U.S. patent application number 09/725391 was filed with the patent office on 2002-04-25 for farnesyl transferase inhibiting 1,8-annelated quinolinone derivatives substituted with n- or c-linked imidazoles.
Invention is credited to Angibaud, Patrick Rene, Ligny, Yannick Aime Eddy, Poncelet, Virginie Sophie, Sanz, Gerard Charles, Venet, Marc Gaston.
Application Number | 20020049327 09/725391 |
Document ID | / |
Family ID | 26146222 |
Filed Date | 2002-04-25 |
United States Patent
Application |
20020049327 |
Kind Code |
A1 |
Venet, Marc Gaston ; et
al. |
April 25, 2002 |
Farnesyl transferase inhibiting 1,8-annelated quinolinone
derivatives substituted with N- or C-linked imidazoles
Abstract
This invention concerns compounds of formula 1 the
pharmaceutically acceptable acid addition salts and the
stereochemically isomeric forms thereof, wherein the dotted line
represents an optional bond; X is oxygen or sulfur; -A- is a
bivalent radical of formula; R.sup.1 and R.sup.2 each independently
are hydrogen, hydroxy, halo, cyano, C.sub.1-6alkyl, trihalomethyl,
trihalomethoxy, C.sub.2-6alkenyl, C.sub.1-6alkyloxy,
hydroxyC.sub.1-6alkyloxy, C.sub.1-6alkyloxyC.sub.1-6alkyloxy,
C.sub.1-6alkyloxycarbonyl, aminoC.sub.1-6alkyloxy, mono- or
di(C.sub.1-6alkyl)aminoC.sub.1-6alkyloxy- , Ar, Ar--C.sub.1-6alkyl,
Ar-oxy, Ar--C.sub.1-6alkyloxy; or when on adjacent positions
R.sup.1 and R.sup.2 taken together may form a bivalent radical;
R.sup.3 and R.sup.4 each independently are hydrogen, halo, cyano,
C.sub.1-6alkyl, C.sub.1-6alkyloxy, Ar-oxy, C.sub.1-6alkylthio,
di(C.sub.1-6alkyl)amino, trihalomethyl, trihalomethoxy, or when on
adjacent positions R.sup.3 and R.sup.4 taken together may form a
bivalent radical; R.sup.5 is an imidazolyl substituted with
hydrogen or C.sub.1-6alkyl; R.sup.6 hydrogen, hydroxy, halo, cyano,
optionally substituted C.sub.1-6alkyl, C.sub.1-6alkyloxycarbonyl or
Ar; or a radical of formula --O--R.sup.7, --S--R.sup.8,
--N--R.sup.8R.sup.9; and Ar is optionally substituted phenyl;
having farnesyl transferase inhibiting activity; their preparation,
compositions containing them and their use as a medicine.
Inventors: |
Venet, Marc Gaston; (Le
Mesnil Esnard, FR) ; Angibaud, Patrick Rene;
(Fontaine-Bellenger, FR) ; Ligny, Yannick Aime Eddy;
(Sotteville-les-Rouen, FR) ; Poncelet, Virginie
Sophie; (Le Manoir sur Seine, FR) ; Sanz, Gerard
Charles; (Le Mesnil Esnard, FR) |
Correspondence
Address: |
Philip S. Johnson, Esq.
Johnson & Johnson
One Johnson & Johnson Plaza
New Brunswick
NJ
08933-7003
US
|
Family ID: |
26146222 |
Appl. No.: |
09/725391 |
Filed: |
November 29, 2000 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09725391 |
Nov 29, 2000 |
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09380856 |
Dec 20, 1999 |
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6187786 |
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09380856 |
Dec 20, 1999 |
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PCT/EP98/02357 |
Mar 3, 1998 |
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Current U.S.
Class: |
546/81 ; 544/101;
546/95 |
Current CPC
Class: |
A61P 35/00 20180101;
C07D 471/06 20130101; C07D 455/04 20130101 |
Class at
Publication: |
546/81 ; 546/95;
544/101 |
International
Class: |
C07D 471/00; C07D
491/00; C07D 498/00; C07D 513/00; C07D 515/00 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 10, 1997 |
EP |
97.200.709.0 |
Mar 10, 1997 |
EP |
97.200.708.2 |
Claims
1. A compound of formula (I) 23a pharmaceutically acceptable acid
addition salt or a stereochemically isomeric form thereof, wherein
the dotted line represents an optional bond; X is oxygen or sulfur;
-A- is a bivalent radical of formula
12 --CH.dbd.CH-- (a-1), --CH.sub.2--S-- (a-6),
--CH.sub.2--CH.sub.2-- (a-2), --CH.sub.2--CH.sub.2--S-- (a-7),
--CH.sub.2--CH.sub.2--CH.sub.2-- (a-3), --CH.dbd.N-- (a-8),
--CH.sub.2--O-- (a-4), --N.dbd.N-- (a-9), or
--CH.sub.2--CH.sub.2--O-- (a-5), --CO--NH-- (a-10);
wherein optionally one hydrogen atom may be replaced by
C.sub.1-4alkyl or Ar.sup.1; R.sup.1 and R.sup.2 each independently
are hydrogen, hydroxy, halo, cyano, C.sub.1-6alkyl, trihalomethyl,
trihalomethoxy, C.sub.2-6alkenyl, C.sub.1-6alkyloxy,
hydroxyC.sub.1-6alkyloxy, C.sub.1-6alkyloxyC.sub.1-6alkyloxy,
C.sub.1-6alkyloxycarbonyl, aminoC.sub.1-6alkyloxy, mono- or
di(C.sub.1-6alkyl)aminoC.sub.1-6alkyloxy- , Ar.sup.2,
Ar.sup.2--C.sub.1-6alkyl, Ar.sup.2-oxy,
Ar.sup.2--C.sub.1-6alkyloxy; or when on adjacent positions R.sup.1
and R.sup.2 taken together may form a bivalent radical of
formula
13 --O--CH.sub.2--O-- (b-1), --O--CH.sub.2--CH.sub.2--O-- (b-2),
--O--CH.dbd.CH-- (b-3), --O--CH.sub.2--CH.sub.2-- (b-4),
--O--CH.sub.2--CH.sub.2--CH.su- b.2-- (b-5), or
--CH.dbd.CH--CH.dbd.CH-- (b-6);
R.sup.3 and R.sup.4 each independently are hydrogen, halo, cyano,
C.sub.1-6alkyl, C.sub.1-6alkyloxy, Ar.sup.3-oxy,
C.sub.1-6alkylthio, di(C.sub.1-6alkyl)amino, trihalomethyl,
trihalomethoxy, or when on adjacent positions R.sup.3 and R.sup.4
taken together may form a bivalent radical of formula
14 --O--CH.sub.2--O-- (c-1), --O--CH.sub.2--CH.sub.2--O-- (c-2), or
--CH.dbd.CH--CH.dbd.CH-- (c-3);
R.sup.5 is a radical of formula 24 wherein R.sup.13 is hydrogen,
halo, Ar.sup.4, C.sub.1-6alkyl, hydroxyC.sub.1-6alkyl,
C.sub.1-6alkyloxyC.sub.1- -6alkyl, C.sub.1-6alkyloxy,
C.sub.1-6alkylthio, amino, C.sub.1-6alkyloxycarbonyl,
C.sub.1-6alkylS(O)C.sub.1-6alkyl or
C.sub.1-6alkylS(O).sub.2C.sub.1-6alkyl; R.sup.14 is hydrogen,
C.sub.1-6alkyl or di(C.sub.1 4alkyl)aminosulfonyl; R.sup.6 is
hydrogen, hydroxy, halo, C.sub.1-6alkyl, cyano, haloC.sub.1-6alkyl,
hydroxyC.sub.1-6alkyl, cyanoC.sub.1-6alkyl, aminoC.sub.1-6alkyl,
C.sub.1-6alkyloxyC.sub.1-6alkyl, C.sub.1-6alkylthioC.sub.1-6alkyl,
aminocarbonyl-C.sub.1-6alkyl,
C.sub.1-6alkyloxycarbonylC.sub.1-6alkyl,
C.sub.1-6alkylcarbonylC.sub.1-6alkyl, C.sub.1-6alkyloxycarbonyl,
mono- or di(C.sub.1-6alkyl)aminoC.sub.1-6alkyl, Ar.sup.5,
Ar.sup.5--C.sub.1-6alkyl- oxyC.sub.1-6alkyl; or a radical of
formula
15 --O--R.sup.7 (e-1), --S--R.sup.7 (e-2), or --N--R.sup.8R.sup.9
(e-3),
wherein R.sup.7 is hydrogen, C.sub.1-6alkyl,
C.sub.1-6alkylcarbonyl, Ar.sup.6, Ar.sup.6--C.sub.1-6alkyl,
C.sub.1-6alkyloxycarbonylC.sub.1-6alk- yl, or a radical of formula
-Alk-OR.sup.10 or -Alk-NR.sup.11R.sup.12; R.sup.8 is hydrogen,
C.sub.1-6alkyl, Ar.sup.7 or Ar.sup.7--C.sub.1-6alkyl- ; R.sup.9 is
hydrogen, C.sub.1-6alkyl, C.sub.1-6alkylcarbonyl,
C.sub.1-6alkyloxycarbonyl, C.sub.1-6alkylaminocarbonyl, Ar.sup.8,
Ar.sup.8--C.sub.1-6alkyl, C.sub.1-6alkylcarbonylC.sub.1-6alkyl,
Ar.sup.8-carbonyl, Ar.sup.8--C.sub.1-6alkylcarbonyl,
aminocarbonylcarbonyl, C.sub.1-6alkyloxyC.sub.1-6alkylcarbonyl,
hydroxy, C.sub.1-6alkyloxy, aminocarbonyl,
di(C.sub.1-6alkyl)aminoC.sub.1-6alkylca- rbonyl, amino,
C.sub.1-6alkylamino, C.sub.1-6alkylcarbonylamino, or a radical or
formula -Alk-OR.sup.10 or -Alk-NR.sup.11 R.sup.12; wherein Alk is
C.sub.1-6alkanediyl; R.sup.10 is hydrogen, C.sub.1-6alkyl,
C.sub.1-6alkylcarbonyl, hydroxyC.sub.1-6alkyl, Ar.sup.9 or
Ar.sup.9--C.sub.1-6alkyl; R.sup.11 is hydrogen, C.sub.1-6alkyl,
C.sub.1-6alkylcarbonyl, Ar.sup.10 or Ar.sup.10--C.sub.1-6alkyl;
R.sup.12 is hydrogen, C.sub.1-6alkyl, Ar.sup.11 or
Ar.sup.11--C.sub.1-6alkyl; and Ar.sup.1 to Ar.sup.11 are each
independently selected from phenyl; or phenyl substituted with
halo, C.sub.1-6alkyl, C.sub.1-6alkyloxy or trifluoromethyl.
2. A compound according to claim 1 wherein the dotted line
represents an optional bond; X is O or S; R.sup.1 and R.sup.2 are
each independently selected from hydrogen, halo, C.sub.1-6alkyl,
C.sub.1-6alkyloxy, trihalomethyl or trihalomethoxy; R.sup.3 and
R.sup.4 are each independently selected from hydrogen, halo,
C.sub.1-6alkyl, C.sub.1-6alkyloxy, trihalomethyl or trihalomethoxy;
R.sup.5 a radical of formula (d-1) wherein R.sup.13 is hydrogen or
R.sup.5 is a radical of formula (d-2) wherein R.sup.13 is hydrogen
or C.sub.1-6alkyl and R.sup.14 is hydrogen or C.sub.1-6alkyl;
R.sup.6 is hydrogen, hydroxy, haloC.sub.1-6alkyl,
hydroxyC.sub.1-6alkyl, cyanoC.sub.1-6alkyl,
C.sub.1-6alkyloxycarbonylC.sub.1-6alkyl, or a radical of formula
-NR.sup.8R.sup.9 wherein R.sup.8 is hydrogen or C.sub.1-6alkyl and
R.sup.9 is hydrogen, C.sub.1-6alkyl, C.sub.1-6alkyloxy or
C.sub.1-6alkyloxyC.sub.1-6alkylcarbonyl.
3. A compound according to any of claims 1 to 2 wherein X is
oxygen; the dotted line represents a bond; R.sup.1 is 3-halo;
R.sup.2 is hydrogen; R.sup.3 is 4-halo; R.sup.4 is hydrogen;
R.sup.5 a radical of formula (d-1) wherein R.sup.13 is hydrogen or
R.sup.5 is a radical of formula (d-2) wherein R.sup.13 is hydrogen
and R.sup.14 is C.sub.1-4alkyl; R.sup.6 is hydrogen, halo, hydroxy
or amino; and -A- is (a-1), (a-2) or (a-3).
4. A compound according to claim 1 wherein the compound is
7-(3-chlorophenyl)-9-[(4-chlorophenyl)-1H-imidazol-1-ylmethyl]-2,3-dihydr-
o-1H,5H-benzo[ij]quinolizin-5-one;
7-(3-chlorophenyl)-9-[(4-chlorophenyl)--
1H-imidazol-1-ylmethyl]-1,2-dihydro-4H-pyrrolo[3,2,1-ij]quinoline-4-one;
8-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-6-(3-chlorophe-
nyl)-1,2-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-4-one; or
8-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-6-(3-chlorophe-
nyl)-2,3-dihydro-1H,5H-benzo[ij]quinolizin-5-one; a stereoisomeric
form or a pharmaceutically acceptable acid addition salt
thereof.
5. A pharmaceutical composition comprising a pharmaceutically
acceptable carrier, and as active ingredient a therapeutically
effective amount of a compound as described in any one of claims 1
to 4.
6. A process for preparing a pharmaceutical composition as claimed
in claim 5 wherein a therapeutically effective amount of a compound
as claimed in any one of claims 1 to 4 is intimately mixed with a
pharmaceutically acceptable carrier.
7. A compound of formula (VI) 25an acid addition salt or a
stereochemically isomeric form thereof, wherein X, R.sup.1, R.sup.2
and -A- are as defined in claim 1.
8. A compound of formula (II) 26an acid addition salt or a
stereochemically isomeric form thereof, wherein the dotted line
represents an optional bond; X, R.sup.1, R.sup.2, R.sup.3, R.sup.4
and -A- are as defined in claim 1.
9. A compound according to any one of claims 1 to 4 for use as a
medicine.
10. A process for preparing a compound as claimed in claim 1,
wherein a) an intermediate ketone of formula (II) is reacted with
an intermediate of formula (III-1) or (III-2) in the presence of a
suitable strong base and in the presence an appropriate
silanederivative, optionally followed by removal of a protective
group PG; 27b) compounds of formula (I-a), defined as compounds of
formula (I) wherein R.sup.6 is hydroxy, are converted to compounds
of formula (I-c) wherein R.sup.6 is halo, optionally followed by
treatment with an intermediate of formula H--NR.sup.8R.sup.9
yielding compounds of formula (I-d); 28c) an intermediate of
formula (XVIII) is N-alkylated with an intermediate of formula
(XVII) in a reaction-inert solvent and, optionally in the presence
of a suitable base; 29d) an intermediate of formula (XIX) is
N-alkylated with a compound of formula (XVI); 30 wherein in the
above reaction schemes the dotted line and the radicals X, R.sup.1,
R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.8, R.sup.9 and
R.sup.13 and -A- are as defined in claim 1, W is an appropriate
leaving group and Y is carbon or sulfur; e) or, compounds of
formula (I) are converted into each other following art-known
transformation reactions; or if desired; a compound of formula (I)
is converted into a pharmaceutically acceptable acid addition salt,
or conversely, an acid addition salt of a compound of formula (I)
is converted into a free base form with alkali; and, if desired,
preparing stereochemically isomeric forms thereof.
11. A process for preparing a compound of formula (VI) as claimed
in claim 7 wherein an intermediate of formula (IV) is cyclized in
the presence of polyphosphoric acid (PPA), 31wherein in the above
reaction schemes the radicals X, R.sup.1, R.sup.2 and -A- are as
defined in claim 1; or, compounds of formula (VI) are converted
into each other following art-known transformation reactions; or if
desired; a compound of formula (VI) is converted into a
pharmaceutically acceptable acid addition salt, or conversely, an
acid addition salt of a compound of formula (VI) is converted into
a free base form with alkali; and, if desired, preparing
stereochemically isomeric forms thereof.
12. A process for preparing an intermediate compound of formula
(II) as claimed in claim 8 wherein a) an intermediate of formula
(VI) is treated with an intermediate of formula (V) in the presence
of polyphosphoric acid (PPA); 32b) or, an intermediate of formula
(IV) with an intermediate of formula (V) in the presence of
polyphosphoric acid (PPA); 33c) intermediates of formula (II-a),
defined as intermediates of formula (II) wherein the dotted line
does not represent a bond, can be converted into intermediates of
formula (II-b), defined as an intermediate of formula (II) wherein
the dotted line represents a bond, by oxidation 34 wherein in the
above reaction schemes the radicals X, R.sup.1, R.sup.2, R.sup.3,
R.sup.4 and -A- are as defined in claim 1; d) or, compounds of
formula (II-a) are converted into each other following art-known
transformation reactions; or if desired; a compound of formula
(II-a) is converted into a pharmaceutically acceptable acid
addition salt, or conversely, an acid addition salt of a compound
of formula (II-a) is converted into a free base form with alkali;
and, if desired, preparing stereochemically isomeric forms thereof.
Description
[0001] The present invention is concerned with novel 1,8-annelated
2-quinolinone derivatives, the preparation thereof, pharmaceutical
compositions comprising said novel compounds and the use of these
compounds as a medicine as well as methods of treatment by
administering said compounds.
[0002] Oncogenes frequently encode protein components of signal
transduction pathways which lead to stimulation of cell growth and
mitogenesis. Oncogene expression in cultured cells leads to
cellular transformation, characterized by the ability of cells to
grow in soft agar and the growth of cells as dense foci lacking the
contact inhibition exhibited by non-transformed cells. Mutation
and/or overexpression of certain oncogenes is frequently associated
with human cancer. A particular group of oncogenes is known as ras
which have been identified in mammals, birds, insects, mollusks,
plants, fungi and yeasts. The family of mammalian ras oncogenes
consists of three major members ("isoforms"): H-ras, K-ras and
N-ras oncogenes. These ras oncogenes code for highly related
proteins generically known as p21.sup.ras. Once attached to plasma
membranes, the mutant or oncogenic forms of p21.sup.ras will
provide a signal for the transformation and uncontrolled growth of
malignant tumor cells. To acquire this transforming potential, the
precursor of the p21.sup.ras oncoprotein must undergo an
enzymatically catalyzed farnesylation of the cysteine residue
located in a carboxyl-terminal tetrapeptide. Therefore, inhibitors
of the enzyme that catalyzes this modification, farnesyl protein
transferase, will prevent the membrane attachment of p21.sup.ras
and block the aberrant growth of ras-transformed tumors. Hence, it
is generally accepted in the art that farnesyl transferase
inhibitors can be very useful as anticancer agents for tumors in
which ras contributes to transformation.
[0003] Since mutated oncogenic forms of ras are frequently found in
many human cancers, most notably in more than 50% of colon and
pancreatic carcinomas (Kohl et al., Science, vol 260, 1834-1837,
1993), it has been suggested that farnesyl tranferase inhibitors
can be very useful against these types of cancer.
[0004] In EP-0,371,564 there are described (1H-azol-1-ylmethyl)
substituted quinoline and quinolinone derivatives which suppress
the plasma elimination of retinoic acids. Some of these compounds
also have the ability to inhibit the formation of androgens from
progestines and/or inhibit the action of the aromatase enzyme
complex.
[0005] Unexpectedly, it has been found that the present novel
compounds, all having a phenyl substituent on the 4-position of the
1,8-annelated 2-quinolinone-moiety bearing a nitrogen- or
carbon-linked imidazole, show farnesyl protein transferase
inhibiting activity.
[0006] The present invention concerns compounds of formula 2
[0007] the pharmaceutically acceptable acid addition salts and the
stereochemically isomeric forms thereof, wherein
[0008] the dotted line represents an optional bond;
[0009] X is oxygen or sulfur;
[0010] -A- is a bivalent radical of formula
1 --CH.dbd.CH-- (a-1), --CH.sub.2--S-- (a-6),
--CH.sub.2--CH.sub.2-- (a-2), --CH.sub.2--CH.sub.2--S-- (a-7),
--CH.sub.2--CH.sub.2--CH.sub.2-- (a-3), --CH.dbd.N-- (a-8),
--CH.sub.2--O-- (a-4), --N.dbd.N-- (a-9), or
--CH.sub.2--CH.sub.2--O-- (a-5), --CO--NH-- (a-10);
[0011] wherein optionally one hydrogen atom may be replaced by
C.sub.1-4alkyl or Ar.sup.1;
[0012] R.sup.1 and R.sup.2 each independently are hydrogen,
hydroxy, halo, cyano, C.sub.1-6alkyl, trihalomethyl,
trihalomethoxy, C.sub.2-6alkenyl, C.sub.1-6alkyloxy,
hydroxyC.sub.1-6alkyloxy, C.sub.1-6alkyloxyC.sub.1-6al- kyloxy,
C.sub.1-6alkyloxycarbonyl, aminoC.sub.1-6alkyloxy, mono- or
di(C.sub.1-6alkyl)aminoC.sub.1-6alkyloxy, Ar.sup.2,
Ar.sup.2--C.sub.1-6alkyl, Ar.sup.2-oxy,
Ar.sup.2--C.sub.1-6alkyloxy; or when on adjacent positions R.sup.1
and R.sup.2 taken together may form a bivalent radical of
formula
2 --O--CH.sub.2--O-- (b-1), --O--CH.sub.2--CH.sub.2--O-- (b-2),
--O--CH.dbd.CH-- (b-3), --O--CH.sub.2--CH.sub.2-- (b-4),
--O--CH.sub.2--CH.sub.2--CH.su- b.2-- (b-5), or
--CH.dbd.CH--CH.dbd.CH-- (b-6);
[0013] R.sup.3 and R.sup.4 each independently are hydrogen, halo,
cyano, C.sub.1-6alkyl, C.sub.1-6alkyloxy, Ar.sup.3-oxy,
C.sub.1-6alkylthio, di(C.sub.1-6alkyl)amino, trihalomethyl,
trihalomethoxy, or when on adjacent positions R.sup.3 and R.sup.4
taken together may form a bivalent radical of formula
3 --O--CH.sub.2--O-- (c-1), --O--CH.sub.2--CH.sub.2--O-- (c-2), or
--CH.dbd.CH--CH.dbd.CH-- (c-3);
[0014] R.sup.5 is a radical of formula 3
[0015] wherein
[0016] R.sup.13 is hydrogen, halo, Ar.sup.4, C.sub.1-6alkyl,
hydroxyC.sub.1-6alkyl, C.sub.1-6alkyloxyC.sub.1-6alkyl,
C.sub.1-6alkyloxy, C.sub.1-6alkylthio, amino,
C.sub.1-6alkyloxycarbonyl, C.sub.1-6alkylS(O)C.sub.1-6alkyl or
C.sub.1-6alkylS(O).sub.2C.sub.1-6alky- l;
[0017] R.sup.14 is hydrogen, C.sub.1-6alkyl or
di(C.sub.1-4alkyl)aminosulf- onyl;
[0018] R.sup.6 is hydrogen, hydroxy, halo, C.sub.1-6alkyl, cyano,
haloC.sub.1-6alkyl, hydroxyC.sub.1-6alkyl, cyanoC.sub.1-6alkyl,
aminoC.sub.1-6alkyl, C.sub.1-6alkyloxyC.sub.1-6alkyl,
C.sub.1-6alkylthioC.sub.1-6alkyl, aminocarbonylC.sub.1-6alkyl,
C.sub.1-6alkyloxycarbonylC.sub.1-6alkyl,
C.sub.1-6alkylcarbonyl-C.sub.1-6- alkyl, C.sub.1-6alkyloxycarbonyl,
mono- or di(C.sub.1-6alkyl)aminoC.sub.1-- 6alkyl, Ar.sup.5,
Ar.sup.5--C.sub.1-6alkyloxyC.sub.1-6alkyl; or a radical of
formula
4 --O--R.sup.7 (e-1), --S--R.sup.7 (e-2), --N--R.sup.8R.sup.9
(e-3),
[0019] wherein
[0020] R.sup.7 is hydrogen, C.sub.1-6alkyl, C.sub.1-6alkylcarbonyl,
Ar.sup.6, Ar.sup.6--C.sub.1-6alkyl,
C.sub.1-6alkyloxycarbonylC.sub.1-6alk- yl, or a radical of formula
-Alk-OR.sup.10 or -Alk-NR.sup.11R.sup.12;
[0021] R.sup.8 is hydrogen, C.sub.1-6alkyl, Ar.sup.7 or
Ar.sup.7--C.sub.1-6alkyl;
[0022] R.sup.9 is hydrogen, C.sub.1-6alkyl, C.sub.1-6alkylcarbonyl,
C.sub.1-6alkyloxycarbonyl, C.sub.1-6alkylaminocarbonyl, Ar.sup.8,
Ar.sup.8--C.sub.1-6alkyl, C.sub.1-6alkylcarbonyl-C.sub.1-6alkyl,
Ar.sup.8-carbonyl, Ar.sup.8--C.sub.1-6alkylcarbonyl,
aminocarbonylcarbonyl, C.sub.1-6alkyloxyC.sub.1-6alkylcarbonyl,
hydroxy, C.sub.1-6alkyloxy, aminocarbonyl,
di(C.sub.1-6alkyl)aminoC.sub.1-6alkylca- rbonyl, amino,
C.sub.1-6alkylamino, C.sub.1-6alkylcarbonylamino, or a radical or
formula -Alk-OR.sup.10 or -Alk-NR.sup.11R.sup.12;
[0023] wherein Alk is C.sub.1-6alkanediyl;
[0024] R.sup.10 is hydrogen, C.sub.1-6alkyl,
C.sub.1-6alkylcarbonyl, hydroxyC.sub.1-6alkyl, Ar.sup.9 or
Ar.sup.9--C.sub.1-6alkyl;
[0025] R.sup.11 is hydrogen, C.sub.1-6alkyl,
C.sub.1-6alkylcarbonyl, Ar.sup.10 or Ar.sup.10--C.sub.1-6alkyl;
[0026] R.sup.12 is hydrogen, C.sub.1-6alkyl, Ar.sup.11 or
Ar.sup.11--C.sub.1-6alkyl; and
[0027] Ar.sup.1 to Ar.sup.11 are each independently selected from
phenyl; or phenyl substituted with halo, C.sub.1-6alkyl,
C.sub.1-6alkyloxy or trifluoromethyl.
[0028] R.sup.13 may also be bound to one of the nitrogen atoms in
the imidazole ring of formula (d-1). In that case the meaning of
R.sup.13 when bound to the nitrogen is limited to hydrogen,
Ar.sup.4, C.sub.1-6alkyl, hydroxyC.sub.1-6alkyl,
C.sub.1-6alkyloxyC.sub.1-6alkyl, C.sub.1-6alkyloxycarbonyl,
C.sub.1-6alkylS(O)--C.sub.1-6alkyl or
C.sub.1-6alkylS(O).sub.2C.sub.1-6alkyl.
[0029] As used in the foregoing definitions and hereinafter, halo
is generic to fluoro, chloro, bromo and iodo; C.sub.1-4alkyl
defines straight and branched chain saturated hydrocarbon radicals
having from 1 to 4 carbon atoms such as, e.g. methyl, ethyl,
propyl, butyl, 1-methylethyl, 2-methylpropyl and the like;
C.sub.1-6alkyl includes C.sub.1-4alkyl and the higher homologues
thereof having 5 to 6 carbon atoms such as, for example, pentyl,
2-methylbutyl, hexyl, 2-methylpentyl and the like;
C.sub.1-6alkanediyl defines bivalent straight and branched chained
saturated hydrocarbon radicals having from 1 to 6 carbon atoms,
such as, for example, methylene, 1,2-ethanediyl, 1,3-propanediyl,
1,4-butanediyl, 1,5-pentanediyl, 1,6-hexanediyl and the branched
isomers thereof; C.sub.2-6alkenyl defines straight and branched
chain hydrocarbon radicals containing one double bond and having
from 2 to 6 carbon atoms such as, for example, ethenyl, 2-propenyl,
3-butenyl, 2-pentenyl, 3-pentenyl, 3-methyl-2-butenyl, and the
like. The term "S(O)" refers to a sulfoxide and "S(O).sub.2" to a
sulfon.
[0030] The pharmaceutically acceptable acid addition salts as
mentioned hereinabove are meant to comprise the therapeutically
active non-toxic acid addition salt forms which the compounds of
formula (I) are able to form. The compounds of formula (I) which
have basic properties can be converted in their pharmaceutically
acceptable acid addition salts by treating said base form with an
appropriate acid. Appropriate acids comprise, for example,
inorganic acids such as hydrohalic acids, e.g. hydrochloric or
hydrobromic acid; sulfuric; nitric; phosphoric and the like acids;
or organic acids such as, for example, acetic, propanoic,
hydroxyacetic, lactic, pyruvic, oxalic, malonic, succinic (i.e.
butanedioic acid), maleic, fumaric, malic, tartaric, citric,
methanesulfonic, ethanesulfonic, benzenesulfonic,
p-toluenesulfonic, cyclamic, salicylic, p-aminosalicylic, pamoic
and the like acids.
[0031] The term acid addition salts also comprises the hydrates and
the solvent addition forms which the compounds of formula (I) are
able to form. Examples of such forms are e.g. hydrates, alcoholates
and the like.
[0032] The term stereochemically isomeric forms of compounds of
formula (I), as used hereinbefore, defines all possible compounds
made up of the same atoms bonded by the same sequence of bonds but
having different three-dimensional structures which are not
interchangeable, which the compounds of formula (I) may possess.
Unless otherwise mentioned or indicated, the chemical designation
of a compound encompasses the mixture of all possible
stereochemically isomeric forms which said compound may possess.
Said mixture may contain all diastereomers and/or enantiomers of
the basic molecular structure of said compound. All
stereochemically isomeric forms of the compounds of formula (I)
both in pure form or in admixture with each other are intended to
be embraced within the scope of the present invention.
[0033] Some of the compounds of formula (I) may also exist in their
tautomeric forms. Such forms although not explicitly indicated in
the above formula are intended to be included within the scope of
the present invention.
[0034] Wherever -A- is a bivalent radical of formula (a-4), (a-5),
(a-6), (a-7) or (a-8) the CH.sub.2 or CH moiety in said bivalent
radical is preferably connected to the nitrogen atom of the
2-quinolinone-moiety of the compounds of formula (I) or the
intermediates of formula (II), (IV), (VI) and (VII).
[0035] Whenever used hereinafter, the term "compounds of formula
(I)" is meant to include also the pharmaceutically acceptable acid
addition salts and all stereoisomeric forms.
[0036] A group of interesting compounds consists of those compounds
of formula (I) wherein one or more of the following restrictions
apply:
[0037] a) the dotted line represents an optional bond;
[0038] b) X is O or S;
[0039] c) R.sup.1 and R.sup.2 are each independently selected from
hydrogen, halo, C.sub.1-6alkyl, C.sub.1-6alkyloxy, trihalomethyl or
trihalomethoxy; in particular hydrogen, halo or C.sub.1-4alkyl;
[0040] d) R.sup.3 and R.sup.4 are each independently selected from
hydrogen, halo, C.sub.1-6alkyl, C.sub.1-6alkyloxy, trihalomethyl or
trihalomethoxy; in particular hydrogen, halo, C.sub.1-4alkyl or
C.sub.1-4alkyloxy;
[0041] e) R.sup.5 is a radical of formula (d-1) wherein R.sup.13 is
hydrogen or C.sub.1-6alkyl; or R.sup.5 is a radical of formula
(d-2) wherein R.sup.13 is hydrogen or C.sub.1-6alkyl and R.sup.14
is hydrogen or C.sub.1-6alkyl;
[0042] f) R.sup.6 is hydrogen, hydroxy, haloC.sub.1-6alkyl,
hydroxyC.sub.1-6alkyl, cyanoC.sub.1-6alkyl,
C.sub.1-6alkyloxycarbonylC.su- b.1-6alkyl, or a radical of formula
--NR.sup.8R.sup.9 wherein R.sup.8 is hydrogen or C.sub.1-6alkyl and
R.sup.9 is hydrogen, C.sub.1-6alkyl, C.sub.1-6alkyloxy,
C.sub.1-6alkyloxyC.sub.1-6alkylcarbonyl; in particular R.sup.6 is
hydrogen, hydroxy, halo or a amino;
[0043] g) -A- is (a-1), (a-2), (a-3), (a-4), (a-5), (a-8), (a-9) or
(a-10).
[0044] A particular group of compounds consists of those compounds
of formula (I) wherein the dotted line represents a bond; X is O or
S; R.sup.2 is hydrogen and R.sup.1 is halo, preferably chloro,
especially 3-chloro; R.sup.4 is hydrogen and R.sup.3 is halo,
preferably chloro, especially 4-chloro; R.sup.5 is a radical of
formula (d-1) wherein R.sup.13 is hydrogen or C.sub.1-4alkyl; and
R.sup.6 is hydrogen.
[0045] Another particular group of compounds consists of those
compounds of formula (I) wherein the dotted line represent a bond;
X is O or S; R.sup.2 is hydrogen and R.sup.1 is halo, preferably
chloro, especially 3-chloro; and R.sup.4 is hydrogen and R.sup.3 is
halo, preferably chloro, especially 4-chloro; R.sup.5 is a radical
of formula (d-2) wherein R.sup.13 is hydrogen or C.sub.1-4alkyl and
R.sup.14 is hydrogen or C.sub.1-4alkyl; R.sup.6 is hydrogen,
hydroxy, halo or amino.
[0046] Preferred compounds are those compounds of formula (I)
wherein the dotted line represents a bond; X is oxygen; R.sup.1 is
3-chloro; R.sup.2 is hydrogen; R.sup.3 is 4-chloro; R.sup.4 is
hydrogen; R.sup.5 is a radical of formula (d-1) wherein R.sup.13 is
hydrogen or C.sub.1-4alkyl; R.sup.6 is hydrogen, and -A- is (a-1),
(a-2) or (a-3).
[0047] Other preferred compounds are those compounds of formula (I)
wherein the dotted line represent a bond; X is oxygen; R.sup.1 is
3-chloro; R.sup.2 is hydrogen; R.sup.3 is 4-chloro; R.sup.4 is
hydrogen; R.sup.5 is a radical of formula (d-2) wherein R.sup.13 is
hydrogen and R.sup.14 is C.sub.1-4alkyl; R.sup.6 is amino; and -A-
is (a-1), (a-2) or (a-3).
[0048] The most preferred compounds of formula (I) are
[0049]
7-(3-chlorophenyl)-9-[(4-chlorophenyl)-1H-imidazol-1-ylmethyl]-2,3--
dihydro-1H,5H-benzo[ij]quinolizin-5-one,
[0050]
7-(3-chlorophenyl)-9-[(4-chlorophenyl)-1H-imidazol-1-ylmethyl]-1,2--
dihydro-4H-pyrrolo[3,2,1-ij]quinoline-4-one,
[0051]
8-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-6-(3-chl-
orophenyl)-1,2-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-4-one, and
[0052]
8-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-6-(3-chl-
orophenyl)-2,3-dihydro-1H,5H-benzo[ij]quinolizin-5-one; the
stereoisomeric forms and the pharmaceutically acceptable acid
addition salts thereof.
[0053] The compounds of formula (I), wherein R.sup.6 is hydroxy and
R.sup.5 is a radical of formula (d-2) wherein R.sup.14 is
C.sub.1-6alkyl, said compounds being referred to as compounds of
formula (I-a-1) may be prepared by reacting an intermediate ketone
of formula (II) with a intermediate of formula (III-1). Said
reaction requires the presence of a suitable strong base, such as,
for example, butyl lithium in an appropriate solvent, such as, for
example, tetrahydrofuran, and the presence of an appropriate
silanederivative, such as, for example, triethylchlorosilane.
During the work-up procedure an intermediate silane derivative is
hydrolyzed. Other procedures with protective groups analogous to
silanederivatives can also be applied. 4
[0054] Also, the compounds of formula (I), wherein R.sup.6 is
hydroxy and R.sup.5 is a radical of formula (d-2) wherein R.sup.14
is hydrogen, said compounds being referred to as compounds of
formula (I-a-2) may be prepared by reacting an intermediate ketone
of formula (II) with a intermediate of formula (III-2), wherein PG
is a protective group such as, for example, a sulfonyl group, e.g.
a dimethylamino sulfonyl group, which can be removed after the
addition reaction. Said reaction is conducted analogously as for
the preparation of compounds of formula (I-a-1), followed by
removal of the protecting group PG, yielding compounds of formula
(I-a-2).
[0055] Compounds of formula (I-g), defined as compounds of formula
(I) wherein R.sup.5 represents a radical of formula (d-1), can be
prepared by N-alkylating an intermediate of formula (XVIII) with an
intermediate of formula (XVII), wherein W is an appropriate leaving
group such as, for example, chloro, bromo, methanesulfonyloxy or
benzenesulfonyloxy. The reaction can be performed in a
reaction-inert solvent such as, for example, acetonitrile, and
optionally in the presence of a suitable base such as, for example,
sodium carbonate, potassium carbonate or triethylamine. Stirring
may enhance the rate of the reaction. The reaction may conveniently
be carried out at a temperature ranging between room temperature
and the reflux temperature of the reaction mixture. 5
[0056] Also, compounds of formula (I-g) can be prepared by
N-alkylating an intermediate of formula (XIX), wherein Y is carbon
or sulfur, such as, for example, a 1,1'-carbonyl-diimidazole, with
an intermediate of formula (XVI). 6
[0057] Said reaction may conveniently be conducted in a
reaction-inert solvent, such as, e.g. tetrahydrofuran, optionally
in the presence of a base, such as sodium hydride, and at a
temperature ranging between room temperature and the reflux
temperature of the reaction mixture.
[0058] Compounds of formula (I-g) may also be prepared by reacting
an intermediate of formula (XVII) with ammonia and subsequent
treatment with isothiocyanate as described in EP-0,293,978 page 12,
line 33 to page 13, line 20.
[0059] The compounds of formula (I-a) can be converted to compounds
of formula (I-b), defined as a compound of formula (I) wherein
R.sup.6 is hydrogen, by submitting the compounds of formula (I-a)
to appropriate reducing conditions, such as, e.g. stirring in
acetic acid in the presence of formamide. 7
[0060] Further, compounds of formula (I-a) can be converted to
compounds of formula (I-c) wherein R.sup.6 is halo, by reacting the
compounds of formula (I-a) with a suitable halogenating agent, such
as, e.g. thionyl chloride or phosphorus tribromide. Successively,
the compounds of formula (I-c) can be treated with a reagent of
formula H--NR.sup.8R.sup.9 in a reaction-inert solvent, thereby
yielding compounds of formula (I-d). 8
[0061] A compound of formula (I-f), defined as a compound of
formula (I) wherein X is sulfur, may be prepared by reacting the
corresponding compound of formula (I-e), defined as a compound of
formula (I) wherein X is oxygen, with a reagent like phosphorus
pentasulfide or Lawesson's reagent in a suitable solvent such as,
for example, pyridine. 9
[0062] An intermediate of formula (II-b), defined as an
intermediate of formula (II) wherein the dotted line represents a
bond, can be prepared by oxidizing an intermediate of formula
(II-a), defined as intermediates of formula (II) wherein the dotted
line does not represent a bond, following art-known oxidation
methods such as, for example, treatment with bromine in an
appropriate solvent such as, e.g. bromobenzene, or treatment with
iodine in the presence of acetic acid and potassium acetate. 10
[0063] Said oxidation reaction can give rise to side-products
wherein the bivalent radical -A- is oxidized. For instance,
oxidation of intermediates of formula (II-a) wherein -A- is (a-2)
may give intermediates of formula (II-b) wherein -A- is (a-1).
[0064] Intermediates of formula (XVI) wherein R.sup.6 is hydrogen,
said compounds being represented by formula (XVI-a), can be
prepared by reacting intermediates of formula (II) with an
appropriate reducing agent such as, e.g. sodium borohydride, in a
suitable solvent such as, e.g. methanol. Optionally, intermediates
of formula (XVI) may be converted to intermediates of formula
(XVII) wherein R.sup.6 is hydrogen, said compounds being
represented by formula (XVII-a), by treating (XVI-a) with a
suitable reagent such as, e.g. methanesulfonyloxy chloride, or a
halogenating reagent such as, e.g. POCl.sub.3 or SOCl.sub.2. 11
[0065] Intermediates of formula (II-a) can be prepared by reacting
intermediates of formula (IV) with intermediates of formula (V) in
the presence of polyphosphoric acid (PPA), at a temperature ranging
between room temperature and the reflux temperature of the reaction
mixture. Optionally said reaction may be performed in a
reaction-inert solvent. 12
[0066] Alternatively, an intermediate of formula (II-a) can be made
in a two-step synthesis by cyclizing an intermediate of formula
(IV) in the presence of polyphosphoric acid (PPA) and subsequent
treating the thus obtained intermediate (VI) with an intermediate
of formula (VII) in the presence of PPA. Said two-step synthesis
may be conducted in a "one-pot" synthesis or, if desired,
intermediates of formula (VI) may be isolated and purified before
reaction with intermediates of formula (V). 13
[0067] Intermediates of formula (IV) can be prepared by treating
intermediates of formula (VIII), wherein X is oxygen or sulfur and
Z is hydroxy or halo, with an intermediate of formula (VII) in a
reaction-inert solvent such as, e.g. dichloromethane, and in the
presence of a base such as, e.g. triethylamine, to pick up the acid
liberated during the reaction. 14
[0068] Intermediates of formula (II-b-1), being intermediates of
formula (II-b) wherein X is oxygen and -A'- is a bivalent radical
of formula (a-4) or (a-5), can be prepared starting from an
intermediate of formula (IX). Said intermediates (IX) are
conveniently prepared by protecting the corresponding art-known
ketones. Intermediates of formula (IX) are stirred with
intermediates of formula (X) in the presence of a base such as
sodium hydroxide, in an appropriate solvent, such as an alcohol,
e.g. methanol. The thus obtained intermediates of formula (XI) are
converted to intermediates of formula (XII) in the presence of a
suitable reagent such as, an acid, e.g. TiCl.sub.3, in the presence
of water; or by hydrogenation under acidic conditions in the
presence of a suitable catalyst e.g. platinum on carbon; and by
subsequent treatment with acetic anhydride. Intermediates of
formula (XII) undergo ring closure in the presence of a base such
as, for example, potassium tert-butoxide, and subsequently
hydrolysis, yielding intermediates of formula (XIII). After
conversion of the methoxy group of intermediates of formula (XVIII)
into hydroxy, by treatment with a suitable agent such as, e.g.
borontribromide, the intermediates of formula (XIV) are treated
with an intermediate of formula (XV), wherein A' is a bivalent
radical of formula (a-4) or (a-5), thereby yielding intermediates
of formula (II-b-1). 15
[0069] The compounds of formula (I) and some of the intermediates
have at least one stereogenic center in their structure. This
stereogenic center may be present in a R or a S configuration.
[0070] The compounds of formula (I) as prepared in the hereinabove
described processes are generally racemic mixtures of enantiomers
which can be separated from one another following art-known
resolution procedures. The racemic compounds of formula (I) may be
converted into the corresponding diastereomeric salt forms by
reaction with a suitable chiral acid. Said diastereomeric salt
forms are subsequently separated, for example, by selective or
fractional crystallization and the enantiomers are liberated
therefrom by alkali. An alternative manner of separating the
enantiomeric forms of the compounds of formula (I) involves liquid
chromatography using a chiral stationary phase. Said pure
stereochemically isomeric forms may also be derived from the
corresponding pure stereochemically isomeric forms of the
appropriate starting materials, provided that the reaction occurs
stereospecifically. Preferably if a specific stereoisomer is
desired, said compound will be synthesized by stereospecific
methods of preparation. These methods will advantageously employ
enantiomerically pure starting materials.
[0071] The compounds of formula (I), the pharmaceutically
acceptable acid addition salts and stereoisomeric forms thereof
have valuable pharmacological properties in that they inhibit
farnesyl protein transferase (FPTase), as can be evidenced by the
results obtained in the pharmacological examples C-1 and C-2.
[0072] Furthermore, it is believed that the compounds of formula
(I) wherein R.sup.5 is a radical of formula (d-2) can also inhibit
geranylgeranyltransferase (GGTase).
[0073] This invention provides a method for inhibiting the abnormal
growth of cells, including transformed cells, by administering an
effective amount of a compound of the invention. Abnormal growth of
cells refers to cell growth independent of normal regulatory
mechanisms (e.g. loss of contact inhibition). This includes the
abnormal growth of: (1) tumor cells (tumors) expressing an
activated ras oncogene; (2) tumor cells in which the ras protein is
activated as a result of oncogenic mutation of another gene; (3)
benign and malignant cells of other proliferative diseases in which
aberrant ras activation occurs. Furthermore, it has been suggested
in literature that ras oncogenes not only contribute to the growth
of of tumors in vivo by a direct effect on tumor cell growth but
also indirectly, i.e. by facilitating tumor-induced angiogenesis
(Rak. J. et al, Cancer Research, 55, 4575-4580, 1995). Hence,
pharmacologically targetting mutant ras oncogenes could conceivably
suppress solid tumor growth in vivo, in part, by inhibiting
tumor-induced angiogenesis.
[0074] This invention also provides a method for inhibiting tumor
growth by administering an effective amount of a compound of the
present invention, to a subject, e.g. a mammal (and more
particularly a human) in need of such treatment. In particular,
this invention provides a method for inhibiting the growth of
tumors expressing an activated ras oncogene by the administration
of an effective amount of the compounds of the present invention.
Examples of tumors which may be inhibited, but are not limited to,
lung cancer (e.g. adenocarcinoma), pancreatic cancers (e.g.
pancreatic carcinoma such as, for example exocrine pancreatic
carcinoma), colon cancers (e.g. colorectal carcinomas, such as, for
example, colon adenocarcinoma and colon adenoma), hematopoietic
tumors of lymphoid lineage (e.g. acute lymphocytic leukemia, B-cell
lymphoma, Burkitt's lymphoma), myeloid leukemias (for example,
acute myelogenous leukemia (AML)), thyroid follicular cancer,
myelodysplastic syndrome (MDS), tumors of mesenchymal origin (e.g.
fibrosarcomas and rhabdomyosarcomas), melanomas, teratocarcinomas,
neuroblastomas, gliomas, benign tumor of the skin (e.g.
keratoacanthomas), breast carcinoma, kidney carninoma, ovary
carcinoma, bladder carcinoma and epidermal carcinoma.
[0075] This invention may also provide a method for inhibiting
proliferative diseases, both benign and malignant, wherein ras
proteins are aberrantly activated as a result of oncogenic mutation
in genes, i.e. the ras gene itself is not activated by mutation to
an oncogenic form, with said inhibition being accomplished by the
administration of an effective amount of the compounds described
herein, to a subject in need of such a treatment. For example, the
benign proliferative disorder neurofibromatosis, or tumors in which
ras is activated due to mutation or overexpression of tyrosine
kinase oncogenes may be inhibited by the compounds of this
invention.
[0076] Hence, the present invention discloses the compounds of
formula (I) for use as a medicine as well as the use of these
compounds of formula (I) for the manufacture of a medicament for
treating one or more of the above mentioned conditions.
[0077] In view of their useful pharmacological properties, the
subject compounds may be formulated into various pharmaceutical
forms for administration purposes.
[0078] To prepare the pharmaceutical compositions of this
invention, an effective amount of a particular compound, in base or
acid addition salt form, as the active ingredient is combined in
intimate admixture with a pharmaceutically acceptable carrier,
which carrier may take a wide variety of forms depending on the
form of preparation desired for administration. These
pharmaceutical compositions are desirably in unitary dosage form
suitable, preferably, for administration orally, rectally,
percutaneously, or by parenteral injection. For example, in
preparing the compositions in oral dosage form, any of the usual
pharmaceutical media may be employed, such as, for example, water,
glycols, oils, alcohols and the like in the case of oral liquid
preparations such as suspensions, syrups, elixirs and solutions; or
solid carriers such as starches, sugars, kaolin, lubricants,
binders, disintegrating agents and the like in the case of powders,
pills, capsules and tablets. Because of their ease in
administration, tablets and capsules represent the most
advantageous oral dosage unit form, in which case solid
pharmaceutical carriers are obviously employed. For parenteral
compositions, the carrier will usually comprise sterile water, at
least in large part, though other ingredients, to aid solubility
for example, may be included. Injectable solutions, for example,
may be prepared in which the carrier comprises saline solution,
glucose solution or a mixture of saline and glucose solution.
Injectable suspensions may also be prepared in which case
appropriate liquid carriers, suspending agents and the like may be
employed. In the compositions suitable for percutaneous
administration, the carrier optionally comprises a penetration
enhancing agent and/or a suitable wetting agent, optionally
combined with suitable additives of any nature in minor
proportions, which additives do not cause a significant deleterious
effect to the skin. Said additives may facilitate the
administration to the skin and/or may be helpful for preparing the
desired compositions. These compositions may be administered in
various ways, e.g., as a transdermal patch, as a spot-on, as an
ointment. It is especially advantageous to formulate the
aforementioned pharmaceutical compositions in dosage unit form for
ease of administration and uniformity of dosage. Dosage unit form
as used in the specification and claims herein refers to physically
discrete units suitable as unitary dosages, each unit containing a
predetermined quantity of active ingredient calculated to produce
the desired therapeutic effect in association with the required
pharmaceutical carrier. Examples of such dosage unit forms are
tablets (including scored or coated tablets), capsules, pills,
powder packets, wafers, injectable solutions or suspensions,
teaspoonfuls, tablespoonfuls and the like, and segregated multiples
thereof.
[0079] Those skilled in the art could easily determine the
effective amount from the test results presented hereinafter. In
general it is contemplated that an effective amount would be from
0.01 mg/kg to 100 mg/kg body weight, and in particular from 0.05
mg/kg to 10 mg/kg body weight. It may be appropriate to administer
the required dose as two, three, four or more sub-doses at
appropriate intervals throughout the day. Said sub-doses may be
formulated as unit dosage forms, for example, containing 0.5 to 500
mg, and in particular 1 mg to 200 mg of active ingredient per unit
dosage form.
[0080] The following examples are provided for purposes of
illustration.
[0081] Experimental Part
[0082] Hereinafter "ACN" means acetonitrile, "THF" means
tetrahydrofuran, "DIPE" means diisopropylether, "DCM" means
dichloromethane and "DMF" means N,N-dimethylformamide.
[0083] Of some compounds of formula (I) the absolute stereochemical
configuration was not experimentally determined. In those cases the
stereochemically isomeric form which was first isolated is
designated as "A" and the second as "B", without further reference
to the actual stereochemical configuration.
[0084] A. Preparation of the Intermediates
EXAMPLE A.1
[0085] Triethylamine (9.2 ml) was added at room temperature to a
solution of indoline (20 g) in DCM (200 ml) and the mixture was
cooled till 5.degree. C. A solution of m-chlorocinnamoyl chloride
(40 g) in DCM (100 ml) was added dropwise and the mixture was
stirred at room temperature for 48 hours. Water was added, the
organic layer was decanted, washed with water, dried, filtered off
and evaporated. The residue was purified by column chromatography
over silica gel (eluent: cyclohexane/ethyl acetate 90/10), yielding
41 g (73%) of 1-[3-(3-chlorophenyl)-1-oxo-2-prop-
enyl]-2,3-dihydro-1H-indole (interm. 37).
[0086] In a similar way,
1-[3-(3-chlorophenyl)-1-oxo-2-propenyl]-1,2,3,4-t-
etrahydroquinoline (interm. 38) was synthesized.
EXAMPLE A.2
[0087] Intermediate 37 (40 g) and polyphosphoric acid (350 g) were
stirred and heated at 140.degree. C. for 16 hours. 4-Chlorobenzoic
acid (44 g) was added and the solution was stirred and heated at
140.degree. C. for 2 hours and 30 minutes. The mixture was cooled
till 80.degree. C., ice was added carefully and the mixture was
brought till room temperature. The precipitate was filtered off,
washed with water and basified with an aqueous ammonia solution.
The precipitate was taken up in DCM and filtered off. The organic
layer was dried, filtered and evaporated. The residue was purified
by column chromatography over silica gel (eluent:
CH.sub.2Cl.sub.2/CH.sub.3OH 99.5/0.5 to 99/1), yielding 12g (20%)
of
(.+-.)-8-(4-chlorobenzoyl)-6-(3-chlorophenyl)-1,2,5,6-tetrahydro-4H-pyrro-
lo[3,2,1-ij]quinolin-4-one (interm. 11).
[0088] In a similar way,
(.+-.)-9-(4-chlorobenzoyl)-7-(3-chlorophenyl--2,3-
,6,7-tetrahydro-1H,5H-benzo[ij]quinolizin-5-one (interm. 8) was
synthesized.
EXAMPLE A.3
[0089] A mixture of bromine (4.2 ml) in bromobenzene (80 ml) was
added dropwise at room temperature to a solution of intermediate 11
(34.2 g) in bromobenzene (300 ml). The mixture was stirred and
refluxed overnight. The mixture was cooled to room temperature and
basified with an aqueous ammonia solution. The solvent was
evaporated. The residue was partitioned between DCM and water. The
organic layer was separated, dried, filtered and the solvent was
evaporated. The residue was purified by column chromatography over
silica gel (eluent: DCM). Two fractions were collected, yielding 16
g of 8-(4-chlorobenzoyl)-6-(3-chlorophenyl)-1,2-di-
hydro4H-pyrrolo[3,2,1-ij]quinolin-4-one (intermediate 24) and 2.1 g
(6.2%) of
8-(4-chlorobenzoyl)-6-(3-chlorophenyl)-4H-pyrrolo[3,2,1-ij]quinolin-4--
one (interm. 25).
EXAMPLE A.4
[0090] A mixture of intermediate (9) (20.9 g), iodine (32.8 g) and
potassium acetate (19 g) in acetic acid (150 ml) was stirred at
130.degree. C. for 3 days. The mixture was poured out warm on ice
and NaHSO.sub.3 and extracted with DCM. The organic layer was
separated, dried, filtered and the solvent was evaporated till
dryness. The residue was purified by column chromatography over
silica gel (eluent: CH.sub.2Cl.sub.2/CH.sub.3OH 99/1 to 97/3. The
pure fractions were collected and the solvent was evaporated. The
residue was taken up in diethyl ether, filtered off and dried,
yielding 16.9 g (81%) of
8-(4-chlorobenzoyl)-1,2-dihydro-6-phenyl-4H-pyrrolo[3,2,1-ij]quinolin-4-o-
ne (interm. 21).
EXAMPLE A.5
[0091] a) A mixture of
(4-chlorophenyl)(3-methoxy-4-nitrophenyl)methanone (40.7 g),
1,2-ethanediol (31.2 ml) and 4-methylbenzene sulfonic acid (5.31 g)
in methylbenzene (320 ml) was stirred and refluxed using a
Dean-Stark apparatus. The mixture was washed with K.sub.2CO.sub.3
(10%) and extracted with DCM. The organic layer was separated,
dried, filtered and the solvent was evaporated. The residue was
crystallized from DIPE. The precipitate was filtered off and dried,
yielding 22.48 g (50.4%) of
2-(4-chlorophenyl)-2-(3-methoxy-4-nitrophenyl)-1,3-dioxolane
(interm. 39).
[0092] b) Intermediate (39) (22.48 g) and
3-chlorobenzenacetonitrile (15 ml) were added to a solution of
sodium hydroxide (11.25 g) in methanol (91 ml). The mixture was
stirred and refluxed for 24 hours. Ice water was added. The
precipitate was filtered off, washed with water and with ethanol
and dried. The residue was purified by column chromatography over
silica gel (eluent: CH.sub.2Cl.sub.2/cyclohexane 60/40). The pure
fractions were collected and the solvent was evaporated, yielding
8.5 g (27.3%) of
3-(3-chlorophenyl)-5-[2-(4-chlorophenyl)-1,3-dioxolan-2-yl]-7--
methoxy-2,1-benzisoxazole (interm. 40).
[0093] c) A mixture of intermediate (40) (14 g) in HCl conc. (3.5
ml) and THF (140 ml) was hydrogenated under a 2.4.times.10.sup.5 Pa
(2.4 bar) pressure for 6 hours with platinum on carbon (5%; 1.4 g)
as a catalyst in the presence of a 10% solution of thiophene in
methanol (0.35 ml). After uptake of adequate H.sub.2, the catalyst
was filtered through celite and the filtrate was evpaorated till
dryness. The residue was taken up in 2-propanone and DIPE. The
precipitate was filtered off and dried, yielding 11.8 g (84.3%) of
[2-amino-5-(4-chlorobenzoyl)-3-methoxyphenyl](-
3-chlorophenyl)methanone (interm. 41).
[0094] d) A mixture of intermediate (41) (11.7 g) and acetic
anhydride (28 ml) in toluene (150 ml) was stirred and refluxed for
24 hours. The solvent was evaporated till dryness. The product was
used without further purification, yielding 14.5 g of
N-acetyl-N-[2-(3-chlorobenzoyl)-4-(4-chl-
orobenzoyl)-6-methoxyphenyl]acetamide (interm. 42)
[0095] e) Potassium-tert-butoxide (13.5 g) was added portionwise to
a mixture of intermediate (42) (14.5 g) in dimethyl ether (150 ml).
The mixture was stirred at room temperature for 16 hours and then
hydrolized. The solvent was evaporated. Water was added. The
mixture was extracted with DCM and decanted. The organic layer was
dried, filtered and the solvent was evaporated till dryness,
yielding 11 g (86.6%) of
6-(4-chlorophenyl)-4-(3-chlorophenyl)-8-methoxy-2(1H)-quinolinone
(interm. 43).
[0096] f) A boron tribromide solution in DCM (1M; 95 ml) was added
dropwise at 0.degree. C. to a mixture of intermediate (43) (10 g)
in DCM (100 ml). The mixture was stirred at room temperature
overnight, then hydrolized, alkalized with K.sub.2CO.sub.3 (10%)
and extracted with CH.sub.2Cl.sub.2/CH.sub.3OH 90/10. The organic
layer was separated, dried, filtered and the solvent was evaporated
till dryness. The product was used without further purification,
yielding 9.6 g of
6-(4-chlorophenyl)-4-(3-chlorophenyl)-8-hydroxy-2(1H)-quinolinone
(interm. 44)
[0097] g) A mixture of intermediate (44) (15 g), 1,2-dibromoethane
(12.6 ml), potassium carbonate (20.2 g) and
tricaprylylmethylammonium chloride (Aliquat 336) (1.6 ml) in ACN
(120 ml) and DCM (180 ml) was stirred at 50.degree. C. for 24 hours
and then cooled to room temperature. Water was added. The mixture
was decanted and extracted with DCM. The organic layer was
separated, dried, filtered and the solvent was evaporated till
dryness. The residue was purified by column chromatography over
silica gel (eluent: CH.sub.2Cl.sub.2/EtOAc 95/5). Two pure
fractions were collected and their solvents were evaporated,
yielding 4.5 g (28.6%) of
9-(4-chlorophenyl)-7-(3-chlorophenyl)-2,3-dihydro-5H-pyrido[1,2,3-de]-1,4-
-benzoxazin-5-one (interm. 45).
[0098] h) Sodium borohydride (NaBH.sub.4) (0.21 g) was added at
5.degree. C. to a mixture of intermediate (45) (2.5 g) in methanol
(30 ml) and THF (30 ml). The mixture was stirred at 5.degree. C.
for 30 minutes, then hydrolized, extracted with DCM and decanted.
The organic layer was dried, filtered and the solvent was
evaporated till dryness, yielding 2.3 g of
(.+-.)-7-(3-chlorophenyl)-9-[(4-chlorophenyl)hydroxymethyl]-2,3-dihydro-5-
H-pyrido[1,2,3-de]-1,4-benzoxazin-5-one (interm. 46).
[0099] i) A mixture of intermediate (46) (2.3 g) in thionylchloride
(30 ml) was stirred at room temperature for 16 hours. The solvent
was evaporated till dryness. The product was used without further
purification, yielding 2.6 g of
(.+-.)-9-[chloro(4-chlorophenyl)methyl]-7-
-(3-chlorophenyl)-2,3-dihydro-5H-pyrido[1,2,3-de]-1,4-benzoxazin-5-one
(intermediate 47).
[0100] In a similar way,
(.+-.)-8-[chloro(4-chlorophenyl)methyl]-6-(3-chlo-
rophenyl)-2H,4H-oxazolo[5,4,3-ij]quinolin-4-one (interm. 48) was
also prepared.
EXAMPLE A.6
[0101] Intermediate (37) (23 g) and polyphosphoric acid (PPA) (120
g) were stirred at 140.degree. C. for 24 hours. The mixture was
poured out into ice water, filtered, washed with water, stirred in
NH.sub.3 (aq.), washed with water and extracted with DCM. The
organic layer was separated, dried, filtered and the solvent was
evaporated. The residue was purified by column chromatography over
silica gel (eluent: DCM). The pure fractions were collected and the
solvent was evaporated. Part of this fraction was crystallized from
diethyl ether/2-propanone. The precipitate was filtered off and
dried, yielding 0.6 g of (.+-.)-6-(3-chlorophenyl)-1-
,2,5,6-tetrahydro-4H-pyrrolo[3,2,1-ij]quinolin-4-one (interm.
3).
EXAMPLE A.7
[0102] a) Pyridine (25 ml) was added to a mixture of
3,4-dihydro-2H-1,4-benzoxazine (17.8 g) in DCM (200 ml). The
mixture was cooled on an ice bath and poured out into a mixture of
m-chloro-cinnamoyl chloride (33 g) in DCM (100 ml). The mixture was
stirred at room temperature overnight. Water was added and the
mixture was decanted. The organic layer was dried, filtered and the
solvent was evaporated till dryness, to give a residue which was
purified by column chromatography over silica gel (eluent:
cyclohexane/ethyl acetate 80/20) and recrystallisation from
ACN/diethyl ether, yielding 26 g (65.8%) of
4-[3-(3-chlorophenyl)-1-oxo-2-propen-1-yl]-2,3-dihydro-4H-1,4-benzoxazine
(interm. 51).
[0103] b) AlCl.sub.3 (7.2 g) was added to a mixture of intermediate
(51) (5 g) in chlorobenzene (50 ml). The mixture was stirred and
refluxed at 80.degree. C. for 2 hours, then poured out on ice and
extracted with DCM. The organic layer was separated, dried,
filtered and the solvent was evaporated. The residue was taken up
in DCM, filtered off, washed with CH.sub.2Cl.sub.2/diethyl ether
and dried, to givea residue which was purified by column
chromatography over silica gel (eluent: CH.sub.2Cl.sub.2/CH.sub.3OH
99/1), yielding 3.4 g (62%) of
(.+-.)-7-(3-chlorophenyl)-2,3,6,7-tetrahydro-5H-pyrido[1,2,3-de]-1,4-benz-
oxazin-5-one (interm. 7).
EXAMPLE A.8
[0104] a) Butyllithium (1.6 M in hexanes, 22.4 ml) was added at
-70.degree. C. under nitrogen flow to a mixture of
1-methylimidazole (2.94 g) in THF (50 ml). The mixture was stirred
at -70.degree. C. for 30 minutes. Triethylsilyl chloride (6 ml) was
added. The mixture was brought to room temperature and cooled to
-70.degree. C. Butyllithium (1.6 M in hexanes, 22.4 ml) was added.
The mixture was stirred at -70.degree. C. for 1 hour, then brought
to -15.degree. C. and cooled to -70.degree. C. Intermediate 12 (3.8
g) was added portionwise. The mixture was brought to -10.degree. C.
Water was added and the mixture was extracted with ethyl acetate
and a small amount of methanol. The organic layer was separated,
dried, filtered and the solvent was evaporated. The residue was
purified by column chromatography over silica gel (eluent:
CH.sub.2Cl.sub.2/CH.sub- .3OH/NH.sub.4OH 95/5/0.2). The pure
fractions were collected and the solvent was evaporated, yielding 4
g (88%) of (.+-.)-4-(3-chlorophenyl)-6-
-[(4-chlorophenyl)hydroxy(1-methyl-1H-imidazol-5-yl)methyl]-8-methoxy-2(1H-
)-quinolinone (interm. 49).
[0105] b) A boron tribromide solution in DCM (1M; 27.6 ml) was
added dropwise at 10.degree. C. to a solution of intermediate (49)
(2.8 g) in DCM (30 ml). The mixture was stirred at room temperature
for 5 hours. Water was added slowly. The mixture was stirred at
room temperature overnight. The precipitate was filtered off,
washed with water and dried, yielding 2.9 g (100%) of
(.+-.)-4-(3-chlorophenyl)-6-[(4-chlorophenyl)hyd-
roxy(1-methyl-1H-imidazol-5-yl)methyl]-8-hydroxy-2(1H)-quinolinone
(interm. 50).
EXAMPLE A.9
[0106] Sodium borohydride (0.51 g) was added portionwise at room
temperature to a solution of intermediate (23) (2.9 g) in methanol
(20 ml) and THF (10 ml) and the mixture was stirred at room
temperature for 1 hour. The mixture was poured into water and
evaporated. Methanol was added, the mixture was extracted with DCM
and decanted. The organic layer was dried, filtered and evaporated,
yielding 2.9 g (100%) of
(.+-.)-7-(3-chlorophenyl)-9-[hydroxy(4-chlorophenyl)methyl]-2,3-dihydro-1-
H,5H-benzo[ij]-quinolizin-5-one (interm. 52).
[0107] In a similar way,
(.+-.)-7-(3-chlorophenyl)-9-[hydroxy(4-chlorophen-
yl)methyl]-2,3,6,7-tetrahydro-1H,5H-benzo[ij]quinolizin-5-one
(interm. 53) was synthesized.
EXAMPLE A. 10
[0108] Methanesulfonyl chloride (1.6 ml) was added dropwise at room
temperature to a solution of intermediate (52) (2.6 g) and
triethylamine (4.1 ml) in DCM (30 ml) and the mixture was stirred
at room temperature for 2 hours. The mixture was poured into water
and decanted. The organic layer was dried, filtered and evaporated,
yielding 3.4 g of
(.+-.)-7-(3-chlorophenyl)-9-[hydroxy(4-chlorophenyl)methyl]-2,3,6,7-tetra-
hydro-1H,5H-benzo[ij]quinolizin-5-one (interm. 54).
EXAMPLE A.11
[0109] a) 1,1'-Carbonyldiimidazole (41 g) was added portionwise at
room temperature to a mixture of 2-amino-5-bromo-3-nitro-benzoic
acid (55 g) in DCM (700 ml). The mixture was stirred at room
temperature for 1 hour. N-methoxy-methanamine hydrochloride (24.6
g) was added. The mixture was stirred at room temperature overnight
and hydrolized with water. The precipitate was filtered off and the
filtrate was decanted. The organic layer was dried, filtered and
the solvent was evaporated till dryness. The residue was purified
by column chromatography over silica gel (eluent:
CH.sub.2Cl.sub.2/ethyl acetate 98/2). The pure fractions were
collected and the solvent was evaporated. The precipitate was taken
up in HCl 3N (250 ml). The mixture was stirred at room temperature
for 4 hours. The precipitate was filtered off, washed with water
and dried, yielding 23 g of
2-amino-5-bromo-N-methoxy-N-methyl-3-nitrobenzamide (interm. 55,
mp. 129.degree. C.)
[0110] b) A mixture of 1-bromo-3-chlorophenyl (37.3 ml) in THF (300
ml) was added dropwise to a mixture of magnesium (7.7 g) in a small
amount of THF, while the temperature was kept at 50.degree.
C.-60.degree. C. The mixture was stirred at room temperature for 1
hour and cooled to 5.degree. C. A mixture of intermediate (55)
(30.7 g) in THF (300 ml) was added dropwise. The mixture was
stirred at 5.degree. C. for 15 minutes, hydrolized, extracted with
ethyl acetate, filtered over celite and decanted. The organic layer
was dried, filtered and the solvent was evaporated till dryness.
The residue was purified by column chromatography over silica gel
(eluent: CH.sub.2Cl.sub.2/cyclohexane 50/50). The pure fractions
were collected and the solvent was evaporated, yielding 17.5 g
(46.4%) of (2-amino-5-bromo-3-nitrophenyl)(3-chlorophenyl-
)methanone (interm. 56, mp. 134.degree. C.).
[0111] c) TiCl.sub.3 (15% in H.sub.2O, 400 ml) was added at room
temperature to a solution of intermediate (56) (16 g) in THF (230
ml). The mixture was stirred at room temperature overnight. Water
was added and the mixture was extracted twice with DCM. The
combined organic layer was washed with K.sub.2CO.sub.3 10%, dried,
filtered and the solvent was evaporated, yielding 18 g of
(2,3-diamino-5-bromophenyl)(3-chlorophenyl)m- ethanone (interm.
57)
[0112] d) A mixture of interm. (57) (18 g) and acetic acid
anhydride (19 ml) in toluene (400 ml) was stirred and refluxed for
4 hours and then allowed to cool to room temperature. The
precipitate was filtered off, washed with DIPE and dried, yielding
13.2 g (90%) of
N,N'-[5-bromo-3-(3-chlorobenzoyl)-1,2-phenylene]diacetamide
(interm. 58).
[0113] e) Potassium tert-butoxide (18 g) was added at room
temperature to a mixture of interm. (58) (13.2 g) in DME (140 ml).
The mixture was stirred at room temperature overnight. Water was
added and the mixture was neutralized with HCl 3N. The precipitate
was filtered off, washed with water and with DIPE and dried,
yielding 10.75 g (86%) of
N-[6-bromo-4-(3-chlorophenyl)-1,2-dihydro-2-oxo-8-quinolinyl]acetamide
(interm. 59).
[0114] f) A mixture of interm. (59) (10.75 g), methyl iodide (3.57
ml) and Ag.sub.2CO.sub.3 (16.93 g) in DMF (150 ml) was stirred at
80.degree. C. under N.sub.2 flow for 90 minutes. The mixture was
allowed to cool to room temperature. Water was added. The mixture
was filtered over celite, washed with water and extracted with DCM.
The organic layer was separated, dried, filtered and the solvent
was evaporated, yielding 10.9 g (98%) of
N-[6-bromo-4-(3-chlorophenyl)-2-methoxy-8-quinolinyl]acetamide
(interm. 60).
[0115] g) Butyllithium (1.6 M in hexanes, 18.5 ml) was added
dropwise at -70.degree. C. under N.sub.2 flow to a mixture of
interm. (60) (5 g) in THF (70 ml). The mixture was stirred at
-70.degree. C. for 30 minutes, brought to -40.degree. C. and cooled
again to -70.degree. C. (4-Chlorophenyl)
(1-methyl-1H-imidazol-5-yl)methanone (6.5 g) was added. The mixture
was allowed to warm to room temperature and then hydrolized. Ethyl
acetate was added. The organic layer was separated, dried, filtered
and the solvent was evaporated till dryness. The residue was
purified by column chromatography over silica gel (eluent:
CH.sub.2Cl.sub.2/CH.sub.3O- H/NH.sub.4OH 95/5/0.1). The pure
fraction was collected, evaporated, recrystallized from
2-propanone, ACN and DIPE, yielding 1.3 g (32.5%) of
(.+-.)--N--[4-(3-chlorophenyl)-6-[(4-chlorophenyl)hydroxy(1-methyl-1H-imi-
dazol-5-yl)methyl]-2-methoxy-8-quinolinyl]acetamide (interm. 61,
mp. 143.degree. C.).
[0116] h) A mixture of interm. (61) (3 g) in HBr (48% in H.sub.2O,
45 ml) and 1,4-dioxane (40 ml) was stirred at 80.degree. C. for 3
hours. The mixture was cooled to room temperature, poured out on
ice, saturated with K.sub.2CO.sub.3 solid and extracted with ethyl
acetate. The organic layer was separated, dried, filtered,
evaporated and purified by column chromatography over silica gel
(eluent: CH.sub.2Cl.sub.2/CH.sub.3OH/NH.su- b.4OH 95/5/0.5). The
pure fractions were collected and the solvent was evaporated. The
residue was taken up in CH.sub.3OH and DIPE. The precipitate was
filtered off and dried, yielding 0.4 g (55%) of
(.+-.)-8-amino-4-(3-chlorophenyl)-6-[(4-chlorophenyl)hydroxy(1-methyl-1H--
imidazol-5-yl)methyl]-2(1H)-quinolinone(interm. 62).
[0117] Tables I-1 to I-2 list intermediates prepared according to
one of the above Examples.
5TABLE I-1 16 Physical Intm. No. Ex. No. -A- R.sup.1 R.sup.2 data 1
A.6 --(CH.sub.2).sub.3-- 3-Cl H -- 2 A.6 --(CH.sub.2).sub.2-- 3-Br
H -- 3 A.6 --(CH.sub.2).sub.2-- 3-Cl H mp. 138.degree. C. 4 A.6
--(CH.sub.2).sub.2-- 4-Cl H -- 5 A.6 --(CH.sub.2).sub.2-- 3-Cl 4-Cl
-- 6 A.6 --(CH.sub.2).sub.2-- 3-CH.sub.3 H -- 7 A.7
--(CH.sub.2*).sub.2--O-- 3-Cl H -- *the CH.sub.2 moiety is linked
to the nitrogen atom of the 2-quinolinone moiety
[0118]
6TABLE 1-2 17 Phys- Intm. Ex. ical No. No. 18 -A- R.sup.1 R.sup.2
R.sup.3 data 8 A.2 single --(CH.sub.2).sub.3-- 3-Cl H 4-Cl mp.
176.degree. C. 9 A.2 single --(CH.sub.2).sub.2-- H H 4-Cl -- 10 A.2
single --(CH.sub.2).sub.2-- 3-Br H 4-Cl -- 11 A.2 single
--(CH.sub.2).sub.2-- 3-Cl H 4-Cl mp. 140.degree. C. 12 A.2 single
--(CH.sub.2).sub.2-- 3-Cl H 4-F -- 13 A.2 single
--(CH.sub.2).sub.2-- 3-Cl H 3-Cl 14 A.2 single --(CH.sub.2).sub.2--
3-Cl 4-Cl 4-Cl -- 15 A.2 single --(CH.sub.2).sub.2-- 3-Cl H H -- 16
A.2 single --(CH.sub.2).sub.2-- 3-Cl H 4-CH.sub.3 -- 17 A.2 single
--(CH.sub.2).sub.2-- 3-Cl H 2-Cl -- 18 A.2 single
--(CH.sub.2).sub.2-- 3-Cl H 4-OCH.sub.3 -- 19 A.2 single
--(CH.sub.2).sub.2-- 4-Cl H 4-Cl -- 20 A.2 single
--(CH.sub.2).sub.2-- 3- H 4-Cl -- CH.sub.3 21 A.4 dou-
--(CH.sub.2).sub.2-- H H 4-Cl -- ble 22 A.4 dou-
--(CH.sub.2).sub.2-- 3-Br H 4-Cl -- ble 23 A.4 dou-
--(CH.sub.2).sub.3-- 3-Cl H 4-Cl mp. ble 194.degree. C. 24 A.3 dou-
--(CH.sub.2).sub.2-- 3-Cl H 4-Cl mp. ble 191.degree. C. 25 A.3 dou-
--CH.dbd.CH-- 3-Cl H 4-Cl -- ble 26 A.5 dou-
--(CH.sub.2*).sub.2--O-- 3-Cl H 4-Cl mp. ble 135.degree. C. 27 A.5
dou- --CH.sub.2*--O-- 3-Cl H 4-Cl mp. ble 154.degree. C. 28 A.4
dou- --(CH.sub.2).sub.2-- 3-Cl H 4-F -- ble 29 A.4 dou-
--(CH.sub.2).sub.2-- 3-Cl H 3-Cl -- ble 30 A.4 dou-
--(CH.sub.2).sub.2-- 3-Cl 4-Cl 4-Cl -- ble 31 A.4 dou-
--(CH.sub.2).sub.2-- 3-Cl H H -- ble 32 A.4 dou-
--(CH.sub.2).sub.2-- 3-Cl H 4-CH.sub.3 -- ble 33 A.4 dou-
--(CH.sub.2).sub.2-- 3-Cl H 2-Cl -- ble 34 A.4 dou-
--(CH.sub.2).sub.2-- 3-Cl H 4-OCH.sub.3 -- ble 35 A.4 dou-
--(CH.sub.2).sub.2-- 4-Cl H 4-Cl -- ble 36 A.4 dou-
--(CH.sub.2).sub.2-- 3- H 4-Cl -- ble CH.sub.3 *the CH.sub.2 moiety
is linked to the nitrogen atom of the 2-quinolinone moiety
[0119] B. Preparation of the Final Compounds
EXAMPLE B.1
[0120] A solution of 1-methylimidazole (4.55 ml) in THF (200 ml)
was cooled to -70.degree. C. Butyllithium (1.6 M in hexanes, 35.9
ml) was added and the mixture was stirred at -70.degree. C. for 30
minutes. Triethylsilyl chloride (10.4 ml) was added and the mixture
was allowed to warm to room temperature slowly. The mixture was
cooled to -70.degree. C. and butyllithium (1.6 M in hexanes 35.9
ml) was added dropwise. The mixture was stirred at -70.degree. C.
for 1 hour and was then allowed to warm to -15.degree. C. The bath
was removed and the mixture was cooled to -70.degree. C.
Intermediate (24) (20 g) was added and the mixture was stirred at
-70.degree. C. for 30 minutes. The mixture was hydrolyzed and
extracted with ethyl acetate. The organic layer was separated,
dried, filtered and the solvent was evaporated. The residue was
purified by column chromatograph (eluent:
CH.sub.2Cl.sub.2/CH.sub.3OH/NH.sub.4OH 97/3/0.1), yielding 24 g of
(.+-.)-6-(3-chlorophenyl)-8-[(4-chlorophenyl)-
hydroxy(1-methyl-1H-imidazol-5-yl)methyl]-1,2-dihydro-4H-pyrrolo[3,2,1-ij]-
quinolin-4-one (comp. 5, mp. 213.6.degree. C.).
EXAMPLE B.2
[0121] A mixture of compound 1 (2.5 g) in formamide (10 ml) and
acetic acid (20 ml) was stirred at 160.degree. C. for 4 hours. The
mixture was poured out on ice, basified with an aqueous ammonia
solution and extracted with DCM. The organic layer was separated,
dried, filtered and the solvent was evaporated. The residue was
purified by column chromatography over silica gel (eluent:
CH.sub.2Cl.sub.2/CH.sub.3OH/NH.su- b.4OH 97/3/0.1). The pure
fractions were collected and the solvent was evaporated. The
residue was taken up in 2-propanone/DIPE. The precipitate was
filtered off and dried, yielding 1 g (41%) of
(.+-.)-6-(3-chloropheny-
l)-8-[(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-1,2-dihydro-4H-py-
rrolo[3,2,1-ij]quinolin-4-one monohydrate (comp. 6, mp.
147.0.degree. C.).
EXAMPLE B.3
[0122] A mixture of compound 5 (2 g) in thionyl chloride (8 ml) was
stirred at room temperature overnight. The solvent was evaporated
till dryness. The product was used without further purification,
yielding 2.07 g (100%) of
(.+-.)-8-[chloro(4-chlorophenyl)-(1-methyl-1H-imidazol-5-yl)m-
ethyl]-6-(3-chlorophenyl)-1,2-dihydro4H-pyrrolo[3,2,1-ij]quinolin-4-one
monohydrochloride (comp. 7).
EXAMPLE B.4
[0123] A mixture of compound 7 (2.07 g) in THF (15 ml) was poured
out into an aqueous ammonia solution (40 ml) at room temperature.
The mixture was stirred at room temperature for 4 hours, then
extracted with DCM and decanted. The organic layer was dried,
filtered and the solvent was evaporated. The residue was purified
by column chromatography over silica gel (eluent:
toluene/2-propanol/NH.sub.4OH 50/50/1). The pure fractions were
collected and the solvent was evaporated. The residue was
recrystallized from CH.sub.2Cl.sub.2/diethyl ether. The precipitate
was filtered off and dried, yielding 0.65 g
(.+-.)-8-[amino(4-chlorophenyl)(1-
-methyl-1H-imidazol-5-yl)methyl]-6-(3-chlorophenyl)-1,2-dihydro-4H-pyrrolo-
[3,2,1-ij]quinolin-4-one (comp. 8).
EXAMPLE B.5
[0124] Compound 8 (12.4 g) was separated and purified by chiral
column chromatography over Chiracel OD (eluent: 100% CH.sub.3OH).
Two pure fraction groups were collected. The solvent of the first
fraction group was evaporated. The residue was crystallized from
2-propanol (200 ml) and DIPE (200 ml). The precipitate was filtered
off and dried, yielding 4.4 g of
(A)-8-[amino(4-chlorophenyl)(1-methyl-1H-imidazol-5-yl)methyl]-6-(3-ch-
lorophenyl)-1,2-dihydro-4H-pyrrolo[3,2,1-ij]quinolin-4-on (compound
9; [.alpha.].sub.D.sup.20=-27.94.degree. (c=9.1 mg/ml in
methanol)). The solvent of the second fraction group was
evaporated. The residue was crystallized from 2-propanol (250 ml)
and DIPE (350 ml). The precipitate was filtered off and dried,
yielding: 4.1 g of (B)-8-[amino(4-chloropheny-
l)(1-methyl-1H-imidazol-5-yl)methyl]-6-(3-chlorophenyl)-1,2-dihydro-4H-pyr-
rolo[3,2,1-ij]quinolin-4-on (compound 10;
[.alpha.].sub.D.sup.20=+28.21.de- gree. (c=9 mg/ml in
methanol)).
EXAMPLE B.6
[0125] A mixture of intermediate (50) (2.7 g), dibromomethane (3
ml) and potassium carbonate (2.8 g) in DMF (90 ml) was stirred at
80.degree. C. for 3 hours. Water was added. The mixture was
filtered over celite, washed with water and extracted with DCM. The
organic layer was separated, dried, filtered and the solvent was
evaporated. The residue was purified by column chromatography over
silica gel (eluent: CH.sub.2Cl.sub.2/CH.sub.3OH/NH.sub.4OH
96/4/0.2). The pure fractions were collected and the solvent was
evaporated. The residue was crystallized from 2-propanone and
diethyl ether. The precipitate was filtered off and dried, yielding
0.86g (31%) of (.+-.)-6-(3-chlorophenyl)-8-[(4-chlorophen-
yl)hydroxy(1-methyl-1H-imidazol-5-yl)methyl]-2H,4H-oxazolo[5,4,3-ij]quinol-
in-4-one (comp. 15).
EXAMPLE B.7
[0126] A mixture of compound 6 (1.2 g) and phosphorus sulfide (2.4
g) in pyridine (30 ml) was stirred and refluxed for 6 hours and
then poured out into water. The precipitate was filtered off,
rinced abundantly with water, taken up in DCM, dried, filtered and
the solvent was evaporated till dryness. The residue was purified
by column chromatography over silica gel (eluent:
CH.sub.2Cl.sub.2/CH.sub.3OH/NH.sub.4OH 98/2/0.1). The pure
fractions were collected and the solvent was evaporated. The
residue was crystallized from ACN and DIPE. The precipitate was
filtered off and dried, yielding 0.36 g (29.2%) of
(.+-.)-6-(3-chlorophenyl)-8-[(4-chlorop-
henyl)(1-methyl-1H-imidazol-5-yl)methyl]-1,2-dihydro-4H-pyrrolo[3,2,1-ij]q-
uinoline-4-thione (comp. 27).
EXAMPLE B.8
[0127] A mixture of interm. (62) (1.8 g) and ethyl acetimidate (0.9
g) in methanol (40 ml) was stirred and refluxed for 4 hours. The
solvent was evaporated till dryness. The residue was taken up in
DCM and K.sub.2CO.sub.3 (10% in H.sub.2O). The organic layer was
separated, dried, filtered and the solvent was evaporated. The
residue was purified by column chromatography over silica gel
(eluent: CH.sub.2Cl.sub.2/CH.sub- .3OH/NH.sub.4OH 95/5/0.5). The
pure fractions were collected and the solvent was evaporated. The
residue was crystallized from DIPE. The precipitate was filtered
off and dried, yielding 0.4 g (21.2%) of
(.+-.)-6-(3-chlorophenyl)-8-[(4-chlorophenyl)hydroxy(1-methyl-1H-imidazol-
-5-yl)methyl]-2-methyl-4H-imidazo[4,5,1-ij]quinolin-4-one (comp.
30, mp. 170.degree. C.). In a similar way,
(.+-.)-6-(3-chlorophenyl)-8-[(4-chloro-
phenyl)hydroxy(1-methyl-1H-imidazol-5-yl)methyl]-2-phenyl-4H-imidazo[4,5,1-
-ij]quinolin-4-one (comp. 31) was also prepared.
EXAMPLE B.9
[0128] A mixture of interm. (62) (2.1 g) and
1,1'-carbonyldiimidazole (4.1) in THF (60 ml) was stirred and
refluxed for 3 hours. The mixture was poured out into water and
extracted with DCM. The organic layer was separated, washed with
water, dried, filtered and the solvent was evaporated till dryness.
The residue was purified by column chromatography over silica gel
(eluent: CH.sub.2Cl.sub.2/CH.sub.3OH/NH.su- b.4OH 90/10/0.5). The
pure fractions were collected and the solvent was evaporated. The
residue was crystallized from CH.sub.3OH and DIPE. The precipitate
was filtered off and dried, yielding 0.7 g (31.8%) of
(.+-.)-6-(3-chlorophenyl)-8-[(4-chlorophenyl)hydroxy(1-methyl-1H-imidzol--
5-yl)methyl]-4H-imidazo[4,5,1-ij]quinoline-2,4(1H)-dione(comp. 34,
mp. 256.degree. C.).
EXAMPLE B.10
[0129] A mixture of interm. (62) (2.1 g) in water (21 ml) and
sulfuric acid (36 N, 42 ml) was cooled to 5.degree. C. on an ice
bath. NaNO.sub.2 (3.6 ml; solution 80 g/100 ml) was added dropwise
while the temperature was kept at 5.degree. C. The mixture was
stirred for 1 hour on an ice bath, poured out into ice water,
alkalized with a concentrated NH.sub.4OH solution and extracted
with DCM. The organic layer was separated, dried, filtered and the
solvent was evaporated. The residue was purified by column
chromatography over silica gel (eluent: CH.sub.2Cl.sub.2/CH.sub.3O-
H/NH.sub.4OH 97.5/2.5/0.1). The pure fractions were collected and
the solvent was evaporated. The residue was crystallized from
2-propanone and DIPE. The precipitate was filtered off and dried,
yielding 0.35 g (16.3%) of
(.+-.)-6-(3-chlorophenyl)-8-[(4-chlorophenyl)hydroxy(1-methyl-1H-imida-
zol-5-yl)methyl]-4H-1,2,3-triazolo[4,5,1-ij]quinolin-4-one (comp.
35, mp. 226.degree. C.).
EXAMPLE B.11
[0130] A mixture of intermediate (54) (3.4 g) and imidazole (2.01
g) in ACN (40 ml) was stirred and refluxed for 3 hours. The mixture
was evaporated and the residue was taken up in DCM. The organic
layer was washed with water, dried, filtered off and evaporated.
The residue was purified by column chromatography over silica gel
(eluent: CH.sub.2Cl.sub.2/CH.sub.3OH/NH.sub.4OH 97/3/0.1). The pure
fractions were collected and evaporated. Crystallisation from ethyl
acetate and DIPE yielded 1.9 g (65%)
(.+-.)-7-(3-chlorophenyl)-9-[(4-chlorophenyl)-1H-imid-
azol-1-ylmethyl]-2,3-dihydro-1H,5H-benzo[ij]quinolizin-5-one (comp.
36, mp. 195.2.degree. C.).
EXAMPLE B.12
[0131] 1,1'-Carbonyldiimidazole (4 g) was added at room temperature
to a solution of intermediate (53) (5.4 g) in THF (70 ml) and the
mixture was stirred at room temperature for 16 hours. Water was
added and the mixture was extracted with DCM. The organic layer was
dried, filtered and evaporated. The residue was purified by column
chromatography over silica gel (eluent:
CH.sub.2Cl.sub.2/CH.sub.3OH/NH.sub.4OH 97.5/2.5/0.1). The pure
fractions were collected and evaporated. The residue was purified
further by column chromatography over silica gel (eluent:
cyclohexane/2-propanol/NH.sub.4OH 80/20/0.1). The pure fractions
were collected and evaporated. The residue was taken up in diethyl
ether and filtered off, yielding 1.3 g (22%) of
(.+-.)-7-(3-chlorophenyl)-9-[(4-chl-
orophenyl)-1H-imidazol-1-ylmethyl]-2,3,6,7-tetrahydro-1H,5H-benzo[ij]quino-
lizin-5-one (comp. 37, mp. 93.6.degree. C.).
EXAMPLE B.13
[0132] A mixture of intermediate (48) (2.3 g) and imidazole (1.8 g)
in ACN (50 ml) was stirred and refluxed for 4 hours. The solvent
was evaporated till dryness. The residue was taken up in DCM,
washed with water and decanted. The organic layer was dried,
filtered and the solvent was evaporated till dryness. The residue
was purified by column chromatography over silica gel (eluent:
CH.sub.2Cl.sub.2/CH.sub.3OH/NH.su- b.4OH 98/2/0.1). The pure
fractions were collected and the solvent was evaporated. The
residue was crystallized from ACN and DIPE. The precipitate was
filtered off and dried, yielding 1.3 g of
(.+-.)-6-(3-chlorophenyl)-8-[(4-chlorophenyl)-1H-imidazol-1-ylmethyl]-2H,-
4H-oxazolo[5,4,3-ij]quinolin-4-one (comp. 52).
EXAMPLE B.14
[0133] Phosphorus sulfide (6 g) was added to a mixture of compound
38 (3 g) in pyridine (40 ml). The mixture was stirred and refluxed
for 6 hours. Ice water was added. The precipitate was filtered off,
washed with water and taken up in DCM. The organic layer was
separated, dried, filtered and the solvent was evaporated till
dryness. The residue was purified by column chromatography over
silica gel (eluent: CH.sub.2Cl.sub.2/CH.sub.3O- H 98.5/1.5). The
pure fractions were collected and the solvent was evaporated. The
residue was crystallized from ACN and DIPE. The precipitate was
filtered off and dried, yielding 1.1 g of
(.+-.)-6-(3-chlorophenyl)-8-[(4-chlorophenyl)-1H-imidazol-1-ylmethyl]-1,2-
-dihydro-4H-pyrrolo[3,2,1-ij]quinoline-4-thione (comp. 50).
[0134] Tables F-1 and F-3 list the compounds that were prepared
according to one of the above Examples.
7TABLE F-1 19 Co. Ex. No. No. X -A- R.sup.1 R.sup.2 R.sup.3 R.sup.4
R.sup.6 Physical data 1 B.1 O --(CH.sub.2).sub.2-- H H 4-Cl H OH
mp. 240.degree. C. 2 B.3 O --(CH.sub.2).sub.2-- H H 4-Cl H Cl (2) 3
B.4 O --(CH.sub.2).sub.2-- H H 4-Cl H NH.sub.2 mp. 218.degree. C. 4
B.1 O --(CH.sub.2).sub.2-- 3-Br H 4-Cl H OH mp. 210.degree. C. 5
B.1 O --(CH.sub.2).sub.2-- 3-Cl H 4-Cl H OH mp. 213.6.degree. C. 6
B.2 O --(CH.sub.2).sub.2-- 3-Cl H 4-Cl H H mp. 147.0.degree. C.;
(1) 7 B.3 O --(CH.sub.2).sub.2-- 3-Cl H 4-Cl H Cl (2) 8 B.4 O
--(CH.sub.2).sub.2-- 3-Cl H 4-Cl H NH.sub.2 mp. 165.degree. C. 9
B.5 O --(CH.sub.2).sub.2-- 3-Cl H 4-Cl H NH.sub.2 (A);
[.alpha.].sup.20.sub.D = -27.94.degree. 10 B.5 O
--(CH.sub.2).sub.2-- 3-Cl H 4-Cl H NH.sub.2 (B);
[.alpha.].sup.20.sub.D = +28.21.degree. 11 B.1 O
--(CH.sub.2).sub.3-- 3-Cl H 4-Cl H OH mp. 210.degree. C.(dec.) 12
B.1 O --(CH.sub.2).sub.3-- 3-Cl H 4-Cl H OH mp. 232.degree. C.; (2)
13 B.3 O --(CH.sub.2).sub.3-- 3-Cl H 4-Cl H Cl (2) 14 B.4 O
--(CH.sub.2).sub.3-- 3-Cl H 4-Cl H NH.sub.2 mp. 190.degree. C. 15
B.6 O --CH.sub.2*--O-- 3-Cl H 4-Cl H OH mp. 196.degree. C. 16 B.3 O
--(CH.sub.2).sub.2--O-- 4-Cl H 4-Cl H Cl (2) 17 B.1 O
--(CH.sub.2*).sub.2--O-- 3-Cl H 4-Cl H OH mp. 252.degree. C. 18 B.4
O --(CH.sub.2*).sub.2--O-- 3-Cl H 4-Cl H NH.sub.2 mp. 210.degree.
C. 19 B.1 O --(CH.sub.2).sub.2-- 3-Cl H 4-F H OH mp. 224.degree. C.
20 B.3 O --(CH.sub.2).sub.2-- 3-Cl H 4-F H Cl (2) 21 B.4 O
--(CH.sub.2).sub.2-- 3-Cl H 4-F H NH.sub.2 mp. 235.degree. C. 22
B.1 O --(CH.sub.2).sub.2-- 3-Cl H 3-Cl H OH mp. 220.degree. C. 23
B.1 O --(CH.sub.2).sub.2-- 3-Cl H H H OH mp. 260.degree. C. 24 B.1
O --(CH.sub.2).sub.2-- 3-Cl H 4-CH.sub.3 H OH mp. 245.degree. C. 25
B.1 O --(CH.sub.2).sub.2-- 3-Cl H 2-Cl H OH mp. 210.degree. C.; (1)
26 B.1 O --(CH.sub.2).sub.2-- 3-CH.sub.3 H 4-Cl H OH mp.
242.degree. C. 27 B.7 S --(CH.sub.2).sub.2-- 3-Cl H 4-Cl H H mp.
138.degree. C. 30 B.8 O --CH(CH.sub.3).dbd.N-- 3-Cl H 4-Cl H OH mp.
170.degree. C. 31 B.6 O --CH(C.sub.6H.sub.5).dbd.N-- 3-Cl H 4-Cl H
OH mp. 176.degree. C. 32 B.3 O --CH(C.sub.6H.sub.5).dbd.N-- 3-Cl H
4-Cl H Cl (1) 33 B.4 O --CH(C.sub.6H.sub.5).dbd.N-- 3-Cl H 4-Cl H
NH.sub.2 mp. 215.degree. C. 34 B.9 O --CO--NH-- 3-Cl H 4-Cl H OH
mp. 256.degree. C. 35 B.10 O --N.dbd.N-- 3-Cl H 4-Cl H OH mp.
226.degree. C. *the CH.sub.2 moiety is linked to the nitrogen atom
of the 2-quinolinone moiety (1): hydrate (1:1) (2): hydrochloride
(1:1)
[0135]
8TABLE F-2 20 Phys- Co. Ex. ical No. No. X -A- R.sup.1 R.sup.2
R.sup.3 R.sup.4 R.sup.6 data 28 B.1 O --(CH.sub.2).sub.2-- 3-Cl H
4-Cl H OH mp. 248.degree. C. 29 B.1 O --(CH.sub.2).sub.3-- 3-Cl H
4-Cl H OH (3); mp. 154.degree. C. (3): ethanedioate (1:1) hydrate
(1:1)
[0136]
9TABLE F-3 21 Co. Ex. No. No. 22 -A- X R.sup.1 R.sup.2 R.sup.3
Physical data 36 B.11 double --(CH.sub.2).sub.3-- O 3-Cl H 4-Cl mp.
195.2.degree. C. 37 B.12 single --(CH.sub.2).sub.3-- O 3-Cl H 4-Cl
mp. 93.6.degree. C. 38 B.11 double --(CH.sub.2).sub.2-- O 3-Cl H
4-Cl mp. 204.1.degree. C. 39 B.13 double --(CH.sub.2).sub.2-- O
3-Cl H H mp. 200.degree. C. 40 B.13 double --(CH.sub.2).sub.2-- O
3-Cl H 2-Cl mp. 216.degree. C. 41 B.13 double --(CH.sub.2).sub.2--
O 3-Cl H 3-Cl mp. 180.degree. C. 42 B.13 double
--(CH.sub.2).sub.2-- O 3-Cl H 4-F mp. 210.degree. C. 43 B.13 double
--(CH.sub.2).sub.2-- O 3-Cl H 4-CH.sub.3 mp. 210.degree. C. 44 B.13
double --(CH.sub.2).sub.2-- O 3-Cl H 4-OCH.sub.3 mp. 120.degree. C.
45 B.13 double --(CH.sub.2).sub.2-- O 3-Cl 4-Cl 4-Cl mp.
210.degree. C. 46 B.13 double --(CH.sub.2).sub.2-- O 4-Cl H 4-Cl
mp. 190.degree. C.; (1) 47 B.13 double --(CH.sub.2).sub.2-- O
3-CH.sub.3 H 4-Cl mp. 200.degree. C. 48 B.13 double
--(CH.sub.2).sub.2-- O 3-Br H 4-Cl mp. 196.degree. C. 49 B.13
double --(CH.sub.2).sub.2-- O H H 4-Cl mp. 184.degree. C; (1) 50
B.14 double --(CH.sub.2).sub.2-- S 3-Cl H 4-Cl mp. 253.degree. C.
51 B.11 double --CH.dbd.CH-- O 3-Cl H 4-Cl mp. 104.degree. C. 52
B.13 double --(CH.sub.2*)--O-- O 3-Cl H 4-Cl mp. 208.degree. C. 53
B.13 double --(CH.sub.2*).sub.2--O-- O 3-Cl H 4-Cl mp. 180.degree.
C. *the CH.sub.2 moiety is linked to the nitrogen atom of the
2-quinolinone moiety (1): .ethanedioate (2:3) salt
[0137] C. Pharmacological Example
EXAMPLE C.1
[0138] "In Vitro Assay for Inhibition of Farnesyl Protein
Transferase"
[0139] Human farnesyl protein transferase was prepared essentially
as described (Y. Reiss et al., Methods: A Companion to Methods in
Enzymology, vol. 1, 241-245, 1990). Kirsten virus transformed human
osteosarcoma (KHOS) cells (American Type Culture Collection,
Rockville, Md., USA) grown as solid tumors in nude mice or grown as
monolayer cell cultures were used as a source of human enzyme.
Briefly, cells or tumors were homogenized in buffer containing 50
mM Tris, 1 mM EDTA, 1 mM EGTA and 0.2 mM
phenylmethylsulfonylfluoride (pH 7.5). The homogenates were
centrifuged 28,000.times.g for 60 min and the supernatants
collected. A 30-50% ammonium sulfate fraction was prepared, and the
resulting precipitate was resuspended in a small (10 to 20 ml)
volume of dialysis buffer containing 20 mM Tris, 1 mM
dithiothreitol and 20 .mu.M ZnCl.sub.2. The ammonium sulfate
fraction was dialyzed overnight against two changes of the same
buffer. The dialyzed material was applied to a 10.times.1 cm Q Fast
Flow Sepharose (Pharmacia LKB Biotechnology Inc., Piscataway, N.J.,
USA) which had been preequilibrated with 100 ml of dialysis buffer
supplemented with 0.05 M NaCl. The column was washed with an
additional 50 ml of dialysis buffer plus 0.05 M NaCl followed by a
gradient from 0.05 M to 0.25 M NaCl prepared in dialysis buffer.
The enzyme activity was eluted with a linear gradient of 0.25 to
1.0 M NaCl prepared in the dialysis buffer. Fractions containing 4
to 5 ml volumes of column eluate were collected and analyzed for
farnesyl protein transferase activity. Fractions with enzyme
activity were pooled and supplemented with 100 .mu.M ZnCl.sub.2.
Enzyme samples were stored frozen at -70.degree. C.
[0140] The activity of farnesyl protein transferase was measured
using the Farnesyl Transferase [.sup.3H] Scintillation Proximity
Assay (Amersham International plc., England) under the conditions
specified by the manufacturer. To assay for inhibitors of the
enzyme, 0.20 .mu.Ci of the [.sup.3H]-farnesylpyrophosphate
substrate and the biotinylated lamin B peptide substrate
(biotin-YRASNRSCAIM) were mixed with test compounds in a reaction
buffer consisting of 50 mM HEPES, 30 mM MgCl.sub.2, 20 mM KCl, 5 mM
dithiothreitol, 0.01% Triton X-100. Test compounds were delivered
in a 10 .mu.l volume of dimethylsulfoxide (DMSO) to achieve
concentrations of 1 and 10 .mu.g/ml in a final volume of 100 .mu.l.
The reaction mixture was warmed to 37.degree. C. The enzyme
reaction was started by adding 20 .mu.l of diluted human farnesyl
protein transferase. Sufficient enzyme preparation was added to
produce between 4000 to 15000 cpm of reaction product during the 60
min of reaction incubation at 37.degree. C. Reactions were
terminated by the addition of STOP/scintillation proximity bead
reagent (Amersham). The reaction product
[.sup.3H]-farnesyl-(Cys)-bi- otin lamin B peptide was captured on
the streptavidin linked scintillation proximity bead. The amount of
[.sup.3H]-farnesyl-(Cys)-biotin lamin B peptide synthesized in the
presence or absence of test compounds was quantified as cpm by
counting on a Wallac Model 1480 Microbeta Liquid Scintillation
Counter. The cpm of product was considered to be farnesyl protein
transferase activity. The protein farnesyl transferase activity
observed in the presence of test compound was normalized to
farnesyl transferase activity in the presence of 10% DMSO and
expressed as percent inhibition. In separate studies, some test
compounds exhibiting 50% or greater inhibition of farnesyl protein
transferase activity were evaluated for concentration-dependent
inhibition of enzyme activity. The effects of test compounds in
these studies were calculated as IC.sub.50 (concentration of test
compound producing 50% inhibition of enzyme activity) using the
LGIC50 computer program written by the Science Information Division
of R. W. Johnson Pharmaceutical Research Institute (Spring House,
Pa., USA) on a VAX computer. Compound 36 was found to have a
IC.sub.50 of 21 nM and compound 38 to have a IC.sub.50 of 15
nM.
EXAMPLE C.2
[0141] "Ras-Transformed Cell Phenotype Reversion Assay"
[0142] Insertion of activated oncogenes such as the mutant ras gene
into mouse NIH 3T3 cells converts the cells to a transformed
phenotype. The cells become tumorigenic, display anchorage
independent growth in semi-solid medium and lose contact
inhibition. Loss of contact inhibition produces cell cultures which
no longer form uniform monolayers. Rather, the cells pile up into
multicellular nodules and grow to very high saturation densities in
plastic tissue culture dishes. Agents such as protein farnesyl
transferase inhibitors which revert the ras transformed phenotype
restore the uniform monolayer growth pattern to cells in culture.
This reversion is easily monitored by counting the number of cells
in tissue culture plates. Transformed cells will achieve higher
cell numbers than cells which have reverted to an untransformed
phenotype. Compounds which revert the transformed phenotype should
produce antitumor effects in tumors bearing ras gene mutations.
[0143] Method:
[0144] Compounds are screened in tissue culture in NIH 3T3 cells
transformed by the T24 activated human H-ras gene. Cells are seeded
at an initial density of 200,000 cells per well (9.6 cm.sup.2
surface area) in six-well cluster tissue culture plates. Test
compounds are immediately added to 3.0 ml cell growth medium in a
3.0 .mu.l volume of DMSO, with a final concentration of DMSO in the
cell growth medium of 0.1%. The test compounds are run at
concentrations of 5, 10, 50, 100, and 500 nM along with a DMSO
treated vehicle control. (In case a high activity is observed at 5
nM, the test compound is tested at even lower concentrations.) The
cells are allowed to proliferate for 72 hours. Then the cells are
detached in 1.0 ml trypsin-EDTA cell dissociation medium and
counted on a Coulter particle counter.
[0145] Measurements:
[0146] Cell numbers expressed as cells per well are measured using
a Coulter Particle Counter.
[0147] All cell counts were corrected for the initial cell input
density by subtracting 200,000.
[0148] Control cell counts=[cell counts from cells incubated with
DMSO vehicle-200,000]
[0149] Test compound cell counts=[cell counts from cells incubated
with test compound-200,000]. 1 Test compound % inhibition = [ 1 -
test compound cell counts control cell counts ] .times. 100 % .
[0150] IC.sub.50 (i.e. the test compound concentration required to
inhibit enzyme activity by 50%) is calculated if sufficient data
are available, summarized in table C.2.
10 TABLE C.2 Co. No. IC.sub.50 (nM) 1 142 3 51 4 50 5 7.8 6 32 8
6.4 10 3.2 12 66 14 7.3 15 48 17 63 18 >500 19 240 21 18 22 352
23 442 24 12 25 37 26 21 27 37.1 28 100 29 184 30 373 31 >500 33
73.1 34 >500 35 >500 40 >500 41 >500 44 353 45 >500
48 >500 49 >500 51 >500 53 >500
EXAMPLE C.3
[0151] "Farnesyl Protein Transferase Inhibitor Secondary Tumor
Model"
[0152] The enzyme farnesyl protein transferase catalyzes the
covalent attachment of a farnesyl moiety derived from farnesyl
pyrophosphate to the oncogene product p21.sup.ras. This directs
p.sub.21.sup.ras to attach to plasma membranes. Once attached to
plasma membranes, mutant or oncogenic forms of p.sub.21.sup.ras
will provide a signal for the transformation and uncontrolled
growth of malignant tumor cells. Therefore, inhibitors of protein
farnesyltransferase will prevent the membrane attachment of
p21.sup.ras and inhibit growth of ras-transformed tumors.
[0153] Nude mice are inoculated with 1.times.10.sup.6 of T24
activated human H-ras gene transformed NIH 3T3 fibroblast cells
(T24 cells), subcutaneously in the inguinal region. After three
days to allow tumors to become established, treatment with test
compounds is begun via the oral route. The test compounds are
dissolved in a 20% .beta.-cyclodextrin in 0.1 N HCl solution and
administered orally as 0.1 ml of compound solution per 10 g mouse
body weight. Routinely used doses are 6.25, 12.5 and 25 mg/kg. Body
weights and tumor sizes are monitored during the ensuing 15 days of
treatment. At the end of treatment, animals are sacrificed and
tumors are weighed.
[0154] The "mean vehicle treated tumor weight" is defined as the
mean tumor weight from 10 to 15 mice treated with the test
compound.
[0155] The "mean tumor weight" is defined as the mean tumor weight
from 10 to 15 mice not treated with the test compound. 2 %
Reduction final tumor weight = [ 1 - mean tumor weight mean vehicle
treated tumor weight ] .times. 100 % .
11TABLE C.3 % reduction final Co. No. Dose tumor weight 8 6.25
mg/kg bid, po 41% 12.25 mg/kg bid, po 44% 25 mg/kg bid, po 49%
* * * * *