluorohydroborate (commercially available) (94 mg, 0.323 mmol)
and N-cyclohexyl-N-methylcyclohexylamine (commercially available)
(0.691 mL, 3.23 mmol) were combined in dimethyl acetamide (4 mL)
and the mixture sparged with nitrogen for 30 min. The mixture was
then heated at 120.degree. C. for 15 h under microwave irradiation.
The reaction mixture was diluted with water and extracted with
EtOAc. The combined organic phases were washed with brine, dried
over sodium sulphate and evaporated under reduced pressure.
Purification by chromatography on silica, eluting with 20% EtOAc in
iso-hexane, afforded
1,3-dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8-dihydropyrimido[4,5-a]i-
ndolizine-2,4(1H,3H)-dione as a yellow solid which was used without
further purification.
Step 2:
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8,9,10-tetrahydrop-
yrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0523]
1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8-dihydropyrimido[4-
,5-a]indolizine-2,4(1H,3H)-dione (step 1) (411 mg, 1.061 mmol), 10%
wt. palladium on activated carbon (113 mg, 0.106 mmol) and ammonium
formate (669 mg, 10.61 mmol) were suspended in EtOH (30 mL) and the
mixture stirred at 50.degree. C. for 1 h. A further portion of 10%
wt. palladium on activated carbon (113 mg, 0.106 mmol) was added
and heating continued at 50.degree. C. for a further 1 hour. The
reaction mixture was cooled to RT and filtered through a
Celite.RTM. (filter material) cartridge (10 g), rinsing the residue
with MeOH. The filtrates were evaporated under reduced pressure.
The residue was taken up in a small volume of MeOH and solids
precipitated by addition of diethyl ether. The salts were removed
by filtration and rinsing with diethyl ether. The solvent was
evaporated and the residue was purified by chromatography on
silica, eluting with 20% EtOAc in iso-hexane. Precipitation from
EtOAc/iso-hexane afforded the title compound as a white solid.
[0524] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.51-7.41 (m, 5H), 5.84
(d, 1H), 5.63 (d, 1H), 4.77 (m, 1H), 4.03 (dt, 1H), 3.72 (td, 1H),
3.48 (s, 3H), 3.35 (s, 3H), 2.39 (m, 1H), 2.29 (s, 3H), 2.08 (m,
1H), 1.96-1.76 (m, 2H);
[0525] LC-MS Rt=1.17 min [M+H]+ 390.5 (Method 2minLowpH).
[0526] Chiral separation of the racemate by Supercritical Fluid
Chromatography was carried out using the following conditions to
afford the compounds listed hereinafter:
[0527] Mobile Phase: 40% MeOH/60% CO2
[0528] Column: Chiralpak AS-H, 250.times.10 mm, 5 um
[0529] Detection: UV @ 220 nm
[0530] Flow rate: 10 mL/min
[0531] Injection volume: 200 .mu.l
Examples 6.3 and 6.4 are Enantiomers
Example 6.3
(R)-1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrim-
ido[4,5-a]indolizine-2,4(1H,3H)-dione
[0532] SFC retention time=8.35 min.
[0533] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.51-7.41 (m, 5H), 5.84
(d, 1H), 5.63 (d, 1H), 4.77 (m, 1H), 4.03 (st, 1H), 3.72 (td, 1H),
3.48 (s, 3H), 3.35 (s, 3H), 2.39 (m, 1H), 2.29 (s, 3H), 2.08 (m,
1H), 1.96-1.76 (m, 2H);
[0534] LC-MS Rt=1.17 min MS m/z 390.5 [M+H](Method 2minLowpH).
[0535] Chiral purity 99% ee
Example 6.4
(S)-1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrim-
ido[4,5-a]indolizine-2,4(1H,3H)-dione
[0536] SFC retention time=7.12 min.
[0537] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.51-7.41 (m, 5H), 5.84
(d, 1H), 5.63 (d, 1H), 4.77 (m, 1H), 4.03 (st, 1H), 3.72 (td, 1H),
3.48 (s, 3H), 3.35 (s, 3H), 2.39 (m, 1H), 2.29 (s, 3H), 2.08 (m,
1H), 1.96-1.76 (m, 2H);
[0538] LC-MS Rt=1.17 min MS m/z 390.5 [M+H](Method 2minLowpH).
[0539] Chiral purity >99% ee
X-ray Crystallography
[0540] The absolute stereochemistry of the enantiomers was
confirmed by X-ray crystallography of
(S)-1,3-Dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8,9,10-tetrahydropyri-
mido[4,5-a]indolizine-2,4(1H,3H)-dione.
##STR00060##
TABLE-US-00014 TABLE 1 Crystal data and structure refinement
Empirical formula C23H23N3O3 Formula weight 389.44 Temperature
100(2) K Wavelength 1.54178 .ANG. Crystal system Orthorhombic Space
group P212121 Unit cell dimensions a = 6.391(2) .ANG. .alpha. =
90.degree.. b = 7.731(2) .ANG. .beta. = 90.degree.. c = 40.480(11)
.ANG. .gamma. = 90.degree.. Volume 2000.1(10) .ANG..sup.3 Z 4
Density (calculated) 1.293 Mg/m.sup.3 Absorption coefficient 0.703
mm.sup.-1 F(000) 824 Crystal size 0.30 .times. 0.14 .times. 0.07
mm.sup.3 Theta range for data 2.18 to 68.18.degree.. collection
Index ranges -7 <= h <= 7, -9 <= k <= 9, -48 <= l
<= 48 Reflections collected 39930 Independent reflections 3651
[R(int) = 0.0412] Completeness to 100.0% theta = 68.18.degree.
Absorption correction Semi-empirical from equivalents Max. and min.
transmission 0.9525 and 0.8169 Refinement method Full-matrix
least-squares on F.sup.2 Data/restraints/parameters 3651/0/265
Goodness-of-fit on F.sup.2 1.111 Final R indices R1 = 0.0295, wR2 =
0.0753 [I > 2sigma(I)] R indices (all data) R1 = 0.0297, wR2 =
0.0754 Absolute structure 0.05(17) parameter Largest diff. peak and
hole 0.188 and -0.175 e .ANG..sup.-3
Example 6.5
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimid-
o[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00061##
[0541] Step 1:
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydropyrimido[4,5-a-
]indolizine-2,4(1H,3H)-dione
[0542] 4-Methyl-2-tributylstannanyl-thiazole (Intermediate J) (491
mg, 1.265 mmol) was added to a suspension of
trifluoro-methanesulfonic acid
2,4-dimethyl-1,3-dioxo-9-phenyl-1,2,3,4,7,8-hexahydro-2,4,8a-triaza-fluor-
en-5-yl ester (Intermediate K) (480 mg, 1.054 mmol), lithium
chloride (4.47 mg, 0.105 mmol), copper(I) iodide (20.07 mg, 0.105
mmol) and PdCl.sub.2(dppf) (77 mg, 0.105 mmol) in THF (12 ml) and
the mixture heated at reflux under nitrogen for 2 h. Further
4-methyl-2-tributylstannanyl-thiazole (Intermediate J) (491 mg,
1.265 mmol) was added and the mixture continued at reflux for 2 h,
then further 4-methyl-2-tributylstannanyl-thiazole (Intermediate J)
(491 mg, 1.265 mmol) was added and the mixture stirred at reflux
for 1 h. The mixture was cooled to RT, diluted with EtOAc and
washed with dilute aqueous NH.sub.4OH, water and brine. The organic
phase was dried over sodium sulphate and evaporated under reduced
pressure. The residue was triturated with MeOH/Et.sub.2O and the
precipitate collected by filtration. The precipitate was purified
by chromatography on silica, eluting with 50% EtOAc in hexane.
Trituration with Et.sub.2O and collection by filtration afforded
1,3-dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydropyrimido[4,5-a-
]indolizine-2,4(1H,3H)-dione as a pale yellow solid
Step 2:
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8,9,10-tetrahydr-
opyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0543]
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydropyrimido-
[4,5-a]indolizine-2,4(1H,3H)-dione (from Step 1) (174 mg, 0.430
mmol), Pd/C (commercially available) (45.8 mg, 0.043 mmol) and
ammonium formate (271 mg, 4.30 mmol) were combined in EtOH (8 ml)
and the mixture heated at 60.degree. C. for 2 days. Further
portions of Pd/C (45.8 mg, 0.043 mmol) and ammonium formate (271
mg, 4.30 mmol) were added as required to ensure completion of the
reaction. The reaction mixture was cooled to RT and filtered
through a Celite.RTM. (filter material) cartridge (10 g), rinsing
the residue with MeOH. The filtrates were evaporated under reduced
pressure. The residue was partitioned between DCM and water and the
phases separated. The organic phase was passed through a
hydrophobic frit and evaporated under vacuum. The residue was
purified by chromatography on silica, eluting from 50% EtOAc in
hexane to 80% EtOAc in hexane. Precipitation from Et.sub.2O/hexane
afforded the title compound as a white solid.
[0544] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.52-7.45 (m, 5H), 6.78
(d, 1H), 5.14 (dd, 1H), 4.10 (ddd, 1H), 3.75 (ddd, 1H), 3.42 (s,
3H), 3.36 (s, 3H), 2.59 (m, 1H), 2.48 (d, 3H), 2.32 (m, 1H), 1.88
(m, 2H).
[0545] LC-MS Rt 1.03 min [M+H].sup.+ 407.5 (Method 2minLowpH)
[0546] Chiral separation of the racemate by Supercritical Fluid
Chromatography was carried out using the following conditions to
afford the compounds listed hereinafter:
[0547] Mobile Phase: 50% IPA+0.1% DEA/50% CO.sub.2
[0548] Column: Chiralpak ID, 250.times.10 mm, 5 um
[0549] Detection: UV @ 220 nm
[0550] Flow rate: 10 mL/min
[0551] Injection volume: 200 .mu.l
Examples 6.6 and 6.7 are Enantiomers
Example 6.6
(R)-1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8,9,10-tetrahydropyr-
imido[4,5-a]indolizine-2,4(1H,3H)-dione
[0552] SFC retention time=8.21 min.
[0553] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.52-7.45 (m, 5H), 6.78
(d, 1H), 5.14 (dd, 1H), 4.10 (ddd, 1H), 3.75 (ddd, 1H), 3.42 (s,
3H), 3.36 (s, 3H), 2.59 (m, 1H), 2.48 (d, 3H), 2.32 (m, 1H), 1.88
(m, 2H);
[0554] LC-MS Rt=1.03 min MS m/z 407.5 [M+H](Method 2minLowpH).
[0555] Chiral purity >99% ee
Example 6.7
(S)-1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8,9,10-tetrahydropyr-
imido[4,5-a]indolizine-2,4(1H,3H)-dione
[0556] SFC retention time=4.75 min was also isolated.
[0557] Chiral purity >99% ee
Example 7.1
5-(3-Fluorophenyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetrahydr-
o pyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00062##
[0559] To a stirred suspension of 2-methylfuran (31.1 mg, 0.379
mmol) and
5-(3-fluorophenyl)-10-hydroxy-1,3-dimethyl-7,8,9,10-tetrahydropyrimido[4,-
5-a]indolizine-2,4(1H,3H)-dione (Intermediate L) (100 mg, 0.291
mmol) in MeCN (2912 .mu.L) was added gold (III) chloride (8.83 mg,
0.029 mmol) at ambient temperature. Upon addition the white
suspension formed a dark brown solution. The reaction was left for
30mins. The reaction mixture was concentrated in-vacuo, then
diluted with DCM (10 mL) and water (10 mL). The biphasic solution
was passed through a phase-separator, then concentrated in-vacuo to
afford a brown oil. The residue was purified using the Agilent
Prep. System (50-98%, low pH) to the title compound as an off-white
solid.
[0560] LCMS Rt 1.33 mins; MS m/z 408.6 [M+H]+; Method
2minlowpHv01.
Examples 7.2 and 7.3 are Enantiomers.
[0561] Chiral separation of the racemate by Supercritical Fluid
Chromatography was carried out using the following conditions to
afford the compounds listed hereinafter:
[0562] Mobile Phase: 30% MeOH/70% CO.sub.2
[0563] Column: 2.times. Chiralpak ID coupled 250.times.10 mm, 5 um
@35 deg C.
[0564] Detection: UV @ 220 nm
[0565] Flow rate: 10 mL/min
Example 7.2
Enantiomer 1 of
5-(3-fluorophenyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetrahyd-
ropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0566] SFC retention time=14.40 mins:
[0567] 1H NMR (400 MHz, CDCl3) .delta. 7.43 (1H, q), 7.26-7.22 (1H,
m), 7.19-7.11 (2H, m), 5.85 (1H, d), 5.63 (1H, d), 4.77 (1H, t),
4.06-3.99 (1H, m), 3.77-3.69 (1H, m), 3.48 (3H, s), 3.37 (3H, s),
2.44-2.37 (1H, m), 2.30 (3H, s), 2.13-2.04 (1H, m), 1.95-1.79 (2H,
m).
[0568] LCMS Rt 1.30 mins; MS m/z 408.4 [M+H]+;
(Method)2minlowpHV01.
[0569] >99% ee
Example 7.3
Enantiomer 2 of
5-(3-fluorophenyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetrahyd-
ropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0570] SFC retention time=13.07 mins was also isolated:
[0571] >99% ee
[0572] The following listed examples were prepared in a similar
manner to Example 7, replacing
5-(3-fluorophenyl)-10-hydroxy-1,3-dimethyl-7,8,9,10-tetrahydropyrimido[4,-
5-a]indolizine-2,4(1H,3H)-dione (Intermediate L) with the
appropriate starting material (either Intermediate La or
Intermediate Lb). The mixtures of diastereomers were resolved by
SFC chromatographic resolution under the listed conditions to
afford the title compounds.
Example 7.4
(9R,10R)-5-(3-fluorophenyl)-1,3,9-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,-
10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione or
(9R,10S)-5-(3-fluorophenyl)-1,3,9-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,-
10-tetrahydro pyrimido[4,5-a]indolizine-2,4(1H,3H)-dione or
(9S,10R)-5-(3-fluorophenyl)-1,3,9-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,-
10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione or
(9S,10S)-5-(3-fluorophenyl)-1,3,9-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,-
10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00063##
[0574] Separation conditions:
[0575] Column: Chiralcel OD-H 250.times.10 mm, 5 um @ 35 deg C.
[0576] Mobile phase: 25% Isopropanol/75% CO2
[0577] Flow: 10 ml/min
[0578] Detection: UV @ 220 nm
Diastereomer 1 of
5-(3-fluorophenyl)-1,3,9-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetra-
hydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0579] Rt=4.49 mins
[0580] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.45 (1H, mult), 7.26
(1H, mult), 7.20-7.12 (2H, mult), 5.85 (1H, d), 5.61 (1H, d), 4.38
(1H, mult), 3.96-3.81 (2H, mult), 3.45 (3H, s), 3.35 (3H, s), 2.54
(1H, mult), 2.29 (3H, s), 2.06 (1H, mult), 1.64-1.58 (1H, mult),
1.26 (3H, d).
[0581] LC-MS Rt 1.34 mins [M+H].sup.+ 422.2 (Method
2minLowpHv01).
Example 7.5a and 7.5b
(8R,10R)-5-(3-Fluorophenyl)-1,3,8-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,-
10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione or
(8S,10R)-5-(3-fluorophenyl)-1,3,8-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,-
10-tetrahydro pyrimido[4,5-a]indolizine-2,4(1H,3H)-dione or
(8R,10S)-5-(3-fluorophenyl)-1,3,8-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,-
10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione or
(8S,10S)-5-(3-fluorophenyl)-1,3,8-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,-
10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00064##
[0583] Separation conditions:
[0584] Column: 2 coupled Chiralcel OD-H 250.times.10 mm, 5 um @ 35
deg C.
[0585] Mobile phase: 15% Methanol/85% CO2
[0586] Flow: 10 ml/min
[0587] Detection: UV @ 220 nm
Example 7.5a
Diastereomer 1 of
5-(3-fluorophenyl)-1,3,8-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetra-
hydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0588] Rt=11.80 mins
[0589] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.45 (1H, m), 7.23 (1H,
d), 7.15 (2H, m), 5.83 (1H, dd), 5.58 (1H, dd), 4.76 (1H, d), 3.95
(1H, dd), 3.46 (3H, s), 3.35 (3H, s), 3.27 (1H, br t), 2.34 (1H,
d), 2.29 (3H, s), 2.06 (1H, m), 1.79 (1H, td), 1.00 (3H, d).
[0590] LC-MS Rt 1.35 mins [M+H].sup.+422.3 (Method
2minLowpHv01)
Example 7.5b
Diastereomer 2 of
5-(3-fluorophenyl)-1,3,8-trimethyl-10-(5-methylfuran-2-yl)-7,8,9,10-tetra-
hydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0591] Rt=13.82 mins
[0592] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.45 (1H, m), 7.20 (1H,
d), 7.15 (2H, m), 5.87 (1H, dd), 5.72 (1H, dd), 4.68 (1H, t), 3.83
(1H, dq), 3.47 (1H, br t), 3.35 (3H, s), 3.33 (3H, s), 2.52 (1H,
m), 2.29 (3H, s), 2.01 (1H, m), 1.68 (1H, m), 1.03 (3H, d)
[0593] LC-MS Rt 1.34 mins [M+H].sup.+ 422.4 (Method
2minLowpHv01)
Example 7.6
(9R,10R)-5-(3-Fluorophenyl)-1,3,9-trimethyl-10-(4-methylthiazol-2-yl)-7,8,-
9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione or
(9S,10R)-5-(3-fluorophenyl)-1,3,9-trimethyl-10-(4-methylthiazol-2-yl)-7,8,-
9,10-tetrahydro pyrimido[4,5-a]indolizine-2,4(1H,3H)-dione or
(9R,10S)-5-(3-fluorophenyl)-1,3,9-trimethyl-10-(4-methylthiazol-2-yl)-7,8,-
9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione or
(9S,10S)-5-(3-fluorophenyl)-1,3,9-trimethyl-10-(4-methylthiazol-2-yl)-7,8,-
9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00065##
[0595] Separation conditions:
[0596] Column: Chiralpak AD 250.times.10 mm, 5 um @ 35.2.degree.
C.
[0597] Mobile phase: 25% Methanol/75% CO2
[0598] Flow: 10 ml/min
[0599] Detection: UV @ 220 nm
Diastereomer 1 of
5-(3-fluorophenyl)-1,3,9-trimethyl-10-(4-methylthiazol-2-yl)-7,8,9,10-tet-
rahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0600] Rt=4.22 mins
[0601] 1H NMR (400 MHz, CDCl3) .delta. 7.47 (1H, mult), 7.26 (1H,
d), 7.21-7.14 (2H, mult), 6.82 (1H, s), 5.09 (1H, d), 4.19 (1H,
dd), 3.79 (1H, td), 3.47 (3H, s), 3.31 (3H, s), 2.49-2.40 (4H,
mult), 2.06 (1H, mult), 1.75 (1H, mult), 1.09 (3H, d).
[0602] LC-MS Rt 1.22 mins [M+H].sup.+ 439.6 (Method
2minLowpHv01)
Example 8.1
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-m-tolyl-7,8,9,10-tetrahydropyrimi-
do[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00066##
[0604]
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-m-tolyl-7,8-dihydropyrimid-
o[4,5-a]indolizine-2,4(1H,3H)-dione (Intermediate M) (100 mg, 0.24
mmol) was dissolved in ethyl acetate (20 mL) and the resulting
solution was hydrogenated using the H-Cube.RTM. (10% platinum on
carbon CatCart.RTM.), at atmospheric pressure of hydrogen and
20.degree. C., for 6.5 hours. The solvent was removed under vacuum.
The crude material was purified by chromatography on silica,
eluting with 1-2% MeOH/DCM to obtain a gum. Diethyl ether was added
and removed under vacuum to give the title compound as a solid.
[0605] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.39 (1H, t), 7.30-7.23
(3H, mult), 6.88 (1H, s), 5.33 (1H, mult), 4.07 (1H, mult), 3.72
(1H, mult), 3.40 (3H, s), 3.34 (3H, s), 2.69 (1H, mult), 2.56 (3H,
s), 2.45-2.36 (4H, mult), 1.92-1.82 (2H, mult);
[0606] LC-MS Rt=1.20 min [M+H].sup.+ 421.5 (Method
2minLowpHv01).
[0607] Chiral separation of the racemate by Supercritical Fluid
Chromatography was carried out using the following conditions to
afford the compounds listed hereinafter:
[0608] Mobile Phase: 25% IPA/75% CO.sub.2
[0609] Column: Chiralpak IB 250.times.10 mm, 5 .mu.M
[0610] Detection: UV @ 220 nM
[0611] Flow rate: 10 mL/min
[0612] Injection volume: 50 .mu.L
Example 8.2
Enantiomer 1 of
1,3-dimethyl-10-(4-methylthiazol-2-yl)-5-m-tolyl-7,8,9,10-tetrahydropyrim-
ido[4,5-a]indolizine-2,4(1H,3H)-dione
[0613] First eluted peak Rt=5.15 min
[0614] .sup.1H NMR (400 MHz, CDCl3) .delta. 7.38 (1H, t), 7.27-7.22
(3H, mult), 6.76 (1H, s), 5.10 (1H, dd), 4.06 (1H, mult), 3.71 (1H,
mult), 3.41 (3H, s), 3.35 (3H, s), 2.56 (1H, mult), 2.47 (3H, s),
2.43 (3H, s), 2.30 (1H, mult), 1.96-1.80 (2H, mult);
[0615] LC-MS Rt=1.19 min [M+H].sup.+ 421.2 (Method
2minLowpHv01);
[0616] Chiral purity >99.9% ee.
Example 8.3
Enantiomer 1 of
5-(3-Chlorophenyl)-10-(4-chlorothiazol-2-yl)-1,3-dimethyl-7,8,9,10-tetrah-
ydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00067##
[0618] The title compound was prepared analogously to Example 8.1.
The racemate was separated by SFC chromatographic resolution under
the following conditions:
[0619] Chiralpak AD 250.times.10 mm
[0620] Co-Solvent IPA 35%
[0621] Total Flow 10 ml/min
[0622] Column Temp. 34.9.degree. C.
[0623] SFC Retention time=5.03 min
[0624] LC-MS Rt 1.28 mins; m/z 461.4 [M+H].sup.+; Method
2minlowpHV01.
[0625] 1H NMR (400 MHz, CDCl3) .delta. 7.47-7.41 (3H, m), 7.40-7.34
(1H, m), 7.02 (1H, s), 5.13 (1H, t), 4.11-4.04 (1H, m), 3.80-3.73
(1H, m), 3.41 (3H, s), 3.36 (3H, s), 2.68-2.61 (1H, m), 2.35-2.26
(1H, m), 1.95-1.80 (2H, m).
Example 9.0
3-(1,3-Dimethyl-10-(4-methylthiazol-2-yl)-2,4-dioxo-1,2,3,4,7,8,9,10-octah-
ydro pyrimido[4,5-a]indolizin-5-yl)benzonitrile
##STR00068##
[0626] Step 1:
1,3-Dimethyl-6-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,4-d]pyrimi-
dine-2,4(3H,6H)-dione
[0627] Sodium hydride (60% in mineral oil, 335 mg, 8.4 mmol) was
added to an ice cooled partial suspension of
1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
(Intermediate F step 2) (1.00 g, 5.6 mmol), SEM-Cl (1.485 mL, 8.4
mmol) and benzyl triethylammonium chloride (76 mg, 0.34 mmol) in
THF (15 mL). The mixture was allowed to reach room temperature
slowly and was stirred for 18 hours. The reaction mixture was
quenched cautiously with sat ammonium chloride solution (80 mL,
added dropwise), then was extracted with ethyl acetate (3.times.40
mL). The combined organic layers were washed with water (1.times.40
mL), brine (1.times.40 mL) then dried with magnesium sulfate and
the solvent was removed under vacuum. The residue was triturated
with iso-hexane and was dried under vacuum at 50.degree. C.
[0628] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.38 (1H, s), 6.47 (1H,
s), 5.26 (2H, s), 3.48 (2H, t), 3.41 (3H, s), 3.40 (3H, s), 0.93
(2H, t), 0.00 (9H, s).
[0629] LC-MS Rt 1.15 mins; MS m/z 310.4 [M+H]+; (Method
2minLowpHv01)
Step 2:
1,3-Dimethyl-2,4-dioxo-6-((2-(trimethylsilyl)ethoxy)methyl)-2,3,4,-
6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-ylboronic acid
[0630] Butyl lithium (1.26M, 28.4 mL, 35.8 mmol) was added dropwise
to a solution of diisopropylamine (3.63 g, 35.8 mmol) in THF (20
mL) at -78.degree. C., keeping the internal temperature below
-40.degree. C. Once the addition was complete, the contents of the
flask were allowed to warm to -5.degree. C., then were re-cooled to
-78.degree. C. The resulting mixture was cannulated over a period
of about 30 minutes into a suspension of
1,3-dimethyl-6-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,4-d]pyrimi-
dine-2,4(3H,6H)-dione (step 1) (6.93 g, 22.4 mmol) in THF (75 mL)
at -78.degree. C. The mixture was stirred at -78.degree. C. for 30
minutes, then triisopropylborate (8.3 mL, 35.8 mmol) was added
dropwise. The solution was stirred at -78.degree. C. for 1.5 hours,
then was quenched by carefully adding sat ammonium chloride (200
mL).
[0631] The mixture was allowed to reach room temperature and was
extracted with ethyl acetate (3.times.100 mL). The combined organic
extracts were dried with magnesium sulfate and the solvent was
removed under vacuum. The crude material was triturated with
ether/hexane and dried under vacuum at 50.degree. C. The solid was
recombined with the mother liquor which was reduced under vacuum.
The resultant semi-solid material was triturated with iso-hexane
and dried under vacuum at 50.degree. C. to afford the title
compound;
[0632] LC-MS: Rt 1.19 mins; MS m/z 354.4 [M+H]+; (Method
2minLowpHv01)
Step 3:
3-(1,3-Dimethyl-2,4-dioxo-6-((2-(trimethylsilyl)ethoxy)methyl)-2,3-
,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
[0633] A mixture of
1,3-dimethyl-2,4-dioxo-6-((2-(trimethylsilyl)ethoxy)methyl)-2,3,4,6-tetra-
hydro-1H-pyrrolo[3,4-d]pyrimidin-5-ylboronic acid (step 2) (2.0 g,
5.7 mmol), 3-bromobenzonitrile (937 mg, 5.2 mmol), Pd-118 (168 mg,
0.26 mmol) and potassium carbonate (1.42 g, 10.3 mmol) in n-butyl
acetate (40 mL) was heated to 80.degree. C., then water (2.23 mL,
124.0 mmol) was added and the mixture was heated at 80.degree. C.
for 90 minutes. The reaction mixture was diluted with water (100
mL), the layers were separated as much as possible, and the aqueous
phase (and residual organic portion) was extracted with DCM
(1.times.100 mL, 2.times.50 mL). The combined organic extracts were
washed with brine (1.times.100 mL), dried with magnesium sulfate
and the solvent was removed under vacuum.
[0634] The resulting residue was re-crystallised from methanol and
dried under vacuum at 50.degree. C. to afford the title
compound;
[0635] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.87-7.82 (2H, mult),
7.74 (1H, d), 7.59 (1H, t), 6.59 (1H, s), 5.14 (2H, s), 3.50 (2H,
t), 3.43 (3H, s), 3.56 (3H, s), 0.91 (2H, t), 0.00 (9H, s).
[0636] LC-MS: Rt 1.34 mins; MS m/z 411.4 [M+H]+; (Method
2minLowpHv01)
Step 4:
3-(1,3-Dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyri-
midin-5-yl)benzonitrile
[0637] TBAF solution (20.3 mL, 20.3 mmol) was added to a suspension
of
3-(1,3-dimethyl-2,4-dioxo-6-((2-(trimethylsilyl)ethoxy)methyl)-2,3,4,6-te-
trahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile (step 3) (832
mg, 2.0 mmol) in THF (6 mL), giving a solution which was stirred at
60.degree. C. for 1 hour. Most of the organic solvent was removed
from the reaction mixture, water (100 mL) was added and the mixture
was stirred for 5 minutes The aqueous suspension was extracted with
choroform (4.times.100 mL). A large amount of the mixture remained
as an emulsion. Sat brine (100 mL) was added to break up the
emulsion, and the aqueous phase was extracted with more chloroform
(4.times.100 mL). The combined organic extracts were dried with
magnesium sulfate and the solvent was removed under vacuum to yield
a red oil. The crude red oil was triturated with methanol to give a
pink solid which was dried under vacuum at 50.degree. C. The solid
was triturated with methanol again and dried under vacuum at
50.degree. C.
[0638] LC-MS: Rt 0.91 mins; MS m/z 281.4 [M+H]+; (Method
2minLowpHv1)
Step 5: Methyl
4-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)butanoate
[0639] A mixture of
3-(1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-
-yl)benzonitrile (step 4) (300 mg, 1.07 mmol), methyl
4-bromobutanoate (291 mg, 1.61 mmol) and cesium carbonate (697 mg,
2.14 mmol) in DMF (4 mL) was stirred at 80.degree. C. for 2 hours.
The reaction mixture was diluted with ethyl acetate (20 mL) and was
washed with 1M HCl (3.times.10 mL). After the third wash, the
mixture was concentrated in vacuo. The residue was triturated with
diethyl ether and dried under vacuum at 50.degree. C.
[0640] LC-MS: Rt 0.98 mins; MS m/z 381.5 [M+H]+; (Method
2minLowpHv1)
Step 6:
4-(5-(3-Cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo-
[3,4-d]pyrimidin-6(2H)-yl)butanoic acid
[0641] Lithium hydroxide (965 mg, 23.0 mmol) was added to a
suspension of methyl
4-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo-
[3,4-d]pyrimidin-6(2H)-yl)butanoate (Step 5) (1.75 g, 4.6 mmol) in
THF (9 mL)/water (9 mL). The mixture was stirred at room
temperature for 4 hours. The organic solvent was largely removed
under vacuum, the residue was acidified to pH 1, using 2M HCl. The
resulting slurry was diluted with water (100 mL). The mixture was
reduced under vacuum to give a white solid, which was further dried
under vacuum at 50.degree. C. overnight.
[0642] 1H NMR (400 MHz, DMSO-d6) .delta. 12.14 (1H, v br),
7.95-7.89 (2H, mult), 7.78 (1H, d), 7.67 (1H, t), 7.05 (1H, s),
3.94 (2H, t), 3.16 (3H, s), 2.09 (2H, t), 1.82 (2H, t).
[0643] LC-MS: Rt 0.88 mins; MS m/z 367.2 [M+H]+; (Method
2minLowpHv02)
Step 7:
3-(1,3-Dimethyl-2,4,10-trioxo-1,2,3,4,7,8,9,10-octahydropyrimido[4-
,5-a]indolizin-5-yl)benzonitrile
##STR00069##
[0645] T3P.RTM. solution (2.45 mL, 4.2 mmol) was added to a
solution of
4-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)butanoic acid (step 6) (1.54 g, assume 4.2 mmol)
in DMF (9 mL) and the mixture was stirred at 100.degree. C. for 5
hours. A further 2.45 mL (4.2 mmol) of T3P.RTM. solution was added
after 3 hours and a further portion (1.24 mL, 2.1 mmol) was added
after 4 hours. The reaction mixture was diluted with ethyl acetate
(100 mL) and washed with sat. sodium bicarbonate (2.times.50 mL)
and brine (2.times.50 mL). The organic phase was dried with
magnesium sulfate and the solvent was removed under vacuum.
[0646] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.83 (1H, d), 7.78-7.72
(2H, mult), 7.67 (1H, t), 4.01 (2H, mult), 3.92 (3H, s), 3.36 (3H,
s), 2.74 (2H, t), 2.26 (2H, mult).
[0647] LC-MS: Rt 0.94 mins; MS m/z 349.2 [M+H]+; (Method
2minLowpHv1)
Step 8:
5-(3-Cyanophenyl)-1,3-dimethyl-2,4-dioxo-1,2,3,4,7,8-hexahydropyri-
mido[4,5-a]indolizin-10-yl trifluoromethanesulfonate
##STR00070##
[0649] Trifluoromethanesulfonic anhydride (221 .mu.L, 1.31 mmol)
was added to an ice-cooled solution of
3-(1,3-dimethyl-2,4,10-trioxo-1,2,3,4,7,8,9,10-octahydropyrimido[4,5-a]in-
dolizin-5-yl)benzonitrile (step 7) (350 mg, 1.01 mmol) and
2,6-lutidine (176 .mu.L, 1.51 mmol) in DCM (5 mL). The solution was
stirred at ice-bath temperature for 2 hours, further portions of
2,6-lutidine (88 .mu.L, 0.78 mmol) and trifluoromethanesulfonic
anhydride (85 .mu.L, 0.50 mmol) were added. The reaction mixture
was diluted with dichloromethane (40 mL) and was washed with water
(2.times.20 mL) and sat. brine (1.times.20 mL). The organic phase
was dried with magnesium sulfate and the solvent was removed under
vacuum.
[0650] LC-MS: Rt 1.20 mins; MS m/z 481.5 [M+H]+; (Method
2minLowpHv01)
Step 9:
3-(1,3-Dimethyl-2,4-dioxo-10-(4,4,5,5-tetramethyl-1,3,2-dioxaborol-
an-2-yl)-1,2,3,4,7,8-hexahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
##STR00071##
[0652] A mixture of
5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-1,2,3,4,7,8-hexahydropyrimido[4,-
5-a]indolizin-10-yl trifluoromethanesulfonate (step 8) (480 mg, 1.0
mmol), bis(pinaolato)diboron (279 mg, 1.1 mmol),
bis(triphenylphosphine)palladium(II) chloride (21 mg, 0.03 mmol),
triphenylphosphine (16 mg, 0.06 mol) and potassium phenoxide (J.
Am. Chem. Soc., 1959, Vol. 81, pp 2705-2715, 198 mg, 1.5 mmol) in
toluene (10 mL) was stirred at 60.degree. C., under nitrogen, for 3
hours. The reaction mixture was diluted with water (20 mL) and
extracted with ethyl acetate (3.times.20 mL). The combined organic
extracts were washed with water (2.times.20 mL), brine (1.times.20
mL), dried with magnesium sulfate and the solvent was removed under
vacuum. Purified by chromatography on silica, eluting with 20-50%
EtOAc/hexane. The combined fractions were concentrated in vacuo to
yield title product.
[0653] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.78 (1H, d), 7.75-7.71
(2H, mult), 7.60 (1H, t), 6.82 (1H, t), 3.81 (2H, t), 3.59 (3H, s),
3.37 (3H, s), 2.43 (2H, mult), 1.36 (12H, s).
[0654] LC-MS: Rt 1.25 mins; MS m/z 459.6 [M+H]+; (Method
2minLowpHv01)
Step 10:
3-(1,3-Dimethyl-10-(4-methylthiazol-2-yl)-2,4-dioxo-1,2,3,4,7,8-h-
exahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
##STR00072##
[0656] A mixture of
3-(1,3-dimethyl-2,4-dioxo-10-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl-
)-1,2,3,4,7,8-hexahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
(step 9) (202 mg, 0.44 mmol), 2-iodo-4-methylthiazole (Intermediate
O) (90 mg, 0.40 mmol), dichloro[1,1'
bis(di-tert-butylphosphino)]ferrocene palladium (11) (13 mg, 0.02
mmol) and barium hydroxide (137 mg, 0.80 mmol) in
acetonitrile/water (1:1, 2 mL) was stirred at room temperature for
45 minutes and at 80.degree. C. for 1 hour. The reaction mixture
was diluted with 1M HCl (10 mL) and was extracted with DCM
(3.times.10 mL). The combined organic layers were filtered through
a hydrophobic frit and the solvent removed under vacuum. Purified
by chromatography on silica, eluting with 0.5% MeOH/DCM, 1%
MeOH/DCM, 1.5-2.0% MeOH afforded the title product:
[0657] 1H NMR (400 MHz, CDCl.sub.3) d 7.82-7.74 (3H, mult), 7.62
(1H, t), 6.93 (1H, s), 6.52 (1H, t), 3.91 (2H, t), 3.34 (3H, s),
2.81 (3H, s), 2.60 (2H, mult), 2.50 (3H, s).
[0658] LC-MS: Rt 1.10 mins; MS m/z 430.1 [M+H]+; (Method
2minLowpHv01)
Step 11:
3-(1,3-Dimethyl-10-(4-methylthiazol-2-yl)-2,4-dioxo-1,2,3,4,7,8,9-
,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
##STR00073##
[0660] A solution of
3-(1,3-dimethyl-10-(4-methylthiazol-2-yl)-2,4-dioxo-1,2,3,4,7,8-hexahydro-
pyrimido[4,5-a]indolizin-5-yl)benzonitrile (step 10) (76 mg, 0.18
mmol) in ethanol/THF (1:1, 30 mL) was hydrogenated over 10%
platinum on carbon, at room temperature and atmospheric pressure of
hydrogen for 19 hours. A further portion of platinum on carbon
catalyst was added (10 mg) and hydrogenation was continued as
before for 24 hours. A further portion of platinum on carbon
catalyst was added (60 mg) and hydrogenation was continued as
before for a further 18 hours. The reaction mixture was filtered
through GF/F paper to remove the catalyst, washing well with 20%
MeOH/DCM. The filtrate was reduced under vacuum. The crude material
was re-dissolved in EtOH/THF (1:1, 20 mL), the solution was
de-gassed and 10% platinum on carbon (60 mg) was added. The mixture
was hydrogenated at room temperature and atmospheric pressure of
hydrogen for 20 hours. The reaction mixture was filtered through
GF/F paper to remove the catalyst, washing well with 20% MeOH/DCM.
The filtrate was reduced under vacuum. Purified by chromatography
on silica, eluting with 0.5-1.5% MeOH/DCM afforded a yellow oily
residue. Diethyl ether was added to the residue and removed under
vacuum to give a white solid which was dried under vacuum at
50.degree. C.
[0661] 1H NMR (400 MHz, CDCl.sub.3) 7.81-7.70 (3H, mult), 7.62 (1H,
mult), 6.81 (1H, s), 5.18 (1H, s), 4.06 (1H, d), 3.78 (1H, mult),
3.42 (3H, s), 3.36 (3H, s), 2.61 (1H, d), 2.49 (3H, s), 2.34 (1H,
mult), 1.93 (2H, mult).
[0662] LC-MS: Rt 1.05 mins; MS m/z 432.1 [M+H].sup.+; (Method
2minLowpHv02)
[0663]
3-(1,3-Dimethyl-10-(4-methylthiazol-2-yl)-2,4-dioxo-1,2,3,4,7,8,9,1-
0-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile was purified
by chiral separation under these conditions to afford the compound
listed hereinafter:
[0664] Column: Chiralpak IB 250.times.10 mm, 5 um @35 deg C.
[0665] Mobile Phase: 35% MeOH/65% CO2
[0666] Flow: 10 ml/min
[0667] Detection: UV@220 um
Example 9.1
Enantiomer 1 of
3-(1,3-dimethyl-10-(4-methylthiazol-2-yl)-2,4-dioxo-1,2,3,4,7,8,9,10-octa-
hydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
[0668] SFC Retention Time 4.09 mins
[0669] 1H NMR (400 MHz, CDCl.sub.3) 7.82-7.73 (3H, m), 7.63 (1H,
m), 6.79 (1H, s), 5.12 (1H, s), 4.05 (1H, m), 3.78 (1H, m), 3.42
(3H, s), 3.35 (3H, s), 2.59 (1H, m), 2.47 (3H, s), 2.32 (1H, m),
2.00-1.86 (2H, m).
[0670] LC-MS: Rt 1.08 mins; MS m/z 432.2 [M+H].sup.+; (Method
2minLowpHv01)
[0671] 100% ee
Example 9.2
3-(1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4,7,8,9,10-octahydropyrimido[4,5--
a]indolizin-10-yl)benzonitrile
##STR00074##
[0673] The title compound was prepared analogously to Example 9 by
replacing 2-iodo-4-methylthiazole (step 10) with
3-bromobenzonitrile;
[0674] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.58-7.44 (7H, mult),
7.32-7.24 (2H, mult), 4.91 (1H, mult), 4.10 (1H, mult), 3.80 (1H,
mult), 3.35 (3H, s), 3.27 (3H, s), 2.37 (1H, mult), 2.10 (1H,
mult), 1.85-1.70 (2H, mult).
[0675] LC-MS Rt 1.19 mins [M+H].sup.+ 411.5 (Method
2minLowpHv01)
Example 10.0
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimid-
o[4',5':3,4]pyrrolo[1,2-b]pyridazine-2,4(1H,3H)-dione
##STR00075##
[0676] Step 1:
(1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4-tetrahydro-pyrrolo[3,4-d]pyrimid-
in-6-yl)-carbamic acid benzyl ester
[0677]
5-Benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
(Intermediate C) (1.15 g, 3.41 mmol), benzyl carbazate (commercial)
(2.83 g, 17.05 mmol) and triethylamine (1.426 ml, 10.23 mmol) were
combined in EtOH (15 ml) and the mixture heated at reflux for 15
mins. The reaction mixture was cooled to RT and evaporated under
reduced pressure. The residue was partitioned between water and DCM
and the phases separated. The organic phase was passed through a
hydrophobic frit and evaporated under reduced pressure. The residue
was triturated with Et.sub.2O and cooled in ice until precipitation
occurred. The precipitate was collected by filtration, affording
the title compound as a white solid.
[0678] 1H NMR (400 MHz, DMSO-d6) .delta. 11.16 (br s, 1H),
7.50-7.30 (m, 9H), 7.24 (br s, 1H), 7.09 (1H, br s), 5.11 (s, 2H),
3.19 (s, 3H), other 3H singlet obscured by water signal.
[0679] LC-MS Rt 1.00 mins; ES+m/z 405 [M+H].sup.+ (Method
2minLowpH)
Step 2:
(1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4-tetrahydro-pyrrolo[3,4-d]-
pyrimidin-6-yl)-(3-hydroxy-propyl)-carbamic acid benzyl ester
[0680] 3-Bromopropan-1-ol (0.645 ml, 7.14 mmol) was added to a
suspension of
(1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4-tetrahydro-pyrrolo[3,4-d]pyri-
midin-6-yl)-carbamic acid benzyl ester (step 1) (2.22 g, 5.49
mmol), K.sub.2CO.sub.3 (2.276 g, 16.47 mmol) and
benzyltrimethylammonium chloride (0.102 g, 0.549 mmol) in
acetonitrile (50 ml). The mixture was heated to 60.degree. C. and
stirred for 40 mins. The reaction mixture was cooled to RT and
diluted with water. The mixture was extracted with EtOAc (2.times.)
and the combined organic phases washed with brine, dried over
sodium sulphate and evaporated under reduced pressure. Purification
by chromatography on silica, eluting with EtOAc/hexane and then
MeOH/EtOAc afforded the title compound as a white foam.
[0681] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.34-7.18 (m, 10H),
6.32 (s, 1H), 5.24 (d, 1H), 5.14 (d, 1H), 3.61 (dt, 2H), 3.35 (m,
1H), 3.31 (s, 3H), 3.29 (m, 1H, partially obscured), 3.27 (s, 3H),
1.50 (m, 1H), 1.35 (m, 1H).
[0682] LC-MS Rt 0.98 mins; ES+m/z 463.4 [M+H].sup.+ (Method
2minLowpH).
Step 3:
3-[Benzyloxycarbonyl-(1,3-dimethyl-2,4-dioxo-5-phenyl-1,2,3,4-tetr-
ahydro-pyrrolo[3,4-d]pyrimidin-6-yl)-amino]-propionic acid
[0683] Tetrapropylammonium perruthenate (0.152 g, 0.432 mmol) was
added to a solution of
(1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4-tetrahydro-pyrrolo[3,4-d]pyrimid-
in-6-yl)-(3-hydroxy-propyl)-carbamic acid benzyl ester (step 2) (2
g, 4.32 mmol) and N-methylmorpholine oxide monohydrate (commercial)
(2.92 g, 21.62 mmol) in Acetonitrile (40 ml). The mixture was
stirred at room temp. for 30mins. The reaction mixture was quenched
with isopropanol (50 mL) and stirred for 20mins. The mixture was
then evaporated under reduced pressure. Purification of the residue
by chromatography on silica, eluting with EtOAc followed by 1%
acetic acid/EtOAc afforded the title compound as a white solid.
[0684] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.45-7.30 (m, 10H),
6.43 (s, 1H), 5.30 (m, 2H), 3.93 (m, 1H), 3.47 (m, 1H), 3.40 (s,
3H), 3.3 (s, 3H), 2.45 (m, 1H), 2.33 (m, 1H).
[0685] LC-MS Rt 0.98 mins; ES+m/z 477.4 [M+H].sup.+ (Method
2minLowpH)
Step 4:
1,3-Dimethyl-5-phenyl-8,9-dihydropyrimido[4',5':3,4]pyrrolo[1,2-b]-
pyridazine-2,4,10(1H,3H,7H)-trione
[0686]
3-[Benzyloxycarbonyl-(1,3-dimethyl-2,4-dioxo-5-phenyl-1,2,3,4-tetra-
hydro-pyrrolo[3,4-d]pyrimidin-6-yl)-amino]-propionic acid (step 3)
(1.09 g, 2.288 mmol) and Polyphosphoric acid (3 g, 2.288 mmol) were
heated together at 100.degree. C. (block temp) for 30 mins. The
reaction mixture was cooled to RT and the residue dissolved in
minimum volume of water. The mixture was basified by slow addition
of 2M NaOH(aq), and extracted with chloroform (4.times.). The
combined organic extracts were passed through a hydrophobic frit
and evaporated under reduced pressure. Purification of the residue
by chromatography on silica, eluting with MeOH/DCM afforded the
title compound as a white solid.
[0687] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.51-7.46 (m, 2H),
7.46-7.41 (m, 3H), 4.67 (t, 1H), 3.88 (s, 3H), 3.53 (dt, 2H), 3.29
(s, 3H), 2.68 (t, 2H).
[0688] LC-MS Rt 0.83 mins; ES+m/z ion not evident [M+H].sup.+
(Method 2minLowpH).
Step 5: Trifluoro-methanesulfonic acid
5,7-dimethyl-6,8-dioxo-9-phenyl-1,2,5,6,7,8-hexahydro-1,5,7,9a-tetraaza-f-
luoren-4-yl ester
[0689] Trifluoromethanesulfonic anhydride (0.254 ml, 1.506 mmol)
was added slowly to a solution of
1,3-dimethyl-5-phenyl-8,9-dihydropyrimido[4',5':3,4]pyrrolo[1,2-b]pyridaz-
ine-2,4,10(1H,3H,7H)-trione (step 4) (222 mg, 0.684 mmol) and
2,6-lutidine (commercial) (0.159 ml, 1.369 mmol) in DCM (10 ml) at
0.degree. C. and the mixture stirred for 1.75 hours. The reaction
mixture was quenched with sat. NaHCO.sub.3(aq) and extracted with
DCM. The organic phase was passed through a hydrophobic frit and
evaporated under reduced pressure. Purification of the residue by
chromatography on silica, eluting with 20% EtOAc/hexane then 40%
EtOAc/hexane afforded the title compound as a yellow solid.
[0690] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.64-7.58 (m, 2H),
7.54-7.46 (m, 3H), 6.16 (m, 1H), 5.35 (m, 1H), 3.98 (m, 1H), 3.66
(s, 3H), 3.42 (s, 3H).
[0691] LC-MS Rt 1.07 mins; ES+m/z 499.3 [M+H+ MeCN].sup.+ (Method
2minLowpH)
Step 6:
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydropyrimid-
o[4',5':3,4]pyrrolo[1,2-b]pyridazine-2,4(1H,3H)-dione
[0692] Trifluoro-methanesulfonic acid
5,7-dimethyl-6,8-dioxo-9-phenyl-1,2,5,6,7,8-hexahydro-1,5,7,9a-tetraaza-f-
luoren-4-yl ester (step 5) (280 mg, 0.614 mmol), LiCl (2.60 mg,
0.061 mmol), copper iodide (11.68 mg, 0.061 mmol) and
4-methyl-2-tributylstannanyl-thiazole (Intermediate J) (4.45 g,
11.46 mmol) were combined in THF (15 ml) and PdCl.sub.2(dppf) (44.9
mg, 0.061 mmol) was added. The mixture was heated at reflux for 1
hour. The reaction mixture was cooled to RT and diluted with EtOAc.
The mixture was washed with dil. NH.sub.4OH(aq), 1M KF(aq) and
brine. The organic phase was dried over sodium sulphate and
evaporated under reduced pressure. Purification of the residue by
chromatography on silica, eluting with EtOAc/hexane, afforded the
title compound as a crude pale brown solid.
[0693] LC-MS: Rt 0.97 mins; ES+m/z 406.4 [M+H].sup.+ (Method
2minLowpH).
Step 7:
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8,9,10-tetrahydr-
opyrimido[4',5':3,4]pyrrolo[1,2-b]pyridazine-2,4(1H,3H)-dione
[0694]
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8-dihydropyrimido-
[4',5':3,4]pyrrolo[1,2-b]pyridazine-2,4(1H,3H)-dione (step 6) (100
mg, 0.247 mmol), 10% wt. palladium on carbon (26.2 mg, 0.025 mmol)
and ammonium formate (156 mg, 2.466 mmol) were combined in EtOH (10
ml) and the mixture heated at reflux for 24 hours. The reaction
mixture was cooled to RT and filtered through a Celite.RTM. (filter
material) cartridge (10 g), rinsing the residue with copious MeOH.
The filtrates were evaporated under reduced pressure. The residue
was triturated with EtOAc/hexane and the solid collected by
filtration. Purification of the residue by chromatography on
silica, eluting with EtOAc/hexane and MeOH/EtOAc yielded a crude
material which was further purified by preparative HPLC using the
following conditions:
[0695] Column: Waters Sunfire C18, 150 mm.times.30 mm, 5 .mu.m
[0696] Mobile Phase: A=0.1% TFA in water; B=0.1% TFA in
acetonitrile
[0697] Gradient: 0.0 min-0.5 min 30% B 30 mL/min, 0.5-1.0 min 30% B
30-50 mL/min, 1.0-7.25 min 30-70% B 50 mL/min, 7.25-7.3 70-98% B 50
mL/min, 7.3-8.3 min 98% B 50 mL/min, 8.3-8.5 min 98-30% B 50
mL/min
[0698] Detection: UV @ 220 nm
[0699] Flow rate: 10 mL/min
[0700] Injection volume: 200 .mu.l
[0701] The title compound was obtained as an off-white solid
[0702] 1H NMR (400 MHz, CDCl.sub.3) .delta. 8.26 (2H, br s),
7.62-7.57 (2H, mult), 7.52-7.43 (3H, mult), 6.96 (1H, d), 5.53 (1H,
dd), 3.41 (3H, s), 3.37 (3H, s), 2.53 (3H, d), 2.50-2.42 (2H,
mult).
[0703] LC-MS Rt 0.93 mins; ES+m/z 408.3 [M+H].sup.+ (Method
2minLowpH).
[0704] Chiral separation of the racemate by Supercritical Fluid
Chromatography was carried out using the following conditions to
afford the compound listed hereinafter:
[0705] Column: Chiralpak IB, 250.times.10 mm, 5 um;
[0706] Mobile Phase: 50% MeOH with 0.1% DEA/50% CO2
[0707] Flow Rate: 10 mL/min
Example 10.1
Enantiomer 1 of
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimi-
do[4',5':3,4]pyrrolo[1,2-b]pyridazine-2,4(1H,3H)-dione
[0708] SFC Retention time=2.76 mins
[0709] LC-MS:Rt 1.00 mins; MS 408.5 m/z [M+H] Method
2minLowpHv01
[0710] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.62 (2H, d), 7.51-7.39
(3H, m), 6.79 (1H, d), 5.13 (1H, dd), 5.09 (1H, t), 3.49 (3H, s),
3.40 (1H, m), 3.36 (3H, s), 3.28 (1H, m), 2.54 (1H, m), 2.44 (3H,
d), 2.36 (1H, m).
Example 10.2
5-(3-Chlorophenyl)-1,3-dimethyl-10-(4-methylthiazol-2-yl)-7,8,9,10-tetrahy-
dropyrimido[4',5':3,4]pyrrolo[1,2-b]pyridazine-2,4(1H,3H)-dione
##STR00076##
[0712] The title compound was prepared analogously to Example 10 by
replacing Intermediate C with the appropriate starting compound
(prepared by a similar method to Intermediate C using the
appropriate 3-chloro-benzoyl chloride in Step 2);
[0713] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.61 (1H, m), 7.53 (1H,
m), 7.40 (2H, m), 6.81 (1H, d), 5.17 (1H, dd), 3.48 (3H, s), 3.41
(1H, m), 3.37 (3H, s), 3.30 (1H, m), 2.56 (1H, m), 2.46 (3H, s),
2.38 (1H, m)
[0714] LC-MS Rt 1.16 mins; MS m/z 442.2 [M+H]+ (Method
2minLowpHv02)
Example 11.0
1,3-Dimethyl-9-(5-methylfuran-2-yl)-5-phenyl-8,9-dihydro-1H-pyrimido[4,5-a-
]pyrrolizine-2,4(3H,7H)-dione
##STR00077##
[0715] Step 1: Methyl
3-(1,3-dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-
-6(2H)-yl)propanoate
[0716]
5-Benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
(Intermediate C) (3 g, 8.90 mmol), beta alanine methyl ester
hydrochloride (1.242 g, 8.90 mmol) and triethylamine (2.480 mL,
17.80 mmol) in EtOH (63.6 mL) was heated to 100.degree. C. for 90
min using microwave radiation. The reaction mixture was diluted
with of DCM (80 ml) and washed with water and 0.1M HCl. The organic
portion was dried by passing through a phase separating cartridge
and concentrated in vacuo to yield the title compound as a pale
yellow oil.
[0717] LCMS: Rt 0.98 min; MS 342.6 [M+H]; (Method: 2minlowpH)
Step 2:
3-(1,3-Dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]py-
rimidin-6(2H)-yl)propanoic acid
[0718] A mixture of methyl
3-(1,3-dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-
-6(2H)-yl)propanoate (step 2) (3.24 g, 9.49 mmol) in THF (33.9
mL)/water (33.9 mL) and LiOH (2.273 g, 95 mmol) was heated to
50.degree. C. for 1 hr. The reaction mixture was diluted with EtOAc
(50 ml) and the layers separated. The organics were then washed
with water (3.times.50 ml). The aqueous extracts were combined and
the pH adjusted to approx. pH3 with 5M HCl. The resulting
precipitate was isolated using suction filtration.
[0719] LC-MS: Rt 0.87 mins; MS 328.4 m/z [M+H]+ (Method
2minLowpHv01)
[0720] 1H NMR: (400 MHz, CDCl.sub.3) .delta. 7.47 (5H, mult), 7.00
(1H, s), 4.09 (2H, t), 3.15 (3H, t), 2.70 (2H, t), 2.51 (3H,
s).
Step 3:
1,3-Dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,-
4,9(3H)-trione
[0721]
3-(1,3-Dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyr-
imidin-6(2H)-yl)propanoic acid (step 2) (2.11 g, 6.63 mmol) and
propylphosphonic anhydride (4.34 g, 6.63 mmol) 50% in DMF was
heated to 100.degree. C. overnight. The reaction mixture was cooled
to RT and the precipitate isolated using suction filtration. The
isolated orange solid was washed with Et.sub.2O and dried in a
vacuum oven at 50.degree. C.
[0722] 1H NMR: (400 MHz, CDCl.sub.3) .delta. 7.61 (2H, mult), 7.54
(3H, mult), 4.34 (2H, t), 3.94 (3H, s), 3.41 (3H, s), 3.17 (3H,
t).
[0723] LC-MS: Rt 0.96 mins; MS 310.2 m/z [M+H]+ (Method
2minLowpHv01)
Step 4:
1,3-Dimethyl-2,4-dioxo-5-phenyl-2,3,4,7-tetrahydro-1H-pyrimido[4,5-
-a]pyrrolizin-9-yl trifluoromethanesulfonate
[0724]
1,3-Dimethyl-5-phenyl-7,8-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4-
,9(3H)-trione (step 3) (200 mg, 0.647 mmol) and 2,6-lutidine (0.113
ml, 0.970 mmol) in DCM (2 ml) were combined under nitrogen then
cooled to 0.degree. C. in a ice bath, and trifluoromethanesulfonic
anhydride (219 mg, 0.776 mmol) was added dropwise over 5 mins. The
reaction mixture and was allowed to stir at 0.degree. C. for 35
mins then quenched with 10 ml of water. The aqueous was extracted
with DCM (2.times.10 ml). The organics were combined and dried by
passing through a phase separating cartridge. The crude product was
concentrated under vacuum. The crude product was loaded onto a 40 g
silica cartridge and the product eluted at 35% EtOAc/i-Hexane. The
relevant fractions were combined and concentrated to yield a yellow
gum. Diethyl ether was added to the gum and the mixture
concentrated once more to yield the title compound as a yellow
solid.
[0725] LC-MS: Rt 1.29 min; MS 443.2 m/z [M+H].sup.+; (Method
2minLowpH)
[0726] .sup.1H NMR: av70431 (400 MHz, DMSO) .delta. 7.76 (2H, d),
7.49 (3H, mult), 6.36 (1H, s), 5.00 (2H, d), 3.48 (3H, s), 3.22
(3H, s).
[0727] .sup.19F NMR (400 MHz, DMSO) .delta. -66.3 (CF.sub.3).
Step 5:
1,3-Dimethyl-9-(5-methylfuran-2-yl)-5-phenyl-1H-pyrimido[4,5-a]pyr-
rolizine-2,4(3H,7H)-dione
[0728]
1,3-Dimethyl-2,4-dioxo-5-phenyl-2,3,4,7-tetrahydro-1H-pyrimido[4,5--
a]pyrrolizin-9-yl trifluoromethanesulfonate (step 4) (100 mg, 0.227
mmol), CuI (4.31 mg, 0.023 mmol), lithium bromide (39.4 mg, 0.453
mmol), tributyl(5-methylfuran-2-yl)stannane (168 mg, 0.453 mmol) in
THF (2 ml) and Pd-118 (14.77 mg, 0.023 mmol) were heated to reflux
for 10mins. The reaction mixture was filtered through a glass
filter paper and the filtrate concentrated in vacuo. The crude
product was dissolved in the minimum amount DCM/EtOAc and loaded
onto a 12 g silica cartridge and eluted with 0-80% EtOAc in
iso-hexane. The desired product was eluted at 30% EtOAc. The
relevant fractions were combined and concentrated to yield a yellow
solid. The isolated solid was suspended in Et.sub.2O and the solid
filtered to yield the title product.
[0729] LC-MS: Rt 1.30mins; MS 374.3 m/z [M+H]+ (Method
2minLowpHv01)
[0730] 1H NMR: (400 MHz, CDCl.sub.3) .delta. 7.68 (2H, d), 7.51
(2H, t), 7.43 (1H, t), 6.44 (1H, d), 6.16 (1H, t), 6.09 (1H, d),
4.71 (2H, d), 3.40 (3H, s), 3.19 (3H, s), 2.37 (3H, s).
Step 6:
1,3-Dimethyl-9-(5-methylfuran-2-yl)-5-phenyl-8,9-dihydro-1H-pyrimi-
do[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[0731] To a mixture of ammonium formate (45.6 mg, 0.723 mmol),
1,3-dimethyl-9-(5-methylfuran-2-yl)-5-phenyl-1H-pyrimido[4,5-a]pyrrolizin-
e-2,4(3H,7H)-dione (step 5) (27 mg, 0.072 mmol), and Pd/C (7.69 mg,
7.23 .mu.mol) with a chip of dry ice was added ethanol (1446
.mu.L). The reaction mixture was heated to 60.degree. C. for 25
mins and allowed to stand at RT overnight. The reaction mixture was
diluted with DCM and filtered through glass filter paper. The
filtrate was washed with water and the organics collected and dried
by passing through a phase separator cartridge. The organics were
concentrated in vacuo. The crude product was loaded onto a 4 g
silica cartridge and purified using an ISCO. The product was eluted
with 0-80% EtOAc in iso-hexanes to afford the title compound;
[0732] LC-MS: Rt 1.22 mins; MS m/z 376.4 [M+H]+ (Method
2minLowpHv01)
[0733] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.63 (2H, d), 7.47 (2H,
t), 7.40 (1H, t), 5.89 (2H, mult), 4.66 (1H, mult), 4.19 (1H,
mult), 4.06 (1H, mult), 3.43 (3H, s), 3.40 (3H, s), 2.89 (1H,
mult), 2.69 (1H, mult), 2.30 (3H, s).
[0734] Chiral separation of the racemate by Supercritical Fluid
Chromatography was carried out using the following conditions to
afford the compound listed hereinafter:
[0735] Column: Chiralpak IB, 250.times.10 mm, 5 um
[0736] Mobile Phase: 25% MeOH with 0.1% DEA/75% CO.sub.2
[0737] Flow Rate: 10 mL/min
Example 11.1
Enantiomer 1 of
1,3-dimethyl-9-(5-methylfuran-2-yl)-5-phenyl-8,9-dihydro-1H-pyrimido[4,5--
a]pyrrolizine-2,4(3H,7H)-dione
[0738] SFC Retention time=5.83 mins
[0739] LC-MS: Rt 1.23 mins; MS 376.9 m/z [M+H]+ (Method
2minLowpHv01)
[0740] 1H NMR: (400 MHz, CDCl3) .delta. 7.63 (2H, d), 7.48 (2H, t),
7.40 (1H, t), 5.89 (2H, s), 4.66 (1H, d), 4.20 (1H, mult), 4.06
(1H, mult), 3.43 (3H, s), 3.40 (3H, s), 2.89 (1H, mult), 2.69 (1H,
mult), 2.29 (3H, s).
Example 12.0
10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-phenyl-7,8-dihydr-
o-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00078##
[0741] Step 1:
6-(2,3-Dihydroxypropyl)-1,3-dimethyl-5-phenyl-1H-pyrrolo[3,4-d]pyrimidine-
-2,4(3H,6H)-dione
[0742]
5-Benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
(Intermediate C)(670 mg, 1.987 mmol), 3-aminopropane-1,2-diol
(0.231 ml, 2.98 mmol) and TEA (0.554 ml, 3.97 mmol) were combined
in EtOH (15 ml) and heated at reflux for 15 mins.
[0743] The reaction mixture was cooled to RT. and evaporated under
vacuum. The residue was partitioned between DCM and diluted HCl(aq)
and the phases separated. The organic phase was passed through a
hydrophobic frit and evaporated under vacuum to afford the title
compound;
[0744] LC-MS: MS Rt 0.76 mins [M+H].sup.+ 330.3 Method
2minLowpHv01
Step 2:
6-(3-(tert-Butyldimethylsilyloxy)-2-hydroxypropyl)-1,3-dimethyl-5--
phenyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[0745]
6-(2,3-Dihydroxypropyl)-1,3-dimethyl-5-phenyl-1H-pyrrolo[3,4-d]pyri-
midine-2,4(3H,6H)-dione (step 1)(1.66 g, 5.04 mmol), imidazole
(0.686 g, 10.08 mmol) and DMAP (0.062 g, 0.504 mmol) were dissolved
in DMF (25 ml) and TBS-Cl (0.836 g, 5.54 mmol) was added. The
resulting mixture was stirred at RT overnight. Further TBS-Cl
(0.836 g, 5.54 mmol) added and stirring continued for 2 hours. The
reaction mixture was diluted with EtOAc and washed with sat.
NaHCO3(aq) and brine (3.times.). The organic phase was dried over
sodium sulphate and evaporated under vacuum. The residue was
redissolved in DCM and evaporated onto silica. The silica was
deposited onto a 25 g silica cartridge and the system eluted with
20% EtOAc/hexane, 40% EtOAc/hexane and 60% EtOAc/hexane.
[0746] The product fractions were combined and evaporated to give
tacky yellow solid. This solid was triturated with Et.sub.2O/hexane
to afford the title compound;
[0747] LC-MS: MS Rt 1.37 mins [M+H].sup.+ 444.4 Method
2minLowpHv01
Step 3:
1-((tert-Butyldimethylsilyl)oxy)-3-(1,3-dimethyl-2,4-dioxo-5-pheny-
l-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)propan-2-yl
methanesulfonate
[0748] To a solution of
6-(3-(tert-butyldimethylsilyloxy)-2-hydroxypropyl)-1,3-dimethyl-5-phenyl--
1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (step 2) (1 g, 2.25
mmol) in DCE (20 ml) was added triethylamine (1.57 ml, 11.27 mmol),
DMAP (28 mg, 0.025 mmol) and methanesulfonyl chloride (0.89 ml,
11.3 mmol) at 0.degree. C. and reaction stirred for 2 hours at RT.
Solid K.sub.2CO.sub.3 was added to the reaction mixture and then it
was diluted with water and extracted with DCM (3.times.20 ml). The
organics were passed through a hydrophobic frit and concentrated in
vacuo to yield the title compound as a brown oil which was used
without further purification;
[0749] LC-MS: Rt 1.40 mins; MS m/z 522 [M+H]+; Method
2minLowpHv01
Step 4:
S-(1-((tert-Butyldimethylsilyl)oxy)-3-(1,3-dimethyl-2,4-dioxo-5-ph-
enyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)propan-2-yl)ethanethi-
oate
[0750]
1-((tert-Butyldimethylsilyl)oxy)-3-(1,3-dimethyl-2,4-dioxo-5-phenyl-
-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)propan-2-yl
methanesulfonate (step 3)(1.3 g, 2.63 mmol) in dry DMF (20 ml) was
treated with potassium thioacetate (1.5 g, 13.1 mmol). The
resulting solution stirred for 7 h at 70.degree. C. and at RT for 3
days. The mixture was heated at 70.degree. C. overnight and after
cooling to RT, the mixture was partitioned between EtOAc (100 ml)
and water (150 ml). The layers were separated and the aqueous was
extracted with EtOAc (.times.3). The organics were combined, washed
with water, brine (.times.3), passed through a hydrophobic frit and
concentrated in vacuo to yield the title compound as a brown
oil;
[0751] LCMS: Rt 1.52 mins; MS m/z 502 [M+H]+; Method
2minLowpHv01;
Step 5:
6-(3-Hydroxy-2-mercaptopropyl)-1,3-dimethyl-5-phenyl-1H-pyrrolo[3,-
4-d]pyrimidine-2,4(3H,6H)-dione
[0752] To a stirred solution of
S-(1-((tert-butyldimethylsilyl)oxy)-3-(1,3-dimethyl-2,4-dioxo-5-phenyl-3,-
4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)propan-2-yl)ethanethioate
(step 4) (1.62 g, 3.23 mmol) in EtOH (25 ml) was added sodium
borohydride (0.61 g, 16.14 mmol) at 0.degree. C. and the solution
stirred overnight at RT. The reaction mixture was added slowly to
TFA in water (20 ml, 50%). The resulting solid was removed by
filtration and the filtrate was concentrated in vacuo to remove
EtOH. The resulting mixture was diluted with water and extracted
with DCM (.times.3). The combined organic extracts were passed
through a hydrophobic frit and concentrated under reduced pressure
to afford an oil. The oil was dissolved in a minimal volume of DCM
and purified via ISCO column chromatography, 12 g, liquid load,
0-90% EtOAc in iso-hexane to afford the title compound;
[0753] LC-MS: Rt 0.90, 0.92 mins; MS m/z 346 [M+H]+; Method
2minLowpHv01;
Step 6:
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-phenyl-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one
[0754]
6-(3-Hydroxy-2-mercaptopropyl)-1,3-dimethyl-5-phenyl-1H-pyrrolo[3,4-
-d]pyrimidine-2,4(3H,6H)-dione (step 5)(270 mg 0.78 mmol) was added
to a microwave vial (2-5 mL) equipped with a stirrer bar. To the
vial was added toluene (3 ml) followed by bismuth triflate (48.8
mg, 0.078 mmol) and 5-methylfuran-2-carbaldehyde (112 mg, 0.78
mmol). The vial was then sealed and heated to 100.degree. C. in a
microwave reactor for 30 min. The reaction mixture was reduced in
vacuo and diluted with EtOAc. Water was added, the layers separated
and the aqueous extracted with EtOAc. The organic extracts were
combined, dried (MgSO.sub.4), filtered and concentrated under
reduced pressure to yield an oil. The oil was redissolved in a
minimal volume of DCM and loaded onto silica. Purification by ISCO
column chromatography, 24 g silica, eluting with 0-80% EtOAc in
iso-hexane afford a diastereomeric mixture of the title compounds
(Example 12.0);
[0755] LCMS: Rt 1.13 mins; MS m/z 458 [M+H]+; Method
2minLowpHv01;
[0756] Chiral separation of the diastereomeric mixture (step 6) by
Supercritical Fluid Chromatography was carried out using the
following conditions to afford the compounds listed
hereinafter:
[0757] Column: Chiralpak AD-H, 250.times.10 mm, 5 um @ 35 deg
C.
[0758] Mobile phase: 30% Methanol/70% CO2
[0759] Flow: 10 ml/min
[0760] Detection: UV @ 220 nm
[0761] Instrument: Berger Minigram SFC1
[0762] Sample Concentration: 111 mg in 2 ml ethanol+1 ml THF (37
mg/ml)
Example 12a
Diastereoisomer 1 of
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-phenyl-7,8-dihyd-
ro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[0763] SFC Retention time=5.80 mins
[0764] LC-MS: Rt 1.12 mins; MS m/z 458 [M+H]+; Method
2minLowpHv01
[0765] 1H NMR (400 MHz, CDCl3) .delta. 7.54-7.41 (5H, m); 6.15 (1H,
d); 6.12 (1H, d); 5.78 (1H, s); 4.41 (1H, dd); 4.21 (1H, dd); 3.66
(3H, s); 3.47-3.36 (5H, m); 3.23-3.17 (1H, m).
Example 12b
Diastereoisomer 2 of
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-phenyl-7,8-dihyd-
ro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[0766] SFC Retention time=4.84 mins
[0767] LC-MS: Rt 1.11 mins; MS m/z 458 [M+H]+; Method
2minLowpHv01
[0768] 1H NMR (400 MHz, CDCl3) .delta. 7.55-7.43 (5H, m); 6.14 (1H,
d); 6.11 (1H, d); 5.79 (1H, s); 4.49 (1H, dd); 3.97 (1H, dd); 3.71
(2H, d); 3.67 (3H, s); 3.57-3.51 (1H, m); 3.37 (3H, s).
[0769] Also isolated were:
Diastereoisomer 3 of
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-phenyl-7,8-dihyd-
ro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[0770] SFC Retention time=3.98 mins
and Diastereoisomer 4 of
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-phenyl-7,8-dihyd-
ro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[0771] SFC Retention time=5.80 mins
Example 12.1a, 12.1b, 12.1c and 12.1d
(8R,10R)-10-(5-chlorofuran-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3--
dimethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4-
(3H,10H)-dione
(8R,10S)-10-(5-chlorofuran-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3--
dimethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4-
(3H,10H)-dione
(8S,10R)-10-(5-chlorofuran-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3--
dimethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4-
(3H,10H)-dione and
(8S,10S)-10-(5-chlorofuran-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3--
dimethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4-
(3H,10H)-dione
##STR00079##
[0772] Step 1: (2,2-Dimethyl-1,3-dioxolan-4-yl)methyl
trifluoromethanesulfonate
[0773] Trifluoromethylsulfonic anhydride (7.03 ml, 41.6 mmol) was
added dropwise over 15 mins to a solution of solketal (4.70 ml,
37.8 mmol) and 2,6-lutidine (5.73 ml, 49.2 mmol) in DCM (126 mL) at
0.degree. C. The mixture was stirred at 0.degree. C. for 1 hour.
The mixture was diluted with DCM (100 mL) and water (100 mL), the
phases were separated and the aqueous phase extracted with DCM
(3.times.50 mL). The combined organic extracts were dried over
sodium sulfate and evaporated under vacuum to afford the title
compound as a crude material which was used directly.
Step 2:
6-((2,2-Dimethyl-1,3-dioxolan-4-yl)methyl)-5-(3-fluorophenyl)-1,3--
dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[0774] Sodium hydride (60% in mineral oil) (234 mg, 5.86 mmol) was
added portionwise to a solution of
5-(3-fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-di-
one (Intermediate Nf) (1000 mg, 3.66 mmol) and dibenzo-18-crown-6
(132 mg, 0.366 mmol) in DMF (28.1 mL) at 0.degree. C. The solution
was warmed to room temperature and stirred for minutes, then
re-cooled to 0.degree. C.
(2,2-Dimethyl-1,3-dioxolan-4-yl)methyltrifluoromethane sulfonate
(Intermediate Sb) (1547 mg, 5.86 mmol) was added dropwise over 5
minutes. The mixture was warmed to room temperature and stirred for
3 hours. The reaction was quenched with saturated NH.sub.4Cl(aq)
(10 mL) and extracted with DCM (3.times.30 mL). The combined
organic extracts were dried over magnesium sulfate and evaporated
under vacuum to afford the title compound as a red/brown amorphous
solid.
[0775] LC-MS Rt 1.20 mins [M+H]+ 388.3 (Method 2minlowpHv03)
Step 3:
6-(2,3-Dihydroxypropyl)-5-(3-fluorophenyl)-1,3-dimethyl-1H-pyrrolo-
[3,4-d]pyrimidine-2,4(3H,6H)-dione
[0776] HCl (2 M in diethyl ether) (92 ml, 185 mmol) was added
dropwise to a solution of
6-((2,2-dimethyl-1,3-dioxolan-4-yl)methyl)-5-(3-fluorophenyl)-1,3-dimethy-
l-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (7.15 g, 18.46 mmol)
and water (6.65 g, 369 mmol) in acetonitrile (35.1 mL). The mixture
was stirred at room temperature for 30 minutes. Evaporation of the
reaction mixture under vacuum afforded the title compound.
[0777] LCMS Rt 0.85 mins [M+H]+ 348.3 (Method 2minlowpHV03)
Step 4:
6-(3-((tert-Butyldimethylsilyl)oxy)-2-hydroxypropyl)-5-(3-fluoroph-
enyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[0778] Tert-butyldimethylsilyl chloride (0.695 g, 4.61 mmol) was
added to a solution of
6-(2,3-dihydroxypropyl)-5-(3-fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]-
pyrimidine-2,4(3H,6H)-dione (1.82 g, 4.19 mmol), imidazole (0.571
g, 8.38 mmol) and DMAP (0.512 g, 4.19 mmol) in DMF (13.97 mL). The
mixture was stirred at room temperature for 3 hours, then diluted
with 10% aqueous citric acid solution (15 mL) and extracted with
DCM (3.times.30 mL). The combined organic extracts were dried over
magnesium sulfate and evaporated under vacuum. Purification by
chromatography on silica, eluting with 0-50% EtOAc/hexane, afforded
the title compound.
[0779] LC-MS Rt 1.52 mins [M+H].sup.+ 462.4 (Method
2minlowpHv03)
Step 5:
1-((tert-Butyldimethylsilyl)oxy)-3-(5-(3-fluorophenyl)-1,3-dimethy-
l-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)propan-2-yl
methanesulfonate
[0780] Methanesulfonic anhydride (0.679 g, 3.90 mmol) in
1,2-dichloroethane (3.0 mL) was added dropwise to a solution of
DMAP (0.016 g, 0.130 mmol), triethylamine (0.544 ml, 3.90 mmol) and
6-(3-((tert-butyldimethylsilyl)oxy)-2-hydroxypropyl)-5-(3-fluorophenyl)-1-
,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (0.6 g,
1.300 mmol) in 1,2-dichloroethane (10 mL) at 0.degree. C. The
mixture was warmed to room temperature and stirred for 2 hours,
then diluted with water (10 mL). The phases were separated and the
aqueous phase was extracted with DCM (3.times.10 mL). The combined
organic extracts were dried over magnesium sulfate and evaporated
under vacuum to afford the title compound.
[0781] LC-MS Rt 1.52 mins [M+H].sup.+ 540.6 (Method
2minlowpHv03)
Step 6:
(1-((tert-Butyldimethylsilyl)oxy)-3-(5-(3-fluorophenyl)-1,3-dimeth-
yl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)propan-2-yl)e-
thanethioate
[0782] A solution of
1-((tert-butyldimethylsilyl)oxy)-3-(5-(3-fluorophenyl)-1,3-dimethyl-2,4-d-
ioxo-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)propan-2-yl
methanesulfonate (705 mg, 1.241 mmol) and potassium thioacetate
(709 mg, 6.20 mmol) in DMF (4964 .mu.L) was heated at 70.degree. C.
for 4 hours. Further portions of potassium thioacetate (709 mg,
6.20 mmol) were added as necessary to allow the reaction to run to
completion. The reaction mixture was cooled to room temperature,
diluted with water (15 mL) and extracted with DCM (3.times.20 mL).
The combined organic extracts were dried over magnesium sulfate and
evaporated under vacuum. Purification by chromatography on silica,
eluting with 0-50% EtOAc/hexane, afforded the title compound.
[0783] LC-MS Rt 1.73 mins [M+H].sup.+ 520.4 (2minlowpHv03)
Step 7:
6-(3-((tert-Butyldimethylsilyl)oxy)-2-mercaptopropyl)-5-(3-fluorop-
henyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[0784] Sodium borohydride (74.3 mg, 1.963 mmol) was added to a
solution of (1-((tert-butyl
dimethylsilyl)oxy)-3-(5-(3-fluorophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihyd-
ro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)propan-2-yl)ethanethioate
(340 mg, 0.654 mmol) in ethanol (6542 .mu.L) at 0.degree. C. The
mixture was warmed to room temperature and stirred for 3 hours. The
reaction was quenched at 0.degree. C. with 1M HCl(aq), stirred for
10 mins, then extracted with DCM (3.times.10 mL). The combined
organic extracts were dried over magnesium sulfate and evaporated
under vacuum to afford the title compound.
[0785] LC-MS Rt 1.68 mins [M+H].sup.+ 478.3 (Method
2minlowpHv03)
Step 8:
10-(5-Chlorofuran-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-d-
imethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(-
3H,10H)-dione
[0786] Trifluoroacetic acid (161 .mu.l, 2.094 mmol) was added to a
solution of bismuth triflate (41.2 mg, 0.063 mmol),
5-chlorofuran-2-carbaldehyde (30.1 mg, 0.230 mmol) and
6-(3-((tert-butyldimethylsilyl)oxy)-2-mercaptopropyl)-5-(3-fluorophenyl)--
1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (100 mg,
0.209 mmol) in toluene (2094 .mu.L). The mixture was stirred at
room temperature for 1 hour. The reaction mixture was diluted with
DCM (10 mL) and 1M NaOH(aq) (10 mL). The phases were separated and
the aqueous phase was extracted with DCM (3.times.10 mL). The
combined organic extracts were dried over magnesium sulfate and
evaporated under vacuum. Purification by chromatography on silica,
eluting with 0-50% EtOAc/hexane, afforded the title compound as a
mixture of diastereomers.
[0787] LC-MS Rt 1.26 mins [M+H].sup.+ 476.1 (Method
2minlowpHv03)
[0788] The diastereomeric mixture was separated by SFC
chromatographic resolution under the following conditions to afford
the listed single diastereomer compounds.
[0789] Separation conditions
[0790] Column: Chiralcel AD-H 250.times.10 mm, 5 um @ 35 deg C.
[0791] Mobile phase: 40% Methanol+0.1% v/v DEA/60% CO2
[0792] Flow: 10 ml/min
[0793] Detection: UV @ 220 nm
Example 12.1a
Diastereomer 1 of
10-(5-chlorofuran-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimethyl-
-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-
-dione
[0794] Rt=4.41 mins
[0795] 1H NMR (400 MHz, CD3OD): 7.55-7.46 (1H, m), 7.32-7.20 (3H,
m), 6.25-6.21 (2H, m), 6.04 (1H, s), 4.40-4.32 (1H, m), 4.25-4.18
(1H, m), 3.62 (3H, s), 3.47-3.21 (6H, m), 3.20-3.10 (1H, m)
[0796] LC-MS Rt 1.26 mins [M+H].sup.+ 476.1 (Method
2minlowpHv03)
Example 12.1 b
Diastereomer 2 of
10-(5-chlorofuran-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimethyl-
-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-
-dione
[0797] Rt=2.53 mins was further purified by SFC under the following
conditions;
[0798] Column: 2 coupled Chiralpak IC 250.times.10 mm, 5 um
[0799] Mobile phase: 30% Methanol+0.1% v/v DEA/70% CO2
[0800] Flow: 10 ml/min
[0801] Detection: UV @ 220 nm
[0802] Rt=23.63 mins
[0803] 1H NMR (400 MHz, CD3OD): .delta. 7.57-7.49 (1H, m),
7.30-7.22 (3H, m), 6.34 (1H, d), 6.25 (1H, d), 6.05 (1H, s), 4.58
(1H, dd), 3.89-3.80 (1H, m), 3.70-3.30 (10H, m).
[0804] LC-MS Rt 1.29 mins [M+H].sup.+ 476.2 (2minlowpHV03)
Example 12.1c
Diastereomer 3 of
10-(5-chlorofuran-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimethyl-
-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-
-dione
[0805] Rt=3.56 mins
[0806] 1H NMR (400 MHz, CD3OD). 7.55-7.46 (1H, m), 7.32-7.20 (3H,
m), 6.25-6.21 (2H, m), 6.04 (1H, s), 4.40-4.32 (1H, m), 4.25-4.18
(1H, m), 3.62 (3H, s), 3.47-3.21 (6H, m), 3.20-3.10 (1H, m)
[0807] LC-MS Rt 1.31 mins [M+H].sup.+ 476.2 (2minlowpHv03)
Example 12.1d
Diastereomer 4 of
10-(5-chlorofuran-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimethyl-
-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-
-dione
[0808] Rt=4.43 mins
[0809] 1H NMR (400 MHz, CD3OD): .delta. 7.57-7.49 (1H, m),
7.30-7.22 (3H, m), 6.34 (1H, d), 6.25 (1H, d), 6.05 (1H, s), 4.58
(1H, dd), 3.89-3.80 (1H, m), 3.70-3.30 (10H, m)
[0810] LC-MS Rt 1.29 mins [M+H].sup.+ 476.2 (Method
2minlowpHv03)
[0811] The following listed examples were prepared in a similar
manner to Example 12a-12d by replacing
5-(3-fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-di-
one (Intermediate Nf) in step 2 (except Example 12.4a-12.4c) with
3-(1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-
-yl)benzonitrile (Example 9 Step 4) and with the appropriate
aldehyde (either commercially available or Intermediate described
herein) in Step 8. The diastereomeric mixtures were separated by
SFC chromatographic resolution under the listed conditions to
afford the title compounds.
Example 12.2a-12.2c
3-((8R,10R)-10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-5-yl)benzonitrile or
3-((8S,10R)-10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-5-yl)benzonitrile or
3-((8S,10S)-10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-5-yl)benzonitrile or
3-((8R,10S)-10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-5-yl)benzonitrile
##STR00080##
[0813] Separation conditions:
[0814] Column: Chiralpak AD-H, 250.times.10 mm, 5 um @ 35.degree.
C.
[0815] Mobile phase: 35% Methanol+0.1% v/v DEA/65% CO2
[0816] Flow: 10 ml/min
[0817] Detection: UV @ 220 nm
Example 12.2a
[0818] Diastereomer 1 of
3-(10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3,4-
,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)be-
nzonitrile was further purified under the following conditions:
[0819] Column: 2.times. Phenomenex LUX-C2 coupled, 250.times.10 mm,
5 um @ 35.degree. C.
[0820] Mobile phase: 50% Methanol+0.1% v/v DEA/50% CO2
[0821] Flow: 10 ml/min
[0822] Detection: UV @ 220 nm
[0823] Rt=27.67 mins
[0824] 1H NMR (400 MHz, CDCl.sub.3): .delta. 7.82-7.73 (3H, m),
7.65-7.60 (1H, m), 6.15 (2H, app. d), 5.79 (1H, s), 4.39 (1H, dd),
4.22-4.16 (1H, m), 3.97-3.90 (1H, m), 3.68 (3H, s), 3.55-3.45 (1H,
m), 3.37 (3H, s), 3.22-3.13 (1H, m), 3.10-3.0 (1H, m)
[0825] LC-MS Rt 1.27 mins [M+H].sup.+ 483.3 (Method
2minlowpHv03)
Example 12.2b
Diastereomer 2 of
3-(10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3,4-
,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)be-
nzonitrile
[0826] Rt=3.04 mins
[0827] 1H NMR (400 MHz, CDCl.sub.3): 7.65-7.60 (1H, m), 6.15 (2H,
app. d), 5.79 (1H, s), 4.39 (1H, dd), 4.22-4.16 (1H, m), 3.97-3.90
(1H, m), 3.68 (3H, s), 3.55-3.45 (1H, m), 3.37 (3H, s), 3.22-3.13
(1H, m), 3.10-3.0 (1H, m).
[0828] LC-MS Rt 1.25 mins [M+H].sup.+ 483.3 (Method
2minlowpHv03)
Example 12.2c
Diastereomer 3 of
3-(10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3,4-
,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)be-
nzonitrile
[0829] Rt=6.47 mins
[0830] 1H NMR (400 MHz, CDCl.sub.3): .delta. 7.80-7.70 (3H, m),
7.65-7.60 (1H, m), 6.17-6.14 (2H, m), 5.79 (1H, s), 4.41-4.36 (1H,
m), 4.05-3.97 (1H, m), 3.78-3.70 (2H, m), 3.67 (3H, s), 3.57-3.47
(1H, m), 3.41-3.31 (4H, m).
[0831] LC-MS Rt 1.27 mins [M+H].sup.+ 483.2 (2minlowpHv03)
Example 12.3a-12.3d
3-((8R,10R)-10-(4-Chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-d-
ioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiaz-
in-5-yl)benzonitrile or
3-((8R,10S)-10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-d-
ioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiaz-
in-5-yl)benzonitrile or
3-((8S,10R)-10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-d-
ioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiaz-
in-5-yl)benzonitrile or
3-((8S,10S)-10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-d-
ioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiaz-
in-5-yl)benzonitrile
##STR00081##
[0833] Separation conditions:
[0834] Column: Chiralpak AD-H, 250.times.10 mm, 5 um @ 34.6.degree.
C.
[0835] Mobile phase: 22% Methanol+0.1% v/v DEA/78% CO2
[0836] Flow: 10 ml/min
[0837] Detection: UV @ 220 nm
Example 12.3a
Diastereomer 1 of
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3-
,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)-
benzonitrile
[0838] Rt=16.37 mins
[0839] 1H NMR (400 MHz, DMSO-d6): .delta.7.99 (1H, s), 7.94-7.90
(1H, m), 7.82-7.77 (2H, m), 7.68-7.63 (1H, m), 6.47 (1H, s), 5.02
(1H, dd), 4.27 (1H, dd), 3.98-3.90 (1H, m), 3.59 (3H, s), 3.50-3.43
(1H, m), 3.18 (3H, s), 3.0-2.90 (1H, m)
[0840] LC-MS Rt 1.17 mins [M+H].sup.+ 500.2 (Method
2minlowpHv03)
Example 12.3b
Diastereomer 2 of
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3-
,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)-
benzonitrile
[0841] Rt=11.37 mins
[0842] 1H NMR (400 MHz, DMSO-d6): .delta. 8.0-7.96 (1H, m),
7.94-7.90 (1H, m), 7.83-7.76 (2H, m), 7.71-7.65 (1H, m), 6.47 (1H,
s), 5.23-5.17 (1H, m), 4.43-4.36 (1H, m), 3.64 (3H, s), 3.63-3.53
(3H, m), 3.19 (3H, s)
[0843] LC-MS Rt 1.14 mins [M+H].sup.+ 500.5 (Method
2minlowpHv03)
Example 12.3c
Diastereomer 3 of
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3-
,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)-
benzonitrile
[0844] Rt=14.48 mins
[0845] 1H NMR (400 MHz, DMSO-d6): .delta. 8.0-7.96 (1H, m),
7.94-7.90 (1H, m), 7.83-7.76 (2H, m), 7.71-7.65 (1H, m), 6.47 (1H,
s), 5.23-5.17 (1H, m), 4.43-4.36 (1H, m), 3.64 (3H, s), 3.63-3.53
(3H, m), 3.19 (3H, s)
[0846] LC-MS Rt 1.16 mins [M+H].sup.+ 500.2 (2minlowpHv03)
Example 12.3d
Diastereomer 4 of
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3-
,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)-
benzonitrile
[0847] Rt=14.48 mins
[0848] LC-MS Rt 1.17 mins [M+H].sup.+ 500.2 (Method
2minlowpHv03)
[0849] 1H NMR (400 MHz, DMSO-d6): .delta.7.99 (1H, s), 7.94-7.90
(1H, m), 7.82-7.77 (2H, m), 7.68-7.63 (1H, m), 6.47 (1H, s), 5.02
(1H, dd), 4.27 (1H, dd), 3.98-3.90 (1H, m), 3.59 (3H, s), 3.50-3.43
(1H, m), 3.18 (3H, s), 3.0-2.90 (1H, m)
Example 12.4a-12.4c
(8R,10R)-10-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,-
3-dimethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2-
,4(3H,10H)-dione or
(8R,10S)-10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,-
3-dimethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2-
,4(3H,10H)-dione or
(8S,10R)-10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,-
3-dimethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2-
,4(3H,10H)-dione or
(8S,10S)-10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,-
3-dimethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2-
,4(3H,10H)-dione
##STR00082##
[0851] Separation conditions:
[0852] Column: Chiralpak IC 250.times.10 mm, 5 um
[0853] Mobile phase: 45% Isopropanol+0.1% v/v DEA/55% CO2
[0854] Flow: 10 ml/min
[0855] Detection: UV @ 220 nm
Example 12.4a
Diastereomer 1 of
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10-
H)-dione
[0856] Rt=6.99 mins was further purified under the following
conditions:
[0857] Column: Chiralpak AD-H 250.times.10 mm, 5 um
[0858] Mobile phase: 30% Methanol+0.1% v/v DEA/70% CO2
[0859] Flow: 10 ml/min
[0860] Detection: UV @ 220 nm
[0861] Rt=12.88 mins
[0862] 1H NMR (400 MHz, DMSO-d6): .delta. 7.78 (1H, s), 7.52-7.44
(1H, m), 7.37-7.26 (3H, m), 6.47 (1H, s), 5.05-4.99 (1H, m),
4.32-4.25 (1H, m), 3.96-3.91 (1H, m), 3.58 (3H, s), 3.50-3.43 (1H,
m), 3.17 (3H, s), 3.04-2.95 (1H, m)
[0863] LC-MS Rt 1.19 mins [M+H].sup.+ 493.2 (Method
2minlowpHv03)
Example 12.4b
Diastereomer 2 of
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10-
H)-dione
[0864] Rt=13.85 mins
[0865] LC-MS Rt 1.20 mins [M+H].sup.+ 493.1 (2minlowpHv03)
[0866] 1H NMR (400 MHz, DMSO-d6). .delta. 7.78 (1H, s), 7.56-7.47
(1H, m), 7.38-7.26 (3H, m), 6.46 (1H, s), 5.23-5.17 (1H, m),
4.49-4.42 (1H, m), 3.63 (3H, s), 3.61-3.48 (3H, m), 3.18 (3H,
m)
Example 12.4c
Diastereomer 3 of
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10-
H)-dione
[0867] Rt=8.69 mins
[0868] 1H NMR (400 MHz, DMSO-d6): .delta. 7.78 (1H, s), 7.52-7.44
(1H, m), 7.37-7.26 (3H, m), 6.47 (1H, s), 5.05-4.99 (1H, m),
4.32-4.25 (1H, m), 3.96-3.91 (1H, m), 3.58 (3H, s), 3.50-3.43 (1H,
m), 3.17 (3H, s), 3.04-2.95 (1H, m)
[0869] LC-MS Rt 1.20 mins [M+H].sup.+ 493.1 (Method
2minlowpHv03)
Example 13
3-(10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-1,2,3,4,7,8,9,10-octah-
ydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
##STR00083##
[0870] Step 1: Lithium
4-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)butanoate
[0871] Lithium hydroxide monohydrate (1.158 g, 27.6 mmol) was added
to a suspension of methyl
4-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)butanoate (Example 9 Step 5) (10 g, 26.3 mmol)
in THF (99 ml) and water (32.9 ml). The mixture was stirred for 2
hours and evaporated under vacuum to afford the title compound.
[0872] LC-MS Rt 0.94 mins; [M-Li+H].sup.+ 367.4; (Method
2minlowpHv03)
Step 2:
4-(5-(3-Cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo-
[3,4-d]pyrimidin-6(2H)-yl)-N-methoxy-N-methylbutanamide
[0873] T3P.RTM. (31.3 ml, 52.6 mmol) was added dropwise to a
solution of N,O-dimethyllhydroxylamine hydrochloride (2.70 g, 27.6
mmol), DIPEA (18.39 ml, 105 mmol) and lithium
4-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)butanoate (9.8 g, 26.3 mmol) in DMF (263 mL).
The mixture was stirred at room temperature for 40 minutes and
quenched with water (250 mL). The resultant precipitate was
collected by filtration under reduced pressure to afford the title
compound.
[0874] LC-MS Rt 1.03 mins [M+H].sup.+ 410.5 (2minlowpHv03)
Step 3:
3-(6-(4-(4-Chlorothiazol-2-yl)-4-oxobutyl)-1,3-dimethyl-2,4-dioxo--
2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
[0875] Isopropylmagnesium chloride lithium chloride complex
solution (1.3 M in THF, 2.067 ml, 2.69 mmol) was added dropwise to
a solution of
4-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)-N-methoxy-N-methylbutanamide (1 g, 2.442 mmol)
and 2-bromo-4-chlorothiazole (Intermediate Q, step 2) (0.533 g,
2.69 mmol) in THF (24.42 mL) at 0.degree. C. The mixture was
stirred at 0.degree. C. for 30 minutes. A further portion of
isopropylmagnesium chloride lithium chloride complex solution (1.3
M in THF, 2.067 ml, 2.69 mmol) was added at 0.degree. C. and the
mixture was stirred for a further 30 minutes at 0.degree. C. The
reaction was quenched with saturated NH.sub.4Cl(aq) (20 mL) and
extracted with DCM (3.times.20 mL). The combined organic extracts
were washed with saturated NaHCO.sub.3(aq) (10 mL), dried over
magnesium sulfate and evaporated under reduced pressure.
Trituration with methanol afforded the title compound.
[0876] LC-MS Rt 1.26 mins [M+H].sup.+ 468.4 2minlowpHv03.
Step 4:
3-(6-(4-(4-Chlorothiazol-2-yl)-4-hydroxybutyl)-1,3-dimethyl-2,4-di-
oxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
[0877] Sodium borohydride (0.606 g, 16.03 mmol) was added
portionwise to a suspension of
3-(6-(4-(4-chlorothiazol-2-yl)-4-oxobutyl)-1,3-dimethyl-2,4-dioxo-2,3,4,6-
-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile (2.5 g,
5.34 mmol) in ethanol (53.4 mL) at 0.degree. C. The reaction
mixture was stirred for 15 minutes at 0.degree. C., the quenched
with water (25 mL) and extracted with DCM (3.times.50 mL). The
combined organic fractions were dried over magnesium sulfate and
evaporated under reduced pressure to afford the title compound.
[0878] LC-MS Rt 1.15 mins [M+H].sup.+ 470.4 (2minlowpHv03).
Step 5:
3-(10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-1,2,3,4,7,8,9,-
10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
[0879] Trifluoromethanesulfonic anhydride (3.95 ml, 23.41 mmol) was
added dropwise to a solution of
3-(6-(4-(4-chlorothiazol-2-yl)-4-hydroxybutyl)-1,3-dimethyl-2,4-dioxo-2,3-
,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile (5 g,
10.64 mmol) and triethylamine (3.56 ml, 25.5 mmol) in DCM (213 mL)
at -78.degree. C. The mixture was stirred for 1 min at -78.degree.
C. The reaction was quenched with saturated aqueous NaHCO.sub.3(aq)
(100 mL) and extracted with DCM (3.times.100 mL). The combined
organic extracts were dried over magnesium sulfate and evaporated
under reduced pressure to afford crude racemic
3-(10-(4-chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-1,2,3,4,7,8,9,10-octa-
hydropyrimido[4,5-a]indolizin-5-yl)benzonitrile.
[0880] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.80-7.72 (3H, mult),
7.63 (1H, t), 7.05 (1H, s), 5.17 (1H, mult), 4.05 (1H, mult), 3.79
(1H, mult), 3.44 (6H, br s), 2.63 (1H, mult), 2.34 (1H, mult),
1.98-1.84 (2H, mult).
[0881] LC-MS Rt 1.15 mins [M+H].sup.+ 452.4 (Cl isotopes)(Method
2minLowpHv01)
[0882] Chiral separation of the racemate by Supercritical Fluid
Chromatography was carried out using the following conditions to
afford the compounds listed hereinafter:
Example 13a
(R)-3-(10-(4-chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-1,2,3,4,7,8,9,10-o-
ctahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
##STR00084##
[0884] Column: Chiralcel OJ-H 250.times.10 mm, 5 um @ 35 C
[0885] Mobile phase: 40% Methanol/60% CO2
[0886] Flow: 10 ml/min
[0887] Detection: UV @ 220 nm
Enantiomer
(R)-3-(10-(4-chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-1,2,3,4-
,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
[0888] Rt=5.48 mins
[0889] NMR (400 MHz, DMSO-d6) .delta. 8.00 (1H, s), 7.93 (1H, d),
7.84 (1H, d), 7.68 (1H, t), 7.65 (1H, s), 5.31 (1H, mult), 3.98
(1H, mult), 3.90 (1H, mult), 3.31 (3H, s), 3.16 (3H, s), 2.36 (1H,
mult), 2.26 (1H, mult), 1.84 (1H, mult), 1.68 (1H, mult).
[0890] LC-MS Rt 1.28 mins [M+H].sup.+ 452.5 (Method
2minLowpHv03)
[0891] The other enantiomer (S) was isolated at Rt=4.54 mins
[0892] The compounds of the following examples were prepared by a
similar method to that of Example 13 and 13a from the appropriate
starting material (prepared by an analogous method to methyl
4-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)butanoate (Example 9 Step 5) using the
appropriate halo compound in Step 3) and using the appropriate halo
compound in step 3). The racemate were resolved by SFC
chromatographic according the conditions described.
Example 13.1
10-(4-Chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-7-
,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00085##
[0894] 1H NMR (400 MHz, DMSO-d6) .delta. 7.72 (1H, s), 7.71 (1H,
s), 5.50-5.38 (2H, m), 4.71 (2H, s), 4.64 (1H, d), 4.05 (1H, td),
3.38 (3H, s), 3.27 (3H, s), 2.44-2.27 (2H, m), 1.95 (1H, d), 1.76
(1H, m).
[0895] LC-MS Rt 1.02 mins [M+H].sup.+ 464.4/466.4 (Method
2minLowpHv03).
Example 13.1a
(R)-10-(4-chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimeth-
yl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
or
(S)-10-(4-chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimeth-
yl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00086##
[0897] Column: Phenomenex LUX-C4, 250.times.10 mm, 5 um @35 deg
C.
[0898] Mobile phase: 50% Methanol/50% CO2
[0899] Flow: 10 ml/min
[0900] Detection: UV @ 220 nm
Example 13.1a
Enantiomer 1 of
10-(4-chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl--
7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0901] Rt=3.25 mins
[0902] 1H NMR (400 MHz, DMSO-d6) .delta. 7.66 (1H, s), 7.65 (1H,
s), 5.39 (1H, t), 5.36 (1H, m), 4.65 (2H, d), 4.58 (1H, m), 3.99
(1H, td), 3.32 (3H, s), 3.21 (3H, s), 2.38-2.21 (2H, m), 1.89 (1H,
s), 1.69 (1H, obs q).
[0903] LC-MS Rt 1.06 mins [M+H].sup.+ 464.3/466.3 (Method
2minLowpHv03
[0904] The second enantiomer was isolated at Rt=4.20 mins
Example 13.2
1,3-Dimethyl-5,10-bis(4-methylthiazol-2-yl)-7,8,9,10-tetrahydropyrimido[4,-
5-a]indolizine-2,4(1H,3H)-dione
##STR00087##
[0906] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.02 (1H, s), 6.69 (1H,
s), 5.04 (1H, mult), 4.60 (1H, mult), 4.01 (1H, mult), 3.32 (6H,
s), 2.45 (3H, s), 2.37 (3H, s), 2.32 (1H, mult), 2.21 (1H, mult),
1.86 (2H, mult).
[0907] LC-MS Rt 1.23 mins [M+H].sup.+ 428.3 (Method
2minLowpHv03)
Example 13.3
10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8,9,10-t-
etrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00088##
[0909] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.31-7.26 (5H, mult),
7.20 (1H, s), 7.04 (1H, s), 5.15 (1H, mult), 4.67 (1H, mult), 4.24
(1H, mult), 3.43 (3H, s), 3.42 (3H, s), 2.60 (3H, s), 2.59 (1H,
mult), 2.34 (1H, mult), 2.04-1.89 (2H, mult).
[0910] LC-MS Rt 1.30 mins; [M+H].sup.+ 433.1 (Method
2minLowpHv03).
Example 13.3a
(R)-10-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8,9,-
10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione or
(S)-10-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8,9,-
10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00089##
[0912] Column: Chiralpak AD-H 250.times.10 mm, 5 um @ 35 deg C.
[0913] Mobile phase: 45% Methanol+0.1% v/v DEA/55% CO2
[0914] Flow: 10 ml/min
[0915] Detection: 220 nm
Example 13.3a
Enantiomer 1 of
10-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-7,8,9,10--
tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0916] Rt=4.34 mins
[0917] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.12 (1H, s), 7.03 (1H,
s), 5.13 (1H, dd), 4.71 (1H, mult), 4.08 (1H, mult), 3.42 (3H, s),
3.41 (3H, s), 2.61 (1H, mult), 2.54 (3H, s), 2.31 (1H, mult),
2.02-1.82 (2H, mult)
[0918] LC-MS Rt 1.27 mins [M+H].sup.+ 448.1 (Method
2minLowpHv03).
[0919] The second enantiomer was isolated at Rt=6.38 mins
Example 13.4
10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(2-methylthiazol-4-yl)-7,8,9,10-t-
etrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00090##
[0921] 1H NMR (400 MHz, DMSO-d6) .delta. 7.90 (1H, s), 7.63 (1H,
s), 5.30 (1H, m), 4.04 (1H, d), 3.87 (1H, td), 3.31 (3H, s), 3.19
(3H, s), 2.73 (3H, s), 2.36 (1H, d), 2.25 (1H, t), 1.87 (1H, d),
1.68 (1H, q).
[0922] LCMS Rt 1.24 min [M+H]+ 448.4 (Method 2minLowpHv03)
Example 13.4a
(R)-10-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-(2-methylthiazol-4-yl)-7,8,9,-
10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione or
(S)-10-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-(2-methylthiazol-4-yl)-7,8,9,-
10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00091##
[0924] Column: Chiralcel AD-H 250.times.10 mm.times.5 um @
35.degree. C.
[0925] Mobile phase: 50% Methanol/50% CO2
[0926] Flow: 10 ml/min
[0927] Detection: UV @ 220 nm
Example 13.4a
Enantiomer 1 of
10-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-(2-methylthiazol-4-yl)-7,8,9,10--
tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[0928] Rt=3.82 mins
[0929] 1H NMR (400 MHz, DMSO-d6) .delta. 7.90 (s, 1H), 7.63 (s,
1H), 5.30 (t, 1H), 4.40 (d, 1H), 3.87 (td, 1H), 3.31 (s, 3H), 3.20
(s, 3H), 2.73 (s, 3H), 2.36 (d, 1H), 2.26 (s, 1H), 1.86 (s, 1H),
1.69 (s, 1H).
[0930] LC-MS Rt 1.18 min [M+H].sup.+ 448.3 (Method
2minLowpHv04)
[0931] The second enantiomer was isolated at Rt=4.91 mins.
Example 14
(8S,10R)-10-(5-Chlorofuran-2-yl)-8-((dimethylamino)methyl)-1,3-dimethyl-5--
phenyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione or
(8R,10R)-10-(5-chlorofuran-2-yl)-8-((dimethylamino)methyl)-1,3-dimethyl-5--
phenyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione or
(8R,10S)-10-(5-chlorofuran-2-yl)-8-((dimethylamino)methyl)-1,3-dimethyl-5--
phenyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione or
(8S,10S)-10-(5-chlorofuran-2-yl)-8-((dimethylamino)methyl)-1,3-dimethyl-5--
phenyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione
##STR00092## ##STR00093##
[0933] Methanesulfonic anhydride (60.9 mg, 0.35 mmol) was added to
a solution of
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-phenyl-7,8-dihyd-
ro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
(Example 12.0) (160 mg, 0.35 mmol) and 2,6-lutidine (0.041 ml, 0.35
mmol) in DCM (16 ml) at 0.degree.. The mixture was stirred at room
temperature for 2 hours. The mixture was evaporated under vacuum
and the residue redissolved in THF (6 ml). Dimethylamine solution
(2M in THF, 6 ml, 12.0 mmol) was added and mixture stirred at room
temperature for 1 h, at 35.degree. C. for 16 hours, at 55.degree.
C. for 6 hours, again at room temperature for 3 days, then at
55.degree. C. for 4 hours, and finally heated under microwave
irradiation at 80.degree. C. for 20 mins. The mixture was
evaporated under vacuum. The residue was redissolved in DCM and
methanol and loaded onto an Isolute.TM. SCX-2 cartridge. The column
was first rinsed with methanol/DCM (1:1, 50 ml) then eluted with
3.5 M NH.sub.3/methanol (50 ml). The basic eluent was evaporated
under vacuum to afford the title compounds as a mixture of
diastereomers
[0934] LC-MS: Rt 0.75 and 0.78 mins [M+H].sup.+ 485 (Method
2minLowpHv01)
[0935] The diastereomers were separated by SFC chromatographic
resolution under the following conditions to afford the title
compounds.
[0936] Column: Chiralcel AD 250.times.10 mm 5 u @ 35 deg C.
[0937] Mobile phase: 20% Methanol+0.1% DEA/80% CO2
[0938] Flow: 10 ml/min
[0939] Detection: UV @ 220 nm
Example 14a
Diastereoisomer 1 of
10-(5-chlorofuran-2-yl)-8-((dimethylamino)methyl)-1,3-dimethyl-5-phenyl-7-
,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-d-
ione
[0940] Rt=4.85 mins
[0941] 1H NMR (400 MHz, DMSO) .delta. 7.50-7.44 (5H, m); 6.43 (1H,
d); 6.26 (1H, d); 6.14 (1H, s); 4.16-4.06 (2H, m); 3.52-3.47 (4H,
s); 3.15 (3H, s); 2.02 (2H, dd); 1.94 (6H, s).
[0942] LC-MS Rt 0.84 mins [M+H].sup.+ 485 (Method 2minLowpHv03)
Example 14b
Diastereoisomer 2 of
10-(5-chlorofuran-2-yl)-8-((dimethylamino)methyl)-1,3-dimethyl-5-phenyl-7-
,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-d-
ione
[0943] Rt=6.88 mins
[0944] 1H NMR (400 MHz, DMSO) .delta. 7.50-7.43 (5H, m); 6.42 (1H,
d); 6.32 (1H, d); 6.15 (1H, s); 4.35 (1H, dd); 3.70 (1H, dd);
3.64-3.55 (1H, m); 3.53 (3H, s); 3.15 (3H, s); 2.38 (2H, dd); 2.06
(6H, s)
[0945] LC-MS Rt 0.81 mins [M+H].sup.+ 485 (Method 2minLowpHv03)
Example 15
(R)-10-(5-Chlorofuran-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-
-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-
-dione or
(S)-10-(5-Chlorofuran-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-
-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-
-dione
##STR00094##
[0946] Step 1: Ethyl
2-(10-(5-chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-hexahydro--
1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)thiazole-4-carboxyla-
te
[0947] TFA (0.858 ml, 11.14 mmol) was added to a mixture of ethyl
2-(1,3-dimethyl-2,4-dioxo-6-(2-(tritylthio)ethyl)-2,3,4,6-tetrahydro-1H-p-
yrrolo[3,4-d]pyrimidin-5-yl)thiazole-4-carboxylate (Intermediate
GB) (473 mg, 0.743 mmol), triethylsilane (0.119 ml, 0.743 mmol) and
bismuth triflate (244 mg, 0.371 mmol) in toluene (5.4 ml). The
mixture was stirred at room temperature for 48 hours. The mixture
was diluted with saturated NaHCO.sub.3(aq) (30 ml) and extracted
with EtOAc (3.times.20 ml). The combined organic extracts were
washed with water (20 ml) and brine (20 ml), dried over sodium
sulfate and evaporated under vacuum. Purification by chromatography
on silica, eluting with 30-50% EtOAc/hexane afforded the title
compound.
[0948] 1H NMR (400 MHz, DMSO-d6) .delta. 8.73 (1H, s), 6.42 (1H,
d), 6.22 (1H, s), 6.21 (1H, d), 4.58 (1H, dt), 4.42 (1H, m), 4.38
(2H, q), 3.45 (3H, s), 3.21 (3H, s), 2.98 (1H, m), 1.33 (3H,
t).
[0949] LC-MS Rt 1.41 mins [M+H].sup.+ 507.3/509.3 (Method
2minLowpHv03)
Step 2:
rac-10-(5-Chlorofuran-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3--
dimethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4-
(3H,10H)-dione
[0950] Sodium borohydride (45.1 mg, 1.191 mmol) was added to a
solution of ethyl
2-(10-(5-chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-hexa-
hydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)thiazole-4-car-
boxylate (302 mg, 0.596 mmol) and lithium chloride (50.5 mg, 1.191
mmol) in ethanol (7.50 ml) and THF (15 ml) at 0.degree. C. The
mixture was warmed to room temperature and stirred for 7 hours.
Further portions of lithium chloride (50.5 mg, 1.191 mmol) and
sodium borohydride (45.1 mg, 1.191 mmol) were added to allow the
reaction to run to completion. The reaction was quenched with
saturated NaHCO.sub.3(aq) solution (50 ml), diluted with water (20
ml) and extracted with DCM (3.times.75 ml). The combined organic
extracts were washed with water (20 ml), passed though a
hydrophobic frit and evaporated under vacuum to afford the title
compound.
[0951] 1H NMR (400 MHz, DMSO-d6) .delta. 7.68 (1H, s), 6.42 (1H,
d), 6.20 (1H, s), 6.17 (1H, d), 5.40 (1H, t), 4.65 (2 h, d), 4.54
(1H, dt), 4.38 (1H, m), 3.45 (3H, s), 3.20 (3H, s), 3.03-2.96 (2H,
m)
[0952] LC-MS Rt 1.22 mins [M+H].sup.+ 465.3/467.3 (Method
2minLowpHv03)
Step 3:
(R)-10-(5-Chlorofuran-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3--
dimethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4-
(3H,10H)-dione
[0953] Separation of racemic
10-(5-chlorofuran-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one was carried out under the following conditions to afford the
title compound.
[0954] Column: Chiralcel OD-H 250.times.10 mm, 5 um @ 35 deg C.
[0955] Mobile phase: 50% Methanol/50% CO2
[0956] Flow: 10 ml/min
[0957] Detection: UV @ 220 nm
Enantiomer 1 of
10-(5-Chlorofuran-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one or
(S)-10-(5-Chlorofuran-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-d-
imethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(-
3H,10H)-dione
[0958] Rt=6.73 mins
[0959] 1H NMR (400 MHz, DMSO-d6) .delta. 7.68 (1H, s), 6.41 (1H,
d), 6.19 (1H, s), 6.16 (1H, dd), 5.39 (1H, t), 4.65 (2H, d), 4.53
(1H, dt), 4.38 (1H, m), 3.45 (3H, s), 3.20 (3H, s), 3.04-2.96 (2H,
m).
[0960] LC-MS Rt 1.17 mins [M+H].sup.+ 465.2 (Method
2minLowpHv03)
[0961] Chiral purity >99% e.e.
[0962] The other enantiomer was isolated at Rt=5.07 mins
[0963] The compounds of the following examples were prepared
analogously to Example 15 (steps 1 and 2) by replacing Intermediate
GB (step 1) with the appropriate starting compound (prepared by a
similar method to Intermediate GB from Intermediate F and the
appropriate halo compound in and the commercially available
aldehyde.
Example 15.1
10-(4-Chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-7-
,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-d-
ione
##STR00095##
[0965] 1H NMR (400 MHz, DMSO-d6) .delta. 7.379 (1H, s), 7.69 (1H,
s), 6.51 (1H, s), 5.39 (1H, t), 4.72-4.60 (3H, m), 4.30 (1H, m),
3.53 (3H, s), 3.22 (3H, s), 3.19-3.02 (2H, m)
[0966] LC-MS Rt 1.07 mins [M+H].sup.+ 482.0 (Method
2minLowpHv03)
Example 15.2
10-(5-Chlorofuran-2-yl)-5-(3-ethoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-pyr-
imido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00096##
[0968] LC-MS Rt 1.32mins [M+H].sup.+ 472.5 (Method
2minLowpHv01)
[0969] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.40 (1H, t), 6.99 (2H,
t), 6.94 (1H, d), 6.11 (1H, d), 5.93 (1H, d), 5.72 (1H, s), 4.21
(1H, mult), 4.10 (3H, mult), 3.59 (3H, s), 3.36 (3H, s), 3.08 (1H,
mult), 2.83 (1H, mult), 1.45 (3H, t)
Example 15.3
5-(3-Methoxyphenyl)-1,3-dimethyl-10-(4-methylthiazol-2-yl)-7,8-dihydro-1H--
pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00097##
[0971] 1H NMR (400 MHz, DMSO-d6) .delta. 7.39 (1H, t), 7.24 (1H,
s), 7.04 (1H, dd), 7.01-6.96 (2H, m), 6.37 (1H, s), 4.12 (1H, m),
4.03 (1H, m), 3.79 (3H, s), 3.52 (3H, s), 3.15 (3H, s), 3.11 (1H,
m), 3.01 (1H, m)
[0972] LC-MS Rt 1.12 mins [M+H].sup.+ 455.3 (Method
2minLowpHv01)
Example 15.4
10-(5-Chlorofuran-2-yl)-5-(3-methoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-py-
rimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00098##
[0974] 1H NMR (400 MHz, DMSO-d6) .delta. 7.38 (1H, t), 7.02 (3H,
obs t), 6.41 (1H, d), 6.15-6.11 (2H, m), 4.12 (1H, dt), 3.97 (1H,
m), 3.79 (3H, s), 3.44 (3H, s), 3.13 (3H, s), 2.98-2.92 (2H, m)
[0975] LC-MS Rt 1.26 mins [M+H].sup.+ 458.2 (Method
2minLowpHv01)
Example 15.5
10-(2-Chlorothiazol-4-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8-di-
hydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00099##
[0977] 1H NMR (400 MHz, DMSO) .delta. 7.80 (1H, s), 7.23 (1H, s),
6.69 (1H, s), 6.15 (1H, s), 4.48 (1H, mult), 4.37 (1H, mult), 3.94
(3H, s), 3.45 (3H, s), 3.18 (3H, s), 2.99 (2H, mult)
[0978] LC-MS Rt 1.13 mins [M+H].sup.+ 449.5 (Method
2minLowpHv03)
Example 15.6
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8-dihy-
dro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00100##
[0980] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.49 (1H, d), 6.81 (1H,
d), 6.09 (1H, d), 5.96 (1H, d), 5.71 (1H, s), 4.61 (1H, mult), 4.46
(1H, mult), 3.99 (3H, s), 3.58 (3H, s), 3.39 (3H, s), 3.12 (1H,
mult), 2.89 (1H, mult)
[0981] LC-MS Rt 1.11 mins [M+H].sup.+ 432.6 (Method
2minLowpHv01)
Example 15.7
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-imidazol-4-yl)-7,8-dih-
ydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00101##
[0983] 1H NMR (400 MHz, CDCl.sub.3) .delta. 8.53 (1H, s), 7.94 (1H,
s), 6.11 (1H, mult), 6.00 (1H, mult), 5.72 (1H, s), 4.66 (1H,
mult), 4.25 (1H, mult), 4.05 (3H, s), 3.59 (3H, s), 3.44 (3H, s),
3.19 (1H, mult), 3.02 (1H, mult)
[0984] LC-MS Rt 0.76 mins [M+H].sup.+ 432.6 (Method
2minLowpHv01)
Example 15.8
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-4-yl)-7,8-dihy-
dro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00102##
[0986] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.88 (1H, s), 7.64 (1H,
s), 6.08 (1H, d), 5.92 (1H, d), 5.71 (1H, s), 4.38 (1H, m), 4.26
(1H, m) 4.01 (3H, s), 3.56 (3H, s), 3.37 (3H, s), 3.08 (1H, ddd),
2.89 (1H, dt).
[0987] LC-MS Rt 0.80 mins [M+H].sup.+ 430.2 (Method
2minLowpHv02)
Example 15.9
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(5-methyl-1,3,4-oxadiazol-2-yl)-7,8-
-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dio-
ne
##STR00103##
[0989] 1H NMR (400 MHz, CDCl3) .delta. 6.12 (1H, d), 5.97 (1H,
mult), 5.72 (1H, s), 4.78 (1H, mult), 4.60 (1H, mult), 3.60 (3H,
s), 3.43 (3H, s), 3.19 (1H, mult), 2.97 (1H, mult), 2.71 (3H,
s)
[0990] LC-MS Rt 0.98 mins [M+H].sup.+ 465.3 (Method
2minLowpHv01)
Example 15.10
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(4-methyloxazol-2-yl)-7,8-dihydro-1-
H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00104##
[0992] 1H NMR (400 MHz, CD3OD+5 drops DMSO-d6) .delta. 7.68 (1H,
s), 6.25 (1H, d), 6.09 (1H, d), 6.04 (1H, s), 4.85 (1H, dt), 4.35
(1H, m), 3.55 (3H, s), 3.33 (3H, s), 3.15 (1H, m), 3.00 (1H, dt),
2.29 (3H, s)
[0993] LC-MS Rt 1.27 mins [M+H].sup.+ 433.1 (Method
2minLowpHv03)
[0994] The compounds of the following examples were prepared
analogously to Example 15a and 15b by replacing Intermediate GB
(step 1) with the appropriate starting compound (prepared by a
similar method to Intermediate GB starting from Intermediate F and
the appropriate halo compound) and the commercially available
aldehyde. The resulting racemates were separated by SFC
chromatographic resolution to yield single enantiomers.
Example 15.11
(R)-10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8--
dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dion-
e or
(S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8--
dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dion-
e
##STR00105##
[0996] Column: Chiralcel OD 250.times.10 mm 5 .mu.m@ 35.1.degree.
C.
[0997] Eluent: 50% MeOH/50% scCO2
[0998] Flow: 10 ml/min
[0999] Detection: UV @ 220 nm
Enantiomer 1 of
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8-dih-
ydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[1000] Rt=7.67 mins
[1001] LC-MS Rt 1.12 mins [M+H].sup.+ 432.1 (Method
2minLowpHv01)
[1002] 1H NMR (400 MHz, DMSO-d6) .delta. 7.79 (1H, d), 6.67 (1H,
d), 6.39 (1H, d), 6.13 (1H, s), 6.07 (1H, dd), 4.44 (1H, dt), 4.29
(1H, m), 3.91 (3H, s), 3.43 (3H, s), 3.16 (3H, s), 3.00-2.94 (2H,
m)
Example 15.12
(S)-10-(5-Chlorofuran-2-yl)-5-(3-ethoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-
-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
or
(R)-10-(5-chlorofuran-2-yl)-5-(3-ethoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-
-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00106##
[1004] Separation conditions:
[1005] Column: Chiralcel OD-3, 150.times.2.1 mm 3 um @ 40 C,
[1006] Eluent: 40% MeOH/60% CO2
[1007] Flow: 0.4 ml/min
[1008] Detection: UV @ 220 nm and 254 nm
Enantiomer 1 of
10-(5-Chlorofuran-2-yl)-5-(3-ethoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-py-
rimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[1009] Rt=6.85 mins
[1010] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.41 (1H, t), 6.98 (3H,
mult), 6.10 (1H, d), 5.93 (1H, mult), 5.72 (1H, s), 4.32 (4H,
mult), 3.59 (3H, s), 3.35 (3H, s), 3.08 (1H, mult), 2.83 (1H,
mult), 1.45 (3H, t)
Example 15.13
(S)-10-(5-Chlorofuran-2-yl)-5-(3-methoxyphenyl)-1,3-dimethyl-7,8-dihydro-1-
H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
or
(R)-10-(5-Chlorofuran-2-yl)-5-(3-methoxyphenyl)-1,3-dimethyl-7,8-dihydro-1-
H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00107##
[1012] Separation conditions:
[1013] Column: Chiralcel OD 250.times.10 mm 5 um @ 34.8.degree.
C.
[1014] Mobile phase: 35% Methanol/65% CO2
[1015] Flow: 10 ml/min
[1016] Detection: UV @ 220-260 nm
Enantiomer 1 of
10-(5-Chlorofuran-2-yl)-5-(3-methoxyphenyl)-1,3-dimethyl-7,8-dihydro-1H-p-
yrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[1017] Rt=5.40 mins
[1018] 1H NMR (400 MHz, DMSO-d6) .delta. 7.38 (1H, td), 7.07-6.98
(3H, m), 6.41 (1H, d), 6.16-6.10 (2H, m), 4.12 (1H, dt), 3.98 (1H,
m), 3.79 (3H, s), 3.44 (3H, s), 3.14 (3H, s), 2.98-2.92 (2H, m)
[1019] LC-MS Rt 1.26 mins [M+H].sup.+ 458.3 (Method
2minLowpHv01)
Example 15.14
(S)-10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(5-methyl-1,3,4-oxadiazol-2-yl)-
-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-
-dione or
(R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(5-methyl-1,3,4-oxadiazol-2-yl)-
-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-
-dione
##STR00108##
[1021] Separation conditions:
[1022] Column: Chiralpak ID, 250.times.10 mm, 5 um @ 35 deg C.
[1023] Mobile phase: 50% Methanol/50% CO2
[1024] Flow: 10 ml/min
[1025] Detection: UV @ 220 nm
Enantiomer 1 of
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(5-methyl-1,3,4-oxadiazol-2-yl)-7,-
8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)-di-
one
[1026] Rt=6.64 mins
[1027] 1H NMR (400 MHz, DMSO-d6) .delta. 6.42 (1H, d), 6.23 (1H,
s), 6.15 (1H, dd), 4.55 (1H, dt), 4.27 (1H, m), 3.43 (3H, s), 3.20
(3H, s), 3.07 (1H, dt), 2.98 (1H, m), 2.61 (3H, s)
[1028] LC-MS Rt 1.15 mins [M+H].sup.+ 434.3 (Method
2minLowpHv01)
Example 15:15
(R)-10-(5-Chlorofuran-2-yl)-5-(4-((S)-1-hydroxyethyl)thiazol-2-yl)-1,3-dim-
ethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H-
,10H)-dione or
(R)-10-(5-chlorofuran-2-yl)-5-(4-((R)-1-hydroxyethyl)thiazol-2-yl)-1,3-dim-
ethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H-
,10H)-dione or
(S)-10-(5-chlorofuran-2-yl)-5-(4-((S)-1-hydroxyethyl)thiazol-2-yl)-1,3-dim-
ethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H-
,10H)-dione or
(S)-10-(5-chlorofuran-2-yl)-5-(4-((R)-1-hydroxyethyl)thiazol-2-yl)-1,3-dim-
ethyl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H-
,10H)-dione
##STR00109##
[1030] Separation conditions:
[1031] Column: Chiralcel OJ-H 250.times.10 mm, 5 um @ 35 deg C.
[1032] Mobile phase: 30% Methanol+0.1% v/v DEA/70% CO2
[1033] Flow: 10 ml/min
[1034] Detection: UV @ 220 nm
Diastereoisomer 1 of
10-(5-Chlorofuran-2-yl)-5-(4-((S)-1-hydroxyethyl)thiazol-2-yl)-1,3-dimeth-
yl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10-
H)-dione
[1035] Rt=5.72 mins
[1036] 1H NMR (400 MHz, DMSO-d6) .delta. 7.64 (1H, s), 6.42 (1H,
d), 6.20 (1H, s), 6.18 (1H, d), 5.39 (1H, br s), 4.89 (1H, m), 4.53
(1H, dt), 4.41 (1H, m), 3.45 (3H, s), 3.20 (3H, s), 3.05-2.95 (2H,
m), 1.45 (3H, d)
[1037] LC-MS Rt 1.25 mins [M+H].sup.+ 495.2 (Method
2minLowpHv03)
Example 15.16
(R)-1-(4-Chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethy-
l-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H-
)-dione or
(S)-10-(4-chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimeth-
yl-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10-
H)-dione
##STR00110##
[1039] Separation conditions:
[1040] Column: Chiralpak IF 250.times.10 mm, 5 um @ 35.1.degree.
C.
[1041] Mobile Phase: 50% MeOH/50% CO2
[1042] Flow: 15 ml/min
[1043] Detection: UV @ 220 nm
Enantiomer 1 of
10-(4-Chlorothiazol-2-yl)-5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl--
7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,10H)--
dione
[1044] Rt=4.40 mins
[1045] 1H NMR (400 MHz, DMSO-d6) .delta. 7.78 (1H, s), 7.68 (1H,
s), 6.50 (1H, s), 5.38 (1H, t), 4.67 (1H, m), 4.63 (2H, d), 4.30
(1H, m), 3.52 (3H, s), 3.31 (3H, s), 3.14 (1H, m), 3.06 (1H,
m).
[1046] LC-MS Rt 1.08 mins [M+H].sup.+ 482.3 (Method
2minLowpHv03).
Example 16
9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9-dihydro-1H-pyrimido[4,5-
-a]pyrrolizine-2,4(3H,7H)-dione
##STR00111##
[1047] Step 1:
3-(1,3-Dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-
-6(2H)-yl)-N-methoxy-N-methylpropanamide
[1048] Isopropylmagnesium bromide solution (2.9M, 3.03 mL, 8.79
mmol) was added slowly to a suspension of methyl
3-(1,3-dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-
-6(2H)-yl)propanoate (Example 11.0 step 1) (1.5 g, 4.39 mmol) and
commercially available N,O-dimethylhydroxylamine hydrochloride
(0.514 g, 5.27 mmol) in THF (50 mL) at 0.degree. C. The mixture was
warmed to room temperature and stirred for 30 mins. A further
portion of isopropylmagnesium bromide solution (2.9M, 3.03 mL, 8.79
mmol) was added slowly and the mixture stirred for a further 60
mins. The reaction was quenched with saturated NH.sub.4Cl(aq) and
extracted with EtOAc (2.times.). The combined organic extracts were
washed with brine, dried over sodium sulfate and evaporated under
vacuum. Purification by chromatography on silica, eluting with
50-100% EtOAc/hexane, then 1-8% MeOH/EtOAc afforded the title
compound.
[1049] LC-MS Rt 0.99 mins [M+H].sup.+ 371.2 (Method
2minLowpHv03)
[1050] 1H NMR (400 MHz, CDCl3) .delta. 7.47-7.29 (5H, mult), 6.42
(1H, br s), 4.19 (2H, t), 3.45 (3H, s), 3.26 (3H, s), 3.07 (3H, s),
2.64 (2H, t).
Step 2:
6-(3-(4-Chlorothiazol-2-yl)-3-oxopropyl)-1,3-dimethyl-5-phenyl-1H--
pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1051] Isopropylmagnesium chloride lithium chloride complex
solution (1.3M, 2.93 mL, 3.81 mmol) was added dropwise to a
solution of
3-(1,3-dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-
-6(2H)-yl)-N-methoxy-N-methylpropanamide (470 mg, 1.269 mmol) and
2-bromo-4-chlorothiazole (Intermediate Q) (252 mg, 1.269 mmol) in
THF (20 mL). The mixture was stirred at room temperature for 45
mins. The reaction was quenched with saturated NH.sub.4Cl(aq) and
extracted with EtOAc (2.times.). The combined organic extracts were
washed with brine, dried over sodium sulfate and evaporated under
vacuum. Purification by chromatography on silica, eluting with
20-70% EtOAc/hexane afforded the title compound.
[1052] LC-MS Rt 1.29 mins; [M+H].sup.+ 429.2 (Method
2minLowpHv03)
Step 3:
6-(3-(4-Chlorothiazol-2-yl)-3-hydroxypropyl)-1,3-dimethyl-5-phenyl-
-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1053] Sodium borohydride (415 mg, 10.96 mmol) was added to a
suspension of
6-(3-(4-chlorothiazol-2-yl)-3-oxopropyl)-1,3-dimethyl-5-phenyl-1H-pyrr-
olo[3,4-d]pyrimidine-2,4(3H,6H)-dione (470 mg, 1.096 mmol) in
methanol (20 mL). The mixture was stirred at room temperature for
15 mins. The reaction was quenched with saturated NH.sub.4Cl(aq)
and extracted with chloroform (3.times.). The combined organic
extracts were passed through a hydrophobic frit and evaporated
under vacuum to afford the title compound.
[1054] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.56-7.43 (5H, mult),
7.06 (1H, s), (1H, br s), 4.81 (1H, mult), 4.24 (1H, mult), 4.14
(1H, mult), 3.44 (3H, s), 3.36 (3H, s), 2.35 (1H, mult), 2.14 (1H,
mult).
[1055] LC-MS Rt 1.14 mins [M+H].sup.+ 431.1 (Method
2minLowpHv03)
Step 4:
9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9-dihydro-1H-pyri-
mido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1056] Trifluoromethanesulfonic anhydride (0.576 mL, 3.41 mmol) in
DCM (3 mL) was added slowly to a solution of
6-(3-(4-chlorothiazol-2-yl)-3-hydroxypropyl)-1,3-dimethyl-5-phenyl-1H-pyr-
rolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (490 mg, 1.137 mmol) and
triethylamine (0.792 mL, 5.69 mmol) in DCM (40 mL) at 0.degree. C.
Further portions of triethylamine (0.792 mL, 5.69 mmol) and
trifluoromethanesulfonic anhydride (0.576 mL, 3.41 mmol) were added
to allow the reaction to run to completion. The reaction was
quenched with saturated NaHCO.sub.3(aq) and extracted with DCM. The
organic phase was passed through a hydrophobic frit and evaporated
under vacuum. Purification by mass-directed HPLC under the
following conditions afforded the title compound.
[1057] Column: XSelect CSH Prep C18 column, 30.times.100 mm, 5
um.
[1058] Mobile phase: A=0.1% DEA in water, B=0.1% DEA in MeCN
[1059] Gradient:
[1060] 0.0-0.5 min: 30% B 30 mL/min
[1061] 0.5-1.0 min: 30% B 30-50 mL/min
[1062] 1.0-7.2 min: 30-70% B,
[1063] 7.2-7.3 min: 70-98% B,
[1064] 7.3-9.4 min: 98% B
[1065] 9.4-9.5 min: 30% B 50 mL/min
[1066] 1H NMR (400 MHz, CDCl3) .delta. 7.62 (2H, d), 7.49 (2H, t),
7.42 (1H, t), 7.06 (1H, s), 5.02 (1H, dd), 4.21 (1H, ddd), 4.11
(1H, ddd), 3.40 (6H, apparent s), 3.12 (1H, mult), 2.82 (1H,
dddd).
[1067] LC-MS Rt 1.27 mins; [M+H].sup.+ 413.2 (Method
2minLowpHv03)
Example 16a
(R)-9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9-dihydro-1H-pyrimido-
[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(S)-9-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9-dihydro-1H-pyrimido-
[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00112##
[1069] Chiral separation of racemic
9-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9-dihydro-1H-pyrimido[4,-
5-a]pyrrolizine-2,4(3H,7H)-dione (Example 16) by Supercritical
Fluid Chromatography under the following conditions afforded the
title compound;
[1070] Column: Chiralpak AD-H, 250.times.10 mm, 5 um @ 35 deg
C.
[1071] Mobile phase: 50% Methanol+0.1% v/v DEA/50% CO2
[1072] Flow: 10 ml/min
[1073] Detection: UV @ 220 nm
Example 16a
Enantiomer 1 of
9-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9-dihydro-1H-pyrimido[4,-
5-a]pyrrolizine-2,4(3H,7H)-dione
[1074] Rt=3.68 mins
[1075] 1H NMR (400 MHz, CDCl3) .delta. 7.62 (2H, d), 7.48 (2H, t),
7.42 (1H, t), 7.06 (1H, s), 5.02 (1H, d), 4.21 (1H, mult), 4.11
(1H, mult), 3.40 (6H, apparent s), 3.12 (1H, mult), 2.881 (1H,
dd)
[1076] LC-MS Rt 1.30 mins [M+H].sup.+ 413.2 (Method
2minLowpHv03)
[1077] The second enantiomer was isolated at Rt=5.70 mins
[1078] The following examples were prepared analogously to Example
16 and 16a by replacing 2-bromo-4-chlorothiazole (Intermediate Q)
in step 2 with 2-iodo-4-methylthiazole (Intermediate O).
Example 16.1
1,3-Dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyrimido[4,5-
-a]pyrrolizine-2,4(3H,7H)-dione
##STR00113##
[1080] LC-MS Rt 1.19 mins [M+H] 393.2 (Method 2minLowpHv03)
[1081] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.62 (2H, d), 7.56-7.41
(3H, mult), 7.03 (1H, s), 5.45 (1H, mult), 4.17 (2H, mult), 3.40
(3H, s), 3.37 (3H, s), 3.31 (1H, mult), 2.94 (1H, mult), 2.65 (3H,
s)
Example 16.1a
(R)-1,3-Dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyrimido-
[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(S)-1,3-dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyrimido-
[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00114##
[1083] Column: Chiralpak AD 250 mm.times.10 mm.times.5 .mu.m @ 35
deg C.
[1084] Mobile phase: 40% MeOH (containing 0.1% DEA)/60 CO2
[1085] Flow: 10 ml/min
[1086] Detection: UV @ 220 nm
Example 16.1a
Enantiomer of
1,3-dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyrimido[4,-
5-a]pyrrolizine-2,4(3H,7H)-dione
[1087] Rt=4.01 mins
[1088] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.64 (2H, d), 7.49 (2H,
t), 7.43 (1H, t), 6.89 (1H, s), 5.16 (1H, d), 4.23 (1H, mult), 4.12
(1H, mult), 3.40 (3H, s), 3.38 (3H, s), 3.16 (1H, mult), 2.83 (1H,
mult), 2.53 (3H, s)
[1089] LC-MS Rt 1.20 mins [M+H].sup.+ 393.3 m/z (Method
2minLowpHv03)
[1090] The second enantiomer was isolated at Rt=5.52 mins
Example 17
(8R,10R)-10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(4-methy-
lthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazi-
ne-2,4(3H,10H)-dione or
(8R,10S)-10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(4-methy-
lthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazi-
ne-2,4(3H,10H)-dione or
(8S,10R)-10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(4-methy-
lthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazi-
ne-2,4(3H,10H)-dione or
(8S,10S)-10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(4-methy-
lthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazi-
ne-2,4(3H,10H)-dione
##STR00115##
[1091] Step 1: 2-(((4-Methoxybenzyl)oxy)methyl)oxirane
[1092] p-Methoxybenzyl chloride (2.326 g, 14.85 mmol) was added
slowly to a suspension of sodium hydride (60% dispersion in mineral
oil, 0.594 g, 14.85 mmol) in DMF (25 mL) at 0.degree. C. and the
mixture stirred for 25 mins. Glycidol (1 g, 13.50 mmol) was then
added dropwise over 25 mins, the mixture was allowed to warm to
room temperature and stirred for 3 days. The mixture was diluted
with EtOAc (50 ml) and washed with saturated NH.sub.4Cl (aq) (30
ml), saturated NaHCO.sub.3 (aq) (50 ml) and brine (100 ml). The
organic phase was dried over magnesium sulfate and evaporated under
vacuum. Purification by chromatography on silica, eluting with
0-10% EtOAc/hexane, afforded the title compound.
[1093] LC-MS Rt 1.05 min [M+H].sup.+ 195.8 (Method
2minLowpHv03)
Step 2: 2-(((4-Methoxybenzyl)oxy)methyl)thiirane
[1094] Thiourea (0.935 g, 12.28 mmol) was added to a solution of
2-(((4-methoxybenzyl)oxy)methyl) oxirane (1.084 g, 5.58 mmol) in
methanol (30 mL) and the mixture stirred at room temperature for 16
hours, then evaporated under vacuum. Purification by chromatography
on silica, eluting with 0-10% EtOAc/hexane, afforded the title
compound.
[1095] LC-MS Rt 1.29 min [M-H]-209.0 (Method 2minLowpHv03)
Step 3:
(1-Chloro-3-((4-methoxybenzyl)oxy)propan-2-yl)(trityl)sulfane
[1096] 2-(((4-Methoxybenzyl)oxy)methyl)thiirane (496 mg, 2.359
mmol) was added to a solution of trityl chloride (598 mg, 2.144
mmol) in dichloromethane (10 mL). The mixture was stirred at room
temperature for 16 hours, then evaporated under vacuum.
Purification by chromatography on silica, eluting with 0-5%
EtOAc/hexane, gave a crude material which was triturated with
methanol to afford the title compound.
Step 4:
6-(3-((4-Methoxybenzyl)oxy)-2-(tritylthio)propyl)-1,3-dimethyl-5-(-
4-methylthiazol-2-yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1097] Potassium iodide (9.80 mg, 0.059 mmol) was added to a
mixture of
1,3-dimethyl-5-(4-methylthiazol-2-yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,-
6H)-dione (Intermediate Nd) (81 mg, 0.294 mmol),
(1-chloro-3-((4-methoxybenzyl)oxy)propan-2-yl)(trityl)sulfane (158
mg, 0.323 mmol) and cesium carbonate (191 mg, 0.587 mmol) in
dimethylacetamide (2 ml).
[1098] The mixture was stirred at 50.degree. C. for 3 days. The
reaction mixture was partitioned between water (20 ml) and ethyl
acetate (20 ml), the phases were separated and the aqueous phase
extracted with ethyl acetate (10 ml). The combined organic extracts
were washed with brine (20 ml), dried over magnesium sulfate and
evaporated under vacuum. Purification by chromatography on silica,
eluting with 0-50% EtOAc/hexane, afforded the title compound.
[1099] LC-MS Rt 1.82 min [M+H].sup.+ 729.3 (Method
2minLowpHv03)
Step 5:
10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(4-methyl-
thiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
e-2,4(3H,10H)-dione
[1100] Trifluoroacetic acid (0.155 ml, 2.017 mmol) was added to a
suspension of
6-(3-((4-methoxybenzyl)oxy)-2-(tritylthio)propyl)-1,3-dimethyl-5-(4-methy-
lthiazol-2-yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (98 mg,
0.134 mmol), 5-chlorofuran-2-carbaldehyde (19.30 mg, 0.148 mmol)
and bismuth triflate (44.1 mg, 0.067 mmol) in toluene (2 ml) The
resulting mixture was stirred at room temperature for 30 mins. A
further portion of 5-chlorofuran-2-carbaldehyde (26 mg) was added
and the mixture stirred for a further 16 hours. The mixture was
diluted with 1M NaOH(aq) (10 ml) and DCM (20 ml). The organic phase
was separated and washed with saturated NH.sub.4Cl(aq) (20 ml),
passed through a hydrophobic frit and evaporated under vacuum.
Purification by chromatography on silica, eluting with 0-90%
EtOAc/hexane, afforded the title racemic compound.
[1101] LC-MS Rt 1.26 min [M+H].sup.+ 479.4 (Method
2minLowpHv03)
[1102] The racemate was separated by SFC chromatographic resolution
under the following conditions to afford the title compound as a
single enantiomer.
[1103] Column: Chiralcel OJ-H 250.times.10 mm, 5 um @ 35 deg C.
[1104] Mobile phase: 45% Methanol/55% CO2
[1105] Flow: 10 ml/min
[1106] Detection: UV @ 220 nm
Diastereomer 1 of
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(4-methylthiazol-
-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione
[1107] SFC Rt=4.68 mins
[1108] 1H NMR1 (400 MHz, CDCl.sub.3) .delta. 7.20 (s, 1H), 6.26
(dd, 1H), 6.14 (d, 1H), 5.75 (s, 1H), 5.42 (dd, 1H), 4.04 (dd, 1H),
3.74 (m, 1H), 3.70 (s, 3H), 3.49 (dd, 1H), 3.44 (s, 3H), 3.21 (t,
1H), 2.53 (s, 3H).
[1109] LC-MS Rt 1.29 min [M+H].sup.+ 479.1 (Method
2minLowpHv03)
[1110] The following listed examples were prepared in a similar
manner to Example 17, by replacing
1,3-dimethyl-5-(4-methylthiazol-2-yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,-
6H)-dione (step 4)(Intermediate Nd) with the appropriate starting
material (prepared in a similar manner to intermediate Nd using the
appropriate halo compound). The diastereomers were separated by SFC
chromatographic resolution to afford the title compounds.
Example 17.1
(8R,10R)-10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(1-methy-
l-1H-pyrazol-3-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]th-
iazine-2,4(3H,10H)-dione or
(8R,10S)-10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(1-methy-
l-1H-pyrazol-3-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]th-
iazine-2,4(3H,10H)-dione or
(8S,10R)-10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(1-methy-
l-1H-pyrazol-3-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]th-
iazine-2,4(3H,10H)-dione or
(8S,10S)-10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(1-methy-
l-1H-pyrazol-3-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]th-
iazine-2,4(3H,10H)-dione
##STR00116##
[1112] Separation conditions:
[1113] Column: Chiralcel OJ-H 250.times.10 mm, 5 um @ 35 deg C.
[1114] Mobile phase: 40% Methanol+0.1% v/v DEA/60% CO2
[1115] Flow: 10 ml/min
[1116] Detection: UV @ 220 nm
Diastereoisomer 1 of
10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(1-methyl-1H-pyr-
azol-3-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2-
,4(3H,10H)-dione
[1117] Rt=4.28 mins.
[1118] 1H NMR (400 MHz, DMSO-d6) .delta. 7.82 (1H, d), 6.70 (1H,
d), 6.42 (1H, d), 6.20 (1H, d), 6.15 (1H, s), 5.21 (1H, br s), 4.63
(1H, dd), 4.22 (1H, dd), 3.92 (3H, s), 3.48 (3H, s), 3.46-3.40 (2H,
m), 3.22-3.16 (4H, m).
[1119] LC-MS Rt 1.17 mins [M+H].sup.+ 462.1 (Method
2minLowpHv03)
Example 17.2
(8R,10R)-10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(2-methy-
lthiazol-4-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazi-
ne-2,4(3H,10H)-dione or
(8R,10S)-10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(2-methy-
lthiazol-4-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazi-
ne-2,4(3H,10H)-dione or
(8S,10R)-10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(2-methy-
lthiazol-4-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazi-
ne-2,4(3H,10H)-dione or
(8S,10S)-10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(2-methy-
lthiazol-4-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazi-
ne-2,4(3H,10H)-dione
##STR00117##
[1121] Separation conditions:
[1122] Column: Chiralpak AD-H 250.times.10 mm, 5 um @ 35 deg C.
[1123] Mobile phase: 40% Isopropanol+0.1% v/v DEA/60% CO2
[1124] Flow: 10 ml/min
[1125] Detection: UV @ 220 nm
Diastereoisomer 1 of
10-(5-Chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(2-methylthiazol-
-4-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3-
H,10H)-dione
[1126] Rt=4.69 mins
[1127] 1H NMR (400 MHz, CDCl3) .delta. 7.96 (s, 1H), 6.21 (dd, 1H),
6.11 (d, 1H), 5.72 (s, 1H), 5.17 (dd, 1H), 4.08 (dd, 1H), 3.68 (s,
3H), 3.64 (dd, 1H), 3.46 (dd, 1H), 3.40 (s, 3H), 3.22 (t, 1H), 2.78
(s, 3H).
[1128] LC-MS Rt 1.25 min [M+H].sup.+ 479.0 (Method
2minLowpHv03)
Example 18
9,9-Difluoro-5-(3-fluorophenyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-7,8,9-
,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00118##
[1129] Step 1:
9,9-Difluoro-5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydropyrimido[4,5-a]in-
dolizine-2,4,10(1H,3H,7H)-trione
[1130] A solution of commercially available
N-fluorobenzenesulfonimide (2.77 g, 8.8 mmol) in THF (7 mL) was
added to a suspension of manganese(II) bromide (1.89 g, 8.8 mmol)
in THF (5 mL) and cooled to -78.degree. C. A suspension of
5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydropyrimido[4,5-a]indolizine-2,4,-
10(1H,3H,7H)-trione (Intermediate L, step 1) (750 mg, 2.2 mmol) in
THF (10 mL) was added dropwise, followed by potassium
hexamethyldisilazide solution (1M, 11 mL, 11 mmol). The mixture was
stirred at -60.degree. C. for 2 hours and at room temperature for 1
hour. The reaction was quenched with saturated NaHCO.sub.3(aq) (150
mL) and extracted with DCM (3.times.50 mL). The combined organic
extracts were dried over magnesium sulfate and evaporated under
vacuum. Purification by mass-directed HPLC using the following
conditions afforded the title compound.
[1131] Column: XSelect CSH Prep C18 column, 30.times.100 mm, 5
um.
[1132] Mobile phase: A=0.1% formic acid in water, B=0.1% formic
acid in MeCN
[1133] Elution Gradient:
[1134] 0.0-0.5 min: 30% B 30 mL/min
[1135] 0.5-1.0 min: 30% B 30-50 mL/min
[1136] 1.0-7.2 min: 30-70% B, 7.2-7.3 min: 70-98% B, 7.3-9.4 min:
98% B
[1137] 9.4-9.5 30% B 50 mL/min
[1138] 1H NMR (400 MHz, CDCl3) .delta. 7.55 (1H, mult), 7.31-7.16
(3H, mult), 4.21 (2H, t), 3.95 (3H, s), 3.37 (3H, s), 2.67 (2H,
mult).
[1139] 19F NMR (376 MHz, CDCl3) .delta. -108.7, -110.9.
[1140] LC-MS Rt 1.19 mins [M+H].sup.+ 378.1 (Method
2minLowpHv03)
Step 2:
9,9-Difluoro-5-(3-fluorophenyl)-10-hydroxy-1,3-dimethyl-7,8,9,10-t-
etrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1141] Sodium borohydride (7.2 mg, 0.19 mmol) was added to solution
of
9,9-difluoro-5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydropyrimido[4,5-a]in-
dolizine-2,4,10(1H,3H,7H)-trione (55 mg, 0.15 mmol) in methanol (1
mL) and tetrahydrofuran (1 mL) at 0.degree. C. The mixture was
stirred at room temperature for 30 mins. The reaction was quenched
with water (10 mL) and extracted with DCM (6.times.5 mL). The
combined organic extracts were passed through a hydrophobic frit
and evaporated under vacuum to afford the title compound.
[1142] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.47 (1H, mult),
7.24-7.12 (3H, mult), 5.29 (1H, t), 4.20-4.13 (1H, mult), 3.97 (1H,
mult), 3.77 (3H, s), 3.36 (3H, s), 2.90-2.69 (2H, mult), 2.34 (1H,
mult).
[1143] LC-MS Rt 1.10 mins [M+H].sup.+ 380.1 (Method
2minLowpHv03)
Step 3:
9,9-Difluoro-5-(3-fluorophenyl)-1,3-dimethyl-10-(5-methylfuran-2-y-
l)-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1144] Gold (III) chloride (4 mg, 0.012 mmol) was added to a
suspension of
9,9-difluoro-5-(3-fluorophenyl)-10-hydroxy-1,3-dimethyl-7,8,9,10-tetrahyd-
ropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione (47 mg, 0.12 mmol) and
2-methylfuran (15 .mu.L, 0.16 mmol) in acetonitrile (1 mL). The
mixture was stirred at room temperature for 1 hour. The mixture was
evaporated under vacuum, the residue was dissolved in DCM (5 mL)
and washed with water (5 mL). The organic phase was passed through
a hydrophobic frit evaporated under vacuum. Purification by mass
directed HPLC using the following conditions afforded the title
compound.
[1145] Column: XSelect CSH Prep C18 column, 30.times.100 mm, 5
um.
[1146] Mobile phase: A=0.1% formic acid in water, B=0.1% formic
acid in MeCN
[1147] Elution Gradient:
[1148] 0.0-0.5 min: 40% B 30 mL/min
[1149] 0.5-1.0 min: 40% B 30-50 mL/min
[1150] 1.0-7.2 min: 40-80% B, 7.2-7.3 min: 80-98% B, 7.3-9.4 min:
98% B
[1151] 9.4-9.5 40% B 50 mL/min
[1152] 1H NMR (400 MHz, CDCl3) .delta. 7.48 (1H, mult), 7.26 (1H,
d), 7.22-7.16 (2H, mult), 5.94 (1H, d), 5.90 (1H, d), 5.04 (1H, t),
4.23 (1H, mult), 3.99 (1H, mult), 3.42 (3H, s), 3.34 (3H, s), 2.60
(1H, mult), 2.33-2.23 (4H, mult).
[1153] 19F NMR (376 MHz, CDCl3) .delta. -99.9, -112.3.
[1154] LC-MS Rt 1.39 mins [M+H].sup.+ 444.2 (Method
2minLowpHv03)
Example 19
(8R,10R)-8-(Aminomethyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylt-
hiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-
-2,4(3H,10H)-dione or
(8R,10S)-8-(aminomethyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylt-
hiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-
-2,4(3H,10H)-dione or
(8S,10R)-8-(aminomethyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylt-
hiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-
-2,4(3H,10H)-dione or
(8S,10S)-8-(aminomethyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylt-
hiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-
-2,4(3H,10H)-dione
##STR00119##
[1155] Step 1: 2-(Thiiran-2-ylmethyl)isoindoline-1,3-dione
[1156] Thiourea (1.873 g, 24.61 mmol) was added to a solution of
N-(2,3-epoxypropyl)phthalimide (2 g, 9.84 mmol) in methanol (25
mL). The mixture was stirred at room temperature for 16 hours, then
concentrated under vacuum. The residue was diluted with water (50
ml) and extracted with ethyl acetate (50 ml). The organic extract
was washed with brine (50 ml) passed through a hydrophobic frit and
evaporated under vacuum. Purification by chromatography on silica,
eluting with 0-50% EtOAc/hexane, afforded the title compound.
[1157] LC-MS Rt 1.13 min [M+H].sup.+ 220.4 (Method
2minLowpHv03)
Step 2: 2-(3-Bromo-2-(tritylthio)propyl)isoindoline-1,3-dione
[1158] Trityl bromide (1147 mg, 3.55 mmol) was added portionwise
over 5 mins to a solution of
2-(thiiran-2-ylmethyl)isoindoline-1,3-dione (778 mg, 3.55 mmol) in
DCM (15 mL). The mixture was stirred for at room temperature for 16
hours, diluted with DCM (30 ml) and washed with brine (50 ml). The
organic layer was separated, passed through a hydrophobic frit and
evaporated under vacuum to afford the title compound.
Step 3:
6-(3-(1,3-Dioxoisoindolin-2-yl)-2-(tritylthio)propyl)-1,3-dimethyl-
-5-(4-methylthiazol-2-yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1159] Cesium carbonate (1.537 g, 4.72 mmol) was added to a
solution of
1,3-dimethyl-5-(4-methylthiazol-2-yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,-
6H)-dione (Intermediate Nd) (0.652 g, 2.359 mmol) and
2-(3-bromo-2-(tritylthio)propyl) isoindoline-1,3-dione (2.011 g,
2.59 mmol) in dimethylacetamide (10 ml) and the mixture was stirred
at 55.degree. C. for 16 hours. The mixture was cooled to room
temperature, partitioned between water (150 ml) and ethyl acetate
(150 ml). The organic layer was separated, washed with brine (150
ml), dried over magnesium sulfate and evaporated under vacuum.
Purification by chromatography on silica, eluting with 0-75%
EtOAc/hexane afforded the title compound.
[1160] LC-MS Rt 1.71 min [M+H].sup.+ 738.2 (Method
2minLowpHv03)
Step 4:
10-(5-Chlorofuran-2-yl)-8-((1,3-dioxoisoindolin-2-yl)methyl)-1,3-d-
imethyl-5-(4-methylthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo-
[2,1-c][1,4]thiazine-2,4(3H,10H)-dione
[1161] Trifluoroacetic acid (0.399 ml, 5.18 mmol) was added to a
solution of
6-(3-(1,3-dioxoisoindolin-2-yl)-2-(tritylthio)propyl)-1,3-dimethyl-5-(-
4-methylthiazol-2-yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
(764 mg, 1.035 mmol) in toluene (15 ml) and the mixture was stirred
at room temperature for 30 minutes. Bismuth triflate (340 mg, 0.518
mmol) and 5-chlorofuran-2-carbaldehyde (203 mg, 1.553 mmol) were
then added and the mixture was stirred at room temperature for 16
hours. The mixture was then diluted with ethyl acetate (150 ml) and
washed with water (150 ml), saturated NaHCO.sub.3(aq) (150 ml) and
brine (150 ml). The organic layer was dried over magnesium sulfate
and evaporated under vacuum. Purification by chromatography on
silica, eluting with 0-100% EtOAc/hexane, afforded the title
compound as a mixture of diastereomers.
[1162] LC-MS Rt 1.48 min [M+H].sup.+ 608.6 (Method
2minLowpHv03)
Step 5
8-(aminomethyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2--
yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,1-
0H)-dione as a mixture of stereoisomers
[1163] Ethanolamine (0.320 mL, 5.30 mmol) was added to a solution
of
10-(5-chlorofuran-2-yl)-8-((1,3-dioxoisoindolin-2-yl)methyl)-1,3-dimethyl-
-5-(4-methylthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c]-
[1,4]thiazine-2,4(3H,10H)-dione (537 mg, 0.883 mmol) in toluene (10
mL). The mixture was stirred at 70.degree. C. for 3 hours and then
cooled to room temperature for 16, then heated at 70.degree. C. for
a further 2.5 hours and again cooled to room temperature. The
mixture was diluted with water (100 ml) and ethyl acetate (100 ml).
The organic phase was separated, washed with 1M NaOH(aq) (50 ml)
passed through a hydrophobic frit and evaporated under vacuum to
afford the title compound as a mixture of diastereomers.
[1164] LC-MS Rt 0.82 min [M+H]+ 478.3 and Rt 0.89 min [M+H]+ 478.2
(Method 2minLowpHv03)
[1165] The mixture was separated by SFC chromatographic resolution
under the following conditions to afford the title compound as a
single enantiomer
[1166] Separation conditions:
[1167] Column: Chiralcel OJ-H 250.times.10 mm, 5 um @ 35 deg C.
[1168] Mobile phase: 40% Methanol+0.1% v/v DEA/60% CO2
[1169] Flow: 10 ml/min
[1170] Detection: UV @ 220 nm
Diastereoisomer 1
8-(aminomethyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-
-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,-
10H)-dione
[1171] Rt=3.23 mins LC-MS Rt 0.81 min [M+H].sup.+ 478.1 (Method
2minLowpHv03)
[1172] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.15 (s, 1H), 6.34 (dd,
1H), 6.17 (t, 2H), 4.63 (dd, 1H), 3.79 (dd, 1H), 3.69 (s, 3H), 3.61
(m, 1H), 3.41 (s, 3H), 2.69 (d, 2H), 2.53 (s, 3H)
Example 20
3-(9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9-hexahydro-1-
H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
##STR00120##
[1174] Step 1: Methyl
3-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)propanoate DBU (0.656 mL, 4.35 mmol) was added
to a solution of
3-(1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-
-yl)benzonitrile (Example 9 Step 4) (1.22 g, 4.35 mmol) and methyl
acrylate (0.392 mL, 4.35 mmol) in acetonitrile (15 mL). The mixture
was stirred at room temperature for 4 days. Further portions of
methyl acrylate (0.392 mL, 4.35 mmol) and DBU (0.656 mL, 4.35 mmol)
were added as required to allow the reaction to run to completion.
The reaction was quenched with saturated NH.sub.4Cl(aq) and
extracted with DCM (2.times.). The combined organic extracts were
passed through a hydrophobic frit and evaporated under vacuum.
Purification by chromatography on silica, eluting with 20-100%
EtOAc/hexane afforded a crude material which was triturated with
DCM/diethyl ether/hexane to afford the title compound.
[1175] LC-MS Rt 1.03 mins [M+H].sup.+ 367.1 (Method
2minLowpHv03)
[1176] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.80-7.71 (3H, mult),
7.64 (1H, t), 6.51 (1H, s), 4.24 (2H t), 3.70 (3H, s), 3.43 (3H,
s), 3.36 (3H, s), 2.69 (2H, t)
Step 2:
3-(5-(3-Cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo-
[3,4-d]pyrimidin-6(2H)-yl)-N-methoxy-N-methylpropanamide
[1177] Isopropylmagnesium bromide (2.82 mL, 8.19 mmol) was added
slowly to a suspension of methyl
3-(5-(3-fluorophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d-
]pyrimidin-6(2H)-yl)propanoate (1 g, 2.73 mmol) and
N,O-dimethylhydroxylamine hydrochloride (0.319 g, 3.28 mmol) in THF
(30 mL) and the mixture was stirred at room temperature for 30
mins. The reaction was quenched with saturated NH.sub.4Cl(aq) and
extracted with DCM (2.times.). The combined organic extracts were
passed through a hydrophobic frit and evaporated under vacuum.
Trituration with DCM/hexane afforded the title compound.
[1178] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.76 (3H, mult), 7.64
(1H, t), 6.57 (1H, s), 4.27 (2H, t), 3.62 (3H, s), 3.44 (3H, s),
3.36 (3H, s), 3.18 (3H, s), 2.78 (2H, t).
[1179] LC-MS Rt 0.97 mins [M+H].sup.+ 396.5 (Method
2minLowpHv03)
Step 3:
3-(6-(3-(4-Chlorothiazol-2-yl)-3-oxopropyl)-1,3-dimethyl-2,4-dioxo-
-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
[1180] Isopropylmagnesium chloride lithium chloride complex
solution (4.58 mL, 5.95 mmol) was added slowly to a suspension of
3-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)-N-methoxy-N-methylpropanamide (784 mg, 1.983
mmol) and 2-bromo-4-chlorothiazole (Intermediate Q, step 2) (394
mg, 1.983 mmol) in THF (30 mL). The mixture was stirred at room
temperature for 40 mins. The reaction mixture was quenched with
saturated NH.sub.4Cl(aq) and extracted with EtOAc (2.times.). The
combined organic extracts were washed with brine, dried over sodium
sulphate and evaporated under vacuum. Purification by
chromatography on silica, eluting with 10-60% EtOAc/hexane afforded
the title compound.
[1181] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.79-7.59 (4H, mult),
7.51 (1H, s), 6.51 (1H, s), 4.41 (2H, t), 3.50 (2H, t), 3.41 (3H,
s), 3.35 (3H, s)
[1182] LC-MS Rt 1.20 mins [M+H].sup.+ 454.0 (Method
2minLowpHv03)
Step 4:
3-(6-(3-(4-Chlorothiazol-2-yl)-3-hydroxypropyl)-1,3-dimethyl-2,4-d-
ioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
[1183] Sodium borohydride (408 mg, 10.80 mmol) was added to a
solution of
3-(6-(3-(4-chlorothiazol-2-yl)-3-oxopropyl)-1,3-dimethyl-2,4-dioxo-2,3,4,-
6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile (490 mg,
1.080 mmol) in methanol (20 mL). The mixture was stirred at room
temp. for 25 mins, then quenched with saturated NaHCO.sub.3(aq) and
extracted with chloroform (3.times.). The combined organic extracts
were passed through a hydrophobic frit and evaporated under vacuum
to afford the title compound.
[1184] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.80-7.59 (4H, mult),
7.09 (1H, s), 6.51 (1H, s), 4.91 (1H, mult), 4.16 (2H, mult), 3.43
(3H, s), 3.37 (3H, s), 2.33 (1H, mult), 2.24 (1H, mult).
[1185] LC-MS Rt 1.11 mins [M+H].sup.+ 456.1 (Method
2minLowpHv03)
Step 5:
3-(9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9-hex-
ahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
[1186] Trifluoromethanesulfonic anhydride (0.445 mL, 2.63 mmol) was
added to a solution of
3-(6-(3-(4-chlorothiazol-2-yl)-3-hydroxypropyl)-1,3-dimethyl-2,4-dioxo-2,-
3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile (600
mg, 1.316 mmol) and triethylamine (0.550 mL, 3.95 mmol) in DCM (20
mL). The mixture was stirred at room temperature for 90 mins.
Further portions of triethylamine (0.550 mL, 3.95 mmol) and
trifluoromethanesulfonic anhydride (0.445 mL, 2.63 mmol) were added
to allow the reaction to run to completion. The reaction mixture
was quenched with saturated NaHCO.sub.3(aq) and extracted with DCM
(3.times.). The combined organic extracts were passed through a
hydrophobic frit and evaporated under vacuum. Purification by
chromatography on silica, eluting with 20-80% EtOAc/hexane gave a
crude material which was triturated with DCM/diethyl ether/hexane
to afford the title compound.
[1187] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.92 (1H, mult), 7.88
(1H, mult), 7.71 (1H, mult), 7.61 (1H, mult), 7.08 (1H, s), 5.04
(1H, dd), 4.26 (1H, mult), 4.11 (1H, mult), 3.41 (3H, s), 3.40 (3H,
s), 3.15 (1H, mult), 2.86 (1H, mult).
[1188] LC-MS Rt 1.23 mins [M+H].sup.+ 438.1 (Method
2minLowpHv03)
Example 20a
(R)-3-(9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9-hexahyd-
ro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile or
(S)-3-(9-(4-chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9-hexahyd-
ro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
##STR00121##
[1190] Chiral separation of the racemic Example 20 by Supercritical
Fluid Chromatography using the following conditions afforded the
title compound:
[1191] Column: Chiralpak IA, 250.times.10 mm, 5 um @ 35.degree.
C.,
[1192] Mobile phase: 50% Methanol/50% CO2
[1193] Flow: 10 ml/min
[1194] Detection: UV @ 220 nm
Example 20a
Enantiomer 1 of
3-(9-(4-chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9-hexahydro--
1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile SFC
[1195] Rt=4.77 mins,
[1196] 1H NMR (400 MHz, CDCl3) .delta. 7.93 (1H, mult), 7.88 (1H,
s), 7.71 (1H, mult), 7.61 (1H, t), 7.08 (1H, s), 5.04 (1H, d), 4.26
(1H, mult), 4.11 (1H, mult), 3.41 (3H, s), 3.41 (3H, s, distinct
peak), 3.16 (1H, mult), 2.86 (1H, mult)
[1197] LC-MS Rt 1.24 mins [M+H].sup.+ 438.0 m/z (Method
2minLowpHv03)
[1198] The other enantiomer was isolated at Rt=6.97 mins
[1199] The following compounds were prepared analogously to
Examples 20 by replacing
3-(1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]p-
yrimidin-5-yl)benzonitrile with the appropriate starting compound
(described hereinafter). Example 20.2 was separated by SFC
chromatography to afford Example 20.2a
Example 20.1
9-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8,9-dihydro-
-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00122##
[1201] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.04 (1H, s), 7.02 (1H,
s), 5.04 (1H, mult), 4.97 (1H, mult), 4.52 (1H, mult), 3.47 (3H,
s), 3.40 (3H, s), 3.18 (1H, mult), 2.83 (1H, mult), 2.51 (3H,
s)
[1202] LC-MS Rt 1.39 mins [M+H].sup.+ 434.4 (Method
2minLowpHv03)
Example 20.2
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydro-1H-py-
rimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00123##
[1204] LC-MS Rt 1.30 mins [M+H].sup.+ MS 431.5 (Method
2minLowpHv03)
[1205] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.49-7.39 (2H, mult),
7.34 (1H, mult), 7.12 (1H, mult), 7.06 (1H, s), 5.02 (1H, dd), 4.21
(1H, mult), 4.11 (1H, mult), 3.40 (6H, apparent s), 3.12 (1H,
mult), 2.83 (1H, mult).
Example 20.2a
(R)-9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydro-1-
H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(S)-9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydro-1-
H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00124##
[1207] Separation conditions:
[1208] Column: Chiralpak AD-H, 250.times.10 mm, 5 um @ 35 deg
C.
[1209] Mobile phase: 50% Methanol/50% CO2
[1210] Flow: 10 ml/min
[1211] Detection: UV @ 220 nm
Example 20.2a
Enantiomer 1 of
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydro-1H-p-
yrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1212] Rt=3.24 mins
[1213] 1H NMR (400 MHz, CDCl3) .delta. 7.49-7.37 (2H, mult), 7.34
(1H, d), 7.11 (1H, t), 5.02 (1H, d), 4.22 (1H, mult), 4.11 (1H, t),
3.39 (3H, s), 3.39 (3H, s, distinct peak), 3.12 (1H, mult), 2.82
(1H, dd)
[1214] LC-MS Rt 1.30 mins [M+H].sup.+ 431.1 (Method
2minLowpHv03)
Example 21
9-(4-Chlorothiazol-2-yl)-1,3,8,8-tetramethyl-5-phenyl-8,9-dihydro-1H-pyrim-
ido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00125##
[1215] Step 1:
7-((4-Chlorothiazol-2-yl)(hydroxy)methyl)-1,3-dimethyl-5-phenyl-1H-pyrrol-
o[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1216] Isopropylmagnesium chloride lithium chloride complex
solution (1.3M, 2.82 mL, 3.67 mmol) was added slowly to
2-bromo-4-chlorothiazole (Intermediate Q, step 2) (729 mg, 3.67
mmol) in THF (2 mL) at 0.degree. C. The mixture was stirred at
0.degree. C. for 5 mins, then added dropwise to
1,3-dimethyl-2,4-dioxo-5-phenyl-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrim-
idine-7-carbaldehyde (Intermediate Na) (416 mg, 1.469 mmol) in THF
(8 mL) at 0.degree. C. and the resultant mixture stirred for 15
mins. The reaction mixture was quenched with saturated
NH.sub.4Cl(aq) and extracted with DCM (3.times.). The combined
organic extracts were evaporated under vacuum. Purification by
chromatography on silica, eluting with 40-100% EtOAc/hexane then
5-10% MeOH/EtOAc afforded a crude material which was triturated
with DCM/EtOAc/diethyl ether/hexane to afford the title
compound.
[1217] LC-MS Rt 1.11 mins; [M+H].sup.+ 403.1 (Method
2minLowpHv03)
[1218] 1H NMR (400 MHz, DMSO-d6) .delta. 12.04 (1H, s), 7.79-7.73
(3H, mult), 7.48-7.38 (3H, mult), 7.25 (1H, d), 6.39 (1H, d), 3.47
(3H, s), 3.22 (3H, s).
Step 2: Methyl
3-(4-chlorothiazol-2-yl)-3-(1,3-dimethyl-2,4-dioxo-5-phenyl-2,3,4,6-tetra-
hydro-1H-pyrrolo[3,4-d]pyrimidin-7-yl)-2,2-dimethylpropanoate
[1219] Boron trifluoride THF complex (0.028 mL, 0.255 mmol) was
added to a solution of
7-((4-chlorothiazol-2-yl)(hydroxy)methyl)-1,3-dimethyl-5-phenyl-1H-pyrrol-
o[3,4-d]pyrimidine-2,4(3H,6H)-dione (114 mg, 0.283 mmol) and
commercially available
((1-methoxy-2-methylprop-1-en-1-yl)oxy)trimethylsilane (0.143 mL,
0.707 mmol) in THF (5 mL). The mixture was stirred at room
temperature for 5 minutes. The reaction was quenched with saturated
NaHCO.sub.3(aq) and extracted with DCM (2.times.). The combined
organic extracts were passed through a hydrophobic frit and
evaporated under vacuum. Purification by chromatography on silica,
eluting with 20-80% EtOAc/hexane afforded the title compound.
[1220] 1H NMR (400 MHz, DMSO-d6) .delta. 10.83 (1H, s), 7.92 (2H,
d), 7.51 (2H, t), 7.43 (1H, t), 7.04 (1H, s), 5.11 (1H, s), 3.75
(3H, s), 3.73 (3H, s), 3.44 (3H, s), 1.53 (3H, s), 1.33 (3H,
s).
[1221] LC-MS Rt 1.49 mins [M+H].sup.+ 487.5 (Method
2minLowpHv03)
Step 3:
7-(1-(4-Chlorothiazol-2-yl)-3-hydroxy-2,2-dimethylpropyl)-1,3-dime-
thyl-5-phenyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1222] Sodium borohydride (46.6 mg, 1.232 mmol) was added to a
mixture of methyl 3-(4-chloro
thiazol-2-yl)-3-(1,3-dimethyl-2,4-dioxo-5-phenyl-2,3,4,6-tetrahydro-1H-py-
rrolo[3,4-d]pyrimidin-7-yl)-2,2-dimethylpropanoate (120 mg, 0.246
mmol) and lithium chloride (52.2 mg, 1.232 mmol) in methanol (10
mL). The mixture was stirred at room temp. for 30 mins. The
reaction was quenched with saturated NaHCO.sub.3(aq) and extracted
with DCM (3.times.). The combined organic extracts were passed
through a hydrophobic frit and evaporated under vacuum to afford
the title compound.
[1223] 1H NMR (400 MHz, CDCl.sub.3) .delta. 10.71 (1H, s), 7.89
(2H, d), 7.50 (2H, t), 7.41 (1H, t), 7.04 (1H, s), 5.05 (1H, s),
3.73 (3H, s), 3.50 (1H, d), 3.43 (3H, s), 3.38 (1H, d), 1.19 (3H,
s), 1.10 (3H, s).
[1224] LC-MS Rt 1.37 mins [M+H].sup.+ 459.1 (Method
2minLowpHv03)
Step 4:
3-(4-Chlorothiazol-2-yl)-3-(1,3-dimethyl-2,4-dioxo-5-phenyl-2,3,4,-
6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-7-yl)-2,2-dimethylpropyl
methanesulfonate
[1225] Methanesulfonyl chloride (0.034 mL, 0.436 mmol) was added to
a solution of
7-(1-(4-chlorothiazol-2-yl)-3-hydroxy-2,2-dimethylpropyl)-1,3-dimethyl-5--
phenyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (100 mg, 0.218
mmol) and triethylamine (0.076 mL, 0.545 mmol) in DCM (6 mL). The
mixture was stirred at room temperature for 5 mins. The reaction
was quenched with saturated NaHCO.sub.3(aq) and extracted with DCM.
The organic phase was passed through a hydrophobic frit and
evaporated under vacuum to afford the title compound as a crude
material which was used without further purification;
[1226] LC-MS Rt 1.40 mins [M+H].sup.+ 537.5 (Method
2minLowpHv03)
Step 5:
9-(4-Chlorothiazol-2-yl)-1,3,8,8-tetramethyl-5-phenyl-8,9-dihydro--
1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1227] Sodium hydride (60% wt., 38.0 mg, 0.950 mmol) was added to a
solution of
3-(4-chlorothiazol-2-yl)-3-(1,3-dimethyl-2,4-dioxo-5-phenyl-2,3,4,6-tetra-
hydro-1H-pyrrolo[3,4-d]pyrimidin-7-yl)-2,2-dimethylpropyl
methanesulfonate (170 mg, 0.317 mmol) in THF (10 mL). The mixture
was stirred at room temperature for 5 mins. The reaction was
quenched with water and extracted with DCM (2.times.). The combined
organic extracts were passed through a hydrophobic frit and
evaporated under vacuum. Purification by chromatography on silica,
eluting with 20-100% EtOAc/hexane afforded the title compound.
[1228] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.66 (2H, d), 7.50 (2H,
t), 7.44 (1H, t), 7.09 (1H, s), 4.55 (1H, s), 4.10 (1H, d), 3.78
(1H, d), 3.40 (3H, s), 3.33 (3H, s), 1.37 (3H, s), 1.06 (3H, s)
[1229] LC-MS Rt 1.40 mins [M+H].sup.+ 441.1 (Method
2minLowpHv03)
Example 21a
(S)-9-(4-Chlorothiazol-2-yl)-1,3,8,8-tetramethyl-5-phenyl-8,9-dihydro-1H-p-
yrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(R)-9-(4-chlorothiazol-2-yl)-1,3,8,8-tetramethyl-5-phenyl-8,9-dihydro-1H-p-
yrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00126##
[1231] Chiral separation of the racemic Example 21 by Supercritical
Fluid Chromatography was carried out using the following conditions
to afford the title compound:
[1232] Column: Chiralcel OJ-H 250.times.10 mm, 5 um @ 35 deg C.
[1233] Mobile phase: 30% Methanol/70% CO2
[1234] Flow: 10 ml/min
[1235] Detection: UV @ 220 nm
Example 21a
Enantiomer 1 of
9-(4-chlorothiazol-2-yl)-1,3,8,8-tetramethyl-5-phenyl-8,9-dihydro-1H-pyri-
mido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1236] Rt=5.48 mins
[1237] 1H NMR (400 MHz, CDCl3) .delta. 7.55 (2H, d), 7.40 (2H, t),
7.33 (1H, t), 7.00 (1H, s), 4.45 (1H, s), 4.00 (1H, d), 3.68 (1H,
d), 3.30 (3H, s), 3.23 (3H, s), 1.27 (3H, s), 0.96 (3H, s)
[1238] LC-MS Rt 1.39 mins [M+H].sup.+ 441.1 (Method
2minLowpHv03)
[1239] The other enantiomer was isolated at Rt=4.27 mins
[1240] The following listed compounds were prepared in a similar
manner to Example 21, using the appropriate starting material (the
preparations of which are described hereinafter):
Example 21.1
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3,8,8-tetramethyl-8,9-dihydr-
o-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00127##
[1242] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.50-7.43 (2H, mult),
7.38 (1H, mult), 7.15 (1H, mult), 7.10 (1H, s), 4.55 (1H, s), 4.11
(1H, d), 3.77 (1H, d), 3.40 (3H, s), 3.33 (3H, s), 1.37 (3H, s),
1.07 (3H, s).
[1243] LC-MS Rt 1.42 mins [M+H].sup.+ 459.4 (Method
2minLowpHv03)
Example 21.2
9-(4-Chlorothiazol-2-yl)-1,3,8,8-tetramethyl-5-phenyl-8,9-dihydro-1H-pyrim-
ido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00128##
[1245] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.66 (2H, d), 7.50 (2H,
t), 7.44 (1H, t), 7.09 (1H, s), 4.55 (1H, s), 4.10 (1H, d), 3.78
(1H, d), 3.40 (3H, s), 3.33 (3H, s), 1.37 (3H, s), 1.06 (3H, s)
[1246] LC-MS Rt 1.40 mins [M+H].sup.+ 441.1 (Method
2minLowpHv03)
Example 21.3
3-(1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-2,4-dioxo-2,3,4,7,8,9-hexa-
hydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
##STR00129##
[1248] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.99 (1H, d), 7.88 (1H,
s), 7.71 (1H, d), 7.62 (1H, t), 6.99 (1H, s), 4.88 (1H, s), 4.16
(1H, d), 3.76 (1H, d), 3.40 (3H, s), 3.33 (3H, s), 2.59 (3H, s),
1.43 (3H, s), 1.11 (3H, s)
[1249] LC-MS Rt 1.30 mins; [M+H].sup.+ 446.4 (Method
2minLowpHv03)
Example 21.4
5-(3-Fluorophenyl)-1,3,8,8-tetramethyl-9-(4-methylthiazol-2-yl)-8,9-dihydr-
o-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00130##
[1251] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.50-7.43 (2H, mult),
7.37 (1H, d), 7.14 (1H, mult), 7.00 (1H, s), 4.95 (1H, s), 4.10
(1H, d), 3.79 (1H, d), 3.40 (3H, s), 3.32 (3H, s), 2.61 (3H, s),
1.44 (3H, s), 1.12 (3H, s).
[1252] LC-MS Rt 1.35 mins [M+H].sup.+ 439.3 (Method
2minLowpHv03)
Example 21.5
1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyrim-
ido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00131##
[1254] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.65 (2H, d), 7.51 (2H,
t), 7.45 (1H, t), 7.03 (1H, s), 5.03 (1H, s), 4.08 (1H, d), 3.81
(1H, d), 3.40 (3H, s), 3.32 (3H, s), 2.64 (3H, s), 1.45 (3H, s),
1.13 (3H, s)
[1255] LC-MS Rt 1.34 mins [M+H].sup.+ 421.5 (Method
2minLowpHv03)
[1256] The compounds of the following listed Examples were prepared
by SFC chromatographic resolution of the appropriate racemate.
Example 21.6
(R)-9-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3,8,8-tetramethyl-8,9-di-
hydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(S)-9-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3,8,8-tetramethyl-8,9-di-
hydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00132##
[1258] Separation conditions:
[1259] Column: Chiralpak AD-H 250.times.10 mm, 5 um @ 35 deg C.
[1260] Mobile phase: 50% Methanol/50% CO2
[1261] Flow: 10 ml/min
[1262] Detection: UV @ 220 nm
Enantiomer 1 of
9-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3,8,8-tetramethyl-8,9-dihyd-
ro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1263] Rt=1.99 mins
[1264] 1H NMR (400 MHz, CDCl3) .delta. 7.50-7.42 (2H, mult), 7.37
(1H, mult), 7.13 (1H, mult), 7.09 (1H, s), 4.54 (1H, s), 4.11 (1H,
d), 3.77 (1H, d), 3.39 (3H, s), 3.32 (3H, s), 1.36 (3H, s), 1.06
(3H, s)
[1265] LC-MS Rt 1.45 mins [M+H].sup.+ 459.3 (Method
2minLowpHv03)
Example 21.7
(R)-5-(3-Fluorophenyl)-1,3,8,8-tetramethyl-9-(4-methylthiazol-2-yl)-8,9-di-
hydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(S)-5-(3-Fluorophenyl)-1,3,8,8-tetramethyl-9-(4-methylthiazol-2-yl)-8,9-di-
hydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00133##
[1267] Separation conditions:
[1268] Column: Chiralpak AD-H 250.times.10 mm, 5 um @ 35 deg C.
[1269] Mobile phase: 20% methanol/80% CO2
[1270] Flow: 10 ml/min
[1271] Detection: UV @ 220 nm
Enantiomer 1 of
5-(3-fluorophenyl)-1,3,8,8-tetramethyl-9-(4-methylthiazol-2-yl)-8,9-dihyd-
ro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1272] Rt=3.13 mins
[1273] 1H NMR (400 MHz, CDCl3) .delta. 7.45 (2H, mult), 7.38 (1H,
mult), 7.12 (1H, mult), 6.85 (1H, s), 4.54 (1H, s), 4.14 (1H, d),
3.75 (1H, d), 3.39 (3H, s), 3.31 (3H, s), 2.48 (3H, s), 1.35 (3H,
s), 1.02 (3H, s)
[1274] LC-MS Rt 1.39 mins [M+H].sup.+ 439.3 (Method
2minLowpHv03)
Example 21.8
(R)-3-(1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-2,4-dioxo-2,3,4,7,8,9--
hexahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile or
(S)-3-(1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-2,4-dioxo-2,3,4,7,8,9--
hexahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
##STR00134##
[1276] Separation conditions:
[1277] Column: Chiralpak AD-H, 250.times.10 mm, 5 um @ 35 deg
C.
[1278] Mobile phase: 20% Methanol/80% CO2
[1279] Flow: 10 ml/min
[1280] Detection: UV @ 220 nm
Enantiomer 1 of
3-(1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-2,4-dioxo-2,3,4,7,8,9-hex-
ahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
[1281] Rt=3.80 mins
[1282] 1H NMR (400 MHz, CDCl3) .delta. 7.99 (1H, d), 7.88 (1H, s),
7.72 (1H, d), 7.63 (1H, t), 7.02 (1H, s), 4.97 (1H, s), 4.15 (1H,
d), 3.76 (1H, d), 3.40 (3H, s), 3.33 (3H, s), 2.62 (3H, s), 1.45
(3H, s), 1.13 (3H, s).
[1283] LC-MS Rt 1.30 mins; [M+H].sup.+ 446.4 (Method
2minLowpHv03)
Example 21.9
(R)-1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-p-
yrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(S)-1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-p-
yrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00135##
[1285] Separation conditions:
[1286] Column: Chiralpak AD-H 250.times.10 mm, 5 um @ 35 deg C.
[1287] Mobile phase: 25% Methanol/75% CO2
[1288] Flow: 10 ml/min
[1289] Detection: UV @ 220 nm
Enantiomer 1 of
1,3,8,8-Tetramethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihydro-1H-pyri-
mido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1290] Rt=2.89 mins
[1291] 1H NMR (400 MHz, CDCl3) .delta. 7.65 (2H, d), 7.48 (2H, t),
7.41 (1H, t), 6.83 (1H, s), 4.54 (1H, s), 4.13 (1H, d), 3.74 (1H,
d), 3.38 (3H, s), 3.31 (3H, s), 2.48 (3H, s), 1.35 (3H, s), 1.00
(3H, s)
[1292] LC-MS Rt 1.33 mins; [M+H].sup.+ 421.3 (Method
2minLowpHv03)
Example 22
10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8,9,-
10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00136##
[1293] Step 1:
1,3-Dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4-
(3H,6H)-dione
[1294] The title compound was prepared by a similar method to
3-(1,3-Dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-
-yl)benzonitrile (Example 9.0 Step 4), replacing
3-bromobenzonitrile with 3-iodo-1-methyl-1H-pyrazole in Step 3.
[1295] 1H NMR (400 MHz, DMSO-d6) .delta. 12.07 (1H, s), 7.74 (1H,
d), 7.36 (1H, d), 6.67 (1H, s), 3.91 (3H, s), 3.30 (3H, s), 3.24
(3H, s).
[1296] LC-MS Rt 1.85 mins [M+H].sup.+ 259.0 (Method
5minLowpHv01)
Step 2:
4-(1,3-Dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-2,4-dioxo-3,4-dihydro-
-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)-N-methoxy-N-methylbutanamide
[1297]
1,3-Dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-1H-pyrrolo[3,4-d]pyrimidi-
ne-2,4(3H,6H)-dione (800 mg, 3.09 mmol),
4-bromo-N-methoxy-N-methylbutanamide (Intermediate P) (843 mg, 4.01
mmol) and cesium carbonate (2011 mg, 6.17 mmol) were suspended in
DMF (11.100 ml) and the mixture was stirred at room temperature for
22 hours. The mixture was poured into water (50 ml) and extracted
with DCM (3.times.100 ml). The combined organic extracts were
washed with water (2.times.50 ml) and brine (50 ml), dried over
sodium sulfate and evaporated under vacuum. Purification by
mass-directed HPLC under the following conditions afforded the
title compound.
[1298] Column: XSelect CSH Prep C18 column, 30.times.100 mm, 5
um.
[1299] Mobile phase: A=0.1% DEA in water, B=0.1% DEA in MeCN
[1300] Gradient:
[1301] 0.0-0.5 min: 10% B 30 mL/min
[1302] 0.5-1.0 min: 10% B 30-50 mL/min
[1303] 1.0-7.2 min: 10-35% B, 7.2-7.3 min: 35-98% B, 7.3-9.4 min:
98% B
[1304] 9.4-9.5 10% B 50 mL/min
[1305] 1H NMR (400 MHz, DMSO-d6) .delta. 7.76 (1H, d), 6.91 (1H,
s), 6.84 (1H, d), 4.27 (2H, t), 3.91 (3H, s), 3.59 (3H, s), 3.30
(3H, s), 3.19 (3H, s), 3.05 (3H, s), 2.32 (2H, t), 1.92 (2H,
quint).
[1306] LC-MS Rt 0.87 mins [M+H].sup.+ 389.6 (Method
2minLowpHv03)
Step 3:
6-(4-(4-Chlorothiazol-2-yl)-4-oxobutyl)-1,3-dimethyl-5-(1-methyl-1-
H-pyrazol-3-yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1307] Isopropylmagnesium chloride lithium chloride complex (3M
solution in THF, 1.188 ml, 1.545 mmol) was added slowly to a
solution of
4-(1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-2,4-dioxo-3,4-dihydro-1H-pyr-
rolo[3,4-d]pyrimidin-6(2H)-yl)-N-methoxy-N-methylbutanamide (200
mg, 0.515 mmol) and 2-bromo-4-chlorothiazole (Intermediate Q, step
2) (112 mg, 0.566 mmol) in THF (3.280 ml) at 0.degree. C. The
mixture was then stirred at room temperature for 2 hours. A further
portion of isopropylmagnesium chloride lithium chloride complex (3M
solution in THF, 300 .mu.l) was added and the mixture stirred for a
further 15 mins. The reaction was quenched with saturated
NH.sub.4Cl(aq) (6 ml), diluted with water (20 ml) and extracted
with EtOAc (3.times.30 ml). The combined organic extracts were
washed with brine (10 ml), dried over sodium sulfate and evaporated
under vacuum. Purification by chromatography on silica, eluting
with 50-70% EtOAc/hexane, afforded the title compound.
[1308] 1H NMR (400 MHz, DMSO-d6) .delta. 8.22 (1H, s), 7.70 (1H,
d), 6.93 91H, s), 6.82 (1H, d), 4.35 (2H, t), 3.86 (3H, s), 3.26
(3H, s), 3.19 (3H, s), 306 (2H, t), 2.09 (2H, t)
[1309] LC-MS Rt 1.15 mins [M+H].sup.+ 447.2 (Method
2minLowpHv03)
Step 4:
10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl-
)-7,8-dihydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1310] Pyridine hydrochloride (6.02 g, 52.1 mmol) was added to a
suspension of
6-(4-(4-chlorothiazol-2-yl)-4-oxobutyl)-1,3-dimethyl-5-(1-methyl-1H-pyraz-
ol-3-yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (2.33 g, 5.21
mmol) in methanol (82 ml) and water (8.17 ml) and the mixture
stirred at room temperature for 3 days. The mixture was then
diluted with chloroform (300 ml), washed with 1M HCl(aq)
(3.times.100 ml) and saturated NaHCO.sub.3(aq) (100 ml). The
organic phase was passed through a hydrophobic frit and evaporated
under vacuum. Purification by chromatography on silica, eluting
with 50-80% EtOAc/hexane afforded a crude material which was
triturated with methanol to afford the title compound.
[1311] 1H NMR (400 MHz, DMSO-d6) .delta. 7.83 (1H, d), 7.80 (1H,
s), 6.94 (1H, d), 6.66 (1H, t), 4.15 (2H, t), 3.96 (3H, s), 3.19
(3H, s), 2.68 (3H, s), 2.57 (2H, q).
[1312] LC-MS Rt 1.16 mins [M+H].sup.+ 429.3 (Method
2minLowpHv03)
Step 5:
10-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl-
)-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1313] A suspension of
10-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-(1-methyl-1H-pyrazol-3-yl)-7,8-d-
ihydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione (25 mg, 0.058
mmol) and 10% platinum on carbon (12 mg, 6.15 .mu.mol) in ethanol
(2.2 ml) and THF (6.6 ml) was stirred under an atmosphere of
hydrogen for 7 days. The mixture was filtered and the residue
rinsed thoroughly with ethanol and the combined filtrates
evaporated under vacuum. Purification by chromatography on silica,
eluting with 80% EtOAc/hexane afforded a crude material which was
triturated with diethyl ether to afford the title compound.
[1314] 1H NMR (400 MHz, DMSO-d6) .delta. 7.79 (1H, d), 7.61 (1H,
s), 6.72 (1H, d), 5.28 (1H, m), 4.46 (1H, obs d), 3.39 (3H, s),
3.88 (1H, m), 3.30 (3H, s), 3.19 (3H, s), 2.39-2.31 (2H, m), 2.25
(1H, t), 1.87 (1H, d), 1.64 (1H, q)
[1315] LC-MS Rt 1.14 mins [M+H].sup.+ 431.4 (Method
2minLowpHv03)
Example 23a, 23b and 23c
(8R,10R)-10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,-
3-dimethyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
or
(8R,10S)-10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,-
3-dimethyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
or
(8S,10R)-10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,-
3-dimethyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
or
(8S,10S)-10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,-
3-dimethyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00137## ##STR00138##
[1316] Step 1: 4-(Hydroxymethyl)furan-2(5H)-one
[1317] (Carbethoxymethylene)triphenylphosphorane (63.8 g, 183 mmol)
was added in four portions to a suspension of 1,3-dihydroxyacetone
(15.0 g, 167 mmol) in DCM (165 mL). The mixture was stirred at room
temperature for 18 hours. The reaction mixture was extracted with
water (4.times.150 mL). The combined aqueous phases were treated
with 3 g of decolourising charcoal and filtered, washing the
residue with a little water. The filtrates were evaporated under
vacuum, then evaporated again from toluene to afford the title
compound.
[1318] 1H NMR (400 MHz, CDCl.sub.3) .delta. 6.05 (1H, s), 4.88 (2H,
s), 4.61 (2H, s), 2.46 (1H, br).
Step 2: 4-(Hydroxymethyl)dihydrofuran-2(3H)-one
[1319] A mixture of 4-(hydroxymethyl)furan-2(5H)-one (1.0 g, 8.8
mmol) and 10% palladium on carbon (100 mg) in
2-methyltetrahydrofuran (9 mL) was stirred under an atmosphere of
hydrogen for 23 hours. The reaction mixture was filtered and the
residue rinsed with ethyl acetate. The filtrates were evaporated
under vacuum to afford the title compound.
[1320] 1H NMR (400 MHz, CDCl.sub.3) .delta. 4.14 (1H, dd), 4.24
(1H, dd), 3.74-3.66 (2H, mult), 2.78 (1H, mult), 2.63 (1H, dd),
2.41 (1H, dd), 2.26 (1H, br).
Step 3: (5-Oxotetrahydrofuran-3-yl)methyl
trifluoromethanesulfonate
[1321] A solution of trifluoromethanesulfonic anhydride (15.0 g,
53.2 mmol) in DCM (25 mL) was added dropwise to a solution of
4-(hydroxymethyl)dihydrofuran-2(3H)-one (4.75 g, 41.0 mmol) and
2,6-lutidine (7.15 mL, 61.4 mmol) in DCM (111 mL) at 0.degree. C.
The mixture was stirred at 0.degree. C. for 20 minutes. The product
mixture was diluted with DCM (100 mL) and was washed with water
(3.times.100 mL). The organic phase was dried over magnesium
sulfate and evaporated under vacuum.
[1322] Purification by chromatography on silica, eluting with
30-50% EtOAc/hexane, afforded the title compound.
[1323] 1H NMR (400 MHz, CDCl.sub.3) .delta. 4.60-4.49 (3H, mult),
4.22 (1H, dd), 3.13 (1H, mult), 2.79 (1H, dd), 2.43 (1H, dd).
[1324] 19F NMR (376 MHz, CDCl.sub.3) .delta. -74.2.
Step 4:
5-(3-fluorophenyl)-1,3-dimethyl-6-((5-oxotetrahydrofuran-3-yl)meth-
yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1325]
5-(3-Fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,-
6H)-dione (2.50 g, 9.2 mmol) was added portionwise to a mixture of
potassium tert-butoxide (1.18 g, 10.5 mmol) and 18-crown-6 (242 mg,
0.92 mmol) in THF (15 mL), and stirred at room temperature for 15
minutes. A solution of (5-oxotetrahydrofuran-3-yl)methyl
trifluoromethanesulfonate (2.61 g, 10.5 mmol) in THF (7.5 mL) was
then added, maintaining the temperature at .about.20.degree. C.
using a water bath. The solution was stirred at room temperature
for 1 hour. The reaction was quenched with saturated NH.sub.4Cl(aq)
(10 mL), diluted with dichloromethane (100 mL) and washed with
saturated NH.sub.4Cl(aq) (2.times.50 mL). The organic phase was
dried over magnesium sulfate and the solvent evaporated under
vacuum. Purification by chromatography on silica, eluting with
40-100% EtOAc/hexane, afforded the title compound.
[1326] LC-MS Rt 1.01 mins [M+H].sup.+ 372.3 (Method
2minLowpHv03)
Step 5: Sodium
4-(5-(3-fluorophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d-
]pyrimidin-6(2H)-yl)-3-(hydroxymethyl)butanoate
[1327] Sodium hydroxide (2M, 2.00 mL, 4.0 mmol) was added to a
suspension of
5-(3-fluorophenyl)-1,3-dimethyl-6-((5-oxotetrahydrofuran-3-yl)methyl)--
1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (1.35 g, 3.6 mmol) in
ethanol (27 mL). The mixture was stirred at room temperature for 2
hours. The solvents were evaporated under vacuum to afford the
title compound.
[1328] LC-MS Rt 0.93 mins [M-Na+H].sup.+ 390.4 (Method
2minLowpHv03)
Step 6:
4-((tert-Butyldimethylsilyl)oxy)-3-((5-(3-fluorophenyl)-1,3-dimeth-
yl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)methyl)butano-
ic acid
[1329] A solution of tert-butyldimethylsilyl
trifluoromethanesulfonate (3.37 mL, 14.7 mmol) in DCM (5 mL) was
added dropwise to a suspension of sodium
4-(5-(3-fluorophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrol-
o[3,4-d]pyrimidin-6(2H)-yl)-3-(hydroxymethyl)butanoate (1.51 g, 3.7
mmol) and 2,6-lutidine (2.57 mL, 22.0 mmol) in DCM (20 mL) at
0.degree. C. The mixture was stirred at room temperature for 3
hours. Further portions of 2,6-lutidine (641 .mu.L, 5.5 mmol) and
tert-butyldimethylsilyl trifluoromethanesulfonate (843 .mu.L, 3.7
mmol) were added and the mixture stirred for a further 2 hours. The
reaction was diluted with DCM (50 mL), washed with water
(2.times.25 mL) and 1M HCl(aq) (1.times.25 mL), the organic phase
was dried over magnesium sulfate and evaporated under vacuum. The
residue was suspended in methanol (25 mL) and pyridine
hydrochloride (424 mg, 3.7 mmol) and water (2.5 mL) were added. The
mixture was stirred at room temperature for 2 hours then
concentrated under vacuum. The residue was dissolved in DCM (50 mL)
and washed with 1M HCl (aq) (2.times.25 mL) and brine (1.times.25
mL). The organic phase was dried over magnesium sulfate and
evaporated under vacuum. Trituration with diethyl ether afforded
the title compound.
[1330] LC-MS Rt 1.51 mins [M+H].sup.+ 504.5 (Method
2minLowpHv03)
Step 7:
4-((tert-Butyldimethylsilyl)oxy)-3-((5-(3-fluorophenyl)-1,3-dimeth-
yl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)methyl)-N-met-
hoxy-N-methylbutanamide
[1331] T3P.RTM. (2.62 mL, 4.5 mmol) was added to a suspension of
4-((tert-butyldimethylsilyl)oxy)-3-((5-(3-fluorophenyl)-1,3-dimethyl-2,4--
dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)methyl)butanoic
acid (1.13 g, 2.2 mmol), DIPEA (2.16 mL, 12.3 mmol) and
N,O-dimethylhydroxylamine hydrochloride (263 mg, 2.7 mmol) in DMF
(8 mL) at 0.degree. C. The mixture was stirred at room temperature
for 30 minutes. The reaction was diluted with ethyl acetate (40 mL)
and was washed with water (1.times.20 mL), 1M HCl(aq) (1.times.20
mL) and saturated NaHCO.sub.3(aq) (1.times.20 mL). The organic
phase was dried over magnesium sulfate and the solvent was removed
under vacuum. Purification by chromatography on silica, eluting
with 0-3% MeOH/DCM, afforded the title compound.
[1332] LC-MS Rt 1.62 mins [M+H].sup.+ 547.4 (Method
2minLowpHv03)
Step 8:
6-(2-(((tert-Butyldimethylsilyl)oxy)methyl)-4-(4-chlorothiazol-2-y-
l)-4-oxobutyl)-5-(3-fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-
-2,4(3H,6H)-dione
[1333] Isopropylmagnesium chloride lithium chloride complex
solution (1.3M, 1.6 mL, 2.1 mmol) was added to a solution of
4-((tert-butyldimethylsilyl)oxy)-3-((5-(3-fluorophenyl)-1,3-dimethyl-2,4--
dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-6(2H)-yl)methyl)-N-methoxy-N--
methylbutanamide (823 mg, 1.5 mmol) and 2-bromo-4-chlorothiazole
(Intermediate Q, step 2) (418 mg, 2.1 mmol) in THF (8 mL) at
0.degree. C. The mixture was stirred at room temperature for 1
hour. A further portion of isopropylmagnesium chloride lithium
chloride complex solution (1.3M, 116 .mu.L, 0.15 mmol) was added
the mixture stirred for a further 1 hour. The reaction was quenched
with saturated NH.sub.4Cl(aq) (5 mL), diluted with ethyl acetate
(40 mL) and washed with saturated NH.sub.4Cl(aq) (3.times.20 mL).
The organic phase was dried over magnesium sulfate and evaporated
under vacuum. Purification by chromatography on silica, eluting
with 20-40% EtOAc/hexane, afforded the title compound.
[1334] LC-MS Rt 1.80 mins [M+H].sup.+ 605.4 (Method
2minLowpHv03)
Step 9:
10-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-
-dimethyl-7,8-dihydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1335] Pyridine hydrochloride (516 mg, 4.5 mmol) was added to a
solution of
6-(2-(((tert-butyldimethylsilyl)oxy)methyl)-4-(4-chlorothiazol-2-yl)-4-
-oxobutyl)-5-(3-fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4-
(3H,6H)-dione (540 mg, 0.89 mmol) in methanol (14 mL) and water
(1.4 mL). The mixture was stirred at room temperature for 18 hours.
The reaction was concentrated under vacuum. The residue was diluted
with ethyl acetate (30 mL) and washed with 1M HCl (3.times.15 mL).
The organic phase was dried over magnesium sulfate, warmed at
35.degree. C. for 15 minutes, then evaporated under vacuum.
Trituration with diethyl ether afforded the title compound.
[1336] LC-MS Rt 1.17 mins [M+H].sup.+ 473.4 (Method
2minLowpHv03)
Step 10:
10-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,-
3-dimethyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1337] A mixture of
10-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8-dihydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione (310 mg,
0.66 mmol) and 10% platinum on carbon (310 mg) in ethanol (22 mL)
and tetrahydrofuran (22 mL) was stirred under an atmosphere of
hydrogen for 3 days. The reaction mixture was filtered and the
filtrates concentrated under vacuum. The residue was dissolved in
ethanol (22 mL) and tetrahydrofuran (22 mL), fresh 10% platinum on
carbon (310 mg) was added and the mixture further stirred under an
atmosphere of hydrogen for 5 days. The reaction mixture was
filtered and the filtrates evaporated under vacuum. The residue was
dissolved in 10 mL of 5% MeOH/DCM and the solution was treated with
silica-TMT (trimercaptotriazine functionalized silica, .about.1 g)
for 5 minutes, then filtered. The residue was washed with 5%
MeOH/DCM and the filtrates evaporated under vacuum. Purification by
chromatography on silica, eluting with 50-100% EtOAc/hexane
afforded the title compound as a mixture of diastereomers.
[1338] LC-MS Rt 1.15 mins [M+H].sup.+ 475.3 (Method
2minLowpHv03)
Step 10:
(8R,10R)-10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxym-
ethyl)-1,3-dimethyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H-
)-dione or
(8R,10S)-10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydrox-
ymethyl)-1,3-dimethyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,-
3H)-dione or
(8S,10R)-10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1-
,3-dimethyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
or
(8S,10S)-10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl-
)-1,3-dimethyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dio-
ne
[1339] The mixture of diastereomers was separated by SFC
chromatographic resolution using the following combination of two
SFC conditions in series to afford the title compounds as single
diastereomers.
[1340] The first separation resolves Diastereoisomer 2 and another
diastereoisomer.
[1341] The second separation resolves Diastereoisomer 1 and
Diasteroisomer 3
[1342] Separation condition 1:
[1343] Column: Chiralpak OJ-H 250.times.10 mm, 5 .mu.m
[1344] Mobile Phase: 40% MeOH/60% CO2
[1345] Flow: 10 mL/min
[1346] Detection: UV @ 220 nm
Example 23b
Diastereomer 2 of
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1347] Rt=5.70mins (First separation) 1H NMR (400 MHz, DMSO-d6)
.delta. 7.65 (1H, s), 7.53 (1H, mult), 7.41-7.28 (3H, mult), 5.29
(1H, t), 4.75 (1H, t), 3.89 (1H, mult), 3.74 (1H, t), 3.42 (1H,
mult), 3.18-3.14 (6H, mult), 2.03 (1H, mult), 1.72 (1H, mult).
[1348] LC-MS Rt 1.15 mins [M+H].sup.+ 475.4 (Method
2minLowpHv03)
Another Diastereomer of
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1349] Rt=2.42mins was isolated (First separation)
Second Separation Resolves Diastereomer 1 and Diastereomer 3
[1350] Column: Chiralcel OD-H 250.times.10 mm, 5 .mu.m
[1351] Mobile Phase: 30% MeOH/70% CO2
[1352] Flow: 10 mL/min
[1353] Detection: UV @ 220 nm
Example 23a
Diastereomer 1 of
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1354] Rt=6.02mins (Second separation)
[1355] 1H NMR (400 MHz, DMSO-d6) .delta. 7.63 (1H, s), 7.53 (1H,
mult), 7.36-7.29 (3H, mult), 5.36 (1H, mult), 4.69 (1H, br t), 3.98
(1H, dd), 3.62 (1H, t), 3.29 (3H, s), 3.16 (3H, s), 2.35 (1H, d),
2.02 (1H, mult), 1.91 (1H, mult).
[1356] LC-MS Rt 1.15 mins [M+H].sup.+ 475.3 (Method
2minLowpHv03)
Example 23c
Diastereomer 3 of
10-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-8-(hydroxymethyl)-1,3-dimeth-
yl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1357] Rt=7.03 mins (Second separation)
[1358] 1H NMR (400 MHz, DMSO-d6) .delta. 7.63 (1H, s), 7.53 (1H,
mult), 7.36-7.28 (3H, mult), 5.36 (1H, mult), 4.69 (1H, br t), 3.98
(1H, dd), 3.62 (1H, t), 3.30 (3H, s), 3.16 (3H, s), 2.35 (1H, d),
2.02 (1H, mult), 1.91 (1H, mult).
[1359] LC-MS Rt 1.15 mins [M+H].sup.+ 475.5 (Method
2minLowpHv03)
[1360] The following listed examples were prepared in a similar
manner to Example 23a, 23b and 23c replacing
5-(3-fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-di-
one in step 4 with the appropriate material (preparation described
hereinafter). The mixtures of diastereomers were separated by SFC
chromatographic resolution under the listed conditions to afford
the title compounds as single diastereomers.
Example 23.1a, 23b and 23c
3-((8R,10R)-10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-d-
ioxo-1,2,3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
or
3-((8S,10R)-10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-d-
ioxo-1,2,3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
or
3-((8R,10S)-10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-d-
ioxo-1,2,3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
or
3-((8S,10S)-10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-d-
ioxo-1,2,3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
##STR00139## ##STR00140##
[1362] Separation conditions:
[1363] Column: LUX 250.times.10 mm, 5 .mu.m
[1364] Mobile Phase: 50% MeOH/50% CO2
[1365] Flow: 10 mL/min
[1366] Detection: UV @ 220 nm
Example 23.1a
Diastereomer 1 of
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-1,2-
,3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
[1367] Rt=17.26 mins
[1368] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.79-7.72 (3H, mult),
7.63 (1H, t), 7.04 (1H, s), 5.18 (1H, mult), 4.14 (1H, dd), 3.73
(1H, dd), 3.65-3.55 (2H, mult), 3.45 (3H, s), 3.36 (3H, s), 2.63
(1H, d), 2.21-2.10 (2H, mult).
[1369] LC-MS Rt 1.11 mins [M+H].sup.+ 482.5 (Method
2minLowpHv03)
Example 23.1 b
Diastereomer 2 of
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-1,2-
,3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
[1370] Rt=9.43 mins
[1371] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.80-7.74 (3H, mult),
7.62 (1H, t), 7.07 (1H, s), 5.10 (1H, t), 4.02 (1H, dd), 3.75-3.66
(2H, mult), 3.61 (1H, dd), 3.37-3.35 (6H, mult), 2.66 (1H, mult),
2.26 (1H, mult), 2.17-2.08 (1H, mult).
[1372] LC-MS Rt 1.10 mins [M+H].sup.+ 482.4 (Method
2minLowpHv03)
Example 23.1c
Diastereomer 3 of
3-(10-(4-chlorothiazol-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-1,2-
,3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-5-yl)benzonitrile
[1373] Rt=14.67 mins
[1374] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.80-7.74 (3H, mult),
7.62 (1H, t), 7.07 (1H, s), 5.10 (1H, t), 4.02 (1H, dd), 3.75-3.67
(2H, mult), 3.61 (1H, dd), 3.37-3.34 (6H, mult), 2.66 (1H, mult),
2.26 (1H, mult), 2.16-2.08 (1H, mult).
[1375] LC-MS Rt 1.10 mins [M+H].sup.+ 482.4 (Method
2minLowpHv03)
[1376] Diastereomer 4 was isolated at Rt=27.87 mins
Example 24
(7S)-7-((2H-1,2,3-triazol-2-yl)methyl)-1,3-dimethyl-10-(5-methylfuran-2-yl-
)-5-phenyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
or
(7R)-7-((2H-1,2,3-triazol-2-yl)methyl)-1,3-dimethyl-10-(5-methylfuran-2-yl-
)-5-phenyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
##STR00141##
[1377] Step 1: (S)-Dibenzyl
2-(1,3-dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-
-6(2H)-yl)pentanedioate
[1378]
5-benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
(Intermediate C) (2.04 g, 6.05 mmol), (S)-dibenzyl
2-aminopentanedioate (commercially available) (4.53 g, 9.08 mmol)
and triethylamine (3.37 mL, 24.20 mmol) were suspended in dioxane
(30 mL) and the mixture was heated at reflux for 2 hours. The
mixture was cooled to room temperature and evaporated under vacuum.
The residue was partitioned between DCM and 0.1M HCl(aq) and the
phases separated. The organic phase was passed through a
hydrophobic frit. Purification by chromatography on silica, eluting
with 20-60% EtOAc/hexane, afforded the title compound.
[1379] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.50-7.25 (15H, mult),
6.44 (1H, s), 5.24 (1H, d), 5.20 (1H, d), 4.99 (1H, d), 4.95 (1H,
mult), 4.92 (1H, mult), 3.39 (3H, s), 3.35 (3H, s), 2.48 (1H,
mult), 2.31 (1H, mult), 2.20 (1H, mult), 2.09 (1H, mult).
[1380] LC-MS Rt 1.33 mins [M+H].sup.+ 566.4 (Method
2minLowpHv01)
Step 2:
(S)-2-(1,3-Dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4--
d]pyrimidin-6(2H)-yl)pentanedioic acid
[1381] A suspension of (S)-dibenzyl
2-(1,3-dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-
-6(2H)-yl)pentanedioate (3 g, 5.30 mmol) and 10% palladium on
carbon (0.564 g, 0.530 mmol) in ethanol (100 mL) was stirred under
a hydrogen atmosphere for 140 mins. The mixture was filtered
through a Celite.RTM. cartridge (10 g, filter material) and the
residue rinsed well with methanol and 7N NH.sub.3/methanol. The
combined filtrates were evaporated under vacuum to afford the title
compound.
[1382] 1H NMR (MHz, DMSO-d6) .delta. 7.45-7.38 (5H, mult), 6.90
(1H, s), 4.33 (1H, t), 3.33 (3H, s), 3.15 (3H, s), 2.31 (1H, mult),
2.11-1.87 (3H, mult).
[1383] LC-MS Rt 0.78 mins [M+H].sup.+ 386.3 (Method
2minLowpHv01)
Step 3:
(S)-1,3-Dimethyl-2,4,10-trioxo-5-phenyl-1,2,3,4,7,8,9,10-octahydro-
pyrimido[4,5-a]indolizine-7-carboxylic acid
[1384]
(S)-2-(1,3-Dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d-
]pyrimidin-6(2H)-yl)pentanedioic acid (2.44 g, 6.33 mmol) and
polyphosphoric acid (4.85 ml, 6.33 mmol) were combined and heated
at 110.degree. C. for 85 mins. The reaction mixture was cooled to
.about.50.degree. C. and diluted with water to give a fluid
mixture. The mixture was then cooled to room temperature, further
diluted with water and extracted with chloroform (5.times.). The
combined organic extracts were passed through a hydrophobic frit
and evaporated under vacuum to afford the title compound.
[1385] LC-MS Rt 0.88 mins [M-H]-366.2 (Method 2minLowpHv01)
Step 4: (S)-Methyl
1,3-dimethyl-2,4,10-trioxo-5-phenyl-1,2,3,4,7,8,9,10-octahydropyrimido[4,-
5-a]indolizine-7-carboxylate
[1386]
(S)-1,3-Dimethyl-2,4,10-trioxo-5-phenyl-1,2,3,4,7,8,9,10-octahydrop-
yrimido[4,5-a]indolizine-7-carboxylic acid (1.85 g, 5.04 mmol),
potassium carbonate (2.088 g, 15.11 mmol) and dimethyl sulfate
(1.444 mL, 15.11 mmol) were combined in acetone (50 mL) and water
(5 mL). The mixture was heated at reflux for 1.5 hours, then cooled
to room temperature and evaporated under vacuum. The residue was
partitioned between DCM and water and the phases separated. The
organic phase was passed through a hydrophobic frit and evaporated
under vacuum. Purification by chromatography on silica, eluting
with 40-80% EtOAc/hexane, afforded the title compound.
[1387] LC-MS Rt 1.01 mins [M+H].sup.+ 382.3 (Method
2minLowpHv01)
Step 5: (7S)-Methyl
10-hydroxy-1,3-dimethyl-2,4-dioxo-5-phenyl-1,2,3,4,7,8,9,10-octahydropyri-
mido[4,5-a]indolizine-7-carboxylate
[1388] Sodium borohydride (0.179 g, 4.72 mmol) was added slowly to
a suspension of
(S)-1,3-dimethyl-2,4,10-trioxo-5-phenyl-1,2,3,4,7,8,9,10-octahydropyrimid-
o[4,5-a]indolizine-7-carboxylic acid (1.8 g, 4.72 mmol) in methanol
(40 mL). The mixture was stirred at room temperature for 30 mins.
The reaction was quenched with saturated NaHCO.sub.3(aq) and
extracted with DCM. The organic phase was passed through a
hydrophobic frit and evaporated under vacuum to afford the title
compound.
[1389] LC-MS Rt 0.94 mins [M+H].sup.+ 384.3 (Method
2minLowpHv01)
Step 6: (7S)-Methyl
1,3-dimethyl-10-(5-methylfuran-2-yl)-2,4-dioxo-5-phenyl-1,2,3,4,7,8,9,10--
octahydropyrimido[4,5-a]indolizine-7-carboxylate
[1390] Gold (III) chloride (0.142 g, 0.469 mmol) was added to a
solution of (7S)-methyl
10-hydroxy-1,3-dimethyl-2,4-dioxo-5-phenyl-1,2,3,4,7,8,9,10-octahydropyri-
mido[4,5-a]indolizine-7-carboxylate (1.8 g, 4.69 mmol) and
2-methylfuran (0.466 mL, 5.16 mmol) in acetonitrile (30 mL). The
mixture was stirred at room temperature for 25 mins, then quenched
with saturated NaHCO.sub.3(aq). The mixture was extracted with DCM,
the organic phase passed through a hydrophobic frit and evaporated
under vacuum. Purification by chromatography on silica, eluting
with 20-40% EtOAc/hexane, afforded the title compound.
[1391] LC-MS Rt 1.22 mins [M+H].sup.+ 448.5 (Method
2minLowpHv01)
Step 7:
(7S)-7-(Hydroxymethyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-5-phen-
yl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1392] Lithium aluminium hydride (1M in THF, 3.44 mL, 3.44 mmol)
was added to a solution of (7S)-methyl
1,3-dimethyl-10-(5-methylfuran-2-yl)-2,4-dioxo-5-phenyl-1,2,3,4,7,8,9,10--
octahydro pyrimido[4,5-a]indolizine-7-carboxylate (1.4 g, 3.13
mmol) in THF (30 mL) at 0.degree. C. The mixture was stirred at
room temp. for 1 hour. The reaction mixture was quenched at
0.degree. C. by the slow dropwise addition of water (1 mL), stirred
vigourously at 0.degree. C. for 5 mins, then diluted with EtOAc and
further stirred vigourously for 20 mins. The mixture was filtered,
rinsing the residue with EtOAc. The filtrates were passed through a
hydrophobic frit and evaporated under vacuum to afford the title
compound.
[1393] LC-MS Rt 1.10 mins [M+H].sup.+ 420.3 (Method
2minLowpHv01)
Step 8:
((7S)-1,3-Dimethyl-10-(5-methylfuran-2-yl)-2,4-dioxo-5-phenyl-1,2,-
3,4,7,8,9,10-octahydropyrimido[4,5-a]indolizin-7-yl)methyl
methanesulfonate
[1394] Methanesulfonyl chloride (0.483 mL, 6.20 mmol) was added to
a solution of
(7S)-7-(hydroxylmethyl)-1,3-dimethyl-10-(5-methylfuran-2-yl)-5-phenyl-7,8-
,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione (1.3 g,
3.10 mmol) and triethylamine (1.296 mL, 9.30 mmol) in DCM (30 mL).
The mixture was stirred at room temp. for 105 mins. The reaction
was quenched with saturated NaHCO.sub.3(aq) and extracted with DCM.
The organic extract was passed through a hydrophobic frit and
evaporated under vacuum. Purification by chromatography on silica,
eluting with 40-75% EtOAc/hexane, afforded the title compound.
[1395] LC-MS Rt 1.20 mins [M+H].sup.+ 498.3 (Method
2minLowpHv01)
Step 9:
(7S)-7-((2H-1,2,3-triazol-2-yl)methyl)-1,3-dimethyl-10-(5-methylfu-
ran-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-
-dione
[1396]
((7S)-1,3-Dimethyl-10-(5-methylfuran-2-yl)-2,4-dioxo-5-phenyl-1,2,3-
,4,7,8,9,10-octahydro pyrimido[4,5-a]indolizin-7-yl)methyl
methanesulfonate (200 mg, 0.402 mmol), potassium carbonate (167 mg,
1.206 mmol) and 1,2,3-triazole (0.070 mL, 1.206 mmol) were combined
in DMF (3 mL) and the mixture heated at 120.degree. C. under
microwave irradiation for 1 hour. The mixture was partitioned
between EtOAc and water, and extracted with EtOAc (2.times.). The
organic phase was washed with brine (4.times.), dried over sodium
sulfate and evaporated under vacuum. Purification by chromatography
on silica, eluting with 20-100% EtOAc/hexane, afforded the title
compound as a mixture of diastereomers
[1397] LC-MS Rt 1.34 mins [M+H].sup.+ 471.3 (Method
2minLowpHv04)
Step 10:
(7S)-7-((2H-1,2,3-triazol-2-yl)methyl)-1,3-dimethyl-10-(5-methylf-
uran-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H-
)-dione or
(7R)-7-((2H-1,2,3-triazol-2-yl)methyl)-1,3-dimethyl-10-(5-methy-
lfuran-2-yl)-5-phenyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,-
3H)-dione
[1398] The diastereomeric mixture of
(7S)-7-((2H-1,2,3-triazol-2-yl)methyl)-1,3-dimethyl-10-(5-methylfuran-2-y-
l)-5-phenyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
was separated by SFC under the following conditions to afford the
title compound as a single disatereomers.
[1399] Separation conditions:
[1400] Column: Chiralcel OD-H 250.times.10 mm.times.5 um @
34.5.degree. C., UV @ 220 nm
[1401] Eluent: 25% MeOH/75% CO2
[1402] Flow: 10 mL/min
[1403] Detection: UV @ 220 nm
Example 24
Enantiomer 1 of
(7S)-7-((2H-1,2,3-triazol-2-yl)methyl)-1,3-dimethyl-10-(5-methylfuran-2-y-
l)-5-phenyl-7,8,9,10-tetrahydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1404] Rt=12.29 mins
[1405] LC-MS Rt 1.37 mins [M+H].sup.+ 471.6 (Method
2minLowpHv03)
[1406] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.52-7.39 (5H, mult),
7.20 (2H, s), 5.78 (1H, mult), 5.62 (1H, mult), 4.99 (1H, mult),
4.68 (1H, mult), 4.24 (1H, mult), 4.22 (1H, mult), 3.40 (3H, s),
3.27 (3H, s), 2.25 (1H, mult), 2.21 (3H, s), 2.07 (1H, mult),
1.81-1.62 (2H, mult).
[1407] The second diastereomer was isolated at Rt=5.93 mins.
Example 25
2-(10-(5-Chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-hexahydro-1-
H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)thiazole-4-carboxylic
acid
##STR00142##
[1409] Ethyl
2-(10-(5-chlorofuran-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,10-hexahydro--
1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-5-yl)thiazole-4-carboxyla-
te (Example 15 Step 1)(72 mg, 0.142 mmol) and lithium hydroxide
monohydrate (5.96 mg, 0.142 mmol) were suspended in THF (5 mL) and
the mixture stirred at room temperature for 24 hours. Two portions
of lithium hydroxide monohydrate (5.96 mg, 0.142 mmol) were added
during this period. Water (2 mL) was then added and the mixture
stirred for a further 30 mins. The resulting mixture was evaporated
under vacuum. The residue was partitioned between water (20 ml) and
DCM (20 mL) and brought to pH 1 with HCl(aq). The phases were
separated and the aqueous phase was extracted with DCM (2.times.20
ml). The combined organic extracts were passed through a
hydrophobic frit and evaporated under vacuum. The residue was
re-suspended in DCM (5 ml) and loaded to a Isolute PE-AX ion
exchange cartridge (1 g). The cartridge was eluted with DCM (50
ml), 1:1 MeOH:DCM (20 ml) and 4 M HCl/dioxane. The acidic eluent
was evaporated under vacuum and the residue triturated with
TBME:iso-hexane to afford the title compound.
[1410] 1H NMR (400 MHz, DMSO-d6) .delta. 13.15 (1H, br s), 8.66
(1H, s), 6.42 (1H, d), 6.22 (1H, s), 6.20 (1H, dd), 4.61 (1H, dt),
4.45 (1H, m), 3.45 (3H, s), 3.21 (3H, s), 3.10-2.94 (2H, m)
[1411] LC-MS Rt 1.25 mins [M+H].sup.+ 479.2 (Method
2minLowpHv03)
Example 26a, 26b and 26c
N-(((8R,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)methanesulfonamide or
N-(((8R,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)methanesulfonamide or
N-(((8S,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)methanesulfonamide or
N-(((8S,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8)methyl)methanesulfonamide
##STR00143## ##STR00144##
[1413] Methanesulfonyl chloride (0.112 mL, 1.444 mmol) was added to
a solution of
8-(aminomethyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-
-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazine-2,4(3H,-
10H)-dione (Example 19) (69 mg, 0.144 mmol) and DIPEA (0.504 mL,
2.89 mmol) in DCM (2 mL). The mixture was stirred at room
temperature for 16 hours, then diluted with ethyl acetate (20 ml)
and washed with water (20 ml), 10% w/w citric acid (aq) (20 ml) and
brine (20 ml). The organic layer was passed through a hydrophobic
frit and evaporated under vacuum. Purification by mass-directed
HPLC under the following conditions afforded two racemic
diastereomers.
[1414] Column: XSelect CSH Prep C18 column, 30.times.100 mm, 5
um.
[1415] Mobile phase: A=0.1% DEA in water, B=0.1% DEA in MeCN
[1416] Gradient:
[1417] 0.0-0.5 min: 30% B 30 mL/min
[1418] 0.5-1.0 min: 30% B 30-50 mL/min
[1419] 1.0-7.2 min: 30-70% B, 7.2-7.3 min: 70-98% B, 7.3-9.4 min:
98% B
[1420] 9.4-9.5 30% B 50 mL/min
Diastereomer pair 1 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)methanesulfonamide
[1421] LC-MS Rt 1.21 min [M+H].sup.+ 556.5 (Method
2minLowpHv03)
Diastereomer pair 2 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)methanesulfonamide
[1422] LCMS Rt 1.31 min [M+H].sup.+ 556.5 (Method 2minLowpHv03)
[1423] Diastereomer pair 1 was resolved into enantiomers by SFC
chromatographic resolution under the following conditions to afford
the listed compound.
[1424] Column: OJ CHIRALCEL, 250.times.10 mm, 5 um @ 35.degree.
C.
[1425] Mobile phase: 50% Methanol/50% CO2
[1426] Flow: 10 ml/min
[1427] Detection: UV @ 220 nm
Example 26b
Diastereomer 2 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)methanesulfonamide
[1428] Rt=3.32 mins
[1429] LC-MS Rt 1.23 min [M+H].sup.+ 556.3 (Method
2minLowpHv03)
[1430] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.16 (s, 1H), 6.14 (d,
1H), 6.08 (d, 1H), 5.75 (s, 1H), 5.49 (dd, 1H), 5.26 (s, 1H), 4.21
(dd, 1H), 3.73 (m, 1H), 3.59 (s, 3H), 3.43 (s, 5H), 3.00 (s, 3H),
2.57 (s, 3H).
[1431] Diastereomer pair 2 was resolved into enantiomers by SFC
chromatographic resolution under the following conditions to afford
the listed compounds;
[1432] Column: Chiralpak AD-H 250.times.10 mm, 5 um @ 35 deg C.
[1433] Mobile phase: 50% Methanol/50% CO2
[1434] Flow: 10 ml/min
[1435] Detection: UV @ 220 nm
Example 26a
Diastereomer 1 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)methanesulfonamide
[1436] Rt=5.69 mins
[1437] 1H NMR (400 MHz, CDCl.sub.3) .delta. 8.23 (s, 1H), 7.22 (s,
1H), 6.26 (dd, 1H), 6.13 (d, 1H), 5.79 (s, 1H), 5.41 (dd, 1H), 4.06
(dd, 1H), 3.85 (dd, 1H), 3.72 (s, 3H), 3.43 (s, 3H), 3.29 (m, 1H),
2.92 (s, 3H), 2.62 (s, 4H), 2.59 (m, 1H).
[1438] LC-MS Rt 1.32 min [M+H].sup.+ 556.3 (Method
2minLowpHv03)
Example 26c
Diastereomer 3 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)methanesulfonamide
[1439] Rt=4.45 mins
[1440] 1H NMR (400 MHz, CDCl.sub.3) .delta. 8.22 (s, 1H), 7.22 (s,
1H), 6.26 (dd, 1H), 6.13 (d, 1H), 5.79 (s, 1H), 5.41 (dd, 1H), 4.06
(d, 1H), 3.85 (dd, 1H), 3.72 (s, 3H), 3.43 (s, 3H), 3.29 (d, 1H),
2.92 (s, 3H), 2.62 (s, 3H), 2.59 (m, 1H).
[1441] LC-MS Rt 1.30 min [M+H].sup.+ 556.3 (Method
2minLowpHv03)
[1442] The following listed examples were prepared in a similar
manner to Example 26a,b and c by replacing methanesulfonyl chloride
with the appropriate acyl chloride. Purification and
chromatographic conditions as described.
Example 26.1a, 26.1b and 26.1c
N-(((8R,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)-2-methoxyacetamide or
N-(((8R,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)-2-methoxyacetamide or
N-(((8S,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)-2-methoxyacetamide or
N-(((8S,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)-2-methoxyacetamide
##STR00145## ##STR00146##
[1444] Separation of the racemic diastereomers by achiral SFC:
[1445] Column: Princeton DEAP 21.2.times.150 mm, 5 um
[1446] Mobile Phase: A=Methanol, B.dbd.CO2
[1447] Flow: 100 mL/min
[1448] Gradient:
[1449] 0-0.5 min: 1% A
[1450] 0.5-8.0 min: 1-10% A
[1451] 8.0-8.1 min: 10-40% A
[1452] 8.1-8.6 min: 40% A
[1453] 8.6-8.7 min: 40-1% A
Diastereomer pair 1 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)-2-methoxyacetamide
[1454] LC-MS Rt=1.21 mins [M+H].sup.+ 550.5 (Method
2minLowpHv03)
[1455] Diastereomer pair 2 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)-2-methoxyacetamide
[1456] LC-MS Rt=1.21 mins [M+H].sup.+ 550.5 (Method
2minLowpHv03)
[1457] Diastereomer pair 1 was separated by SFC chromatographic
resolution under the following conditions to afford the listed
compound;
[1458] Column: Chiralcel OJ-H 250.times.10 mm, 5 um @ 35 deg C.
[1459] Mobile phase: 45% Methanol/55% CO2
[1460] Flow: 10 ml/min
[1461] Detection: UV @ 220 nm
Example 26.1b
Diastereomer 2 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)-2-methoxyacetamide
[1462] Rt=2.88 mins
[1463] 1H NMR (400 MHz, Chloroform-d) .delta. 7.12 (s, 1H), 6.92
(t, 1H), 6.22 (dd, 1H), 6.13 (d, 1H), 5.77 (d, 1H), 5.70 (dd, 1H),
3.89 (s, 2H), 3.87-3.82 (m, 1H), 3.72 (m, 1H), 3.66 (s, 3H), 3.66
(m, 1H), 3.55 (m, 1H), 3.43 (s, 3H), 3.43 (s, 3H), 2.52 (s, 3H)
[1464] LC-MS Rt 1.31 min [M+H].sup.+ 550.4 (Method
2minLowpHv03)
[1465] Diastereomer pair 2 was separated by SFC chromatographic
resolution under the following conditions to afford the listed
compounds;
[1466] Column: Chiralpak AD-H 250.times.10 mm, 5 um @ 35 deg C.
[1467] Mobile phase: 40% Isopropanol/60% CO2
[1468] Flow: 10 ml/min
[1469] Detection: UV @ 220 nm
Example 26.1a
Diastereomer 1 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)-2-methoxyacetamide
[1470] Rt=5.21 mins
[1471] 1H NMR (400 MHz, CDCl3) .delta. 7.18 (m, 1H), 7.16 (s, 1H),
6.24 (dd, 1H), 6.12 (d, 1H), 5.77 (d, 1H), 5.49 (dd, 1H), 4.19 (dd,
1H), 3.93 (d, 1H), 3.84 (d, 1H), 3.76 (m, 1H), 3.70 (s, 3H), 3.44
(s, 3H), 3.39 (m, 1H), 3.36 (s, 3H), 3.03 (m, 1H), 2.57 (s, 3H)
[1472] LC-MS Rt 1.23 min [M+H]+ 550.2 (Method 2minLowpHv03)
Example 26.1c
Diastereomer 3 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)-2-methoxyacetamide
[1473] Rt=3.55 mins
[1474] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.18 (m, 1H), 7.16 (s,
1H), 6.24 (dd, 1H), 6.12 (d, 1H), 5.77 (d, 1H), 5.49 (dd, 1H), 4.19
(dd, 1H), 3.93 (d, 1H), 3.84 (d, 1H), 3.76 (m, 1H), 3.70 (s, 3H),
3.44 (s, 3H), 3.39 (m, 1H), 3.36 (s, 3H), 3.03 (m, 1H), 2.57 (s,
3H)
[1475] LC-MS Rt 1.23 min [M+H]+ 550.2 (Method 2minLowpHv03)
Example 26.2a and 26.2b
Methyl
(((8S,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol--
2-yl)-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c-
][1,4]thiazin-8-yl)methyl) carbamate or
Methyl
(((8S,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol--
2-yl)-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c-
][1,4]thiazin-8-yl)methyl) carbamate or
Methyl
(((8R,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol--
2-yl)-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c-
][1,4]thiazin-8-yl)methyl) carbamate or
Methyl
(((8R,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol--
2-yl)-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c-
][1,4]thiazin-8-yl)methyl) carbamate
##STR00147## ##STR00148##
[1477] The mixture of diastereomers was separated into two racemic
diastereomers by mass-directed HPLC under the following
conditions;
[1478] Column: XSelect CSH Prep C18 column, 30.times.100 mm, 5
um.
[1479] Mobile phase: A=0.1% DEA in water, B=0.1% DEA in MeCN
[1480] Gradient:
[1481] 0.0-0.5 min: 40% B 30 mL/min
[1482] 0.5-1.0 min: 40% B 30-50 mL/min
[1483] 1.0-7.2 min: 40-80% B, 7.2-7.3 min: 80-98% B, 7.3-9.4 min:
98% B 9.4-9.5 40% B 50 mL/min
Diastereomer pair 1 of methyl
((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dioxo-
-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-8-
-yl)methyl)carbamate
[1484] LC-MS Rt 1.28 mins [M+H].sup.+ 536.2 (Method
2minLowpHv03)
Diastereomer pair 2 of methyl
((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dioxo-
-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-8-
-yl)methyl)carbamate
[1485] LC-MS Rt 1.36 mins [M+H].sup.+ 536.2 (Method
2minLowpHv03)
[1486] Diastereomer pair 1 was resolved into single diastereomers
by SFC chromatographic resolution under the following conditions to
give the listed compound;
[1487] Column: OJ CHIRALCEL, 250.times.10 mm, 5 um @ 35.degree.
C.
[1488] Mobile phase: 50% Methanol/50% CO2
[1489] Flow: 10 ml/min
[1490] Detection: UV @ 220 nm
Example 26.2a
Diastereomer 1 of methyl
((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dioxo-
-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-8-
-yl)methyl)carbamate
[1491] Rt=3.46 mins
[1492] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.12 (d, 1H), 6.13 (d,
2H), 5.74 (s, 1H), 5.66 (dd, 1H), 5.22 (s, 1H), 3.90 (dd, 1H), 3.70
(s, 1H), 3.68 (s, 3H), 3.64 (s, 3H), 3.54 (m, 1H), 3.44 (m, 1H),
3.43 (s, 3H), 2.53 (s, 3H)
[1493] LC-MS Rt 1.29 min [M+H].sup.+ 536.3 (Method
2minLowpHv03)
[1494] Diastereomer pair 2 was resolved into single diastereomers
by SFC chromatographic resolution under the following conditions to
give the listed compound;
[1495] Column: Chiralpak AD-H, 250.times.10 mm, 5 um @ 35 deg
C.
[1496] Mobile phase: 40% Methanol/60% CO2
[1497] Flow: 10 ml/min
[1498] Detection: UV @ 220 nm
Example 26.2b
Diastereomer 2 of methyl
((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dioxo-
-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-8-
-yl)methyl)carbamate
[1499] Rt=6.10 mins
[1500] 1H NMR (400 MHz, CDCl3) .delta. 7.65 (s, 1H), 7.20 (s, 1H),
6.25 (d, 1H), 6.12 (d, 1H), 5.77 (s, 1H), 5.41 (d, 1H), 3.99 (d,
1H), 3.71 (s, 6H), 3.69 (m, 1H), 3.59 (d, 1H), 3.43 (s, 3H), 2.60
(s, 3H), 2.55 (dd, 1H)
[1501] LC-MS Rt 1.36 min [M+H].sup.+ 536.2 (Method
2minLowpHv03)
Example 26.3a, 26.3b, 26.3c and 26d
N-(((8S,10R)-10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)acetamide or
N-(((8S,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)acetamide or
N-(((8R,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)acetamide or
N-(((8R,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)acetamide
##STR00149## ##STR00150##
[1503] The mixture of diastereomers was separated by single-step
SFC chromatographic resolution under the listed conditions to give
the title compounds.
[1504] Column: Chiralpak AD-H 250.times.10 mm, 5 um @ 35 deg C.
[1505] Mobile phase: 30% Isopropanol/70% CO2
[1506] Flow: 10 ml/min
[1507] Detection: UV @ 220 nm
Example 26.3a
Diastereomer 1 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)acetamide
[1508] Rt=9.38 mins
[1509] 1H NMR (400 MHz, CDCl3) .delta. 7.14 (s, 1H), 6.14 (d, 1H),
6.12 (dd, 1H), 5.99 (t, 1H), 5.75 (d, 1H), 5.57 (dd, 1H), 3.85 (dd,
1H), 3.74 (m, 1H), 3.64 (m, 1H), 3.63 (s, 3H), 3.45 (dd, 1H), 3.42
(s, 3H), 2.53 (s, 3H), 1.97 (s, 3H)
[1510] LC-MS Rt 1.23 min [M+H].sup.+ 520.4 (Method
2minLowpHv03)
Example 26.3b
Diastereomer 2 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)acetamide
[1511] Rt=12.03 mins
[1512] 1H NMR (400 MHz, CDCl3) .delta. 7.59 (dd, 1H), 7.23 (s, 1H),
6.25 (dd, 1H), 6.11 (d, 1H), 5.78 (s, 1H), 5.37 (dd, 1H), 4.05 (m,
1H), 3.72 (m, 2H), 3.71 (s, 3H), 3.42 (s, 3H), 2.59 (s, 3H), 2.45
(m, 1H), 2.06 (s, 3H)
[1513] LC-MS Rt 1.19 min [M+H].sup.+ 520.3 (Method
2minLowpHv03)
Example 26.3c
Diastereomer 3 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)acetamide
[1514] Rt=4.82 mins
[1515] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.14 (s, 1H), 6.14 (d,
1H), 6.12 (dd, 1H), 6.01 (t, 1H), 5.75 (d, 1H), 5.57 (dd, 1H), 3.85
(dd, 1H), 3.74 (m, 1H), 3.64 (m, 1H), 3.63 (s, 3H), 3.45 (dd, 1H),
3.42 (s, 3H), 2.53 (s, 3H), 1.97 (s, 3H)
[1516] LC-MS Rt 1.15 min [M+H].sup.+ 520.3 (Method
2minLowpHv03)
Example 26.3d
Diastereomer 4 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)acetamide
[1517] Rt=5.82 mins
[1518] 1H NMR (400 MHz, CDCl3) .delta. 7.59 (dd, 1H), 7.23 (s, 1H),
6.25 (dd, 1H), 6.11 (d, 1H), 5.78 (s, 1H), 5.37 (dd, 1H), 4.05 (m,
1H), 3.72 (m, 2H), 3.71 (s, 3H), 3.42 (s, 3H), 2.59 (s, 3H), 2.45
(m, 1H), 2.06 (s, 3H)
[1519] LC-MS Rt 1.20 min [M+H].sup.+ 520.5 (Method
2minLowpHv03)
Example 26.4a and 26.4b
N-(((8S,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)benzamide or
N-(((8S,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)benzamide or
N-(((8R,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)benzamide or
N-(((8R,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-
-2,4-dioxo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazin-8-yl)methyl)benzamide
##STR00151## ##STR00152##
[1521] The mixture of diastereomers was separated by single-step
SFC chromatographic resolution under the listed conditions to give
the title compounds.
[1522] Column: Phenomenex LUX-A2 250.times.10 mm, 5 um @ 35 deg
C.
[1523] Mobile phase: 50% Isopropanol/50% CO2
[1524] Flow: 10 ml/min
[1525] Detection: UV @ 220 nm
Example 26.4a
Diastereomer 1 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)benzamide
[1526] Rt=6.94 mins
[1527] LC-MS Rt 1.38 min [M+H]+ 582.4 (Method 2minLowpHv03)
[1528] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.57 (d, 2H), 7.47 (t,
1H), 7.41 (d, 1H), 7.33 (t, 2H), 7.02 (s, 1H), 6.18 (dd, 1H), 6.03
(d, 1H), 5.73 (s, 1H), 5.48 (dd, 1H), 4.03 (dd, 1H), 3.83 (m, 1H),
3.70 (m, 1H), 3.64 (s, 3H), 3.35 (s, 3H), 2.70 (ddd, 1H), 1.99 (d,
3H)
Example 26.4b
Diastereomer 2 of
N-((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dio-
xo-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-
-8-yl)methyl)benzamide
[1529] Rt=6.94 mins
[1530] LC-MS Rt 1.36 min [M+H]+ 582.4 (Method 2minLowpHv03)
[1531] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.71 (d, 2H), 7.55 (t,
1H), 7.47 (dd, 2H), 7.10 (s, 1H), 6.63 (t, 1H), 6.11 (dd, 1H), 6.02
(d, 1H), 5.79 (d, 1H), 5.66 (d, 1H), 3.99 (dd, 1H), 3.86 (m, 1H),
3.74 (m, 2H), 3.63 (s, 3H), 3.42 (s, 3H), 2.29 (s, 3H)
Example 27a and 27b
(7R,9R)-9-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-7-(hydroxymethyl)-1,3--
dimethyl-8,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
and
(7S,9R)-9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-7-(hydroxymethyl)-1,3--
dimethyl-8,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00153##
[1532] Step 1:
7-((4-Chlorothiazol-2-yl)(hydroxy)methyl)-5-(3-fluorophenyl)-1,3-dimethyl-
-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1533] The title compound was prepared by an analogous method to
7-((4-chlorothiazol-2-yl)(hydroxy)methyl)-1,3-dimethyl-5-phenyl-1H-pyrrol-
o[3,4-d]pyrimidine-2,4(3H,6H)-dione (Example 21 Step 1) from
5-(3-fluorophenyl)-1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3-
,4-d]pyrimidine-7-carbaldehyde (Intermediate Nc) and
2-bromo-4-chlorothiazole (Intermediate Q, step 2);
[1534] 1H NMR (400 MHz, CDCl.sub.3) .delta. 12.16 (1H, s), 7.75
(1H, s), 7.69 (2H, d), 7.64 (2H, d), 7.48 (1H, mult), 7.29 (1H, d),
7.22 (1H, mult), 6.40 (1H, d), 3.48 (3H, s), 3.23 (3H, s)
[1535] LC-MS Rt 1.18 mins [M+H].sup.+ 421.2 m/z (Method
2minLowpHv03)
Step 2:
7-(1-(4-Chlorothiazol-2-yl)but-3-en-1-yl)-5-(3-fluorophenyl)-1,3-d-
imethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1536] Boron trifluoride THF complex (0.205 mL, 1.861 mmol) was
added to a solution of
7-((4-chlorothiazol-2-yl)(hydroxy)methyl)-5-(3-fluorophenyl)-1,3-dimethyl-
-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (870 mg, 2.067 mmol)
and allyltri-n-butyltin (0.641 mL, 2.067 mmol) were dissolved in
THF (15 mL). The mixture was stirred at room temp. for 20 mins. The
reaction mixture was quenched with saturated NaHCO.sub.3(aq) and
extracted with EtOAc. The organic extracts were washed with 1M
KF(aq) (1.times.), brine (1.times.), dried over sodium sulfate and
evaporated under vacuum. Purification by chromatography on silica,
eluting with 10-60% EtOAc/hexane, afforded the title compound.
[1537] LC-MS Rt 1.42 mins [M+H].sup.+ 445.3 (Method
2minLowpHv03)
[1538] 1H NMR (400 MHz, CDCl.sub.3) .delta. 9.92 (1H, s), 7.66 (1H,
dt), 7.60 (1H, d), 7.44 (1H, mult), 7.10 (1H, td), 7.05 (1H, s),
5.72 (1H, mult), 5.17-5.09 (2H, mult), 4.85 (1H, t), 3.68 (3H, s),
3.42 (3H, s), 2.92 (2H, mult).
Step 3:
7-(1-(4-Chlorothiazol-2-yl)-3,4-dihydroxybutyl)-5-(3-fluorophenyl)-
-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1539] Osmium tetroxide (2.5% w.t., 2.032 mL, 0.162 mmol) was added
to a solution of
7-(1-(4-chlorothiazol-2-yl)but-3-en-1-yl)-5-(3-fluorophenyl)-1,3-dimethyl-
-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (720 mg, 1.618 mmol)
and N-methyl morpholine oxide (284 mg, 2.427 mmol) in acetonitrile
(20 mL,) and water (2 mL). The mixture was stirred at room
temperature for 105 mins. The reaction was quenched with saturated
aqueous sodium metabisulfite solution and extracted with chloroform
(3.times.). The combined organic extracts were passed through a
hydrophobic frit and evaporated under vacuum to afford the title
compound as a mixture of diastereomers.
[1540] LC-MS Rt 1.12 mins [M+H].sup.+ 479.3 (Method
2minLowpHv03)
Step 4:
7-(4-((tert-Butyldimethylsilyl)oxy)-1-(4-chlorothiazol-2-yl)-3-hyd-
roxybutyl)-5-(3-fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4-
(3H,6H)-dione
[1541] tert-Butyldimethylsilyl chloride (309 mg, 2.047 mmol) was
added to a solution of
7-(1-(4-chlorothiazol-2-yl)-3,4-dihydroxybutyl)-5-(3-fluorophenyl)-1,3-di-
methyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (817 mg, 1.706
mmol), imidazole (232 mg, 3.41 mmol) and DMAP (20.84 mg, 0.171
mmol) in DMF (15 mL). The mixture was stirred at room temperature
for 50 mins. A further portion of tert-butyldimethylsilyl chloride
(100 mg, 0.662 mmol) was added and the mixture stirred for a
further 1 hour. The reaction mixture was diluted with EtOAc and
washed with 0.1M HCl(aq) (1.times.) and brine (3.times.). The
organic phase was dried over sodium sulfate and evaporated under
vacuum. Purification by chromatography on silica, eluting with
0-60% EtOAc/hexane, afforded the title compound as a mixture of
diastereomers.
[1542] LC-MS Rt 1.68 mins [M+H].sup.+ 593.5 (Method
2minLowpHv03)
Step 5:
1-((tert-Butyldimethylsilyl)oxy)-4-(4-chlorothiazol-2-yl)-4-(5-(3--
fluorophenyl)-1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]p-
yrimidin-7-yl)butan-2-yl methanesulfonate
[1543] Methanesulfonic anhydride (347 mg, 1.989 mmol) was added to
a solution of
7-(4-((tert-butyldimethylsilyl)oxy)-1-(4-chlorothiazol-2-yl)-3-hydroxybut-
yl)-5-(3-fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-
-dione (590 mg, 0.995 mmol) and triethylamine (0.416 mL, 2.98 mmol)
in DCM (30 mL). The mixture was stirred at room temperature 20
mins. The reaction was quenched with saturated NaHCO.sub.3(aq) and
extracted with DCM (3.times.). The combined organic extracts were
passed through a hydrophobic frit and evaporated under vacuum to
afford the title compound as a crude material which was used
directly.
Step 6:
7-(((tert-Butyldimethylsilyl)oxy)methyl)-9-(4-chlorothiazol-2-yl)--
5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine--
2,4(3H,7H)-dione
[1544] Sodium hydride (127 mg, 3.18 mmol) was added to a solution
of
1-((tert-butyldimethylsilyl)oxy)-4-(4-chlorothiazol-2-yl)-4-(5-(3-fluorop-
henyl)-1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidi-
n-7-yl)butan-2-yl methanesulfonate (712 mg, 1.061 mmol) in THF (30
mL). The mixture was stirred at room temperature for 35 mins. The
reaction was quenched with water and extracted with DCM (3.times.).
The combined organic extracts were passed through a hydrophobic
frit and evaporated under vacuum. Purification by chromatography on
silica, eluting with 0-100% EtOAc/hexane afforded the title
compound as a mixture of diastereomers.
[1545] LC-MS Rt 1.77 mins [M+H].sup.+ 575.4 (Method
2minLowpHv03)
Step 7: Diastereoisomeric mixture of
9-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-7-(hydroxymethyl)-1,3-dimethy-
l-8,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1546] Trifluoroacetic acid (2 mL, 26.0 mmol) was added to a
solution of 7-(((tert-butyldimethylsilyl)
oxy)methyl)-9-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9--
dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione (570 mg,
0.991 mmol) in methanol (20 mL) and water (3 mL). The mixture
stirred at room temperature 20 mins. Further portions of water (1
mL) and trifluoroacetic acid (1 mL) were added and the mixture
stirred for a further 3 hours. The reaction was quenched with
saturated NaHCO.sub.3(aq) and extracted with chloroform (3.times.).
The combined organic extracts were passed through a hydrophobic
frit and evaporated under vacuum. Purification by chromatography on
silica, eluting with 0-100% EtOAc/hexane afforded the title
compounds as a mixture of diastereomers.
[1547] Resolution of the diastereomers was carried out by serial
SFC chromatographic resolution under the following conditions;
[1548] Column: Chiralpak AD-H 250.times.10 mm 5 um
[1549] Eluent: 30% MeOH/60% CO2
[1550] Flow: 10 mL/min
[1551] Detection: UV @ 225 nm
[1552] followed by the following conditions;
[1553] Column: Chiralcel OJ-H 250.times.10 mm 5 um
[1554] Eluent: 30% MeOH/60% CO2
[1555] Flow: 10 mL/min
[1556] Detection: UV @ 225 nm
Example 27a
Diastereomer
(7R,9R)-9-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-7-(hydroxymethyl)-1,3-
-dimethyl-8,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1557] Rt=5.24 min
[1558] LC-MS Rt 1.12 mins [M+H].sup.+ 461.4 (Method
2minLowpHv03)
[1559] 1H NMR (400 MHz, CDCl3) .delta. 7.46 (1H, mult), 7.37 (1H,
mult), 7.30 (1H, mult), 7.15 (1H, dt), 7.06 (1H, s), 5.07 (1H, dd),
4.82 (1H, mult), 3.52-3.44 (2H, mult), 3.38 (1H, mult), 3.38 (3H,
s), 3.34 (3H, s), 2.83 (1H, dt)
Example 27b
Diastereomer
(7S,9R)-9-(4-chlorothiazol-2-yl)-5-(3-fluorophenyl)-7-(hydroxymethyl)-1,3-
-dimethyl-8,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1560] Rt=4.30 min
[1561] 1H NMR (400 MHz, CDCl3) .delta. 7.46 (1H, mult), 7.39 (1H,
mult), 7.31 (1H, mult), 7.16 (1H, td), 7.09 (1H, s), 5.11 (1H, dd),
4.86 (1H, mult), 3.52 (1H, dd), 3.47 (1H, dd), 3.37 (3H, s), 3.27
(3H, s), 3.22 (1H, mult), 2.88 (1H, mult)
[1562] LC-MS Rt 1.15 mins [M+H].sup.+ 461.4 (Method
2minLowpHv03)
[1563] The compounds of the following listed examples were prepared
by a similar method to that of Example 27a and 27b from the
appropriate starting material in step 1 (described hereinafter) and
appropriate halo compound. The resulting diastereomeric mixtures
were separated by SFC chromatographic under the listed
conditions.
Example 27.1a and 27.1b
(7R,9R)-9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-5-phenyl-8-
,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(7S,9R)-9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-5-phenyl-8-
,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(7R,9S)-9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-5-phenyl-8-
,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(7S,9S)-9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-5-phenyl-8-
,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00154##
[1565] Separation conditions: Serial separation
[1566] First separation:
[1567] Column: Chiralcel OJ-H 250.times.10 mm, 5 um @ 35 C
[1568] Eluent: 40% Methanol/60% CO2
[1569] Flow: 10 ml/min
[1570] Detection: UV @ 220 nm
[1571] Second separation:
[1572] Column: Chiralcel OD-H 500.times.10 mm, 5 um @ 35 C
[1573] Eluent: 30% Methanol/60% CO2
[1574] Flow: 10 ml/min
[1575] Detection: UV @ 220 nm
Example 27.1a
Diastereomer 1 of
9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-5-phenyl-8,9-dihy-
dro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1576] Rt=13.03 mins
[1577] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.58-7.52 (2H, mult),
7.50-7.39 (3H, mult), 7.04 (1H, s), 5.05 (1H, mult), 4.79 (1H,
mult), 3.49-3.36 (2H, mult), 3.33 (3H, s), 3.30 (3H, s), 2.81 (1H,
mult), 2.36 (1H, mult)
[1578] LC-MS Rt 1.09 mins [M+H].sup.+ 443.4 (Method
2minLowpHv03)
Example 27.1b
Diastereomer 2 of
9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-5-phenyl-8,9-dihy-
dro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1579] Rt=14.56 mins
[1580] LC-MS Rt 1.12 mins [M+H].sup.+ 443.3 (Method
2minLowpHv03)
[1581] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.61-7.55 (2H, mult),
7.51-7.41 (3H, mult), 7.09 (1H, s), 5.09 (1H, dd), 4.84 (1H, mult),
3.50-3.38 (2H, mult), 3.34 (3H, s), 3.26 (3H, s), 3.21 (1H, mult),
2.84 (1H, mult)
Example 27.2a and 27.2b
(7R,9R)-7-(hydroxymethyl)-1,3-dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8-
,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(7R,9R)-7-(hydroxymethyl)-1,3-dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8-
,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(7R,9R)-7-(hydroxymethyl)-1,3-dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8-
,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione or
(7R,9R)-7-(hydroxymethyl)-1,3-dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8-
,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00155##
[1583] Separation conditions:
[1584] Column: Chiralcel OJ-H 250.times.10 mm, 5 um @ 35 deg C.
[1585] Mobile phase: 35% Methanol/65% CO2
[1586] Flow: 10 ml/min
[1587] Detection: UV @ 220 nm
Example 27.2a
Diastereomer 1 of
7-(hydroxymethyl)-1,3-dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihy-
dro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1588] Rt=4.78 mins
[1589] LC-MS Rt 1.04 mins [M+H].sup.+ 423.4 (Method
2minLowpHv03)
[1590] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.58 (2H, d), 7.50-7.38
(3H, mult), 6.81 (1H, s), 5.01 (1H, d), 4.80 (1H, mult), 4.27 (1H,
mult), 3.55-3.34 (3H, mult), 3.32 (3H, s), 3.31 (3H, s), 2.69 (1H,
mult), 2.39 (3H, s).
Example 27.2b
Diastereomer 2 of
7-(hydroxymethyl)-1,3-dimethyl-9-(4-methylthiazol-2-yl)-5-phenyl-8,9-dihy-
dro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1591] Rt=4.49 mins
[1592] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.59 (2H, d), 7.51-7.41
(3H, mult), 6.85 (1H, s), 5.10 (1H, dd), 4.86 (1H, mult), 3.76 (1H,
mult), 3.51, 3.37 (2H, mult), 3.34 (3H, s), 3.22 (3H, s), 3.18 (1H,
mult), 2.84 (1H, mult), 2.47 (3H, s).
[1593] LC-MS Rt 1.06 mins [M+H].sup.+ 423.5 (Method
2minLowpHv03)
Example 27.3a and 27.3b
3-((7R,9R)-9-(4-Chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-2,4-dio-
xo-2,3,4,7,8,9-hexahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
or
3-((7R,9R)-9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-2,4-dio-
xo-2,3,4,7,8,9-hexahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
or
3-((7R,9R)-9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-2,4-dio-
xo-2,3,4,7,8,9-hexahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
or
3-((7R,9R)-9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-2,4-dio-
xo-2,3,4,7,8,9-hexahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
##STR00156##
[1595] Separation conditions:
[1596] Column: Chiralcel OJ-H 250.times.10 mm, 5 um @ 35 deg C.
[1597] Mobile phase: 35% Methanol/65% CO2
[1598] Flow: 10 ml/min
[1599] Detection: UV @ 220 nm
Example 27.3a
Diastereomer 1 of
3-(9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3,-
4,7,8,9-hexahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
[1600] Rt=4.81 mins
[1601] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.82-7.74 (2H, mult),
7.62 (1H, d), 7.51 (1H, t), 6.99 (1H, s), 4.98 (1H, d), 4.72 (1H,
mult), 3.46-3.33 (2H, mult), 3.27 (1H, mult, obscured), 3.25 (3H,
s, distinct), 3.25 (3H, s, distinct) 2.73 (1H, d), 2.46 (1H, br
s)
[1602] LC-MS Rt 1.07 mins [M+H].sup.+ 468.3 (Method
2minLowpHv03)
Example 27.3b
Diastereomer 2 of
3-(9-(4-chlorothiazol-2-yl)-7-(hydroxymethyl)-1,3-dimethyl-2,4-dioxo-2,3,-
4,7,8,9-hexahydro-1H-pyrimido[4,5-a]pyrrolizin-5-yl)benzonitrile
[1603] Rt=5.03 mins
[1604] 1H NMR (400 MHz, CDCl3) .delta. 7.89-7.83 (2H, mult), 7.70
(1H, d), 7.59 (1H, t), 7.11 (1H, s), 5.13 (1H, dd), 4.87 (1H,
mult), 3.50 (1H, dd), 3.40 (1H, dd), 3.31 (3H, s), 3.22 (1H, mult,
obscured) 3.22 (3H, s), 2.87 (1H, mult), 2.16 (1H, br s).
[1605] LC-MS Rt 1.10 mins [M+H].sup.+ 468.4 (Method
2minLowpHv03)
Example 28a and 28b
(8R,10R)-8-((1H-Imidazol-1-yl)methyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-
-5-(4-methylthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c]-
[1,4]thiazine-2,4(3H,10H)-dione or
(8R,10R)-8-((1H-imidazol-1-yl)methyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-
-5-(4-methylthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c]-
[1,4]thiazine-2,4(3H,10H)-dione or
(8R,10R)-8-((1H-imidazol-1-yl)methyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-
-5-(4-methylthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c]-
[1,4]thiazine-2,4(3H,10H)-dione or
(8R,10R)-8-((1H-imidazol-1-yl)methyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-
-5-(4-methylthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c]-
[1,4]thiazine-2,4(3H,10H)-dione
##STR00157## ##STR00158##
[1607] Sodium imidazolide (86 mg, 0.960 mmol) was added to a
solution of racemic
10-(5-chlorofuran-2-yl)-8-(hydroxymethyl)-1,3-dimethyl-5-(4-methy-
lthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazi-
ne-2,4(3H,10H)-dione (115 mg, 0.240 mmol) (Example 17) in DCM (24
ml) at 0.degree. C. The mixture was stirred for 30 minutes, then
trifluoromethanesulfonic anhydride (0.049 ml, 0.288 mmol) was added
dropwise. The mixture was warmed to room temperature and stirred
for 3 hours. The mixture was diluted with DCM (10 ml) washed with
water (30 ml) and brine (30 ml). The organic layer was passed
through a hydrophobic frit and evaporated under vacuum.
Purification by chromatography on silica, eluting with 0-20%
methanol/DCM, afford the title compound as a mixture of
diastereomers.
[1608] LC-MS Rt 0.90 min [M+H].sup.+ 529.3 (Method
2minLowpHv03)
[1609] The mixture of diastereomers was separated by SFC
chromatographic resolution under this following conditions to
afford the title compounds.
[1610] Column: Chiralpak IF 250.times.10 mm, 5 um @ 35 deg C.
[1611] Mobile phase: 50% methanol+0.1% DEA/50% CO2
[1612] Flow: 10 ml/min
[1613] Detection: UV @ 220 nm
Example 28a
Diastereomer 1 of
8-((1H-imidazol-1-yl)methyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-me-
thylthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thi-
azine-2,4(3H,10H)-dione
[1614] Rt=12.05 mins
[1615] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.32 (d, 1H), 7.26 (s,
1H), 7.00 (s, 1H), 6.73 (s, 1H), 6.19 (dd, 1H), 6.13 (d, 1H), 5.82
(s, 1H), 5.59 (dd, 1H), 4.37 (dd, 1H), 3.96 (dt, 1H), 3.85 (m, 2H),
3.69 (s, 3H), 3.46 (s, 3H), 2.47 (s, 3H)
[1616] LC-MS Rt 0.93 min [M+H].sup.+ 529.5 (Method
2minLowpHv03)
Example 28b
Diastereomer 2 of
8-((1H-imidazol-1-yl)methyl)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-me-
thylthiazol-2-yl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thi-
azine-2,4(3H,10H)-dione
[1617] Rt=7.40 mins
[1618] LC-MS Rt 0.90 min [M+H].sup.+ 529.4 (Method
2minLowpHv03)
[1619] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.33 (dd, 2H), 7.15 (d,
1H), 7.00 (s, 1H), 6.72 (s, 1H), 6.19 (dt, 1H), 6.13 (d, 1H), 5.81
(d, 1H), 5.59 (dd, 1H), 4.36 (dd, 1H), 3.98 (m, 1H), 3.85 (m, 2H),
3.69 (s, 3H), 3.46 (s, 3H), 2.47 (d, 3H)
Example 29
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8-methylene-8,9-d-
ihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
##STR00159##
[1621] Step 1: Dimethyl
2-((4-chlorothiazol-2-yl)(5-(3-fluorophenyl)-1,3-dimethyl-2,4-dioxo-2,3,4-
,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-7-yl)methyl)malonate
NaHMDS solution (1M in THF, 7.13 mL, 7.13 mmol) was added slowly to
a mixture of
7-((4-chlorothiazol-2-yl)(hydroxy)methyl)-5-(3-fluorophenyl)-1,3-dimethyl-
-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (Example 27, step 1)
(1 g, 2.376 mmol), dimethyl malonate (0.819 mL, 7.13 mmol) and
boron trifluoride THF complex (0.236 mL, 2.139 mmol) in THF (30
mL). The mixture was stirred at room temperature for 15 mins. The
reaction was quenched with saturated NaHCO.sub.3(aq) and extracted
with DCM (3.times.). The combined organic extracts were passed
through a hydrophobic frit and evaporated under vacuum.
Purification by chromatography on silica, eluting with 20-40%
EtOAc/hexane, afforded the title compound.
[1622] LC-MS Rt 1.37 mins [M+H].sup.+ 535.4 (Method
2minLowpHv03)
Step 2:
7-(1-(4-Chlorothiazol-2-yl)-3-hydroxy-2-(hydroxymethyl)propyl)-5-(-
3-fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1623] Sodium borohydride (190 mg, 5.02 mmol) was added to a
suspension of dimethyl
2-((4-chlorothiazol-2-yl)(5-(3-fluorophenyl)-1,3-dimethyl-2,4-di-
oxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-7-yl)methyl)malonate
(895 mg, 1.673 mmol) and lithium chloride (213 mg, 5.02 mmol) in
methanol (20 mL). The mixture was stirred at room temperature. for
30 mins. Further portions of lithium chloride (213 mg, 5.02 mmol)
and sodium borohydride (190 mg, 5.02 mmol) were added to allow the
reaction to run to completion. The reaction was quenched with
saturated NaHCO.sub.3(aq) and extracted with chloroform (3.times.).
The combined organic extracts were passed through a hydrophobic
frit and evaporated under vacuum to afford the title compound.
[1624] LC-MS Rt 1.18 mins [M+H].sup.+ 479.5 (Method
2minLowpHv03)
Step 3:
2-((4-Chlorothiazol-2-yl)(5-(3-fluorophenyl)-1,3-dimethyl-2,4-diox-
o-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-7-yl)methyl)allyl
methanesulfonate
[1625] Methanesulfonic anhydride (1699 mg, 9.75 mmol) was added to
a solution of
7-(1-(4-chlorothiazol-2-yl)-3-hydroxy-2-(hydroxymethyl)propyl)-5-(3-fluor-
ophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
(934 mg, 1.950 mmol) and triethylamine (1.631 mL, 11.70 mmol) in
DCM (20 mL). The mixture was stirred at room temperature for 16
hours. Further portions of triethylamine (1.631 mL, 11.70 mmol) and
methanesulfonic anhydride (1699 mg, 9.75 mmol) were added as
necessary to allow the reaction to run to completion. The reaction
was quenched with saturated NaHCO.sub.3(aq) and extracted with DCM
(3.times.). The combined organic extracts were passed through a
hydrophobic frit and evaporated under vacuum. Purification by
chromatography on silica, eluting with 20-100% EtOAc/hexane,
afforded the title compound.
[1626] LC-MS Rt 1.27 mins [M+H].sup.+ 539.5 (Method
2minLowpHv03)
Step 4:
9-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8-methyle-
ne-8,9-dihydro-1H-pyrimido[4,5-a]pyrrolizine-2,4(3H,7H)-dione
[1627] Sodium hydride (60% wt. in mineral oil, 31.2 mg, 0.779 mmol)
was added to a solution of
2-((4-chlorothiazol-2-yl)(5-(3-fluorophenyl)-1,3-dimethyl-2,4-dioxo-2,3,4-
,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-7-yl)methyl)allyl
methanesulfonate (140 mg, 0.260 mmol) in THF (8 mL). The mixture
was stirred at room temperature for 2 hours. A further portion of
sodium hydride (31.2 mg, 0.779 mmol) was added to allow the
reaction to run to completion. The reaction was quenched with water
and extracted with DCM (3.times.). The combined organic extracts
were passed through a hydrophobic frit and evaporated under vacuum.
Purification by chromatography on silica, eluting with 20-100%
EtOAc/hexane, afforded the title compound.
[1628] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.45 (1H, mult), 7.39
(1H, mult), 7.34-7.25 (2H, mult), 7.13 (1H, mult), 7.10 (1H, s),
4.65 (1H, dd), 3.97 (1H, d), 3.39 (3H, s), 3.23 (3H, s), 2.64 (1H,
mult), 2.61 (1H, mult)
[1629] LC-MS Rt 1.35 mins [M+H].sup.+ 443.4 (Method
2minLowpHv03)
Example 30a, 30b, 30c and 30d
(8R,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-(-
(2-oxooxazolidin-3-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,-
1-c][1,4]thiazine-2,4(3H,10H)-dione or
(8S,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-(-
(2-oxooxazolidin-3-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,-
1-c][1,4]thiazine-2,4(3H,10H)-dione or
(8S,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-(-
(2-oxooxazolidin-3-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,-
1-c][1,4]thiazine-2,4(3H,10H)-dione or
(8R,10S)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-(-
(2-oxooxazolidin-3-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,-
1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00160## ##STR00161##
[1630] Step 1: 2-Chloroethyl
((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dioxo-
-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-8-
-yl)methyl)carbamate
[1631] The title compound was prepared in a similar manner to
Example 26 step 1, replacing methanesulfonyl chloride with
2-chloroethyl chloroformate.
[1632] LC-MS Rt 1.36 min [M+H]+ 584.3 (Method 2minLowpHv03)
Step 2:
10-(5-Chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((-
2-oxooxazolidin-3-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-
-c][1,4]thiazine-2,4(3H,10H)-dione
[1633] Sodium hydride (60% wt in mineral oil, 6.69 mg, 0.167 mmol)
was added to a solution of 2-chloroethyl
((10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-2,4-dioxo-
-2,3,4,7,8,10-hexahydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4]thiazin-8-
-yl)methyl)carbamate (48.9 mg, 0.084 mmol) in THF (4 mL). The
mixture was heated at reflux for 18 hours. A further portion of
sodium hydride (60% wt in mineral oil, 33.4 mg, 0.836 mmol) was
added and the mixture heated at reflux for a further 2 days. The
mixture was cooled to room temperature, quenched with water (20 ml)
and extracted with ethyl acetate (2.times.30 ml). The combined
organic extracts were washed with brine, passed through a
hydrophobic frit and evaporated under vacuum. Purification by
mass-directed HPLC under the following conditions afforded the
title compound as a mixture of diastereomers.
[1634] Column: XSelect CSH Prep C18 column, 30.times.100 mm, 5
um.
[1635] Mobile phase: A=0.1% FA in water, B=0.1% FA in MeCN
[1636] Gradient:
[1637] 0.0-0.5 min: 30% B 30 mL/min
[1638] 0.5-1.0 min: 30% B 30-50 mL/min
[1639] 1.0-7.2 min: 30-70% B, 7.2-7.3 min: 70-98% B, 7.3-9.4 min:
98% B 9.4-9.5 30% B 50 mL/min
[1640] The mixture of diastereomers was separated by SFC
chromatographic resolution under the listed conditions to afford
the title compounds as single diastereomers.
[1641] Column: Chiralpak IA, 250.times.10 mm, 5 um @ 35.degree.
C.
[1642] Mobile phase: 40% Methanol/60% CO2
[1643] Flow: 10 ml/min
[1644] Detection: UV @ 220 nm
Example 30a
Diastereomer 1 of
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxoox-
azolidin-3-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazine-2,4(3H,10H)-dione
[1645] Rt=7.02 mins
[1646] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.13 (s, 1H), 6.16 (d,
1H), 6.13 (d, 1H), 5.79 (s, 1H), 5.34 (dd, 1H), 4.47 (dd, 1H), 4.26
(m, 2H), 3.92 (q, 1H), 3.66 (s, 3H), 3.61 (m, 1H), 3.44 (s, 3H),
3.37 (m, 2H), 3.10 (dd, 1H), 2.53 (s, 3H)
[1647] LC-MS Rt 1.24 min [M+H].sup.+ 548.3 (Method
2minLowpHv03)
Example 30b
Diastereomer 2 of
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxoox-
azolidin-3-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazine-2,4(3H,10H)-dione
[1648] Rt=9.45 mins
[1649] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.13 (s, 1H), 6.22 (t,
1H), 6.15 (d, 1H), 5.90 (dd, 1H), 5.80 (s, 1H), 4.39 (td, 2H), 3.77
(m, 4H), 3.67 (s, 3H), 3.63 (m, 1H), 3.48 (dd, 1H), 3.44 (s, 3H),
2.52 (s, 3H)
[1650] LC-MS Rt 1.24 min [M+H].sup.+ 548.4 (Method
2minLowpHv03)
Example 30c
Diastereomer 3 of
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxoox-
azolidin-3-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazine-2,4(3H,10H)-dione
[1651] Rt=7.91 mins
[1652] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.13 (s, 1H), 6.22 (t,
1H), 6.15 (d, 1H), 5.90 (dd, 1H), 5.80 (s, 1H), 4.39 (td, 2H), 3.77
(m, 4H), 3.67 (s, 3H), 3.63 (m, 1H), 3.48 (dd, 1H), 3.44 (s, 3H),
2.52 (s, 3H)
[1653] LC-MS Rt 1.24 min [M+H].sup.+ 548.4 (Method
2minLowpHv03)
Example 30d
Diastereomer 4 of
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxoox-
azolidin-3-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazine-2,4(3H,10H)-dione
[1654] Rt=6.25 mins
[1655] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.13 (s, 1H), 6.16 (d,
1H), 6.13 (d, 1H), 5.79 (s, 1H), 5.34 (dd, 1H), 4.47 (dd, 1H), 4.26
(m, 2H), 3.92 (q, 1H), 3.66 (s, 3H), 3.61 (m, 1H), 3.44 (s, 3H),
3.37 (m, 2H), 3.10 (dd, 1H), 2.53 (s, 3H)
[1656] LC-MS Rt 1.24 min [M+H].sup.+ 520.3 (Method
2minLowpHv03)
[1657] The following examples were prepared in a similar manner to
Example 30a-d, replacing 2-chloroethyl chloroformate in step 1 with
4-bromobutyryl chloride. The mixture of diastereomers was separated
by SFC chromatographic resolution under the listed conditions to
afford the title compounds as single diastereomers.
Example 30.1a, 30.1b, 30.1c and 30.1d
(8R,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-(-
(2-oxopyrrolidin-1-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,-
1-c][1,4]thiazine-2,4(3H,10H)-dione or
(8R,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-(-
(2-oxopyrrolidin-1-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,-
1-c][1,4]thiazine-2,4(3H,10H)-dione or
(8R,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-(-
(2-oxopyrrolidin-1-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,-
1-c][1,4]thiazine-2,4(3H,10H)-dione or
(8R,10R)-10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-(-
(2-oxopyrrolidin-1-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,-
1-c][1,4]thiazine-2,4(3H,10H)-dione
##STR00162## ##STR00163##
[1659] Separation conditions:
[1660] Column: Chiralcel OD-H 250.times.10 mm, 5 um @ 35.degree.
C.
[1661] Mobile phase: 40% Methanol/60% CO2
[1662] Flow: 10 ml/min
[1663] Detection: UV @ 220 nm
Example 30.1a
Diastereomer 1 of
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxopy-
rrolidin-1-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazine-2,4(3H,10H)-dione
[1664] Rt=5.25 mins
[1665] 1H NMR (400 MHz, CDCl3) .delta. 7.13 (s, 1H), 6.12 (s, 2H),
5.76 (s, 1H), 5.08 (dd, 1H), 4.51 (dd, 1H), 3.87 (m, 1H), 3.65 (s,
3H), 3.43 (s, 3H), 3.39 (m, 2H), 3.22 (dd, 1H), 3.12 (dd, 1H), 2.53
(s, 3H), 2.33 (t, 2H), 1.97 (m, 2H)
[1666] LC-MS Rt 1.25 min [M+H].sup.+ 546.4 (Method
2minLowpHv03)
Example 30.1d
Diastereomer 4 of
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxopy-
rrolidin-1-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazine-2,4(3H,10H)-dione
[1667] Rt=7.69 mins
[1668] LC-MS Rt 1.25 min [M+H].sup.+ 546.4 (Method
2minLowpHv03)
[1669] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.16 (s, 1H), 6.29 (d,
1H), 6.15 (d, 1H), 5.78 (s, 1H), 5.60 (d, 1H), 3.85 (d, 2H), 3.68
(s, 3H), 3.68 (m, 1H), 3.60 (dd, 1H), 3.51 (dd, 1H), 3.44 (m, 1H),
3.43 (s, 3H), 2.55 (s, 3H), 2.39 (t, 2H), 2.09 (q, 2H)
[1670] Diastereomers 2 and 3 were further separated under the
following conditions to afford single diasteromers
[1671] Column: LUX A2, 250.times.10 mm, 5 um @35.degree. C.
[1672] Mobile phase: 50% Methanol/50% CO2
[1673] Flow: 10 ml/min
[1674] Detection: UV @ 220 nm
Example 30.1b
Diastereomer 2 of
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxopy-
rrolidin-1-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazine-2,4(3H,10H)-dione
[1675] Rt=14.80 mins
[1676] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.16 (s, 1H), 6.29 (d,
1H), 6.15 (d, 1H), 5.78 (s, 1H), 5.60 (d, 1H), 3.85 (d, 2H), 3.68
(s, 3H), 3.68 (m, 1H), 3.60 (dd, 1H), 3.51 (dd, 1H), 3.44 (m, 1H),
3.43 (s, 3H), 2.55 (s, 3H), 2.39 (t, 2H), 2.09 (q, 2H)
[1677] LC-MS Rt 1.25 min [M+H].sup.+ 546.4 (Method
2minLowpHv03)
Example 30.1c
Diastereomer 3 of
10-(5-chlorofuran-2-yl)-1,3-dimethyl-5-(4-methylthiazol-2-yl)-8-((2-oxopy-
rrolidin-1-yl)methyl)-7,8-dihydro-1H-pyrimido[4',5':3,4]pyrrolo[2,1-c][1,4-
]thiazine-2,4(3H,10H)-dione
[1678] Rt=10.38 mins
[1679] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.13 (s, 1H), 6.12 (s,
2H), 5.76 (s, 1H), 5.08 (dd, 1H), 4.51 (dd, 1H), 3.87 (m, 1H), 3.65
(s, 3H), 3.43 (s, 3H), 3.39 (m, 2H), 3.22 (dd, 1H), 3.12 (dd, 1H),
2.53 (s, 3H), 2.33 (t, 2H), 1.97 (m, 2H)
[1680] LC-MS Rt 1.25 min [M+H].sup.+ 546.4 (Method
2minLowpHv03)
Example 31
(R)-3-(11-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9,10,11--
octahydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepin-5-yl)benzonitrile
or
(S)-3-(11-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9,10,11--
octahydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepin-5-yl)benzonitrile
##STR00164##
[1681] Step 1: Ethyl
5-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)pentanoate
[1682] The title compound was prepared from
3-(1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-
-yl)benzonitrile (Example 9, step 4) and ethyl 5-bromopentanoate
(0.712 mL, 4.50 mmol) by an analogous method to Example 9 step
5;
[1683] LC-MS: Rt 1.18 mins; MS 409.6 m/z [M+H] Method
2minLowpHv03
[1684] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.79-7.69 (3H, mult),
7.62 (1H, t), 6.47 (1H, s), 4.14 (2H, q), 3.93 (2H, t), 3.45 (3H,
s), 3.36 (3H, s), 2.25 (2H, t), 1.74 (2H, mult), 1.52 (2H, mult),
1.26 (3H, t).
Step 2:
5-(5-(3-Cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo-
[3,4-d]pyrimidin-6(2H)-yl)-N-methoxy-N-methylpentanamide
[1685] The title compound was prepared from ethyl
5-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)pentanoate by a method similar to Example 13,
step 2;
[1686] LC-MS: Rt 1.06 mins; MS 424.3 m/z [M+H] Method
2minLowpHv03
[1687] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.77-7.70 (3H, mult),
7.61 (1H, mult), 6.49 (1H, s), 3.92 (2H, t), 3.66 (3H, s), 3.43
(3H, s), 3.36 (3H, s), 3.17 (3H, s), 2.38 (2H, t), 1.75 (2H, mult),
1.54 (2H, mult).
Step 3:
3-(6-(5-(4-Chlorothiazol-2-yl)-5-oxopentyl)-1,3-dimethyl-2,4-dioxo-
-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
[1688] The title compound was prepared from
5-(5-(3-cyanophenyl)-1,3-dimethyl-2,4-dioxo-3,4-dihydro-1H-pyrrolo[3,4-d]-
pyrimidin-6(2H)-yl)-N-methoxy-N-methylpentanamide and
2-bromo-4-chlorothiazole by a method similar to Example 13, step
3;
[1689] LC-MS: Rt 1.30 mins; MS 482.2 m/z [M+H] Method
2minLowpHv03
[1690] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.78-7.70 (3H, mult),
7.62 (1H, t), 7.49 (1H, s), 6.49 (1H, s), 3.97 (2H, t), 3.44 (3H,
s), 3.36 (3H, s), 3.06 (2H, t), 1.79 (2H, mult), 1.64 (2H,
mult).
Step 4:
3-(6-(5-(4-Chlorothiazol-2-yl)-5-hydroxypentyl)-1,3-dimethyl-2,4-d-
ioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
[1691] The title compound was prepared from
3-(6-(5-(4-Chlorothiazol-2-yl)-5-oxopentyl)-1,3-dimethyl-2,4-dioxo-2,3,4,-
6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile by a
method similar to Example 13, step 4;
Step 5:
3-(11-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9,10-
,11-octahydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepin-5-yl)benzonitrile
[1692] The title compound was prepared from
3-(6-(5-(4-chlorothiazol-2-yl)-5-hydroxypentyl)-1,3-dimethyl-2,4-dioxo-2,-
3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile by a
method similar to Example 13, step 5;
[1693] LC-MS: Rt 1.34 mins; MS 466.4 m/z [M+H] Method
2minLowpHv04.
Step 6: (R) or
(S)-3-(11-(4-Chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9,10,11-
-octahydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepin-5-yl)benzonitrile
[1694] Chiral separation of racemic
3-(11-(4-chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9,10,11-oct-
ahydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepin-5-yl)benzonitrile
by Supercritical Fluid Chromatography was carried out using the
following conditions to afford the title compound;
[1695] Column: Chiralcel AD-H 250.times.10 mm, 5 um @ 35 C
[1696] Mobile phase: 15% Methanol/85% CO2
[1697] Flow: 10 ml/min
[1698] Detection: UV @ 220 nm
Enantiomer 1 of
3-(11-(4-chlorothiazol-2-yl)-1,3-dimethyl-2,4-dioxo-2,3,4,7,8,9,10,11-oct-
ahydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepin-5-yl)benzonitrile
[1699] SFC Retention time=26.26 mins
[1700] LC-MS: Rt 1.36 mins; MS 466.5 m/z [M+H] Method
2minLowpHv03
[1701] 1H NMR (400 MHz, CDCl3) .delta. 7.70-7.49 (4H, mult), 7.01
(1H, s), 5.23 (1H, dd), 3.93 (1H, dd), 3.54 (3H, s), 3.39 (1H, dd),
3.26 (3H, s), 2.90 (1H, mult), 2.02-1.89 (5H, mult)
[1702] The second enantiomer was isolated at SFC Rt=32.07 min
Example 32
(R)-11-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9,10,11-te-
trahydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione
or
(S)-11-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9,10,11-te-
trahydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione
##STR00165##
[1703] Step 1:
5-(3-Fluorophenyl)-1,3-dimethyl-6-((2-(trimethylsilyl)ethoxy)methyl)-1H-p-
yrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1704] The title compound was prepared from
1,3-dimethyl-2,4-dioxo-6-((2-(trimethylsilyl)ethoxy)methyl)-2,3,4,6-tetra-
hydro-1H-pyrrolo[3,4-d]pyrimidin-5-ylboronic acid (Example 9, step
2) and 1-bromo-3-fluorobenzene by a similar method to Example 9
step 3;
[1705] LCMS; Rt 1.54 mins; MS m/z 404.4 [M+H]+; 2minlowpHV03.
Step 2:
5-(3-Fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H-
,6H)-dione
[1706] The title compound was prepared from
5-(3-Fluorophenyl)-1,3-dimethyl-6-((2-(trimethylsilyl)ethoxy)methyl)-1H-p-
yrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione by a similar method to
Example 9, step 4;
[1707] LC-MS: Rt 1.05 mins; MS m/z 274.3 [M+H]+; Method
2minLowpHv03
Step 3-9: (R)- or
(S)-11-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9,10,11-t-
etrahydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione
[1708] The title compound was prepared by a similar method to
Example 31 steps 1-6;
Enantiomer 1 of
11-(4-Chlorothiazol-2-yl)-5-(3-fluorophenyl)-1,3-dimethyl-8,9,10,11-tetra-
hydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione
[1709] SFC Rt=4.38 mins SFC Chiralcel OJ-H 250.times.10 mm, 5 um @
35 deg C., 35% Methanol/65% CO2, 10 ml/min, Detection: UV @ 220 nm,
>99% e.e.
[1710] LC-MS Rt 1.45 mins; MS 459.4 m/z [M+H] Method
2minLowpHv03
[1711] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.46 (1H, mult),
7.23-7.10 (3H, mult), 7.09 (1H, s), 5.30 (1H, mult), 4.11 (1H,
mult), 3.64 (3H, s), 3.43 (1H, mult), 3.37 (3H, s), 3.01 (1H,
mult), 2.11-1.88 (5H, mult)
Example 33
1,3-Dimethyl-11-(4-methylthiazol-2-yl)-5-phenyl-8,9,10,11-tetrahydro-1H-py-
rimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione
##STR00166##
[1712] Step 1: Ethyl
5-(1,3-dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-
-6(2H)-yl)pentanoate
[1713]
5-Benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
(Intermediate C) (1.38 g, 4.09 mmol), ethyl 5-aminovalerate
hydrochloride (1.109 g, 6.14 mmol) and triethylamine (1.711 mL,
12.28 mmol) were combined in ethanol (20 mL) and the mixture heated
at reflux for 40 mins. The reaction mixture was cooled to room
temperature and evaporated under vacuum. The residue was
partitioned between water and DCM and the phases separated. The
organic phase was passed through a hydrophobic frit and evaporated
onto silica. The silica was deposited onto a 25 g silica cartridge
and the system eluted with 20% EtOAc/hexane, 40% EtOAc/hexane, 60%
EtOAc/hexane and 80% EtOAc/hexane. The product fractions were
combined and concentrated under reduced pressure to afford the
title compound;
[1714] LC-MS: Rt 1.29 mins; MS 384.3 m/z [M+H] Method
2minLowpHv03
[1715] 1H NMR (400 MHz, CDCl3) .delta. 7.54-7.37 (5H, mult), 6.41
(1H, br s), 4.12 (2H, q), 3.93 (2H, t), 3.44 (3H, s), 3.36 (3H, s),
2.22 (2H, t), 1.74 (2H, mult), 1.53 (2H, mult), 1.26 (3H, t).
Step 2-5:
1,3-Dimethyl-11-(4-methylthiazol-2-yl)-5-phenyl-8,9,10,11-tetrah-
ydro-1H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione
[1716] The title compound was prepared from ethyl
5-(1,3-dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidin-
-6(2H)-yl)pentanoate by a similar method to Example 31 steps
2-5;
[1717] LC-MS: Rt 1.34 mins; MS 421.2 m/z [M+H] Method
2minLowpHv03
[1718] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.55-7.34 (5H, mult),
6.95 (1H, s), 5.45 (1H, mult), 4.18 (1H, mult), 3.63 (3H, s), 3.36
(3H, s), 3.25 (1H, mult), 2.61 (3H, s), 2.19-1.79 (5H, mult).
Example 33a
(R)-1,3-Dimethyl-11-(4-methylthiazol-2-yl)-5-phenyl-8,9,10,11-tetrahydro-1-
H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione or
(S)-1,3-Dimethyl-11-(4-methylthiazol-2-yl)-5-phenyl-8,9,10,11-tetrahydro-1-
H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione
##STR00167##
[1720] Chiral separation of racemic
1,3-dimethyl-11-(4-methylthiazol-2-yl)-5-phenyl-8,9,10,11-tetrahydro-1H-p-
yrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione (Example
33) by Supercritical Fluid Chromatography was carried out using the
following conditions to afford the title compound;
[1721] SFC Chiralcel OJ-H 250 mm.times.10 mm.times.5 .mu.m, Eluent
35% IPA (containing 0.1% DEA)/65% CO2, 10 ml/min @ 35.degree. C.,
detection @ 220 nm,
Enantiomer 1 of
1,3-Dimethyl-11-(4-methylthiazol-2-yl)-5-phenyl-8,9,10,11-tetrahydro-1H-p-
yrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione
[1722] SFC Retention time=3.96 min, >99% ee
[1723] LC-MS: Rt 1.39 mins; MS 421.3 m/z [M+H] Method
2minLowpHv03
[1724] 1H NMR (av81851, 400 MHz, CDCl.sub.3) .delta. 7.52-7.37 (5H,
mult), 6.88 (1H, s), 5.37 (1H, dd), 4.14 (1H, dd), 3.63 (3H, s),
3.45 (1H, dd), 3.36 (3H, s), 3.10 (1H, mult), 2.54 (3H, s), 2.08
(1H, mult), 1.91 (2H, mult), 1.57 (2H, mult).
[1725] The second entantiomer was isolated at SFR Rt=6.31 mins
Example 34
(R)-11-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9,10,11-tetrahydro-1-
H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione or
(S)-11-(4-Chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9,10,11-tetrahydro-1-
H-pyrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione
##STR00168##
[1727] The title compound was prepared by a similar method to
Example 33a by replacing 2-iodo-4-methylthiazole (Example 33, step
3) with 2-bromo-4-chlorothiazole (Intermediate Q, step 2);
Enantiomer 1 of
11-(4-chlorothiazol-2-yl)-1,3-dimethyl-5-phenyl-8,9,10,11-tetrahydro-1H-p-
yrimido[4',5':3,4]pyrrolo[1,2-a]azepine-2,4(3H,7H)-dione
[1728] LC-MS: Rt 1.45 mins; MS 441.3 m/z [M+H] Method
2minLowpHv03
[1729] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.54-7.34 (5H, mult),
7.08 (1H, s), 5.03 (1H, mult), 4.14 (1H, mult), 3.63 (3H, s), 3.37
(1H, mult), 3.36 (3H, s), 3.00 (1H, mult), 2.12-1.85 (5H,
mult).
[1730] SFC Chiralcel OJ-H 250.times.10 mm, 5 um @35 deg C., 45%
Methanol+0.1% v/v DEA/55% CO2, 10 ml/min, Detection: UV @ 220 nm,
Rt 4.79 min, >99% e.e.
Preparation of Intermediates
Intermediate A
6-(2-Mercapto-ethyl)-1,3-dimethyl-5-phenyl-1,6-dihydro-pyrrolo[3,4-d]pyrim-
idine-2,4-dione
##STR00169##
[1731] Step 1:
1,3-Dimethyl-5-phenyl-6-(2-tritylsulfanyl-ethyl)-1,6-dihydro-pyrrolo[3,4--
d]pyrimidine-2,4-dione
[1732] Triethylamine (0.211 ml, 1.513 mmol) was added to a solution
of
5-Benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione(Interme-
diate C) (510 mg, 1.513 mmol) and 2-tritylsulfanyl-ethylamine
(Intermediate D) (483 mg, 1.513 mmol) in EtOH (5 ml) and the
mixture heated at 100.degree. C. under microwave irradiation for 2
hours. The reaction mixture was diluted with DCM (10 mL) and was
washed with water (3.times.10 mL). The organic phase was passed
through a hydrophobic frit and the solvent was evaporated under
reduced pressure. The title product was obtained as a pale yellow
solid and used without further purification.
[1733] LC-MS Rt 1.37 mins [M+H].sup.+ 558.3 (Method 2minLowpH)
Step 2:
6-(2-Mercapto-ethyl)-1,3-dimethyl-5-phenyl-1,6-dihydro-pyrrolo[3,4-
-d]pyrimidine-2,4-dione
[1734] TFA (5.975 ml, 78 mmol) was added to a solution of
1,3-dimethyl-5-phenyl-6-(2-tritylsulfanyl-ethyl)-1,6-dihydro-pyrrolo[3,4--
d]pyrimidine-2,4-dione (step 1) (865 mg, 1.551 mmol) and
triethylsilane (2.477 ml, 15.51 mmol) in DCM (20 ml) and the
mixture stirred at RT for 3 hours. The solvent was evaporated under
reduced pressure. The residue was partitioned between DCM (10 mL)
and sat. NaHCO.sub.3 (10 mL) and the phases separated. The organic
phase was dried over magnesium sulfate and the solvent evaporated
under reduced pressure. The residue was triturated with diethyl
ether and the solid collected by filtration. The title compound was
obtained as a white solid.
[1735] .sup.1H NMR (400 MHz, DMSO-d6) .delta. 7.45 (5H, s), 7.01
(1H, s), 4.02 (2H, t), 3.31 (3H, s), 3.15 (3H, s), 2.80 (2H, q),
2.33 (5H, t).
[1736] LC-MS Rt 0.95 mins; [M+H].sup.+ 316.5 (Method 2minLowpH)
Intermediate B
(S)-6-(1-(Dimethylamino)-3-mercaptopropan-2-yl)-1,3-dimethyl-5-phenyl-1H-p-
yrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
##STR00170##
[1737] Step 1: (S)-2-Amino-3-(tritylthio)propan-1-ol
[1738] D-Cysteine (2.36 g, 19.48 mmol) was suspended in THF (40 mL)
and cooled to 0.degree. C. Borane-tetrahydrofuran complex (1 M in
THF, 78 mL, 78 mmol) was added over 20 mins and the mixture was
stirred at RT for 6 hours. The reaction was quenched with DMF (7
mL) and stirred for 25 mins. Trityl chloride (5.97 g, 21.43 mmol)
was added and the mixture stirred for a further 16 hours. The
reaction mixture was evaporated under vacuum. The residue was
partitioned between DCM and water and the phases separated. The
organic phase was passed through a hydrophobic frit and evaporated
under vacuum. Purification by chromatography on silica, eluting
with 50% EtOAc/hexane, EtOAc, 10% MeOH/EtOAc and 20% MeOH/EtOAc
gave the title compound.
[1739] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.49-7.17 (1%, mult),
3.55 (1H, mult), 3.40 (1H, mult), 2.74 (1H, mult), 2.49 (2H,
mult).
Step 2:
(S)-6-(1-Hydroxy-3-(tritylthio)propan-2-yl)-1,3-dimethyl-5-phenyl--
1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1740] (S)-2-Amino-3-(tritylthio)propan-1-ol (step 1) (1.58 g, 4.52
mmol), triethylamine (1.454 mL, 10.43 mmol) and
5-benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
(Intermediate C) (1.173 g, 3.48 mmol) were combined in EtOH (60 mL)
and the mixture heated at reflux for 1 hour. The reaction mixture
was cooled to RT and evaporated under vacuum. The residue was
partitioned between DCM and water and the phases separated. The
organic phase was passed through a hydrophobic frit and evaporated
under vacuum to give the title compound which was used without
further purification.
[1741] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.56-7.17 (20H,
mult), 6.37 (1H, s), 4.17 (1H, mult), 3.67 (2H, mult), 3.43 (3H,
s), 3.35 (3H, s), 2.70 (1H, mult), 2.51 (1H, mult).
[1742] LC-MS Rt=1.40 min [M+H]+ 588.6 (Method 2minLowpHv01).
Step 3:
(S)-2-(1,3-Dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4--
d]pyrimidin-6(2H)-yl)-3-(tritylthio)propyl methanesulfonate
[1743]
(S)-6-(1-Hydroxy-3-(tritylthio)propan-2-yl)-1,3-dimethyl-5-phenyl-1-
H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (step 2) (2.5 g, 4.25
mmol) was dissolved in DCM (80 mL) and triethylamine (1.186 mL,
8.51 mmol) was added followed by methanesulfonyl chloride (0.398
mL, 5.10 mmol). The mixture was stirred at room temperature for 2
hours, further methanesulfonyl chloride was added and the mixture
stirred for 50 mins. The reaction mixture was quenched with sat.
NaHCO.sub.3(aq) and extracted with DCM. The organic phase was
passed through a hydrophobic frit and evaporated under vacuum.
Purification by chromatography on silica, eluting with 20-100%
EtOAc in iso-hexane gave the title compound.
[1744] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.50 (4H, mult),
7.39 (2H, mult), 7.35-7.22 (9H, mult), 6.30 (1H, s), 4.33 (1H,
mult), 4.17 (1H, mult), 4.09 (1H, mult), 3.43 (3H, s), 3.34 (3H,
s), 2.82 (3H, s), 2.73 (1H, mult), 2.49 (1H, mult).
[1745] LC-MS Rt=1.45 min [M+H]+ 666.3 (Method 2minLowpHv01).
Step 4:
6-((S)-1-Dimethylaminomethyl-2-tritylsulfanyl-ethyl)-1,3-dimethyl--
5-phenyl-1,6-dihydro-pyrrolo[3,4-d]pyrimidine-2,4-dione
[1746]
(S)-2-(1,3-Dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d-
]pyrimidin-6(2H)-yl)-3-(tritylthio)propyl methanesulfonate (step
3)(1.7 g, 2.55 mmol) was dissolved in DMF (12 mL) and dimethylamine
(40% wt. aqueous solution) (1.455 mL, 11.49 mmol) was added. The
mixture was heated under microwave irradiation at 120.degree. C.
for 15 h. The reaction mixture was diluted with EtOAc and washed
with brine (4.times.). The organic phase was dried over sodium
sulphate and evaporated under vacuum. The residue was redissolved
in DCM and evaporated onto silica. Purification by chromatography
on silica, eluting with 20-100% EtOAc in iso-hexane gave the title
compound.
[1747] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.51 (5H, mult),
6.42 (1H, s), 4.35 (1H, mult), 3.45 (3H, s), 3.36 (3H, s), 2.95
(2H, mult), 2.64 (2H, mult), 1.57 (6H, mult).
[1748] LC-MS: Rt 1.07 mins; MS 615.7 m/z [M+H] Method
2minLowpHv01
Step 5:
(S)-6-(1-(Dimethylamino)-3-mercaptopropan-2-yl)-1,3-dimethyl-5-phe-
nyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
##STR00171##
[1750]
6-((S)-1-Dimethylaminomethyl-2-tritylsulfanyl-ethyl)-1,3-dimethyl-5-
-phenyl-1,6-dihydro-pyrrolo[3,4-d]pyrimidine-2,4-dione (step 4)
(1.3 g, 2.115 mmol) was dissolved in DCM (50 mL) and TFA (5 mL,
64.9 mmol) and triethylsilane (0.675 mL, 4.23 mmol) were added. The
mixture was stirred at RT for 1 hour 20mins. The reaction mixture
was rigorously evaporated under vacuum. Purification by
chromatography on silica, eluting with 20-100% EtOAc/hexane, 10%
MeOH/EtOAc and 10% (7N NH3 in MeOH)/EtOAc gave the title
compound.
[1751] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.51 (5H, mult),
6.42 (1H, s), 4.35 (1H, mult), 3.45 (3H, s), 3.36 (3H, s), 2.95
(2H, mult), 2.64 (2H, mult), 1.57 (6H, mult)
[1752] LC-MS: Rt 0.64 mins; MS 373.5 m/z [M+H] Method
2minLowpHv01
Intermediate C
5-Benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
##STR00172##
[1753] Step 1: 1,3,6-Trimethylpyrimidine-2,4(1H,3H)-dione
[1754] To a stirred suspension of N,N'-dimethylurea (commercial)
(72.1 g, 819 mmol) and DMAP (100 g, 819 mmol) in pyridine (300 mL)
under N.sub.2 was added dropwise with stirring acetic anhydride
(255 mL, 2701 mmol). On complete addition the reaction mixture was
allowed to stir at RT for 3 hours. After this time, the volatiles
were removed under reduced pressure to afford a viscous orange
pyridine solution, which was seeded with product. The mixture was
stored at 0-4.degree. C. for 7 days. The resulting crystalline
solid was then filtered under reduced pressure, washed with diethyl
ether and dried to afford the title compound as colourless
crystals. The mother liquors were further purified by
chromatography on silica eluting with 30-50% EtOAc in iso-hexane.
The resulting solid was diluted with diethyl ether (500 mL) and was
stored at 0-4.degree. C. overnight. The resulting crystals were
filtered off and washed with iso-hexane, then dried to afford the
title compound as colourless crystals and combined. The title
product was obtained as white crystals.
[1755] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.58 (1H, s), 3.38
(3H, s), 3.30 (3H, s), 2.22 (3H, s);
[1756] LC-MS Rt=0.55 min [M+H]+ 155.4 (Method: 2minLC_v003).
Step 2: 5-Benzoyl-1,3,6-trimethylpyrimidine-2,4(1H,3H)-dione
[1757] 1,3,6-Trimethylpyrimidine-2,4(1H,3H)-dione (step 1) (5 g,
32.4 mmol) and chlorobenzene (50 mL) were charged to a 3 necked
flask, and the vessel evacuated with nitrogen. Benzoyl chloride
(commercial) (11.2 mL, 97 mmol) was added followed by zinc (II)
chloride (5 g, 36.7 mmol) in one portion and the reaction was then
heated to 110.degree. C. for 16 h. The reaction was cooled to RT
then poured into water (100 mL) and EtOAc (100 mL). The layers were
separated and the aqueous extracted with EtOAc (2.times.150 mL).
The combined organics were washed with water (100 mL) and dried
(Na.sub.2SO.sub.4), filtered under reduced pressure and evaporated
to leave an orange solid. The solid was washed with hexane followed
by hot diethyl ether, affording the product as an off white solid.
The mother liquors were further purified by chromatography on
silica, eluting with 10%-100% EtOAc/hexane. The title product was
obtained as a pale orange solid;
[1758] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.89 (2H, dd),
7.60 (1H, t), 7.48 (2H, t), 3.52 (3H, s), 3.38 (3H, s), 2.26 (3H,
s);
[1759] LCMS Rt=0.80 min [M+H]+ 259 (Method 2minLC_v003).
Step 3:
5-Benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
[1760] Bromine (0.497 mL, 9.68 mmol) was added to a solution of
5-benzoyl-1,3,6-trimethyl pyrimidine-2,4(1H,3H)-dione (step 2) (2.5
g, 9.68 mmol) in chloroform (50 mL) with stirring under nitrogen.
The mixture was then heated to 55.degree. C. for 2 h. A further
portion of bromine (0.25 mL, 0.5 equiv) was then added and heating
continued at 55.degree. C. for a further 30 min. The mixture was
then cooled to RT, diluted with CHCl.sub.3 (50 mL) and poured into
saturated sodium thiosulfate solution (150 mL). The layers were
separated and the aqueous phase was extracted with CHCl.sub.3 (50
mL). The combined organics were washed with water (50 mL), brine
(50 mL), dried (Na.sub.2SO.sub.4) and filtered under reduced
pressure. The solvent was then evaporated under reduced pressure to
afford a pale yellow solid. This solid was dissolved in hot EtOAc
(50 mL) and slowly evaporated until crystallisation was observed.
The resulting white solid was collected by reduced pressure
filtration and washed with cold EtOAc (10 mL) and dried in air. The
compound was further dried under vacuum at 50.degree. C. for 2 h.
The title product was isolated as white micro needles.
[1761] .sup.1H NMR (400 MHz, DMSO-d.sub.6) .delta. 7.90 (2H, d),
7.67 (1H, t), 7.52 (2H, t), 4.40 (2H, s), 3.50 (3H, s), 3.17 (3H,
s);
[1762] LC-MS Rt=2.75 min [M+H].sup.+ 337/339 (Method
10minLC_v003).
Intermediate D
2-Tritylsulfanyl-ethylamine
##STR00173##
[1764] Cysteamine (commercial) (900 mg, 11.67 mmol) was added to a
stirred solution of triphenylmethanol (commercial) (3.037 g, 11.67
mmol) in TFA (10 ml) and the mixture stirred at RT for 3 hours. The
mixture was then concentrated under reduced pressure and the
residue partitioned between DCM (20 mL) and water (20 mL). The
mixture was basified by the slow addition of solid K.sub.2CO.sub.3.
The aqueous phase was extracted with DCM (2.times.20 mL), the
combined organic phases passed through a hydrophobic frit and the
solvent evaporated under reduced pressure. The material was used
without further purification.
Intermediate E
(S)-6-(2-Mercaptopropyl)-1,3-dimethyl-5-phenyl-1H-pyrrolo[3,4-d]pyrimidine-
-2,4(3H,6H)-dione
##STR00174##
[1765] Step 1:
(R)-6-(2-Hydroxypropyl)-1,3-dimethyl-5-phenyl-1H-pyrrolo[3,4-d]pyrimidine-
-2,4(3H,6H)-dione
[1766]
5-Benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
(Intermediate C) (1 g 2.97 mmol) was added to a microwave vial
(10-25 mL) equipped with a stirrer bar. To the vial was added
ethanol (10 ml) followed by triethylamine (0.41 ml, 2.97 mmol) and
(R)-1-aminopropan-2-ol (commercial). (223 mg, 2.97 mmol) The vial
was heated to 100.degree. C. in a microwave reactor for 1 h. The
ethanol was removed in vacuo to yield a solid. The solid was
dissolved in DCM and water added. The DCM water mixture was
separated and the aqueous extracted with DCM (.times.2). The
organics were combined, washed with water and passed through a
hydrophobic frit. The solvent was removed in vacuo to yield the
title compound as a pale yellow solid.
[1767] LC-MS: Rt 0.86 mins; MS m/z 314 [M+H]+; Method
2minLowpHv01
Step 2:
(R)-1-(1,3-Dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4--
d]pyrimidin-6(2H)-yl)propan-2-yl methanesulfonate
[1768] To a solution of
(R)-6-(2-hydroxypropyl)-1,3-dimethyl-5-phenyl-1H-pyrrolo[3,4-d]pyrimidine-
-2,4(3H,6H)-dione (step 1) (925 mg, 2.95 mmol) in DCE (16 ml) was
added triethylamine (2.05 ml, 14.8 mmol), DMAP (36 mg, 0.29 mmol)
and methanesulfonyl chloride (1.15 ml, 14.8 mmol) at 0.degree. C.
and reaction stirred for 30 min. Solid K.sub.2CO.sub.3 was added to
the reaction mixture and the resulting mixture was diluted with
water and extracted with DCM (3.times.20 ml) The organics were
passed through a hydrophobic frit and concentrated in vacuo to
yield a yellow oil. The material was used without further
purification.
[1769] LC-MS Rt 1.04 mins; MS m/z 392 [M+H]+; (Method
2minLowpHv01)
Step 3:
(S)-1,3-Dimethyl-5-phenyl-6-(2-(tritylthio)propyl)-1H-pyrrolo[3,4--
d]pyrimidine-2,4(3H,6H)-dione
[1770] To a stirred solution of triphenylmethylthiol (0.93 mg, 3.37
mmol) in dry THF (50 ml) was added sodium hydride (225 mg, 5.62
mmol) at 0.degree. C. and the solution stirred for 1 hour at RT. To
the resulting yellow solution was added crude
(R)-1-(1,3-dimethyl-2,4-dioxo-5-phenyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrim-
idin-6(2H)-yl)propan-2-yl methanesulfonate (step 2) (1.1 mg, 2.81
mmol) as a solution in dry DMF (3 ml) at 0.degree. C. The solution
was stirred overnight at RT. The reaction was poured into dilute
HCl and extracted with EtOAc, The organic layer was successively
washed with water and dried over Na.sub.2SO.sub.4. The solvent was
removed in vacuo to yield a yellow oil/semi-solid. The oil was
dissolved in a minimal volume of DCM and purified via ISCO column
chromatography, 24 g, liquid load, 20-35% iso-hexane:EtOAc, product
eluting at 30% affording 12 fractions. The corresponding fractions
were reduced in vacuo to yield an oil of title compound. The
material was used without further purification.
[1771] LC-MS Rt 1.55 mins; MS m/z 572 [M+H]+ (Method
2minLowpHv01)
Step 4:
(S)-6-(2-Mercaptopropyl)-1,3-dimethyl-5-phenyl-1H-pyrrolo[3,4-d]py-
rimidine-2,4(3H,6H)-dione
[1772] To crude
(S)-1,3-dimethyl-5-phenyl-6-(2-(tritylthio)propyl)-1H-pyrrolo[3,4-d]pyrim-
idine-2,4(3H,6H)-dione (step 3)(1.35 mg, 1.06 mmol) in DCM (30 ml)
was added TFA (4.09 ml, 53.1 mmol) and triethylsilane (1.7 ml, 10.6
mmol) giving a yellow solution. The mixture was stirred at RT
overnight. The reaction mixture was evaporated under vacuum. The
residue was partitioned between DCM/water and the phases separated.
The organic phase was passed through a hydrophobic frit and
evaporated under vacuum. The solid was dissolved in a minimal
volume of DCM and purified via ISCO column chromatography, 40 g
silica, liquid load, 0-60% iso-hexane:EtOAc, product eluting at 50%
affording 7 pure fractions. Corresponding fractions were reduced in
vacuo to yield the title compound as a white solid.
[1773] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.52-7.43 (5H, m); 6.44
(1H, s); 4.01 (2H, d); 3.44 (3H, s); 3.36 (3H, s); 3.12 (1H,
sextet); 1.43 (1H, d); 1.16 (3H, d).
[1774] LCMS Rt 1.00 mins; MS m/z 330 [M+H]+; (Method
2minLowpHv01)
Intermediate F
1,3-Dimethyl-2,4-dioxo-6-(2-(tritylthio)ethyl)-2,3,4,6-tetrahydro-1H-pyrro-
lo[3,4-d]pyrimidin-5-ylboronic acid
##STR00175##
[1775] Step 1 (2-Chloroethyl)(trityl)sulfane
[1776] Synthesised by a modified Method of Trujillo, D. A.;
McMahon, W. A.; Lyle, R. E. J. Org. Chem., 1987, 52, 2932-2933
[1777] (Chloromethanetriyl)tribenzene (100 g, 359 mmol) was
dissolved in dichloromethane (500 mL) under nitrogen. Ethylene
sulfide (25 mL, 420 mmol) was charged and the reaction aged at room
temperature for 22 h. The solvent was then evaporated under reduced
pressure to afford a pale yellow solid. The solid was dissolved in
hot (60.degree. C.) toluene (150 mL) and hot filtered under
gravity, then cooled slowly to room temperature. The resultant
white crystalline solid was collected by reduced pressure
filtration and washed with toluene (2.times.100 mL) and iso-hexanes
(2.times.100 mL), dried under vacuum at 50.degree. C. for 16 h. The
title compound was isolated as a white crystalline solid;
[1778] LCMS Rt 1.57 mins; MS m/z 243.4 [M+H].sup.+; Method
2minLowpHv01;
[1779] 1H NMR (400 MHz, DMSO-d6) .delta. 7.38-7.32 (12H, m),
7.30-7.24 (3H, m), 3.21 (2H, t), 2.55 (2H, t).
Step 2:
1,3-Dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1780] 1,3-Dimethylpyrimidine-2,4(1H,3H)-dione (commercially
available, 40 g, 285 mmol) and toluenesulfonylmethyl isocyanide
(commercially available, 84 g, 428 mmol) were dissolved in 2-MeTHF
(1000 mL) under nitrogen at 30.degree. C. and held for 5 mins. The
vessel was cooled to 0.degree. C. (internal). A solution of KOtBu
(commercially available, 64.1 g, 571 mmol) in 2-MeTHF (500 mL was
then added to the solution via a dropping funnel over 0.5 h, such
that the internal temperature remained below 5.degree. C. On
addition of the KOtBu solution, an orange precipitate formed. An
additional portion of 2-MeTHF (455 mL) was then charged via
dropping funnel over 5 mins. After 40 mins, the dark orange
suspension was quenched with sat NH.sub.4Cl solution (2.times.400
mL) charged via dropping funnel over 15 mins. The suspension was
diluted with EtOAc (1 L), the layers were separated and the aqueous
extracted with EtOAc (3.times.1 L). The combined organics were
dried (MgSO.sub.4) and filtered under reduced pressure. The solvent
was removed under reduced pressure to afford a brown semi-solid.
The solids were suspended in MeOH (300 mL), sonicated and stirred
at room temperature for 15 mins, filtered by reduced pressure
filtration and washed with MeOH (100 mL). The solid was dried under
vacuum at 50.degree. C. for 24 h. The title compound was obtained
as a pale brown amorphous solid;
[1781] 1H NMR (400 MHz, DMSO-d6) .delta. 11.81 (1H br s), 7.43 (1H,
d); 6.74 (1H, d); 3.29 (3H, s); 3.20 (3H, s)
[1782] LC-MS: Rt 0.59 mins; not ionised; Method 2minLowpHv01
[1783] A second crop of material could be obtained by evaporation
of the MeOH to afford a red oil. Trituration with MeOH (50 mL),
filtration under reduced pressure and washing with MeOH (20 mL) and
drying under vacuum at 50.degree. C. for 16 h afforded the title
compound as a pale brown amorphous solid;
[1784] 1H NMR (400 MHz, DMSO-d6) .delta. 11.82 (1H, s), 7.43 (1H,
dd), 6.74 (1H, t), 3.29 (3H, s), 3.19 (3H, s).
Step 3:
1,3-Dimethyl-6-(2-(tritylthio)ethyl)-1H-pyrrolo[3,4-d]pyrimidine-2-
,4(3H,6H)-dione
[1785] 1,3-Dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione)
(step 2)(10 g, 55.8 mmol), (2-chloroethyl)(trityl)sulfane (step 1)
(20.81 g, 61.4 mmol), potassium iodide (1.853 g, 11.16 mmol) and
caesium carbonate (36.4 g, 112 mmol) were suspended in anhydrous
dimethylacetamide (250 mL), forming a dark orange suspension. The
vessel was evacuated and back-filled with N.sub.2 (.times.3) and
stirred under nitrogen at 50.degree. C. for 41.7 h. The reaction
was cooled to room temperature and decanted slowly into a stirring
water (800 mL) and EtAcO (200 mL) mixture. The layers were
separated and the aqueous extracted with EtOAc (4.times.100 mL).
The solids present where collected by reduced pressure filtration
(solid 1). The combined organics were washed with water
(4.times.100 mL), brine (2.times.100 mL), dried (Na.sub.2SO.sub.4)
and filtered under reduced pressure. The organic solvent was
evaporated under reduced pressure to afford a pale yellow solid,
which was suspended in Et.sub.2O (200 mL) and stirred at room
temperature for 10 mins; the pale tan solid was then collected by
reduced pressure filtration (solid 2). The solid collected from the
extraction mixture (solid 1) was washed with water (100 mL), EtOAc
(100 mL) and stirred in Et.sub.2O (100 mL) for 10 mins at room
temperature and collected by reduced pressure filtration. Both
solids were dried under vacuum at 50.degree. C. for 16 h. Both
crops were then blended to afford the title compound as a tan
solid;
[1786] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.46-7.39 (6H, m),
7.36-7.29 (6H, m), 7.29-7.22 (3H, m), 7.01 (1H, d), 6.03 (1H d),
3.48 (2H, t), 3.38 (3H, s), 3.33 (3H, s), 2.67 (2H, t).
[1787] LC-MS: Rt 1.38 mins; MS m/z 482.2 [M+H].sup.+; Method
2minLowpHv01
Step 4:
1,3-Dimethyl-2,4-dioxo-6-(2-(tritylthio)ethyl)-2,3,4,6-tetrahydro--
1H-pyrrolo[3,4-d]pyrimidin-5-ylboronic acid
[1788]
1,3-Dimethyl-6-(2-(tritylthio)ethyl)-1H-pyrrolo[3,4-d]pyrimidine-2,-
4(3H,6H)-dione (step 2)(3.6 g, 7.47 mmol) was dissolved in
anhydrous THF (67.3 mL) at room temperature in an oven dried flask.
The flask was evacuated and backfilled with nitrogen (.times.3) and
cooled to -78.degree. C. LDA (0.719 M, 16.63 mL, 11.96 mmol) was
added over 10 mins followed by triisopropyl borate (2.76 mL, 11.96
mmol) and the reaction aged for 2 hr at -78.degree. C. The reaction
was quenched with saturated NH.sub.4Cl solution (60 mL) added via
syringe under nitrogen and the mixture was warmed to room
temperature and stirred overnight under nitrogen. The mixture was
then diluted DCM (400 mL). The layers were separated and the
aqueous extracted with DCM (2.times.100 mL). The combined organics
were passed through a hydrophobic frit and the solvent was reduced
to .about.20 mL under reduced pressure and EtOAc (100 mL) added to
the resulting red oil. Reduction of solvent volume under reduced
pressure to .about.25 mL afforded precipitation of a white solid.
The solid was collected by reduced pressure filtration, washed with
EtOAc (10 mL) and dried under vacuum 50.degree. C. for 16 h to
afford the title compound;
[1789] LC-MS: Rt 1.41 mins; MS m/z 526.4 [M+H]+; Method
2minLowpHv01;
[1790] 1H NMR (400 MHz, DMSO-d6) .delta. 9.61 (2H, s), 7.36-7.29
(6H, m), 7.29-7.20 (9H, m), 6.79 (1H, s), 4.27 (2H, t), 3.28 (3H,
s), 3.27 (3H, s), 2.54 (2H, t).
Intermediate G
3-(6-(2-Mercaptoethyl)-1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrol-
o[3,4-d]pyrimidin-5-yl)benzonitrile
##STR00176##
[1791] Step 1:
3-(1,3-Dimethyl-2,4-dioxo-6-(2-(tritylthio)ethyl)-2,3,4,6-tetrahydro-1H-p-
yrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
[1792] A suspension comprising
1,3-dimethyl-2,4-dioxo-6-(2-(tritylthio)ethyl)-2,3,4,6-tetrahydro-1H-pyrr-
olo[3,4-d]pyrimidin-5-ylboronic acid (Intermediate F) (300 mg,
0.571 mmol), 3-bromobenzonitrile (104 mg, 0.571 mmol) and barium
hydroxide (196 mg, 1.142 mmol) in n-BuOAc (5710 .mu.L) was treated
with Pd-118 (18.61 mg, 0.029 mmol) and the mixture was heated to
80.degree. C. Water (247 .mu.L, 13.70 mmol) was added at 80.degree.
C. and the suspension was stirred rapidly for 90 minutes. After
cooling to RT, the mixture was allowed to stand at RT overnight.
The mixture was diluted with DCM (50 ml) and washed with water (50
ml). The layers were separated and the aqueous portion was
extracted with DCM (3.times.30 ml). The organics were combined and
dried over MgSO.sub.4, filtered and concentrated. Purification of
the crude product by trituration with EtOAc/iso-hexane afforded the
title compound;
[1793] LCMS: Rt 1.45 min, [M+H]+ 583.6. Method 2minLowpHv01.
Step 2:
3-(6-(2-Mercaptoethyl)-1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1-
H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
[1794]
3-(1,3-Dimethyl-2,4-dioxo-6-(2-(tritylthio)ethyl)-2,3,4,6-tetrahydr-
o-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile (step 1)(573 mg,
0.983 mmol) was dissolved in DCM (20 ml) to form a clear brown
solution and passed through 2.times.500 mg Isolute.RTM. Si-TMT
(silica bound equivalent of 2,4,6-trimercatotriazine) cartridges to
afford a colourless solution. The solution was placed under a
nitrogen atmosphere and whilst stirring rapidly, treated dropwise
with triethylsilane (1.571 ml, 9.83 mmol) followed TFA (3.79 ml,
49.2 mmol). After stirring at RT for 1 h, the reaction mixture was
concentrated under reduced pressure to yield a white solid. The
isolated crude product was triturated with diethyl ether and dried
in a vacuum oven at 50.degree. C. to afford the title compound;
[1795] LCMS: Rt 0.99 min, [M+H]+ 341.3. Method 2minLowpHv01.
Intermediate GA
3-Chloro-5-(1,3-dimethyl-2,4-dioxo-6-(2-(tritylthio)ethyl)-2,3,4,6-tetrahy-
dro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
##STR00177##
[1797] The title compound was prepared analogously to
3-(1,3-dimethyl-2,4-dioxo-6-(2-(tritylthio)
ethyl)-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile
(Intermediate GA step 1) by replacing bromobenzonitrile with
3-bromo-5-chlorobenzonitrile;
[1798] LC-MS: Rt 1.53 mins; MS m/z 617.3/619.3 [M+H]+; Method
2minLowpHv01
Intermediate GB
Ethyl
2-(1,3-dimethyl-2,4-dioxo-6-(2-(tritylthio)ethyl)-2,3,4,6-tetrahydro-
-1H-pyrrolo[3,4-d]pyrimidin-5-yl)thiazole-4-carboxylate
##STR00178##
[1800]
(1,3-dimethyl-2,4-dioxo-6-(2-(tritylthio)ethyl)-2,3,4,6-tetrahydro--
1H-pyrrolo[3,4-d]pyrimidin-5-yl)boronic acid (Intermediate F) (1001
mg, 1.906 mmol), ethyl 2-bromothiazole-4-carboxylate (409 mg, 1.732
mmol), sodium hydrogen carbonate (291 mg, 3.46 mmol) and Pd-118
(113 mg, 0.173 mmol) were suspended in n-butyl acetate (12.3 ml)
and heated to 80.degree. C. Water (3.08 ml) was then added and the
mixture heated at 80.degree. C. for 21 hours. The mixture was then
cooled to room temperature and diluted with water (100 ml) and DCM
(100 ml). The mixture was extracted with DCM (3.times.100 ml) and
the combined organic extracts were washed with water (50 ml) and
brine (50 ml). Silica-TMT (30 g) was added and the mixture stirred
at room temperature for 5 mins, then filtered. The residue was
rinsed with DCM (20 ml) and the filtrates evaporated under vacuum.
The resultant residue was rinsed with diethyl ether and filtered to
afford the title compound.
[1801] 1H NMR (400 MHz, DMSO-d6) .delta. 8.62 (1H, s), 7.30-7.24
(6H, m), 7.24-7.18 (9H, s), 6.85 (1H, s), 4.36 (2H, t), 4.32 (2H,
q), 3.31 (3H, s), 3.23 (3H, s), 2.57 (2H, t), 1.30 (3H, t)
[1802] LC-MS Rt 1.76 mins [M+H].sup.+ 637.5 (Method
2minLowpHv03)
Intermediate H
1,3-Diethyl-6-(2-mercaptoethyl)-5-phenyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3-
H,6H)-dione
##STR00179##
[1803] Step 1: 1,3-Diethyl-6-methylpyrimidine-2,4(1H,3H)-dione
[1804] 6-Methylpyrimidine-2,4(1H,3H)-dione (5.5 g, 43.6 mmol) was
suspended in water (80 ml) and cooled to 0.degree. C. NaOH (17.44
g, 436 mmol) was added slowly, forming a solution. Ethyl iodide
(14.10 ml, 174 mmol) was added and the mixture heated at 60.degree.
C. for 3 days. The reaction mixture was cooled to RT and diluted
with water. The mixture was extracted with chloroform (3.times.).
The basic extracts were evaporated under vacuum. The residue was
dissolved in DCM and evaporated onto silica. The silica was
deposited onto a 25 g silica cartridge and the system eluted
progressively with 10% EtOAc/isohexane, 20% EtOAc/isohexane, 40%
EtOAc/isohexane, 75% EtOAc/isohexane and EtOAc to yield the title
compound as a colourless oil;
[1805] 1H NMR (400 MHz, CDCl.sub.3) .delta. 5.59 (1H, s), 4.01 (2H,
q), 3.92 (2H, q), 2.26 (3H, s), 1.30 (3H, t), 1.24 (3H, t).
[1806] LC-MS Rt 0.72 mins; MS 183.5 m/z [M+H](Method
2minLowpHv01)
Step 2:
5-Benzoyl-1,3-diethyl-6-methylpyrimidine-2,4(1H,3H)-dione
[1807] 1,3-Diethyl-6-methylpyrimidine-2,4(1H,3H)-dione (step 1)(680
mg, 3.73 mmol), benzoyl chloride (1.299 mL, 11.20 mmol) and tin(IV)
chloride (1.314 mL, 11.20 mmol) were combined in chlorobenzene (30
mL, 295 mmol) and the mixture heated at reflux for 16 h. The
reaction mixture was cooled to RT and diluted with EtOAc. The
mixture was washed with 2M NaOH(aq) until basic. The organic phase
was washed with brine, dried over sodium sulphate and evaporated
under vacuum. The residue was dissolved in DCM and evaporated onto
silica. The silica was deposited onto a 10 g silica cartridge and
the system eluted with 10% EtOAc/hexane, 20% EtOAc/hexane, 40%
EtOAc/hexane and 50% EtOAc/hexane. Product elutes at 40%
EtOAc/hexane. Fractions were concentrated under vacuum to yield the
title compound as an off white solid;
[1808] 1H NMR (, 400 MHz, CDCl.sub.3) .delta. 7.89 (2H, d), 7.60
(1H, t), 7.48 (2H, t), 4.03 (2H, q), 4.02 (2H, q), 2.27 (3H, s),
1.37 (3H, t), 1.24 (3H, t).
[1809] LC-MS Rt 1.00 mins; MS 287.5 m/z [M+H](Method
2minLowpHv01)
Step 3:
5-Benzoyl-6-(bromomethyl)-1,3-diethylpyrimidine-2,4(1H,3H)-dione
[1810] 5-Benzoyl-1,3-diethyl-6-methylpyrimidine-2,4(1H,3H)-dione
(step 2)(800 mg, 2.79 mmol) was dissolved in chloroform (Volume: 60
mL) and bromine (0.288 mL, 5.59 mmol) was added. The mixture was
heated at 60.degree. C. for 1 h. A further portion of bromine
(0.288 mL, 5.59 mmol) was added. The mixture was heated for 30 mins
and further bromine (0.288 mL, 5.59 mmol) was added. The reaction
mixture was cooled to RT and washed with aqueous sodium
metabisulfite. The organic phase was separated, passed through a
hydrophobic frit and evaporated under vacuum. The residue was
dissolved in DCM and evaporated onto silica. The silica was
deposited onto a 10 g silica cartridge and the system eluted with
20% EtOAc/hexane, 30% EtOAc/hexane, and 40% EtOAc/hexane. Product
containing fractions were concentrated under vacuum. The resultant
oily residue was triturated with hexane, which was collected by
filtration. The title compound was collected as an off white
solid.
[1811] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.87 (2H, d), 7.62 (1H,
t), 7.49 (2H, t), 4.26 (2H, s), 4.16 (2H, q), 4.02 (2H, q), 1.43
(3H, t), 1.25 (3H, t).
Step 4:
1,3-Diethyl-5-phenyl-6-(2-(tritylthio)ethyl)-1H-pyrrolo[3,4-d]pyri-
midine-2,4(3H,6H)-dione
[1812]
5-Benzoyl-6-(bromomethyl)-1,3-diethylpyrimidine-2,4(1H,3H)-dione
(step 3) (850 mg, 2.327 mmol), 2-(tritylthio)ethanamine
(commercially available, 1115 mg, 3.49 mmol) and triethylamine
(0.973 mL, 6.98 mmol) were combined in EtOH (Volume: 20 mL) and the
mixture heated at reflux for 20 mins. The reaction mixture was
cooled to RT and evaporated under vacuum. The residue was
partitioned between DCM and water and extracted with DCM. The
organic phase was passed through a hydrophobic frit and evaporated
onto silica. The silica was deposited onto a 10 g silica cartridge
and purification by chromatography on silica eluting with 10-50%
EtOAc in iso-hexane afforded the title compound as a white
solid;
[1813] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.45 (4H, mult),
7.40-7.21 (16H, mult), 5.99 (1H, s), 4.01 (2H, mult), 3.88 (2H,
mult), 3.66 (2H, mult), 2.49 (2H, t), 1.34 (3H, mult), 1.19 (3H,
mult).
[1814] LC-MS Rt 1.59 mins; MS 586.3 m/z [M+H](Method
2minLowpHv02)
Step 5:
1,3-Diethyl-6-(2-mercaptoethyl)-5-phenyl-1H-pyrrolo[3,4-d]pyrimidi-
ne-2,4(3H,6H)-dione
[1815]
1,3-Diethyl-5-phenyl-6-(2-(tritylthio)ethyl)-1H-pyrrolo[3,4-d]pyrim-
idine-2,4(3H,6H)-dione (step 4) (1.45 g, 2.475 mmol) was dissolved
in DCM (50 mL) and treated with TFA (5 mL, 64.9 mmol) followed by
triethylsilane (0.791 mL, 4.95 mmol). The mixture was stirred at RT
for 16 h. The reaction mixture was basified by slow addition of
sat. NaHCO.sub.3(aq) and extracted with DCM. The organic phase was
passed through a hydrophobic frit and evaporated under vacuum. The
residue was dissolved in DCM, evaporated onto silica and the silica
deposited onto a 10 g silica cartridge. The system was eluted with
20%-60% EtOA in iso-hexane to yield the title compound as a pale
yellow oil.
[1816] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.53-7.44 (5H, mult),
6.46 (1H, s), 4.11 (2H, t), 4.03 (2H, q), 3.94 (2H, q), 2.74 (1H,
dt), 1.37 (3H, t), 1.21 (3H, t).
[1817] LC-MS Rt 1.19 mins; MS 344.6 m/z [M+H](Method
2minLowpHv01)
Intermediate I
1,3-Dimethyl-5-phenyl-7,8-dihydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dion-
e
##STR00180##
[1819]
5-Benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
(Intermediate C), (1.169 g, 3.47 mmol) in EtOH (20 mL) was treated
with 4,4-diethoxybutan-1-amine (0.659 mL, 3.81 mmol) and TEA (0.966
mL, 6.93 mmol) and the mixture heated at reflux for 50 min. The
reaction mixture was cooled to RT and evaporated under reduced
pressure. The residue was re-dissolved in THF (20 mL), 1M
hydrochloric acid (3.47 mL) was added and the mixture stirred at RT
for 1.5 h. The reaction mixture was diluted with EtOAc, washed with
water, brine and the organic phase was dried over sodium sulphate
and evaporated under reduced pressure. The residue was triturated
with DCM/hexane and the precipitate collected by filtration to give
the title product as a pale pink solid.
[1820] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.53-7.44 (m, 5H), 6.85
(dt, 1H), 5.84 (dt, 1H), 3.97 (t, 2H), 3.66 (s, 3H), 3.37 (s, 3H),
2.47 (m, 2H);
[1821] LC-MS Rt=1.11 min [M+H]+ 308 (Method 2minLC_v003).
Intermediate J
4-Methyl-2-tributylstannanyl-thiazole
##STR00181##
[1823] 4-Methylthiazole (0.5 ml, 5.50 mmol) was dissolved in
Et.sub.2O (10 ml) and cooled to -78.degree. C. Methyllithium (1.6 M
in hexanes, 5.15 ml, 8.24 mmol) was added slowly and the mixture
stirred at -78.degree. C. for 1 hour. Tri-n-butyltin chloride
(1.640 ml, 6.05 mmol) was then added slowly, the mixture stirred
briefly at -78.degree. C. then allowed to warm to RT and stirred
overnight. The reaction mixture was quenched with water and
extracted with Et.sub.2O. The organic phase was washed with brine,
dried over sodium sulphate and evaporated under vacuum to afford
the title product as a crude pale yellow oil. The material was used
without further purification.
Intermediate K
1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4,7,8-hexahydropyrimido[4,5-a]indoli-
zin-10-yl trifluoromethanesulfonate
##STR00182##
[1824] Step 1:
4-(1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4-tetrahydro-pyrrolo[3,4-d]pyrim-
idin-6-yl)-butyric acid ethyl ester
[1825]
5-Benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
(Intermediate C) (996 mg, 2.95 mmol), ethyl 4-aminobutanoate
hydrochloride (489 mg, 2.95 mmol) and triethylamine (0.823 ml, 5.91
mmol) were combined in EtOH (15 ml) and heated at reflux for 2.5
hours. The reaction mixture was cooled to RT and evaporated under
reduced pressure. The residue was dissolved in EtOAc and washed
with water (1.times.) and brine (1.times.). The organic phase was
dried over sodium sulphate and evaporated under reduced pressure to
afford the title compound as an off-white solid;
[1826] 1H NMR (400 MHz, CDCl3) .delta. 7.51-7.41 (m, 5H), 6.43 (br
s, 1H), 4.08 (q, 2H), 3.99 (t, 2H), 3.44 (s, 3H), 3.36 (s, 3H),
2.20 (t, 2H), 2.00 (m, 2H), 1.24 (t, 3H);
[1827] LC-MS Rt 1.11 min [M+H].sup.+ 370.5 (Method: 2minLowpH)
Step 2:
4-(1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4-tetrahydro-pyrrolo[3,4--
d]pyrimidin-6-yl)-butyric acid
[1828]
4-(1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4-tetrahydro-pyrrolo[3,4-d-
]pyrimidin-6-yl)-butyric acid ethyl ester (step 1)(1.01 g, 2.73
mmol) was dissolved in THF (10 ml) and water (10 ml) and lithium
hydroxide (0.655 g, 27.3 mmol) were added. The mixture was stirred
at 60.degree. C. for 5 hours. Further lithium hydroxide (0.655 g,
27.3 mmol) was added and the mixture stirred at 60.degree. C. for a
further 24 hours. The reaction mixture was cooled to RT and
concentrated under reduced pressure. The residue was acidified
using 1M HCl(aq) and extracted with chloroform (5.times.). The
combined organic phases were passed through a hydrophobic frit and
evaporated under reduced pressure. The title product was obtained
as a white solid.
[1829] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.52-7.41 (m, 5H), 6.42
(s, 1H), 4.00 (t, 2H), 3.43 (s, 3H), 3.35 (s, 3H), 2.25 (t, 2H),
2.00 (m, 2H);
[1830] LC-MS Rt 0.82 min [M+H].sup.+ 342.5 (Method: 2minLowpH)
Step 3:
1,3-Dimethyl-5-phenyl-8,9-dihydropyrimido[4,5-a]indolizine-2,4,10(-
1H,3H,7H)-trione
[1831]
4-(1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4-tetrahydro-pyrrolo[3,4-d-
]pyrimidin-6-yl)-butyric acid (step 2)(1.3 g, 3.81 mmol) and
polyphosphoric acid (10 g, 3.81 mmol) were combined and heated at
80.degree. C. for 1 hour. The reaction mixture was cooled to RT,
diluted with water and extracted with chloroform (4.times.). The
combined organic extracts were passed through a hydrophobic frit
and evaporated under reduced pressure. The title product was
afforded as a pale yellow solid.
[1832] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.57-7.50 (m, 3H),
7.50-7.43 (m, 2H), 4.01 (t, 2H), 3.93 (s, 3H), 3.36 (s, 3H), 2.72
(t, 2H), 2.22 (m, 2H).
[1833] LC-MS Rt 0.91 min [M+H].sup.+ 324 (Method: 2minLowpH)
Step 4:
1,3-Dimethyl-2,4-dioxo-5-phenyl-1,2,3,4,7,8-hexahydropyrimido[4,5--
a]indolizin-10-yl trifluoromethanesulfonate
[1834]
1,3-dimethyl-5-phenyl-8,9-dihydropyrimido[4,5-a]indolizine-2,4,10(1-
H,3H,7H)-trione (step 3) (920 mg, 2.85 mmol) and 2,6-lutidine
(0.497 ml, 4.27 mmol) were dissolved in DCM (25 ml) at 0.degree. C.
Trifluoromethanesulfonic anhydride (0.577 ml, 3.41 mmol) was added
slowly and the mixture stirred in an ice bath for 1.5 hours. The
reaction mixture was diluted with water and extracted with DCM. The
organic phase was passed through a hydrophobic frit and evaporated
under reduced pressure. Purification by chromatography on silica
eluting with 40% EtOAc/hexane yielded the title compound as a
yellow solid.
[1835] 1H NMR (400 MHz, CDCl3) .delta. 7.54-7.46 (m, 5H), 6.00 (t,
1H), 3.92 (t, 2H), 3.61 (s, 3H), 3.37 (s, 3H), 2.57 (dt, 2H);
[1836] LC-MS Rt 1.14 min [M+H].sup.+ 456 (Method: 2minLowpH).
Intermediate L
5-(3-Fluorophenyl)-10-hydroxy-1,3-dimethyl-7,8,9,10-tetrahydropyrimido[4,5-
-a]indolizine-2,4(1H,3H)-dione
##STR00183##
[1837] Step 1:
5-(3-Fluorophenyl)-1,3-dimethyl-8,9-dihydropyrimido[4,5-a]indolizine-2,4,-
10(1H,3H,7H)-trione
[1838] The title compound was prepared analogously to
3-(1,3-dimethyl-2,4,10-trioxo-1,2,3,4,7,8,9,10-octahydropyrimido[4,5-a]in-
dolizin-5-yl)benzonitrile (Example 9 steps 1-7) by replacing
3-bromobenzonitrile (step 3) with 1-bromo-3-fluorobenzene
(commercially available).
[1839] LC-MS: Rt 1.02 mins; MS m/z 342.5 [M+H]+; Method
2minLowpHv01
Step 2:
5-(3-Fluorophenyl)-10-hydroxy-1,3-dimethyl-7,8,9,10-tetrahydropyri-
mido[4,5-a]indolizine-2,4(1H,3H)-dione
[1840] To a stirred solution of
5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydropyrimido[4,5-a]indolizine-2,4,-
10(1H,3H,7H)-trione (step 1)(150 mg, 0.439 mmol) in MeOH (2197
.mu.L) at 0.degree. C. was added sodium borohydride (49.9 mg, 1.318
mmol) portionwise. The resulting yellow solution was warmed to room
temperature and stirred for 15 minutes. The reaction was
concentrated in vacuo. The residue was diluted with water (5 mL)
and DCM (10 mL). The aqueous phase was separated and extracted with
DCM (3.times.10 mL). The combined organic fractions were dried
(Na.sub.2SO.sub.4) and concentrated under reduced pressure to
afford the title compound as a pale yellow solid;
[1841] 1H NMR (400 MHz, DMSO-d6) .delta. 7.50 (1H, q), 7.33-7.24
(3H, m), 5.34-5.25 (1H, m), 5.22-5.15 (1H, m), 3.93-3.74 (2H, m),
3.70 (3H, s), 3.17 (3H, s), 2.19-2.04 (1H, m), 2.03-1.82 (2H, m),
1.82-1.74 (1H, m).
[1842] LCMS: Rt 0.94 mins; MS m/z 344.5 [M+H]+; Method
2minlowpHv01.
Intermediate La
5-(3-Fluorophenyl)-10-hydroxy-1,3,8-trimethyl-7,8,9,10-tetrahydropyrimido[-
4,5-a]indolizine-2,4(1H,3H)-dione
##STR00184##
[1843] Step 1:
5-(3-Fluorophenyl)-1,3,8-trimethyl-8,9-dihydropyrimido[4,5-a]indolizine-2-
,4,10(1H,3H,7H)-trione
[1844] The title compound was prepared analogously to
3-(1,3-dimethyl-2,4,10-trioxo-1,2,3,4,7,8,9,10-octahydropyrimido[4,5-a]in-
dolizin-5-yl)benzonitrile (Example 9 steps 1-7) by replacing
3-bromobenzonitrile (step 3) with 1-bromo-3-fluorobenzene
(commercially available) and methyl 4-bromobutanoate (Step 5) with
methyl 4-bromo-3-methylbutanoate (Intermediate T).
[1845] LC-MS Rt 1.05 mins; [M+H].sup.+ 356.5 (Method
2minLowpHv01)
Step 2:
5-(3-Fluorophenyl)-10-hydroxy-1,3,8-trimethyl-7,8,9,10-tetrahydrop-
yrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1846] The title compound was prepared analogously to Intermediate
L using by replacing
5-(3-fluorophenyl)-1,3-dimethyl-8,9-dihydropyrimido[4,5-a]indolizine-2,4,-
10(1H,3H,7H)-trione in (step 2) with
5-(3-fluorophenyl)-1,3,8-trimethyl-8,9-dihydropyrimido[4,5-a]indolizine-2-
,4,10(1H,3H,7H)-trione.
[1847] LC-MS Rt 1.01 mins [M+H].sup.+ 358.6 (Method
2minLowpHv01)
Intermediate Lb
5-(3-Fluorophenyl)-10-hydroxy-1,3,9-trimethyl-7,8,9,10-tetrahydropyrimido[-
4,5-a]indolizine-2,4(1H,3H)-dione
##STR00185##
[1849] The title compound was prepared analogously to
5-(3-fluorophenyl)-10-hydroxy-1,3,8-trimethyl-7,8,9,10-tetrahydropyrimido-
[4,5-a]indolizine-2,4(1H,3H)-dione (Intermediate La) by replacing
Methyl 4-bromo-3-methylbutanoate (Intermediate T) with methyl
4-bromo-2-methylbutanoate (Intermediate S).
[1850] LC-MS Rt 1.01 mins [M+H].sup.+ 358.6 (Method
2minLowpHv01)
Intermediate M
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-m-tolyl-7,8-dihydropyrimido[4,5-a-
]indolizine-2,4(1H,3H)-dione
##STR00186##
[1851] Step 1:
Ethyl-4-(1,3-dimethyl-2,4-dioxo-5-m-tolyl-3,4-dihydro-1H-pyrrolo[3,4-d]py-
rimidin-6(2H)-yl)butanoate
[1852] The title compound was prepared analogously to Intermediate
I, step 1 by replacing
5-benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
(Intermediate C) with
6-bromomethyl-1,3-dimethyl-5-(3-methyl-benzoyl)-1H-pyrimidine-2,4-dione
(Intermediate F).
[1853] .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 7.37 (1H, t),
7.27-7.20 (3H, mult), 6.40 (1H, s), 4.08 (2H, q), 3.96 (2H, t),
3.42 (3H, s), 3.35 (3H, s), 2.42 (3H, s), 2.19 (2H, t), 1.99 (2H,
t), 1.22 (3H, t);
[1854] LC-MS Rt=1.16 mins; [M+H].sup.+ 384.3 (Method
2minLowpHv01).
Step 2:
4-(1,3-Dimethyl-2,4-dioxo-5-m-tolyl-3,4-dihydro-1H-pyrrolo[3,4-d]p-
yrimidin-6(2H)-yl)butanoic acid
[1855] Lithium hydroxide (2.64 g, 63.0 mmol) was added to a
solution of ethyl
4-(1,3-dimethyl-2,4-dioxo-5-m-tolyl-3,4-dihydro-1H-pyrrolo[3,4-d]py-
rimidin-6(2H)-yl)butanoate (step 1) (4.83 g, 12.6 mmol) in THF (48
mL)/water (12 mL). The mixture was stirred at room temperature for
24 hours. The organic solvent was largely removed under vacuum, the
residue was acidified to pH 1, using 1M HCl. The precipitate was
extracted into chloroform (1.times.100 mL, 4.times.50 mL). The
combined organic extracts were dried with magnesium sulfate and the
solvent was removed under vacuum. The resulting solid was dried
under vacuum at 50.degree. C. to give the title compound.
[1856] .sup.1H NMR (400 MHz, CDCl.sub.3) d 7.37 (1H, t), 7.27-7.22
(3H, mult), 6.40 (1H, s), 3.98 (2H, t), 3.42 (3H, s), 3.35 (3H, s),
2.42 (3H, s), 2.25 (2H, t), 1.99 (2H, mult);
[1857] LC-MS Rt=0.96 mins; [M+H].sup.+ 356.3 (Method
2minLowpHv01).
Step 3:
1,3-Dimethyl-5-m-tolyl-8,9-dihydropyrimido[4,5-a]indolizine-2,4,10-
(1H,3H,7H)-trione
[1858] A solution of T3P.RTM. (1-propylphosphonic acid cyclic
anhydride 50% solution in DMF) (1.0M, 329 .mu.L, 0.56 mmol) was
added to solid
4-(1,3-dimethyl-2,4-dioxo-5-m-tolyl-3,4-dihydro-1H-pyrrolo[3,4-d]pyrimidi-
n-6(2H)-yl)butanoic acid (step 2) (200 mg, 0.56 mmol), giving a
paste. DMF (1 mL) was added and a solution formed. The solution was
stirred at room temperature for 1 hour and at 100.degree. C. for 1
hour. The reaction mixture was diluted with ethyl acetate (10 mL)
and washed with sat. sodium bicarbonate (1.times.5 mL) and brine
(3.times.5 mL). The organic phase was dried with magnesium sulfate
and the solvent was removed under vacuum to give the title
compound.
[1859] .sup.1H NMR (400 MHz, CDCl.sub.3) d 7.42 (1H, t), 7.34 (1H,
d), 7.27-7.23 (2H, mult), 4.00 (2H, mult), 3.93 (3H, s), 3.36 (3H,
s), 2.71 (2H, t), 2.45 (3H, s), 2.21 (2H, mult);
[1860] LC-MS Rt=1.05 mins; [M+H].sup.+ 338.4 (Method
2minLowpHv01).
Step 4:
1,3-Dimethyl-2,4-dioxo-5-m-tolyl-1,2,3,4,7,8-hexahydropyrimido[4,5-
-a]indolizin-10-yl trifluoromethanesulfonate
[1861] Trifluoromethanesulfonic anhydride (253 .mu.L, 1.50 mmol)
was added to an ice-cooled solution of
1,3-dimethyl-5-m-tolyl-8,9-dihydropyrimido[4,5-a]indolizine-2,4,10(1H,3H,-
7H)-trione (step 3) (388 mg, 1.15 mmol) and 2,6-lutidine (201
.mu.L, 1.73 mmol) in DCM (7 mL). The solution was allowed to reach
room temperature slowly and was stirred for 16 hours. A further
portion of trifluoromethanesulfonic anhydride (39 .mu.L, 0.23 mmol)
was added and the solution was stirred for a further 30 minutes.
The reaction mixture was diluted with dichloromethane (15 mL) and
washed with water (2.times.8 mL) and sat brine (1.times.8 mL). The
organic phase was dried with magnesium sulfate and the solvent was
removed under vacuum, keeping the rotary evaporator water bath at
20.degree. C. to give the title compound.
[1862] LC-MS Rt=1.31 min; [M+H].sup.+ 470.5 (Method
2minLowpHv01).
Step 5:
1,3-Dimethyl-10-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-m--
tolyl-7,8-dihydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione
[1863] A mixture of
1,3-dimethyl-2,4-dioxo-5-m-tolyl-1,2,3,4,7,8-hexahydropyrimido[4,5-a]indo-
lizin-10-yl trifluoromethanesulfonate (unpurified material from
step 4, assumed 1.15 mmol), bis(pinaolato)diboron (321 mg, 1.27
mmol), bis(triphenylphosphine)palladium(II) chloride (24 mg, 0.035
mmol), triphenylphosphine (18 mg, 0.07 mmol) and potassium
phenoxide (J. Am. Chem. Soc., 1959, Vol. 81, pp 2705-2715, 228 mg,
1.73 mmol) in toluene (8 mL) was stirred at 60.degree. C., under
nitrogen for 4 hours. The reaction mixture was diluted with water
(20 mL) and was extracted with ethyl acetate (3.times.20 mL). The
combined organic extracts were washed with brine (1.times.20 mL),
dried with magnesium sulfate and the solvent was removed under
vacuum. Purification by chromatography on silica, eluting with
20-50% EtOAc/hexane gave the product as an oil. Addition of
iso-hexane and subsequent removal under vacuum gave the title
compound as an off-white solid.
[1864] 1H NMR (400 MHz, CDCl.sub.3) .delta. 7.35 (1H, t), 7.28-7.22
(3H, mult), 6.75 (1H, t), 3.79 (2H, t), 3.58 (3H, s), 3.35 (3H, s),
2.41 (3H, s), 2.38 (2H, mult), 1.35 (12H, s);
[1865] LC-MS Rt=1.36 min [M+H].sup.+ 448.6 (Method
2minLowpHv01).
Step 6:
1,3-Dimethyl-10-(4-methylthiazol-2-yl)-5-m-tolyl-7,8-dihydropyrimi-
do[4,5-a]indolizine-2,4(1H,3H)-dione
[1866] A mixture of
1,3-dimethyl-10-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-m-tolyl-7-
,8-dihydropyrimido[4,5-a]indolizine-2,4(1H,3H)-dione (step 5) (295
mg, 0.66 mmol), 2-iodo-4-methylthiazole (Intermediate 0) (135 mg,
0.60 mmol), Pd-118 (19.6 mg, 0.03 mmol) and barium hydroxide (206
mg, 1.20 mmol) in acetonitrile/water (1:1, 3 mL) was stirred at
80.degree. C. for 30 minutes. The reaction mixture was diluted with
1M HCl (5 mL) and extracted with DCM (3.times.5 mL). The combined
organic layers were filtered through a hydrophobic frit and the
solvent was removed under vacuum. The residue was purified by
chromatography on silica, eluting with 1-2% MeOH/DCM to give the
title compound.
[1867] LC-MS Rt=1.24 mins; [M+H].sup.+ 419.1 (Method
2minLowpHv01).
Intermediate Na
1,3-Dimethyl-2,4-dioxo-5-phenyl-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimi-
dine-7-carbaldehyde
##STR00187##
[1868] Step 1:
1,3-Dimethyl-5-phenyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1869]
5-Benzoyl-6-(bromomethyl)-1,3-dimethylpyrimidine-2,4(1H,3H)-dione
13.4a (Intermediate C) (2.33 g, 6.91 mmol), hexamethyldisilazane
(2.90 mL, 13.82 mmol) and triethylamine (2.89 mL, 20.73 mmol) were
combined in dioxane (20 mL) and the mixture heated at reflux for 22
hours. Further portions of hexamethyldisilazane (2.90 mL, 13.82
mmol) and triethylamine (2.89 mL, 20.73 mmol) were added to allow
the reaction to run to completion. The reaction mixture was cooled
to room temperature, diluted with EtOAc (150 mL) and washed with 1M
HCl (aq) and brine. The organic phases was dried over sodium
sulphate and evaporated under vacuum. The residue was triturated
with DCM/diethyl ether/hexane to afford the title compound.
[1870] LC-MS Rt 0.98 mins; MS 256.1 m/z [M+H] Method
2minLowpHv03
Step 2:
1,3-Dimethyl-2,4-dioxo-5-phenyl-2,3,4,6-tetrahydro-1H-pyrrolo[3,4--
d]pyrimidine-7-carbaldehyde
[1871] Phosphorous oxychloride (1.081 mL, 11.60 mmol) was added
slowly to a solution of
1,3-dimethyl-5-phenyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
(2.96 g, 11.60 mmol) in DMF (40 mL) at 0.degree. C. The mixture was
stirred at 0.degree. C. for 15 mins. The reaction was quenched with
2M HCl(aq), diluted with EtOAc, and the mixture stirred overnight.
The mixture was then extracted with EtOAc (3.times.) and the
combined organic phases washed with brine, dried over sodium
sulphate and evaporated under vacuum. The residue was triturated
with diethyl ether/hexane to afford the title compound.
[1872] LC-MS Rt 0.97 mins; MS 284.2 m/z [M+H] Method
2minLowpHv03
Intermediate Nb
3-(7-Formyl-1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyr-
imidin-5-yl)benzonitrile
##STR00188##
[1874] The title compound was prepared in a similar manner to
1,3-dimethyl-2,4-dioxo-5-phenyl-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrim-
idine-7-carbaldehyde (Intermediate Na), starting from
3-(1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-
-yl)benzonitrile (Example 9.0 Step 4) and omitting step 1.
[1875] LC-MS Rt 0.93 mins [M+H].sup.+ 309.2 (Method
2minLowpHv03)
Intermediate Nc
5-(3-Fluorophenyl)-1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,-
4-d]pyrimidine-7-carbaldehyde
##STR00189##
[1877] The title compound was prepared in a similar manner to
1,3-dimethyl-2,4-dioxo-5-phenyl-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrim-
idine-7-carbaldehyde (Intermediate Na), starting from
5-(3-fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-di-
one (prepared in a similar manner to
3-(1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-
-yl)benzonitrile (Example 9.0 Step 4) using the appropriate halo
compound in step 3 and omitting Step 1.
Intermediate Nd
1,3-Dimethyl-5-(4-methylthiazol-2-yl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6-
H)-dione
##STR00190##
[1879] The title compound was prepared analogously to
3-(1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-
-yl)benzonitrile (Example 9.0 Step 4) by replacing
3-bromobenzonitrile by 2-iodo-4-methylthiazole (Intermediate O) in
step 3;
[1880] LC-MS: Rt 0.95 mins [M+H].sup.+ 277.4 (Method
2minLowpHv03).
Intermediate Ne
5-(3-Fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dio-
ne
##STR00191##
[1882] The title compound was prepared analogously to
3-(1,3-Dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-
-yl)benzonitrile (Example 9.0 Step 4) by replacing
3-bromobenzonitrile by 3-fluoro-1-bromobenzene in step 3;
[1883] LC-MS Rt 1.06 mins [M+H].sup.+ 274.1 (Method
2minLowpHv03)
Intermediate Nf
5-(3-Fluorophenyl)-1,3-dimethyl-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dio-
ne
##STR00192##
[1885] The title compound was prepared by an analogous method to
3-(1,3-dimethyl-2,4-dioxo-2,3,4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-
-yl)benzonitrile (Ex. 9 step 4) by replacing 3-bromobenzonitrile
(step 3) with 1-bromo-3-fluorobenzene (commercially available).
LC-MS Rt 1.04 mins [M+H].sup.+ 274.5 (Method 2minLowpHv03).
Intermediate O
2-Iodo-4-methylthiazole
##STR00193##
[1887] Methyllithium solution (1.6 M in Et.sub.2O, 189 ml, 303
mmol) was added dropwise to 4-methylthiazole (22.94 ml, 252 mmol)
in diethyl ether (505 ml) at -78.degree. C. The mixture was stirred
for 30 mins at -78.degree. C., then iodine (83 g, 328 mmol) was
added and the suspension slowly warmed to 0.degree. C. and stirred
for 45 mins. The reaction was quenched by the addition of water
(150 ml), diluted with diethyl ether (100 ml) and the layers
separated. The aqueous phase was extracted with diethyl ether
(2.times.200 ml) and the combined organic phases were washed with
saturated aqueous sodium thiosulfate solution (150 ml), 2M
Na.sub.2CO.sub.3(aq) (100 ml), water (100 ml) and brine (100 ml),
then dried over magnesium sulfate and evaporated under vacuum.
Purification by chromatography on silica, eluting with 0-5%
TBME/iso-hexane afforded the title compound.
[1888] 1H NMR (400 MHz, CDCl3) .delta. 6.87 (1H, s), 2.48 (3H,
s)
[1889] LC-MS Rt 1.00 mins [M+H].sup.+ 2.26 (Method
2minLowpHv03)
Intermediate P
4-Bromo-N-methoxy-N-methylbutanamide
##STR00194##
[1891] 4-Bromobutanoyl chloride (5.62 ml, 48.5 mmol) in DCM (64 ml)
was added slowly to a solution of DIPEA (17.80 ml, 102 mmol) and
N,O-dimethylhydroxylamine hydrochloride (4.97 g, 51.0 mmol) in DCM
(32 ml) at 0.degree. C. The mixture was stirred at room temperature
for 16 hours then diluted with DCM (50 ml). The mixture was washed
with 1M HCl(aq) (2.times.50 ml) and saturated NaHCO.sub.3(aq)
(2.times.50 ml). The organic phase was passed through a hydrophobic
frit and evaporated under vacuum. Purification by chromatography on
silica, eluting with 30% EtOAc/hexane afforded the title
compound.
[1892] 1H NMR (400 MHz, CDCl3) .delta. 3.69 (3H, s), 3.62 (2H, t),
3.49 (3H, s), 3.17 (3H, s), 2.61 (2H, t), 2.17 (2H, quint), 2.09
(2H, quint).
Intermediate Q
4-Chlorothiazole-2-carbaldehyde
##STR00195##
[1893] Step 1: 2,5-dibromo-4-chlorothiazole
[1894] 2,4-Dichlorothiazole (10 g, 64.9 mmol) was dissolved in
acetic acid (50.0 ml) and heated to 60.degree. C. Bromine (15.05
ml, 292 mmol) was added dropwise and the reaction mixture stirred
at 90.degree. C. for 5.5 hours. The mixture was cooled to room
temperature and made basic by slow addition of solid sodium
carbonate, then diluted with water (100 mL) and extracted with
Et.sub.2O (3.times.150 ml). The combined organic extracts were
washed with sat. aq. sodium thiosulfate (50 ml) dried over sodium
sulfate evaporated under reduced pressure to afford the title
compound as a pale yellow oil.
[1895] LC-MS Rt 1.43 min [M+H].sup.+ 275.9/277.9/279.9/281.9
(Method 2minLowpHv03)
Step 2: 2-Bromo-4-chlorothiazole
[1896] nBuLi (2.5M in hexanes, 7.21 mL, 18.03 mmol) was added
dropwise over 20mins to 2,5-dibromo-4-chlorothiazole (5 g, 18.03
mmol) in THF (100 mL) at -90.degree. C., and the mixture stirred at
-90.degree. C. for 30 mins. Water (0.341 mL, 18.93 mmol) in THF (2
mL) was added and the mixture stirred whilst slowly warming to room
temperature. The reaction was quenched with 0.1M HCl(aq) (100 mL)
and extracted with Et.sub.2O (150 ml). The organic phase was washed
with saturated NaHCO.sub.3(aq) (100 mL) and water (100 mL), dried
over magnesium sulphate and evaporated under vacuo to afford the
product as a pale yellow oil which slowly crystallised as pale
yellow needles.
[1897] 1H NMR: (400 MHz, CDCl.sub.3) .delta. 7.09 (1H, s).
[1898] LC-MS: Rt 1.12mins [M+H].sup.+ 199.9 (Method
2minLowpHv03)
Step 3: 4-Chlorothiazole-2-carbaldehyde
[1899] n-Butyllithium (1.732 ml, 2.77 mmol) was added dropwise over
20 mins to a solution of 2-bromo-4-chlorothizaole (500 mg, 2.52
mmol) in diethyl ether (25.2 ml) at -78.degree.. The mixture was
stirred at -78.degree. C. for 20, DMF (0.215 ml, 2.77 mmol) was
added and the mixture warmed to -35.degree. C. over 40 mins. The
reaction was quenched with 6M HCl and the phases separated. The
aqueous phase was extracted with diethyl ether (20 ml). The aqueous
layer was brought to pH 10 by addition of solid potassium carbonate
at 5.degree. C. and extracted with diethyl ether (3.times.35 ml).
The combined organic extracts were dried over magnesium sulfate and
evaporated under vacuum. Purification by chromatography on silica,
eluting with hexane, afforded the title compound.
[1900] 1H NMR (400 MHz, CDCl.sub.3) .delta. 9.98 (1H, s), 7.59 (1H,
s).
Intermediate R
5-(4-(((4-Methoxybenzyl)oxy)methyl)thiazol-2-yl)-1,3-dimethyl-1H-pyrrolo[3-
,4-d]pyrimidine-2,4(3H,6H)-dione
##STR00196##
[1901] Step 1: Ethyl
2-(1,3-dimethyl-2,4-dioxo-6-((2-(trimethylsilyl)ethoxy)methyl)-2,3,4,6-te-
trahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)thiazole-4-carboxylate
[1902] The title compound was prepared by a method analogous to
3-(1,3-dimethyl-2,4-dioxo-6-((2-(trimethylsilyl)ethoxy)methyl)-2,3,4,6-te-
trahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)benzonitrile (Ex. 9 step
3) using
1,3-dimethyl-2,4-dioxo-6-((2-(trimethylsilyl)ethoxy)methyl)-2,3,4,6-tetra-
hydro-1H-pyrrolo[3,4-d]pyrimidin-5-ylboronic acid (Example 9 Step
2) and the appropriate commercially available halo compound.
[1903] 1H NMR (400 MHz, DMSO-d6) .delta. 8.64 (1H, s), 7.33 (1H,
s), 5.94 (2H, s), 4.33 (2H, q), 3.44 (2H, t), 3.34 (3H, s), 3.24
(3H, s), 1.32 (3H, t), 0.75 (2H, t), -0.15 (9H, s)
[1904] LC-MS Rt 1.55 mins [M+H].sup.+ 465.3 (Method
2minLowpHv03)
Step 2:
5-(4-(Hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-6-((2-(trimethylsil-
yl)ethoxy)methyl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1905] Sodium borohydride (0.856 g, 22.62 mmol) was added to a
solution of ethyl
2-(1,3-dimethyl-2,4-dioxo-6-((2-(trimethylsilyl)ethoxy)methyl)-2,3,-
4,6-tetrahydro-1H-pyrrolo[3,4-d]pyrimidin-5-yl)thiazole-4-carboxylate
(step 1) (5.254 g, 11.31 mmol) and lithium chloride (0.959 g, 22.62
mmol) in ethanol (145 ml) and THF (290 ml) at 0.degree. C. The
mixture was stirred at room temperature for 16 hours. Further
portions of lithium chloride (0.959 g, 22.62 mmol) and sodium
borohydride (0.856 g, 22.62 mmol) were added and the mixture
stirred for a further 4 hours. The reaction was quenched with
saturated NaHCO.sub.3 (aq) (250 ml), diluted with water (100 ml)
and extracted with chloroform (3.times.350 ml). The combined
organic extracts were washed with water (200 ml) and brine
(2.times.200 ml), dried over sodium sulfate and evaporated under
vacuum to afford the title compound.
[1906] 1H NMR (400 MHz, DMSO-d6) .delta. 7.60 (1H, s), 7.23 (1H,
s), 5.80 (2H, s), 5.38 (1H, t), 4.63 (2H, dd), 3.37 (2H, m), 3.33
(3H, s), 3.22 (3H, s), 0.73 (2H, m), -0.12 (9H, s).
[1907] LC-MS Rt 1.28 mins [M+H].sup.+ 423.2 (Method
2minLowpHv03)
Step 3:
5-(4-(((4-Methoxybenzyl)oxy)methyl)thiazol-2-yl)-1,3-dimethyl-6-((-
2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-di-
one and
5-(4-(((4-methoxybenzyl)oxy)methyl)thiazol-2-yl)-1,3-dimethyl-1H-p-
yrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1908] Sodium hydride (60% wt, 0.637 g, 15.93 mmol) was added
portionwise to a solution of
5-(4-(hydroxymethyl)thiazol-2-yl)-1,3-dimethyl-6-((2-(trimethylsilyl)etho-
xy)methyl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione (step 2)
(4.43 g, 7.97 mmol) NMP (Volume: 44 ml) at 0.degree. C. and stirred
for 1 hour. para-Methoxybenzyl chloride (1.404 ml, 10.36 mmol) was
added and the mixture stirred at room temperature for 5 hours.
Further portions of para-methoxybenzyl chloride (220 .mu.l) and
sodium hydride (60% wt, 64 mg) were added and the mixture stirred
for a further 16 hours. The reaction was quenched at 0.degree. C.
with saturated NaHCO.sub.3(aq) (50 ml), diluted with water (200 ml)
and EtOAc (100 ml) and the phases separated. The aqueous phase was
extracted with EtOAc (3.times.100 ml). The combined organic
extracts were washed with water (2.times.100 ml) and brine (100
ml), dried over sodium sulfate and evaporated under vacuum.
Purification by chromatography on silica, eluting with 40-60%
EtOAc/hexane afforded the title compound and
5-(4-(((4-methoxybenzyl)oxy)methyl)thiazol-2-yl)-1,3-dimethyl-1H-pyrrolo[-
3,4-d]pyrimidine-2,4(3H,6H)-dione as a mixture which was used
without further purification.
[1909] LC-MS Rt 1.63 mins [M+H].sup.+ 543.6 (Method
2minLowpHv03)
Step 5:
5-(4-(((4-Methoxybenzyl)oxy)methyl)thiazol-2-yl)-1,3-dimethyl-1H-p-
yrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
[1910] tetra-n-Butylammonium fluoride solution (1.0 M in THF, 48.3
ml, 48.3 mmol) was added to a solution of
5-(4-(((4-methoxybenzyl)oxy)methyl)thiazol-2-yl)-1,3-dimethyl-6-((2-(trim-
ethylsilyl)ethoxy)methyl)-1H-pyrrolo[3,4-d]pyrimidine-2,4(3H,6H)-dione
and
5-(4-(((4-methoxybenzyl)oxy)methyl)thiazol-2-yl)-1,3-dimethyl-1H-pyrrolo[-
3,4-d]pyrimidine-2,4(3H,6H)-dione (5.24 g) in THF (Volume: 13.91
ml) at 60.degree. C. The mixture was stirred at 60.degree. C. for 6
hours. The mixture was then cooled to room temperature and
evaporated under vacuum. The residue partitioned between EtOAc (100
ml) and water (150 ml) and the phases were separated. The organic
phase was washed with water (3.times.200 ml) and both phases were
stood for 60 hours. Both phases were filtered and the solids dried
under vacuum to afford the title compound.
[1911] 1H NMR (400 MHz, DMSO-d6) .delta. 12.79 (1H, s), 7.76 (1H,
s), 7.45 (2H, d), 7.12-7.05 (3H, m), 4.77 (2H, s), 4.68 (2H, s),
3.90 (3H, s), 3.48 (3H, s), 3.40 (3H, s)
[1912] LC-MS Rt 1.24 mins [M+H].sup.+ 413.5 (Method
2minLowpHv03)
Intermediate S
Methyl 4-bromo-2-methylbutanoate
##STR00197##
[1914] Boron tribromide solution (44.4 mL, 1.0M in DCM, 44.4 mmol)
was added dropwise to alpha-methyl-gammabutyrolactone (commercial)
(4.23 g, 42.3 mmol) in DCM (47 mL) at 0.degree. C. The mixture was
warmed to room temperature and stirred for 19 hours. The reaction
was quenched with methanol (10.5 mL) at 0.degree. C. and stirred at
room temperature for 30 minutes, then diluted with water (100 mL)
and extracted with DCM (1.times.50 mL). The organic extracts were
washed with saturated NaHCO.sub.3(aq) (2.times.50 mL) dried over
magnesium sulfate and evaporated under vacuum. Purification by
chromatography on silica afforded the title compound.
[1915] 1H NMR (400 MHz, CDCl.sub.3) .delta. 3.72 (3H, s), 3.44 (2H,
br t), 2.74 (1H, mult), 2.29 (1H, mult), 1.94 (1H, mult), 1.22 (3H,
mult)
Intermediate Sa
(S)-(2,2-Dimethyl-1,3-dioxolan-4-yl)methyl
trifluoromethanesulfonate
##STR00198##
[1917] Trifluoromethanesulfonic anyhdride (1.398 ml, 8.32 mmol) was
added dropwise to a solution of commercially available R-solketal
(1 g, 7.57 mmol) and 2.6-lutidine (1.139 ml, 9.84 mmol) in DCM
(25.2 mL) at 0.degree. C. The mixture was warmed to room
temperature and stirred for 1 hour. The mixture was then evaporated
under vacuum and the residue partitioned between water (10 mL) and
DCM (20 mL). The phases were separated and the aqueous phase was
extracted with DCM (3.times.10 mL). The combined organic extracts
were dried over magnesium sulfate to afford the title compound as a
crude material which was used without further purification.
Intermediate Sb
(2,2-Dimethyl-1,3-dioxolan-4-yl)methyl
trifluoromethanesulfonate
##STR00199##
[1919] The title compound was prepared by an analogous method to
Intermediate Ea from commercially available racemic solketal.
Intermediate T
Methyl 4-bromo-3-methylbutanoate
##STR00200##
[1920] Step 1: 4-Methyldihydrofuran-2(3H)-one
[1921] A mixture of 4-methylfuran-2(5H)-one (800 mg, 8.2 mmol) and
10% palladium on carbon (80 mg) in EtOAc (17 mL) was stirred under
an atmosphere of hydrogen for 18 hours. The mixture was filtered
and the residue rinsed with EtOAc. The filtrates were evaporated
under vacuum to afford the title compound.
[1922] 1H NMR (400 MHz, CDCl.sub.3) .delta. 4.40 (1H, mult), 3.86
(1H, mult), 2.69-2.59 (2H, mult), 2.13 (1H, mult), 1.15 (3H,
d).
Step 2: Methyl 4-bromo-3-methylbutanoate
[1923] Boron tribromide solution (9.1 mL, 1.0M in DCM, 9.1 mmol)
was added dropwise to a solution of 4-methyldihydrofuran-2(3H)-one
(870 mg, 8.7 mmol) in DCM (9 mL) at 0.degree. C. The mixture was
stirred at room temperature for 20 hours. The reaction mixture was
re-cooled to 0.degree. C. and methanol (2 mL) was added dropwise.
The mixture was warmed to room temperature and stirred for a
further 30 minutes. The reaction was quenched with water (20 mL)
and extracted with DCM (3.times.10 mL). The combined organic
extracts were washed with saturated NaHCO.sub.3(aq) (1.times.10
mL), dried over magnesium sulfate and evaporated under vacuum.
Purification by chromatography on silica, eluting with 20-50%
DCM/hexane, afforded the title compound.
[1924] 1H NMR (400 MHz, CDCl.sub.3) .delta. 3.70 (3H, s), 3.44 (2H,
mult), 2.54 (1H, mult), 2.38-2.23 (2H, mult), 1.08 (3H, d).
* * * * *