U.S. patent application number 12/760408 was filed with the patent office on 2010-12-30 for substituted quinolines for the treatment of cancer.
This patent application is currently assigned to CRYSTAX PHARMACEUTICALS, S.L.. Invention is credited to Juan Aymami Bofarull, Miquel Coll Capella, Amadeo Llebaria Soldevila, Isabel Navarro Munoz.
Application Number | 20100331355 12/760408 |
Document ID | / |
Family ID | 34639541 |
Filed Date | 2010-12-30 |
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United States Patent
Application |
20100331355 |
Kind Code |
A1 |
Aymami Bofarull; Juan ; et
al. |
December 30, 2010 |
Substituted Quinolines for the Treatment of Cancer
Abstract
Compounds of formula G.sub.1-L-G.sub.2, where G.sub.1, is a
radical structurally close to cryptolepine, -L- is a single
covalent bond or a covalent linking biradical selected from
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s-- and
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR''''(CH.sub.2).sub.t--,
--R''' and --R'''' are radicals, same or different, selected from
the group consisting of H and (C.sub.1-C.sub.3)-alkyl; r, s and t
are an integer from 1 to 3 and -G.sub.2 is H or a radical
structurally close to -G.sub.1, are intercalators. They are
compounds which intercalate between DNA base pairs, and are useful
as therapeutic agents against cancer, as assess by an in vitro test
of citotoxicity with human leukemia cells Jurkat E6-1 and human
carcinoma cells GLC-4. Preferred compounds are those where -G.sub.1
is bonded to -L- through a carbonyl amino and -L- is
--(CH.sub.2).sub.3NCH.sub.3(CH.sub.2).sub.3-- or
--(CH.sub.2).sub.2NCH.sub.3(CH.sub.2).sub.sNCH.sub.3(CH.sub.2).sub.2--
where s=2 or 3. -G.sub.1 is a radical selected from (IIa) and
(IIb); -G.sub.2 is a radical selected from H, a radical of formula
(IIa), a radical of formula (IIb), the N-radical of
1,8-naphthalimide, the C4-radical of 2-phenylquinoline and the
C9-radical of acridine. ##STR00001##
Inventors: |
Aymami Bofarull; Juan;
(Barcelona, ES) ; Coll Capella; Miquel; (Madrid,
ES) ; Llebaria Soldevila; Amadeo; (Madrid, ES)
; Navarro Munoz; Isabel; (Barcelona, ES) |
Correspondence
Address: |
BERENBAUM WEINSHIENK PC
370 17TH STREET, SUITE 4800
DENVER
CO
80202
US
|
Assignee: |
CRYSTAX PHARMACEUTICALS,
S.L.
Barcelona
ES
CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS
Madrid
ES
UNIVERSITAT POLITECNICA DE CATALUNYA
Barcelona
ES
|
Family ID: |
34639541 |
Appl. No.: |
12/760408 |
Filed: |
April 14, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10580140 |
May 19, 2006 |
7728000 |
|
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PCT/EP2004/013106 |
Nov 18, 2004 |
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12760408 |
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Current U.S.
Class: |
514/285 ;
546/70 |
Current CPC
Class: |
C07K 5/06026 20130101;
C07D 471/04 20130101; A61K 38/00 20130101; A61P 35/00 20180101 |
Class at
Publication: |
514/285 ;
546/70 |
International
Class: |
A61K 31/437 20060101
A61K031/437; C07D 471/04 20060101 C07D471/04; A61P 35/00 20060101
A61P035/00 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 20, 2003 |
ES |
P200302821 |
Claims
1. A compound of formula (I) G.sub.1-L-G.sub.2 (I) or a
pharmaceutically acceptable salt thereof, wherein: -G.sub.1 is a
radical (II) ##STR00014## wherein --R.sup.1 is an electron pair or
a (C.sub.1-C.sub.3)-alkyl radical; with the condition that (i) when
--R' is an electron pair, a is a N.dbd.C double bond and the fused
ring ##STR00015## is the biradical ##STR00016## thus radical (II)
is (IIa'), and ##STR00017## (ii) when --R' is a
(C.sub.1-C.sub.3)-alkyl radical, a is a N--C single bond and the
fused ring is the triradical ##STR00018## thus radical (II) is
(IIa''); ##STR00019## wherein --R.sub.1 to --R.sub.12 represent
radicals, same or different, selected from the group consisting of
H, (C.sub.1-C.sub.4)-alkyl, (C.sub.1-C.sub.4)-alkoxy,
(C.sub.1-C.sub.4)-alkylamino, phenyl, F, Cl, Br, amino, hydroxy,
and nitro; and wherein --B-- is a biradical selected from the group
consisting of --CONH--, --NR.sub.13--, --O--,
--(CH.sub.2).sub.nNH--, --(CH.sub.2).sub.nO--, and
--CO[NHCHR''CO].sub.mO--; wherein --R.sub.13 is selected from the
group consisting of H, (C.sub.1-C.sub.4)-alkyl,
(C.sub.1-C.sub.4)-alkoxy and (C.sub.1-C.sub.4)-alkylamino; --R''
are side chains radicals, same or different, corresponding to
natural aminoacids; n is an integer from 1 to 3 and m is an integer
from 1 to 3; -L- is a single covalent bond or a covalent linking
biradical selected from the following ones;
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s--
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR''''(CH.sub.2).sub.t--
wherein --R''' and --R'''' are radicals, same or different,
selected from the group consisting of H and
(C.sub.1-C.sub.3)-alkyl; r is an integer from 1 to 3; s is an
integer from 1 to 3; t is an integer from 1 to 3; and -G.sub.2 is a
radical selected from a radical of formula (II), the N-radical of
1,8-naphthalimide, the C4-radical of 2-phenylquinoline, and the
C9-radical of acridine.
2. The compound according to claim 1, wherein (II) is the radical
(IIa') ##STR00020##
3. The compound according to claim 2, wherein --B-- is selected
from the group consisting of --CONH-- and --NR.sub.13--.
4. The compound according to claim 2, wherein --B-- is
--CO[NHCHR''CO].sub.mO--.
5. The compound according to claim 4, wherein m=2, the leftward
--R'' is a glicine side chain, and the rightward --R'' is an
arginine side chain.
6. The compound according to claim 2, wherein -L- is a single
covalent bond.
7. The compound according to claim 2, wherein -L- is a covalent
linking biradical selected from the following ones
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s--
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR''''(CH.sub.2).sub.t--
8. The compound according to claim 7, wherein -L- is the biradical
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s--, --R''' is methyl, and
both r and s are 3.
9. The compound according to claim 7, wherein -L- is the covalent
linking biradical
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR''''(CH.sub.2).sub.t--- ,
both --R''' and --R'''' are methyl; both r and t are 2, and s is 2
or 3.
10. The compound according to claim 1, wherein (II) is the radical
(IIa'') ##STR00021##
11. The compound according to claim 10, wherein --B-- is selected
from the group consisiting of --CONH-- and --NR.sub.13--.
12. The compound according to claim 10, wherein --B-- is
--CO[NHCHR''CO].sub.mO--.
13. The compound according to claim 12, wherein m=2, the leftward
--R'' is a glicine side chain, and the rightward --R'' is the
arginine side chain.
14. The compound according to claim 10, wherein --R' is methyl.
15. The compound according to claim 14, wherein -L- is a single
covalent bond.
16. The compound according to claim 14, wherein -L- is a biradical
selected from the following ones.
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s--
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR''''(CH.sub.2).sub.t--
17. The compound according to claim 16, wherein -L- is the
biradical --(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s--, R''' is
methyl, and both r and s are 3.
18. The compound according to claim 16, wherein -L- is the
biradical
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR'''(CH.sub.2).sub.t--,
both --R''' and --R'''' are methyl; both r and t are 2, and s is an
integer from 2 to 3.
19. The compound according to claim 1, which is selected from the
group consisting of:
N-[3-[[3-[(9-acridinecarbonyl)amino]propyl]methylamino]propyl]-10H-indolo-
[3,2-b]quinoline-11-carboxamide (Ia);
N,N'-(4-methyl-4-azaheptamethylene)-di-(10H-indolo[3,2-b]quinoline-11,11'-
-carboxamide) (Ib);
N-[3-[3-[[2-(1,3-dioxo-(2,3-dihydro)-1H-benzo[de]isoquinolinyl]propyl]met-
hylamino]propyl]-10H-indolo[3,2-b]quinoline-11-carboxamide (Ic);
N-[3-[[3-[(2-phenyl-4-quinolinecarbonyl)amino]propyl]methylamino]propyl]--
10H-indolo[3,2-b]quinoline-11-carboxamide (Id);
N,N'-(3,7-dimethyl-3,7-diazanonamethylene)-di-(10H-indolo[3,2-b]quinoline-
-11,11'-carboxamide) (Ie);
N-[(9-acridinecarbonyl)-3,7,10-triaza-3,7-dimethyldecyl]-10H-indolo[3,2-b-
]quinoline-11-carboxamide (If);
N,N'-(3,6-dimethyl-3,6-diazaoctamethylene)-di-(10H-indolo[3,2-b]quinoline-
-11-11'-carboxamide (Ig);
N-[(9-acridinecarbonyl)-3,6-dimethyl-3,6-diazaoctamethylene]-10H-indolo[3-
,2-b]quinoline-11-carboxamide (Ih);
N-[[1,3-dioxo-(2,3-dihydro)-1H-benzo[de]isoquinolyl]-3,6-dimethyl-3,6-dia-
zaoctamethylene]-10H-indolo[3,2-b]quinoline-11-carboxamide (Ii);
N-[[1,3-dioxo-(2,3-dihydro)-1H-benzo[de]isoquinolyl]-3,7,10-triaza-3,7-di-
methyldecyl]-10H-indolo[3,2-b]quinoline-11-carboxamide (Ij);
N,N'-(4-methyl-4-azaheptamethylene)-di-(5-methyl-5H-indolo[3,2-b]quinolin-
e-11,11'-carboxamide) (Im);
N,N'-(4-methyl-4-azaheptamethylen)-di-(5-methyl-5H-indolo[3,2-b]quinoline-
-11,11'-amine (Iq);
N,N'-(3,7-dimethyl-3,7-diazanonamethylene)-di-(5-methyl-5H-indolo[3,2-b]q-
uinoline-11,11'-carboxamide) (Iy);
N,N'-(3,6-dimethyl-3,6-diazaoctamethylene)-di-(5-methyl-5H-indolo[3,2-b]q-
uinoline-11,11'-carboxamide) (Iz);
(3,7-diazanonamethylene)-di-(10H-indolo[3,2-b]quinoline-11,11'-carboxamid-
e (Iaa);
N,N'-(3,7-dimethyl-3,7-diazanonamethylene)-di-(5-methyl-5H-indolo-
[3,2-b]quinoline-11,11'-amine (Iab); and
N,N'-(3,6-dimethyl-3,6-diazaoctamethylene)-di-(5-methyl-5H-indolo[3,2-b]q-
uinoline-11,11'-amine (Iac).
20. A method for the preparation of a medicament for the treatment
of cancer which comprises administering to a subject a
therapeutically effective amount of a compound of formula (I)
G.sub.1-L-G.sub.2 (I) or a pharmaceutically acceptable salt
thereof, wherein: -G.sub.1 is a radical (II) ##STR00022## wherein
--R' is an electron pair or a (C.sub.1-C.sub.3)-alkyl radical; with
the condition that (i) when --R' is an electron pair, a is a
N.dbd.C double bond and the fused ring ##STR00023## is the
biradical ##STR00024## thus radical (II) is (IIa'), and
##STR00025## (ii) when --R' is a (C.sub.1-C.sub.3)-alkyl radical, a
is a N--C single bond and the fused ring is the triradical
##STR00026## thus radical (II) is (IIa''); ##STR00027## wherein
--R.sub.1 to --R.sub.12 represent radicals, same or different,
selected from the group consisting of H, (C.sub.1-C.sub.4)-alkyl,
(C.sub.1-C.sub.4)-alkoxy, (C.sub.1-C.sub.4)-alkylamino, phenyl, F,
Cl, Br, amino, hydroxy, and nitro; and wherein --B-- is a biradical
selected from the group consisting of --CONH--, --NR.sub.13--,
--O--,--(CH.sub.2).sub.nNH--, --(CH.sub.2).sub.nO--, and
--CO[NHCHR''CO].sub.mO--; wherein --R.sub.13 is selected from the
group consisting of H, (C.sub.1-C.sub.4)-alkyl,
(C.sub.1-C.sub.4)-alkoxy and (C.sub.1-C.sub.4)-alkylamino; --R''
are side chains radicals, same or different, corresponding to
natural aminoacids; n is an integer from 1 to 3 and m is an integer
from 1 to 3; -L- is a single covalent bond or a covalent linking
biradical selected from the following ones.
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s--
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR'''(CH.sub.2).sub.t--
wherein --R''' and --R''' are radicals, same or different, selected
from the group consisting of H and (C.sub.1-C.sub.3)-alkyl; r is an
integer from 1 to 3; s is an integer from 1 to 3; t is an integer
from 1 to 3; and -G.sub.2 is a radical selected from a radical of
formula (II), the N-radical of 1,8-naphthalimide, the C4-radical of
2-phenylquinoline, and the C9-radical of acridine.
21. A pharmaceutical composition comprising a therapeutically
effective amount of the compound as defined in claim 1, together
with appropriate amounts of pharmaceutical excipients or
carriers.
22. A method of producing a composition of matter of formula (I)
comprising one of the following processes: ##STR00028## when
biradical --B-- in -G.sub.1 is --CONH-- and -G.sub.2 is not an
N-radical of 1,8-naphtalimide; and wherein GP represents an amino
protective group and wherein formula (IV) is a monoprotected
bis-amine; or ##STR00029## when biradical --B-- in -G.sub.1 is
--CONH-- and -G.sub.2 is 1,8-naphtalimide; and wherein GP
represents an amino protective group and wherein formula (IV) is a
monoprotected bis-amine; or ##STR00030## when biradical --B-- is a
biradical selected from a group of: --NR.sub.13--, --O--,
--(CH.sub.2).sub.nNH, and --(CH.sub.2).sub.nO--; and wherein GP
represents an amino protective group.
Description
[0001] This invention relates to new compounds which intercalate
between DNA base pairs. Such compounds can be used for the
treatment of cancer.
BACKGROUND ART
[0002] Intercalators are a group of compounds that bind between the
DNA base pairs. This intercalation can produce an interruption of
transcription, replication and/or topoisomerase activity. It has
been found that many compounds that bind to DNA by intercalation
have antiproliferative properties and in vivo antitumour effects.
Intercalators have been used as anticancer drugs. Cryptolepine is a
naturally occurring alkaloid and it is used in traditional medicine
against malaria. Its formula, shown below, is structurally close to
some compounds of the present invention. Cryptolepine is able to
bind CG-rich sequences of DNA containing non-alternating CC sites,
and it has been reported as potential anticancer drug (cf. e.g. J.
N. Lisgarten et al., "The antimalarial and cytotoxic drug
cryptolepine intercalates into DNA at cytosine-cytosine sites",
Nature Structural Biology 2002, vol. 9, pp. 57-60). Thus, the
provision of new intercalators with anticancer activity is highly
desirable.
##STR00002##
SUMMARY OF THE INVENTION
[0003] According to an aspect of the present invention, there is
provided a compound of formula (I),
G.sub.1-L-G.sub.2 1. [0004] (ii) (I)
[0005] or a pharmaceutically acceptable salt thereof, wherein:
[0006] -G.sub.1 is a radical selected from (IIa) y (IIb);
##STR00003##
[0007] wherein --R' is an electron pair or a
(C.sub.1-C.sub.3)-alkyl radical; with the condition that [0008]
(iii) when --R' is an electron pair, a is a N.dbd.C double bond and
the
[0008] ##STR00004## fused ring
[0009] is the biradical
##STR00005##
[0010] thus radicals (IIa) and (IIb) are respectively (IIa') and
(II'), and
##STR00006## [0011] (iv) when --R' is a (C.sub.1-C.sub.3)-alkyl
radical, a is a N--C single bond and the fused ring
[0012] is the triradical
##STR00007##
[0013] thus radicals (IIa) and (IIb) are respectively (IIa'') and
(IIb'');
##STR00008##
[0014] wherein --R.sub.1 to --R.sub.12 represent radicals, same or
different, selected from the group consisting of H,
(C.sub.1-C.sub.4)-alkyl, (C.sub.1-C.sub.4)-alkoxy,
(C.sub.1-C.sub.4)-alkylamino, phenyl, F, Cl, Br, amino, hydroxy,
and nitro;
[0015] and wherein --B-- is a biradical selected from the group
consisting of --CONH--,
[0016] --NR.sub.13--, --O--, --(CH.sub.2).sub.nNH--,
--(CH.sub.2).sub.nO--, --CONH(CH.sub.2).sub.uZ--,
[0017] --CONH(CH.sub.2).sub.uCH(CH.sub.2OH)CH.sub.2Z-- and
--CO[NHCHR''CO].sub.mO--; wherein --R.sub.13 is selected from the
group consisting of H, (C.sub.1-C.sub.4)-alkyl,
(C.sub.1-C.sub.4)-alkoxy and
[0018] (C.sub.1-C.sub.4)-alkylamino; --R'' are side chains
radicals, same or different, corresponding to natural aminoacids; n
is an integer from 1 to 3; m is an integer from 1 to 3; u is an
integer from 1 to 3, and --Z-- is a biradical of a oligonucleotide
phosphate between 4 and 23 bases in length, linked to the methylene
group at the 5' end or at the 3' end;
[0019] -L- is a single covalent bond or a covalent linking
biradical selected from the following ones;
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s--
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR''''(CH.sub.2).sub.t--
[0020] wherein --R''' and --R'''' are radicals, same or different,
selected from the group consisting of H and
(C.sub.1-C.sub.3)-alkyl; r is an integer from 1 to 3; s is an
integer from 1 to 3; t is an integer from 1 to 3; and
[0021] -G.sub.2 is a radical selected from H, a radical of formula
(IIa), a radical of formula (IIb), the N-radical of
1,8-naphthalimide, the C4-radical of 2-phenylquinoline, and the
C9-radical of acridine;
[0022] with the proviso that (I) is not one of the compounds of the
following list, which have been previously described (the Chemical
Abstracts Registry number, CAS RN, is given) but not for the
industrial applicability of the present invention. The
corresponding values for radicals in formula (I) are also
included:
[0023] 10H-quindoline-11-carboxamide (CAS RN 367911-30-6;
-G.sub.1=IIa'; --B--.dbd.--CONH--; --R'=electron pair; --R.sub.1 to
R.sub.8.dbd.H; -L-=single covalent bond; -G.sub.2=--H);
[0024] 2-bromo-10H-quindoline-11-carboxamide (CAS RN 241470-56-4;
-G.sub.1=IIa'; --B--.dbd.--CONH--; --R'=electron pair;
--R.sub.2.dbd.--Br; --R.sub.1, --R.sub.3 to --R.sub.4, --R.sub.5 to
--R.sub.8.dbd.H; -L-=single covalent bond; -G.sub.2=--H);
[0025] N-10H-quindolin-11-yl-1,3-propanediamine (CAS RN
188630-48-0; -G.sub.1=IIa'; --B--.dbd.--NH--, --R'=electron pair;
--R.sub.1 to --R.sub.4, --R.sub.5 to --R.sub.8.dbd.--H;
-L-=--(CH.sub.2).sub.3NH--; -G.sub.2=--H);
[0026] 10H-quindolin-11-amine monohydrochloride (CAS RN
164406-49-9; G.sub.1=IIa', --B--.dbd.--NH; --R'=electron pair;
--R.sub.1 to --R.sub.4, --R.sub.5to --R.sub.8.dbd.--H; -L-=single
covalent bond; -G.sub.2=--H);
[0027] 10H-quindolin-11-methanol (CAS RN 241470-44-0; G.sub.1=IIa';
--B--.dbd.--CH.sub.2O--; --R'=electron pair; --R.sub.1 to
--R.sub.4, --R.sub.5 to --R.sub.8.dbd.H, -L-=single covalent bond,
-G.sub.2=--H); or
[0028] N-[2-(dimethylamino)ethyl]-10-H-quindoline-4-carboxamide
(CAS RN 191172-25-5; G.sub.1=IIb'; --B--.dbd.--CONH--;
--R'=electron pair; --R.sub.1 to --R.sub.4, --R.sub.5 to
--R.sub.8.dbd.--H, -L-=--(CH.sub.3).sub.2N(CH.sub.3)CH.sub.2--,
-G.sub.2=--H).
[0029] In a preferred embodiment, compounds of formula (I) are
those where (IIa) is the radical (IIa').
##STR00009##
[0030] Preferred compounds are those of formula (I) where (IIa) is
(IIa') and --B-- is
[0031] --CONH-- or --NR.sub.13--. Also preferred are those of
formula (I) where (IIa) is (IIa') and --B-- is
--CO[NHCHR''CO].sub.mO-- and more preferred are the compounds where
m=2, the leftward --R'' is a glicine side chain, and the rightward
--R'' is a arginine side chain . Also preferred compounds are those
of formula (I) where (IIa) is (IIa') and --B-- is
--CONH(CH.sub.2).sub.uZ-- or
--CONH(CH.sub.2).sub.uCH(CH.sub.2OH)CH.sub.2Z--. More preferred are
the compounds where --Z-- is -TTCCGGAA- linked at 3' end or at 5'
end, or -CTTCTTCTTCT- linked at 3' end. Specially preferred are
those where -L- is a single covalent bond. Also specially preferred
are those where -L- is a covalent linking biradical selected from
the following ones.
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s--
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR''''(CH.sub.2).sub.t--
[0032] The most preferred are the compounds where -L- is the
biradical --(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s--, --R''' is
methyl and both r and s are 3, and the compounds where -L- is the
biradical
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR''''(CH.sub.2).sub.t--,
--R''' and --R''' are methyl; both r and t are 2 and s is an
integer from 2 to 3.
[0033] In another preferred embodiment, compounds of formula (I)
are those where (IIa) is the following radical.
##STR00010##
[0034] Preferred compounds are those where (IIa) is (IIa'') and
--B-- is --CONH-- or
[0035] --NR.sub.13--. Also preferred are those where --B-- is
--CO[NHCHR''CO].sub.mO--, and more preferred are the compounds
where m=2, the leftward --R'' is a glicine side chain, and the
rightward --R'' is a arginine side chain. Also preferred compounds
are those of formula (I) where (IIa) is (IIa'') and --B-- is
--CONH(CH.sub.2).sub.uZ-- or
--CONH(CH.sub.2).sub.uCH(CH.sub.2OH)CH.sub.2Z--. More preferred are
the compounds where --Z-- is -TTCCGGAA- linked at 3' or 5' end or
-CTTCTTCTTCT- linked at 3' end. Even more preferred are those where
--R' is methyl. Specially preferred are those where -L- is a single
covalent bond. Also specially preferred are those where -L- is a
covalent linking biradical selected from the following ones.
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s--
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR''''CH.sub.2).sub.t--
[0036] The most preferred are the compounds where -L- is the
biradical --(CH.sub.2).sub.rNR'''(CH.sub.2).sub.s--, --R''' is
methyl and both r and s are 3, and the compounds where -L- is the
biradical
--(CH.sub.2).sub.rNR'''(CH.sub.2).sub.sNR''''(CH.sub.2).sub.t--,
--R''' and --R''' are methyl; both r and t are 2, and s is an
integer from 2 to 3.
[0037] The most preferred compounds of formula (I) are those of the
following list, whose preparation is described in the accompanying
examples for the first time:
N-[3-[[3-[(9-acridinecarbonyl)amino]propyl]methylamino]propyl]-10H-indolo-
[3,2-b]quinoline-11-carboxamide (Ia);
N,N'-(4-methyl-4-azaheptamethylene)-di-(10H-indolo[3,2-b]quinoline-11,11'-
-carboxamide) (Ib);
[0038]
N-[3-[3-[[2-(1,3-dioxo-(2,3-dihydro)-1H-benzo[de]isoquinolinyl)prop-
yl]methylamino]propyl]-10H-indolo[3,2-b]quinoline-11-carboxamide
(Ic);
N-[3-[[3-[2-phenyl-4-quinolinecarbonyl)amino]propyl]methylamino]propyl]-1-
0H-indolo[3,2-b]quinoline-11-carboxamide (Id);
N,N'-(3,7-dimethyl-3,7-diazanonamethylene)-di-(10H-indolo[3,2-b]quinoline-
-11,11'-carboxamide) (Ie);
[0039]
N-[(9-acridinecarbonyl)-3,7,10-triaza-3,7-dimethyldecyl]-10H-indolo-
[3,2-b]quinoline-11-carboxamide (If);
N,N'-(3,6-dimethyl-3,6-diazaoctamethylene)-di-(10H-indolo[3,2-b]quinoline-
-11-11'-carboxamide) (Ig);
N-[(9-acridinecarbonyl)-3,6-dimethyl-3,6-diazaoctamethylene]-10H-indolo[3-
,2-b]quinoline-11-carboxamide (Ih);
N-[[1,3-dioxo-(2,3-dihydro)-1H-benzo[b]isoquinolyl]-3,6-dimethyl-3,6-diaz-
aoctamethylene]-10H-indolo[3,2-b]quinoline-11-carboxamide (Ii);
N-[[1,3-dioxo-(2,3-dihydro)-1H-benzo[b]isoquinolyl]-3,7,10-triaza-3,7-dim-
ethyldecyl]-10H-indolo[3,2-b]quinoline-11-carboxamide (Ij);
N,N'-(4-methyl-4-azaheptamethylene)-di-(methyl-5H-indolo[3,2-b]quinoline--
11,11'-carboxamide) (Im);
10H-indolo[3,2-b]quinoline-11-carbonyl-glicine-arginine (Io);
N,N-dimethyl-N'-(5-methyl-5H-indolo[3,2-b]quinolin-11-yl)-ethane-1,2-diam-
ine (Ip);
N,N'-(4-methyl-4-azaheptamethylen)-di-(10H-indolo[3,2-b]quinolin-
e-11,11'-amine (Iq);
N-1-[(5-hydroxymethyl-6-(5'-TTCCGGAA-3'-phosphate)-hexyl]-10H-indolo[3,2--
b]quinoline-11-carboxamide (Ir);
N-1-[(5-hydroxymethyl-6-(5'-CTTCCTCCTCT-3'-phosphate)-hexyl]-10H-indolo[3-
,2-b]quinoline-11-carboxamide (Is); and
N-1-[6-(5'-phosphate-TTCCGGAA)-hexyl]-10H-indolo[3,2-b]quinoline-11-carbo-
xamide (It); 10H-Indolo[3,2-b]quinoline-11-carboxylic
acid(2-dimethylamino-ethyl)amide (Iu);
10H-Indolo[3,2-b]quinoline-11-carboxylic
acid(2-dimethylamino-propyl)amide (Iv);
N,N-dimethyl-N'-(5-methyl-5H-indolo[3,2-b]quinolin-11-yl)-propane-1,2-dia-
mine (Iw);
N-[3-[[3-[(10H-indolo[3,2-b]quinoline-4-carboxamide]propyl]meth-
ylamino]propyl]-10H-indolo[3,2-b]quinoline-11-carboxamide (Ix);
N,N'-(3,7-dimethyl-3,7-diazanonamethylene)-di-(methyl-5H-indolo[3,2-b]qui-
noline-11,11'-carboxamide) (Iy);
N,N'-(3,6-dimethyl-3,6-diazaoctamethylene)-di-(methyl-5H-indolo[3,2-b]qui-
noline-11,11'-carboxamide) (Iz);
(3,7-diazanonamethylene)-di-(10H-indolo[3,2-b]guinoline-11,11'-carboxamid-
e (Iaa);
N,N'-(3,7-dimethyl-3,7-diazanonamethylene)-di-(10H-indolo[3,2-b]g-
uinoline-11,11'-amine (lab);
N,N'-(3,6-dimethyl-3,6-diazaoctamethylene)-di-(10H-indolo[3,2-b]guinoline-
-11,11'-amine (lac).
[0040] Another aspect of the present invention refers to the use of
the compound of formula (I), defined above, for the preparation of
a medicament for the treatment of cancer.
[0041] A further aspect of the present invention relates to a
pharmaceutical composition comprising as active ingredient a
therapeutically effective amount of the compound of formula (I),
defined above, together with suitable pharmaceuticalexcipients or
carriers.
[0042] Compounds of formula (I) where biradical --B-- in -G.sub.1
is --CONH-- and -G.sub.2 is not the N-radical of 1,8-naphtalimide
can be prepared according to the process summarized in Scheme I,
where GP represents an amino protecting group. The process involves
a coupling reaction of a compound of formula (Vb) with a compound
of formula (III). The compound of formula (Vb) is prepared by
reaction of a compound of formula (III) with a monoprotected
bis-amine of formula (IV), followed by the removal of the amino
protective group. Monoprotected bis-amines of formula (IV) are
commercially available compounds or they can be prepared by methods
known in the art (cf. Spicer et al., Bioorg. Med. Chem. 2002, vol.
10, p. 19; Deady et al., Bioorg. Med. Chem. 2000, vol. 8, p. 977).
An example of amino protective group is the t-butoxycarbonyl
(BOC).
##STR00011##
[0043] When biradical --B-- in -G.sub.1 is --CONH-- and -G.sub.2 is
1,8-naphthalamide, compounds of formula (I) can be obtained by the
process summarized in Scheme II. This process involves the coupling
reaction of a compound of formula (VII.sub.b) with a compound of
formula (III). The compound of formula (VII.sub.b) can be prepared
by reaction of 1,8-naphthalic anhydride (VI) with a monoprotected
bis-amine of formula (IV), followed by the removal of the amino
protective group.
##STR00012##
[0044] Compounds of formula (I) where --B-- is a biradical selected
from other groups as --NR.sub.13--, --O--, --(CH.sub.2).sub.nNH--,
--(CH.sub.2).sub.nO--, can be obtained analogously by using
standards processes of organic chemistry, well known in the art for
the production of such type of compounds. For example, for
--B--.dbd.--NH-- the process of Scheme III can be followed, as it
has been done for the compound of the Example 15.
##STR00013##
[0045] Compounds of formula (I) where biradical --B-- is
--CO[NHCHR''CO].sub.mO--, --R'' is side chain radicals, same or
different, corresponding to a natural aminoacid and m is an integer
selected from 1 to 3, can be obtained by solid phase methods. The
synthesis is carried out assembling a N-protected aminoacid or
peptide in a p-methylbenzhydrilamine resine. After removal of the
protective group, the 11-aminocarboxylic radical selected from
formula (IIa) or the 4-aminocarboxylic radical selected from
formula (IIb) is coupled by
benzotriazol-1-yloxytripyrrolidinophosphonium hexafluoro-phosphate
(PyBOP) and diisopropylamine, followed by acidolysis.
[0046] Compounds of formula (I) where --B-- is a birradical
selected from --CONH(CH.sub.2).sub.uZ-- or
--CONH(CH.sub.2).sub.uCH(CH.sub.2OH)CH.sub.2Z--, where u is an
integer from 1 to 6, and --Z-- is a biradical of a oligonucleotide
phosphate between 4 and 23 bases in length, linked to the methylene
group at 5' end or at 3' end can be prepared by conventional
methods of peptide coupling in solid phase or solution phase.
[0047] Compounds of formula (I) may be converted into
pharmaceutically acceptable acid addition salts, and salts may be
converted into free compounds, by conventional methods. For
instance, the acid addition salts may be prepared by contacting the
free base with an appropriate amount of the desired acid in a
conventional manner.
[0048] Compounds of formula (I) are intercalators (i.e. compounds
which intercalate between DNA base pairs) and, as illustrated by
biological results in example 30 are active for cancer
treatment.
[0049] Throughout the description and claims the word "comprise"
and variations of the word, are not intended to exclude other
technical features, additives, components, or steps. The abstract
of this application is incorporated herein as reference. Additional
objects, advantages and features of the invention will become
apparent to those skilled in the art upon examination of the
description or may be learned by practice of the invention. The
following examples are provided by way of illustration, and they
are not intended to be limiting of the present invention
EXAMPLES
Example 1
Preparation of
N-[3-[[3-[(9-acridinecarbonyl)amino]propyl]methylamino]propyl]-10H-indolo-
[3,2-b]quinoline-11-carboxamide (Ia)
[0050] 10H-indolo[3,2-b]quinoline-11-carboxylic acid previously
prepared (cf. Bierer et al., J. Med. Chem. 1998, vol. 41, p. 894
(0.2 g, 0.76 mmol) was dissolved in 5 ml of refluxing SOCl.sub.2.
After 6 hours a clear solution was obtained, and all volatiles were
removed in vacuo. The crude acid chloride was suspended in 5 ml of
CH.sub.2Cl.sub.2. A solution of
[3-[(3-aminopropyl)methylamino]propyl]carbamic acid tert-butyl
ester previously prepared (Spicer at el., Bioorg. Med. Chem. 2002,
vol. 10, p. 19) (0.26 g, 1.1 mmol) and triethylamine (0.4 ml) in 5
ml of CH.sub.2Cl.sub.2 was added at room temperature, and the
mixture was stirred for 14 hours. The organic phase was washed
(NaHCO.sub.3, brine) and dried (MgSO.sub.4). Removal of the solvent
and purification by chromatography on alumina
(CH.sub.2Cl.sub.2/MeOH, 90:10) yielded
[2-[[2-[(10H-Indolo[3,2-b]quinoline-11-carbonyl)amino]ethyl]methylamino)e-
thyl]carbamic acid tert-butyl ester (0.26 g, 70%) as a viscous oil.
.sup.1H-NMR [MeOD, .delta., ppm]: 8.48 (d, J=7.8 Hz, 1H), 7.25 (m,
2H), 7.64 (m, 4H), 7.35 (dd, J=8.0 Hz, 1H), 3.67 (t, J=6.9 Hz, 2H,
CH.sub.2NO), 3.07 (t, J=6.9 Hz, 2H, CH.sub.2NHBOC), 2.56 (m, 2H,
CH.sub.2N(CH.sub.3)), 2.43 (m, 2H, CH.sub.2N(CH.sub.3)), 2.26 (s,
3H, N(CH.sub.3)), 1.66 (m, 4H, 2CH.sub.2CN(CH.sub.3)), 1.48 (s, 9H,
C(CH.sub.3).sub.3).
[0051] To a solution of
[2-[[2-[(10H-indolo[3,2-b]quinoline-11-carbonyl)amino]-ethyl]methylamino)-
ethyl]-carbamic acid tert-butyl ester (0.13 g, 0.27 mmol) in
CH.sub.2Cl.sub.2 (15 ml), was added trifluoroacetic acid (3 ml).
The reaction mixture was stirred at room temperature for 16 h at
which point the reaction was completed by TLC. All solvents were
removed under reduced pressure to give
10H-indolo[3,2-b]quinoline-[3-[(3-aminopropyl)methylamino]propyl]-11-carb-
oxamide (0.09 g, 92%) as an oil, which was used directly in the
next reaction. .sup.1H-NMR [MeOD, .delta., ppm]: 8.51 (d, J=7.8 Hz,
1H), 8.32 (m, 2H), 8.02 (m, 1H), 7.84 (m, 2H), 7.67 (d, J=8.0 Hz,
1H), 7.44 (m, 1H), 3.76 (t, J=6.9 Hz, 2H, CH.sub.2NO), 3.31 (m, 2H,
CH.sub.2NH.sub.2), 2.96 (s, 3H, N(CH.sub.3)), 2.27 (m, 2H,
CH.sub.2N(CH.sub.3)), 2.15 (m, 2H, CH.sub.2N(CH.sub.3)), 1.26 (m,
4H, 2CH.sub.2CN(CH.sub.3)).
[0052] 9-acridinecarboxylic acid (0.036 g, 0.16 mmol) was dissolved
in 3 ml of refluxing SOCl.sub.2. After 6 hours a clear solution was
obtained, and all volatiles were removed in vacuo. The crude acid
chloride was suspended in 4 ml of CH.sub.2Cl.sub.2. A solution of
10H-indolo[3,2-b]quinoline-[3-[(3-aminopropyl)methyl-amino]propyl]-11-car-
boxamide (0.076 g, 0.19 mmol) and triethylamine (0.2 ml) in 4 ml of
CH.sub.2Cl.sub.2 was added at room temperature, and the reaction
mixture was stirred for 14 hours. The organic phase was washed
(NaHCO.sub.3, brine) and dried (MgSO.sub.4). Removal of the solvent
and purification by chromatography on alumina
(CH.sub.2Cl.sub.2/MeOH, 95:5) yielded the desired compound (55 mg,
50%) as a solid. mp 108-110.degree. C. .sup.1H-NMR [CDCl.sub.3,
.delta., ppm]: 8.38 (m, 1H), 8.05 (m, 7H), 7.58 (m, 7H), 7.26 (m,
1H), 3.59 (m, 4H, 2CH.sub.2NO), 2.55 (m, 4H, 2CH.sub.2NO), 2.26 (s,
3H, N(CH.sub.3)), 1.91 (m, 4H, 2CH.sub.2CN(CH.sub.3)); MS (EI, m/z)
595 (M.sup.++1).
Example 2
Preparation of
N,N'-(4-methyl-4-azaheptamethylene)-di-(10H-indolo[3,2-b]quinoline-11,11'-
-carboxamide (Ib)
[0053] 10H-indolo[3,2-b]quinoline-11-carboxylic acid (0.02 g, 0.08
mmol) was dissolved in 2 ml of refluxing SOCl.sub.2. After 6 hours
a clear solution was obtained, and all volatiles were removed in
vacuo. The crude acid chloride was suspended in 2 ml of
CH.sub.2Cl.sub.2. A solution of
10H-indolo[3,2-b]quinoline-[3-[(3-aminopropyl)methylamino]propyl]-11-carb-
oxamide (0.037 g, 0.09 mmol) prepared as in Example 1, and
triethylamine (0.2 ml) in 2 ml of CH.sub.2Cl.sub.2 was added at
room temperature, and the mixture was stirred for 14 hours. The
organic phase was washed (NaHCO.sub.3, brine) and dried
(MgSO.sub.4). Removal of the solvent and purification by
chromatography on alumina (CH.sub.2Cl.sub.2/MeOH, 95:5) yielded the
desired compound (30 mg, 50%) as a solid. mp 124-126.degree. C.
.sup.1H NMR [CDCl.sub.3, .delta., ppm]: 8.47 (m, 2H), 8.17 (m, 4H),
7.59 (m, 8H), 7.25 (m, 2H), 3.65 (m, 4H, 2CH.sub.2NO), 2.63 (m, 4H,
2CH.sub.2NO), 2.33 (s, 3H, N(CH.sub.3)), 1.99 (m, 4H,
2CH.sub.2CN(CH.sub.3)); MS (EI, m/z) 634 (M.sup.++1).
Example 3
Preparation of
N-[3-[3-[2-[1,3-dioxo-(2,3-dihydro)-1H-benzo[de]isoquinolinyl]propyl]meth-
ylamino]propyl]-10H-indolo[3,2-b]quinoline-11-carboxamide (Ic)
[0054] 1,8-naphthalic anhydride (0.18 g, 0.9 mmol) was added to a
solution of [3-[(3-aminopropyl)methylamino]propyl]carbamic acid
tert-butyl ester previously prepared (Spicer at el., Bioorg. Med.
Chem. 2002, vol. 10, p. 19) (0.2 g, 0.8 mmol) in absolute ethanol
(20 ml). The reaction mixture was heated at reflux overnight.
Removal of the solvent and purification by chromatography on
alumina (CH.sub.2Cl.sub.2/MeOH, 50:1) yielded
[3-[[3-(1,3-dioxo-3a,9b-dihydro-1H,3H-benzo[de]isoquinolin-2-yl)propyl]me-
thylamino]propyl]carbamic acid tert-butyl ester (0.37 g, 90%) as a
foam. .sup.1H-NMR [CDCl.sub.3, .delta., ppm]: 8.62 (m, 2H), 8.22
(m, 2H), 7.72 (m, 2H), 4.23 (m, 2H, 2CH.sub.2NO), 3.20 (m, 2H,
CH.sub.2NH), 2.55 (m, 4H, 2(CH.sub.2)), 2.26 (s, 3H, N(CH.sub.3)),
1.93 (m, 2H, (CH.sub.2)), 1.68 (m, 2H, (CH.sub.2)), 1.48 (s, 9H,
C(CH.sub.3).sub.3).
[0055] To a solution of
[3-[[3-(1,3-dioxo-3a,9b-dihydro-1H,3H-benzo[de]isoquinolin-2-yl)propyl]me-
thylamino]propyl]carbamic acid tert-butyl ester (0.34 g, 0.81 mmol)
in CH.sub.2Cl.sub.2 (24 ml) was added trifluoroacetic acid (6 ml).
The reaction mixture was stirred at room temperature for 16 h at
which point the reaction was completed by TLC. All solvents were
removed under reduced pressure to give
2-[3-[(3-aminopropyl)methylamino]propyl]-3a,9b-dihydro-benzo[de]isoquinol-
ine-1,3-dione (0.24 g, 92%) as an oil, which was used directly in
the next reaction. .sup.1H-NMR [MeOD, .delta., ppm]: 8.64 (m, 2H),
8.54 (m, 2H), 7.92 (m, 2H), 4.45 (m, 2H, 2CH.sub.2NO), 3.55 (m, 4H,
2CH.sub.2), 3.33 (m, 2H, CH.sub.2), 3.22 (s, 3H, N(CH.sub.3)), 2.44
(m, 4H, 2(CH.sub.2)).
[0056] 10H-indolo[3,2-b]quinoline-11-carboxylic acid (0.24 g, 0.9
mmol) was dissolved in 7 ml of refluxing SOCl.sub.2. After 6 hours
a clear solution was obtained, and all volatiles were removed in
vacuo. The crude acid chloride was suspended in 10 ml of
CH.sub.2Cl.sub.2. A solution of
2-[3-[(3-aminopropyl)methyl-amino]propyl]-3a,9b-dihydro-benzo[de]isoquino-
line-1,3-dione (0.24 g, 0.73 mmol) and NEt.sub.3 (1 ml) in 10 ml of
CH.sub.2Cl.sub.2 was added at room temperature, and the reaction
mixture was stirred for 14 hours. The organic phase was washed
(NaHCO.sub.3, brine) and dried (MgSO.sub.4). Removal of the solvent
and chromatography on alumina (CH.sub.2Cl.sub.2/MeOH, 95:5) yielded
the desired compound (0.13 g, 30%) as a solid. mp 200-204.degree.
C. .sup.1H-NMR [CDCl.sub.3, .delta., ppm]: 8.38-8.02 (m, 7H),
7.61-7.37 (m, 7H), 3.86 (m, 4H, 2CH.sub.2NO), 2.60 and 2.40 (m, 4H,
2CH.sub.2NO), 2.22 (s, 3H, N(CH.sub.3)), 1.91 and 1.61 (m, 4H,
2CH.sub.2CN(CH.sub.3)).
Example 4
Preparation of
N-[3-[[3-[2-phenyl-4-quinolinecarbonyl)amino]propyl]methylamino]propyl]-1-
0H-indolo[3,2-b]quinoline-11-carboxamide (Id)
[0057] Activation and coupling of
10H-indolo[3,2-b]quinoline-11-carboxylic acid with
2-phenyl-quinoline-[3-[(3-aminopropyl)methylamino]propyl]-4-car-
boxamide (prepared analogously to Example 1 from
2-phenyl-4-quinolinecarboxylic acid), gave the desired compound as
a solid, 36%, mp 144-148.degree. C. .sup.1H-NMR [MeOD, .delta.,
ppm]: 8.18 (m, 2H), 8.08 (m, 4H), 7.61-7.44 (m, 10H), 7.28 (m, 1H),
3.68 (m, 2H, 2CH.sub.2NO), 3.54 (m, 2H, 2CH.sub.2NO), 3.34 (s, 3H,
N(CH.sub.3)), 2.64 (m, 4H, 2CH.sub.2CN(CH.sub.3)); 1.98 (m, 4H,
2CH.sub.2); MS (EI, m/z) 619 (M.sup.+-1).
Example 5
Preparation of
N,N'-(3,7-dimethyl-3,7-diazanonamethylene)-di-(10H-indolo[3,2-b]quinoline-
-11,11'-carboxamide) (Ie)
[0058] 10H-indolo[3,2-b]quinoline-11-carboxylic acid (0.25 g, 0.95
mmol) was dissolved in 5 ml of refluxing SOCl.sub.2. After 6 hours
a clear solution was obtained, and all volatiles were removed in
vacuo. The crude acid chloride was suspended in 5 ml of
CH.sub.2Cl.sub.2. A solution of
[2-[[3-[(2-aminoethyl)methylamino]propyl]methylamino]ethyl]carbamic
acid tert-butyl ester (Deady et al., Bioorg. Med. Chem. 2000, vol.
8, p. 977) (0.38 g, 1.3 mmol) and triethylamine (0.4 ml) in 5 ml of
CH.sub.2Cl.sub.2 was added at room temperature. The reaction
mixture was stirred for 14 hours. The organic phase was washed
(NaHCO.sub.3, brine) and dried (MgSO.sub.4). Removal of the solvent
and purification by chromatography on alumina
(CH.sub.2Cl.sub.2/MeOH, 90:10) yielded
[2-[[3-[[2-[(10H-indolo[3,2-b]quinoline-11-carbonyl)-amino]ethyl]methylam-
ino]propyl]methyl-amino]ethyl]carbamic acid tert-butyl ester (0.31
g, 62%) as a viscous oil . .sup.1H-NMR [CDCl.sub.3, .delta., ppm]:
8.45 (d, J=7.8 Hz, 1H), 8.25 (m, 2H), 7.62 (m, 4H), 7.35 (dd,
J=8.0, 8.0 Hz, 1H), 3.85 (m, 2H, CH.sub.2NO), 3.21 (m, 2H,
CH.sub.2), 3.07 (m, 2H, CH.sub.2NHBOC), 2.79 (m, 4H,
2CH.sub.2N(CH.sub.3)), 2.37-2.19 (m, 4H, 2(CH.sub.2)), 2.15 (s, 6H,
2N(CH.sub.3)), 1.48 (s, 9H, C(CH.sub.3).sub.3).
[0059] To a solution
[2-[[3-[[2-[(10H-indolo[3,2-b]quinoline-11-carbonyl)amino]ethyl]methylami-
no]propyl]methylamino]ethyl]carbamic acid tert-butyl ester (0.3 g,
0.57 mmol) in CH.sub.2Cl.sub.2 (20 ml), was added trifluoroacetic
acid (6 ml). The reaction mixture was stirred at room temperature
for 16 h, at which point the reaction was completed by TLC. All
solvents were removed under reduced pressure to give
10H-indolo[3,2-b]quinoline-[2-[[3-[(2-aminoethyl)methylamino]propyl]methy-
lamino]ethyl]-11-carboxamide (0.23 g, 92%) as a oil, which was used
directly. .sup.1H-NMR [MeOD, .delta., ppm]: 8.59 (d, J=7.8 Hz, 1H),
8.44 (m, 2H), 8.09 (m,1H), 7.94 (m, 2H), 7.87 (d, J=8.0 Hz, 1H),
7.51 (m, 1H), 3.68 (m, 2H, CH.sub.2NO), 3.46 (m, 8H, 4CH.sub.2),
3.11 (m, 4H, 2CH.sub.2N(CH.sub.3)), 2.97 (s, 6H, 2N(CH.sub.3)).
[0060] 10H-indolo[3,2-b]quinoline-11-carboxylic acid (0.11 g, 0.41
mmol) was dissolved in 10 ml of refluxing SOCl.sub.2. After 6 hours
a clear solution was obtained, and all volatiles were removed in
vacuo. The crude acid chloride was suspended in 15 ml of
CH.sub.2Cl.sub.2. A solution of
10H-indolo[3,2-b]quinoline-11-carboxylic
acid[2-[[3-[(2-aminoethyl)methylamino]propyl]methylamino]-ethyl]-amide
(0.23 g, 0.5 mmol) and triethylamine (0.5 ml) in 15 ml of
CH.sub.2Cl.sub.2 was added at room temperature. The reaction
mixture was stirred for 14 hours. The organic phase was washed
(NaHCO.sub.3, brine) and dried (MgSO.sub.4). Removal of the solvent
and purification by chromatography on alumina
(CH.sub.2Cl.sub.2/MeOH, 95:5) yielded the desired compound (0.14 g,
52%) as a foam. .sup.1H-NMR [CDCl.sub.3, .delta., ppm]: 8.48 (m,
2H), 8.30 (m, 4H), 7.73-7.54 (m, 8H), 7.25 (m, 2H) 3.77 (m, 4H,
2CH.sub.2NO), 2.58 (m, 4H, 2CH.sub.2N(CH.sub.3)), 2.38 (m, 6H,
4CH.sub.2), 2.15 (s, 6H, 2N(CH.sub.3)).
Example 6
Preparation of
N-[(9-acridinecarbonyl)-3,7,10-triaza-3,7-dimethyldecyl]-10H-indolo[3,2-b-
]quinoline-11-carboxamide (If)
[0061] 9-acridinecarboxylic acid (0.15 g, 0.67 mmol) was dissolved
in 15 ml of refluxing SOCl.sub.2. After 6 hours a clear solution
was obtained, and all volatiles were removed in vacuo. The crude
acid chloride was suspended in 15 ml of CH.sub.2Cl.sub.2. A
solution of
10H-indolo[3,2-b]quinoline-[2-[[3-[(2-aminoethyl)methylamino]propyl]methy-
lamino]ethyl]-11-carboxamide (0.4 g, 0.92 mmol) and triethylamine
(1 ml) in 15 ml of CH.sub.2Cl.sub.2 was added at room temperature.
The reaction mixture was stirred for 14 hours. The organic phase
was washed (NaHCO.sub.3, brine) and dried (MgSO.sub.4). Removal of
the solvent and purification by chromatography on alumina
(CH.sub.2Cl.sub.2/MeOH, 95:5) yielded the desired compound (0.38 g,
88%) as a foam. .sup.1H-NMR [CDCl.sub.3, .delta., ppm]: 8.12 (m,
1H), 8.02 (m, 7H), 7.58 (m, 7H), 7.33 (m, 1H), 3.77 (m, 4H,
2CH.sub.2NO), 2.58 (m, 4H, 2CH.sub.2N(CH.sub.3)), 2.48 (m, 6H,
4CH.sub.2), 2.28 (s, 6H, 2N(CH.sub.3)).
Example 7
Preparation of
N,N'-(3,6-dimethyl-3,6-diazaoctamethylene)-di-(10H-indolo[3,2-b]quinoline-
-11-11'-carboxamide (Ig)
[0062] Activation and coupling of
10H-indolo[3,2-b]quinoline-11-carboxylic acid with
10H-indolo[3,2-b]quinoline-[2-[[2-[(2-aminoethyl)methylamino]-e-
thyl]-methylamino]thyl]-11-carboxamide (prepared analogously to
Example 5 from
[2-[[2-[(2-aminoethyl)methylamino]ethyl]methylamino]ethyl]carbamic
acid tert-butyl ester) gave the desired compound as a foam, 38%.
.sup.1H-NMR [CDCl.sub.3, .delta., ppm]: 8.46 (m, 2H), 8.30 (m, 4H),
7.73-7.54 (m, 8H), 7.18 (m, 2H) 3.62 (m, 4H, 2CH.sub.2NO), 2.58 (m,
4H, 2CH.sub.2N(CH.sub.3)), 2.38 (m, 4H, 4CH.sub.2), 2.35 (s, 6H,
2N(CH.sub.3)).
Example 8
Preparation of
N-[(9-acridinecarbonyl)-3,6-dimethyl-3,6-diazaoctamethylene)-10H-indolo[3-
,2-b]quinoline-11-carboxamide (Ih)
[0063] Activation and coupling of 9-acridinecarboxylic acid with
10H-indolo[3,2-b]quinoline-[2-[[2-[(2-aminoethyl)methylamino]ethyl]methyl-
amino]ethyl]-11-carboxamide (prepared analogously to Example 5 from
[2-[[2-[(2-aminoethyl)methylamino]ethyl]methylamino]ethyl]carbamic
acid tert-butyl ester) gave the desired compound as a foam, 48%.
.sup.1H-NMR [CDCl.sub.3, .delta., ppm]: 8.12 (m, 1H), 8.02 (m, 7H),
7.58 (m, 7H), 7.33 (m, 1H), 4.02 (m, 4H, 2CH.sub.2NO), 3.98 (m, 4H,
2CH.sub.2N(CH.sub.3)), 3.01 (m, 4H, 4CH.sub.2), 2.70 (s, 6H,
2N(CH.sub.3)).
Example 9
Preparation of
N-[[1,3-dioxo-(2,3-dihydro)-1H-benzo[de]isoquinolyl]-3,6-dimethyl-3,6-dia-
zaoctamethylene]-10H-indolo[3,2-b]quinoline-11-carboxamide (Ii)
[0064] Activation and coupling of
10H-indolo[3,2-b]quinoline-11-carboxylic acid with
2-[2-[[2-[(2-aminoethyl)methylamino]ethyl]methylamino]ethyl]-be-
nzo[de]isoquinoline-1,3-dione (prepared analogously to Example 3
from
2-[[2-[(2-amino-ethyl)methylamino]ethyl]methylamino]ethyl]carbamic
acid tert-butyl ester) gave the desired compound as a foam, 37%,
.sup.1H-NMR [CDCl.sub.3, .delta., ppm]: 8.55-8.11 (m, 7H),
7.72-7.17 (m, 7H), 4.03 (m, 4H, 2CH.sub.2NO), 3.80 (m, 4H,
2CH.sub.2N(CH.sub.3)), 2.93-2.64 (m, 4H, 4CH.sub.2), 2.55 (s, 6H,
2N(CH.sub.3)).
Example 10
Preparation of
N-[[1,3-dioxo-(2,3-dihydro)-1H-benzo[de]isoquinolyl]-3,7,10-triaza-3,7-di-
methyldecyl]-10H-indolo[3,2-b]quinoline-11-carboxamide (Ij)
[0065] Activation and coupling of
10H-indolo[3,2-b]quinoline-11-carboxylic acid with
2-[2-[[3-[(2-aminoethyl)methylamino]propyl]methylamino]ethyl]-b-
enzo[de]isoquinoline-1,3-dione (prepared analogously to Example 3
from
2-[[3-[(2-aminoethyl)methylamino]propyl]methylamino]ethyl]carbamic
acid tert-butyl ester) gave the desired compound as a foam, 40%.
.sup.1H-NMR [CDCl.sub.3 .delta., ppm]: 8.59-8.02 (m, 7H), 7.77-7.25
(m, 7H), 4.05 (m, 4H, 2CH.sub.2NO), 3.82 (m, 4H,
2CH.sub.2N(CH.sub.3)), 2.78-2.34 (m, 6H, 4CH.sub.2), 2.25 (s, 6H,
2N(CH.sub.3)).
Example 11
Preparation of 5-methyl-5H-indolo[3,2-b]quinoline-11-carboxylic
acid, Intermediate for the Preparation of (Im)
[0066] The 5-methyl-5H-indolo[3,2-b]quinoline-11-carboxylic acid is
known in the literature as part of extract plant Cryptolepis
sanguinolenta (cf. Paulo et al., Planta Medica 2000, vol. 66, p.
30). It can also be prepared by the following process:
[0067] 5,11-dimethyl-5H-indolo[3,2-b]quinoline (cf. Yang et al., J.
Nat. Prod, 1999, vol. 62, pp. 976] (0.1 g, 0.4 mmol) was added to
10 ml of water. KMnO.sub.4 (64 mg. 0.4 mmol) is added, and the
solution is heated to reflux for 1 h. Then a second portion of
KMnO.sub.4 (64 mg. 0.4 mmol) is introduced, followed by 10 ml of
water, and the heating is continued for 1 h. The reaction mixture
is allowed to cool slightly, and the precipitated oxide of
manganese is filtered and washed with hot water. The filtrated is
concentrated under reduced pressure to half of the volume, filtered
and acidified with concentrated hydrochloric acid. This acid
solution is then evaporated to dryness under reduced pressure to
give the desired compound (70 mg, 64%), which was used directly.
.sup.1H-NMR [MeOD .delta., ppm]: 8.24 (m, 2H), 7.79 (m, 2H), 7.54
(m, 4H), 5.15 (s, 3H) MS (EI, m/z) 277 (M.sup.++1).
Example 12
Preparation of
N,N'-(4-methyl-4-azaheptamethylene)-di-(5-methyl-5H-indolo[3,2-b]quinolin-
e-11,11'-carboxamide) (Im)
[0068] Activation and coupling of
5-methyl-5H-indolo[3,2-b]quinoline-11-carboxylic acid, prepared as
in Example 11, with
5-methyl-5H-indolo[3,2-b]quinoline-[3-[(3-aminopropyl)methylamino]propyl]-
-11carboxamide (prepared analogously to Example 1 from
5-methyl-5H-indolo[3,2-b]quinoline-11-carboxylic acid) gave the
desired compound as a foam, 50%, .sup.1H NMR [CDCl.sub.3 .delta.,
ppm]: 8.47 (m, 2H), 8.17 (m, 4H), 7.59 (m, 8H), 7.25 (m, 2H), 5.15
(s, 6H), 3.65 (m, 4H, 2CH.sub.2NO), 2.63 (m, 4H, 2CH.sub.2NO), 2.33
(s, 3H, N(CH.sub.3)), 1.99 (m, 4H, 2CH.sub.2CN(CH.sub.3)).
Example 13
Preparation of
10H-indolo[3,2-b]quinoline-11-carbonyl-glicine-arginine (Io)
[0069] The compound of the title was prepared by solid phase
methods. After assembling the Boc-glicine-arginine sequence on a
p-methylbenzhydrylamine resin, the N-terminal was deprotected and
10H-indolo[3,2-b]quinoline-11-carboxylic acid was coupled by means
of benzotriazol-1-yloxytripyrrolidinophosphonium
hexafluoro-phosphate (PyBOP) and diisopropylethylamine (5, 5 and 10
equivalents, respectively) in 1:1 DMSO-DMF for 1 h. Acidolysis with
HF/anisole (9:1, 0.degree. C., 1 h) provided the desired compound
in highly homogeneous form (>98% by HPLC). MS(MALDI-TOF):
EM(expected): 474.1; EM(found): 475.5 (M+H.sup.+).
Example 14
Preparation of
N,N-dimethyl-N'-(5-methyl-5H-indolo[3,2-b]quinolin-11-yl)-ethane-1,2-diam-
ine (Ip)
[0070] A solution of 11-cloro-5-metil-5H-indolo[3,2-b]quinoline
previously prepared (cf. Bierer et al., J. Med. Chem. 1998, vol.
41, p. 2754) (66 mg, 0.25 mmol) and N,N-dimethylethylenediamine (94
.mu.l, 1 mmol) in 2-ethoxyethanol (10 ml) was heated at 120.degree.
C. for 10 min. Removal of the solvent and chromatography on alumina
(CHCl.sub.3 100%) yielded the desired compound (36 mg, 46%) as a
yellow solid. m. p. 162-164.degree. C. .sup.1H NMR [CDCl.sub.3
.delta., ppm]: 8.28 (d, J=8.5 Hz, 1H), 7.89 (m, 2H), 7.69-7.61 (m,
3H), 7.31 (m, 2H), 4.58 (s, 3H, NCH.sub.3), 4.32 (m, 2H, CH.sub.2),
3.14 (m, 2H, CH.sub.2), 2.51 (s, 6H, N(CH.sub.3).sub.2).
Example 15
Preparation of
N,N'-(4-methyl-4-azaheptamethylen)-di-(5-methyl-5H-indolo[3,2-b]quinoline-
-11,11'-amine (Iq)
[0071] A solution of 11-chloro-5-methyl-5H-indolo[3,2-b]quinoline
(cf. Bierer et al., J. Med. Chem. 1998, vol. 41, pp. 2754) (61 mg,
0.23 mmol) and [[3-[(3-amino-propyl)methylamino]propyl]carbamic
acid tert-butyl ester previously prepared (Spicer at el., Bioorg.
Med. Chem. 2002, vol. 10, p. 19) (84 .mu.l, 0.34 mmol) in
2-ethoxyethanol (10 ml) was heated at 120.degree. C. for 10 min.
Removal of the solvent and purification by chromatography on
alumina (CHCl.sub.3: MeOH 2%) yielded
[3-[methyl-[3-(5-methyl-5H-indolo[3,2-b]quinolin-11-yl-amino)propyl]-amin-
o]propyl]carbamic acid tert-butyl ester (49 mg, 49%) as a foam.
.sup.1H NMR [CDCl.sub.3 .delta., ppm]: 8.16 (d, J=8.5 Hz, 1H), 7.81
(m, 4H), 7.55-7.48 (m, 2H), 7.21 (m, 1H), 4.51 (s, 3H, NCH.sub.3),
4.27 (m, 2H, CH.sub.2NO), 3.12 (m, 2H, CH.sub.2NHBOC), 2.76 (m, 2H,
CH.sub.2N(CH.sub.3)), 2.63 (m, 2H, CH.sub.2N(CH.sub.3)), 2.39 (s,
3H, N(CH.sub.3)), 2.34 (m, 2H, CH.sub.2CN(CH.sub.3)), 1.73 (m, 2H,
CH.sub.2CN(CH.sub.3)), 1.37 (s, 9H, C(CH.sub.3).sub.3).
[0072] To a solution of
[3-[methyl-[3-(5-methyl-5H-indolo[3,2-b]quinolin-11-ylamino)-propyl]amino-
]propyl]-carbamic acid tert-butyl ester (49 mg, 0.1 mmol) in
CH.sub.2Cl.sub.2 (10 ml), was added trifluoroacetic acid (12
.mu.l). This mixture was stirred at room temperature for 16 h, at
which point the reaction was completed by TLC. All solvents were
removed under reduced pressure to give
N.sup.1-methyl-N.sup.1-[3-(5-methyl-5H-indolo[3,2-b]quinolin-11-ylamino)p-
ropyl]propane-1,3-diamine (38 mg, 92%) as an oil, which was used
directly. .sup.1H-NMR [MeOD, .delta., ppm]: 8.52 (d, J=8.5 Hz, 1H),
8.35-7.73 (m, 4H), 7.63-7.60 (m, 2H), 7.31 (m, 1H), 4.45 (s, 3H,
NCH.sub.3), 3.60 (m, 2H, CH.sub.2NO), 3.02 (m, 2H,
CH.sub.2N(CH.sub.3)), 2.92 (s, 3H, N(CH.sub.3)), 2.36 (m, 4H,
2CH.sub.2CN(CH.sub.3)), 2.14 (m, 2H, CH.sub.2N(CH.sub.3)), 1.90 (m,
2H, CH.sub.2CN(CH.sub.3)).
[0073] A solution of 11-chloro-5-methyl-5H-indolo[3,2-b]quinoline,
previously prepared, (cf. Bierer et al., J. Med. Chem. 1998, vol.
41, p. 2754) (40 mg, 0.15 mmol) and
N.sup.1-methyl-N.sup.1-[3-(5-methyl-5H-indolo[3,2-b]quinolin-11-ylamino)--
propyl]propane-1,3-diamine (38 mg, 0.1 mmol) in 2-ethoxyethanol (10
ml) was heated at 120.degree. C. for 10 min. Removal of the solvent
and purification by chromatography on alumina (CHCl.sub.3 100%)
yielded the desired compound (17 mg, 46%) as a foam. .sup.1H NMR
[CDCl.sub.3 .delta., ppm]: 8.16-8.21 (m, 4H), 8.01 (m, 2H),
7.93-7.71 (m, 4H), 7.50-7.30 (m, 2H), 7.23-7.25 (m, 4H), 4.26 (s,
3H, NCH.sub.3), 4.24 (s, 3H, NCH.sub.3), 4.12 (m, 2H, CH.sub.2NO),
3.03 (m, 2H, CH.sub.2), 2.80 (m, 2H, CH.sub.2N(CH.sub.3)), 2.70 (m,
2H, CH.sub.2N(CH.sub.3)), 2.44 (s, 3H, N(CH.sub.3)), 2.16 (m, 2H,
CH.sub.2CN(CH.sub.3)), 1.72 (m, 2H, CH.sub.2CN(CH.sub.3)).
Example 16
Preparation of
N-1-[5-hydroxymethyl-6-(5'-TTCCGGAA-3'-phosphate)-hexyl]-10H-indolo[3,2-b-
]quinoline-11-carboxamide by Solid Phase Coupling (Ir)
[0074] 5'TTCCGGAA-NH.sub.2-3' oligonucleotide sequence carrying an
amino group at the 3' end was prepared using
3-O-dimethoxytrityloxy-2-(N-9-fluorenphenylmethoxycarbonyl-4-aminobutyl)--
1-propanyl ester succinamidyl-controlled pore glass (3' amino C7
modifier CPG from Glen Research). The solid support obtained after
the assembly of the sequence was treated with a 0.1 M solution of
1,8-diazabiciclo[5.4.0]undece-7-ene (DBU) in dry acetonitrile (2
min, room temperature). In this way, the Fmoc group that protects
the amino group is removed selectively with a non-nucleophillic
base. The resulting support was washed with acetonitrile and
reacted with 10H-indolo[3,2-b]quinoline-11-carboxylic acid as
follows. A mixture containing 10 molar excess of
10H-indolo[3,2-b]quinoline-11-carboxylic acid, 20 molar excess of
diisopropylethylamine (DIEA) as base catalyst and 10 molar excess
of (benzotriazol-1-yloxy)trispyrrolidinophosphonium
hexafluorophosphate (PyBOP) as carboxyl activator was prepared in
dry dimethylformamide (DMF) (0.2 ml). The mixture was left for 2
min at room temperature and added to the support. After 1 hour at
room temperature, the mixture was filtered and washed with DMF. The
support was dried and concentrated ammonia (1 ml) was added. The
ammonia solution was left for 1 hr at 50.degree. C. The mixture was
filtered and the ammonia solution was concentrated to dryness. The
residue was dissolved in water and desalted by NAP-10 (Sephadex
G-25) eluted with water. Finally, the oligonucleotide fractions
were analyzed by HPLC. HPLC solutions were as follows. Solvent A:
5% ACN in 100 mM triethylammonium acetate pH 6.5 and solvent B: 70%
ACN in 100 mM triethylammonium acetate pH 6.5. Columns: PRP-1
(Hamilton), 250.times.10 mm. Flow rate: 3 ml/min. A 20 min linear
gradient from 5-35%. The compound of the title eluted at 14.0 min.
Unreacted amino-oligonucleotide eluted around 9 min. The desired
product was characterized by the UV-spectra. UVmax: 258,348. Yield:
7%.
Example 17
Preparation of
N-1-[5-hydroxymethyl-6-(5'-TTCCGGAA-3'-phosphate)-hexyl]-10H-indolo[3,2-b-
]quinoline-11-carboxamide by Solution-Phase Coupling (Ir)
[0075] 5'TTCCGGAA-NH.sub.2 3' oligonucleotide sequence carrying an
amino group at the 3' end was prepared using the 3' amino C7
modifier CPG (Glen Research). The solid support obtained after the
assembly of the sequence was treated with concentrated ammonia for
1 hr at 50.degree. C. The mixture was filtered and the ammonia
solution was concentrated to dryness. The residue was passed over a
Dowex 50.times.4 (Na+ form) column to exchange ammonium ions for
sodium ions. The resulting amino-oligonucleotide was dissolved in
0.1 ml of water, and mixed with 0.1 ml of 1M sodium carbonate
aqueous buffer pH 8-9. In a separate container
10H-indolo[3,2-b]quinoline-11-carboxylic acid (10 molar excess) was
dissolved in 0.1 ml of DMF, and mixed with N-hydroxysuccinimide (10
molar excess) and N,N-diisopropylcarbodiimide (10 molar excess).
The mixture was left for 10 min at room temperature and added to
the aqueous solution of the amino-oligonucleotide. The reaction
mixture was kept at room temperature overnight. The mixture was
concentrated to dryness and the residue was dissolved in water. The
solution was passed through a NAP-10 column. The fractions
containing oligonucleotide were analyzed by HPLC. HPLC solutions
were as follows: Solvent A: 5% ACN in 100 mM triethylammonium
acetate pH 6.5 and solvent B: 70% ACN in 100 mM triethylammonium
acetate pH 6.5. Columns: PRP-1 (Hamilton), 250.times.10 mm. Flow
rate: 3 ml/min. A 20 min linear gradient from 5-35%. The compound
of the title eluted at 14.0 min Unreacted amino-oligonucleotide
eluted around 9 min. The desired product was characterized by the
UV-spectra. UVmax: 258, 348 and mass spectrometry (MALDI-TOF):
found 2861 (M+H.sup.+) expected 2862. Yield: 18%.
Example 18
Preparation of
N-1-[5-hydroxymethyl-6-(5'-CTTCCTCCTCT-3'-phosphate)-hexyl]-10H-indolo[3,-
2-b]quinoline-11-carboxamide by Solid Phase Coupling (Is)
[0076] The compound of the title was obtained in a similar way to
example 16 but ammonia treatment was run for 2 hours at room
temperature. The compound of the title eluted at 13.8 min. It was
characterized by the UV-spectra. UVmax: 273, 349. Yield: 5%.
Example 19
Preparation of
N-1-[5-hydroxymethyl-6-(5'-CTTCCTCCTCT-3'-phosphate)-hexyl]-10H-indolo[3,-
2-b]quinoline-11-carboxamide by Solution-Phase Coupling (Is)
[0077] The compound of the title was obtained in a similar way to
example 17. The compound of the title eluted at 13.9 min. It was
characterized by the UV-spectra. UVmax: 273, 349, and mass
spectrometry (MALDI-TOF): found 3646 (M+H.sup.+) expected 3647.
Yield: 20%.
Example 20
Preparation of
N-1-[6-(5'-phosphate-TTCCGGAA)-hexyl]-10H-indolo[3,2-b]quinoline-11-carbo-
xamide by Solution-Phase Coupling (It)
[0078] 5'NH.sub.2-TTCCGGAA 3' oligonucleotide sequence carrying an
amino group at the 5' end was prepared using the phosphoramidite of
the N6-monomethoxytrityl (MMT) protected derivative of
6-aminohexanol (Glen Research). The solid support obtained after
the assembly of the sequence was treated with concentrated ammonia
for 1 hr at 50.degree. C. Reaction of the 5' amino-oligonucleotide
with 10H-indolo[3,2-b]quinoline-11-carboxylic acid was carried out
as described in example 17. The compound of the title eluted at 16
min. The desired product was characterized by the UV-spectra UVmax:
268, 349. Yield: 12%.
Example 21
Preparation of 10H-Indolo[3,2-b]quinoline-11-carboxylic
acid(2-dimethylamino-ethyl)amide (Iu)
[0079] 10H-Indolo[3,2-b]quinoline-11-carboxylic acid (0.5 g, 1.9
mmol) was dissolved in 20 ml of refluxing SOCl.sub.2. After 6 hours
a clear solution was obtained, and all volatiles were removed in
vacuo. The crude acid chloride was suspended in 10 ml of
CH.sub.2Cl.sub.2. A solution of N,N-dimethylethylendiamine (0.2 mL,
2.26 mmol) and triethylamine (2.2 ml) in 10 ml of CH.sub.2Cl.sub.2
was added at room temperature, and the mixture was stirred for 14
hours. The organic phase was washed (NaHCO.sub.3, brine) and dried
(MgSO.sub.4). Removal of the solvent and purification by
chromatography on alumina (CH.sub.2Cl.sub.2/MeOH, 90:10) yielded
the desired compound (0.38 g, 61%) as a solid. m. p.
175-176.degree. C. .sup.1H-NMR [MeOD, .delta., ppm]: 8.16 (d, J=7.8
Hz, 1H), 7.92 (m, 2H), 7.35-7.20 (m, 3H), 7.04 (m, 2H), 3.62 (m,
2H, CH.sub.2NO), 2.55 (m, 2H, CH.sub.2N(CH.sub.3).sub.2), 2.29 (s,
6H, N(CH.sub.3).sub.2). MS (EI, m/z) 277 (M.sup.++1).
Example 22
Preparation of 10H-Indolo[3,2-b]quinoline-11-carboxylic
acid(2-dimethylamino-propyl)amide (Iv)
[0080] 10H-Indolo[3,2-b]quinoline-11-carboxylic acid (0.2 g, 0.76
mmol) was dissolved in 8 ml of refluxing SOCl.sub.2. After 6 hours
a clear solution was obtained, and all volatiles were removed in
vacuo. The crude acid chloride was suspended in 10 ml of
CH.sub.2Cl.sub.2. A solution of N,N-dimethylproylendiamine (0.1 mL,
0.97 mmol) and triethylamine (1 ml) in 10 ml of CH.sub.2Cl.sub.2
was added at room temperature, and the mixture was stirred for 14
hours. The organic phase was washed (NaHCO.sub.3, brine) and dried
(MgSO.sub.4). Removal of the solvent and purification by
chromatography on alumina (CH.sub.2Cl.sub.2/MeOH, 90:10) yielded
the desired compound (80 mg, 31%) as a solid. m. p. 188-189.degree.
C. .sup.1H-NMR [MeOD, .delta., ppm]: 8.16 (d, J=7.8 Hz, 1H), 7.92
(m, 2H), 7.35-7.20 (m, 3H), 7.04 (m, 2H), 3.62 (m, 2H, CH.sub.2NO),
2.55 (m, 2H, CH.sub.2N(CH.sub.3).sub.2), 2.29 (s, 6H,
N(CH.sub.3).sub.2), 1.88 (m, 2H, CH.sub.2). MS (EI, m/z) 347
(M.sup.++1).
Example 23
Preparation of
N,N-dimethyl-N'-(5-methyl-5H-indolo[3,2-b]quinolin-11-yl)-propane-1,2-dia-
mine (Iw)
[0081] A solution of 11-chloro-5-metil-5H-indolo[3,2-b]quinoline
(47 mg, 0.17 mmol) and N,N-dimethylpropylenediamine (78 .mu.l, 0.76
mmol) in 2-ethoxyethanol (5 ml) was heated at 120.degree. C. for 10
min. Removal of the solvent and chromatography on alumina
(CHCl.sub.3 100%) yielded the desired compound (20 mg, 35%) as a
foam. .sup.1H NMR [CDCl.sub.3 .delta., ppm]: 8.28 (d, J=8.5 Hz,
1H), 7.89 (m, 2H), 7.69-7.61 (m, 3H), 7.31 (m, 2H), 4.58 (s, 3H,
NCH.sub.3), 4.23 (m, 2H, CH.sub.2), 3.86 (m, 2H, CH.sub.2), 3.55
(m, 2H, CH.sub.2), 2.55 (s, 6H, N(CH.sub.3).sub.2). MS (EI, m/z)
333 (M.sup.++1).
Example 24
Preparation of
N-[3-[[3-[(10H-indolo[3,2-b]quinoline-4-carboxamide]propyl]methylamino]pr-
opyl]-10H-indolo[3,2-b]quinoline-11-carboxamide (Ix)
[0082] 10H-indolo[3,2-b]quinoline-4-carboxylic acid (0.2 g, 0.76
mmol) was dissolved in 5 ml of refluxing SOCl.sub.2. After 6 hours
a clear solution was obtained, and all volatiles were removed in
vacuo. The crude acid chloride was suspended in 5 ml of
CH.sub.2Cl.sub.2. A solution of
[3-[(3-aminopropyl)methylamino]propyl]carbamic acid tert-butyl
ester (0.26 g, 1.1 mmol) and triethylamine (0.4 ml) in 5 ml of
CH.sub.2Cl.sub.2 was added at room temperature, and the mixture was
stirred for 14 hours. The organic phase was washed (NaHCO.sub.3,
brine, HCl 1N) and dried (MgSO.sub.4). All solvents were removed
under reduced pressure to give
10H-indolo[3,2-b]quinoline-[3-[(3-aminopropyl)methylamino]propyl]-4-carbo-
xamide (0.14 g, 48%) as an oil, which was used directly in the next
reaction. .sup.1H-NMR [MeOD, .delta., ppm]: 8.72 (d, J=7.8 Hz, 1H),
8.63 (d, J=7.8 Hz, 1H), 8.51 (d, J=7.8 Hz, 1H), 7.97 (m, 2H), 7.79
(d, J=8.0 Hz, 1H), 7.43 (m, 2H), 3.75 (t, J=6.9 Hz, 2H,
CH.sub.2NO), 3.31 (m, 2H, CH.sub.2NH.sub.2), 2.96 (s, 3H,
N(CH.sub.3)), 2.27 (m, 2H, CH.sub.2N(CH.sub.3)), 2.15 (m, 2H,
CH.sub.2N(CH.sub.3)), 1.35 (m, 4H, 2CH.sub.2CN(CH.sub.3)).
[0083] 10H-indolo[3,2-b]quinoline-11-carboxylic acid (0.066 g, 0.25
mmol) was dissolved in 3 ml of refluxing SOCl.sub.2. After 6 hours
a clear solution was obtained, and all volatiles were removed in
vacuo. The crude acid chloride was suspended in 2 ml of
CH.sub.2Cl.sub.2. A solution of
10H-indolo[3,2-b]quinoline-[3-[(3-aminopropyl)methyl-amino]propyl]-4-carb-
oxamide (0.14 g, 0.36 mmol) and triethylamine (0.2 ml) in 2 ml of
CH.sub.2Cl.sub.2 was added at room temperature, and the reaction
mixture was stirred for 14 hours. The organic phase was washed
(NaHCO.sub.3, brine) and dried (MgSO.sub.4). Removal of the solvent
and purification by chromatography on alumina
(CH.sub.2Cl.sub.2/MeOH, 95:5) yielded the desired compound (27 mg,
17%) as an oil. .sup.1H-NMR [CDCl3, 6, ppm]: 8.43 (m, 1H), 8.28 (m,
7H), 7.58 (m, 7H), 7.26 (m, 1H), 3.20 (m, 4H, 2CH.sub.2NO), 2.49
(m, 4H, 2CH.sub.2NO), 2.05 (s, 3H, N(CH.sub.3)), 1.64 (m, 4H,
2CH.sub.2CN(CH.sub.3)).
Example 25
Preparation of
N,N'-(3,7-dimethyl-3,7-diazanonamethylene)-di-(5-methyl-5H-indolo[3,2-b]q-
uinoline-11,11'-carboxamide) (Iy)
[0084] Activation and coupling of
5-methyl-5H-indolo[3,2-b]quinoline-11-carboxylic acid with
5-methyl-5H-indolo[3,2-b]quinoline[2-[[2-[(2-aminoethyl)methylamino]-ethy-
l]-methylamino]ethyl]-11-carboxamide (prepared analogously to
Example 5 from
[2-[[3-[(2-aminoethyl)methylamino]propyl]methylamino]ethyl]carbamic
acid tert-butyl ester and
5-methyl-5H-indolo[3,2-b]quinoline-11-carboxylic acid gave the
desired compound as a foam, (4 mg, 15%), .sup.1H NMR [CDCl.sub.3
.delta., ppm]: 8.47 (m, 2H), 8.17 (m, 4H), 7.59 (m, 8H), 7.25 (m,
2H), 5.15 (s, 6H), 3.77 (m, 4H, 2CH.sub.2NO), 2.58 (m, 4H,
2CH.sub.2N(CH.sub.3)), 2.38 (m, 6H, 4CH.sub.2), 2.15 (s, 6H,
2N(CH.sub.3)).
Example 26
Preparation of
N,N'-(3,6-dimethyl-3,6-diazaoctamethylene)-di-(5-methyl-5H-indolo[3,2-b]q-
uinoline-11,11'-carboxamide) (Iz)
[0085] Activation and coupling of
5-methyl-5H-indolo[3,2-b]quinoline-11-carboxylic acid with
5-methyl-5H-indolo[3,2-b]quinoline[2-[[2-[(2-aminoethyl)methylamino]-ethy-
l]-methyl-amino]ethyl]-11-carboxamide (prepared analogously to
Example 7 from
[2-[[2-[(2-aminoethyl)methylamino]ethyl]methylamino]ethyl]carbamic
acid tert-butyl ester and
5-methyl-5H-indolo[3,2-b]quinoline-11-carboxylic acid gave the
desired compound as a foam, (28 mg, 10%), .sup.1H NMR [CDCl.sub.3
.delta., ppm]: 8.47 (m, 2H), 8.17 (m, 4H), 7.59 (m, 8H), 7.25 (m,
2H), 5.15 (s, 6H), 3.65 (m, 4H, 2CH.sub.2NO), 2.58 (m, 4H,
2CH.sub.2N(CH.sub.3)), 2.38 (m, 4H, 4CH.sub.2), 2.35 (s, 6H,
2N(CH.sub.3)).
Example 27
Preparation of
(3,7-diazanonamethylene)-di-(10H-indolo[3,2-b]quinoline-11,11'-carboxamid-
e (Iaa)
[0086] 10H-indolo[3,2-b]quinoline-11-carboxylic acid (0.056 g, 0.21
mmol) was dissolved in 2 ml of refluxing SOCl.sub.2. After 6 hours
a clear solution was obtained, and all volatiles were removed in
vacuo. The crude acid chloride was suspended in 2 ml of
CH.sub.2Cl.sub.2. A solution of
(2-[3-(2-amino-ethylamino]-ethyl)-carbamic acid tert-butyl ester
(0.079 g, 0.3 mmol) and triethylamine (0.2 ml) in 2 ml of
CH.sub.2Cl.sub.2 was added at room temperature. The reaction
mixture was stirred for 14 hours. The organic phase was washed
(NaHCO.sub.3, brine) and dried (MgSO.sub.4). Removal of the solvent
and purification by chromatography on alumina
(CH.sub.2Cl.sub.2/MeOH, 90:10) yielded
[2-(3-{2-[10H-indolo[3,2-b]quinoline-11-carbonyl)-amino]-ethylamino}-prop-
ylamino)-ethyl]carbamic acid tert-butyl ester (0.025 g, 25%) as a
viscous oil . .sup.1H-NMR [CDCl.sub.3, .delta., ppm]: 8.46 (d,
J=7.8 Hz, 1H), 8.30 (m, 2H), 7.68 (m, 4H), 7.53 (dd, J=8.0, 8.0 Hz,
1H), 3.65 (m, 2H, CH.sub.2NO), 3.12 (m, 2H, CH.sub.2), 3.07 (m, 2H,
CH.sub.2NHBOC), 2.06 (m, 4H, 2CH.sub.2NH), 1.67-1.55 (m, 4H,
2(CH.sub.2)), 1.48 (s, 9H, C(CH.sub.3).sub.3).
[0087] To a solution
[2-(3-{2-[10H-indolo[3,2-b]quinoline-11-carbonyl)-amino]-ethylamino}-prop-
ylamino)-ethyl]carbamic acid tert-butyl ester (0.025 g, 0.049 mmol)
in CH.sub.2Cl.sub.2 (5 ml), was added trifluoroacetic acid (1 ml).
The reaction mixture was stirred at room temperature for 16 h, at
which point the reaction was completed by TLC. All solvents were
removed under reduced pressure to give
10H-indolo[3,2-b]quinoline-11-carboxylic
acid(2-[3-(2-amino-ethylamino)-propylamino]-ethyl]-amide (0.02 g ,
95%) as a oil, which was used directly. .sup.1H-NMR [MeOD, .delta.,
ppm]: 8.55 (d, J=7.8 Hz, 1H), 8.26 (m, 2H), 8.00 (m,1H), 7.80 (m,
2H), 7.64 (d, J=8.0 Hz, 1H), 7.26 (m, 1H), 4.19 (m, 2H,
CH.sub.2NO), 3.56 (m, 8H, 4CH.sub.2), 3.20 (m, 4H,
2CH.sub.2NH).
[0088] 10H-indolo[3,2-b]quinoline-11-carboxylic acid (0.009 g,
0.034 mmol) was dissolved in 2 ml of refluxing SOCl.sub.2. After 6
hours a clear solution was obtained, and all volatiles were removed
in vacuo. The crude acid chloride was suspended in 2 ml of
CH.sub.2Cl.sub.2. A solution of
10H-indolo[3,2-b]quinoline-11-carboxylic
acid(2-[3-(2-amino-ethylamino)-propylamino]-ethyl]-amide (0.02 g,
0.049 mmol) and triethylamine (0.2 ml) in 2 ml of CH.sub.2Cl.sub.2
was added at room temperature. The reaction mixture was stirred for
14 hours. The organic phase was washed (NaHCO.sub.3, brine) and
dried (MgSO.sub.4). Removal of the solvent and purification by
chromatography on alumina (CH.sub.2Cl.sub.2/MeOH, 95:5) yielded the
desired compound (0.015 g, 68%) as a foam. .sup.1H-NMR [CDCl.sub.3,
.delta., ppm]: 8.56 (m, 2H), 8.30 (m, 4H), 7.88-7.65 (m, 8H), 7.15
(m, 2H) 3.62 (m, 4H, 2CH.sub.2NO), 2.37 (m, 4H, 2CH.sub.2NH), 2.17
(m, 6H, 4CH.sub.2).
Example 28
Preparation of
N,N'-(3,7-dimethyl-3,7-diazanonamethylene)-di-(5-methyl-5H-indolo[3,2-b]q-
uinoline-11,11'-amine (Iab)
[0089] Activation and coupling of
11-chloro-5-methyl-5H-indolo[3,2-b]quinoline with
N-(2-amino-ethyl)-(N,N'-dimethyl-N'-[2-(5-methyl-5H-indolo[3,2-b]quinolin-
-11-ylamino)-ethyl]-propane-1,3-diamine (prepared analogously to
Example 15 from
[2-[[3-[(2-aminoethyl)methylamino]propyl]methylamino]ethyl]carbam-
ic acid tert-butyl ester and
11-chloro-5-methyl-5H-indolo[3,2-b]quinoline gave the desired
compound as a foam, (13 mg, 16%), .sup.1H NMR [CDCl.sub.3 .delta.,
ppm]: 8.58-8.40 (m, 4H), 8.01 (m, 2H), 7.84-7.64 (m, 4H), 7.63-7.55
(m, 2H), 7.42-7.27 (m, 4H), 4.61 (s, 6H, NCH.sub.3), 4.12 (m, 4H,
2CH.sub.2NH), 2.58 (m, 4H, 2CH.sub.2N(CH.sub.3)), 2.38 (m, 6H,
4CH.sub.2), 2.15 (s, 6H, 2N(CH.sub.3)).
Example 29
Preparation of
N,N'-(3,6-dimethyl-3,6-diazaoctamethylene)-di-(5-methyl-5H-indolo[3,2-b]q-
uinoline-11,11'-amine (Iac)
[0090] Activation and coupling of
11-chloro-5-methyl-5H-indolo[3,2-b]quinoline with
N-(2-amino-ethyl)-(N,N'-dimethyl-N'-[2-(5-methyl-5H-indolo[3,2-b]quinolin-
-11-ylamino)-ethyl]-ethane-1,2-diamine (prepared analogously to
Example 15 from
[2-[[2-[(2-aminoethyl)methylamino]ethyl]methylamino]ethyl]carbamic
acid tert-butyl ester and
11-chloro-5-methyl-5H-indolo[3,2-b]quinoline gave the desired
compound as a foam, (23 mg, 56%), .sup.1H NMR [CDCl.sub.3 .delta.,
ppm]: 8.66-8.21 (m, 4H), 8.01 (m, 2H), 7.84-7.64 (m, 4H), 7.63-7.55
(m, 2H), 7.42-7.27 (m, 4H), 4.91 (s, 6H, NCH.sub.3), 4.12 (m, 4H,
2CH.sub.2NH), 2.58 (m, 4H, 2CH.sub.2N(CH.sub.3)), 2.38 (m, 4H,
4CH.sub.2), 2.35 (s, 6H, 2N(CH.sub.3)).
Example 30
In Vitro Biological Test
[0091] The in vitro cytotoxicity of the compounds of the present
invention was evaluated by colorimetric assays with tetrazol salts
(MTT) on Jurkat clon E6-1 and on GLC-4, human leukemia and
carcinoma cell lines, respectively. This assay is based on the
capacity of live cells to incorporate and reduce the MTT, with
yellow colour in a red derivative. This reduction is done by the
action of the mitochondrial enzyme succinate hydrogenase, only
active in live cells. The intensity of observed colour is directly
correlated with the quantity of live cells in the sample as it is
described by Mosmann et al., J. Immunol. Methods 1983, vol. 65, p.
55. In all cases the concentrations used were up to 100 mikroM and
times of incubation until 72 h. Table 1 shows the biological
activity (IC.sub.50) of some compounds of formula (I)
TABLE-US-00001 TABLE 1 Biological activities. Compound (I)
IC.sub.50 (.mu.M) Jurkat clon E6-1 IC.sub.50 (.mu.M) GLC-4 Ia 1.42
1.63 Ib 0.59 1.10 Ic 0.95 4.70 Id 3.37 3.57 Ie 5.13 2.88 If 7.16
3.64 Ip 0.26 0.67 Iu 18.7 15.70 Iv Not tested 16.73 Iw Not tested
1.08 Ix 1.21 19.57 Iy 7.94 19.08 Iz 5.02 11.53 Iac Not tested 5.88
Sequence CWU 1
1
318DNAArtificial SequenceSITE 1ttccggaa 8211DNAArtificial
SequenceSITE 2cttcttcttc t 11311DNAArtificial SequenceSITE
3cttcctcctc t 11
* * * * *