U.S. patent application number 11/751451 was filed with the patent office on 2007-09-13 for 2-nh-heteroarylimidazoles with antibacterial activity.
This patent application is currently assigned to Replidyne, Inc.. Invention is credited to John Berge, Andrew Forrest, Dieter Hamprecht, Richard Jarvest.
Application Number | 20070213362 11/751451 |
Document ID | / |
Family ID | 9949239 |
Filed Date | 2007-09-13 |
United States Patent
Application |
20070213362 |
Kind Code |
A1 |
Berge; John ; et
al. |
September 13, 2007 |
2-NH-HETEROARYLIMIDAZOLES WITH ANTIBACTERIAL ACTIVITY
Abstract
A method of inhibiting MetRS activity comprises administering to
a patient in need thereof a MetRS inhibiting effective amount of a
compound of the formula (I). A method of treating a resistant or
multiply-resistant E. faecalis infection, a resistant or
multiply-resistant S. aureus infection, and/or a resistant or
multiply-resistant S. epidermidis infection comprises administering
to a patient in need thereof an antibacterially effective amount of
a compound of the formula (I).
Inventors: |
Berge; John; (Essex, GB)
; Forrest; Andrew; (Essex, GB) ; Hamprecht;
Dieter; (Verona, IT) ; Jarvest; Richard;
(Hertfordshire, GB) |
Correspondence
Address: |
SWANSON & BRATSCHUN, L.L.C.
8210 SOUTHPARK TERRACE
LITTLETON
CO
80120
US
|
Assignee: |
Replidyne, Inc.
Louisville
CO
|
Family ID: |
9949239 |
Appl. No.: |
11/751451 |
Filed: |
May 21, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
11223327 |
Sep 9, 2005 |
7220757 |
|
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11751451 |
May 21, 2007 |
|
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10729416 |
Dec 5, 2003 |
6943175 |
|
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11223327 |
Sep 9, 2005 |
|
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Current U.S.
Class: |
514/303 ;
514/393 |
Current CPC
Class: |
C07D 487/04 20130101;
A61K 31/44 20130101; A61K 31/4164 20130101; C07D 495/04 20130101;
C07D 473/00 20130101; C07D 471/04 20130101; Y02A 50/473 20180101;
A61P 31/04 20180101; Y02A 50/30 20180101 |
Class at
Publication: |
514/303 ;
514/393 |
International
Class: |
A61K 31/4164 20060101
A61K031/4164; A61K 31/44 20060101 A61K031/44 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 6, 2002 |
GB |
0228545.0 |
Claims
1. A method of inhibiting MetRS activity comprising administering
to a patient in need thereof a MetRS inhibiting effective amount of
a compound of the formula (I): ##STR92## in which: R.sup.1 is an
optionally substituted aryl or an optionally substituted heteroaryl
ring; R.sup.2 is a 5 or 6-membered heteroaryl ring which is
optionally substituted with from 1 to 3 substituents selected from
halo, cyano, hydroxy, (C.sub.1-6)alkyl (optionally substituted by
halo, hydroxy, amino, mono to perfluoro(C.sub.1-3)alkyl, carboxy or
(C.sub.1-6)alkoxycarbonyl), (C.sub.3-7)cycloalkyl,
C.sub.(1-6)alkoxy, amino, mono- or di-(C.sub.1-6)alkylamino,
acylamino, carboxy, (C.sub.1-6)alkoxycarbonyl,
carboxy(C.sub.1-6)alkyloxy, (C.sub.1-6)alkylthio,
(C.sub.1-6)alkylsulphinyl, (C.sub.1-6)alkylsulphonyl, sulphamoyl,
mono- and di-(C.sub.1-6)alkylsulphamoyl, carbamoyl, mono- and
di-(C.sub.1-6)alkylcarbamoyl, and heterocyclyl; X is CH.sub.2 or
CHR.sup.3 in which R.sup.3 is C.sub.(1-6)alkyl or is linked to the
ortho position of an aryl or heteroaryl ring of R.sup.1 to form a 5
to 7 membered ring optionally including oxygen or nitrogen as a
ring atom; Y is C.sub.(1-3)alkylene or C.sub.(4-6)cycloalkylene;
including tautomeric forms of the imidazole ring; and salts thereof
and excluding 8-[2-(benzylamino)ethylamino]theophylline.
2. A method of inhibiting metRS activity according to claim 1,
wherein said compound is selected from the group consisting of
N-(3,5-dibromobenzyl)-N'-(1H-imidazo[4,5-b]pyridin-2-yl)-propane-1,3-diam-
ine;
N-(4,6-dichloro-1H-indol-2-ylmethyl)-N'-(H-imidazo[4,5-b]pyridin-2-y-
l)-propane-1,3-diamine;
N-(6,8-dibromo-1,2,3,4,-tetrahydroquinolin-4-yl)-N'-(1H-imidazo[4,5-b]pyr-
idin-2-yl)-propane-1,3-diamine dihydrochloride;
N-(4,5-dibromothien-2-ylmethyl)-N'-(H-imidazo[4,5-b]pyridin-2-yl)-propane-
-1,3-diamine;
N-(3,5-dibromobenzyl)-N'-(1H-imidazo[4,5-c]pyridin-2-yl)-propane-1,3-diam-
ine;
N-(4,6-dichloro-1H-indol-2-ylmethyl)-N'-(1H-imidazo[4,5-b]pyrazin-2--
yl)-propane-1,3-diamine dihydrochloride;
N-(4,6-dichloro-1H-indol-2-ylmethyl)-N'-(9H-purin-8-yl)-propane-1,3-diami-
ne dihydrochloride;
N-(4,5-dibromothien-2-ylmethyl)-N'-(9H-purin-8-yl)-propane-1,3-diamine
dihydrochloride;
N-(3-bromo-5-methoxy-1H-indol-7-ylmethyl)-N'-(1H-imidazo[4,5-b]pyridin-2--
yl)-propane-1,3-diamine;
N-(6-Ethyl-8-iodo-1,2,3,4-tetrahydroquinolin-4-yl)-N'-(1H-imidazo[4,5-b]p-
yridin-2-yl)-propane-1,3-diamine dihydrochloride;
N-(6,8-dibromo-1,2,3,4-tetrahydroquinolin-4-yl)-N'-(1H-imidazo[4,5-b]pyri-
din-2-yl)-propane-1,3-diamine dihydrochloride;
N-(3-chloro-5-methoxy-1H-indol-7-ylmethyl)-N'-(H-imidazo[4,5-b]pyridin-2--
yl)-propane-1,3-diamine;
N-(4,5-dibromo-3-methylthiophen-2-ylmethyl)-N'-(H-imidazo[4,5-c]pyridin-2-
-yl)propane-1,3-diamine;
N-(4,5-dibromo-3-methylthiophen-2-ylmethyl)-N'-(5H-imidazo[4,5-c]pyridazi-
n-6-yl)propane-1,3-diamine dihydrochloride;
N-(4,5-dibromo-3-methylthiophen-2-ylmethyl)-N'-(H-imidazo[4,5-b]pyridin-2-
-yl)propane-1,3-diamine;
N-(4-bromo-3-methyl-5-vinylthiophen-2-ylmethyl)-N'-(5H-imidazo[4,5-c]pyri-
dazin-6-yl)-propane-1,3-diamine;
N-(4-bromo-3-methyl-5-vinylthiophen-2-ylmethyl)-N'(1H-imidazo[4,5-b]pyrid-
in-2-yl)-propane-1,3-diamine;
N-(4-bromo-3-methyl-5-vinylthiophen-2-ylmethyl)-N'(1H-imidazo[4,5-c]pyrid-
in-2-yl)-propane-1,3-diamine;
N-(3-bromo-2-ethoxy-5-methylsulfanybenzyl)-N'-(5H-imidazo[4,5-c]pyridazin-
-6-yl)propane-1,3-diamine dihydrochloride;
N-(6-chloro-8-iodochroman-4-yl)-N'-(5H-imidazo[4,5-c]pyridazin-6-yl)propa-
ne-1,3-diamine;
N-(3-chloro-5-methoxy-1H-indol-7-ylmethyl)-N'-(5H-imidazo[4,5-c]pyridazin-
-6-yl)propane-1,3-diamine dihydrochloride;
N-(4,5-dibromo-3-methylthiophen-2-ylmethyl)-N'-(1H-thieno[3,4-d]imidazol--
2-yl)propane-1,3-diamine;
N-(4-bromo-3-methyl-5-vinylthiophen-2-ylmethyl)-N'-(1H-thieno[3,4-d]imida-
zol-2-yl)propane-1,3-diamine;
N-(3-chloro-5-methoxy-1H-indol-7-ylmethyl)-N'-(1H-thieno[3,4-d]imidazol-2-
-yl)propane-1,3-diamine;
N-(4-bromo-5-difluoromethyl-3-methylthiophen-2-ylmethyl)-N'-(1H-imidazo[4-
,5-b]pyridin-2-yl)propane-1,3-diamine dihydrochloride;
N-(4-bromo-5-difluoromethyl-3-methylthiophen-2-ylmethyl)-N'-(5H-imidazo[4-
,5-c]pyridazin-6-yl)propane-1,3-diamine dihydrochloride;
N-(4-bromo-5-difluoromethyl-3-methylthiophen-2-ylmethyl)-N'-(1H-thieno[3,-
4-d]imidazol-2-yl)propane-1,3-diamine dihydrochloride;
N-(4-bromo-3-methyl-5-trifluoromethylthiophen-2-ylmethyl)-N'-(5H-imidazo[-
4,5-c]pyridazin-6-yl)propane-1,3-diamine dihydrochloride;
N-(4-bromo-3-methyl-5-trifluoromethylthiophen-2-ylmethyl)-N'-(1H-thieno[3-
,4-d]imidazol-2-yl)propane-1,3-diamine dihydrochloride;
N-(4-bromo-3-methyl-5-trifluoromethylthiophen-2-ylmethyl)-N'-(1H-imidazo[-
4,5-b]pyridin-2-yl)propane-1,3-diamine dihydrochloride;
N-(4-bromo-3-methyl-5-trifluoromethylthiophen-2-ylmethyl)-N'-(1H-imidazo[-
4,5-c]pyridin-2-yl)propane-1,3-diamine dihydrochloride;
N-(4-bromo-5-(1-fluorovinyl)-3-methylthiophen-2-ylmethyl)-N'-(1H-imidazo[-
4,5-b]pyridin-2-yl)propane-1,3-diamine hydrochloride;
N-(4-bromo-5-(1-fluorovinyl)-3-methylthiophen-2-ylmethyl)-N'-(1H-imidazo[-
4,5-c]pyridin-2-yl)propane-1,3-diamine;
N-(4-bromo-5-(1-fluorovinyl)-3-methylthiophen-2-ylmethyl)-N'-(5H-imidazo[-
4,5-c]pyridazin-6-yl)propane-1,3-diamine;
N-(4-bromo-5-(1-fluorovinyl)-3-methylthiophen-2-ylmethyl)-N'-(1H-thieno[3-
,4-d]imidazol-2-yl)propane-1,3-diamine;
N-(4-bromo-3-methyl-5-ethynylthiophen-2-ylmethyl)-N'-(5H-imidazo[4,5-c]py-
ridazin-6-yl)propane-1,3-diamine;
N-(4-bromo-3-methyl-5-ethynylthiophen-2-ylmethyl)-N'-(1H-imidazo[4,5-b]py-
ridin-2-yl)propane-1,3-diamine;
N-(4-bromo-3-methyl-5-ethynylthiophen-2-ylmethyl)-N'-(1H-imidazo[4,5-c]py-
ridin-2-yl)propane-1,3-diamine;
N-(4-bromo-3-methyl-5-ethynylthiophen-2-ylmethyl)-N'-(1H-thieno[3,4-d]imi-
dazol-2-yl)propane-1,3-diamine;
N-(4-bromo-5-ethyl-3-(1-propynyl)thiophen-2-ylmethyl)-N'-(5H-imidazo[4,5--
c]pyridazin-6-yl)propane-1,3-diamine dihydrochloride;
N-(4-bromo-5-ethyl-3-(1-propynyl)thiophen-2-ylmethyl)-N'-(1H-imidazo[4,5--
c]pyridin-2-yl)propane-1,3-diamine;
N-(4-bromo-5-ethyl-3-(1-propynyl)thiophen-2-ylmethyl)-N'-(1H-imidazo[4,5--
b]pyridin-2-yl)propane-1,3-diamine; and
N-(4-bromo-5-ethyl-3-(1-propynyl)thiophen-2-ylmethyl)-N'-(1H-thieno[3,4-d-
]imidazol-2-yl)propane-1,3-diamine.
3. The method of claim 1, wherein the metRS is a metRS from a Gram
positive organism.
4. The method of claim 3, wherein the Gram positive organism is
selected from the group consisting of S. aureus, S. epidermidis, S.
pyogenes, S. pneumoniae, and Ent. faecalis.
5. The method of claim 1, wherein the metRS is a metRS from a Gram
negative organism.
6. The method of claim 5, wherein the Gram negative organism is
selected from the group consisting of H. influenzae, M.
catarrhalis, and E. coli.
7. The method of claim 1, wherein the patient has a bacterial
infection selected from the group consisting of respiratory tract
infections, otitis media, meningitis, endocarditis, skin and soft
tissue infections, and mastitis.
8. The method of claim 1, wherein the patient is a human or a
non-human mammal.
9. The method of claim 8, wherein the patient is human and the
compound is administered in an amount of a least about 50 to about
3000 mg per day.
10. The method of claim 9, wherein the patient is human and the
compound is administered in an amount of about 1500 mg per day.
11. The method of claim 9, wherein the patient is human and the
compound is administered in an amount of at least about 5 to about
20 mg per kg per day.
12. The method of claim 1, wherein the compound is
N-(4,5,-dibromothien-2-ylmethyl)-N'-(1H-imidazo[4,5-b]pyridine-2-yl)-prop-
ane-1,3-diamine.
13. The method of claim 1, wherein the compound is an
R-enantiomer.
14. A method of treating a resistant or multiply-resistant S.
aureus infection, the method comprising administering to a patient
in need thereof an antibacterially effective amount of a compound
of the formula (I): ##STR93## in which: R.sup.1 is an optionally
substituted aryl or an optionally substituted heteroaryl ring;
R.sup.2 is a 5 or 6-membered heteroaryl ring which is optionally
substituted with from 1 to 3 substituents selected from halo,
cyano, hydroxy, (C.sub.1-6)alkyl (optionally substituted by halo,
hydroxy, amino, mono to perfluoro(C.sub.1-3)alkyl, carboxy or
(C.sub.1-6)alkoxycarbonyl), (C.sub.3-7)cycloalkyl,
C.sub.(1-6)alkoxy, amino, mono- or di-(C.sub.1-6)alkylamino,
acylamino, carboxy, (C.sub.1-6)alkoxycarbonyl,
carboxy(C.sub.1-6)alkyloxy, (C.sub.1-6)alkylthio,
(C.sub.1-6)alkylsulphinyl, (C.sub.1-6)alkylsulphonyl, sulphamoyl,
mono- and di-(C.sub.1-6)alkylsulphamoyl, carbamoyl, mono- and
di-(C.sub.1-6)alkylcarbamoyl, and heterocyclyl; X is CH.sub.2 or
CHR.sup.3 in which R.sup.3 is C.sub.(1-6)alkyl or is linked to the
ortho position of an aryl or heteroaryl ring of R.sup.1 to form a 5
to 7 membered ring optionally including oxygen or nitrogen as a
ring atom; Y is C.sub.(1-3)alkylene or C.sub.(4-6)cycloalkylene;
including tautomeric forms of the imidazole ring; and salts thereof
and excluding 8-[2-(benzylamino)ethylamino]theophylline.
15. A method of treating a resistant or multiply-resistant E.
faecalis infection, the method comprising administering to a
patient in need thereof an antibacterially effective amount of a
compound of the formula (I): ##STR94## in which: R.sup.1 is an
optionally substituted aryl or an optionally substituted heteroaryl
ring; R.sup.2 is a 5 or 6-membered heteroaryl ring which is
optionally substituted with from 1 to 3 substituents selected from
halo, cyano, hydroxy, (C.sub.1-6)alkyl (optionally substituted by
halo, hydroxy, amino, mono to perfluoro(C.sub.1-3)alkyl, carboxy or
(C.sub.1-6)alkoxycarbonyl), (C.sub.3-7)cycloalkyl,
C.sub.(1-6)alkoxy, amino, mono- or di-(C.sub.1-6)alkylamino,
acylamino, carboxy, (C.sub.1-6)alkoxycarbonyl,
carboxy(C.sub.1-6)alkyloxy, (C.sub.1-6)alkylthio,
(C.sub.1-6)alkylsulphinyl, (C.sub.1-6)alkylsulphonyl, sulphamoyl,
mono- and di-(C.sub.1-6)alkylsulphamoyl, carbamoyl, mono- and
di-(C.sub.1-6)alkylcarbamoyl, and heterocyclyl; X is CH.sub.2 or
CHR.sup.3 in which R.sup.3 is C.sub.(1-6)alkyl or is linked to the
ortho position of an aryl or heteroaryl ring of R.sup.1 to form a 5
to 7 membered ring optionally including oxygen or nitrogen as a
ring atom; Y is C.sub.(1-3)alkylene or C.sub.(4-6)cycloalkylene;
including tautomeric forms of the imidazole ring; and salts thereof
and excluding 8-[2-(benzylamino)ethylamino]theophylline.
16. A method of treating a resistant or multiply-resistant S.
epidermidis infection, the method comprising administering to a
patient in need thereof an antibacterially effective amount of a
compound of the formula (I): ##STR95## in which: R.sup.1 is an
optionally substituted aryl or an optionally substituted heteroaryl
ring; R.sup.2 is a 5 or 6-membered heteroaryl ring which is
optionally substituted with from 1 to 3 substituents selected from
halo, cyano, hydroxy, (C.sub.1-6)alkyl (optionally substituted by
halo, hydroxy, amino, mono to perfluoro(C.sub.1-3)alkyl, carboxy or
(C.sub.1-6)alkoxycarbonyl), (C.sub.3-7)cycloalkyl,
C.sub.(1-6)alkoxy, amino, mono- or di-(C.sub.1-6)alkylamino,
acylamino, carboxy, (C.sub.1-6)alkoxycarbonyl,
carboxy(C.sub.1-6)alkyloxy, (C.sub.1-6)alkylthio,
(C.sub.1-6)alkylsulphinyl, (C.sub.1-6)alkylsulphonyl, sulphamoyl,
mono- and di-(C.sub.1-6)alkylsulphamoyl, carbamoyl, mono- and
di-(C.sub.1-6)alkylcarbamoyl, and heterocyclyl; X is CH.sub.2 or
CHR.sup.3 in which R.sup.3 is C.sub.(1-6)alkyl or is linked to the
ortho position of an aryl or heteroaryl ring of R.sup.1 to form a 5
to 7 membered ring optionally including oxygen or nitrogen as a
ring atom; Y is C.sub.(1-3)alkylene or C.sub.(4-6)cycloalkylene;
including tautomeric forms of the imidazole ring; and salts thereof
and excluding 8-[2-(benzylamino)ethylamino]theophylline.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 11/223,327, filed Sep. 9, 2005, now U.S. Pat. No. 7,220,757,
which is a continuation of U.S. application Ser. No. 10/729,416,
filed Dec. 5, 2003, now U.S. Pat. No. 6,943,175, which claims the
benefit under 35 U.S.C. .sctn. 119(b)(1) of United Kingdom patent
application serial number 0228545.0, filed Dec. 6, 2002.
TECHNICAL FIELD
[0002] The present invention relates to novel
2--NH-heteroarylimidazoles which are inhibitors of bacterial
methionyl t-RNA synthetase (MRS), processes for their preparation
and their use in therapy as anti-bacterial agents.
BACKGROUND OF THE INVENTION
[0003] t-RNA synthetases are involved in protein biosynthesis so
that inhibition thereof may be expected to lead to a cessation of
cell growth. Thus, for instance, the compound mupirocin, produced
by the organism Pseudomonas fluorescens, is an anti-bacterial agent
and is used as the active ingredient in the product Bactroban,
marketed by GlaxoSmithKline. Mupirocin has been shown to be an
inhibitor of the isoleucyl t-RNA synthetase. Each t-RNA synthetase
represents a separate target for drug discovery. t-RNA synthetase
inhibitors which are selective for bacterial cells over mammalian
cells are of considerable therapeutic interest as they have the
potential to be used as anti-bacterial agents.
[0004] The sequence of the t-RNA synthetase genes in the Gram
positive organism S. aureus have recently been determined (see, for
instance, European Patent application no 97300317.1, SmithKline
Beecham, for S. aureus MRS), thereby assisting the process of
identifying inhibitors. In addition, the sequence of t-RNA
synthetase genes in other pathogenic bacteria, for instance the
Gram negative organism H. influenzae, has also been published (R.
D. Fleischmann et al., Science, 269, 496-512, 1995).
[0005] Lespagnol et al have described a group of 8-substituted
theophylline derivatives, in particular
8-[2-(benzylamino)ethylamino]theophylline, which have hypotensive
activity (Ann Pharm Fr, 1968, 26(3), 207-14).
[0006] The present invention is directed toward overcoming one or
more of the problems discussed above.
DETAILED DESCRIPTION OF THE INVENTION
[0007] We have now found a novel class of 2--NH-substituted
heteroarylimidazoles which are potent inhibitors of bacterial
methionyl t-RNA synthetase. Accordingly, the present invention
provides a compound of the formula (I): ##STR1## in which:
[0008] R.sup.1 is an optionally substituted aryl or an optionally
substituted heteroaryl ring;
[0009] R.sup.2 is the residue of a 5 or 6-membered heteroaryl ring
which is optionally substituted with from 1 to 3 substituents
selected from halo, cyano, hydroxy, (C.sub.1-6)alkyl (optionally
substituted by halo, hydroxy, amino, mono to
perfluoro(C.sub.1-3)alkyl, carboxy or (C1-6)alkoxycarbonyl),
(C3-7)cycloalkyl, C(1-6)alkoxy, amino, mono- or
di-(C1-6)alkylamino, acylamino, carboxy, (C.sub.1-6)alkoxycarbonyl,
carboxy(C.sub.1-6)alkyloxy, (C.sub.1-6)alkylthio,
(C.sub.1-6)alkylsulphinyl, (C.sub.1-6)alkylsulphonyl, sulphamoyl,
mono- and di-(C.sub.1-6)alkylsulphamoyl, carbamoyl, mono- and
di-(C.sub.1-6)alkylcarbamoyl, and heterocyclyl;
[0010] X is CH.sub.2 or CHR.sup.3 in which R.sup.3 is
C.sub.(1-6)alkyl or is linked to the ortho position of an aryl or
heteroaryl ring of R.sup.1 to form a 5 to 7 membered ring
optionally including oxygen or nitrogen as a ring atom;
[0011] Y is C.sub.(1-3)alkylene or C.sub.(4-6)cycloalkylene;
[0012] including tautomeric forms of the imidazole ring; and
salts thereof, preferably pharmaceutically acceptable salts
thereof, and excluding
8-[2-(benzylamino)ethylamino]theophylline.
[0013] Compounds of formula (I) are inhibitors of bacterial
methionyl tRNA synthetase.
[0014] Representative examples of R.sup.1 when aryl include phenyl
and naphthyl, preferably phenyl, each of which may be optionally
substituted with up to three substituents. Representative examples
of such substituents include C.sub.(1-6) alkyl, C.sub.(1-6)
alkenyl, C.sub.(1-6) alkynyl, C.sub.(1-6) alkoxy, halo, cyano,
amino, sulphamoyl, phenylcarbonyl, aryl, and benzyloxy. Preferably,
the phenyl or naphthyl is substituted by two or three substituents
such as halo, C.sub.(1-6) alkyl, C.sub.(1-6) alkenyl, C.sub.(1-6)
alkynyl, C.sub.(1-6) alkoxy or C.sub.(1-6) alkylthio.
[0015] Representative examples of R.sup.1 when heteroaryl include
pyrrolyl, thienyl, furanyl, pyridyl, quinolinyl, benzofuranyl, and
indolyl, preferably thienyl or indolyl, each of which may be
optionally substituted with up to three substituents. Preferably,
the heteroaryl ring is substituted by two or substituents such as
halo, optionally substituted C.sub.(1-6) alkyl, optionally
substituted C.sub.(1-6) alkenyl, C.sub.(1-6) alkynyl, or
C.sub.(1-6) alkoxy. Representative examples of such substituents
include halo, mono to perfluoroC.sub.(1-6)alkyl and mono to
perfluoroC.sub.(1-6)alkenyl.
[0016] Preferred examples of aryl and heteroaryl groups for R.sup.1
include phenyl, indolyl and thienyl.
[0017] Representative heteroaryl rings formed by R.sup.2 are
nitrogen-containing heteroaryl rings, having 6 ring atoms and
including one or two nitrogen atoms, for instance b- or c-pyrido,
d-pyridimo or c-pyridazino; or sulfur-containing heteroaryl rings,
having 5 ring atoms, for instance c-thieno. Preferably, the
heteroaryl ring is unsubstituted. Preferably, the ring is
c-pyridazino.
[0018] Representative examples of X include CH.sub.2 or forming
with R.sup.1 a 5-7-membered ring fused to an aryl or heteroaryl
ring, preferably including oxygen or nitrogen as a ring atom, for
instance tetrahydroquinolinyl and chromanyl.
[0019] Representative examples of Y include a C.sub.2 alkylene
chain or a 1,2-cyclopentylene group.
[0020] Salts may be formed from inorganic and organic acids.
Representative examples of suitable inorganic and organic acids
from which pharmaceutically acceptable salts of compounds of
formula (I) may be formed include maleic, fumaric, benzoic,
ascorbic, pamoic, succinic, bismethylenesalicylic, methanesulfonic,
ethanedisulfonic, acetic, propionic, tartaric, salicylic, citric,
gluconic, aspartic, stearic, palmitic, itaconic, glycolic,
p-aminobenzoic, glutamic, benzenesulfonic, hydrochloric,
hydrobromic, sulfuric, cyclohexylsulfamic, phosphoric and nitric
acids.
[0021] When used herein, the term "alkyl" and similar terms such as
"alkoxy" includes all straight chain and branched isomers.
Representative examples thereof include methyl, ethyl, n-propyl,
iso-propyl, n-butyl, sec-butyl, iso-butyl, t-butyl, n-pentyl and
n-hexyl.
[0022] When used herein, the terms "alkenyl" and "alkynyl" include
all straight chain and branched isomers. Representative examples
thereof include vinyl, ethynyl and 1-propynyl.
[0023] Preferred substituents for alkyl and alkenyl groups include,
for example, and unless otherwise defined, halogen, cyano, azido,
nitro, carboxy, (C.sub.1-6)alkoxycarbonyl, carbamoyl, mono- or
di-(C.sub.1-6)alkylcarbamoyl, sulpho, sulphamoyl, mono- or
di-(C.sub.1-6)alkylsulphamoyl, amino, mono- or
di-(C.sub.1-6)alkylamino, acylamino, ureido,
(C.sub.1-6)alkoxycarbonylamino, 2,2,2-trichloroethoxycarbonylamino,
aryl, heterocyclyl, hydroxy, (C.sub.1-6)alkoxy, acyloxy, oxo, acyl,
2-thienoyl, (C.sub.1-6)alkylthio, (C.sub.1-6)alkylsulphinyl,
(C.sub.1-6)alkylsulphonyl, hydroxyimino, (C.sub.1-6)alkoxyimino,
hydrazino, hydrazono, benzohydroximoyl, guanidino, amidino and
iminoalkylamino.
[0024] When used herein, the term "aryl" includes, unless otherwise
defined, phenyl or naphthyl optionally substituted with up to five,
preferably up to three substituents.
[0025] When substituted, an aryl group may have up to three
substituents. Preferred substituents for an aryl group include, for
example, and unless otherwise defined, halogen, cyano,
(C.sub.1-6)alkyl, mono to perfluoro(C.sub.1-3)alkyl,
(C.sub.3-7)cycloalkyl, (C.sub.2-6)alkenyl, (C.sub.1-6)alkoxy,
(C.sub.2-6)alkenoxy, arylC.sub.(1-6)alkoxy, halo(C.sub.1-6)alkyl,
hydroxy, amino, mono- or di-(C.sub.1-6)alkylamino, acylamino,
nitro, carboxy, (C.sub.1-6)alkoxycarbonyl,
(C.sub.1-6)alkenyloxycarbonyl,
(C.sub.1-6)alkoxycarbonyl(C.sub.1-6)alkyl, carboxy(C.sub.1-6)alkyl,
(C.sub.1-6)alkylcarbonyloxy, carboxy(C.sub.1-6)alkyloxy,
(C.sub.1-6)alkoxycarbonyl(C.sub.1-6)alkoxy, (C.sub.1-6)alkylthio,
(C.sub.1-6)alkylsulphinyl, (C.sub.1-6)alkylsulphonyl, sulphamoyl,
mono- and di-(C.sub.1-6)-alkylsulphamoyl, carbamoyl, mono- and
di-(C.sub.1-6)alkylcarbamoyl, and heterocyclyl.
[0026] When used herein, the term "heteroaryl" includes single or
fused rings comprising up to four hetero-atoms in the ring selected
from oxygen, nitrogen and sulphur. Preferably the heteroaryl ring
comprises from 4 to 7, preferably 5 to 6, ring atoms. A fused
heteroaryl ring system may include carbocyclic rings and need only
include one heterocyclic ring.
[0027] When used herein, the term "heterocyclyl" includes aromatic
and non-aromatic single or fused rings comprising up to four
hetero-atoms in the ring selected from oxygen, nitrogen and
sulphur. Suitably the heterocyclic ring comprises from 4 to 7,
preferably 5 to 6, ring atoms. A fused heterocyclic ring system may
include carbocyclic rings and need only include one heterocyclic
ring.
[0028] When substituted, a heteroaryl or a heterocyclyl group may
have up to three substituents. Preferred such substituents include
those previously mentioned for an aryl group as well as oxo.
[0029] When used herein, the terms "halogen" and "halo" include
fluorine, chlorine, bromine and iodine and fluoro, chloro, bromo
and iodo, respectively.
[0030] The compounds according to the invention are suitably
provided in substantially pure form, for example at least 50% pure,
suitably at least 60% pure, advantageously at least 75% pure,
preferably at least 85% pure, more preferably at least 95% pure,
especially at least 98% pure, all percentages being calculated as
weight/weight. An impure or less pure form of a compound according
to the invention may, for example, be used in the preparation of a
more pure form of the same compound or of a related compound (for
example a corresponding derivative) suitable for pharmaceutical
use.
[0031] It will be appreciated that certain compounds of the present
invention may comprise one or more chiral centres so that compounds
may exist as stereoisomers, including diastereoisomers and
enantiomers. The present invention covers all such stereoisomers,
and mixtures thereof, including racemates.
[0032] Preferred compounds of formula (I) include the compounds of
Examples 2, 3, 6, 9, 11, 12, 15, 17, 22, 30, 32, 34-37, 39-41, and
43-44.
[0033] A compound of formula (I) may be prepared by reacting an
imidazole compound of formula (II): ##STR2## in which R.sup.2 is as
hereinbefore defined; and R.sup.4 is a leaving group such as halo,
for instance chloro, or C.sub.(1-6) alkylthio; with an amine of the
formula (III): R.sup.1XNHYCH.sub.2NH.sub.2 (III) in which R.sup.1,
X and Y are as hereinbefore defined; or an activated derivative
thereof; under nucleophilic displacement conditions.
[0034] Suitable conditions are well known in the art and include
the use of a large excess of the compound of formula (III) to drive
the reaction to completion and heating at a temperature of
60-130.degree. C. Addition of a base may be advantageous in some
cases, e.g. a tertiary base such as
N,N-di(cyclohexyl)ethylamine.
[0035] A compound of formula (I) may also be prepared by reacting a
compound of formula (IV): ##STR3## in which R.sup.2 and Y are as
hereinbefore defined; with either:
[0036] (a) for a compound of formula (I) in which X is CH.sub.2, an
aldehyde of formula (V): R.sup.1CHO (V) in which R.sup.1 is as
hereinbefore defined; under reductive alkylation conditions; or
[0037] (b) for a compound of formula (I) in which X is CHR.sup.3, a
ketone of formula (VI): R.sup.1R.sup.3CO (VI) in which R.sup.1 and
R.sup.3 are as hereinbefore defined; under reductive alkylation
conditions.
[0038] Suitable reductive alkylating conditions are well known in
the art and include for instance, the use of sodium
triacetoxyborohydride in a solvent system such as DMF/acetic acid
or sodium cyanoborohydride in methanol/acetic acid. Reductive
alkylation with an aldehyde is typically carried out at room
temperature for a period of 1-16 h. Reductive alkylation with a
ketone is typically carried out in refluxing methanol for a period
of 16-40 h.
[0039] A compound of formula (IV) may be prepared by reacting a
compound of formula (II) with a compound of formula (III) in which
R.sup.1X is hydrogen.
[0040] Alternatively, compounds of formula (I) in which Y is
C.sub.(1-3)alkylene may be prepared by a reductive amination
process in which the amine and aldehyde/ketone are reversed. Thus,
in a further aspect, the present invention provides for a process
for preparing a compound of formula (I) in which Y is
C.sub.(1-3)alkylene which process comprises reacting a compound of
formula (VII): ##STR4## in which R.sup.2 is as hereinbefore
defined, and Y.sup.1 is C.sub.(0-2)alkylene; with an amine of
formula (VIII): R.sup.1XNH.sub.2 (VIII) in which R.sup.1 and X are
as hereinbefore defined; under reductive alkylation conditions, as
hereinbefore described.
[0041] A compound of formula (VII) may be prepared in a similar way
to a compound of formula (IV), using a compound of formula (II) and
an amine of the formula
(C.sub.(1-6)alkoxyl).sub.2CHY.sup.1CH.sub.2NH.sub.2, followed by
acid hydrolysis to liberate the aldehyde from the acetal.
[0042] Compounds of formula (VIII) are amines and are either
commercially available or may be prepared form available starting
materials using methods well known in the art for preparing amines,
for instance by functional group interconversion.
[0043] The compounds of this invention are active against a range
of important pathogenic bacteria, including Gram positive
organisms, such as Staphylococci, for instance S. aureus Oxford and
coagulase negative strains of Staphylococci such as S. epidermidis;
Streptococci, for instance S. pyogenes CN10 and S. pneumoniae R6;
and Enterococci, for instance Ent. faecelis I. Preferably,
compounds of this invention are also active against Gram negative
organisms, such as Haemophilus, for instance H. influenzae Q1;
Moraxella, for instance M. catarrhalis 1502; and Escherichia, for
instance E. Coli DC0. The most preferred compounds of the present
invention will be active against the organisms S. aureus; S.
pneumoniae; Ent. faecelis; H. influenzae and M. catarrhalis.
[0044] In addition, compounds of this invention are active against
Staphylococci organisms such as S. aureus and coagulase negative
strains of Staphylocci such as S. epidermidis which are resistant
(including multiply-resistant) to other anti-bacterial agents, for
instance, .beta.-lactam antibiotics such as, for example,
methicillin; macrolides; aminoglycosides, and lincosamides.
Compounds of the present invention are therefore useful in the
treatment of MRSA and MRCNS.
[0045] Compounds of the present invention are also active against
strains of E. faecalis including vancomycin resistant strains and
therefore of use in treating infections associated with VRE
organisms. Furthermore, compounds of the present invention are
useful in the treatment of Staphylococci organisms which are
resistant to mupirocin.
[0046] Bacterial infections which may be treated include
respiratory tract infections, otitis media, meningitis,
endocarditis, skin and soft tissue infections in man, mastitis in
cattle, and also respiratory infections in farm animals such as
pigs and cattle. Accordingly, in a further aspect, the present
invention provides a method of treating bacterial infection in
human or non-human animals, which method comprises administering a
therapeutically effective amount of a compound of formula (I) as
hereinbefore defined, to a human or non-human animal in need of
such therapy. It will be appreciated that a compound of the present
invention which has a broad spectrum of anti-bacterial activity,
including activity against both Gram positive and Gram negative
bacteria will be of general use in the community for the empiric
treatment of community acquired infections. In comparison, a
compound of the present invention with a more limited spectrum, for
instance activity against Gram positive bacteria, is more likely to
be used in circumstances where the causative pathogenic organism
has been identified.
[0047] The present invention provides a pharmaceutical composition
comprising a compound of formula (I) together with a
pharmaceutically acceptable carrier or excipient.
[0048] The present invention also provides a method of treating
bacterial infections in animals, especially in humans and in
domesticated mammals, which comprises administering a compound of
formula (I), or a composition according to the invention, to a
patient in need thereof.
[0049] The invention further provides the use of a compound of
formula (I) in the preparation of a medicament composition for use
in the treatment of bacterial infections.
[0050] The compounds and compositions according to the invention
may be formulated for administration in any convenient way for use
in human or veterinary medicine, by analogy with other
antibiotics.
[0051] The compounds and compositions according to the invention
may be formulated for administration by any route, for example
oral, topical or parenteral. The compositions may, for example, be
made up in the form of tablets, capsules, powders, granules,
lozenges, creams, syrups, or liquid preparations, for example
solutions or suspensions, which may be formulated for oral use or
in sterile form for parenteral administration by injection or
infusion.
[0052] Tablets and capsules for oral administration may be in unit
dosage form, and may contain conventional excipients including, for
example, binding agents, for example, syrup, acacia, gelatin,
sorbitol, tragacanth, or polyvinylpyrrollidone; fillers, for
example lactose, sugar, maize-starch, calcium phosphate, sorbitol
or glycine; tabletting lubricants, for example magnesium stearate,
talc, polyethylene glycol or silica; disintegrants, for example
potato starch; and pharmaceutically acceptable wetting agents, for
example sodium lauryl sulphate. The tablets may be coated according
to methods well known in normal pharmaceutical practice.
[0053] Oral liquid preparations may be in the form of, for example,
aqueous or oily suspensions, solutions, emulsions, syrups or
elixirs, or may be presented as a dry product for reconstitution
with water or another suitable vehicle before use. Such liquid
preparations may contain conventional additives, including, for
example, suspending agents, for example sorbitol, methyl cellulose,
glucose syrup, gelatin, hydroxyethyl cellulose, carboxymethyl
cellulose, aluminium stearate gel or hydrogenated edible fats;
emulsifying agents, for example lecithin, sorbitan monooleate or
acacia; non-aqueous vehicles (which may include edible oils), for
example almond oil, oily esters (for example glycerine), propylene
glycol, or ethyl alcohol; preservatives, for example methyl or
propyl p-hydroxybenzoate or sorbic acid; and, if desired,
conventional flavouring and colour agents.
[0054] Compositions according to the invention intended for topical
administration may, for example, be in the form of ointments,
creams, lotions, eye ointments, eye drops, ear drops, impregnated
dressings, and aerosols, and may contain appropriate conventional
additives, including, for example, preservatives, solvents to
assist drug penetration, and emollients in ointments and creams.
Such topical formulations may also contain compatible conventional
carriers, for example cream or ointment bases, and ethanol or oleyl
alcohol for lotions. Such carriers may constitute from about 1% to
about 98% by weight of the formulation; more usually they will
constitute up to about 80% by weight of the formulation.
[0055] Compositions according to the invention may be formulated as
suppositories, which may contain conventional suppository bases,
for example cocoa-butter or other glycerides.
[0056] Compositions according to the invention intended for
parenteral administration may conveniently be in fluid unit dosage
forms, which may be prepared utilizing the compound and a sterile
vehicle, water being preferred. The compound, depending on the
vehicle and concentration used, may be either suspended or
dissolved in the vehicle. In preparing solutions, the compound may
be dissolved in water for injection and filter-sterilised before
being filled into a suitable vial or ampoule, which is then sealed.
Advantageously, conventional additives including, for example,
local anaesthetics, preservatives, and buffering agents can be
dissolved in the vehicle. In order to enhance the stability of the
solution, the composition may be frozen after being filled into the
vial, and the water removed under vacuum; the resulting dry
lyophilized powder may then be sealed in the vial and a
accompanying vial of water for injection may be supplied to
reconstitute the liquid prior to use. Parenteral suspensions may be
prepared in substantially the same manner except that the compound
is suspended in the vehicle instead of being dissolved and
sterilisation cannot be accomplished by filtration. The compound
may instead be sterilised by exposure to ethylene oxide before
being suspended in the sterile vehicle. Advantageously, a
surfactant or wetting agent is included in such suspensions in
order to facilitate uniform distribution of the compound.
[0057] A compound or composition according to the invention may
suitably be administered to the patient in an antibacterially
effective amount.
[0058] A composition according to the invention may suitably
contain from 0.1% by weight, preferably from 10 to 60% by weight,
of a compound according to the invention (based on the total weight
of the composition), depending on the method of administration.
[0059] The compounds according to the invention may suitably be
administered to the patient at a daily dosage of from 1.0 to 100
mg/kg of body weight. For an adult human (of approximately 70 kg
body weight), from 50 to 3000 mg, for example about 1500 mg, of a
compound according to the invention may be administered daily.
Suitably, the dosage for adult humans is from 5 to 20 mg/kg per
day. Higher or lower dosages may, however, be used in accordance
with normal clinical practice.
[0060] When the compositions according to the invention are
presented in unit dosage form, each unit dose may suitably comprise
from 25 to 1000 mg, preferable from 50 to 500 mg, of a compound
according to the invention.
EXAMPLES
[0061] The following examples are provided for illustrative
purposes only and are not intended to limit the scope of the
invention. TABLE-US-00001 ##STR5## Ex. R.sup.a R.sup.b 1 ##STR6##
##STR7## 2 ##STR8## ##STR9## 3, 11 (e- nan- tiom- er) ##STR10##
##STR11## 4 ##STR12## ##STR13## 5 ##STR14## ##STR15## 6 ##STR16##
##STR17## 7 ##STR18## ##STR19## 8 ##STR20## ##STR21## 9 ##STR22##
##STR23## 10 ##STR24## ##STR25## 12 ##STR26## ##STR27## 13
##STR28## ##STR29## 14 ##STR30## ##STR31## 15 ##STR32## ##STR33##
16 ##STR34## ##STR35## 17 ##STR36## ##STR37## 18 ##STR38##
##STR39## 19 ##STR40## ##STR41## 20 ##STR42## ##STR43## 21
##STR44## ##STR45## 22 ##STR46## ##STR47## 23 ##STR48## ##STR49##
24 ##STR50## ##STR51## 25 ##STR52## ##STR53## 26 ##STR54##
##STR55## 27 ##STR56## ##STR57## 28 ##STR58## ##STR59## 29
##STR60## ##STR61## 30 ##STR62## ##STR63## 31 ##STR64## ##STR65##
32 ##STR66## ##STR67## 33 ##STR68## ##STR69## 34 ##STR70##
##STR71## 35 ##STR72## ##STR73## 36 ##STR74## ##STR75## 37
##STR76## ##STR77## 38 ##STR78## ##STR79## 39 ##STR80## ##STR81##
40 ##STR82## ##STR83## 41 ##STR84## ##STR85## 42 ##STR86##
##STR87## 43 ##STR88## ##STR89## 44 ##STR90## ##STR91##
General Method for Reductive Amination
[0062] To a suspension of the amine (0.2 mmol) (containing 0.5 mmol
sodium acetate if the amine was present as the dihydrochloride) in
methanol (2 ml) was added the aldehyde (0.2 mmol) in methanol (2
ml) and acetic acid (0.033 ml). After stirring under argon for 10
min, NaCNBH.sub.3 (24 mg, 0.4 mmol) in MeOH (1 ml) was added and
the reaction stirred for 16 h. The reaction mixture was applied to
a 2 g Varian Bond Elute SCX cartridge which was flushed with MeOH
(8 ml). The cartridge was then eluted with 8 ml 0.2 M NH.sub.3 in
MeOH, and this eluate evaporated to dryness. The residue was
purified by chromatography on silica gel eluting with 2-10% (9:1
MeOH/20 M NH.sub.3) in CH.sub.2Cl.sub.2. Product-containing
fractions were combined and evaporated under reduced pressure to
give the product as a white solid. To convert this into the
corresponding dihydrochloride, the solid was dissolved in 1.0 M HCl
in methanol (0.4 ml) and the solution evaporated to dryness.
Example 1
N-(3,5-Dibromobenzyl)-N'-(1H-imidazo[4,5-b]pyridin-2-yl)-propane-1,3-diami-
ne
[0063] a) 1,3-Dihydroimidazo[4,5-b]pyridine-2-thione To
2,3-diaminopyridine (4.36 g, 40 mmol) in pyridine (40 ml) was added
carbon disulfide (3.6 ml, 60 mmol). The mixture was heated to
50.degree. C. for 6 h then concentrated to low volume by
evaporation under reduced pressure and the residue triturated with
tetrahydrofuran. The pale brown solid was collected by filtration
and dried to give a first crop of 3.6 g. A second crop (2.44 g) was
obtained from the filtrate by re-evaporation and trituration with
tetrahydrofuran. m/z (ESI+) 152 (MH.sup.+, 100%).
[0064] b) 2-Methanesulfanyl-1H-imidazo[4,5-b]pyridine To compound
1a (5.55 g, 36.75 mmol) in dry tetrahydrofuran (100 ml) under argon
was added triethylamine (5.66 ml, 40 mmol) and iodomethane (2.5 ml,
40 mmol). After stirring for 20 h at 20.degree. C. the solid was
removed by filtration and washed with THF. The combined filtrates
were evaporated to dryness and triturated with dichloromethane. The
solid was collected by filtration, (4.55 g, 75%). m/z (ESI+) 166
(MH.sup.+, 100%).
[0065] c)
N-(3,5-Dibromobenzyl)-N'-(1H-imidazo[4,5-b]pyridin-2-yl)-propan-
e-1,3-diamine Compound 1b (0.02 g) was treated with
N-(3,5-dibromobenzyl)propane-1,3-diamine (0.08 g) at 125.degree. C.
under argon for 24 h. The crude product was triturated with
methanol to give the title compound as a white solid (0.023 g, 50%)
m/z (ESI+) 438 (MH.sup.+, 100%).
Example 2
N-(4,6-Dichloro-1H-indol-2-ylmethyl)-N'-(H-imidazo[4,5-b]pyridin-2-yl)-pro-
pane-1,3-diamine
[0066] a) N-(1H-imidazo[4,5-b]pyridin-2-yl)propane-1,3-diamine
[0067] The product from 1b (4.55 g) was treated with
1,3-diaminopropane (40 ml) at reflux under argon for 50 h. The
solvent was removed by evaporation under reduced pressure and the
residue triturated with diethyl ether to give a brown solid. This
was purified by chromatography on silica gel eluting with 5-25%
(9:1 methanol/0.880 aq. ammonia) in dichloromethane to give the
required product, (2.6 g, 50%)
[0068] b)
N-(4,6-Dichloro-1H-indol-2-ylmethyl)-N'-(1H-imidazo[4,5-b]pyrid-
in-2-yl)-propane-1,3-diamine Compound 2a and
4,6-dichloroindole-2-carbaldehyde were allowed to react using the
general method for reductive amination on a 0.18 mmol scale to
yield the title compound, as a yellow foam (0.017 g, 24%).
.delta..sub.H (CD.sub.3OD) 7.95 (1H, dd, J=5.2, 1.5 Hz), 7.45 (1H,
dd, J=7.7, 1.5 Hz), 7.25 (1H, bs), 7.0 (1H, bs), 6.95(1H, dd,
J=7.7, 5.3 Hz), 6.45 (1H, bs), 3.9 (2H, s), 3.5 (2H, t, J=6.7 Hz),
2.75 (2H, t, J=6.9 Hz), 1.9 (2H, m); m/z (ES+) 389 (26%, MH.sup.+),
192 (100%).
Example 3
N-(6,8-Dibromo-1,2,3,4,-tetrahydroquinolin-4-yl)-N'-(1H-imidazo[4,5-b]pyri-
din-2-yl)-propane-1,3-diamine dihydrochloride
[0069] To N-(1H-imidazo[4,5-b]pyridin-2-yl)propane-1,3-diamine
(0.055 g, 0.29 mmol) and
6,8-dibromo-2,3,4,5-tetrahydroquinolin-4-one (0.088 g, 0.29 mmol)
in methanol (2 ml) and acetic acid (0.06 g) was added sodium
cyanoborohydride (0.019 g, 0.3 mmol). The reaction was then
refluxed for 20 h. The reaction mixture was applied to a 2 g SCX
cartridge which was flushed with MeOH (15 ml). The cartridge was
then eluted with 15 ml 0.2 M NH.sub.3 in MeOH, and this eluate
evaporated to dryness. Further purification on silica gel eluting
with 0-10% (9:1 methanol/0.880 aq. ammonia) in dichloromethane gave
the title compound, which was converted to its dihydrochloride by
dissolution in 1.0 M HCl in methanol (0.4 ml) and the solution
evaporated to dryness to give a white solid (0.060 g, 37%);
.delta..sub.H (CD.sub.3OD) 8.0 (1H, dd, J=6.3, 1.2 Hz), 7.9 (1H,
dd, J=6.5, 1.2 Hz), 7.55 (1H, d, J=2.2 Hz), 7.4 (1H, d, J=2.2 Hz),
7.25 (1H, dd, J=6.5, 6.3 Hz), 4.5 (1H, bs), 3.7 (2H, t, J=6.6 Hz),
3.65-3.1 (4H, m), 2.4 (1H, m), 2.2-1.95 (3H, m); m/z (ES+) 479 (6%,
MH.sup.+), 192 (100%).
Example 4
N-(4,5-Dibromothien-2-ylmethyl)-N'-(1H-imidazo[4,5-b]pyridin-2-yl)-propane-
-1,3-diamine
[0070] Using the general method for reductive amination on a 0.2
mmol scale 4,5-dibromothiophene-2-carbaldehyde was reacted with
compound 2a to give the title compound as the free base, 0.037 g,
40%. m/z (ES+) 444/446/448 (28/53/30%, MH.sup.+), 192 (100%).
Example 5
N-(3,5-Dibromobenzyl)-N'-(H-imidazo[4,5-c]pyridin-2-yl)-propane-1,3-diamin-
e
[0071] a) 1,3-Dihydroimidazo[4,5-c]pyridine-2-thione To
3,4-diaminopyridine (4.66 g, 42.75 mmol) in pyridine (40 ml) was
added carbon disulfide (4 ml, 60 mmol). The mixture was heated to
60oC for 24 h then concentrated to low volume by evaporation under
reduced pressure and the residue triturated with dry
tetrahydrofuran. The off white solid was collected by filtration
and dried to give a first crop of 3.87 g. A second crop (0.48 g)
was obtained from the filtrate (total 4.35 g, 67%). m/z (ESI+) 152
(MH+, 100%).
[0072] b) 2-Methanesulfanyl-1H-imidazo[4,5-c]pyridine To compound
5a (0.38 g, 2.5 mmol) in dry DMF (10 ml) under argon was added
triethylamine (0.38 ml) and iodomethane (0.163 ml). After stirring
at 20.degree. C. for 1 h the solution was concentrated by
evaporation under reduced pressure and the residue triturated with
diethyl ether. Purification on silica gel eluting with 0-12% (9:1
methanol/0.880 aq. ammonia) in dichloromethane gave the title
compound as a white solid, 0.16 g, 39%. m/z (ESI+) 166 (MH.sup.+,
100%).
[0073] c)
N-(3,5-dibromobenzyl)-N'-(1H-imidazo[4,5-c]pyridin-2-yl)-propan-
e-1,3-diamine
[0074] Compound 5b (0.035 g, 0.21 mmol) was treated with
N-(3,5-dibromobenzyl)propane-1,3-diamine (0.14 g) at 140.degree. C.
under argon for 24 h. The crude product was purified by
chromatography on silica gel eluting with 0-12% (9:1 methanol/0.880
aq. ammonia) in dichloromethane to give the title compound (0.055
g, 60%). m/z (ESI+) 438 (MH.sup.+, 100%).
Example 6
N-(4,6-Dichloro-1H-indol-2-ylmethyl)-N'-(1H-imidazo[4,5-b]pyrazin-2-yl)-pr-
opane-1,3-diamine dihydrochloride
[0075] a) 2-Methanesulfanyl-1H-imidazo[4,5-b]pyrazine. To a
solution of 1,3-dihydroimidazo[4,5-b]pyrazine-2-thione (Aust. J.
Chem. 1982, 35, 2299-2306, 0.86 g) in aqueous NaOH (2 M, 6 ml) was
added methyl iodide (0.35 ml). The resulting brown solution was
stirred for 20 minutes under argon at ambient temperature. The
reaction mixture was then diluted further with aqueous NaOH (2 M, 5
ml) and extracted with a small amount of CHCl.sub.3. The aqueous
layer was evaporated, dissolved in the minimum volume of aqueous
NaOH (2 M), and acidified to pH 6.5 with HCl (conc.). The solution
was then saturated with NaCl and left to crystallise for 18 h. The
crystals thus obtained were collected by filtration and dried under
vacuum to yield the title compound as brown crystals (0.50 g);
.delta..sub.H [(CD.sub.3).sub.2SO] 2.75 (s, 3H, SCH.sub.3), 8.18
(br, s, 1H, Ar--H), 8.30 (br, s, 1H, Ar--H), 13.54 (br, s, 1H, NH);
m/z (ESI+) 167 (MH.sup.+, 100%).
[0076] b) N-(1H-Imidazo[4,5-b]pyrazin-2-yl)-propane-1,3-diamine.
2-Methanesulfanyl-1H-imidazo[4,5-b]pyrazine (0.49 g) and
1,3-diaminopropane (2.5 ml) were heated under reflux for 18 h to
give a green solution which on cooling yielded a brown oil. The
excess diaminopropane was evaporated and the residue applied to a
column of silica gel eluting with 10%-15%-20% (MeOH:NH.sub.3 10:1)
in CH.sub.2Cl.sub.2 to yield the title compound (0.23 g) as a white
solid; .delta..sub.H (CD.sub.3OD/DCl) 2.16 (m, 2H,
CH.sub.2CH.sub.2CH.sub.2), 3.12 (m, 2H, NH.sub.2CH.sub.2), 3.69 (m,
2H, ArNHCH.sub.2), 8.11 (s, 2H, ArH); m/z (ESI+) 193 (MH.sup.+,
100%).
[0077] c)
N-(4,6-Dichloro-1H-indol-2-ylmethyl)-N'-(1H-imidazo[4,5-b]pyraz-
in-2-yl)-propane-1,3-diamine dihydrochloride.
N-(1H-Imidazo[4,5-b]pyrazin-2-yl)-propane-1,3-diamine and
4,6-dichloro-1H-indole-2-carbaldehyde were allowed to react using
the general method for reductive amination on a 0.2 mmol scale
followed by hydrochloride salt formation to yield the title
compound (0.017 g) as a white solid; .delta..sub.H (CD.sub.3OD/DCl)
2.21 (m, 2H, CH.sub.2CH.sub.2CH.sub.2), 3.27 (m, 2H,
ArCH.sub.2NHCH.sub.2), 3.69 (m, 2H, ArNHCH.sub.2), 4.48 (s, 2H,
ArCH.sub.2), 6.80 (m, 1H, ArH), 7.05 (m, 1H, ArH), 7.41 (m, 1H,
ArH), 8.10 (s, 2H, ArH); m/z (ESI+) 390 (MH.sup.+, 15%), 176
(100%).
Example 7
N-(4,6-Dichloro-1H-indol-2-ylmethyl)-N'-(9H-purin-8-yl)-propane-1,3-diamin-
e dihydrochloride
[0078] a) 7,9-Dihydropurine-8-thione. 4,5-Diaminopyrimidine (2.0 g)
was dissolved in pyridine (150 ml) containing NaOH (1.1 g) and to
this solution was added carbon disulphide (2.2 ml). The reaction
mixture was heated to 50.degree. C. for 18 h. Volatiles were
evaporated, the residue dissolved in H.sub.2O and acidified with
HCl (conc.). The acidic mixture was warmed on a water bath and NaOH
(conc.) added until precipitation was observed. This precipitate
was removed by filtration. On standing at room temperature yellow
crystals formed in the aqueous mother liquor which were filtered
off and dried under vacuum to give the title compound (1.20 g);
.delta..sub.H [(CD.sub.3).sub.2SO] 8.45 (s, 1H, ArH), 8.69 (s, 1H,
ArH), 13.02 (br, s, 1H, NH), 13.54 (br, s, 1H, NH).
[0079] b) 8-Methanesulfanyl-9H-purine: To a solution of
7,9-dihydropurine-8-thione (1.2 g) in aqueous NaOH (2 M, 8 ml) was
added iodomethane (0.5 ml). The resulting brown solution was
stirred for 90 minutes under argon at ambient temperature. The
reaction mixture was then diluted further with aqueous NaOH (2 M, 5
ml) and extracted with CHCl.sub.3 (10 ml). The aqueous layer was
separated and acidified to pH 6.0 with HCl (conc.). The white
precipitate thus obtained was collected by filtration and dried
under vacuum to yield the title compound as a white solid (0.65 g);
.delta..sub.H [(CD.sub.3).sub.2SO] 2.74 (s, 3H, SCH.sub.3), 8.75
(br, s, 1H, Ar--H), 8.85 (br, s, 1H, Ar--H), 13.48 (br, s, 1H, NH);
m/z (ESI+) 167 (MH.sup.+, 100%).
[0080] c) N-(9H-purin-8-yl)-propane-1,3-diamine:
8-Methanesulfanyl-9H-purine (0.65 g) and 1,3-diaminopropane (3.5
ml) were heated together under reflux for 6 h. The excess
diaminopropane was then evaporated and the residue repeatedly
chromatograph on silica gel eluting with (MeOH:NH.sub.3 10:1) in
CH.sub.2Cl.sub.2 to yield the title compound (0.24 g) as a white
solid; .delta..sub.H (CD.sub.3OD) 1.79 (m, 2H,
CH.sub.2CH.sub.2CH.sub.2), 2.78 (m, 2H, NH.sub.2CH.sub.2), 3.43 (m,
2H, ArNHCH.sub.2), 8.11 (s, 1H, ArH), 8.29 (s, 1H, ArH); m/z (ESI+)
193 (MH.sup.+, 100%).
[0081] d)
N-(4,6-Dichloro-1H-indol-2-ylmethyl)-N'-(9H-purin-8-yl)-propane-
-1,3-diamine dihydrochloride:
N.sup.1-(9H-purin-8-yl)-propane-1,3-diamine and
4,6-dichloro-1H-indole-2-carbaldehyde were allowed to react using
the general method for reductive amination on a 0.2 mmol scale
followed by hydrochloride salt formation to yield the title
compound (0.014 g) as a white solid; .delta..sub.H (CD.sub.3OD) of
corresponding free base 1.81 (m, 2H, CH.sub.2CH.sub.2CH.sub.2),
2.71 (m, 2H, ArCH.sub.2NHCH.sub.2), 3.44 (m, 2H, ArNHCH.sub.2),
3.89 (s, 2H, ArCH.sub.2), 6.38 (m, 1H, ArH), 6.91 (m, 1H, ArH),
7.18 (m, 1H, ArH), 8.12 (m, 1H, ArH), 8.42 (m, 1H, ArH); m/z (ESI+)
390 (MH.sup.+, 15%), 176 (100%).
Example 8
N-(4,5-Dibromothien-2-ylmethyl)-N'-(9H-purin-8-yl)-propane-1,3-diamine
dihydrochloride
[0082] N-(9H-purin-8-yl)-propane-1,3-diamine and
4,5-dibromothiophene-2-carbaldehyde were allowed to react using the
general method for reductive amination on a 0.2 mmol scale followed
by hydrochloride salt formation to yield the title compound (0.014
g) as a white solid; .delta..sub.H (CD.sub.3OD) 2.19 (m, 2H,
CH.sub.2CH.sub.2CH.sub.2), 3.23 (m, 2H, ArCH.sub.2NHCH.sub.2), 3.72
(m, 2H, ArNHCH.sub.2), 4.45 (s, 2H, ArCH.sub.2) 7.35 (br, s, 1H,
ArH), 8.56 (m, 1H, ArH), 8.95 (m, 1H, ArH); m/z (ESI+) 445
(MH.sup.+, 100%).
Example 9
N-(3-Bromo-5-methoxy-1H-indol-7-ylmethyl)-N'-(H-imidazo[4,5-b]pyridin-2-yl-
)-propane-1,3-diamine
[0083] a) 5-Methoxyindoline-7-carbaldehyde.
1-(tert-Butoxycarbonyl)-5-methoxyindoline (Heterocycles, 1992, 34,
1031; 1.75 g 7.0 mmol) was dissolved in dry THF, treated with TMEDA
(1.4 ml) and cooled to -78.degree. C. under an argon atmosphere. A
solution of s-butyl lithium (1.3 M in cyclohexane, 5.18 ml) was
added dropwise. After stirring at -78.degree. C. for 1 h, the
solution was treated with dry DMF (1.08 ml, 14 mmol) and stirred
for a further 0.5 h. The cooling bath was then removed and the
solution allowed to reach room temperature over 1 h. The reaction
mixture was quenched with 10% aqueous NH.sub.4Cl and the product
extracted into ethyl acetate. The extracts were combined, washed
with water and brine, dried (MgSO.sub.4) and evaporated. The
residue was chromatographed on Kieselgel 60 eluting with 0-20%
ethyl acetate in hexane. Product-containing fractions were combined
and evaporated to afford the title compound (510 mg); contaminated
with 35% (by weight) of the corresponding N-Boc analogue;
.delta..sub.H (CDCl.sub.3, inter alia) 3.03 (2H, t, J=8.0 Hz,
CH.sub.2), 3.76 (2H, t, J=8.1 Hz, CH.sub.2NH), 3.77 (3H, s, OMe),
6.42 (1H, br.s, NH), 6.73 (1H, d, J=0.8 Hz Ar--H), 6.90-6.92 (1H,
m, Ar--H), 9.79 (1H, s, CHO).
[0084] b) 5-Methoxyindole-7-carbaldehyde. The product from 9a (80
mg; containing 0.3 mmol 5-methoxyindoline-7-carbaldehyde) was
dissolved in dichloromethane (10 ml) and treated with MnO.sub.2
(344 mg, 4.0 mmol). The reaction mixture was stirred at room
temperature for 16 h, filtered through Celite and the solvent
removed in vacuo. The residue was chromatographed on Kieselgel 60
eluting with 0-20% ethyl acetate in hexane to afford the title
compound as a pale yellow solid (23 mg, 44%), .delta..sub.H
(CDCl.sub.3) 3.91(3H, s, OMe), 6.56 (1H, dd, J=2.2, 3.2 Hz, 3-H),
7.28 (1H, d, J=2.3 Hz, Ar--H), 7.33(1H, t, J=2.6 Hz, 2-H), 7.46(1H,
m, Ar--H), 9.93(1H, br.s, NH), 10.07 (1H, s, CHO).
[0085] c) 3-Bromo-5-methoxyindole-7-carbaldehyde. The product from
9b (40 mg, 0.22 mmol) was dissolved in dichloromethane (5 ml),
treated with N-bromosuccinimide (40 mg), and the mixture stirred at
room temperature for 16 h. The solution was then diluted with
dichloromethane, washed with water and brine, dried (MgSO.sub.4)
and evaporated. The residue was chromatographed on Kieselgel 60
eluting with 0-50% ethyl acetate in hexane. Product-containing
fractions were combined and evaporated to afford the title compound
as a pale pink solid (53 mg, 95%); .delta..sub.H (CDCl.sub.3) 3.94
(3H, s, OMe), 7.34 (3H, s, 2-H, 4-H, 6-H), 9.93(1H, br.s. NH),
10.06 (1H, s, CHO).
[0086] d)
N-(3-Bromo-5-methoxy-1H-indol-7-ylmethyl)-N'-(1H-imidazo[4,5-b]-
pyridin-2-yl)-propane-1,3-diamine. The product from 9c was coupled
compound 2a on a 2 mmol scale using the general method for
reductive amination to give the title compound as a white solid (11
mg, 13%); .delta..sub.H (CD.sub.3OD) 1.89 (2H, t, J=6.8 Hz,
CH.sub.2), 2.75 (2H, t, J=6.8 Hz, CH.sub.2), 3.47 (2H, t, J=6.6 Hz,
CH.sub.2), 3.82 (3H, s, OMe), 4.01 (2H, s, ArCH.sub.2), 6.84 (2H,
s, 2.times. indole-H), 6.93-6.97 (1H, m, pyr-H), 7.21 (1H, s,
indole-H), 7.41 (1H, d, J=7.7 Hz, pyr-H), 7.93 (1H, d, J=5.0 Hz,
pyr-H); m/z (CI.sup.+) 429 (MH.sup.+, 100%).
Example 10
N-(6-Ethyl-8-iodo-1,2,3,4-tetrahydroquinolin-4-yl)-N'-(H-imidazo[4,5-b]pyr-
idin-2-yl)-propane-1,3-diamine dihydrochloride
[0087] (a) 3-(4-Ethylphenylamino)propionic acid. To a solution of
4-ethylaniline (10 mmol) in acetonitrile (15 mL) at reflux was
added 2-oxetanone (10 mmol) over a 20 minute period. After 3 h the
mixture was cooled and the acetonitrile was evaporated. The residue
was recrystallised from toluene to yield the title compound as a
white solid; MS (ES.sup.-) 385(100%)[2M-H].sup.-,
192(76%)[M-H].sup.-.
[0088] (b) 6-Ethyl-2,3-dihydro-1H-quinolin-4-one. To phosphorus
pentoxide (15 g) was added 85% orthophosphoric acid (6 mL) and the
resultant mixture heated at 100.degree. C. for 0.5 h.
3-(4-Ethylphenylamino)propionic acid (1.8 mmol) was then added.
After 2 h the mixture was quenched with ice and stirred vigorously
for 0.5 h. The mixture was treated with aqueous ammonia solution
(pH 9.0), extracted with the ethyl acetate, dried and evaporated to
give the title compound; .delta..sub.H (CDCl.sub.3) 77.68(1H, d),
7.16(1H, d.times.d), 6.61(1H, d), 4.28(1H, br. s), 3.55(2H, t),
2.68(2H, t) 2.55(2H, q) 1.19(3H, t); MS (ES.sup.-)
174(30%)[M-H].sup.-.
[0089] (c)
N-(6-Ethyl-8-iodo-1,2,3,4-tetrahydroquinolin-4-yl)-N'-(1H-imidazo[4,5-b]p-
yridin-2-yl)-propane-1,3-diamine dihydrochloride.
6-Ethyl-8-iodo-1,2,3,4-tetrahydroquinol-4-one (140 mg, 0.47 mmol)
and N-(1H-imidazo[4,5-b]pyridin-2-yl)-propane-1,3-diamine (90 mg,
0.47 mmol) were allowed to react in an identical way to Example 3
and after purification and conversion to the dihydrochloride the
title compound was obtained as a white solid; .delta..sub.H
(CD.sub.3OD) 1.18 (3H, t), 2.07-2.16 (2H, m), 2.35-2.47 (3H, m),
3.21-3.43 (4H, m), 3.66 (2H, t), 4.42 (1H, br. s), 7.08 (1H, d),
7.26 (1H, d), 7.54 (1H, d), 7.88 (1H, dd), 7.98 (1H, dd); m/z
(CI.sup.+) 477 (MH.sup.+, 18%).
Example 11
N-(6,8-Dibromo-1,2,3,4-tetrahydroquinolin-4-yl)-N'-(1H-imidazo[4,5-b]pyrid-
in-2-yl)-propane-1,3-diamine dihydrochloride--Enantiomer B
[0090] (a) 6,8-Dibromo-1,2,3,4-tetrahydro-1H-quinolin-4-one
O-methyloxime. A mixture of
6,8-dibromo-1,2,3,4-tetrahydroquinolin-4-one (2 mmol) in ethanol (5
mL) was added to solution of O-methylhydroxylamine hydrochloride
(2.2 mmol) and sodium acetate (2.2 mmol) in water (10 mL). The
resultant mixture was heated at reflux for 18 h, cooled, the
solvent evaporated, and the residue partitioned between ethyl
acetate and water. The organic layer was separated, dried, and
evaporated to yield the title compound as pale yellow solid. m/z
(ES.sup.+) 374 (100%) [M+CH.sub.3CN]H.sup.+, 333 (80%)
MH.sup.+.
[0091] (b) 4-Amino-6,8-dibromo-1,2,3,4-tetrahydroquinoline. To a
solution of compound 11a (1.7 mmol) in THF (20 mL) at 5.degree. C.
under an atmosphere of argon was added portionwise, with rapid
stirring, zirconium tetrachloride (2.6 mmol). After 5 minutes a 2M
solution of lithium borohydride (2.6 mL) in THF was added dropwise
and the resultant solution was stirred at room temperature for 48
h. The mixture was treated dropwise with methanol (10 mL) and the
solvent evaporated. The residue was partitioned between 2M
hydrochloric acid and diethyl ether. The aqueous layer was
separated, made basic by the addition of conc. ammonium hydroxide
and extracted with dichloromethane. The resultant organic layer was
dried and evaporated to yield the title compound as white solid.
m/z (ES.sup.+) 305 (2%) MH.sup.+, 288 (100%)
[M-NH.sub.3]H.sup.+.
[0092] (c)
(2S)-N-(6,8-Dibromo-1,2,3,4-tetrahydroquinolin-4-yl)-2-methoxy-2-phenylac-
etamide--Diastereoisomer B. To a solution of compound 11b (0.6
mmol), (S)-.alpha.-methoxyphenylacetic acid (0.7 mmol), DEC (0.7
mmol), and HOAt (0.7 mmol) in dry DMF was added N-methylmorpholine
(0.7 mmol). After 16 h the solvent was evaporated and the residue
partitioned between 10% citric acid solution and ethyl acetate. The
organic layer was separated, washed with sodium bicarbonate
solution, dried and evaporated to yield a mixture of
diastereoisomeric amides as a pale yellow solid. Chromatography
over silica gel eluting with petroleum ether containing ethyl
acetate (30%) and collecting the slower running diastereoisomer,
gave the title compound as an oil which crystallised on standing to
form a white solid. .delta..sub.H (CDCl.sub.3) 2.01-2.10 (2H, m),
3.34 (3H, s), 3.43-3.50 (2H, m), 4.59 (1H, s), 4.66 (1H, s),
5.08-5.13 (1H, m), 6.91 (1H, d), 7.00 (1H, d), 7.26-7.43 (6H, m);
m/z (ES.sup.+) 453 (100%) MH.sup.+.
[0093] (d)
4-Amino-6,8-dibromo-1,2,3,4-tetrahydroquinoline--Enantiomer B.
Compound 11c (0.3 mmol) was dissolved in dioxane (3 mL) and 8M
hydrochloric acid. After heating under reflux for 5 h the mixture
was cooled and the solvent evaporated. The residue was partitioned
between water/ethyl acetate and the aqueous layer separated, made
basic by the addition of conc. ammonium hydroxide and extracted
with dichloromethane. Drying and evaporation gave the title
compound as an oil m/z. (ES.sup.+) 305 (2%) MH.sup.+, 288 (100%)
[M-NH.sub.3]H.sup.+.
[0094] (e)
N-(1H-imidazo[4,5-b]pyridin-2-yl)-(3,3-diethoxyprop-1-yl)amine--A
mixture of 3-amino-1,1-diethoxypropane (18 mmol) and compound 1b
(3.3 mmol) was heated at 130.degree. C. under argon. After 15 h the
mixture was cooled and triturated with cold diethyl ether the
resultant solid was filtered off and dried to yield the title
compound as a cream solid. m/z (CI.sup.+) 529 (5%) [2M]H.sup.+, 265
(100%) MH.sup.+
[0095] (f) 3-(1H-Imidazo[4,5-b]pyridin-2-ylamino)propionaldehyde
hydrochloride--Compound 11e (2 mmol) was dissolved in 1M
hydrochloric acid, warmed at 100.degree. C. for 15 minutes, cooled,
and the solvent evaporated to yield the title compound as a white
solid.
[0096] (g)
N-(6,8-Dibromo-1,2,3,4-tetrahydroquinolin-4-yl)-N'-(1H-imidazo[4,5-b]pyri-
din-2-yl)-propane-1,3-diamine dihydrochloride--Enantiomer B. A
mixture of compound 11f (0.13 mmol), compound 11d (0.13 mmol) and
sodium acetate (0.13 mmol) was dissolved in 1% acetic acid in
methanol (3 mL) for 2 h at room temperature. Sodium cyanborohydride
(0.13 mmol) was added and after 20 h the solvent was evaporated and
the mixture chromatographed over silica gel eluting with
dichloromethane containing 8% of 1:9 ammonium hydroxide/methanol
mixture. The resultant free base was converted to the title
compound as a white solid; .delta..sub.H (CD.sub.3OD) 8.0 (1H, dd,
J=6.3, 1.2 Hz), 7.9 (1H, dd, J=6.5, 1.2 Hz), 7.55 (1H, d, J=2.2
Hz), 7.4 (1H, d, J=2.2 Hz), 7.25 (1H, dd, J=6.5, 6.3 Hz), 4.5 (1H,
bs), 3.7 (2H, t, J=6.6 Hz), 3.65-3.1 (4H, m), 2.4 (1H, m), 2.2-1.95
(3H, m), m/z (ES.sup.+) 479 (6%, MH.sup.+), ee=99% (capillary zone
electrophoresis). Enantiomer B is the slower running enantiomer (by
CZE on fused silica 50 cm.times.50 micron id, voltage 20 KV, buffer
100 nM sodium phosphate containing 40 nM cyclodextrin)
[0097] (h)
N-(6,8-Dibromo-1,2,3,4-tetrahydroquinolin-4-yl)-N'-(1H-imidazo[4,5-b]pyri-
din-2-yl)-propane-1,3-diamine dihydrochloride--Enantiomer A.
Diastereomer A of
(2S)-N-(6,8-Dibromo-1,2,3,4-tetrahydroquinolin-4-yl)-2-methoxy-2-phe-
nylacetamide was recovered via chromatography in step (c) and used
to prepare Enantiomer A by a procedure analogous to steps
(d)-(g).
[0098] (i) Assignment of absolute configuration of isomers of step
(c). Colourless needle, 0.38.times.0.05.times.0.04 mm,
orthorhombic, space group P2.sub.12.sub.12.sub.1 (#19), T=150 K,
a=4.8880(4) .ANG., b=13.7295(12) .ANG., c=26.441(2) .ANG.,
V=1774.4(3) .ANG..sup.3, Z=4, D.sub.calc=1.700 Mgm.sup.-3,
F(000)=904, .mu.(Cu K.sub..alpha., .lamda.=1.54178 .ANG.)=5.902
mm.sup.-1, Bruker SMART 6000 diffractometer, 11989 reflections
collected (6.68.degree..ltoreq.2.theta..ltoreq.145.52.degree.),
3430 unique reflections (R.sub.int=0.0530), Gaussian absorption
correction (transmission=0.37514-0.80284), full-matrix
least-squares refinement (on F.sup.2) of 226 variables, R1=0.0316
(wR2=0.0789) for 3284 observed data with I.gtoreq.2.sigma.(I),
R1=0.0327 (wR2=0.0805) for all data, S=1.037,
w=1/[.sigma..sup.2(F.sub.o.sup.2)+(0.0604P).sup.2] where
P=[Max(F.sub.o.sup.2,0)+2F.sub.c.sup.2]/3, residual electron
density between -0.471 e.ANG..sup.-3 and 0.942 e.ANG..sup.-3,
absolute structure parameter=-0.046(19).
[0099] (j). Determination of enantiomeric excess (ee) of the final
products. The enantiomeric excess (ee) of the final products was
determined by chiral capillary zone electrophoresis (cze) to be
98.4% for the (R)-isomer and 99.0% for the (S)-isomer. (Fused
silica 50 cm.times.50 micron id, 20 kV, 100 mM sodium phosphate
buffer pH 2.5 containing 40 mM .alpha.-cyclodextrin).
Example 12
N-(3-Chloro-5-methoxy-1H-indol-7-ylmethyl)-N'-(1H-imidazo[4,5-b]pyridin-2--
yl)-propane-1,3-diamine
[0100] a) 3-Chloro-5-methoxy-1H-indol-7-carbaldehyde.
5-Methoxyindole-7-carbaldehyde 9b (40 mg, 0.22 mmol) was dissolved
in dichloromethane (5 ml), treated with N-chlorosuccinimide (40
mg), and the mixture stirred at room temperature for 16 h. The
solution was then diluted with dichloromethane, washed with water
and brine, dried (MgSO.sub.4) and evaporated to a pale brown
solid.
[0101] b)
N-(3-Chloro-5-methoxy-1H-indol-7-ylmethyl)-N'-(1H-imidazo[4,5-b-
]pyridin-2-yl)-propane-1,3-diamine. The product 12a was coupled to
compound 2a on a 0.2 mmol scale using the general method for
reductive amination to give the title compound as a white solid (7
mg, 9%); m/z (CI.sup.+) 386 (MH.sup.+, 70%).
Example 13
N-(4,5-Dibromo-3-methylthiophen-2-ylmethyl)-N'-(1H-imidazo[4,5-c]pyridin-2-
-yl)propane-1,3-diamine
[0102] a) N-(1H-imidazo[4,5-c]pyridin-2-yl)propane-1,3-diamine. A
mixture of compound 5b (1.97 g, 10 mmol) and 1,3-diaminopropane was
heated at 125.degree. C. for 24 h. The resultant mixture was cooled
and the excess reagent evaporated. The residue was chromatographed
over silica gel eluting with dichloromethane/methanol/ammonium
hydroxide (85:13.5:1.5) to yield the title compound. .delta..sub.H
(CD.sub.3OD) 8.31 (1H, s); 8.04 (1H, d); 7.25 (1H, d); 3.48 (2H,
t); 2.99 (2H, t); 1.97 (2H, m).
[0103]
N-(4,5-Dibromo-3-methylthiophen-2-ylmethyl)-N'-(1H-imidazo[4,5-c]p-
yridin-2-yl)propane-1,3-diamine. Using the general method for
reductive amination a mixture of
N-(3H-imidazo[4,5-c]pyridin-2-yl)propane-1,3-diamine (0.037 g, 0.19
mmol) and 4,5-dibromo-3-methylthiophene-2-carbaldehyde (0.054 g,
0.19 mmol) gave the title compound as a white solid (0.034 g, 49%).
m/z (ES+) 459 (100% M.sup.+).
Example 14
N-(4,5-Dibromo-3-methylthiophen-2-ylmethyl)-N'-(5H-imidazo[4,5-c]pyridazin-
-6-yl)propane-1,3-diamine dihydrochloride
[0104] a) [3-(5H-Imidazo[4,5-c]pyridazin-6-ylamino)propyl]carbamic
acid t-butyl ester. To a solution of 1,1'-thiocarbonyldiimidazole
(1.34 g, 7.5 mmol) in tetrahydrofuran (10 mL) at 0.degree. C. under
an atmosphere of argon was added dropwise, over a 15 minute period,
a solution of (3-aminopropyl)carbamic acid t-butyl ester (0.87 g, 5
mmol) in tetrahydrofuran (10 mL). After stirring at RT for 3 h the
mixture was evaporated and dissolved in dry dimethylformamide (10
mL) and treated with 3,4-diaminopyridazine. The resultant mixture
was heated under argon at 100.degree. C. After 48 h the mixture was
cooled and the DMF evaporated. Chromatography of the crude product
over silica gel eluting with dichloromethane containing increasing
concentrations of 10% ammonium hydroxide/methanol (0-12%) gave the
title compound as a cream coloured solid. m/z (ES+) 293 (51%,
MH.sup.+).
[0105] b) N-(5H-Imidazo[4,5-c]pyridazin-6-yl)propane-1,3-diamine
trifluoroacetate salt.
[3-(5H-Imidazo[4,5-c]pyridazin-6-ylamino)propyl]carbamic acid
t-butyl ester (0.27 g, 0.92 mmol) was treated with trifluoroacetic
acid (2 mL) at RT. After 2 h the mixture was evaporated, dissolved
in dichloromethane, re-evaporated and dried under vacuum to yield
pale yellow foam.
[0106] c)
N-(4,5-Dibromo-3-methylthiophen-2-ylmethyl)-N'-(5H-imidazo[4,5--
c]pyridazin-6-yl)propane-1,3-diamine dihydrochloride. Using the
general method for reductive amination
4,5-dibromo-3-methylthiophene-2-carbaldehyde (0.077 g, 0.27 mmol)
was reacted with
N.sup.1-(5H-imidazo[4,5-c]pyridazin-6-yl)propane-1,3-diamine
trifluoroacetate salt (0.082 g, 0.27 mmol) to give the title
compound, after conversion to the dihydrochloride with 1M
methanolic hydrogen chloride. 0.027 g. m/z (AP.sup.+) 459/461/463
(50/100/50%, MH.sup.+).
Example 15
N-(4,5-Dibromo-3-methylthiophen-2-ylmethyl)-N'-(1H-imidazo[4,5-b]pyridin-2-
-yl)propane-1,3-diamine
[0107] The 4,5-dibromo-3-methylthiophene-2-carbaldehyde was coupled
to compound 2a on a 0.2 mmol scale using the general method for
reductive amination to give the title compound as a white solid
(0.034 g, 49%). m/z (ES+) 459 (100% M.sup.+).
Example 16
N-(4-Bromo-3-methyl-5-vinylthiophen-2-ylmethyl)-N'-(5H-imidazo[4,5-c]pyrid-
azin-6-yl)-propane-1,3-diamine
[0108] a) 4-Bromo-3-methyl-5-vinylthiophene-2-carbaldehyde. A
mixture of 4,5-dibromo-3-methylthiophene-2-carbaldehyde (0.14 g,
0.5 mmol) tetrakis(triphenylphosphinyl)palladium (0.062 g, 0.05
mmol) and tri-n-butylvinylstannane (0.19 g, 0.6 mmol) in toluene
was heated at 100.degree. C. After 4 h the mixture was allowed to
cool and the solvent evaporated. The crude product was
chromatographed over silica gel eluting with hexane/dichloromethane
(95:5) to yield the title compound. m/z (CI.sup.+) 231/233
(MH.sup.+, 100:100%).
[0109] b)
N-(4-Bromo-3-methyl-5-vinylthiophen-2-ylmethyl)-N'-(5H-imidazo[-
4,5-c]pyridazin-6-yl)-propane-1,3-diamine. Using the general
procedure for reductive amination with compound 14b (0.030 g, 0.096
mmol) and 4-bromo-3-methyl-5-vinylthiophene-2-carbaldehyde (0.022
g, 0.096 mmol) gave the title compound as a white solid (0.010 g,
26%). m/z (ES.sup.+) 407 (100% M.sup.+).
Example 17
N-(4-Bromo-3-methyl-5-vinylthiophen-2-ylmethyl)-N'(1H-imidazo[4,5-b]pyridi-
n-2-yl)-propane-1,3-diamine
[0110] The 4-bromo-3-methyl-5-vinylthiophene-2, 16a, was coupled
with compound 2a on a 0.2 mmol scale using the general method for
reductive amination to give the title compound as a white solid
(0.010 g, 26%). m/z (ES.sup.+) 406 (100% M.sup.+).
Example 18
N-(4-Bromo-3-methyl-5-vinylthiophen-2-ylmethyl)-N'(1H-imidazo[4,5-c]pyridi-
n-2-yl)-propane-1,3-diamine
[0111] Aldehyde 16a was coupled with compound 13b on a 0.2 mmol
scale using the general method for reductive amination to give the
title compound as a white solid (0.010 g, 26%). m/z (ES.sup.+) 406
(100% M.sup.+).
Example 19
N-(3-Bromo-2-ethoxy-5-methylsulfanybenzyl)-N'-(5H-imidazo[4,5-c]pyridazin--
6-yl)propane-1,3-diamine dihydrochloride
[0112] a) 3-Bromo-2-ethoxy-5-iodobenzoic acid methyl ester. A
mixture of 3-bromo-2-hydroxy-5-iodobenzoic acid methyl ester (0.32
g, 0.9 mmol), potassium carbonate (0.38 g, 2.7 mmol) and iodoethane
in DMF was heated at 65.degree. C. After 48 h the mixture was
cooled, evaporated to dryness and the residue partitioned between
diethyl ether and water. The organic phase was separated, dried and
evaporated to yield the title compound. m/z (ES.sup.+)
385/387(MH.sup.+, 100:100%).
[0113] b) (3-Bromo-2-ethoxy-5-iodophenyl)methanol. To a solution of
3-bromo-2-ethoxy-5-iodobenzoic acid methyl ester (0.34 g, 0.88
mmol) in dry tetrahydrofuran (5 mL) at 0.degree. C. under argon was
added a 1M THF solution of diisobutylaluminium hydride (5.3 mL).
After 6 h the mixture was evaporated and partitioned between ethyl
acetate and 1M hydrochloric acid. The organic layer was separated,
dried and evaporated to yield the title compound. m/z (ES.sup.+)
379/381(MNa.sup.+, 100:100%).
[0114] c) 3-Bromo-2-ethoxy-5-iodobenzaldehyde. To a solution of
(3-bromo-2-ethoxy-5-iodophenyl)methanol (0.36 g, 0.78 mmol) in
dichloromethane was added maganese dioxide (0.068 g, 7.85 mmol)
After stirring at RT for 3 h the mixture was filtered and the
solvent evaporated. The crude product was chromatographed over
silica gel eluting with hexane/dichloromethane (1:1) to yield the
title compound. .delta..sub.H (CDCl.sub.3) 10.26 (1H, s); 8.10 (1H,
d); 8.07 (1H, d); 4.15 (2H, q); 1.49 (3H, t).
[0115] d) 2-(3-Bromo-2-ethoxy-5-iodophenyl)-1,3-dioxolane. A
solution of 3-bromo-2-ethoxy-5-iodobenzaldehyde (0.58 g, 1.63
mmol), 1,2-dihydroxyethane (0.11 g, 1.8 mmol) and 4-toluenesulfonic
acid monohydrate (0.01 g) in toluene (40 mL) was heated at reflux
with separation of water. After 14 h the mixture was cooled and the
solvent evaporated. The residue was partitioned between ethyl
acetate and aqueous sodium bicarbonate solution. The organic layer
was separated, dried and evaporated to yield the title compound.
m/z (ES.sup.+) 398/400 (M.sup.+, 100:100%).
[0116] e)
2-(3-Bromo-2-ethoxy-5-methylsulfanylphenyl)-1,3-dioxolane. A
mixture of 2-(3-bromo-2-ethoxy-5-iodophenyl)-1,3-dioxalane (0.43 g,
3.0 mmol), copper (I) oxide (0.095 g, 1.35 mmol) and sodium
thiomethoxide (0.09 g, 0.23 mmol) was heated at 80.degree. C. After
48 h the mixture was cooled, filtered and the solvent evaporated.
The residue was partitioned between water/diethyl ether the organic
layer separated, dried and evaporated to yield the title compound.
m/z (CI.sup.+) 319/321 (MH.sup.+, 100:100%).
[0117] f) 3-Bromo-2-ethoxy-5-methysulfanylbenzaldehyde. A solution
of 2-(3-bromo-2-ethoxy-5-methylsulfanylphenyl)-1,3-dioxalane (0.097
g, 0.3 mmol) and pyridinium toluenesulfonate (0.038 g, 0.15 mmol)
in acetone (10 mL) containing water (1 drop) was heated at reflux.
After 20 h the mixture was cooled, evaporated and the residue
partitioned between diethyl ether/aqueous sodium bicarbonate
solution. The organic phase was separated, dried and evaporated to
yield the title compound. .delta..sub.H (CDCl.sub.3) 10.30 (1H, s);
7.67 (1H, s); 7.44 (1H, d); 4.15 (2H, q); 2.50 (3H, s); 1.48 (3H,
t).
[0118] g)
N-(3-Bromo-2-ethoxy-5-methylsulfanybenzyl)-N'-(5H-imidazo[4,5-c-
]pyridazin-6-yl)propane-1,3-diamine dihydrochloride. Using the
general method for reductive amination
3-bromo-2-ethoxy-5-methysulfanylbenzaldehyde (0.05 g, 0.18 mmol)
was reacted with compound 14b (0.055 g, 0.182 mmol) to give the
title compound, after conversion to the dihydrochloride with 1M
methanolic hydrogen chloride. 0.035 g. m/z (AP.sup.+) 451/453
(100/100% MH.sup.+).
Example 20
N-(6-Chloro-8-iodochroman-4-yl)-N'-(5H-imidazo[4,5-c]pyridazin-6-yl)propan-
e-1,3-diamine
[0119] Using the method described in Example 3
6-chloro-8-iodochromanone (0.05 g, 0.27 mmol) was reacted with
compound 14b (0.035 g, 0.11 mmol) to give the title compound. 0.007
g, m/z (AP.sup.+) 480/482(100/35%, MH.sup.+).
Example 21
N-(3-Chloro-5-methoxy-1H-indol-7-ylmethyl)-N'-(5H-imidazo[4,5-c]pyridazin--
6-yl)propane-1,3-diamine dihydrochloride
[0120] Using the general method for reductive amination, aldehyde
12a (0.018 g, 0.09 mmol) was reacted with compound 14b (0.028 g,
0.09 mmol) to give the title compound, after conversion to the
dihydrochloride with 1M methanolic hydrogen chloride. 0.015 g, m/z
(AP.sup.+) 386/388 (100/35%, MH.sup.+).
Example 22
N-(4,5-Dibromo-3-methylthiophen-2-ylmethyl)-N'-(1H-thieno[3,4-d]imidazol-2-
-yl)propane-1,3-diamine
[0121] a) 3,4-Diaminothiophene. To a suspension of
2,5-dibromo-3,4-dinitrothiophene (10 g, 30.12 mmol) in conc. HCl
(294 mL) was added portionwise tin (21.2 g, 178 mmol) keeping the
temperature below 30.degree. C. The mixture was stirred for 4 h at
room temperature then stored in the fridge for two days. The solid
was collected by filtration, washed with diethyl ether and acetone
then suspended in water (60 mL) and diethylether (60 mL). The
mixture was cooled in an ice-bath, then made alkaline using 4M
NaOH. The aqueous phase was separated then continuously extracted
with diethyl ether. The organic phases were combined then
concentrated to give 2.3 g of the title compound as a beige solid;
.delta..sub.H (CDCl.sub.3) 6.1 (2H, s), 3.3 (4H, bs).
[0122] b) {3-[3-(4-Aminothiophen-3-yl)thioureido]propyl}carbamic
acid tert-butyl ester. The product from 22a (0.421 g, 3.7 mmol) was
dissolved in dichloromethane (13 mL) and treated with
diisopropylethylamine (892 uL, 5.14 mmol), 4-dimethylaminopyridine
(0.125 g, 1 mmol) and (3-isothiocyanato-propyl)carbamic acid
tert-butyl ester (1.14 g, 5.28 mmol). The reaction mixture was
stirred at room temperature for 16 h, loaded on Kieselgel 60 and
chromatographed eluting with 0-2% (9:1 MeOH/20 M NH.sub.3) in
dichloromethane. The fractions containing the title compound were
combined and concentrated, the residue was re-chromatographed on
Kieselgel 60 eluting with 0-50% ethyl acetate in hexane. The
fractions containing the title compound were combined and
concentrated, the residue was applied to a 10 g Varian Bond Elute
SCX cartridge which was flushed with methanol. The cartridge was
then eluted with 0.2 M NH.sub.3 in methanol, and this eluate
evaporated to dryness to afford the title compound as a pale yellow
oil (0.26 g, 21%), m/z (CI.sup.+) 331 (MH.sup.+, 10%).
[0123] c) [3-(1H-Thieno[3,4-d]imidazol-2-ylamino)propyl]carbamic
acid tert-butyl ester. The product from 22b (0.180 g, 0.54 mmol)
was dissolved in dimethylformamide (40 mL) and treated with
triethylamine (76 uL, 0.54 mmol) and mercuric chloride (0.149 g,
0.54 mmol). The reaction mixture was stirred at room temperature
for 16 h then diluted with ethyl acetate and filtered through
celite. The filtrate was concentrated and the residue
chromatographed on Kieselgel 60 eluting with 0-10% (9:1 MeOH/20 M
NH.sub.3) in CH.sub.2Cl.sub.2 to afford the title compound as a
brown oil (0.050 g, 31%), m/z (CI.sup.+) 297 (MH.sup.+, 100%).
[0124] d) N-(1H-Thieno[3,4-d]imidazol-2-yl)propane-1,3-diamine
bistrifluoroacetate salt. The product from 22c (0.045 g, 0.15 mmol)
was dissolved in trifluoroacetic acid (2 mL). The reaction mixture
was stirred at room temperature for 0.5 h then concentrated to
afford the title compound as a brown oil (0.056 g, 88%), m/z
(CI.sup.+) 197 (MH.sup.+, 80%), 393 (2 MH.sup.+, 100%).
[0125] e)
N-(4,5-Dibromo-3-methylthiophen-2-ylmethyl)-N'-(1H-thieno[3,4-d-
]imidazol-2-yl)propane-1,3-diamine. The product from 22d was
coupled to 4,5-dibromo-3-methylthiophene-2-carbaldehyde on a 0.09
mmol scale using the general method for reductive amination to give
the title compound as a pink solid (0.015 g, 36%), m/z (CI.sup.+)
465 (MH.sup.+, 100%).
Example 23
N-(4-Bromo-3-methyl-5-vinylthiophen-2-ylmethyl)-N'-(1H-thieno[3,4-d]imidaz-
ol-2-yl)propane-1,3-diamine
[0126] The product from 22d was coupled to aldehyde 16a on a 0.09
mmol scale using the general method for reductive amination to give
the title compound as a pink solid (0.015 g, 36%), m/z (CI.sup.+)
411 (MH.sup.+, 100%).
Example 24
N-(3-Chloro-5-methoxy-1H-indol-7-ylmethyl)-N'-(1H-thieno[3,4-d]imidazol-2--
yl)propane-1,3-diamine
[0127] The product from 19d was coupled to compound 12a on a 0.13
mmol scale using the general method for reductive amination to give
the title compound as a beige solid (0.038 g, 73%), m/z (CI.sup.+)
390 (MH.sup.+, 70%).
Example 25
N-(4-Bromo-5-difluoromethyl-3-methylthiophen-2-ylmethyl)-N'-(1H-imidazo[4,-
5-b]pyridin-2-yl)propane-1,3-diamine dihydrochloride
[0128] a) 2-(4,5-Dibromo-3-methylthiophen-2-yl)-1,3-dioxolane. A
solution of 4,5-dibromo-3-methylthiophene-2-carbaldehyde (2.84 g,
10 mmol), 1,2-dihydroxyethane (1.36 g, 22 mmol) and resin supported
4-toluenesulfonic acid in toluene (100 mL) was heated at reflux
with separation of water. After 3 h the mixture was cooled,
filtered and the solvent evaporated to yield the title compound
(3.28 g); m/z (AP.sup.+) 329 (MH.sup.+, 100%).
[0129] b)
3-Bromo-5-(1,3-dioxolan-2-yl)-4-methylthiophene-2-carbaldehyde.
Compound 25a (3.28 g, 10 mmol) was dissolved in dry THF (40 mL) and
cooled to -78.degree. C. under an argon atmosphere. A solution of
n-butyl lithium (2.5 M in cyclohexane, 4 mL) was added dropwise.
After stirring at -78.degree. C. for 0.3 h, the solution was
treated with dry DMF (0.775 mL, 10 mmol) and stirred for a further
0.6 h. The cooling bath was then removed and the solution allowed
to reach room temperature over 3 h. The reaction mixture was
quenched with 2N aqueous HCl and the product extracted into
dichloromethane. The extracts were combined, dried (MgSO.sub.4) and
evaporated to yield the title compound (2.4 g); .delta..sub.H
(CDCl.sub.3) 2.29 (3H, s, CH.sub.3), 4.07 (4H, m,
CH.sub.2CH.sub.2), 6.14 (1H, s, CH), 9.98 (1H, s, CHO).
[0130] c)
2-(4-Bromo-5-difluoromethyl-3-methylthiophen-2-yl)-1,3-dioxolan- e.
A solution of compound 25b (0.277 g, 1 mmol) and deoxofluor (0.378
g, 1.7 mmol) in dichloromethane (1 ml) was heated at reflux. After
14 h the mixture was diluted with dichloromethane, washed
successively with aqueous sodium bicarbonate and 1N aqueous HCl,
dried (MgSO.sub.4) and evaporated to yield the title compound (0.28
g); .delta..sub.H (CDCl.sub.3) 2.25 (3H, s, CH.sub.3), 4.06 (4H, m,
CH.sub.2CH.sub.2), 6.12 (1H, s, CH), 6.87 (1H, t, J=54.8 Hz,
CHF.sub.2).
[0131] d)
4-Bromo-5-difluoromethyl-3-methylthiophene-2-carbaldehyde. A
solution of compound 25c (0.598 g, 2 mmol) and resin supported
4-toluenesulfonic acid in acetone (20 mL) containing water (1 mL)
was stirred at room temperature. After 14 h the mixture was
filtered and the solvent evaporated to yield the title compound
(0.365 g); .delta..sub.H (CDCl.sub.3) 2.56 (3H, s, CH.sub.3), 6.90
(1H, t, J=56 Hz, CHF.sub.2), 10.07 (1H, s, CHO).
[0132] e)
N-(4-Bromo-5-difluoromethyl-3-methylthiophen-2-ylmethyl)-N'-(1H-
-imidazo[4,5-b]pyridin-2-yl)propane-1,3-diamine dihydrochloride.
Using the general method for reductive amination, compound 25d
(0.074 g, 0.29 mmol) was reacted with compound 2a (0.056 g, 0.29
mmol) to give the title compound, after conversion to the
dihydrochloride with 1M methanolic hydrogen chloride (0.043 g); m/z
(AP.sup.+) 430 (MH.sup.+, 100%).
Example 26
N-(4-Bromo-5-difluoromethyl-3-methylthiophen-2-ylmethyl)-N'-(5H-imidazo[4,-
5-c]pyridazin-6-yl)propane-1,3-diamine dihydrochloride
[0133] Using the general method for reductive amination, compound
25d (0.074 g, 0.29 mmol) was reacted with compound 14b (0.29 mmol)
to give the title compound, after conversion to the dihydrochloride
with 1M methanolic hydrogen chloride (0.037 g); m/z (AP.sup.+) 431
(MH.sup.+, 100%).
Example 27
N-(4-Bromo-5-difluoromethyl-3-methylthiophen-2-ylmethyl)-N'-(1H-thieno[3,4-
-d]imidazol-2-yl)propane-1,3-diamine dihydrochloride
[0134] Using the general method for reductive amination, compound
25d (0.061 g, 0.24 mmol) was reacted with compound 19d (0.24 mmol)
to give the title compound, after conversion to the dihydrochloride
with 1M methanolic hydrogen chloride (0.044 g); m/z (AP.sup.+) 435
(MH.sup.+, 100%).
Example 28
N-(4-Bromo-3-methyl-5-trifluoromethylthiophen-2-ylmethyl)-N'-(5H-imidazo[4-
,5-c]pyridazin-6-yl)propane-1,3-diamine dihydrochloride
[0135] a)
4-Bromo-3-methyl-5-trifluoromethylthiophene-2-carbaldehyde.
4,5-Dibromo-3-methylthiophene-2-carbaldehyde (250 mg, 0.88 mmol)
was dissolved in a mixture of DMF (10 mL) and N-methylpyrrolidine
(0.5 mL) containing copper (I) iodide (200 mg, 0.96 mmol) and
2,2-difluoro-2-fluorosulfonyl acetic acid methyl ester (871 mg, 4.4
mmol). After heating at 70.degree. C. with vigorous stirring for 7
h the mixture was cooled and allowed to cool to room temperature
overnight. The DMF was evaporated and the residue was partitioned
between diethyl ether (30 mL) and saturated ammonium chloride
solution. The organic layer was separated, washed with brine, dried
and evaporated to yield the crude product. Filtration through
silica gel eluting with hexane/diethyl ether (1:1) gave the title
compound as a pale yellow solid (155 mg); m/z (AP.sup.-) 272
(M.sup.-, 25%).
[0136] b)
N-(4-Bromo-3-methyl-5-trifluoromethylthiophen-2-ylmethyl)-N'-(5-
H-imidazo[4,5-c]pyridazin-6-yl)propane-1,3-diamine dihydrochloride.
Using the general method for reductive amination, purification and
salt formation, compound 28a was reacted with compound 14b to give
the title compound (23 mg); m/z (AP.sup.+) 449 (MH.sup.+,
100%).
Example 29
N-(4-Bromo-3-methyl-5-trifluoromethylthiophen-2-ylmethyl)-N'-(1H-thieno[3,-
4-d]imidazol-2-yl)propane-1,3-diamine dihydrochloride
[0137] Using the general method for reductive amination, compound
28a (0.066 g, 0.24 mmol) was reacted with compound 19d (0.24 mmol)
to give the title compound, after conversion to the dihydrochloride
with 1M methanolic hydrogen chloride (0.010 g); m/z (AP.sup.+) 453
(MH.sup.+, 100%).
Example 30
N-(4-Bromo-3-methyl-5-trifluoromethylthiophen-2-ylmethyl)-N'-(1H-imidazo[4-
,5-b]pyridin-2-yl)propane-1,3-diamine dihydrochloride
[0138] Using the general method for reductive amination, compound
28a (0.066 g, 0.24 mmol) was reacted with compound 2a (0.24 mmol)
to give the title compound, after conversion to the dihydrochloride
with 1M methanolic hydrogen chloride (0.012 g); m/z (AP.sup.+) 448
(MH.sup.+, 100%).
Example 31
N-(4-Bromo-3-methyl-5-trifluoromethylthiophen-2-ylmethyl)-N'-(1H-imidazo[4-
,5-c]pyridin-2-yl)propane-1,3-diamine dihydrochloride
[0139] Using the general method for reductive amination, compound
28a (0.066 g, 0.24 mmol) was reacted with compound 13a (0.24 mmol)
to give the title compound, after conversion to the dihydrochloride
with 1M methanolic hydrogen chloride (0.010 g); m/z (AP.sup.+) 448
(MH.sup.+, 100%).
Example 32
N-(4-Bromo-5-(1-fluorovinyl)-3-methylthiophen-2-ylmethyl)-N'-(1H-imidazo[4-
,5-b]pyridin-2-yl)propane-1,3-diamine hydrochloride
[0140] a) 4-Bromo-3-methyl-5-vinyl-thiophene-2-carbaldehyde. A
solution of 4,5-dibromo-3-methylthiophene-2-carbaldehyde (1.42 g, 5
mmol), tetrakis(triphenylphosphine)-palladium(0) (0.622 g, 0.5
mmol) and tributylvinyltin (1.75 mL, 6 mmol) in toluene (50 mL) was
heated at 100.degree. C. After 14 h the reaction mixture was
concentrated and the residue was chromatographed on Kieselgel 60
eluting with 0-50% dichloromethane in hexane to afford the title
compound (0.623 g); m/z (AP.sup.+) 231 (MH.sup.+, 100%).
[0141] b)
4-Bromo-5-(2-bromo-1-hydroxyethyl)-3-methylthiophene-2-carbalde-
hyde. A solution of compound 32a (0.42 g, 1.82 mmol) in
dimethylsulfoxide (2.1 mL) was treated successively with water
(0.067 mL, 3.64 mmol) and N-bromosuccinimide (0.647 g, 3.64 mmol).
After 0.5 h, aqueous sodium bicarbonate was added and the product
extracted with ethyl acetate. The extracts were combined, dried
(MgSO.sub.4) and evaporated. The residue was chromatographed on
Kieselgel 60 eluting with 0-4% ethyl acetate in dichloromethane to
afford the title compound (0.375 g); .delta..sub.H (CDCl.sub.3)
2.54 (3H, s, CH.sub.3), 2.97 (1H, d, J=3.8 Hz, OH), 3.54 (1H, dd,
J=8.6, 10.6 Hz, CH.sub.2Br), 3.82 (1H, dd, J=3.1, 8.6 Hz,
CH.sub.2Br), 5.27 (1H, m, CHOH), 10.07 (1H, s, CHO).
[0142] c)
4-Bromo-5-(1-fluorovinyl)-3-methylthiophene-2-carbaldehyde.
[0143] Method A. Diphenyl(1-fluorovinyl)methylsilane. In a
flame-dried 1 L 3-neck round bottom under an anhydrous atmosphere,
65 mL (309 mmol) of diphenylmethylchlorosilane was added to 4.3 g
(618 mmol) of lithium wire in 650 mL of anhydrous THF. The mixture
was stirred at ambient temperature for 20 hours. The mixture was
then cooled to -78.degree. C. and the atmosphere replaced with
1,1-difluoroethylene (excess) such that the temperature of the
reaction mixture remained below -55.degree. C. Difluoroethylene
addition was stopped when the reaction temperature remained at or
below -70.degree. C. The reaction was stirred at <-70.degree. C.
until it turned a clear light yellow (.about.2 hr.) and was then
allowed to warm to ambient temperature. The remaining lithium wire
was removed and the mixture treated with portions of
Na.sub.2SO.sub.4-10H.sub.2O until no gas evolved upon addition. The
mixture was then dried over Na.sub.2SO.sub.4, filtered through a
silica pad and the pad rinsed with ether. The combined filtrates
were dried under vacuum, the resulting residue suspended/dissolved
in hexanes and filtered through another silica pad. The pad was
rinsed with hexanes, the filtrates combined and the solvent removed
under reduced pressure to give a light yellowish liquid with some
white crystalline material present. The product was purified by
vacuum distillation (113-117.degree. C. at .about.2 Torr) to give
44 g (59%) of the title compound as a clear colorless liquid.
.delta..sub.H (CDCl.sub.3): 0.72 (3H, s, CH.sub.3), 4.85 (1H, dd,
J=2.6, 61.2 Hz, CH.sub.2), 5.48 (1H, dd, J=2.6, 33.3, CH.sub.2),
7.39 (6H, m, ArH), 7.59 (4H, d, J=6.8 Hz, ArH); .delta..sub.F
(CDCl.sub.3): -103.16 (q, dd, J=33.3, 61.2 Hz).
[0144] b)
4-Bromo-5-(1-fluorovinyl)-3-methylthiophene-2-carbaldehyde. Under
an inert atmosphere in a 25 mL round bottom flask were combined 166
mg of Diphenyl(1-fluorovinyl)methylsilane (0.685 mmol), 130 mg of
4,5-dibromo-3-methylthiophene-2-carbaldehyde (0.459 mmol), 209 mg
of CsF (1.38 mmol), 88 mg of CuI (0.459 mmol), 10.5 mg
Pd.sub.2(dba).sub.3 (0.0115 mmol) and 14.1 mg AsPh.sub.3 (0.0459
mmol). The flask containing the solids was cooled to
.about.0.degree. C. with an ice bath and 2 mL of degassed,
anhydrous dimethylformamide (DMF) were added. The reaction mixture
was stirred at 0 to 5.degree. C. for 2 hr and then 2 mL of water
was added. The mixture was then diluted with 5 mL 1N NaOH and
extracted with 25% diethyl ether/hexanes (4.times.20 mL). The
combined extracts were washed with brine (1.times.5 mL), dried over
Na.sub.2SO.sub.4, and the solvent removed under vacuum. The
remaining residue was purified by flash silica gel chromatography
(CH.sub.2Cl.sub.2/hexanes) to give a 50% yield of the title
compound as a white solid. .delta..sub.H (CDCl.sub.3) 2.57 (3H, s,
CH.sub.3), 5.24 (1H, dd, J=4.0, 18.5 Hz, CH.sub.2), 5.70 (1H, dd,
J=4.0, 49.6 Hz, CH.sub.2), 10.06 (1H, s, CHO); .delta..sub.F
(CDCl.sub.3): -92.20 (q, dd, J=18.4, 50.4 Hz); m/z (ESI.sup.+)
(MH.sup.+, 249).
[0145] Method B. A solution of compound 32b (0.384 g, 1.16 mmol) in
dry dichloromethane (6 mL) was cooled to -78.degree. C. A solution
of deoxofluor (0.282 g, 1.27 mmol) in dry dichloromethane (4 mL)
was added. After 3 h the mixture was diluted with dichloromethane,
washed with aqueous sodium bicarbonate, dried (MgSO.sub.4) and
evaporated. The residue was dissolved in benzene (15 mL) and
treated with DBU (0.1 g, 0.66 mmol). After 2 h the reaction mixture
was concentrated, the residue was diluted with dichloromethane and
washed with 1M aqueous HCl. The organic phase was dried
(MgSO.sub.4) and concentrated to yield the title compound (0.133
g); .delta..sub.H (CDCl.sub.3) 2.57 (3H, s, CH.sub.3), 5.23 (1H,
dd, J=4.0, 18.5 Hz, CH.sub.2), 5.70 (1H, dd, J=4.0, 49.9 Hz,
CH.sub.2), 10.04 (1H, s, CHO).
[0146] d)
N-(4-Bromo-5-(1-fluorovinyl)-3-methylthiophen-2-ylmethyl)-N'-(1-
H-imidazo[4,5-b]pyridin-2-yl)propane-1,3-diamine hydrochloride.
Using the general method for reductive amination, compound 32c
(0.075 g, 0.3 mmol) was reacted with compound 2a (0.057 g, 0.3
mmol) to give the title compound, after conversion to the
hydrochloride with 1M methanolic hydrogen chloride (0.026 g); m/z
(AP.sup.+) 424 (MH.sup.+, 100%).
Example 33
N-(4-Bromo-5-(1-fluorovinyl)-3-methylthiophen-2-ylmethyl)-N'-(1H-imidazo[4-
,5-c]pyridin-2-yl)propane-1,3-diamine
[0147] Using the general method for reductive amination, compound
32c (0.075 g, 0.3 mmol) was reacted with compound 13a (0.057 g, 0.3
mmol) to give the title compound (0.025 g); m/z (AP.sup.+) 424
(MH.sup.+, 100%).
Example 34
N-(4-Bromo-5-(1-fluorovinyl)-3-methylthiophen-2-ylmethyl)-N'-(5H-imidazo[4-
,5-c]pyridazin-6-yl)propane-1,3-diamine
[0148] Using the general method for reductive amination, compound
32c (0.066 g, 0.27 mmol) was reacted with compound 14b (0.27 mmol)
to give the title compound (0.022 g); m/z (AP.sup.+) 425 (MH.sup.+,
100%).
Example 35
N-(4-Bromo-5-(1-fluorovinyl)-3-methylthiophen-2-ylmethyl)-N'-(1H-thieno[3,-
4-d]imidazol-2-yl)propane-1,3-diamine
[0149] Using the general method for reductive amination, compound
32c (0.059 g, 0.24 mmol) was reacted with compound 19d (0.24 mmol)
to give the title compound (0.025 g); m/z (AP.sup.+) 429 (MH.sup.+,
100%).
Example 36
N-(4-Bromo-3-methyl-5-ethynylthiophen-2-ylmethyl)-N'-(5H-imidazo[4,5-c]pyr-
idazin-6-yl)propane-1,3-diamine
[0150] a) 4-Bromo-5-ethynyl-3-methylthiophene-2-carbaldehyde. A
mixture of trimethylsilylacetylene (3.7 mL, 26.4 mmol),
4,5-dibromo-3-methylthiophene-2-carbaldehyde (1.50 g, 52.8 mmol),
copper (I) iodide (19 mg) and bis(triphenylphosphine)palladium (II)
chloride (80 mg) in triethylamine (35 mL) was stirred at room
temperature. After 2 h the mixture was filtered and evaporated. The
crude product was chromatographed over silica gel eluting with
petroleum ether 40-60 containing increasing concentrations of
dichloromethane up to 50%. The resultant trimethylsilyl protected
acetylene was treated with methanol containing 10% concentrated
ammonium hydroxide. After stirring at 20.degree. C. for 15 minutes
the solvent was evaporated to yield the title compound as a light
brown foam (0.73 g); .delta..sub.H (CDCl.sub.3) 9.99 (1H, s), 3.80
(1H, s), 2.55 (3H, s).
[0151] b)
N-(4-Bromo-3-methyl-5-ethynylthiophen-2-ylmethyl)-N'-(5H-imidaz-
o[4,5-c]pyridazin-6-yl)propane-1,3-diamine. Using the general
method for reductive amination and purification, compound 36a was
reacted with compound 14b (0.24 mmol) to give the title compound
(21 mg); m/z (AP.sup.+) 405 (MH.sup.+, 100%).
Example 37
N-(4-Bromo-3-methyl-5-ethynylthiophen-2-ylmethyl)-N'-(1H-imidazo[4,5-b]pyr-
idin-2-yl)propane-1,3-diamine
[0152] Using the general method for reductive amination and
purification, compound 36a was reacted with compound 2a (0.24 mmol)
to give the title compound (19 mg); m/z (AP.sup.+) 404 (MH.sup.+,
100%).
Example 38
N-(4-Bromo-3-methyl-5-ethynylthiophen-2-ylmethyl)-N'-(1H-imidazo[4,5-c]pyr-
idin-2-yl)propane-1,3-diamine
[0153] Using the general method for reductive amination and
purification, compound 36a was reacted with compound 13a (0.24
mmol) to give the title compound (23 mg); m/z (AP.sup.+) 404
(MH.sup.+, 100%).
Example 39
N-(4-Bromo-3-methyl-5-ethynylthiophen-2-ylmethyl)-N'-(1H-thieno[3,4-d]imid-
azol-2-yl)propane-1,3-diamine
[0154] Using the general method for reductive amination and
purification, compound 36a was reacted with compound 19d (0.24
mmol) to give the title compound (23 mg); m/z (AP.sup.+) 409
(MH.sup.+, 100%).
Example 40
N-(4-Bromo-5-ethyl-3-(1-propynyl)thiophen-2-ylmethyl)-N'-(5H-imidazo[4,5-c-
]pyridazin-6-yl)propane-1,3-diamine dihydrochloride
[0155] a) 3,4-Dibromo-5-ethylthiophene-2-carbaldehyde.
2,3,4-Tribromo-5-ethylthiophene (1.4 g, 4.1 mmol) was dissolved in
dry THF (40 mL) and cooled to -78.degree. C. under an argon
atmosphere. A solution of n-butyl lithium (1.6 M in cyclohexane,
2.56 mL) was added dropwise. After stirring at -78.degree. C. for
0.5 h, the solution was treated with dry DMF (0.32 mL, 4.1 mmol)
and stirred for a further 0.6 h. The cooling bath was then removed
and the solution allowed to reach room temperature over 3 h. The
reaction mixture was quenched with 2M aqueous HCl and the product
extracted into dichloromethane. The extracts were combined, dried
(MgSO.sub.4) and evaporated. The residue was chromatographed on
Kieselgel 60 eluting with 0-50% dichloromethane in hexane to yield
the title compound (0.6 g); m/z (AP.sup.+) 299 (MH.sup.+,
100%).
[0156] b) 4-Bromo-3-(1-propynyl)-5-ethylthiophene-2-carbaldehyde.
Methyl acetylene was bubbled into a solution containing
triethylamine (2 ml) and THF (2 ml). After 10 min the solution was
quickly added to a 10 mL reactor vessel containing compound 40a
(0.227 g, 0.8 mmol), bis(triphenylphosphine)palladium (II) chloride
(5.8 mg, 0.008 mmol) and copper (I) iodide (0.15 mg, 0.0008 mmol).
The reactor was sealed and the reaction mixture stirred at room
temperature. After 60 h, the reaction mixture was filtered through
celite and concentrated. The residue was chromatographed on
Kieselgel 60 eluting with 0-50% dichloromethane in hexane to yield
the title compound (0.126 g); m/z (AP.sup.+) 257 (MH.sup.+,
100%).
[0157] c)
N-(4-Bromo-5-ethyl-3-(1-propynyl)thiophen-2-ylmethyl)-N'-(5H-im-
idazo[4,5-c]pyridazin-6-yl)propane-1,3-diamine dihydrochloride.
Using the general method for reductive amination, compound 40b
(0.051 g, 0.2 mmol) was reacted with compound 14b (0.2 mmol) to
give the title compound, after conversion to the dihydrochloride
with 1M methanolic hydrogen chloride (0.024 g); m/z (AP.sup.+)
433/435 (MH.sup.+, 100%).
Example 41
N-(4-Bromo-5-ethyl-3-(1-propynyl)thiophen-2-ylmethyl)-N'-(1H-imidazo[4,5-c-
]pyridin-2-yl)propane-1,3-diamine
[0158] Using the general method for the synthesis of example
40.
Example 42
N-(4-Bromo-5-ethyl-3-(1-propynyl)thiophen-2-ylmethyl)-N'-(1H-imidazo[4,5-b-
]pyridin-2-yl)propane-1,3-diamine
[0159] Using the general method for the synthesis of example
40.
Example 43
N-(4-Bromo-5-ethyl-3-(1-propynyl)thiophen-2-ylmethyl)-N'-(1H-thieno[3,4-d]-
imidazol-2-yl)propane-1,3-diamine
[0160] Using the general method for the synthesis of example
40.
Biological Data
1. Enzyme Inhibition (S. aureus MRS)--Aminoacylation Assay
[0161] Compounds of the present invention may be assayed for their
ability to inhibit the enzyme methionyl tRNA synthetase (MRS),
using recombinant S. aureus MRS, as follows: TABLE-US-00002
Reaction Mix (per 1 ml) Stock Volume (.mu.l) Final Concentration
100 mM Tris/Cl, pH 7.9 600 30 mM 250 mM KCl 75 mM 125 mM ATP 40 2.5
mM 250 mM MgCl.sub.2 80 10 mM 50 mM DTT 80 2 mM 1 mM Met (H-3 hot
and cold) 40 10 uM Solid tRNA 4 mg/ml 2 mg/ml (Mixed E. coli MRE
600) H.sub.2O 160 10.times. Inhibitor (0-10 .mu.M) 5 .mu.l per well
0-1 .mu.M
[0162] The reaction is started by adding 20 .mu.l appropriately
diluted pure enzyme (pre-incubated with inhibitor) to 25 .mu.l
reaction mix for 10 min at room temperature. The reaction is
terminated by the addition of 150 .mu.l 167 mM sodium citrate, pH
2.15 containing phosphodiesterase (PDE) SPA beads (0.833 mg/ml).
The binding of the radiolabelled product to the bead brings the
isotope into close enough proximity to allow radiation from the
tritium to excite the scintillant within the bead. Any unbound
radiolabel is not close enough to the scintillant to allow this
energy transfer, so no signal is generated. Following termination
of the reaction, plates are spun at 2500 rpm for 5 min in a Mistral
3000E plate centrifuge (or alternatively allowed to stand for 1
hour). The assay is conducted in 96-well Optiplates (Packard).
Plates are counted on a TopCount. (Packard 96 well counter).
Reagents
[0163] Mixed E. coli MRE 600 tRNA and ATP were purchased from
Boehringer-Mannheim, L-[methyl-.sup.3H]methionine and
phosphodiesterase scintillation proximity (SPA) beads from Amersham
Pharmacia Biotech and other reagents from Sigma.
[0164] Pure recombinant S. aureus MRS (EP application number
97300317.1, SmithKline Beecham) was obtained using standard
purification procedures. The enzyme is diluted in Dilution Buffer
which consists of 10 mM Tris/Cl, 2 mM DTT pH 7.9.
Results
[0165] Examples 1 to 7, 9, 11, 12, 15, 17, 22-30, 32, 34-37, 39-41,
and 43-44 have IC.sub.50 values against S. aureus MRS in the range
<3 to 200 nM. All are highly selective with respect to the
mammalian enzyme (no inhibition of rat MRS up to 1 .mu.M).
2. Enzyme Inhibition (H. influenzae MRS)--Aminoacylation Assay
[0166] Compounds of the present invention may be assayed for their
ability to inhibit the enzyme methionyl tRNA synthetase (MRS),
using recombinant H. influenzae MRS (R. D. Fleischmann et al.,
Science, 269, 496-512, 1995), as follows: TABLE-US-00003 Reaction
Mix (per 1 ml) Stock Volume (.mu.l) Final Concentration 100 mM
Tris/Cl, pH 7.9 600 30 mM 250 mM KCl 75 mM 125 mM ATP 40 2.5 mM 250
mM MgCl.sub.2 80 10 mM 50 mM DTT 80 2 mM 1 mM Met (H-3 hot and
cold) 20 10 .mu.M Solid tRNA 4 mg/ml 2 mg/ml (Mixed E. coli MRE
600) H.sub.2O 180 10.times. Inhibitor (0-100 .mu.M) 5 .mu.l per
well 0-10 .mu.M
[0167] The reaction is started by adding 20 .mu.l appropriately
diluted pure enzyme (pre-incubated with inhibitor) to 25 .mu.l
reaction mix for 10 min at room temperature. The reaction is
terminated by the addition of 150 .mu.l 167 mM sodium citrate, pH
2.15 containing phosphodiesterase (PDE) SPA beads (0.833 mg/ml).
The binding of the radiolabelled product to the bead brings the
isotope into close enough proximity to allow radiation from the
tritium to excite the scintillant within the bead. Any unbound
radiolabel is not close enough to the scintillant to allow this
energy transfer, so no signal is generated. Following termination
of the reaction, plates are spun at 2500 rpm for 5 min in a Mistral
3000E plate centrifuge (or alternatively allowed to stand for 1
hour). The assay is conducted in 96-well Optiplates (Packard).
Plates are counted on a TopCount. (Packard 96 well counter).
Reagents
[0168] Mixed E. coli MRE 600 tRNA and ATP were purchased from
Boehringer-Mannheim, L-[methyl-.sup.3H]methionine and
phosphodiesterase scintillation proximity (SPA) beads from Amersham
Pharmacia Biotech and other reagents from Sigma.
Results
[0169] Examples 5, 8, 13-4, and 28-44, have IC.sub.50 values
against H. influenzae MRS in the range <3 to 2200 nM. All are
highly selective with respect to the mammalian enzyme (no
inhibition of rat MRS up to 1 .mu.M).
3. Antibacterial Activity
[0170] Compounds of the present invention were assayed for
antibacterial activity against a range of pathogenic organisms
(strains of S. aureus, S. pneumoniae, E. faecalis, H. influenzae
and M. catarrhalis) in a standard MIC assay modified by the
inclusion of cyclodextrin, to assist with solubility.
[0171] Examples 1-4, 6, 9, 11, 12, 15, 17, 22-30, 32, 34-37, 39-41,
and 43-44 had MIC's <1 .mu.g/ml against some strains of the
organisms S. aureus, S. pneumoniae, and E. faecalis. Examples 2,
13-24, and 28-44 had MIC's<8 .mu.g/ml against some strains of
the organisms M. catarrhalis and H. influenzae.
[0172] Compound 3 was tested against a wider range of clinical
isolates of S. aureus, Staphylococcus epidermidis, E. faecalis and
Enterococcus faecium to determine MIC90 values (the concentration
required to inhibit 90% of the organisms). The panels of isolates
included a large proportion of organisms resistant to various
clinical antibiotics. (See Jarvest, et al., J. Med. Chem., 2002,
45, 1959). Very good activity was seen against all the organisms,
with all MIC90 values at .ltoreq.1 .mu.g/ml (MIC90's: S. aureus, 1
.mu.g/ml; S. epidermidis, 0.5 .mu.g/ml; E. faecalis, 0.06 .mu.g/ml;
and E. faecium 0.03 .mu.g/ml).
[0173] The enantiomers of 3 (11(g) and 11(h)) were assayed in the
usual way. The (R)-enantiomer was found to be the more active
isomer with a lower IC.sub.50 value and potent antibacterial
activity. The compression of IC.sub.50 values <10 nM due to the
limit of the enzyme concentration in the assay (3 nM) (see Jarvest,
et al., J. Med. Chem., 2002, 45, 1959) makes it hard to calculate
the enantiomeric inhibitory ratio. However, the ratio of the
antibacterial activity of the two isomers suggests a high degree of
enantioselectivity, of the order of at least two orders of
magnitude.
[0174] In conclusion, the key right hand side pharmacophore for
bacterial MRS inhibition has been defined as an NH--C--NH unit in
the context of a bicyclic heteroaromatic system. Potent
non-quinolone analogues have been obtained with excellent
antibacterial activity against staphylococci and enterococci,
including antibiotic resistant isolates. In addition, the
biologically active configuration of the tetrahydroquinoline series
has been identified as possessing (R) stereochemistry.
* * * * *