Quantitative structure-activity relationships for a series of symmetrical bisquaternary anticancer compounds

Article abstract of DOI:10.1016/S0968-0896(02)00054-8

56 biscationic dibromides with distinct polar heads [bis(4-substituted)pyridinium, bis(4-aminoquinolinium), bisquinolinium, and bisisoquinolinium moieties] and several spacers between the two charged nitrogen atoms were synthesised. This oriented synthesis produced 45 inhibitors of choline kinase with antitumour activity against the HT-29 cell line. In an attempt to understand the antiproliferative activity, a quantitative structure-activity relationship was developed. The unknown σ_R and σ_R⁺ descriptors for the diallylamino, pyrrolidino, piperidino and perhydroazepino groups and σ_R for the N-methylanilino moiety, were estimated by ¹³C NMR spectroscopy in a simple, fast and reproducible manner. The electron characteristic of the substituent at position 4 of the heterocycle and the theoretical lipophilic character of the whole molecule were found to significantly affect the antitumour activity. 1,1′-[Ethylenebis(benzene-1,4-diylmethylene)]bis[4-(N-methylanilino) pyridinium] dibromide is the most active compound of the series so far described and shows a reasonable agreement between predicted and observed antiproliferative data (predicted pIC₅₀=6.50, experimental pIC₅₀=6.46).

Full text of DOI:10.1016/S0968-0896(02)00054-8

Bioorganic & Medicinal Chemistry 10 (2002) 2215–2231
Quantitative Structure–Activity Relationships for a Series of
Symmetrical Bisquaternary Anticancer Compounds
a
a
a
a
Joaquı
´
n M. Campos, Marı
Agustın Rodrıguez-Gonza
Juan Carlos Lacal and A. Espinosa *
´
˜
a C. Nu´ nez, Rosario M. Sanchez, Jose A. Gomez-Vidal,
´ ´ ´
b
b
a
´
´
˜
lez, Monica Banez, Miguel A. Gallo,
´ ´ ´
b
a,
aDepartamento de Quı´mica Farmac e´ utica y Org a´ nica, Facultad de Farmacia, c/Campus de Cartuja s/n, 18071 Granada, Spain
bInstituto de Investigaciones Biom e´ dicas, Consejo Superior de Investigaciones Cientı´ficas, c/Arturo Duperier, 28029 Madrid, Spain
Received 13 December 2001; revised 7 January 2002; accepted 28 January 2002
Abstract—56 biscationic dibromides with distinct polar heads [bis(4-substituted)pyridinium, bis(4-aminoquinolinium), bisquino-
linium, and bisisoquinolinium moieties] and several spacers between the two charged nitrogen atoms were synthesised. This orien-
ted synthesis produced 45 inhibitors of choline kinase with antitumour activity against the HT-29 cell line. In an attempt to
+
R R
understand the antiproliferative activity, a quantitative structure–activity relationship was developed. The unknown s and s
R
descriptors for the diallylamino, pyrrolidino, piperidino and perhydroazepino groups and s for the N-methylanilino moiety, were
estimated by 13C NMR spectroscopy in a simple, fast and reproducible manner. The electron characteristic of the substituent at
position 4 of the heterocycle and the theoretical lipophilic character of the whole molecule were found to significantly affect the
0
antitumour activity. 1,1 -[Ethylenebis(benzene-1,4-diylmethylene)]bis[4-(N-methylanilino)pyridinium] dibromide is the most active
compound of the series so far described and shows a reasonable agreement between predicted and observed antiproliferative data
(
predicted pIC50=6.50, experimental pIC50=6.46). # 2002 Elsevier Science Ltd. All rights reserved.
Introduction
properties, we have applied the classical Hansch analy-
sis to the series of compounds under study lookingfor
correlations between the variation of anti-proliferative
activity and the variation of parameters describingthe
electronic and hydrophobic properties of the mol-
The need to selectively target any drug is not only
desirable but of the utmost necessity, especially in can-
cer chemotherapy. All the anti-neoplastic drugs known
so far are limited to their selectivity and therefore tend
to destroy all actively proliferatingcells, includingthe
normal cells. Medicinal chemists are eager to develop
drug-targeted systems that would lower or eliminate the
side effects of drugs, and thus increase their therapeutic
index.
3
ecules.
Lipid metabolic pathways are frequently altered during
carcinogenesis. Some of them play an important role in
mitogenic signalling such as diacylglycerol and phos-
phatidylinositol. Phosphorylcholine (PCho) is generated
by choline kinase (ChoK) after mitogenic stimulation by
growth factors, and is found increased in human
The advances in quantitative structure–activity rela-
tionship (QSAR) studies have widened the scope of
rationalisingdrugdes ni g and the search for the
mechanisms of drugactions. Bisquaternary salts have
been amongthe favourite classes of chemical com-
4
À6
tumours.
get for the design of antitumour drugs has been repor-
In vivo evidence that ChoK is a novel tar-
7
,8
ted, pointingout that ChoK mi hg t play a role in
growth promotion or signal transduction in carcino-
genesis. We have correlated the inhibitory effect on
1
,2
pounds for QSAR studies. Factors contributingto
such popularity have included the facility of synthesis
and the wide variations in potency induced by changes
in structure. With the aim to rationalize the SAR of
biscationic derivatives in terms of physicochemical
6
proliferation of symmetrical bisquaternary compounds
with their ability to inhibit the production of PCho in
whole cells. When the 1,2-ethylene(bisbenzyl) moiety
was used as a linker between the two 4-substituted
9
pyridinium cationic heads, the structures were screened
for their activity inhibitingisolated ChoK (under ex
vivo conditions). The 4-NR group made a substantial
2
contribution and was suggested that the role of the
*Correspondingauthor. Tel.: +34-958-243850; fax: +34-958-243845;
e-mail: aespinos@ugr.es
9
0
968-0896/02/$ - see front matter # 2002 Elsevier Science Ltd. All rights reserved.
PII: S0968-0896(02)00054-8

2216
J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
4
-NR group was electronic, via delocalisation of the
2
Scheme 2. Compound 64 was previously described by
heatinga mixture of 4-chloropyridine and N-methyl-
positive charge. The importance of frontier orbital
energies (LUMO) of model compounds has been
emphasised and interpreted.
1
1
aniline under reflux on a boilingwater bath. Protona-
tion of the pyridine ring, however, brings about an
increase in the lability of the halogen and, moreover, the
use of a high-boiling-point alcohol led us to an increase
in the yield of 64.
1
0
There is a need for group substituent constants that are
experimental unavailable. Herein, we describe a method
which gives an estimation of several unknown s and
R
+
s
R
descriptors. Although estimated constants are not as
Although the NMR of compounds 1–63 looked clean,
in general there were problems with their elemental
analyses. When the necessary molecules of water were
added, the elemental analyses fitted perfectly. The C, H
and N values did not improve significantly when the
dryingof the samples was carried out in hi gh vacuum at
higher temperatures. Nevertheless, the situation was
dependable as reliable experimental constants, they are
nevertheless of considerable use. We have tried to cor-
relate p(IC )
5
of the whole set of compounds with
0 ChoK
the electronic and lipophilic parameters. The attempts
have been fruitless but when the set of compounds was
smaller, it was suggested the occurrence of either charge
transfer or bipolar interactions.10 The activity against
the HT-29 cell line is normally higher than the corre-
sponding activity against ChoK and it might be possible
that the bisquaternary salts also act on another
unknown target (see Tables 4–6). The substitution pat-
tern of the bis(4-substitutedpyridinium) moiety and the
modification of the spacer connectingthe two cationic
heads have led to a more potent anti-proliferative sym-
metrical bisquaternary compound that arises from a
QSAR study.
1
13
different for compound 23 (Table 1): in the H and
C
NMR spectra, each signal appeared in duplicate, show-
ing, therefore, the presence of a second very similar
structure (23b) to the bis[4-(acetil)pyridinium] com-
1
pound (23a). The H NMR spectrum showed that
compound 23a is in 69% in the mixture. Keepingin
mind that water is present in 23a, it is plausible that it
could be chemically added to the carbonyl groups to
give the bis(gemdiol) form 23b. This might be the only
possibility for the compatibility of the elemental analy-
sis for the two structures 23a and 23b. The p-deficient
heterocyclic system exerts a significant electron-with-
drawingeffect on the carbonyl gr oup and consequently
nucleophilic attack at that centre takes place readily, in
spite of the poor nucleophilicity of water. The situation
is similar to highly halogenated ketones and aldehydes,
which form stable hydrates due to the electronegativity
Results and Discussion
Chemistry
Compounds 1–63 were prepared by quaternisation of
the heterocycle nitrogen by the appropriate dibromide
in butanone as described previously (Scheme 1).9,10 The
species comprisingthe transition state of the reaction
are more polar than the reagents and the lone pair of
the endocyclic nitrogen of the aromatic system will have
to be free to attack the electrophile. Therefore the sol-
vent must be polar to stabilise the transition state and
aprotic to not interact with the N-lone pair by hydro-
gen-bonding. Butanone was then selected.
1
2
of the groups attached to the carbon carbon. Scott’s
group reported a direct application of NMR spectro-
scopy to the structure of well-designed tetrahedral inhi-
1
3
13
bitor complexes. In the case of trypsin,
a
C NMR
study of the complex between the enzyme and p-amidino-
phenylpyruvic acid revealed characteristic resonances (d
202.5 ppm for the keto group of the substrate and d 95.4
ppm for the hydrated keto form). In our case, the reso-
nance of the carbonyl group of 23a was at d 195.56 ppm
and that of the gemdiol carbon of 23b was at d 98.14
ppm. The same hydratation of the carbonyl moieties
was found for the rest of the bis(acetyl)pyridinium
structures (5a,b, 30a,b, 10a,b and 16a,b). Table 1 shows
Compound
obtained by reactingthe 4-chloropyridine hydrochloride
4-(N-methylanilino)pyridine
64 was
and N-methylaniline in refluxingpentanol as shown in
Table 1. Ratio of the bis[4-(acetyl)pyridinium] 5a, 30a, 10a, 16a and 23a and the correspondingbis( gemdiol) compounds 5b, 30b, 10b, 16b and 23b
Space
isomer
Z
Bisacetyl
form/ (%)
Bis(gemdiol)
form/(%)
Bis[4-(acetyl)]
form (%)
Bis(gemdiol)
form (%)
0
0
0
0
0
00
00
00
00
00
3
4
4
4
4
,3
,4
,4
,4
,4
—
—
CH
5a
5b
43
73
44
69
69
57
27
56
31
31
10a
16a
23a
30a
10b
16b
23b
30b
2
(CH
(CH
2
)
)
2
3
2

J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
2217
the ratio of the bis[4-(acetyl)pyridinium] 5a, 30a, 10a,
6a and 23a and the correspondingbis( gemdiol) com-
pounds 5b, 30b, 10b, 16b and 23b.
same structure. On the other hand, different compounds
with pyridinium protons that show two doublets on the
H NMR spectra at room temperature, relax similarly
to the rest of the aromatic protons on the same structure
1
1
(data not shown).
Spectroscopic behaviour of compounds 57–59
This paper reports on an NMR study of 57–59 (Table
1
For viscous liquids, molecular orientations are not ran-
dom, and transfer of energy spin–lattice relaxation is
efficient and longitudinal relaxation time (T ) is small,
1
13
5
One of the most characteristic features of the H NMR
), includingthe full assi gn ed H and C NMR spectra.
1
15
CD OD spectra of 57–59 is that the signals at ꢀd 8.3
and this is why broad signals were obtained for H-2 and
H-3 of the pyridium moiety of 57–59 in the NMR spec-
3
ppm (H-2 ) and ꢀd 6.9 ppm (H-3 ) are very broad
pyr
pyr
1
3
singlets at room temperature and they sharpen and
change to doublets on warming (see Fig. 1 for details).
tra. In the cases in which the relaxation of C nuclei is
produced by a spin–spin relaxation, it is common to
1
4
1
3
13
Regarding the C NMR CD OD spectrum, it must be
3
notice the broadeningof lines in C NMR spectra. For
1
3
14
pointed out that the signals, at d 143.63 ppm (C-2pyr
)
instance, C nuclei linked to N nuclei appear fre-
4
1
and at d 110.26 ppm (C-3pyr) do not appear in the
DEPT experiment, but they appear at higher tempera-
quently as very broad signals.
ꢁ
tures (ca. 77 C in DMSO-d ).
6
The extension of the conjugation through the 4-amino
substituent on the pyridinium ringmust result in an
overall reduced tumblingof the molecule, and the con-
T of pyridinium protons were determined for structures
1
1
6 1
13
57–59 (see Experimental for details). These protons
relax faster than the rest of the aromatic protons on the
sequent reduction of T1. H and C NMR spectra
were studied in DMSO-d gradually increasing the tem-
6
a
4
+
R
Table 2. CSD of several R moieties of symmetrical bispyridinium structures and bibliographical values of descriptors s
, s
P R
and s
R4
Z
Spacer isomer
CSDa
s
b
P
s
R
b
s
R
+b
Compd
0
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
1
2
3
4
5
6
7
8
9
–NMe
–NH
2
—
—
—
—
—
—
—
—
—
—
—
CH
CH
CH
CH
CH
CH
CH
3 ,3
À3.92
À3.62
À0.71
À0.82
0.22
À3.96
À3.67
À0.66
À0.89
0.22
À0.56
À3.97
À3.71
À0.72
À0.88
0.22
À0.71
À0.63
0.04
À0.88
À0.80
À0.07
À0.16
0.20
À0.88
À0.80
À0.07
À0.16
0.20
À1.22
À1.10
À0.15
À0.25
0.06
À1.22
À1.10
À0.15
À0.25
0.06
À0.12
À1.22
À1.10
À0.15
À0.25
0.06
0
2
3 ,3
0
–CH
2
OH
–Me
–COMe
3 ,3
0
c
c
3 ,3
À0.17
0
3 ,3
0.50
0
–NMe
–NH
2
4 ,4
À0.71
À0.63
0.04
0
2
4 ,4
0
–CH
2
OH
–Me
4 ,4
0
4 ,4
À0.17
0
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
–COMe
–CH¼NOH
4 ,4
0.50
0
c
4 ,4
À0.10
À0.71
À0.63
0.04
—
0
–NMe
–NH
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
4 ,4
À0.88
À0.80
À0.07
À0.16
0.20
0
2
4 ,4
0
–CH
2
OH
–Me
4 ,4
0
c
c
c
4 ,4
À0.17
0
–COMe
–CN
4 ,4
0.50
0.65
0
4 ,4
0.96
0.13
—
0.13
0
c
–CH¼NOH
4 ,4
À0.57
À3.98
À3.72
À0.73
À0.89
0.19
À0.10
À0.71
À0.63
0.04
À0.12
À1.22
À1.10
À0.15
À0.25
0.06
0
–NMe
–NH
2
(CH
(CH
(CH
(CH
(CH
(CH
(CH
(CH
(CH
(CH
(CH
(CH
(CH
2
2
2
2
2
2
2
2
2
2
2
2
2
)
)
)
)
)
)
)
)
)
)
)
)
)
4 ,4
À0.88
À0.80
À0.07
À0.16
0.20
0
2
4 ,4
0
–CH
2
OH
–Me
4 ,4
0
4 ,4
À0.17
0
–COMe
–CN
–COOH
4 ,4
0.50
0.65
0
4 ,4
1.01
0.31
0.13
0.11
0.13
—
0
4 ,4
0.41
0
–NMe
–NH
2
4 ,4
À4.03
À3.77
À0.71
À0.88
0.22
À0.71
À0.63
0.04
À0.88
À0.80
À0.07
À0.16
0.20
À1.22
À1.10
À0.15
À0.25
0.06
0
2
4 ,4
0
–CH
2
OH
–Me
4 ,4
0
4 ,4
À0.17
0
–COMe
–CN
4 ,4
0.50
0.65
0
4 ,4
0.97
0.13
0.13
a
CSD=(d13CR4À d13C
H
3
)CD OD; positive values indicate decreased shielding; CSD values are the average of three measurements.
b
+
P R R
s : Hammett constant for para substitution; s : electronic parameter for resonance effect; s : electronic parameter defined for systems where
+
P R R
a+charge is delocalised between substituent and reaction centre via ‘through resonance’; otherwise stated, the s , s and s values have been
taken from ref 24.
c
Ref 25.

2218
J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
perature (see Fig. 1) to diminish the viscosity and to
increase molecular tumbling, making T larger.
represent a measure of the interaction between the sub-
stituent of the aromatic ringand the reaction centre,
attempts are often made to use a variety of physical
methods to obtain linear correlations between the sigma
values and the given set of physical parameters.
Methods of estimation of substituent constants experi-
mental as yet unavailable are required.
1
1
ꢁ
The H NMR spectrum of 58 was run at 57 C and
showed a doublet at d 8.45 ppm (J2,3=7.5 Hz) and
another one, not so well resolved at d 6.91 ppm. When
1
9
ꢁ
the same spectrum was run at 97 C, the doublet corre-
spondingto H-2 pyr appeared slightly shifted at d 8.42
ppm (J2,3=7.5 Hz) and the signal corresponding to H-
The NMR chemical shift serves as a sensitive probe for
the estimation and calculation of the transmission of the
electronic effects of substituents. The Hammett rela-
tionship has also been extended to pyridine as an
empirical method for correlatingreactivity with aro-
3pyr appeared perfectly defined as a doublet centred at d
ꢁ
showed an intermediate behaviour to that shown at 57
and 97 C.
6
.91 ppm (J2,3=7.4 Hz). The spectrum run at 77 C
ꢁ
2
0,21
matic structure.
In the pyridine ring, unlike the
benzene ring, the nuclear nitrogen atom acts as a func-
tional centre. The nitrogen ring polarizes the ring
towards itself, both inductively and mesomerically.
Such an effect is considerably accentuated in the pyri-
dinium ion, where the nitrogen carries a positive
charge.
Pharmacology
Compounds 1–63 were tested usingpurified ChoK from
yeast. Next, the bisquaternary compounds 1–63 were
screened on anti-proliferative assays, usingthe human
tumour-derived cell line HT-29 mainly resistant to
chemotherapy. Recently, the function of the tyrosine
kinase Src has been identified as an essential step for the
tumorigenic activity of HT-29 cells. Given that ChoK
inhibitors drastically impair proliferation of src-trans-
formed marine fibroblasts, HT-29 cells seemed to be a
1
7
The chemical shifts of the N-methyl protons of sub-
stituted pyridinium halides have been correlated with
1
8
2
2
Hammett constants. As part of a programme for the
development of new ChoK inhibitors as anti-
proliferative drugs, our efforts were focused on the
6
good system to use in the screening for ChoK inhibitors
as new anticancer drugs. The Hill equation was fitted to
the data to obtain estimates of the IC . Table 4 shows
6
synthesis of hemicholinium-3 derivatives. Theoretical
and observed substituent induced chemical shifts of the
methylene protons in N-benzylpyridinium salts were
reported in substantial agreement with those expected
5
0
the structures and biological results for compounds 1–
6 arranged in decreasing order of their inhibitory
potencies of isolated ChoK.
5
2
3
from the overall electronic effects of the substituents.
1
3
The C NMR spectra have been determined for a series
of 31 symmetrical bisquaternary compounds, and the
existence of any correlation between such chemical shift
and the Hammett and Hammett-type constants was
investigated. The 2-substituted pyridinium salts were
omitted owingto steric hindrance effects caused by the
presence of bulky groups close to the reactive site.
Hence, only 4-substituted bis-pyridinium compounds
were used in this investigation.
Application of the 13C chemical shift differences to the
determination of the ꢀR and ꢀR constants of several
dialkylamino groups
+
Over the years, considerable interest has been shown in
the application and use of Hammett parameters for
correlation with electronic properties associated with
various substituents. Since the Hammett constants
4
It should be noted that all the substituents (R ) of 1–31
are electron-releasing, electron-neutral or electron-with-
drawing(Table 2). Throu hg out the data there is a
spread of values without clusteringat either end, which
gives validity to the correlations. Donor substituents
caused an upfield displacement of the methylene reso-
nance accordingto their donatingability, while elec-
tron-withdrawing gr oups produced the opposite effect.
Table 2 shows that 4-dimethylamino (1, 6, 12, 19 and
+
R R
Table 3. Calculated s and s values and errors of several amino,
dialkylamino and cyclic dialkylamino groups from correlations 2
and 3
R4
CSDa
s
R
b
s
R
+c
d
d
d
–
–
–
–
–
–
NH
NMe
N(allyl)
2
À3.97Æ0.04
À3.70Æ0.05
À3.74Æ0.02
À3.97Æ0.03
À4.14Æ0.02
À4.02Æ0.01
À0.80
À1.10Æ0.050
À1.22Æ0.053
À1.09Æ0.033
À1.16Æ0.033
À1.21Æ0.029
À1.18Æ0.028
d
2
À0.88Æ0.028
À0.80Æ0.065
À0.85Æ0.063
À0.89Æ0.057
À0.86Æ0.057
2
e
NC
NC
NC
4
5
6
H
H
H
8
26) and 4-amino (2, 7, 13, 20 and 27) groups produce
f
10
g
12
the maximum upfield shift relative to the unsubstituted
parent compounds. The chemical shift difference (CSD)
a
CSD=(d13CR4À d13C
13
H
)CD OD; positive values indicate decreased
shielding; CSD values are the average of three measurements.
3
is measured as the difference between the C chemical
shift of the methylene group bearing the N+ of the
b
s
P
: Hammett constant for para substitution; s
R
: electronic parameter
: electronic parameter defined for systems
where a+charge is delocalised between substituent and reaction centre
4-substituted bispiridinium and the same CH group of
the 4-unsubstituted one in CD OD. The correlation
+
R
2
for resonance effect; s
3
+
R
via ‘through resonance’; otherwise stated, the s
have been taken from ref 24.
P
, s
R
and s
values
equation that results usingthe Hammett constant for
y
para substitution s is eq 1:
P
c
Ref 25.
Ref 24.
Pyrrolidino.
d
e
y
These compounds group into seven different chemical behaviours.
The equations that describe each one are similar to eq 1, and hence the
conclusions. The same reasoningapplies to eqs 2 and 3.
f
Piperidino.
Perhydroazepino.
g

J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
2219
Table 4. Structure, parameter and calculated log P values, and biological results for the compounds
R4
R +R6
5
Spacer isomer
Z
s
a
s
R
+b
clog Pc
À2.13
CAd
AAe
Compd.
X
Y
R
N+
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
–NH
–NH
–H
(CH¼CH)
2H
4 ,4
4 ,4
0
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
00
(CH
)
À0.80
À0.80
0.00
À0.89
À0.16
À0.86
À0.80
À0.88
0.00
À1.10
À1.10
0.00
À1.21
À0.25
À1.18
À1.09
À1.22
0.00
5
6
9
2
20
2.5
0.4
6
0.4
0.55
2
30
1
2
0.6
2.5
4
3
3
3
3
2
3
3
1
3
3
4
4
2
2
4
4
4
4
4
4
2
4
4
1
5
5
2
4
5
8
5
6
5
5
3
1
3
1
7
5
2
2
9
3
2
1
1
1
5
1
1
1
1
2
2
2
2
3
4
5
9
6
7
9
8
9
0
1
6
0
2
3
4
5
6
7
2
2
2
2
2
2
+
0
f
N
N
N
N
N
N
N
N
(CH¼CH)
t
À5.00
+
0
(CH¼CH)
4 ,4
4 ,4
(CH
(CH
(CH
(CH
(CH
(CH
2
2
2
2
2
2
)
3
)
2
)
3
)
2
)
2
)
2
0.08
À0.93
À1.02
0.09
+
g
0
–NC
5
H
10
2H
2H
9.6
+
0
–Me
4 ,4
4 ,4
12.5
15
17
17
17
0
+
h
0
–NC
–N(allyl)
6
H
12
2H
2H
2H
2H
+
0
2
4 ,4
4 ,4
À0.44
À2.83
À3.10
À1.94
À0.06
À1.53
À2.04
À4.67
+
0
–NMe
–H
2
+
0
3 ,3
4 ,4
—
+
i
0
N
CH
–NC
4
H
8
2H
(CH¼CH)
(CH
(CH
(CH
(CH
(CH
2
)
2
)
3
)
4
)
3
)
2
À0.85
À1.16
+
0
N
–H
–NMe
–NMe
–NH
–NMe
–H
2
4 ,4
4 ,4
2
2
2
2
0.00
0.00
20
22
22
23
23
23
25
25
25
30
31
31
31
33
34
35
35
40
50
60
60
60
60
60
65
70
75
90
90
+
0
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
N
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
2
2
2H
2H
À0.88
À0.88
À0.80
À0.88
0.00
À1.22
À1.22
À1.10
À1.22
0.00
+
0
4 ,4
4 ,4
+
0
2
2H
2H
2H
2H
2H
2H
2H
2H
+
0
f
2
4 ,4
4 ,4
(CH¼CH)
c
À3.16
9
+
0
—
À3.25
À1.15
À2.18
30
0.5
15
15
15
NDl
60
70
40
4
100
>50
NDl
10
NDl
1
+
i
0
–NC
4
H
8
4 ,4
4 ,4
(CH
(CH
2
2
)
)
3
À0.85
À1.16
+
0
–H
–NMe
–NH
–NH
–H
–H
–Me
–H
3
0.00
0.00
+
0
f
2
4 ,4
4 ,4
(CH¼CH)
t
À0.88
À0.80
À0.80
0.00
À1.22
À1.10
À1.10
0.00
À3.16
+
0
f
2
(CH¼CH)
c
À5.00
+
0
2
3 ,3
4 ,4
—
(CH
À4.80
À2.97
À3.4
+
0
8
9
5
0
2H
2H
2
)
2
+
0
4 ,4
4 ,4
CH
CH
(CH
—
(CH
—
—
—
CH
—
(CH
CH
(CH
CH
(CH
CH
—
CH
(CH
(CH
—
—
(CH
—
—
—
—
CH
CH
CH
CH
(CH
(CH
(CH
2
0.00
À0.16
0.00
À0.25
+
0
2H
(CH¼CH)
2
À2.25
À0.43
+
0
2
2
4 ,4
3 ,3
2
)
2
0.00
—
0.00
—
+
j
0
k
–COMe
–CH
2H
2H
À4.21
+
0
8
1
2
2
OH
4 ,4
3 ,3
2
)
3
À0.07
À0.16
0.00
À0.07
À0.85
À0.88
0.00
À0.15
À0.25
0.00
À0.15
À1.16
À1.22
0.00
À3.46
À1.95
À0.71
À4.53
À2.37
À3.12
À0.57
À1.00
+
0
N
N
N
N
–Me
–H
2H
(CH¼CH)
+
0
4 ,4
4 ,4
+
0
–CH
–NC
2
OH
i
2H
2H
+
0
4
H
8
4 ,4
4 ,4
2
+
0
N
–NMe
–H
–H
–COMe
–NMe
–CN
2
2H
20
20
20
NDl
5
+
0
3
4
0
2
1
4
CH
CH
N
N
(CH¼CH)
(CH¼CH)
2H
2
2
4 ,4
4 ,4
2
)
2
3
3
+
0
2
0.00
—
0.00
—
+
j
0
k
N
N
N
N
N
N
N
N
N
N
N
N
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
CH
4 ,4
4 ,4
2
)
À3.49
+
0
2
2H
2H
2H
2H
2
À0.88
À1.22
À3.27
À2.36
À4.68
À4.95
À0.86
À3.88
À1.82
À2.10
À4.38
À4.25
À2.96
+
0
l
4 ,4
4 ,4
2
)
0.13
0.13
ND
+
0
NDl
40
–CH
2
OH
2
À0.07
À0.80
0.00
À0.80
À0.16
À0.16
À0.07
À0.07
À0.88
—
À0.15
À1.10
0.00
À1.10
À0.25
À0.25
À0.15
À0.15
À1.22
—
+
0
–NH
–H
–NH
–Me
–Me
2
4 ,4
4 ,4
+
0
5
7
2
(CH¼CH)
2
2
100
100
100
100
100
100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
>100
6.02
7
20
50
+
0
2
2H
2H
4 ,4
4 ,4
2
)
3
+
0
2
)
2
+
0
2H
2H
2H
2H
2H
2H
4 ,4
3 ,3
+
0
–CH
–CH
2
OH
OH
100
100
20
+
0
1
2
4 ,4
3 ,3
2
)
2
+
0
–NMe
–COMe
2
+
j
0
k
l
0
1
6
3
6
7
8
3
4
5
N
4 ,4
4 ,4
À4.27
ND
+
0
NDl
NDl
N
CH
–CH¼NOH
—
0.00
À0.12
À3.54
À0.85
À5.10
+
0
N
–H
–NH
(CH¼CH)
2
4 ,4
4 ,4
0.00
+
0
l
N
N
N
N
N
N
N
CH
CH
CH
CH
CH
CH
CH
2
2H
2H
2
À0.80
À1.10
—
0.13
ND
NDl
+
j
0
k
–COMe
–CN
4 ,4
4 ,4
2
2
2
—
0.13
À4.51
+
0
l
2H
2H
2H
2H
À3.58
À3.69
ND
+
0
NDl
70
–CH¼NOH
4 ,4
4 ,4
—
—
0.13
0.11
À0.12
+
j
0
k
–COMe
–CN
–COOH
2
)
)
)
2
2
2
—
0.13
À4.02
+
0
l
4 ,4
4 ,4
2
2
À3.15
À3.50
ND
NDl
+
0
2H
—
a
CSD=(d13CR4À d13C
H
3
)CD OD; positive values indicate decreased shielding; CSD values are the average of three measurements.
+
b
P R R
s : Hammett constant for para substitution; s : electronic parameter for resonance effect; s : electronic parameter defined for systems where a+charge is
+
delocalised between substituent and reaction centre via ‘through resonance’; otherwise stated, the s
Predicted by usingthe Ghose–Crippen modified atomic contribution system (ATOMIC5 option) of the PALLAS 2.0 programme.
P
, s
R R
and s values have been taken from ref 24.
27
c
26
d
CA: (IC50
)
ChoK (mM): Choline kinase activity was analysed by measuringthe conversion of labelled Cho into PCho in the presence of different concentra-
tions of compounds.
AA: (IC50
e
)HT-29 (mM): The anti-proliferative activity was analysed on HT-29 cultured cells by measuringthe number of cells remainingafter 6 days of
incubation with the compounds, relative to control, non-treated cells. IC50 refers to the concentration at which 50% inhibition of choline kinase or pro-
liferation activities are reached.
f
The PALLAS programne does not differentiate between the cis- and trans-isomers.
Piperidino.
Perhydroazepino.
g
h
i
Pyrrolidino.
Not included in the derivation of eqs 4 and 5 as it does not exist purely in the bis(acetyl) form but as a mixture of bis(acetyl) and bis(1,1-dihydroxyethyl)
j
forms.
It is calculated as a weighted average of the bis(acetyl) and bis(1,1-dihydroxyethyl) forms.
ND, not determined.
k
l

2220
J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
reasonable correlation between the 13C chemical shift
CSD ¼ À1:08 ðÆ0:10Þ þ 3:55 ðÆ0:20Þ sp
ð1Þ
2
and s (r =0.920), which is enhanced when the reso-
P
+
2
2
n=31, r=0.919, s=0.521, F1,29=331.00, p< 0.001
nance effects sR and sR (r =0.976 and r =0.986,
respectively) are used. When only the resonance com-
ponent is considered (s =s +s ), an excellent corre-
P
I
R
where n is the number of compounds, r is the correla-
tion coefficient, and s is the standard deviation between
estimated and actual chemical shifts. The Fisher test is
highly significant here (p <0.001). The numbers in par-
entheses account for the standard error of the regression
coefficients.
lation results (eq 2):
CSD ¼ À0:27 ðÆ0:06Þ þ 4:23 ðÆ0:26Þ sR
ð2Þ
n=29, r=0.988, s=0.292, F1,27=1,100.35, p< 0.001
Although the nomenclature para is not correct for the
pyridine derivatives, it is used to keep the homogeneity
The resonance effects operate primarily through the
electrons of the p-bond system. Since the p-bond system
of the aromatic ringof the pyridinium compounds also
regarding the Hammett parameter s because the sub-
R
+
stituent is formally para in relation to N . There is a
Table 5. Experimental, and theoretical p(IC50
)HT-29 values for compounds 57, 58 and 59 calculated with correlation 5
a
a
HT-29 experim.
b
HT-29 theoret.
Á (deviation)c
Compound
n
p(IC50
)
ChoK
p(IC50
)
p(IC50
)
5
5
5
7
8
9
1
2
3
5.62
5.19
5.82
6.16
6.46
5.89
6.35
6.50
6.68
0.19
0.04
0.79
a
pIC50=ÀlogIC 50, bearingin mind that the hi gh er the value of pIC 50 the more potent is the compound.
b
Calculated from correlation 5.
Á=p(IC50
c
HT-29 experim.
)HT-29 theoret.Àp(IC50) .
Table 6. Structure, parameter and calculated logP values, and biolo gi cal results for the compounds
R4
R +R
5
6
Z
s
R
a
cLog Pb
p(IC50)c
p(IC50)d
Compd.
X
Y
+
e
60
61
62
57
58
40
59
34
36
51
52
54
63
37
N
N
N
N
N
CH
CH
CH
CH
CH
–NC
–N(Allyl)
6
H
12
2H
2H
2H
2H
2H
(CH
(CH
CH
CH
2
)
)
3
3
À0.86
À0.80
À0.86
À0.78
À0.78
0.00
0.59
0.35
À0.35
À0.06
0.37
5.82
5.82
6.00
5.40
5.14
4.70
5.17
5.04
4.82
4.47
4.30
4.22
5.74
4.77
6.52
6.40
5.89
6.15
6.46
5.70
5.89
5.60
6.40
5.40
5.00
4.70
6.52
6.26
+
+
+
+
2
e
2
–NC
H
6 12
2
–N(Me)Ph
–N(Me)Ph
–H
2
(CH
(CH
(CH
(CH
(CH
(CH
2
)
2
3
3
3
2
2
+
CH
+
N
(CH¼CH)
2
2
2
2
2
2
2
)
)
)
)
)
À0.06
N
N
N
N
N
N
N
N
CH
CH
CH
CH
CH
CH
CH
CH
–N(Me)Ph
–H
2H
(CH¼CH)
2H
À0.78
0.88
0.08
+
0.00
+
e
–NC
H
6 12
À0.86
0.00
0.00
0.09
+
–H
–H
–H
(CH¼CH)
(CH¼CH)
(CH¼CH)
2H
2
2
2
À0.43
À0.71
À0.57
À0.42
À0.44
+
—
+
(CH
(CH
(CH
2
)
)
)
2
3
2
0.00
À0.89
+
f
–NC
5
H
10
–N(Allyl)
2
2
+
2
2H
À0.80
a
R
s : electronic parameter for resonance effect. It was calculated from eq 5.
b
26
(ATOMIC5 option) of the PALLAS 2.0 programme.27
Predicted by usingthe Ghose–Crippen modified atomic contribution system
Choline kinase activity.
c
d
The anti-proliferative activity was analyzed on HT-29 cultured cells. IC50 refers to the concentration at which 50% inhibition of choline kinase or
proliferation activities are reached.
e
Perhydroazepino.
Piperidino.
f

J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
2221
Scheme 1.
contribution system26 (ATOMIC5 option) of the PAL-
2
7
LAS 2.0 programme.
pðIC50ÞHT-29¼ 5:36 À 0:96 ðÆ0:10Þ sþ
R
ð4Þ
ð5Þ
þ 0:35 ðÆ0:04Þ clog P
Scheme 2.
n=37, r=0.907, s=0.309, F2,34=78.55, p< 0.001
contains a positively charged nitrogen, the transmission
of the resonance effect to the methylene carbons is
pðIC50ÞHT-29¼ 5:36 À 1:31 ðÆ0:13Þ sþ
R
+
24,25
expected to be through the sR descriptor
(eq 3):
0
:35 ðÆ0:04Þ clog P
CSD ¼ 0:03 ðÆ0:05Þ þ 3:36 ðÆ0:07Þ sRþ
ð3Þ
n=37, r=0.910, s=0.304, F2,34=81.68, p<0.001
n=30, r=0.993, s=0.210, F1,28=2,121.51, p<0.001
where p(IC )
5
that the higher the value of p(IC )
5
=Àlog(IC
)
, bearingin mind
the more potent
0 HT-29
50 HT-29
0 HT-29
The better correlation is eq 3 and this is consistent with
4
is the compound. Both eqs 4 and 5 give good cross-
validated r values (q ) of 0.82 and 0.833, respectively.
2
CV
2
conventional chemical concepts. R is para to the posi-
tively charged ring nitrogen and is in direct conjugation
The quality of the two eqs 4 and 5 is almost identical.
This is largely due to the high collinearity between sR
4
with it. The greater resonance effect of R would
+
obviously cause better delocalisation of the positive
charge and, hence, the CSD would become more nega-
tive (i.e., negative values indicate increased shielding).
and sR [r=0.997 for the groups shown in Tables 2 and
3 (n=11)]. Eq 5 is prefered since it has a slightly better
standard deviation (s) than eq 4. A most significant aspect
of this study is that every data point was included in the
formulation of eqs 4 and 5. Such results are rarely found
and merit special consideration. We find this to be quite
unusual since one usually finds some outliers in QSAR
work which must be omitted to obtain a high correlation.
+
Values for s and sR parameters are available for only
R
a relatively small number of substituents. In this paper,
+
the s and s values are reported for an acyclic dial-
R
R
kylamino group such as the diallylamino moiety and four
cyclic dialkylamino groups, such as the pyrrolidino,
piperidino, and perhidroazepino moieties (see Table 3).
Based on eq 5, one could predict which type of com-
pound would give better activity. At this stage the
knowledge of the parameter s of a group such as the
R
QSAR of the antiproliferative activity against the HT-29
cell line
N-methylanilino one acquires a great significance. The
N-methylanilino moiety is both a strongelectron-
releasing and a highly lipophilic group and, according
The octanol–water partition coefficient, used in its
logarithmic form (log P), is the most widely accepted
measure of lipophilicity. Reproducibility and accuracy
of experimental log P determinations are compromised
for extremely lipophilic and/or hydrophilic compounds
such as the biscationic structures 1–63. Fragmental
methods make it possible to create data banks and to
perform log P calculations by computer.
4
to eq 5, should be an excellent R substituent for a bis-
pyridinium dibromide as a promisingantitumour com-
pound. However, the sR value of the N-methylanilino
group is not available and, accordingly, it has been esti-
mated by usingcorrelation eq 2, obtainingthe following
values: CSD=À3.65Æ0.037; s =À0.78Æ0.073. From
R
+
R
eq 3, s =À1.07Æ0.038 for the N-methylanilino group.
This Hammett-type parameter for such a group has been
already obtained by Popova et al. (s =À1.26) by the
+
R
Correlations 4 and 5 show the anti-proliferative activ-
ities of bisquaternary compounds, the Hammett-type
rate of amine acylation with p-nitrobenzenesulfonyl
ꢁ
+
28
constants s and sR of the substitutents at position 4
bromide in a nitrobenzene solution at 25 C.
R
of the heterocycle and the clog P values of the bissalts.
Such clog P values reported in Tables 4 and 6 were cal-
culated by usingthe Ghose–Crippen modified atomic
If the p(IC ) values were calculated with corre-
0 HT-29
lation (eq 5) for compounds 57, 58 and 59 and compare
5

2222
J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
1
À1
ꢁ
Figure 1. Expansion of the 400 H MHz spectrum of 58 (26.7 mgmL ) showingthe si gn als for aromatic protons: (a) CD
ꢁ ꢁ ꢁ
6 6 6
d at 57 C; (c) DMSO-d at 77 C; (d) DMSO-d at 97 C.
3
OD at 25 C; (b) DMSO-

J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
2223
them with the experimental ones, compound 59 deviates
Table 5) more than the standard deviation (s) of eq 5. It
is possible that, between a more ample range of clog P,
compound, is expected to provide impetus for develop-
inga new treatment in the field of cancer therapy. Fur-
ther studies with this compounds are underway and will
be published elsewhere.
(
there exists a parabolic relationship between p(IC )
5
0 HT-
and clog P and, hence, the maximum activity is
9
2
reached with compound 58, a decrease of activity taking
place as the number of methylene units increases from 3
onwards [Z=(CH ) , general formula of Table 5]. The
Experimental
Chemistry
2
n
linear equations should be considered as limited seg-
ments of the more general parabolic equation and the
former may be interpreted as indicatinga situation
where the maximum activity had not been reached.
Whenever a small range of clog P values is available and
Meltingpoints (mp) were taken in open capillaries on
an electrothermal meltingpoint apparatus and are
uncorrected. Nuclear magnetic resonance (NMR) spec-
2
1
the addition of (clog P) term is not statistically justifi-
able, a linear equation may give the best correlation and
this could be our case.
tra were recorded on a 400.13 MHz H and 100.03 MHz
1
3
1
C NMR Bruker ARX 400 or 300.13 MHz H and
1
3
75.78 MHz C NMR Bruker AMX-300 spectrometers,
and chemical shifts (ppm) are reported relative to the
solvent peak (CD HOD in CD OD at d 3.31 and 49.9
In order to check this hypothesis compounds, with their
clogP taken in between the values À0.71 and 0.88 were
selected and prepared (Table 6) and correlation eq 6 was
obtained:
2
3
ppm; DMSO in DMSO-d at d 2.50 and 39.5 ppm).
6
CD OD is used as a NMR solvent unless otherwise
3
stated. Signals are designated as follows: s, singlet; d,
doublet; dd, doublet of doublet; ddd, double doublet of
doublet; t, triplet; dt, double triplet; q, quadruplet; m,
multiplet. Longitudinal relaxation times (T ) were
1
determined usingan inversion recovery experiment on
an Inova 400 MHz NMR instrument. T experiments
2
pðIC50Þ
¼ 5:58 À 0:91ðÆ0:31Þðclog PÞ þ
HT-29
0
:34 ðÆ0:16Þclog P À 1:04 ðÆ0:18Þ sR
ð6Þ
1
were conducted in undegassed CD OD solvent. All final
3
n=14, r=0.934, s=0.238, F3,10=23.84, p<0.001
products had satisfactory (withinÆ0.4%) C, H, and N
analyses. High resolution liquid secondary ion mass
spectra (HR LSIMS) were carried out on a VG Auto-
Spec Q high resolution mass spectrometer (Fisons
Instruments). All compounds were dried at 40 C and
0.1 mmHgfor 15 h, but many held on tenaciously to
solvent molecules, especially water which appears to be
Eq 6 is a significant parabolic equation obtained for the
biscationic compounds [F3,10=23.84; (F =12.55,
and satisfied the F test at the level of 99.9%] which gives
ꢁ
)
3,10 0.001
an ideal lipophilic character (clog P ) of 0.18 for maxi-
0
mum activity.
0
bromomethyl)biphenyl,
29
0
bis[p-(bromomethyl)diphe-
a
solvate. 3,3 -Bis(bromomethyl)biphenyl,
3
4,4 -bis
0,31
(
nyl]methane,
3
2
32
Regarding the molecules 57, 58 and 59, the deviations
found between the theoretical values, calculated from
correlation eq 6, and the experimental ones of
bis-p-(bromomethyl)bibenzyl,
2
1,3-
4-bis[4-(bromo-
3
bis[4-(bromomethyl)phenyl]propane,
3
2
0
10
methyl)phenyl]butane, trans- and cis-4,4 -bis(bromo
3
3
p(IC )
5
were: 0.21 (57), À0.07 (58) and 0.09 (59).
methyl)stilbene, 4-(diallylamino)pyridine, 4-piperi-
10
0 HT-29
1
0
10
dinopyridine,
10
4-(perhydroazepino)pyridine,
33,34
19,
0, 21, 22,10 24, 24, 25, 32,
1
0
10
10
10
36, 37,10 39,
10
10
2
1
0
and 48, were synthesized accordingto literature pro-
cedures. Quinoline, isoquinoline, pyridine, 4-aminopyri-
dine, 4-(dimethylamino)pyridine, 4-pyrrolidinopyridine,
4-(hydroxymethyl)pyridine, 4-methylpyridine and 4-ace-
tylpyridine were obtained from Aldrich.
Conclusions
The electronic and lipophilic parameters were found to
play important roles in the antitumour activity of bis-
1
3
quaternary dibromides. A C NMR method was inclu-
ded which can complement the classic procedure for
determination of these constants for several dialkyl-
amino groups. It has been shown that there is a
remarkable consistency between the results obtained
with this method usingproper assumptions. Within the
range of the data, reasonable prediction should be pos-
sible by eq 5, which predicts approximately 82.6% of
the variance of the p(IC50)HT-29. The present study sheds
light on the influence of the electronic properties of
groups placed at position 4 of the heterocycle and on
the global lipophilicity of compounds. Strongly elec-
tron-donating and highly lipophilic groups provide
analogues with interesting antiproliferative properties.
The 4-(N-methylanilino) group appears to be particu-
larly effective in providinga hi gh ly active compound.
The discovery of compound 58, as a novel anticancer
.
64 HCl.
4-(N-Methylanilino)pyridine
hydrochloride
.
Three grams (19.9 mmol) of 4-chloropyridine HCl and
6.43 mL (59.9 mmol) of N-methylaniline were dissolved
in n-pentanol (40 mL). Such a solution was kept at
reflux for 24 h under argon and then was rotaevapo-
rated off. A pale yellow oil was isolated by flash chro-
matography using a gradient elution (CH Cl !CH Cl /
2
2
2
2
MeOH: 100/5!CH Cl /MeOH: 100/10), which after
2
2
dissolvingin diethyl ether and addingsome drops of a
saturated solution of hydrogen chloride in diethyl ether
precipitated. This white solid was identified as the title
ꢁ
compound (2.89 g, 79%). Mp 189–191 C. TLC (top
1
layer of n-BuOH/HOAc/H O: 5/1/4): 0.38. H NMR
2
(400.13 MHz): d 8.16 (d, J2,3=7.5 Hz, 2H, pyridinium-
H ), 7.60 (t, J =7.6 Hz, 2H, anilino-H ), 7.49 (t, J=7.4
4
3,4
3

2224
J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
Hz, 1H, anilino-H ), 7.37 (d, J2,3=7.4 Hz, 2H, anilino-
4
calcd C, 55.22; H, 4.78; N, 4.95. Found: C, 55.28; H,
5.09; N, 4.83.
H ), 6.90 (d, J =7.5 Hz, 2H, pyridinium-H ), 3.54 (s,
3
2
2,3
1
3
6
1
1
4
H, CH N). C NMR (100.03 MHz): d 159.49 (C-4pyr),
3
0
nium) dibromide (4). Yield: 33.3%. Mp 142–144 C dec.
0
44.97 (C-1anilino), 140.74 (C-2pyr), 131.99 (C-3anilino),
29.98 (C-4anilino), 127.60 (C-2anilino), 109.42 (C-3pyr),
1,1 -[Biphenyl-3,3 -diylbis(methylene)]bis(4-methylpyridi-
ꢁ
+
1
1.24 (CH N). HR LSIMS (thioglycerol+Na ), calcd
3
H NMR (400.13 MHz): d 9.02 (d, J=6.6 Hz, 4H, H-
2pyr), 8.01 (s, 2H, Ph), 7.95 (d, J=6.6 Hz, 4H, H-3pyr),
+
m/z for C H N NaCl (M+Na) 243.0665. Found m/
1
.
z: 243.0664. Anal. for C H N Cl H O: calcd: C, 57.21;
2
13
2
7.76 (d, J=7.5 Hz, 2H, Ph), 7.54 (m, 4H, Ph), 5.88 (s,
+
1
2
13
2
2
13
H, 4.26; N, 3.70. Found: C, 57.07; H, 4.44; N, 3.76. The
free base 64 was liberated from 4-(N-methylanilino)-
4H, CH N ), 2.67 (s, 6H, CH3).
2
C
NMR
(100.13 MHz): d 161.90 (C-4pyr), 144.95 (C-2pyr), 142.61
.
.
pyridine HCl 64 HCl after treatment with aqueous
sodium hydroxide and subsequent extraction with die-
thyl ether.
(C-1 ), 135.68 (C-3 ), 131.36 (C-5 ), 129.54, 129.45,
+
Ph
Ph
Ph
129.19 (C-4,6,2 ), 126.14 (C-3 ), 64.71 (CH N ),
Ph
pyr
2
.
22.06 (Me). Anal. for C H N Br H O: calcd C,
2
6
26
2
2
2
5
4
7.37; H, 5.18; N, 5.15. Found: C, 57.30; H, 5.12; N,
.79.
General experimental procedure for the preparation of
bisquaternary compounds
0 0
,1 -[Biphenyl-3,3 -diylbis(methylene)]bis(4-acetylpyridi-
nium) dibromide (5a). Yield: 76% (mixture of 5a and
1
The heterocyclic structure (1.10 mmol) and the corre-
spondingbis(bromomethyl) compound BrCH ZCH Br
ꢁ
ꢁ
5b). Mp 166–168 C, 148–150 C dec (mixture of 5a and
2
2
ꢁ
1
(
0.54 mmol) in dry butanone (50 mL) were heated in a
5b); 166–168 C liquifies . H NMR (400.13 MHz): d 9.42
(d, J=6.6 Hz, 4H, H-2pyr), 8.51 (d, J=6.6 Hz, 4H, H-
3pyr), 8.11 (s, 2H, Ph), 7.79 (m, 6H, Ph), 6.01 (s, 4H,
ꢁ
sealed tube at 100 C for 24 h. After filtration and
washingthorou hg ly with butanone and CHCl 3, the
solid product was purified by recrystallisation from
+
13
CH N ), 2.75 (s, 6H, CH CO).
2
C
NMR
3
EtOH or EtOH/MeOH, after addingEt O to turbidity.
2
(100.13 MHz, selected data): d 195.50 (CH CO), 65.75
3
+
(
CH N ), 29.06 CH CO).
2 3
0
no)pyridinium] dibromide (1). Yield: 66.3%. Mp 245–
0
1
,1 -[Biphenyl-3,3 -diylbis(methylene)]bis[(4-dimethylami-
0
0
droxy-ethyl)]acetylpyridinium} dibromide (5b). H NMR
1,1 -[Biphenyl-3,3 -diylbis(methylene)]bis{[4- (1,1-dihy-
1
ꢁ
1
2
4
47 C. H NMR (300.13 MHz): d 8.36 (d, J=7.9 Hz,
H, H-2pyr), 7.84 (s, 2H, Ph), 7.69 (d, J=7.7 Hz, 2H,
(400.13 MHz): d 9.20 (dd, J=6.8 and 1.7 Hz, 4H, H-
2pyr), 8.20 (d, J=6.6 Hz, 4H, H-3pyr), 8.11 (d, J=1.4
Hz, 2H, Ph), 7.78 (m, 2H, Ph), 7.58 (m, 4H, Ph), 5.96 (s,
Ph), 7.52 (t, J=7.7 Hz, 2H, Ph), 7.41 (d, J=7.7 Hz, 2H,
Ph), 7.02 (d, J=7.9 Hz, 4H, H-3pyr), 5.48 (s, 4H,
+
13
+
13
CH N ), 3.24 (s, 12H, NMe ). C NMR (75.78 MHz):
2
4H, CH N ), 1.60 (s, 6H, CH C(OH) ). C NMR
2 3 2
2
d 157.99 (C-4pyr), 143.17 (C-2pyr), 142.58 (C-1 ), 137.04
(100.13 MHz, selected data): d 165.74 (CH C(OH) ),
65.06 (CH N ), 27.11 CH C(OH) ). Anal. for
2 3 2
C H N O Br (mixture of 5a and 5b): calcd C, 57.74;
28 26 2 2 2
H, 4.50; N, 4.81. Found: C, 58.06; H, 4.85; N, 4.47.
Ph
3
2
+
(
C-3 ), 131.10 (C-5 ), 128.89, 128.74, 128.56 (C-
+
Ph
Ph
4
HR LSIMS (thioglycerol), calcd m/z for C H N Br
,6,2 ), 109.21 (C-3 ), 61.61 (CH N ), 40.42 (NMe ).
Ph
pyr
2
2
2
8
32
4
+
(
MÀBr) 503.1810. Found m/z: 503.1809. Anal. for
.
28 32 4 2 2
0
0
no)pyridinium dibromide (6). Yield: 79%. Mp 313–
C H N Br 2.81H O: calcd C, 52.96; H, 5.97; N, 8.82.
1,1 -[Biphenyl-4,4 -diylbis(methylene)]bis[(4-dimethylami-
Found: C, 53.35; H, 5.64; N, 8.43.
ꢁ
4H, H-2pyr), 7.69 (d, J=8.4 Hz, 4H, Ph), 7.49 (d, J=8.4
1
3
15 C. H NMR (300.13 MHz): d 8.28 (d, J=7.9 Hz,
0
0
nium) dibromide (2). Yield: 70%. Mp: 310–312 C dec.
1
,1 -[Biphenyl-3,3 -diylbis(methylene)]bis(4-aminopyridi-
ꢁ
Hz, 4H, Ph), 7.02 (d, J=7.9 Hz, 4H, H-3 ), 5.43 (s,
pyr
13
1
+
H NMR (300.13 MHz): d 8.26 (d, J=7.6 Hz, 4H, H-
2pyr), 7.78 (s, 2H, Ph), 7.69 (d, J=7.8 Hz, 2H, Ph), 7.53
4H, CH N ), 3.25 (s, 12H, NMe ).
2
C NMR
2
(75.78 MHz): d 158.02 (C-4pyr), 143.15 (C-2pyr), 142.00
(
(
t, J=7.8 Hz, 2H, Ph), 7.38 (d, J=7.8 Hz, 2H, Ph), 6.87
+
(C-1 ), 135.70 (C-4 ), 130.07 (C-3 ), 128.93 (C-2 ),
+
Ph
Ph
Ph
Ph
13
d, J=7.6 Hz, 4H, H-3pyr), 5.42 (s, 4H, CH N ).
2
C
109.19 (C-3pyr), 61.36 (CH N ), 40.41 (NMe ). HR
2 2
NMR (75.78 MHz): d 160.90 (C-4pyr), 144.13 (C-2pyr),
LSIMS (thioglycerol), calcd m/z for C H N Br (MÀ
28
32
4
+
1
42.68 (C-1 ), 136.97 (C-3 ), 131.14 (C-5 ), 128.95,
Br) 503.1810. Found m/z: 503.1811. Anal. for
C H N Br 2.2H O: calcd C, 53.89; H, 5.88; N, 8.98.
Found: C, 54.17; H, 5.71; N, 8.63.
Ph
Ph
Ph
.
1
(
28.62, 128.35 (C-4,6,2Ph), 111.03 (C-3pyr), 61.91
+
28
32
4
2
2
.
CH N . Anal. for C H N Br 0.1H O: calcd C,
2
24 24
4
2
2
5
1
4.38; H, 4.60; N, 10.57. Found: C, 54.38; H, 4.56; N,
0.75.
0
0
1,1 -[Biphenyl-4,4 -diylbis(methylene)]bis(4-aminopyridi-
ꢁ
1
nium) dibromide (7). Yield: 63.3%. Mp >310 C. H
NMR (400.13 MHz): d 8.20 (d, J=7.4 Hz, 4H, H-2pyr),
7.71 (d, J=8.2 Hz, 4H, Ph), 7.47 (d, J=8.2 Hz, 4H, Ph),
6.87 (d, J=7.4 Hz, 4H, H-3 ), 5.39 (s, 4H, CH N ).
2
C NMR (100.13 MHz): d 160.92 (C-4pyr), 144.11 (C-
2pyr), 142.30 (C-1 ), 135.68 (C-4 ), 129.96 (C-3 ),
0 0
,1 -[Biphenyl-3,3 -diylbis(methylene)]bis[(4-hydroxyme-
thyl)pyridinium] dibromide (3). Yield: 69%. Mp 76–
1
ꢁ
1
+
7
4
8 C. H NMR (400.13 MHz): d 9.11 (d, J=6.6 Hz,
H, H-2pyr), 8.08 (d, J=6.6 Hz, 4H, H-3pyr), 8.02 (s, 2H,
Ph), 7.77 (d, J=7.1 Hz, 2H, Ph), 7.55 (m, 4H, Ph), 5.92
pyr
1
3
Ph
Ph
Ph
+
13
+
(
s, 4H, CH N ), 4.91 (s, 4H, CH OH). C NMR
2
128.98 (C-2 ), 111.00 (C-3 ), 61.62 (CH N ). HR
Ph pyr 2
2
(
100.13 MHz): d 165.35 (C-4pyr), 145.35 (C-2pyr), 142.59
LSIMS (thioglycerol), calcd m/z for C H NBr
24 24
+
(
1
C-1 ), 135.70 (C-3 ), 131.37 (C-5 ), 129.54, 129.49,
+
(MÀBr) 447.1184. Found m/z: 447.1185. Anal. for
Ph
Ph
Ph
.
C H N Br 0.5H O: calcd C, 53.65; H, 4.69; N, 10.63.
Found: C, 53.77; H, 4.55; N, 10.18.
29.20 (C-4,6,2 ), 126.13 (C-3 ), 64.82 (CH N ),
Ph
pyr
2
24 24
4
2
2
.
2.87 (CH OH). Anal. for C H N O Br 0.4H O:
6
2
26 26
2
2
2
2

J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
2225
0
thyl)pyridinium] dibromide (8). Yield: 62%. Mp
0
0
methylamino)pyridinium] dibromide (12). Yield: 72%.
1
,1 -[Biphenyl-4,4 -diylbis(methylene)]bis[(4-hydroxyme-
1,1 -[Methylenebis(benzene-1,4-diylmethylene)]bis[(4-di-
ꢁ
1
ꢁ
1
>
315 C. H NMR (400.13 MHz): d 9.02 (d, J=6.5 Hz,
H, H-2pyr), 8.09 (d, J=6.5 Hz, 4H, H-3pyr), 7.75 (d,
J=8.2 Hz, 4H, Ph), 7.62 (d, J=8.2 Hz, 4H, Ph), 5.88 (s,
Mp 190–192 C. H NMR (300.13 MHz): d 8.21 (d,
J=7.9 Hz, 4H, H-2pyr), 7.31 (d, J=8.3 Hz, 4H, Ph),
7.25 (d, J=8.3 Hz, 4H, Ph), 6.99 (d, J=7.9 Hz, 4H, H-
4
+
13
+
4
H, CH N ), 4.93 (s, 4H, CH OH).
2
C NMR
3pyr), 5.33 (s, 4H, CH N ), 3.98 (s, 2H, CH2 Ph), 3.24 (s,
2
2
1
3
(
100.13 MHz): d 165.45 (C-4pyr), 145.33 (C-2pyr), 142.76
12H, Me). C NMR (75.78 MHz): d 157.96 (C-4pyr),
(
C-1 ), 134.43 (C-4 ), 130.77 (C-3 ), 129.32 (C-2 ),
+
143.58 (C-1 ), 143.03 (C-2 ), 134.06 (C-4 ), 130.84
(C-3 ), 129.63 (C-2 ), 109.11 (C-3pyr), 61.44
Ph Ph
Ph
Ph
Ph
Ph
Ph
pyr
Ph
1
LSIMS (thioglycerol), calcd m/z for C H N O
(MÀBrÀBrH) 397.1916. Found m/z: 397.1916. Anal.
for C H N O Br : calcd C, 55.93; H, 4.69; N, 5.02.
26.15 (C-3pyr), 64.65 (CH N ), 62.88 (CH OH). HR
2
2
+
(CH N ), 41.97 (CH2Ph), 40.38 (Me). HR LSIMS
2
2
6
25
2
2
+
+
(thioglycerol), calcd m/z for C H N Br (MÀBr)
2
9
34
4
517.1967.
Found
m/z:
517.1966.
Anal.
for
2
6
26
2
2
2
.
C H N Br 2.5H O: calcd C, 54.13; H, 6.11; N, 8.71.
29 34 4 2 2
Found: C, 55.73; H, 4.72; N, 5.39.
Found: C, 54.10; H, 5.96; N, 8.40.
0 0
,1 -[Biphenyl-4,4 -diylbis(methylene)]bis(4-methylpyridi-
nium) dibromide (9). Yield: 87%. Mp 303–305 C. H
NMR (300.13 MHz): d 8.94 (d, J=6.6 Hz, 4H, H-2pyr),
1
ꢁ
1
0
nopyridinium) dibromide (13). Yield: 79%. Mp 254–
1,1 -[Methylenebis(benzene-1,4-diylmethylene)]bis(4-ami-
ꢁ
1
7
Ph), 7.61 (d, J=8.4 Hz, 4H, Ph), 5.85 (s, 4H, CH N ),
.96 (d, J=6.6 Hz, 4H, H-3pyr), 7.73 (d, J=8.4 Hz, 4H,
+
256 C. H NMR (300.13 MHz): d 8.14 (d, J=7.6 Hz,
4H, H-2pyr), 7.30 (d, J=8.3 Hz, 4H, Ph), 7.25 (d,
J=8.3, 4H, Ph), 6.85 (d, J=7.6 Hz, 4H, H-3pyr), 5.30 (s,
2
1
3
2.68 (s, 6H, CH3). C NMR (75.78 MHz): d 161.89 (C-
4pyr), 144.93 (C-2pyr), 142.68 (C-1 ), 134.46 (C-4 ),
+
13
4H, CH N ), 3.97 (s, 2H, CH2Ph).
2
C NMR
(75.78 MHz): d 160.78 (C-4pyr), 143.99 (C-2pyr), 143.56
Ph
Ph
1
30.78 (C-3 ), 130.17 (C-2 ), 129.18 (C-3 ), 64.43
Ph
pyr
Ph
+
(
CH N ), 22.06 (Me). HR LSIMS (thioglycerol), calcd
2
(C-1 ), 134.04 (C-4 ), 130.88 (C-3 ), 129.61 (C-2 ),
Ph
Ph
Ph
Ph
+
+
m/z for C H N Br (MÀBr) 445.1279. Found m/z:
110.96 (C-3pyr), 61.70 (CH N ), 41.98 (CH2Ph). HR
2
2
6
26
2
.
45.1279. Anal. for C H N Br 0.8H O: calcd C,
7.75; H, 5.15; N, 5.18. Found: C, 57.57; H, 4.84; N,
.91.
4
5
4
LSIMS (thioglycerol), calcd m/z for C H N Br
4
2
6
26
2
2
2
25 26
+
(MÀBr) 461.1341. Found m/z: 461.1339. Anal. for
.
25 26 4 2 2
C H N Br H O: calcd C, 53.59; H, 5.04; N, 10.00.
Found: C, 53.49; H, 4.96; N, 9.76.
0 0
,1 -[Biphenyl-4,4 -diylbis(methylene)]bis(4-acetylpyridi-
nium) dibromide (10a). Yield: 63% (mixture of 10a and
1
0
droxymethyl)pyridinium] dibromide (14). Yield: 71%.
1,1 -[Methylenebis(benzene-1,4-diylmethylene)]bis[(4-hy-
ꢁ
1
1
0b). Mp >315 C dec (mixture of 10a and 10b). H
NMR (400.13 MHz): d 9.33 (d, J=6.6 Hz, 4H, H-2pyr),
.52 (d, J=6.6 Hz, 4H, H-3pyr), 7.76 (d, J=8.3 Hz, 4H,
ꢁ
ꢁ
1
Mp 199–200 C, 153–155 C dec.
H
NMR
8
(300.13 MHz): d 8.97 (d, J=6.7 Hz, 4H, H-2pyr), 8.05
(d, J=6.7 Hz, 4H, H-3pyr), 7.45 (d, J=8.1 Hz, 4H, Ph),
7.30 (d, J=8.1 Hz, 4H, Ph), 5.79 (s, 4H, CH N ), 4.90
+
Ph), 7.69 (d, J=8.3 Hz, 4H, Ph), 6.02 (s, 4H, CH N ),
2
2
1
3
+
.76 (s, 6H, CH CO). C NMR (100.13 MHz, selected
3
2
13
+
data): d 195.55 (CH CO), 65.49 (CH N ), 29.15 (CH ).
(s, 4H, CH OH), 4.00 (s, 2H, CH Ph). C NMR
3
2
3
2
2
(
75.78 MHz): d 165.21 (C-4pyr), 145.17 (C-2pyr), 144.21
0 0
,1 - [Biphenyl -4,4 -diylbis(methylene)]bis{[4 -(1,1- dihy-
1
(C-1 ), 132.80 (C-4 ), 131.10 (C-3 ), 130.40 (C-2 ),
+
Ph
Ph
Ph
Ph
1
droxy-ethyl)]acetylpyridinium} dibromide (10b).
NMR (400.13 MHz): d 9.10 (d, J=6.6 Hz, 4H, H-2pyr),
.21 (d, J=6.6 Hz, 4H, H-3pyr), 7.76 (d, J=8.3 Hz, 4H,
H
126.07 (C-3 ), 64.69 (CH N ), 62.86 (CH OH), 41.99
pir 2 2
(CH2Ph). HR LSIMS (thioglycerol), calcd m/z for
+
8
C H N O Br (MÀBr)
491.1334. Found m/z:
27
28
2
2
+
.
491.1335. Anal. for C H N O Br 0.2H O: calcd C,
27 28 2 2 2 2
Ph), 7.68 (d, J=8.3 Hz, 4H, Ph), 5.93 (s, 4H, CH N ),
2
1
3
1
.62 (s, 6H, CH C(OH) ). C NMR (100.13 MHz,
3
56.31; H, 4.97; N, 4.86. Found: C, 56.20; H, 4.99; N, 5.08.
2
+
selected data): d 98.14 (CH C(OH) ), 64.81 (CH N ),
3
2
2
0
thylpyridinium) dibromide (15). 95 C. Mp 232–235 C.
2
mixture of 10a and 10b, calcd m/z for C H N O Br
7.12 (CH C(OH) ). HR LSIMS (thioglycerol) for the
1,1 -[Methylenebis(benzene-1,4-diylmethylene)]bis(4-me-
ꢁ
3
2
ꢁ
2
8
26
2
2
+
1
(MÀ Br) 501.1178. Found m/z: 501.1178. Anal. for
C H N O Br 1.5H O (mixture of 10a and 10b): calcd
H NMR (300.13 MHz): d 8.87 (d, J=6.6 Hz, 4H, H-
2pyr), 7.93 (d, J=6.6 Hz, 4H, H-3pyr), 7.43 (d, J=8.2
.
2
8
26
2
2
2
2
C, 55.19; H, 4.80; N, 4.60. Found: C, 55.28; H, 4.89; N,
4
Hz, 4H, Ph), 7.30 (d, J=8.2 Hz, 4H, Ph), 5.75 (s, 4H,
13
+
.87.
CH N ), 4.00 (s, 2H, CH2 Ph), 2.66 (s, 6H, Me).
2
C
NMR (75.78 MHz): d 161.75 (C-4pyr), 144.79 (C-2pyr),
0
0
1,1 -[Biphenyl-4,4 -diylbis(methylene)]bis[(4-hydroxyimi-
nomethyl)pyridinium] dibromide (11). Yield: 59%. Mp
2
144.19 (C-1 ), 132.82 (C-4 ), 131.09 (C-3 ), 130.35
(C-3pyr or C-2 ), 130.10 (C-2
Ph
Ph
Ph
or C-3pyr), 64.53
Ph
Ph
ꢁ
1
+
44–246 C. H NMR (400.13 MHz): d 9.04 (d, J=6.8
Hz, 4H, H-2pyr), 8.32 (s, 2H, CH), 8.24 (d, J=6.8 Hz,
H, H-3pyr), 7.75 (d, J=8.3 Hz, 4H, Ph), 7.64 (d, J=8.3
(CH N ), 41.98 (CH2Ph), 22.03 (Me). HR LSIMS
2
+
(thioglycerol), calcd m/z for C H N (MÀBrÀBrH)
27
27
2
4
379.2174.
Found
m/z:
379.2173.
Anal.
for
+
13
.
Hz, 4H, Ph), 5.88 (s, 4H, CH N ).
2
C NMR
100.13 MHz): d 151.65 (C-4pyr), 146.01 (C-2pyr or C-
C H N Br 0.3H O: calcd C, 59.42; H, 5.28; N, 5.13.
27 28 2 2
2
(
5Ph), 145.59 (C-5Ph or C-2pyr), 142.75 (C-1 ), 134.26 (C-
Found: C, 59.36; H, 5.55; N, 4.99.
Ph
0
ylpyridinium) dibromide (16a). Yield: 38% (mixture of
4Ph), 130.88 (C-3 ), 129.24 (C-2 ), 125.70 (C-3pyr),
Ph
.
4.76 (CH N ). Anal. for C H N O Br 0.4H O:
calcd C, 52.79; H, 4.23; N, 9.47. Found: C, 52.72; H,
.01; N, 9.18.
1,1 -[Methylenebis(benzene-1,4-diylmethylene)]bis(4-acet-
Ph
+
6
2
26 24
4
2
2
2
ꢁ
16a and 16b). Mp 185–187 C dec (mixture of 16a and
1
4
16b). H NMR (400.13 MHz): d 9.26 (d, J=6.9 Hz, 4H,

2226
J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
+
H-2pyr), 8.48 (d, J=6.9 Hz, 4H, H-3pyr), 7.50 (d, J=8.1
Hz, 4H, Ph), 7.32 (d, J=8.1 Hz, 4H, Ph), 5.91 (s, 4H,
7.30 (d, J=8.3 Hz, 4H, Ph), 5.82 (s, 4H, CH N ), 2.94
2
1
3
(s, 4H, CH Ph), 1.52 (s, 6H, CH C(OH) ). C NMR
2
(75.78 MHz): d 98.14 (CH C(OH) ), 65.02 (CH N ),
27.13 (CH C(OH) ). HR LSIMS (thioglycerol) for the
mixture of 23a and 23b, calcd m/z for C H N O Br
30 30 2 2
3
2
+
+
CH N ), 4.02 (s, 2H, CH Ph), 2.74 (s, 6H, CH CO).
2
2
3
3
2
2
1
3
C NMR (100.623 MHz, selected data): d 195.55
3 2
+
(
CH CO), 65.62 (CH N ), 29.11 (CH ).
3
2
3
+
(
MÀBr) 529.1491. Found m/z: 529.1593. Anal. for
.
30 30 2 2 2 2
0
1,1-dihydroxy-ethyl)]acetylpyridinium} dibromide (16b).
1
,1 - [Methylenebis(benzene - 1,4 - diylmethylene)]bis{[4 -
C H N O Br 2H O (mixture of 23a and 23b): calcd
(
C, 55.74; H, 5.30; N, 4.33. Found: C, 55.87; H, 5.61; N,
4.39.
1
H NMR (400.13 MHz): d 9.03 (d, J=6.9 Hz, 4H, pyr-
idine-H ), 8.17 (d, J=6.9 Hz, 4H, pyridine-H ), 7.47 (d,
J=8.1 Hz, 4H, Ph), 7.32 (d, J=8.1 Hz, 4H, Ph), 5.82 (s,
2
3
0
bis[(4-dimethylamino)pyridinium) dibromide (26). Yield:
1,1 - [Propane - 1,3 - diylbis(benzene - 1,4 - diylmethylene)]-
+
4
H, CH N ), 4.02 (s, 4H, CH Ph), 1.60 (s, 6H,
2
2
1
3
ꢁ
1
CH C(OH) ). C NMR (100.03 MHz, selected data): d
92%. Mp 133–135 C. H NMR (300.13 MHz): d 8.24
(d, J=7.8 Hz, 4H, H-2pyr), 7.32 (d, J=8.1 Hz, 4H, H-
3pyr), 7.24 (d, J=8.1, 4H, Ph), 7.00 (d, J=7.8 Hz, 4H,
3
2
+
9
8.13
(CH C(OH) ),
64.92
(CH N ),
27.13
CH C(OH) ). HR LSIMS (thioglycerol) for the mix-
3
2
2
(
3
2
+
ture of 16a and 16b, calcd m/z for C H N O Br (MÀ
H-3pyr), 5.34 (s, 4H, CH N ), 3.24 (s, 12H, Me), 2.63 (t,
2
2
9
28
2
2
+
Br) 515.1334. Found m/z: 515.1335. Anal. for
C H N O Br 1.3H O (mixture of 16a and 16b): calcd
J=7.6 Hz, 4H, CH2Ph), 1.89 (q, J=7.6 Hz, 2H, C–
1
3
.
CH –C). C NMR (75.78 MHz): d 157.97 (C-4pyr),
2
9
28
2
2
2
2
2
C, 56.20; H, 4.98; N, 4.52. Found: C, 56.18; H, 5.06; N,
4
144.80 (C-1 ), 143.06 (C-2 ), 133.65 (C-4 ), 130.46
(C-3 ), 129.52 (C-2 ), 109.11 (C-3pyr), 61.54
Ph Ph
Ph
pyr
Ph
.56.
+
(
C). HR LSIMS (thioglycerol+Na ), calcd m/z for
CH N ), 40.39 (Me), 35.97 (CH ), 34.15 (C–CH2–
2 2Ph
0
nopyridinium) dibromide (17). Yield: 41%. Mp 255–
+
1
,1 -[Methylenebis(benzene-1,4-diylmethylene)]bis(4-cya-
+
C H N Br (MÀBr) 545.2280. Found m/z: 545.2281.
3
1
38
4
ꢁ
1
.
Anal. for C H N Br H O: calcd C, 57.77; H, 6.26; N,
31 38 4 2 2
2
57 C. H NMR (400.13 MHz): d 9.33 (d, J=6.6 Hz,
H, H-2pyr), 8.51 (d, J=6.6 Hz, 4H, H-3pyr), 7.51 (d,
J=8.1 Hz, 4H, Ph), 7.34 (d, J=8.1 Hz, 4H, Ph), 5.94 (s,
4
8.69. Found: C, 57.48; H, 6.41; N, 8.76.
+
13
0
bis(4-aminopyridinium) dibromide (27). Yield: 47%. Mp
4
H, CH N ), 4.07 (s, 2H, CH2Ph).
2
C
NMR
1,1 - [Propane - 1,3 - diylbis(benzene - 1,4 - diylmethylene)]-
(
100.03 MHz, selected data): d 144.64 (C-2pyr), 132.47
+
ꢁ
1
(
HR LSIMS (thioglycerol), calcd m/z for C H N Br
(
C H N Br H O: calcd C, 55.88; H, 4.17; N, 9.65.
C-3pyr), 115.18 (CN), 66.37 (CH N ), 42.02 (CH2Ph).
207–209 C. H NMR (300.13 MHz): d 8.17 (d, J=7.6
Hz, 4H, pyrH-2), 7.31 (d, J=8.3 Hz, 4H, Ph), 7.24 (d,
J=8.3 Hz, 4H, Ph), 6.86 (d, J=7.6 Hz, 4H, H-3pyr),
2
2
7
22
4
+
MÀBr) 481.1028. Found m/z: 481.1029. Anal. for
+
.
5.31 (s, 4H, CH N ), 2.63 (t, J=7.6 Hz, 4H, CH2Ph),
2
2
7
22
4
2
2
1
3
Found: C, 56.12; H, 3.93; N, 9.28.
1.90 (q, J=7.6 Hz, 2H, C–CH –C).
2
C NMR
(
75.78 MHz): d 160.79 (C-4pyr), 144.80 (C-1 ), 144.00
Ph
0
droxyiminomethyl)pyridinium] dibromide (18). Yield:
1
,1 -[Methylenebis(benzene-1,4-diylmethylene)]bis[(4-hy-
(C-2pyr), 133.59 (C-4 ), 130.47 (C-3 ), 129.46 (C-2 ),
Ph Ph Ph
110.95 (C-3pyr), 61.81 (CH N ), 35.98 (CH2Ph), 34.08
2
(C–CH –C). HR LSIMS (thioglycerol), calcd m/z for
2
+
ꢁ
1
4
(
8%. Mp 175–177 C. H NMR (300.13 MHz): d 8.97
d, J=7.0 Hz, 4H, H-2pyr), 8.30 (s, 2H, CH), 8.20 (d,
J=7.0 Hz, 4H, H-3pyr), 7.45 (d, J=8.3 Hz, 4H, Ph),
+
C H N Br (MÀBr) 489.1654. Found m/z: 489.1654.
27
30
4
.
Anal. for C H N Br 1.3H O: calcd C, 54.61; H, 5.53;
N, 9.43. Found: C, 54.76; H, 5.65; N, 9.25.
27
30
4
2
2
+
7
.31 (d, J=8.3 Hz, 4H, Ph), 5.78 (s, 4H, CH N ), 4.02
2
1
3
(
s, 2H, CH2 Ph). C NMR (75.78 MHz): d 151.50 (C-
4pyr), 145.87 (C-2pyr or CH), 145.58 (CH or C-2pyr),
44.26 (C-1 ), 132.62 (C-4 ), 131.15 (C-3 ), 130.48
0
bis[(4-hydroxymethyl)pyridinium] dibromide (28). Yield:
1,1 - [Propane - 1,3 - diylbis(benzene - 1,4 - diylmethylene)]-
1
Ph
Ph
Ph
+
ꢁ
ꢁ
1
(
C-2 ), 125.64 (C-3pyr), 64.84 (CH N ), 42.00 (CH2Ph).
Ph
70%. 132–134 C, 78–80 C dec.
H
NMR
2
HR LSIMS (thioglycerol), calcd m/z for C H N O
2
(300.13 MHz): d 8.98 (d, J=6.8 Hz, 4H, H-2pyr), 8.06
(d, J=6.8 Hz, 4H, H-3pyr), 7.44 (d, J=8.1 Hz, 4H, Ph),
7.28 (d, J=8.1 Hz, 4H, Ph), 5.80 (s, 4H, CH N ), 4.91
2
2
7
27
4
+
(
for C H N Br H O: calcd C, 52.62; H, 4.58; N, 9.09.
MÀ2Br+H) 439.2134. Found m/z: 439.2134. Anal.
.
+
2
7
26
4
2
2
Found: C, 52.96; H, 4.56; N, 8.66.
(s, 4H, CH OH), 2.65 (t, J=7.7 Hz, 4H, CH2 Ph), 1.90
2
1
3
(q, J=7.7 Hz, 2H, C–CH –C). C NMR (75.78 MHz):
2
0
pyridinium) dibromide (23a). Yield: 57% (mixture of 23a
1
,1 -[Ethylenebis(benzene-1,4-diylmethylene)]bis(4-acetyl-
d 165.21(C-4pyr), 145.58 (C-1 ), 145.19 (C-2pyr), 132.34
Ph
(C-4 ), 130.73 (C-3 ), 130.24 (C-2 ), 126.07 (C-3pyr),
Ph
Ph
Ph
ꢁ
+
2 2
and 23b). Mp 222–224 C dec (mixture of 23a and 23b).
1
64.85 (CH N ), 62.88 (CH OH), 36.01 (CH2Ph), 34.04
.
(C–CH –C). Anal. for C H N O Br 1.4H O: calcd C,
2 29 32 2 2 2 2
H NMR (300.13 MHz): d 9.26 (d, J=6.9 Hz, 4H, H-
2pyr), 8.49 (d, J=6.9 Hz, 4H, H-3pyr), 7.45 (d, J=8.1
55.68; H, 5.61; N, 4.48. Found: C, 55.69; H, 5.68; N,
4.63.
Hz, 4H, Ph), 7.30 (d, J=8.1 Hz, 4H, Ph), 5.90 (s, 4H,
+
CH N ), 2.94 (s, 2H, CH Ph), 2.75 (s, 6H, CH CO).
3
2
2
1
3
0
bis(4-methylpyridinium) dibromide (29). Yield: 75%.
C NMR (75.479 MHz, selected data): d 195.56
1,1 - [Propane - 1,3 - diylbis(benzene - 1,4 - diylmethylene)]-
+
(
CH CO), 65.71 (CH N ), 29.11 (CH ).
3
2
3
1
Very hygroscopic to determine its mp. H NMR
(300.13 MHz): d 8.90 (d, J=6.6 Hz, 4H, H-2pyr), 7.94
(d, J=6.6 Hz, 4H, H-3pyr), 7.44 (d, J=8.1 Hz, 4H, Ph),
0
dihydroxyethyl)]pyridinium} dibromide (23b). H NMR
1
,1 -[Ethylenebis(benzene-1,4-diylmethylene)]bis{[4-(1,1-
1
+
(
(
300.13 MHz): d 9.03 (d, J=6.9 Hz, 4H, H-2pyr), 8.18
d, J=6.9 Hz, 4H, H-3pyr), 7.44 (d, J=8.3 Hz, 4H, Ph),
7.28 (d, J=8.1 Hz, 4H, Ph), 5.76 (s, 4H, CH N ), 2.67
2
(s, 6H, Me), 2.65 (t, J=7.6 Hz, 4H, CH2Ph), 1.90 (q,

J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
2227
1
3
J=7.6 Hz, 2H, C–CH –C). C NMR (75.78 MHz): d
1
8.18 (m, 4H, H-3quin and H-6quin or H-7quin), 8.02 (t,
2H, H-7quin or H-6quin), 7.29 (d, J=8.2 Hz, 4H, Ph),
7.23 (d, J=8.2 Hz, 4H, Ph), 6.31 (s, 4H, CH N ), 2.60
2
(t, J=7.7 Hz, 4H, CH2 Ph), 1.90 (q, J=7.7 Hz, 2H, C–
.
CH –C). Anal. for C H N Br 0.6H O: calcd C,
2 35 32 2 2 2
2
61.69 (C-4pyr), 145.78 (C-1 ), 145.19 (C-2pyr), 132.34
Ph
+
(
6
C-4 ), 130.68 (C-2 ), 130.20–130.08 (C-3 or C-3Ph),
pyr
Ph
Ph
+
4.64 (CH N ), 35.97 (CH ), 34.00 (C–CH –C), 22.03
2
2Ph
2
.
Me). Anal. for C H N Br 1.5H O: calcd C, 58.50; H,
(
2
9
32
2
2
2
5
.93; N, 4.70. Found: C, 58.28; H, 5.92; N, 4.84.
64.55; H, 5.14; N, 4.30. Found: C, 64.70; H, 5.42; N,
4.45.
0
bis(4-acetylpyridinium) dibromide (30a). Yield: 75%
1
,1 -[Propane - 1,3 - diylbis(benzene -1,4 -diylmethylene)]-
0
llylamino)pyridinium] dibromide (37). Yield: 47%. Mp
1,1 -[Methylenebis(benzene-1,4-diylmethylene)]bis[(4-dia-
ꢁ
(
mixture of 30a and 30b). Mp 232–233 C dec (mixture
1
ꢁ
1
of 30a and 30b). H NMR (400.13 MHz): d 9.25 (d,
J=6.6 Hz, 4H, H-2pyr), 8.48 (d, J=6.6 Hz, 4H, H-3pyr),
112–114 C. H NMR (400.13 MHz): d 8.25 (d, J2,3=7.9
Hz, 4H, H-2pyr), 7.32 (d, J2,3=8.2 Hz, 4H, H-2benz),
7.26 (d, J2,3=8.2 Hz, 4H, H-3benz), 7.01 (d, J2,3=7.9 Hz,
7
.48 (d, J=8.1 Hz, 4H, Ph), 7.30 (d, J=8.1 Hz, 4H, Ph),
+
5
J=7.5 Hz, CH2 Ph), 1.91 (q, J=7.5 Hz, C–CH –C).
.91 (s, 4H, CH N ), 2.74 (s, 6H, CH CO), 2.66 (t, 4H,
1
4H, H-3pyr), 5.89 (ddt, J2,3
Hz, 4H, H-2allyl), 5.35 (s, 4H, CH N ), 5.27 (ddt,
0
=17.2, J2,3=10.2, J1,2=4.8
2
3
3
+
C
2
2
NMR (100.03 MHz, selected data): d 195.55 (CH CO),
J2,3=10.2, J3,3
(ddt, J2,3 =17.2, J3,3
.23 (dt, J1,2=4.8, J1,3=1.5, J1,3
0
=1.3, J1,3=1.5 Hz, 4H, H-3allyl), 5.21
3
+
0
6
5.79 (CH N ), 29.08 (CH ).
0
0
=1.3, J1,3
0
=1.5 Hz, 4H, H-3 allyl),
=1.5 Hz, 4H, H-1allyl),
2
3
4
0
0
bis[(4-(1,1-dihydroxyethyl)]pyridinium} dibromide (30b).
13
1
,1 -[Propane-1,3-diylbis(benzene-1,4-diylmethylene)]{-
3.99 (s, 2H, CH2Ph). C NMR (100.03 MHz): d 157.89
(C-4pyr), 143.66 (C-1 ), 143.49 (C-2pyr), 133.85 (C-4Ph),
131.47 (C-2allyl), 130.91 (C-3 ), 129.74 (C-2 ), 118.38
Ph
1
H NMR (400.13 MHz): d 9.04 (d, J=6.7 Hz, 4H, H-
2pyr), 8.18 (d, J=6.7 Hz, 4H, H-3pyr), 7.47 (d, J=8.2
Ph
Ph
+
(C-3allyl), 109.92 (C-3pyr), 61.70 (CH N ), 54.23 (C-1allyl),
2
+
Hz, 4H, Ph), 7.31 (d, J=8.2 Hz, 4H, Ph), 5.83 (s, 4H,
+
41.98 (CH2Ph). HR LSIMS (thioglycerol+Na ), calcd
+
CH N ), 1.62 (s, 6H, CH C(OH) ), 2.66 (t, 4H, J=7.7
2
m/z for C H N Br (MÀBr) 621.2593. Found m/z:
2
3
37 42
4
1
3
.
621.2593. Anal. for C H N Br 3.2H O: calcd C, 58.46;
37 42 4 2 2
Hz, CH2 Ph), 1.90 (q, J=7.6 Hz, C–CH –C). C NMR
2
(
100.03 MHz, selected data): d 98.14 (CH C(OH) ),
H, 6.42; N, 7.37. Found: C, 58.30; H, 6.20; N, 7.51.
3
2
+
2 3 2
6
C H N O Br 3.3H O (mixture of 30a and 30b): calcd
C, 54.45; H, 5.69; N, 4.10. Found: C, 54.28; H, 5.34; N,
4
5.09 (CH N ), 27.08 (CH C(OH) ). Anal. for
0 0
1,1 -[Biphenyl-3,3 -diylbis(methylene)]bispiridinium (38).
.
3
1
32
2
2
2
2
ꢁ
1
Yield: 57.3%. Mp 74–75 C. H NMR (400.13 MHz): d
9.25 (d, J=6.6 Hz, 4H, H-2pyr), 8.62 (dt, J=6.6 and 1.2
Hz, 2H, H-4pyr), 8.15 (m, 4H, H-3pyr), 8.06 (s, 2H, Ph),
7.78 (m, 2H, Ph), 7.57 (m, 4H, Ph), 5.98 (s, 4H,
.05.
0
bis(4-cyanopyridinium) dibromide (31). Yield: 29%. Mp
1
,1 -[Propane - 1,3 - diylbis(benzene -1,4 -diylmethylene)]-
+
13
CH N ). C NMR (100.03 MHz): d 147.34 (C-2pyr),
2
ꢁ
Hz, 4H, H-2pyr), 8.51 (d, J=6.5 Hz, 4H, H-3pyr), 7.50
1
2
40–241 C. H NMR (400.13 MHz): d 9.33 (d, J=6.5
146.08 (C-4pyr), 142.57 (C-1 ), 135.48 (C-3 ), 131.41
Ph
Ph
(C-5 ), 129.80 (C-3 ), 129.64, 129.62, 129.35 (C-
Ph
pyr
+
.
4,6,2 ), 65.53 (CH N ). Anal. for C H N Br 2H O:
Ph 2 24 22 2 2 2
(d, J=8.1 Hz, 4H, Ph), 7.32 (d, J=8.1 Hz, 4H, Ph), 5.93
+
(
(
s, 4H, CH N ), 2.67 (t, J=7.7 Hz, 4H, CH2Ph), 1.91
2
calcd C, 53.95; H, 4.91; N, 5.24. Found: C, 54.00; H,
4.55; N, 5.12.
1
3
q, J=7.7 Hz, 2H, C–CH –C). C NMR (100.03 MHz):
2
d 147.34 (C-2pyr), 146.10 (C-1 ), 132.47 (C-3pyr), 131.18
Ph
0
isoquinolinium dibromide (40). Yield: 90%. Mp 80–
82 C. H NMR (400.13 MHz): d 10.11 (s, 2H, H-1isoq),
8.67 (dd, J=6.8 and 1.2 Hz, 2H, H-3isoq), 8.51 (d,
J=8.3 Hz, 2H, H-5isoq or H-8isoq), 8.28 (d, J=8.2 Hz,
(
C-4 ), 130.88 (C-3 ), 130.80 (C-2 ), 130.12 (C-4pyr),
+
1,1 -[Propane-1,3-diylbis(benzene-1,4-diylmethylene)]bis-
Ph
Ph
Ph
1
CH –C). Anal. for C H N Br 0.3H O: calcd C,
15.22 (CN), 66.54 (CH N ), 36,02 (CH2Ph), 33.94 (C–
.
2
ꢁ
1
2
29 26
4
2
2
5
9
8.46; H, 4.50; N, 9.40. Found: C, 58.41; H, 4.40; N,
.15.
2H, H-8isoq or H-5isoq), 8.23 (ddd, J=8.2, 7.0 and 0.8
Hz, 2H, H-6isoq or H-7isoq), 8.05 (ddd, J=8.2, 7.0 and
0
0
trans-1,1 -[Stilbene-4,4 -diylbis(methylene)]bis(4-amino-
pyridinium) dibromide (33). Yield: 43%. Mp 329–
0.9 Hz, 2H, H-7isoq or H-6isoq), 7.50 (d, J=8.0 Hz, 4H,
+
Ph), 7.27 (d, J=8.0 Hz, 4H, Ph), 6.10 (s, 4H, CH N ),
ꢁ
1
3
4
4
5
31 C. H NMR (300.13 MHz): d 8.17 (d, J=7.6 Hz,
H, H-2pyr), 7.63 (d, J=8.4, 4H, Ph), 7.37 (d, J=8.4,
H, Ph), 7.24 (s, 2H, CH), 6.86 (d, J=7.6, 4H, H-3pyr),
.33 (s, 4H, CH N ). C NMR (75.78 MHz): d 160.90
C-4pyr), 144.06 (C-2pyr), 139.55 (C-1 ), 135.48 (C-4 ),
2
2.63 (t, J=7.7 Hz, 4H, CH2Ph), 1.88 (q, J=7.7 Hz, 2H,
.
C–CH –C). Anal. for C H N Br 2H O: calcd C,
2
35 32
2
2
2
+
13
62.14; H, 5.36; N, 4.14. Found: C, 62.04; H, 5.34; N,
4.17.
2
(
Ph Ph
1
3pyr), 61.73 (CH N ). Anal. for C H N Br 1.5H O:
29.91 (CH), 129.74 (C-3 ), 128.55 (C-2 ), 110.99 (C-
Ph Ph
+
0
bis[(4-dimethylamino)pyridinium] dibromide (41). Yield:
89%. Mp 100–103 C. H NMR (300.13 MHz): d 8.32
(d, J=7.9 Hz, 4H, H-2pyr), 7.39 (d, J=8.2 Hz, 4H, Ph),
.
1,1 - [Butane - 1,4 - diylbis(benzene - 1,4 - diylmethylene)]-
2
29 32
2
2
2
calcd C, 58.50; H, 5.93; N, 4.70. Found: C, 58.28; H,
5
ꢁ
1
.92; N, 4.84. Anal. for C H N Br : calcd C, 56.33; H,
2
6
26
4
2
4
.73; N, 10.11. Found: C, 56.05; H, 4.48; N, 10.15.
7
.31 (d, J=8.2 Hz, 4H, Ph), 7.09 (d, J=7.9 Hz, 4H, H-
+
0
1
quinolinium dibromide (34). Yield: 36%. Mp 158–160 C
,1 -[Propane-1,3-diylbis(benzene-1,4-diylmethylene)]bis-
ꢁ
3pyr), 5.42 (s, 4H, CH N ), 3.33 (s, 12H, NMe ), 2.72 (t,
2 2
4H, CH2
(75.78 MHz): d 157.98 (C-4pyr), 145.13 (C-1 ), 143.05
13
), 1.70 (q, 4H, C–CH –C). C NMR
Ph 2
1
dec. H NMR (400.13 MHz):d 9.52 (d, J=5.9 Hz, 2H,
H-2quin), 9.28 (d, J=8.3 Hz, 2H, H-4quin), 8.52 (d, 2H,
H-5quin or H-8quin), 8.45 (d, 2H, H-8quin or H-5quin),
Ph
(C-2pyr), 133.51 (C-4 ), 130.42 (C-3 ), 129.44 (C-2 ),
+
Ph
Ph
Ph
109.11 (C-3pyr), 61.57 (CH N ), 40.39 (NMe ), 36.28
2 2

2228
J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
(
CH2Ph), 32.05 (C–CH –C). HR LSIMS (thioglycer-
2
C, 58.46; H, 5.38; N, 5.05. Found: C, 58.54; H, 5.47; N,
5.11.
+
+
ol+Na ), calcd m/z for C H N Br (MÀBr) 559.2436.
3
2
40
4
.
Found m/z: 559.2436. Anal. for C H N Br 1.2H O:
32
40
4
2
2
0
0
calcd C, 58.05; H, 6.46; N, 8.46. Found: C, 57.86; H, 6.35;
N, 8.26.
trans-1,1 -[Stilbene-4,4 -diylbis(methylene)]bis[(4-dime-
thylamino)pyridinium] dibromide (46). Yield: 81%. Mp
>
ꢁ
ꢁ
1
315 C, 190–192 C dec. H NMR (400.13 MHz): d
0
0
cis-1,1 -[Stilbene-4,4 -diylbis(methylene)]bis[(4-dimethyl-
amino)pyridinium) dibromide (42). Yield: 61%. Mp 133–
1
4
3pyr), 6.66 (s, 2H, CH), 5.34 (s, 4H, CH N ), 3.25 (s,
1
8.25 (d, J=6.5 Hz, 4H, H-2pyr), 7.63 (d, J=8.3 Hz, 4H,
Ph), 7.39 (d, J=8.3 Hz, 4H, Ph), 7.23 (s, 2H, CH), 7,00
ꢁ
H, H-2pyr), 7.26 (s, 8H, Ph), 7.01 (d, J=7.9 Hz, 4H, H-
1
+
35 C. H NMR (300.13 MHz): d 8.23 (d, J=7.9 Hz,
(d, J=6.5 Hz, 4H, H-3 ), 5.36 (s, 4H, CH N ), 3.24
pyr
2
1
3
(s, 12H, CH3). C NMR (100.03 MHz): d 158.01 (C-
+
4pyr), 143.09 (C-2pyr), 139.52 (C-1 ), 135.49 (C-4 ),
129.91 (CH), 129.83 (C-3 ), 128.53 (C-2 ), 109.16 (C-
Ph Ph
2
Ph
Ph
1
3
2H, CH3). C NMR (75.78 MHz): d 158.00 (C-4pyr),
+
1
43.11 (C-2pyr), 139.38 (C-1 ), 135.16 (C-4 ), 131.32
3pyr), 61.47 (CH N ), 40.38 (Me). HR LSIMS (thio-
2
Ph
Ph
+
+
(CH), 130.79 (C-3 ), 129.40 (C-2 ), 109.18 (C-3pyr),
Ph
glycerol+Na ), calcd m/z for C H N Br (MÀ Br)
Ph
30 34
4
+
6
1.36 (CH N ), 40.42 (Me). HR LSIMS (thioglycerol),
2
529.1934.
Found
m/z:
529.1934.
Anal.
for
+
.
C H N Br 1.3H O 0.5CH OH: calcd C, 56.37; H,
30 34 4 2 3
.
calcd m/z for C H N Br (MÀBr) 529.1967. Found
3
0
34
4
2
.
m/z: 529.1968. Anal. for C H N Br H O: calcd C,
2
5.99; N, 8.62. Found: C, 56.28; H, 5.85; N, 8.27.
3
0
34
4
2
5
8
7.34; H, 5.77; N, 8.92. Found: C, 57.62; H, 5.92; N,
.81.
0
0
cis-1,1 -[Stilbene-4,4 -diylbis(methylene)]bis(4-aminopyri-
ꢁ
1
dinium) dibromide (47). Yield: 40%. Mp 273–275 C. H
NMR (300.13 MHz): d 8.17 (d, J=7.5 Hz, 4H, H-2pyr),
7.25 (s, 8H, Ph), 6.87 (d, J=7.5 Hz, 4H, H-3pyr), 6.66 (s,
2H, CH), 5.31 (s, 4H, CH N ). C NMR (75.78 MHz):
2
d 160.84 (C-4pyr), 144.06 (C-2pyr), 139.37 (C-1 ), 135.13
0 0
,1 -[Biphenyl-4,4 -diylbis(methylene)]bispyridinium dibro-
1
ꢁ
1
mide (43). Yield: 98%. Mp 275–277 C. H NMR
300.13 MHz): d 9.15 (dd, J=6.6 and 1.2 Hz, 4H, H-
2pyr), 8.64 (dt, J=9.1 and 1.3 Hz 2H, H-4pyr), 8.16 (m,
H, H-3pyr), 7.75 (d, J=8.5 Hz, 4H, Ph), 7.64 (d, J=8.5
+
13
(
Ph
4
(C-4 ), 131.34 (CH), 130.80 (C-3 ), 129.35 (C-2 ),
Ph
Ph
Ph
+
13
+
.
111.01 (C-3pyr), 61.63 (CH N . Anal. for C H N Br
2
Hz, 4H, Ph), 5.94 (s, 4H, CH N ).
2
C NMR
75,78 MHz): d 147.39 (C-2pyr), 146.05 (C-4pyr), 142.79
2
26 26
4
(
0.5H O: calcd C, 55.81; H, 4.95; N, 9.82. Found: C,
2
(
1
C-1 ), 134.24 (C-4 ), 130.91 (C-3 ), 129.78 (C-3pyr),
Ph
55.67; H, 4.57; N, 9.44.
Ph
Ph
+
29.26 (C-2 ), 65.32 (CH N ). HR LSIMS (thiogly-
Ph 2
+
0
cerol), calcd m/z for C H N (MÀ 2Br+H)
1,1 -[Methylenebis(benzene-1,4-diylmethylene)]bispyridi-
2
4
23
2
ꢁ
1
3
39.1861.
Found
m/z:
339.1860.
Anal.
C H N Br 0.8H O: calcd C, 56.23; H, 4.64; N, 5.46.
for
nium dibromide (49). Yield: 82%. Mp 243–245 C. H
NMR (300.13 MHz): d 9.09 (dd, J=6.6 and 1.2 Hz, 4H,
H-2pyr), 8.61 (dt, J=9.1 and 1.3 Hz, 2H, H-4pyr), 8.12
.
2
4
22
2
2
2
Found: C, 56.11; H, 4.36; N, 5.24.
(
m, 4H, H-3pyr), 7.47 (d, J=8.2 Hz, 4H, Ph), 7.32 (d,
+
0
bis[(4-pyrrolidino)pyridinium) dibromide (44). Yield:
1
,1 -[Propane - 1,3 - diylbis(benzene -1,4 -diylmethylene)]-
J=8.2 Hz, 4H, Ph), 5.84 (s, 4H, CH N ), 4.02 (s, 2H,
2
CH2 Ph). C NMR (75.78 MHz): d 147.26 (C-2pyr),
145.90 (C-4pyr), 144.31 (C-1 ), 132.59 (C-4 ), 131.15
(C-3 ), 130.50 (C-2 ), 129.71 (C-3pyr), 65.41
Ph Ph
(CH N ), 41,99 (CH ). HR LSIMS (thioglycerol),
2 2Ph
1
3
ꢁ
ꢁ
1
4
9%. Mp 182–186 C, dec at 158 C. H NMR
Ph
Ph
(
(
(
(
400.13 MHz): d 8.21 (d, J=7.5 Hz, 4H, H-2pyr), 7.31
+
d, J=8.0 Hz, 4H, Ph), 7.24 (d, J=8.0 Hz, 4H, Ph), 6.85
+
+
d, J=7.5 Hz, 4H, H-3pyr), 5.32 (s, 4H, CH N ), 3.54
2
calcd m/z for C H N (MÀ2Br+H)
353.2018.
25 25
2
.
Found m/z: 353.2018. Anal. for C H N Br 1.3H O:
25 24 2 2 2
t, J=6.7 Hz, 8H, H-2pyrrolid), 2.63 (t, J=7.6 Hz, 4H,
CH2Ph), 2.11 (q, J=6.7 Hz, 8H, H-3pyrrolid), 1.89 (q,
1
calcd C, 56.05; H, 5.00; N, 5.23. Found: C, 56.26; H,
4.87; N, 5.24.
3
J=7.6 Hz, 2H, C–CH –C). C NMR (100.03 MHz): d
2
1
55.14 (C-4pyr), 144.77 (C-1 ), 142.98 (C-2pyr), 133.77
Ph
0
nium dibromide (50). Yield: 34%. Mp 180–182 C, 154–
156 C dec. H NMR (300.13MHz): d 9.54 (d, 2H,
H-2quin), 9.30 (d, 2H, H-4quin), 8.51 (d, 2H, H-5quin or H-
8quin), 8.45 (d, 2H, H-8quin or H-5quin), 8.19 (m, 4H, H-3quin
and H-6quin or H-7quin), 8.02 (t, 2H, H-7quin or H-6quin),
(
C-4 ), 130.45 (C-3 ), 129.46 (C-2 ), 109.68 (C-3pyr),
Ph
1,1 -[Ethylenebis(benzene - 1,4- diylmethylene)]bisquinoli-
ꢁ
Ph
Ph
+
6
1.57 (CH N ), 49.72 (C-2pyrrolid), 35.99 (CH2Ph), 34.20
2
ꢁ
1
(
C–CH –C), 26.13 (C-3
). HR LSIMS (thioglycer-
pyrrolid
2
+
+
ol+Na ), calcd m/z for C H N Br (MÀBr)
3
5
42
4
5
97.2593.
Found
m/z:
597.2593.
Anal.
for
C H N Br 3H O: calcd C, 57.38; H, 6.60; N, 7.65.
.
3
5
42
4
2
2
Found: C, 57.19; H, 6.47; N, 7.75.
7.26 (d, J=8.4 Hz, 4H, Ph), 7.21 (d, J=8.4 Hz, 4H, Ph),
+
6
.32 (s, 4H, CH N ), 2.87 (s, 4H, CH2 Ph). HR LSIMS
2
0
pyridinium dibromide (45). Yield: 76%. Mp 62–64 C.
+
1
,1 -[Propane-1,3-diylbis(benzene-1,4-diylmethylene)]bis-
ꢁ
(thioglycerol), calcd m/z for C H N (M- 2Br+H)
34 31 2
467.2487.
Found
.
m/z:
467.2489.
Anal.
for
C H N Br 1.8H O: calcd C, 61.98; H, 5.14; N, 4.25.
1
H NMR (300.13 MHz): d 9.10 (dd, J=6.6 and 1.2 Hz,
3
4
30
2
2
2
4
8
H, H-2pyr), 8.61 (dt, J=9.1 and 1.3 Hz 2H, H-4pyr),
,13 (m, 4H, H-3pyr), 7.47 (d, J=8.2 Hz, 4H, Ph), 7.29
Found: C, 61.18; H, 5.17; N, 4.07.
+
0 0
1,1 -[Biphenyl-4,4 -diylbis(methylene)]bisquinolinium di-
bromide (51). Yield: 71%. Mp 247–249 C, 178–181 C
(d, J=8.2 Hz, 4H, Ph), 5.85 (s, 4H, CH N ), 2.65 (t,
J=7.6 Hz, 4H, CH Ph), 1.92 (q, J=7.6 Hz, 2H, C-CH2-
C). C NMR (75.78 MHz): d 147.21 (C-2pyr), 145.87 (C-
2
ꢁ
ꢁ
2
1
3
1
dec. H NMR (300.13 MHz): d 9.61 (d, 2H, H-2quin),
4pyr), 145.65 (C-1 ), 132.09 (C-4 ), 130.73 (C-3 ), 130.33
+
9.31 (d, 2H, H-4quin), 8.53 (d, 2H, H-5quin or H-8quin),
8.46 (d, 2H, H-8quin or H-5quin), 8.21 (m, 4H, H-3quin
and H-6quin or H-7quin), 8.02 (t, 2H, H-7quin or H-6quin),
Ph
Ph
Ph
(
C-2 ), 129.68 (C-3 ), 65.52 (CH N ), 35.98 (CH2Ph),
Ph pyr 2
.
33.97 (C–CH –C). Anal. for C H N Br 0.8H O: calcd
2 27 28 2 2 2

J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
2229
0
dibromide (56). Yield: 80%. Mp 247–249 C, 179–
181 C dec. H NMR (300.13 MHz):d 10.17 (s, 2H, H-
1isoquin), 8.73 (d, J=6.8 Hz, 2H, H-3isoquin), 8.54 (d,
0
7
.66 (d, J=8.4 Hz, 4H, Ph), 7.46 (d, J=8.4 Hz, 4H, Ph),
+
1,1 - [Biphenyl - 4,4 - diylbis(methylene)]bisisoquinolinium
ꢁ
.
.41 (s, 4H, CH N ). Anal. for C H N Br 1.9H O:
6
calcd C, 60.76; H, 4.75; N, 4.43. Found: C, 61.00; H,
4
2
32 26
2
2
2
ꢁ
1
.77; N, 4.12.
2
H, H-5isoquin or H-8isoquin), 8.50 (d, J=6.8 Hz, 2H,
0
rolidino)pyridinium] dibromide (52). Yield: 72%. Mp
1
,1 -[Methylenebis(benzene-1,4-diylmethylene)]bis[(4-pyr-
H-4isoquin), 8.31 (d, 2H, H-8isoquin or H-5isoquin), 8.25 (t,
2H, H-6isoquin or H-7isoquin), 8.07 (t, 2H, H-7isoquin or
H-6isoquin), 7.73 (d, J=8.5 Hz, 4H, Ph), 7.69 (d, J=8.5
ꢁ
Hz, 4H, H-2pyr), 7.31 (d, J=8.1 Hz, 4H, Ph), 7.25 (d,
1
1
39–141 C. H NMR (400.13 MHz): d 8.19 (d, J=7.6
+
Hz, 4H, Ph), 6.05 (s, 4H, CH N ). HR LSIMS (thio-
2
+
J=8.1 Hz, 4H, Ph), 6.84 (d, J=7.6 Hz, 4H, H-3pyr),
+
glycerol), calcd m/z for C H N Br (MÀBr)
32
26
2
5.32 (s, 4H, CH N ), 3.97 (s, 2H, CH2Ph), 3.54 (t,
J=6.8 Hz, 8H, H-2pyrrolid), 2.11 (q, J=6.8 Hz, 8H, H-
517.1279.
Found
m/z:
517.1278.
Anal.
for
2
.
C H N Br 0.5H O: calcd C, 63.28; H, 4.48; N, 4.61.
32 26 2 2 2
1
3
3pyrrolidin). C NMR (100.03 MHz): d 155.13 (C-4pyr),
43.58 (C-1 ), 142.97 (C-2 ), 134.22 (C-4 ), 130.85
Found: C, 63.36; H, 4.75; N, 4.55.
1
Ph
pyr
Ph
0
methylanilino)pyridinium] dibromide (57). Yield: 70%.
(
(
C-3 ), 129.62 (C-2 ), 109.71 (C-3pyr), 61.47
Ph
1,1 -[Methylenebis(benzene-1,4-diylmethylene)]bis[(4-N-
Ph
+
CH N ), 49.74 (C-2pyrrolid), 41.99 (CH ), 26.14 (C-
2
2Ph
+
ꢁ
H-2pyr, T =1.7 s), 7.58 (t, J2,3=6.7 Hz, 4H, H-3anil),
1
3pyrrolid). HR LSIMS (thioglycerol+Na ), calcd m/z
+
Mp 116–117 C. H NMR (300.13 MHz): d 8.28 (s, 4H,
for C H N Br (MÀBr)
569.2280. Found m/z:
3
3
38
4
1
.
69.2279. Anal. for C H N Br 3H O: calcd C, 56.26;
5
H, 6.30; N, 7.95. Found: C, 56.22; H, 6.35; N, 7.91.
7.48 (t, J3,4=7.4 Hz, 2H, H-4anil), 7.34 (d, J2,3=7.7 Hz,
4H, H-2anil), Hanil T =2.5 s, and 2.7 s, 7.33 (d, J2,3=8.1
3
3
38
4
2
2
1
Hz, 4H, H-2benz), 7.25 (d, J2,3=8.1 Hz, 4H, H-3benz),
+
6.90 (s, 4H, H-3pyr, T =1.6 s), 5.37 (s, 4H, CH N ),
3.97 (s, 4H, CH Ph), 3.52 (s, 6H, CH N). C NMR
2 3
0
1
linium dibromide (53). Yield: 91%. Mp 258–260 C. H
,1 -[Ethylenebis(benzene-1,4-diylmethylene)]bisisoquino-
ꢁ
1
2
1
13
NMR (300.13 MHz): d 10.09 (s, 2H, H-1isoquin), 8.66
(75.57 MHz): d 158.37 (C-4pyr), 144.79 (C-1anil), 144.22
(
dd, J=6,9 and 1.5 Hz, 2H, H-3isoquin), 8.48 (d, J=6.9
(C-1 ), 143.65 (C-2 ), 133.90 (C-4 ), 132.00 (C-3 ),
Ph
pyr
Ph
Ph
Hz, 2H, H-4isoquin), 8.52 (dd, J=8.3 and 0.9 Hz, 2H, H-
5isoquin or H-8isoquin), 8.30 (d, J=8.0 Hz, 2H, H-8isoquin
or H-5isoquin), 8.25 (dt, J=6.6 and 1.2 Hz, 2H, H-6isoquin
or H-7isoquin), 8.07 (dt, J=6.6 and 1.2 Hz, 2H, H-7isoquin
130.87, 130.08, 129.78 (C-3anil, C-2anil, C-4anilino or C-
+
2Ph), 127.51 (C-4anil), 110.25 (C-3pyr), 61.76 (CH N ),
2
41.97 (Ph–CH –Ph), 41.47 (CH N). HR LSIMS (thio-
3
2
+
+
glycerol+Na ), calcd m/z for C H N Br (MÀBr)
39
38
4
or H-6isoquin), 7.48 (d, J=8.1 Hz, 4H, Ph), 7.29 (d,
+
641.2280.
Found
m/z:
641.2279.
Anal.
for
.
C H N Br 0.8H O: calcd C, 63.52; H, 5.77; N, 7.50.
39 38 4 2 2
J=8.1 Hz, 4H, Ph), 5.94 (s, 4H, CH N ), 2.93 (s, 4H,
2
CH2
C H N Br (MÀBr) 545.1592. Found m/z: 545.1593.
). HR LSIMS (thioglycerol), calcd m/z for
+
Found: C, 63.56; H, 5.41; N, 7.60.
Ph
3
4
30
2
0
thylanilino)pyridinium] dibromide (58). Yield: 22%. Mp
.
Anal. for C H N Br H O: calcd C, 63.37; H, 5.00; N,
2
1,1 -[Ethylenebis(benzene-1,4-diylmethylene)]bis[4-(N-me-
3
4
30
2
2
4
.35. Found: C, 63.29; H, 4.84; N, 4.29.
ꢁ
1
1
15–116 C. H NMR (400.13 MHz, room tempera-
0
nolinium dibromide (54). Yield: 83%. Mp 277–279 C.
1
,1 -[Methylenebis(benzene-1,4-diylmethylene)]bisisoqui-
ꢁ
ture): d 8.26 (s, 4H, H-2pyr, T =1.6 s), 7.58 (t, J2,3=7.6
1
Hz, 4H, H-3anil), 7.49 (t, J3,4=7.4 Hz, 2H, H-4anil), 7.35
(d, J2,3=7.7 Hz, 4H, H-2anil), Hanil T =2.8 s, 2.9 s, and
1
H NMR (300.13 MHz): d 10.09 (s, 2H, H-1isoquin), 8.65
dd, J=6,8 and 1.5 Hz, 2H, H-3isoquin), 8.50 (dd, J=8.3
1
(
3.0 s, 7.31 (d, J2,3=8.2 Hz, 4H, H-2benz), 7.24 (d,
J2,3=8.2 Hz, 4H, H-3 ), 6.89 (s, 4H, H-3pyr, T =1.6
and 1.0 Hz, 2H, H-5isoquin or H-8isoquin), 8.44 (d, J=6.8
Hz, 2H, H-4isoquin), 8.27 (d, J=7.7 Hz, 2H, H-8isoquin or
H-5isoquin), 8.22 (ddd, J=7.6, 6.7 and 1.0 Hz, 2H, H-
6isoquin or H-7isoquin), 8.04 (ddd, J=8.3, 6.7 and 1.5 Hz,
benz
+
1
s), 5.36 (s, 4H, CH N ), 3.53 (s, 6H, CH N), 2.91 (s,
2
3
1
3
4H, CH2 Ph). C NMR (100.13 MHz, room tempera-
ture): d 158.45 (C-4pyr), 144.80 (C-1anil), 144.21 (C-1Ph),
143.63 (C-2pyr), 133.52 (C-4 ), 132.01 (C-3 ), 130.57,
2
Ph), 7.30 (d, J=8.2 Hz, 4H, Ph), 5.94 (s, 4H, CH N ),
H, H-7isoquin or H-6isoquin), 7.50 (d, J=8.2 Hz, 4H,
+
Ph
Ph
130.11, 129.53 (C-3anil, C-2anil, C-2 ), 127.48 (C-4anil),
Ph
2
+
3
.99 (s, 2H, CH2 Ph). HR LSIMS (thioglycerol), calcd
+
110.26 (C-3pyr), 61.92 (CH N ), 41.43 (CH N), 38.23
2
3
m/z for C H N Br (MÀBr) 531.1436. Found m/z:
(CH Ph).
2
3
3
28
2
.
31.1435. Anal. for C H N Br H O: calcd C, 62.87;
33 28 2 2 2
5
1
ꢁ
H, 4.80; N, 4.44. Found: C, 62.87; H, 4.66; N, 4.34.
H NMR (400.13 MHz, DMSO-d , 57 C): d 8.45 (d,
6
J2,3=7.5 Hz, 4H, H-2pyr), 7.59 (t, J2,3=7.7 Hz, 4H, H-
3anil), 7.48 (d, J3,4=7.4 Hz, 4H, H-4anil), 7.40 (d,
J2,3=7.4 Hz, 4H, H-2anil), 7.34 (d, J2,3=8.1 Hz, 4H, H-
0
nium dibromide (55). Yield: 30%. Mp 167–169 C. H
1
,1 -[Methylenebis(benzene-1,4-diylmethylene)]bisquinoli-
ꢁ
1
NMR (300.13 MHz): d 9.53 (d, 2H, H-2quin), 9.28 (d,
2benz), 7.29 (d, J2,3=8.1 Hz, 4H, H-3 ), 6.91 (d, 4H,
benz
+
+
2
2
H, H-4quin), 8.50 (d, 2H, H-5quin or H-8quin), 8.44 (d,
H, H-8quin or H-5quin), 8.18 (m, 4H, H-3quin and H-
H-3pyr), 5.36 (s, 4H, CH N ), 3.53 (s, 6H, CH N ),
2 3
1
3
2.91 (s, 4H, CH2 Ph). C NMR (100.13 MHz, DMSO-
d , 57 C): d 156.16 (C-4pyr), 143.18 (C-1anil), 142.59 (C-
ꢁ
2pyr), 142.08 (C-1 ), 132.92 (C-4 ), 130.53 (C-3anil),
6quin or H-7quin), 8.01 (t, 2H, H-7quin or H-6quin), 7.30
d, J=8.3 Hz, 4H, Ph), 7,24 (d, J=8.3, 4H, Ph), 6.31 (s,
6
(
4
Ph
Ph
+
H, CH N ), 3.95 (s, 2H, CH
2
). HR LSIMS (thio-
+
128.88 (C-3 ), 128.47 (C-4 ), 128.09 (C-2 ), 126.25
Ph anil Ph
2 Ph
+
glycerol), calcd m/z for C H N Br (MÀBr)
(C-2anil), 108.92 (C-3pyr), 59.41 (CH N ), 40.76
2
3
3
28
2
5
31.1436.
Found
m/z:
531.1436.
Anal.
C H N Br 2.3H O: calcd C, 60.62; H, 5.03; N, 4.28.
for
(CH N), 36.21 (CH2Ph). HR LSIMS (thioglycer-
3
.
+
+
ol+Na ), calcd m/z for C H N Br (MÀBr)
3
3
28
2
2
2
40 40
4
Found: C, 60.50; H, 4.77; N, 4.05.
655.2436.
Found
m/z:
655.2435.
Anal.
for

2230
J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
.
C H N Br 1.5H O: calcd C, 63.09; H, 5.95; N, 7.48.
Found: C, 62.92; H, 5.67; N, 7.34.
.
C H N Br 4H O: calcd C, 58.36; H, 6.78; N, 6.98.
39 46 4 2 2
Found: C, 58.27; H, 6.40; N, 6.93.
4
0
40
4
2
2
0
0
hydroazepino)pyridinium] dibromide (62). Yield: 58%.
1
,1 -[Propane - 1,3 - diylbis(benzene -1,4 -diylmethylene)]-
1,1 -[Methylenebis(benzene-1,4-diylmethylene)]bis[(4-per-
bis[(4-N-methylanilino)pyridinium)
dibromide
Yield: 30%. Mp 120–121 C. H NMR (300.13 MHz): d
.27 (s, 4H, H-2pyr, T =1.5 s), 7.59 (t, J2,3=7.5 Hz, 4H,
(59).
ꢁ
1
ꢁ
1
Mp 168–170 C. H NMR (300.13 MHz): d 8.19 (d,
J2,3=7.9 Hz, 4H, H-2pyr), 7.32 (d, J2,3=8.2 Hz, 4H, H-
2benz), 7.26 (d, J2,3=8.2 Hz, 4H, H-3benz), 7.04 (d,
8
1
H-3anil), 7.51 (t, J3,4=7.3 Hz, 2H, H-4anil), 7.32 (d,
J2,3=7.5 Hz, 4H, H-2anil), Hanil T =2.5 s, and 2.5 s,
+
J2,3=7.9 Hz, 4H, H-3pyr), 5.31 (s, 4H, CH N ), 3.99 (s,
2
1
7
4
.28 (d, J2,3=8.1 Hz, 4H, H-2benz), 7.21 (d, J2,3=8.0 Hz,
H, H-3benz), 6.88 (s, 4H, H-3pyr T =1.4 s), 5.38 (s, 4H,
2H, CH Ph), 3.71 (t, 8H, H-2
2
H-3perhydroazep), 1.59 (m, 8H, H-4perhydroazep). C NMR
(75.78 MHz): d 157.18 (C-4pyr), 143.61 (C-1 ), 143.33
), 1.84 (m, 8H,
perhydroazep
1
3
1
+
CH N ), 3.51 (s, 6H, CH N), 3.47 (t, J=7.1 Hz, 4H,
2
Ph–CH –C), 1.17 (q, J=7.1 Hz, 2H, C-CH -C).
2
3
Ph
1
3
C
(C-2pyr), 134.05 (C-4 ), 130.89 (C-3 ), 129.70 (C-2 ),
Ph
2
Ph
Ph
+
NMR (75.78 MHz): d 158.37 (C-4pyr), 144.78 (C-1anil),
44.21 (C-1 ), 143.76 (C-2 ), 133.60 (C-4 ), 132.00
109.03 (C-3pyr), 61.40 (CH N ), 51.32 (C-2perhydroazep),
2
1
41.98 (Ph–CH –Ph), 27.34 (C-3
2
), 24.93 (C-
perhydroazep
Ph
pyr
Ph
+
(C-3 ), 130.48, 130.08, 129.60 (C-3anil, C-2anil, C-2Ph or
C-2anil), 127.50 (C-2anil), 110.29 (C-3pyr), 61.89
(CH N ), 41.44 (CH N), 35.97 (Ph–CH –C), 15.46 (C–
CH –C). HR LSIMS (thioglycerol+Na ), calcd m/z
4perhydroazep). HR LSIMS (thioglycerol+Na ), calcd
+
Ph
m/z for C H N Br (MÀBr) 625.2906. Found m/z:
37
46
4
+
.
625.2904. Anal. for C H N Br 3.5H O: calcd C,
37 46 4 2 2
2
3
2
+
57.74; H, 6.94; N, 7.28. Found: C, 57.58; H, 6.81; N,
7.32.
2
+
for C H N Br (MÀBr)
669.2593. Found m/z:
4
1
42
4
.
69.2593. Anal. for C H N Br 4H O: calcd C, 60.15;
41 42 4 2 2
6
0
bis[(4-piperidino)pyridinium) dibromide (63). Yield:
H, 6.02; N, 6.99. Found: C, 59.86; H, 6.13; N, 6.81.
1,1 - [Propane - 1,3 - diylbis(benzene - 1,4 - diylmethylene)]-
0
bis[(4-perhydroazepino)pyridinium)
ꢁ
1
1
,1 -[Propane - 1,3 - diylbis(benzene -1,4 -diylmethylene)]-
dibromide
58%. Mp 86–88 C. H NMR (300.13 MHz): d 8.19 (d,
J2,3=7.9 Hz, 4H, H-2pyr), 7.31 (d, J2,3=8.2 Hz, 4H, H-
2benz), 7.25 (d, J2,3=8.2 Hz, 4H, H-3benz), 7.12 (d,
(60).
1
Yield: 61%. Very hygroscopic to determine its mp. H
NMR (400.13 MHz): d 8.20 (d, J2,3=7.9 Hz, 4H, H-
2pyr), 7.32 (d, J2,3=8.2 Hz, 4H, H-2benz), 7.25 (d,
+
J2,3=7.9 Hz, 4H, H-3pyr), 5.30 (s, 4H, CH N ), 3.70 (t,
2
8H, H-2perhydroazep), 2.64 (t, J=7.7 Hz, 4H, CH2Ph),
1.90 (q, J=7.7 Hz, 2H, C-CH -C), 1.75 (m, 12H, H-
J2,3=8.2 Hz, 4H, H-3benz), 7.04 (d, J2,3=7.9 Hz, 4H, H-
+
2
13
3pyr), 5.32 (s, 4H, CH N ), 3.72 (t, 8H, H-2perhydroazep),
2
3perhydroazep
and
H-4perhydroazep).
C
NMR
2
.64 (t, J=7.7 Hz, 4H, CH2 Ph), 1.90 (q, J=7.7 Hz, 2H,
C–CH –C), 1.84 (m, 8H, H-3perhydroazep), 1.58 (m, 8H,
(75.78 MHz): d 156.84 (C-4pyr), 144.82 (C-1 ), 143.49
Ph
(C-2pyr), 133.61 (C-4 ), 130.47 (C-3 ), 129.50 (C-2 ),
+
2
Ph
Ph
Ph
1
3
H-4perhydroazep). C NMR (100.13 MHz): d 157.22 (C-
109.26 (C-3pyr), 61.41 (CH N ), 49.08 (C-2perhydroazep),
2
4pyr), 144.86 (C-1 ), 143.33 (C-2pyr), 133.55 (C-4Ph),
1
35.99 (CH2Ph), 34.15 (C–CH –C), 26.66 (C-3
),
perhydroazep
24.93 (C-4perhydroazep). HR LSIMS (thioglycerol+Na ),
Ph
2
+
30.48 (C-3 ), 129.55 (C-2 ), 109.02 (C-3pyr), 61.55
Ph
Ph
+
+
(CH N ), 51.33 (C-2perhydroazep), 35.99 (CH2Ph), 34.05
2
calcd m/z for C H N Br (MÀ Br) 625.2906. Found
37 46
4
.
m/z: 625.2907. Anal. for C H N Br 3H O: calcd C,
37 46 4 2 2
(
HR LSIMS (thioglycerol+Na ), calcd m/z for
C-CH -C), 27.35 (C-3
), 24.93 (C-4perhydroazep).
perhydroazep
+
2
58.42; H, 6.89; N, 7.37. Found: C, 58.40; H, 6.62; N,
7.26.
+
C H N Br (MÀ Br) 597.2593. Found m/z: 597.2593.
3
5
42
4
+
HR LSIMS (thioglycerol+Na ), calcd m/z for
C H N Br (MÀBr) 653.3219. Found m/z: 653.3221.
+
3
9
50
4
Computational methods
.
Anal. for C H N Br 4H O: calcd C, 58.07; H, 7.25;
N, 6.94. Found: C, 57.80; H, 7.03; N, 7.18.
3
9
50
4
2
2
Statistical analysis was performed by partial least-
squares algorithm using the QSAR module of SYBYL
0
bis[(4-diallylamino)pyridinium] sibromide (61). Yield:
35
1
,1 -[Propane - 1,3 - diylbis(benzene -1,4 -diylmethylene)]-
software. The clog P values were calculated by using
the Ghose–Crippen modified atomic contribution sys-
1
26
6
7%. Very hygroscopic to determine its mp. H NMR
tem (ATOMIC5 option) contained in the PALLAS
2
7
(
(
400.13 MHz): d 8.26 (d, J=7.9 Hz, 4H, H-2pyr), 7.32
d, J=8.2 Hz, 4H, H-2benz), 7.25 (d, J2,3=8.2 Hz, 4H,
2.0 package. PALLAS is a package of powerful tools
for the prediction of certain physicochemical para-
meters of organic compounds based solely on struc-
tural information. PROLOG is an optional component
of the PALLAS system, which predicts the logarithm
of the partition coefficient (log P) of organic com-
pounds in an n-octanol/water system based on chemi-
cal structure.
H-3benz), 7.02 (d, J2,3=7.9 Hz, 4H, H-3pyr), 5.89 (ddt,
J2,3 =17.2, J2,3=10.5, J1,2=4.9 Hz, 4H, H-2allyl), 5.35
s, 4H, CH N ), 5.27 (ddt, J2,3=10.5, J3,3
0
+
(
0
=1.0,
0
=17.2,
2
J1,3=1.0 Hz, 4H, H-3allyl), 5.21 (ddt, J
2,3
0
J3,3
J1,2=4.9, J1,3=1.8, J1,3
J=7.7 Hz, 4H, CH Ph), 1.90 (q, J=7.7 Hz, 2H, C–
0
=1.0, J1,3
0
=1.0 Hz, 4H, H-3 allyl), 4.23 (dt,
0
=1.8 Hz, 4H, H-1allyl), 2.64 (t,
2
1
3
CH –C). C NMR (100.13 MHz): d 157.88 (C-4pyr),
2
Pharmacology
1
44.94 (C-1 ), 143.49 (C-2 ), 133.41 (C-4 ), 131.48
Ph pyr Ph
(
3allyl), 109.90 (C-3pyr), 61.80 (CH N ), 54.23 (C-1allyl),
C-2allyl), 130.50 (C-3 ), 129.60 (C-2 ), 118.35 (C-
+
The ChoK inhibition and anti-proliferative assays
against HT-29 cells were followed in accordance with
the protocols previously reported. In the few cases for
which the two means differed by more than 50%, a third
experiment was performed to ascertain the value.
Ph
Ph
2
6
,7
3
5.99 (CH2 Ph), 34.05 (C–CH –C). HR LSIMS (thio-
2
+
+
glycerol+Na ), calcd m/z for C H N Br (MÀBr)
3
9
46
4
6
49.2906.
Found
m/z:
649.2905.
Anal.
for

J. M. Campos et al. / Bioorg. Med. Chem. 10 (2002) 2215–2231
2231
Acknowledgements
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1
5. Kemp, W., Organic Spectroscopy; MacMillan Education:
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6. Friebolin, H. Basic One- and Two-Dimensional NMR
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7. Schnall, S., Macdonald, J. S. In Manual of Oncologic
Prof. R. B. Silverman’s technical support for the inver-
sion recovery experiments is acknowledged. Funding
´ ´
´
was provided by the Comision de Investigacion Cientı-
´
fica y Tecnica (SAF98-0112-CO2-O1).
1
1
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1
´
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˜
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´
´
1
1
1