Synthesis of pathogen inactivating nucleic acid intercalators

Article abstract of DOI:10.1016/j.ejmech.2004.07.011

A series of antiviral compounds consisting of an intercalating acridine derived part, a spacer region and a reactive EDTA-derived conjugate was synthesized in an easy sequence starting from 1,ω-alkyldiamines. As shown in model screenings, in the presence of ascorbic acid the Fe-complexes of these compounds reduced the phage-titer of MS2-phages by several logarithmic decades.

Full text of DOI:10.1016/j.ejmech.2004.07.011

European Journal of Medicinal Chemistry 39 (2004) 975–988
www.elsevier.com/locate/ejmech
Short communication
Synthesis of pathogen inactivating nucleic acid intercalators
René Csuk *, Alexander Barthel, Thorsten Brezesinski, Christian Raschke
Institut für Organische Chemie, Martin-Luther-Universität Halle-Wittenberg, Kurt-Mothes-Strasse 2, 06120 Halle (Saale), Germany
Received 22 March 2004; received in revised form 19 July 2004; accepted 30 July 2004
Available online 30 September 2004
Abstract
A series of antiviral compounds consisting of an intercalating acridine derived part, a spacer region and a reactive EDTA-derived conjugate
was synthesized in an easy sequence starting from 1,x-alkyldiamines. As shown in model screenings, in the presence of ascorbic acid the
Fe-complexes of these compounds reduced the phage-titer of MS2-phages by several logarithmic decades.
© 2004 Elsevier SAS. All rights reserved.
Keywords: Intercalators; Antiviral agent; Pathogen inactivation; Acridines
1. Introduction
matic units with an annular topology similar to that of DNA
bind in the minor groove in AT-rich regions, the recognition
sequences of regulatory proteins primarily interact in the
major groove usually in GC-rich regions [1,2]. Finally, in a
third way, small molecules may bind to DNA by intercalation
between the base pairs, a mode of reversible binding that is
now understood to be the preferred binding mode of virtually
any flat polyaromatic ligand of sufficiently large surface area
and suitable steric properties [3]. The driving forces for
intercalating are primarily stacking interactions [4–6]
between the ligand chromophore and the base pairs.
A concept for the development of pathogen inactivating
agents might introduce a molecular entity containing an
acridine-derived intercalator [7] that binds to the nucleic acid
of pathogens and of a conjugate that destroys the nucleic
acids subsequently (Fig. 1).
Largely in response to the AIDS epidemic, the amount of
research directed towards the discovery of antiviral agents
has grown dramatically over the last years. Of special interest
have been the detection, the destruction or inactivation of
viruses, bacteria and other pathogens in extracorporal fluids
as blood and plasma. Whereas for the detection and classifi-
cation of bacteria a number of well established methods has
been developed and a repertoire of suitable antibiotics is
readily available, the inactivation of viruses as well as of
other pathogens remains difficult.
This is especially critical in the field of blood transfusion
medicine; progress seems necessary in the development of
techniques for disinfecting donated blood. Such an approach
would offer the major advantage of removing or inactivating
all pathogenic microorganisms from donor blood, even those
for which the blood currently is not—or cannot be—tested.
Various groups are working on inactivation techniques
based on compounds that interact with DNA or RNA of
microorganisms and then bind inextricably with it. As a
result, the microorganisms are no longer able to proliferate
and perish.
There are several generally recognized ways in which
molecules can bind reversibly to DNA or RNA. Whereas
cationic small molecules composed of several repeated aro-
* Corresponding author. Tel.: +49-345-5525660; fax: +49-345-5527030.
E-mail address: csuk@chemie.uni-halle.de (R. Csuk).
Fig. 1. Principal structure of the pathogen inactivating compounds.
0223-5234/$ - see front matter © 2004 Elsevier SAS. All rights reserved.
doi:10.1016/j.ejmech.2004.07.011

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R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
Fig. 2. Suggested Fenton mechanism for the damaging of DNA or RNA.
In our approach, the conjugate consists of a metal–chelate
complex wherein the metal can change between at least two
levels of oxidation. These compounds preferentially show an
acridine [8–10] as the intercalator possessing in addition a
spacer group to an EDTA Fe(II) chelate acting as the conju-
gate. Damage to the DNA should be accomplished by produ-
cing OH radicals by a Fe(II) catalyzed Fenton mechanism
(Fig. 2) [11–13].
Indeed, a variety of products [11–13] may result from free
radical attack [14–20] on DNA, among these the formation of
thymine glycols (from thymine), 4,6-diamino-5-formamido-
pyrimidine (from adenine), 8-hydroxy-guanine (from gua-
nine) and also strand breaks may occur. Addition of a redu-
cing agent such as ascorbic acid leads to a cycle which
increases the damage to the biological molecules.
deesterification using pig liver esterase (PLE)] using the
mixed-anhydride method (isobutyl chloroformate/N-methyl-
morpholine N-oxide (NMM)) furnished the corresponding
triethylesters 33–42. Saponification of the esters was perfor-
med by their treatment with aq. sodium hydroxide to afford
the target compounds 43–52.
The target compounds were treated with an 2–5 molar
excess of Fe3+, lyophilized and incubated with the phages in
Tris-buffer in the presence of sodium ascorbate [13].
3. Results
For biological screening the well established system
containing MS2 bacteriophages (genus of the family levivi-
ridae; uncoated, containing ssRNA) was used; these viruses
contain the short version of the genome and have a separate
gene for cell lysis [35].
2. Chemistry
As shown in Fig. 3 the length of the spacer exhibits
significant effects on the reduction of the phage-titer. Best
results were obtained for a spacer length of n = 6 giving rise
to a reduction of the phage-titer of MS2 phages by > logarith-
mic decades.
As shown for compound 47 (n = 6) the inactivation of this
virus depends both on the concentration and temperature
(Fig. 4), the time of incubation (Fig. 5) as well as on the
concentration of ascorbate (Fig. 6). Increased activity with
increased concentration of ascorbate as well as the observa-
tion that no activity is associated with these compounds in the
lack of ascorbate allows a triggering of the activity by the
addition of ascorbic acid. No significant antipathogenic acti-
vity, however, was found upon incubating the MS2 phages
with acridine, Fe(III)-loaded EDTA in the presence of ascor-
bate.
As far as the synthesis (Scheme 1) of this spacer is concer-
ned, a variety of 1,x-diamines was mono-protected by the
reaction with tert.butyl-dicarbonate in dry dioxane to afford
the corresponding 1-amino-x-tert.butyloxycarbonyl-amino-
alkanes 1 [20], 2 [21], 3 [22], 4 [23], 5 [24], 6 [25], 7 [26], 8
[27], 9 [26], 10 [28]. Thus, usually two equivalents of the
diamine were allowed to react with one equivalent of
tert.butyl-di-carbonate to result in approx. 40% yields of the
corresponding products. These moderate yields could be
improved, however, by using a 5–6 molar excess [29] of the
amine.
These 1-amino-x-tert.butyloxycarbonylamino-alkanes
1–10 were allowed to react with 9-chloro-acridine [30,31] in
the presence of phenol [32,33] to result in the formation of
the corresponding tert.butyl N-[x-(9-acridinyl-amino)alkyl]
carbamates 11–20. Deprotection of these compounds by aq.
hydrochloric acid for 10 h at 30 °C gave the corresponding
dihydrochlorides 21–30. Reaction of these amines with
EDTA-triethylester (31) [34] [that was conveniently prepa-
red even on a larger preparative scale from the corresponding
tetra-ethylester 32 by selective enzymatic mono-
4. Conclusions
In this study we have investigated the antipathogenic acti-
vity of a reactive system constisting of an acridine-derived

R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
977
Scheme 1
NaOH
. (a) (Boc)2O; (b) 9-chloro-acridine/phenol; (c) 10% aq. HCl; (d) EDTA triethylester (31), N-methyl-morpholine, isobutylchloroformate; (e) aq.
Fig. 3. Inactivation of phage MS2 as a function of spacer length (20 °C,
5 mmol Na-ascorbate, 101 µmol of compound 43–52 (Fe3+ loaded).
intercalator, a spacer unit and an EDTA derived reactive part.
Among these derivatives, compound 47 proved to be the
compound showing the highest antipathogenic activity. Al-
though these experiments showed the length of the spacer
being crucial, presently no rationale can be given for this
effect. The inactivation of the virus depends strongly on the
presence of ascorbate obviously to trigger the catalytic
Fig. 4
. Inactivation of MS2 phages as a function of concentration/
temperature (47, incubation time 4 h; 5 mmol Na ascorbate; 25 °C, 37 °C).
Fenton-cycle. Further chemical modification of these com-
pounds and their biological evaluation is underway and these
results will be described in due course.

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R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
5.1.1. General procedure for the synthesis of tert.butyl
N-[x-(9-acridinylamino)alkyl]carbamates (GP1)
A mixture of 9-chloro-acridine (3.3 g, 15.5 mmol) and
phenol (9.0 g, 95.6 mmol) was stirred at 90 °C for 30 min,
then the corresponding 1-amino-x-tert.butyloxycarbonyl
amino-alkane was added and the reaction mixture stirred for
another 30 min at 105 °C. Then all the volatiles were remo-
ved under reduced pressure and the residue subjected to flash
chromatography (silica gel, methanol/ethyl acetate 1:6).
5.1.2. General procedure for the synthesis of the N
(1-(9-acridinyl)-1,x-alkanediamine dihydrochlorides (GP2)
The corresponding tert.butyl N-[x-(9-acridinylamino)
alkyl]carbamate is stirred in methanol (200 ml) and aq.
hydrochloric acid (10%, 20 ml) for 10 h at 30 °C. The
solvents were removed under diminished pressure and the
products were obtained. They were used without any further
purification in the next step.
Fig. 5. Virusinactivation as a function of the incubation time (100 µmol 47,
Na ascorbate 100 µmol, 25 °C).
5.1.3. General procedure for the synthesis of ethyl
2-((2-{[x-(9-acridinylamino)alkyl]amino}-2-oxoethyl)
{2-[di-(2-ethoxy-2-oxoethyl)amino]ethyl} acetates (GP3)
To an ice-cold solution of 31 (1.2 g, 2.67 mmol) and
N-methyl-morpholine (1 ml) in DMF/ethyl acetate (1:1,
50 ml) isobutyl chloroformate (438 mg, 3.2 mmol) is slowly
added via a syringe and the mixture is stirred at this tempera-
ture for 1 h. Then a solution of the corresponding N 1-(9-
acridinyl)-1,2-alkanediamine dihydrochlorides in N-methyl-
morpholine (2 ml) is added and stirring at room temperature is
continued for 24 h. The solvents were removed under dimi-
nished pressure and the residue subjected to chromatography
(silica gel, methanol/ethyl acetate 1:9) to afford the products.
Fig. 6. Virusinactivation as a function of the ascorbate concentration (4 h,
5.1.4. General procedure for the synthesis
25 °C, 50 µg/ml 47).
of the 2-[{2-[(2-{[x-(9-acridinylamino)alkyl]amino}-
2-oxo-ethyl) (carboxymethyl)amino]ethyl}(carboxymethyl)
amino] acetic acids (GP4)
5. Experimental
The corresponding ethyl 2-((2-{[x-(9-acridinylamino)
alkyl]amino}-2-oxoethyl){2-[di-(2-ethoxy-2-oxo-ethyl)ami-
no]ethyl}amino) acetate and sodium hydroxide are dissolved
in ethanol/water (5:1, 40 ml) and stirred at room temperature
for 12 h; then the pH was carefully adjusted to 7. The solvents
were removed under diminished pressure and the residue was
carefully washed with ice-cold water (20 ml) and chloroform
(150 ml) to obtain the products.
5.1. General
Melting points are uncorrected (Leica hot stage micros-
cope), NMR spectra were recorded using the Varian spectro-
meters Gemini 200, Gemini 2000 or Unity 500 (d given in
ppm, J in Hz, internal Me4Si), IR spectra (film or KBr pellet)
on a Perkin-Elmer FT-IR spectrometer Spectrum 1000, MS
spectra were taken on a Intectra GmbH AMD 402 (electron
impact, 70 eV) or on a Finnigan MAT TSQ 7000 (electros-
pray, voltage 4.5 kV, sheath gas nitrogen) instrument; for
elemental analysis a Foss-Heraeus Vario EL instrument was
used; TLC was performed on silica gel (Merck 5554, detec-
tion by UV absorption or by treatment with a solution of 10%
sulfuric acid, ammonium molybdate and cerium(IV)) sulfate
followed by gentle heating. The solvents were dried accor-
ding to usual procedures. Analyses indicated by the symbols
of the elements were within 0.3% of the theoretical values.
5.2. Bacteriophages and screening
The effect of the compounds on bacteriophages was tested
on bacteriophages MS2 grown on its E. coli ATCC15597
host. Log-phase host bacteria for bacteriophage propagation
were grown in tryptic soy broth (TSB, Sigma Chemicals) on
an orbital shaker at room temperature until turbid. Before
use, 100 µl of culture was inoculated into fresh TSB contai-
ning 0.0025% CaCl2. The cultures were incubated in a 37 °C
shaking water bath for 4 h until the log-phase was reached.

R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
979
Fresh bacteriophages were cultured from freezer stocks.
Log-phase host bacteria and phages were mixed at a multi-
plicity of infection of approximately 1 in 5 ml TSB. The
culture was kept on ice for 15 min to facilitate adsorption of
the phages to the host cells, then incubated overnight at
37 °C, the phage culture was filtered through a 0.2 µm
cellulose acetate syringe filter to remove host bacteria and
then stored at 4 °C. Typical yields were 1 × 1010 PFU/ml.
Culturable counts of phage were performed by mixing 100 µl
of phage suspension and 100 µl of host culture in 4 ml molten
TSB top agar (containing 0.7% agar). The top agar was
vortexed gently, then poured on TSB plates; the plates were
incubated at 37 °C. Bacteriophages from freeze stocks were
diluted into buffer (30 mM Tris, 150 mM KCl, pH 8.3) to a
final population density of approximately 1 × 109 PFU/ml.
Dilutions were prepared in phosphate-buffered saline solu-
tion. Initial as well post-exposure culturable counts were
performed in triplicate. The counts were divided by the mean
unexposed control counts to normalize the data and then
log-transformed.
1 H, NH), 3.81–3.78 (m, 2 H, CH2(1′)), 3.38–3.36 (m, 2 H,
CH2(3′)), 1.90–1.84 (m, 2 H, CH2(2′)), 1.47 (s, 9 H,tBu); 13
C
NMR (100 MHz, CDCl3): d = 156.9 (C=O), 151.9 (quart.,
C9), 148.8 (quart.), 129.8 (CH), 128.9 (CH), 123.1 (CH),
122.9 (CH), 117.1 (quart.), 79.8 (quart., tBu), 47.1 (CH2(1′)),
t
37.7 (CH2(3′)), 32.2 (CH2(2′)), 28.5 (CH3, Bu); MS (ESI,
4.1 kV, 8 µl/min, N2, methanol): m/z = 252 (2%) [(M-
Boc)H]+, 296 (8%) [(M-ibutene)H]+, 352 (100%) [MH]+;
HRMS (C21H25N3O2): 351.19466; found: 351.19467; Anal.
C21H25N3O2 (C, H, N).
5.2.3. Tert.butyl N-[4-(9-acridinylamino)butyl]carbamate
(13)
Following GP1 from 9-chloro-acridine (3.0 g,
14.0 mmol), phenol (8.0 g, 85.0 mmol) and 3 (2.8 g,
15.0 mmol) 13 (4.3 g, 84%) was obtained as an amorphous,
orange-coloured solid; RF (methanol/ethyl acetate 1:3) 0.61;
UV-vis (methanol): kmax (log e) = 283 nm (4.76); IR (KBr):
m = 2930w, 1688s, 1636m, 1590s, 1562s, 1520s, 1472m,
1
1391m, 1253s, 1170s, 858w, 756s, 651w cm–1; H NMR
3
(400 MHz, CDCl3): d = 8.10 (d, JH,H = 8.72 Hz, 2 H,
H-C(4,5)), 8.00 (d, 3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.55 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.28 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.69 (br s,
1 H, NH), 3.92–3.89 (m, 2 H, CH2(1′)), 3.24–3.14 (m, 2 H,
CH2(4′)), 1.94–1.84 (m, 2 H, CH2(2′)), 1.71–1.63 (m, 2 H,
CH2(3′)), 1.42 (s, 9 H, tBu); 13C NMR (100 MHz, CDCl3):
d = 156.1 (C=O), 153.1 (quart., C9), 145.8 (quart.), 131.1
(CH), 125.8 (CH), 123.6 (CH), 122.9 (CH), 115.2 (quart.),
5.2.1. Tert.butyl N-[2-(9-acridinylamino)ethyl]carbamate
(11)
Following GP1 from 9-chloro-acridine (3.3 g,
15.5 mmol), phenol (9 g, 95.6 mmol) and 1 (2.7 g, 17 mmol)
11 (4.5 g, 87%) was obtained as an amorphous, red solid. RF
(methanol/ethyl acetate 1:3) 0.62; UV-vis (methanol): kmax
(log e) = 283 nm (4.81); IR (KBr): m = 2978m, 1687s, 1636m,
1561s, 1522s, 1261m, 1167m, 857w, 758m, 652w cm–1; 1H
NMR (500 MHz, CDCl3): d = 8.16 (d, 3JH,H = 8.72 Hz, 2 H,
H-C(4,5)), 7.98 (d, 3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.53 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.24 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 5.50 (br s,
1 H, NH), 4.00–3.99 (m, 2 H, CH2(1′)), 3.61–3.60 (m, 2 H,
CH2(2′)), 1.44 (s, 9 H,tBu); 13C NMR (125 MHz, CDCl3):
d = 157.8 (C=O), 152.8 (quart., C9), 146.6 (quart.), 130.8
(CH), 126.3 (CH), 123.7 (CH), 122.7 (CH), 115.1 (quart.),
80.3 (quart., tBu), 52.2 (CH2(1′)), 41.0 (CH2(2′)), 28.4 (CH3,
tBu); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z = 238
(4%) [(M-Boc)H]+, 282 (64%) [(M-ibutene)H]+, 338 (100%)
[MH]+; HRMS (C20H23N3O2): 337.17901; found:
337.17903; Anal. C20H23N3O2 (C, H, N).
t
79.3 (quart., Bu), 49.8 (CH2(1′)), 40.1 (CH2(4′)), 28.5
t
(CH2), 28.4 (CH3, Bu), 27.7 (CH2); MS (ESI, 4.1 kV,
8 µl/min, N2, methanol): m/z = 266 (1%) [(M-Boc)H]+, 310
(9%) [(M-ibutene)H]+, 366 (100%) [MH]+; HRMS
(C22H27N3O2): 365.2101; found: 365.2103; Anal.
C22H27N3O2 (C, H, N).
5.2.4. Tert.butyl N-[5-(9-acridinylamino)pentyl]carbamate
(14)
Following GP1 from 9-chloro-acridine (4.0 g,
18.7 mmol), phenol (10.0 g, 0.11 mol) and 4 (4.2 g,
20.6 mmol) 14 (5.7 g, 80.0%) was obtained as an amorphous,
orange-coloured solid; RF (methanol/ethyl acetate 1:3) 0.61;
UV-vis (methanol): kmax (log e) = 284 nm (4.73); IR (KBr):
m = 2979w, 1683s, 1634s, 1594s, 1533s, 1474w, 1366m,
5.2.2. Tert.butyl N-[3-(9-acridinylamino)propyl]
carbamate (12)
1
1272m, 1171m, 756w cm–1; H NMR (400 MHz, CDCl3):
3
Following GP1 from 9-chloro-acridine (3.5 g,
16.4 mmol), phenol (9.0 g, 95.6 mmol) and 2 (3.14 g,
18.0 mmol) 12 (5.5 g, 95.5%) was obtained as an amorphous
solid. RF (methanol/ethyl acetate 1:3) 0.60; UV-vis (metha-
nol): kmax (log e) = 282 nm (4.77); IR (KBr): m = 2976w,
2935w, 1683s, 1616m, 1564s, 1530s, 1393m, 1290m,
1263m, 1167m, 1141m, 758m, 647w cm–1; 1H NMR
d = 8.20 (d, JH,H = 8.72 Hz, 2 H, H-C(4,5)), 7.93 (d,
3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.38 (dd, 3JH,H = 8.72 Hz,
3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.14 (dd, 3JH,H = 8.72 Hz,
3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.75 (br t, 3JH,H = 5.40 Hz,
1 H, NH), 4.07–4.04 (m, 2 H, CH2(1′)), 3.18–3.13 (m, 2 H,
CH2(5′)), 2.11–2.01 (m, 2 H, CH2(2′)), 1.67–1.51 (m, 4 H,
2 × CH2(3′,4′)), 1.40 (s, 9 H,tBu); 13C NMR (100 MHz,
CDCl3): d = 156.2 (C=O), 156.0 (quart., C9), 140.2 (quart.),
133.2 (CH), 125.0 (CH), 122.8 (CH), 120.0 (CH), 112.5
3
(400 MHz, CDCl3): d = 8.19 (d, JH,H = 8.72 Hz, 2 H,
H-C(4,5)), 8.02 (d, 3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.62 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.32 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.87f (br s,
t
(quart.), 79.1 (quart., Bu), 48.5 (CH2(1′)), 40.2 (CH2(5′)),
t
30.0 (CH2), 29.8 (CH2), 28.5 (CH3, Bu), 24.2 (CH2); MS

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R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
(ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z = 280 (2%)
[(M-Boc)H]+, 324 (7%) [(M-ibutene)H]+, 380 (100%)
[MH]+; HRMS (C23H29N3O2): 379.22596; found: C
379.22599; Anal. C23H29N3O2 (C, H, N).
5.2.7. Tert.butyl N-[8-(9-acridinylamino)octyl]carbamate
(17)
Following GP1 from 9-chloro-acridine (1.4 g, 6.4 mmol),
phenol (5.0 g, 53.1 mmol) and 7 (1.8 g, 7.36 mmol) 17 (2.2 g,
81.5%) was obtained as an amorphous, orange-coloured so-
lid; RF (methanol/ethyl acetate 1:3) 0.67; UV-vis (methanol):
kmax (log e) = 284 nm (4.76); IR (KBr): m = 2931w, 1635s,
1594s, 1570s, 1537s, 1469s, 1257m, 1173m, 859w, 750m,
5.2.5. Tert.butyl N-[6-(9-acridinylamino)hexyl] carbamate
(15)
Following GP1 from 9-chloro-acridine (3.0 g,
14.0 mmol), phenol (7.0 g, 74.4 mmol) and 5 (3.2 g,
15.0 mmol) 15 (5.0 g, 91%) was obtained as an orange-
coloured amorphous solid; RF (methanol/ethyl acetate 1:3)
0.63; UV-vis (methanol): kmax (log e) = 281 nm (4.63); IR
(KBr): m = 2932w, 1687s, 1634s, 1570s, 1533s, 1474s,
1391m, 1273m, 1252m, 1171s, 860w, 756m, 661w cm–1; 1H
NMR (400 MHz, CDCl3): d = 8.07 (d, 3JH,H = 8.72 Hz, 2 H,
H-C(4,5)), 7.95 (d, 3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.55 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.27 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.55 (br s,
1 H, NH), 3.86–3.82 (m, 2 H, CH2(1′)), 3.11–3.04 (m, 2 H,
CH2(6′)), 1.87–1.78 (m, 2 H, CH2(2′)), 1.50–1.42 (m, 4 H,
2 × CH2(3′,5′)), 1.41 (s, 9 H,tBu), 1.40–1.36 (m, 2 H,
CH2(4′)); 13C NMR (125 MHz, CDCl3): d = 156.3 (C=O),
156.1 (quart., C9), 140.5 (quart.), 133.4 (CH), 125.1 (CH),
1
660w cm–1; H NMR (400 MHz, CDCl3): d = 8.21 (d,
3
3JH,H = 8.72 Hz, 2 H, H-C(4,5)), 7.93 (d, JH,H = 8.72 Hz,
2 H, H-C(1,8)), 7.35–7.34 (m, 2 H, H-C(3,6)), 7.12 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.55 (br s,
1 H, NH), 4.08–4.04 (m, 2 H, CH2(1′)), 3.09–3.03 (m, 2 H,
CH2(8′)), 2.07–1.99 (m, 2 H, CH2(2′)), 1.56–1.40 (m, 6 H,
3 × CH2(3′,6′,7′)), 1.41 (s, 9 H,tBu), 1.36–1.24 (m, 4 H, 2 ×
CH2(4′,5′)); 13C NMR (100 MHz, CDCl3): d = 156.6 (C=O),
155.9 (quart., C9), 139.6 (quart.), 133.5 (CH), 125.2 (CH),
t
122.7 (CH), 119.3 (CH), 112.2 (quart.), 79.0 (quart., Bu),
48.4 (CH2(1′)), 40.7 (CH2(8′)), 30.4 (CH2), 30.1 (CH2),
29.23 (CH2), 29.18 (CH2), 28.5 (CH3, tBu), 27.0 (CH2), 26.8
(CH2); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z = 322
(5%) [(M-Boc)H]+, 366 (10%) [(M-ibutene)H]+, 422 (100%)
[MH]+; HRMS (C26H35N3O2): 421.27291; found:
421.27294; Anal. C26H35N3O2 (C, H, N).
t
122.9 (CH), 120.2 (CH), 112.6 (quart.), 79.0 (quart., Bu),
48.4 (CH2(1′)), 40.2 (CH2(6′)), 30.2 (CH2), 29.9 (CH2), 28.4
t
(CH3, Bu), 26.3 (CH2), 26.2 (CH2); MS (ESI, 4.1 kV,
5.2.8. Tert.butyl N-[9-(9-acridinylamino)nonyl] carbamate
(18)
8 µl/min, N2, methanol): m/z = 294 (3%) [(M-Boc)H]+, 338
(7%) [(M-ibutene)H]+, 394 (100%) [MH]+; HRMS
(C14 H31N3O2): 393.24161; found: 393.24162; Anal.
C14H31N3O2 (C, H, N).
Following GP1 from 9-chloro-acridine (2.33 g,
11.0 mmol), phenol (8.0 g, 85 mmol) and 8 (3.1 g,
12.0 mmol) 18 (4.4 g, 92%) was obtained as a red, amor-
phous solid; RF (methanol/ethyl acetate 1:3) 0.69; UV-vis
(methanol): kmax (log e) = 284 m (4.73); IR (KBr):
m = 2926w, 1687s, 1633m, 1561m, 1521m, 1471m, 1251s,
5.2.6. Tert.butyl N-[7-(9-acridinylamino)heptyl] carbamate
(16)
1
Following GP1 from 9-chloro-acridine (3.3 g,
15.4 mmol), phenol (9.0 g, 95.6 mmol) and 6 (3.9 g,
17.0 mmol) 16 (6.3 g, 83%) was obtained as an amorphous,
orange-coloured solid; RF (methanol/ethyl acetate 1:3) 0.64;
UV-vis (methanol): kmax (log e) = 284 nm (4.72); IR (KBr):
m = 2929w, 1691s, 1635m, 1563m, 1522m, 1473m, 1391m,
1169m, 756m, 651w cm–1; H NMR (400 MHz, CDCl3):
3
d = 8.12 (d, JH,H = 8.72 Hz, 2 H, H-C(4,5)), 7.98 (d,
3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.49 (dd, 3JH,H = 8.72 Hz,
3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.22 (dd, 3JH,H = 8.72 Hz,
3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.51 (br s, 1 H, NH),
3.96–3.92 (m, 2 H, CH2(1″)), 3.10–3.02 (m, 2 H, CH2(9′)),
1.94–1.85 (m, 2 H, CH2(2′)), 1.50–1.42 (m, 4 H, 2 ×
CH2(3′,8′)), 1.42 (s, 9 H,tBu), 1.42–1.23 (m, 8 H, 4 ×
CH2(4′,5′,6′,7′); 13C NMR (100 MHz, CDCl3): d = 155.9
(C=O), 154.1 (quart., C9), 144.0 (quart.), 131.7 (CH), 124.0
1272m, 1170m, 754m, 652w cm–1; H NMR (400 MHz,
1
CDCl3): d = 8.08 (d, 3JH,H = 8.72 Hz, 2 H, H-C(4,5)), 7.99 (d,
3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.57 (dd, 3JH,H = 8.72 Hz,
3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.28 (dd, 3JH,H = 8.72 Hz,
3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.51 (br s, 1 H, NH),
3.87–3.83 (m, 2 H, CH2(1′)), 3.09–3.04 (m, 2 H, CH2(7′)),
1.86–1.79 (m, 2 H, CH2(2′)), 1.47–1.41 (m, 4 H, 2 ×
CH2(3′,6′)), 1.41 (s, 9 H,tBu), 1.40–1.27 (m, 4 H, 2 ×
CH2(4′,5′)); 13C NMR (100 MHz, CDCl3): d = 155.9 (C=O),
154.6 (quart., C9), 140.8 (quart.), 132.1 (CH), 124.2 (CH),
t
(CH), 122.8 (CH), 114.1 (quart.), 79.0 (quart., Bu), 49.6
(CH2(1′)), 40.6 (CH2(9′)), 31.0 (CH2), 30.1 (CH2), 29.4
(CH2), 29.2 (CH2), 29.1 (CH2), 28.5 (CH3, tBu), 26.9 (CH2),
26.7 (CH2); MS (ESI, 4.1 V, 8 µl/min, N2, methanol):
m/z = 336 (7%) [(M-Boc)H]+, 380 (13%) [(M-ibutene)H]+,
436 (100%) [MH]+; HRMS (C27H37N3O2): 435.28856;
found: 435.28857; Anal. C27H37N3O2 (C, H, N).
t
122.8 (CH), 117.0 (CH), 113.7 (quart.), 79.0 (quart., Bu),
49.2 (CH2(1′)), 40.5 (CH2(7′)), 30.8 (CH2), 30.0 (CH2), 28.9
t
5.2.9. Tert.butyl N-[10-(9-acridinylamino)decyl]
carbamate (19)
According to GP1 from 9-chloro-acridine (3.0 g,
14 mmol), phenol (9.0 g, 95.6 mmol) and 9 (3.5 g,
13.0 mmol) followed by chromatography (silica gel,
(CH2), 28.5 (CH3, Bu), 26.8 (CH2), 26.6 (CH2); MS (ESI,
4.1 kV, 8 µl/min, N2, methanol): m/z = 308 (7%) [(M-
Boc)H]+, 352 (11%) [(M-ibutene)H]+, 408 (100%) [MH]+;
HRMS (C25H33N3O2): 407.25726; found: 407.25728; Anal.
C25H33N3O2 (C, H, N).

R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
981
methanol/ethylacetate 1:4 then methanol/ethyl acetate 1:6)
19 (3.6 g, 62%) was obtained as an orange-coloured, amor-
phous solid; RF (methanol/ethyl acetate 1:3) 0.70; UV-vis
(methanol): kmax (log e) = 284 nm (4.93); IR (KBr):
m = 2927m, 1691s, 1615s, 1560s, 1520m, 1473m, 1391m,
1258m, 1172m, 857w, 757m, 650w cm–1; 1H NMR
m = 1638s, 1588s, 1527s, 1469m, 1339m, 1270m, 1176w,
868w, 817w, 755m, 670w cm–1; 1H NMR (400 MHz, D2O):
3
d = 8.10 (d, JH,3H = 8.72 Hz, 2 H, H-C(4,5)), 7.79 (dd,
3JH,H = 8.72 Hz, JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.52 (d,
3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.40 (dd, 3JH,H = 8.72 Hz,
3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.27–4.24 (m, 2 H, CH2(1′)),
3.41–3.38 (m, 2 H, CH2(2′)); 13C NMR (100 MHz, D2O):
d = 156.0 (quart.), 139.3 (quart.), 135.4 (CH), 124.5 (CH),
124.0 (CH), 118.4 (CH), 112.1 (quart.), 45.5 (CH2(1′)), 38.5
(CH2(2′)); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol):
m/z = 238 (100%) [MH]+.
3
(400 MHz, CDCl3): d =8.07 (d, JH,H = 8.72 Hz, 2 H,
H-C(4,5)), 8.05 (d, 3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.65 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.35 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 5.07 (br s,
1 H, NH) 4.50 (br s, 1 H, NH), 3.82–3.78 (m, 2 H, CH2(1′)),
3.11–3.03 (m, 2 H, CH2(10′)), 1.80–1.73 (m, 2 H, CH2(2′)),
1.42–1.40 (m, 13 H,tBu, 2 × CH2(3′,9′)), 1.31–1.23 (m, 10 H,
5 × CH2(4′,5′,6′,7′,8′); 13C NMR (100 MHz, CDCl3):
d = 155.9 (C=O), 151.2 (quart., C9), 149.3 (quart.), 129.7
(CH), 129.6 (br, CH), 122.9 (CH), 122.6 (CH), 116.6
(quart.), 79.0 (quart., tBu), 51.0 (CH2(1′)), 40.7 (CH2(10′)),
31.9 (CH2), 30.1 (CH2), 29.4 (CH2), 29.3 (CH2), 29.2 (CH2),
28.5 (CH3, tBu), 26.9 (CH2), 26.8 (CH2); MS (ESI, 4.1 kV,
8 µl/min, N2, methanol): m/z = 350 (8%) [(M-Boc)H]+, 394
(11%) [(M-ibutene)H]+, 450 (100%) [MH]+; HRMS
(C28H39N3O2): 449.30421; found: 449.30423; Anal.
C28H39N3O2 (C, H, N).
5.2.12. N (1-(9-Acridinyl)-1,3-propanediamine
dihydrochloride (22)
Following GP2 from 12 (5.5 g, 15.67 mmol) 22 (4.9 g,
96%) was obtained as an orange-coloured amorphous solid;
UV-vis (methanol): kmax (log e) = 282 nm (4.75); IR (KBr):
m = 3071m, 3030m, 2992m, 1635s, 1592s, 1575s, 1532s,
1476s, 1336m, 1276m, 1172m, 852w, 801w, 756m, 662m
1
cm–1; H NMR (400 MHz, D2O): d = 7.85–7.75 (m, 2 H,
H-C(4,5)), 7.67–7.57 (m, 2 H, H-C(3,6)), 7.29–7.24 (m, 2 H,
H-C(1,8)), 7.16–7.13 (m, 2 H, H-C(2,7)), 3.80–3.75 (m, 2 H,
CH2(1′)), 2.98–2.92 (m, 2 H, CH2(3′)), 2.07–2.00 (m, 2 H,
CH2(2′)); 13C NMR (100 MHz, D2O): d = 156.5 (quart.),
138.5 (quart.), 135.3 (CH), 124.4 (CH), 124.0 (CH), 118.0
(CH), 111.4 (quart.), 45.8 (CH2(1′)), 37.0 (CH2(3′)), 27.2
(CH2(2′)); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol):
m/z = 252 (100%) [MH]+.
5.2.10. Tert.butyl N-[12-(9-acridinylamino)dodecyl]
carbamate (20)
Following the procedure given for the synthesis of 19
from 9-chloro-acridine (2.4 g, 11.0 mmol), phenol (10.0 g,
0.11 mol) and 10 (3.6 g, 12.0 mmol) 20 (3.4 g, 65%) was
obtained as a red, amorphous solid; RF (methanol/ethyl ace-
tate 1:3) 0.73; UV-vis (methanol): kmax (log e) = 282 nm
(4.70); IR (KBr): m = 2925m, 1694s, 1635s, 1589s, 1568s,
1532s, 1472s, 1390m, 1251m, 1170s, 860w, 754m, 661w
cm–1; 1H NMR (400 MHz, CDCl3): d = 8.18 (d,
5.2.13. N (1-(9-Acridinyl)-1,4-butanediamine
dihydrochloride (23)
Following GP2 from 13 (4.3 g, 11.78 mmol) 23 (3.8 g,
95%) was obtained as a orange-coloured, amorphous solid;
UV-vis (methanol): kmax (log e) = 283 nm (4.76); IR (KBr):
m = 2928m, 1635s, 1592s, 1571m, 1535w, 1474m, 1346w,
3
3JH,H = 8.72 Hz, 2 H, H-C(4,5)), 7.94 (d, JH,H = 8.72 Hz,
1
2 H, H-C(1,8)), 7.41–7.37 (m, 2 H, H-C(3,6)), 7.14 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.49 (br s,
1 H, NH), 4.05–4.01 (m, 2 H, CH2(1′)), 3.09–3.05 (m, 2 H,
CH2(12′)), 2.04–1.97 (m, 2 H, CH2(2′)), 1.57–1.45 (m, 2 H,
CH2(11′)), 1.45–1.41 (m, 13 H,tBu, 2 × CH2(3′,10′)), 1.34–
1.21 (m, 12 H, 6 × CH2(4′,5′,6′,7′,8′,9′); 13C NMR
(100.6 MHz, CDCl3): d = 156.6 (C=O), 141.0 (quart.), 133.0
(CH), 124.9 (CH), 122.8 (CH), 120.7 (CH), 112.9 (quart.),
1271w, 1174w, 859w, 802w, 755m, 663w cm–1; H NMR
3
(400 MHz, D2O): d = 7.78 (d, JH,H = 8.72 Hz, 2 H,2 ×
CH(4,5)), 7.61 (dd, 3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, 2 ×
CH(3,6)), 7.22 (dd, 3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H,2 ×
CH(2,7)), 7.16 (d, 3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 3.68–3.65
(m, 2 H, CH2(1′)), 2.91–2.87 (m, 2 H, CH2(4′)), 1.75–1.66
(m, 2 H, CH2(2′)), 1.64–1.56 (m, 2 H, CH2(3′)); 13C NMR
(125 MHz, D2O): d = 156.2 (quart.), 138.3 (quart.), 135.3
(CH), 124.05 (CH), 124.03 (CH), 118.0 (CH), 111.2 (quart.),
46.2 (CH2(1′)), 39.3 (CH2(4′)), 26.4 (CH2), 24.4 (CH2); MS
(ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z = 266 (100%)
[MH]+.
t
79.0 (quart., Bu), 48.9 (CH2(1′)), 40.7 (CH2(12′)), 30.6
(CH2), 30.2 (CH2), 29.6 (CH2), 29.4 (CH2), 29.3 (CH2), 28.5
t
(CH3, Bu), 27.0 (CH2), 26.9 (CH2); MS (ESI, 4.1 kV,
8 µl/min, N2, methanol): m/z = 378 (6%) [(M-Boc)H]+, 422
(10%) [(M-ibutene)H]+, 478 (100%) [MH]+; HRMS
(C30H43N3O2): 477.33551; found: 477.33553; Anal.
C30H43N3O2 (C, H, N).
5.2.14. N (1-(9-Acridinyl)-1,5-pentanediamine
dihydrochloride (24)
Following GP2 from 14 (5.7 g, 15.04 mmol) 24 (5.1 g,
96%) was obtained as an orange-coloured, amorphous solid;
UV-vis (methanol): kmax (log e) = 284 nm (4.78); IR (KBr):
m = 2928m, 1634s, 1592s, 1570m, 1532w, 1472m, 1340w,
1272w, 1171w, 859w, 754m, 661w cm–1; 1H NMR
5.2.11. N (1-(9-Acridinyl)-1,2-ethanediamine
dihydrochloride (21)
Following GP2 from 11 (4.5 g, 13.35 mmol) 21 (4.03 g,
97%) was obtained as an amorphous, yellow solid. UV-vis
(methanol): kmax (log e) = 283 nm (4.69); IR (KBr):
3
(400 MHz, D2O): d = 7.92 (d, JH,H = 8.72 Hz, 2 H,

982
R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
H-C(4,5)), 7.69 (dd, 3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H,
H-C(3,6)), 7.32–7.27 (m, 4 H, H-C(1,8), H-C(2,7)), 3.77–
3.73 (m, 2 H, CH2(1′)), 2.86–2.80 (m, 2 H, CH2(5′)), 1.75–
1.68 (m, 2 H, CH2(2′)), 1.59–1.52 (m, 2 H, CH2(4′)), 1.36–
1.28 (m, 2 H, CH2(3′)); 13C NMR (125 MHz, D2O): d = 156.9
(quart.), 135.0 (CH), 123.6 (br, CH), 117.9 (CH), 48.4
(CH2(1′)), 39.2 (CH2(5′)), 28.5 (CH2), 26.4 (CH2), 23.0
(CH2); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z = 280
(100%) [MH]+.
CH2(8′)), 1.60–1.58 (m, 2 H, CH2(2′)), 1.48–1.44 (m, 2 H,
CH2(7′)), 1.16 (br s, 8 H, 4 × CH2(3′,4′,5′,6′)); 13C NMR
(100 MHz, D2O): d = 156.8 (quart.), 135.1 (CH), 123.7 (CH),
118.0 (CH), 48.8 (CH2(1′)), 39.6 (CH2(8′)), 29.1 (CH2), 28.2
(CH2), 28.2 (CH2), 26.8 (CH2), 25.9 (CH2), 25.6 (CH2); MS
(ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z = 322 (100%)
[MH]+.
5.2.18. N (1-(9-Acridinyl)-1,9-nonanediamine
dihydrochloride (28)
5.2.15. N (1-(9-Acridinyl)-1,6-hexanediamine
dihydrochloride (25)
Following GP2 from 18 (4.4 g, 10.12 mmol) 28 (4.0 g,
97%) was obtained as a yellow, amorphous solid; UV-vis
(methanol): kmax (log e) = 284 nm (4.67); IR (KBr):
m = 2931m, 1634s, 1589s, 1568s, 1540m, 1478s, 1396m,
1362m, 1339m, 1274m, 1191m, 1168m, 858w, 759m, 657w
Following GP2 from 15 (5.0 g, 12.72 mmol) 25 (4.5 g,
97%) was obtained as an amorphous, yellow solid; UV-vis
(methanol): kmax (log e) = 283 nm (4.52); IR (KBr):
m = 1634s, 1588s, 1538m, 1471s, 1335m, 1269w, 1221w,
872w, 757m, 678w cm–1; 1H NMR (400 MHz, D2O):
cm–1; 1H NMR (400 MHz, D2O):
d = 7.79 (d,
3JH,H = 8.92 Hz, 2 H, H-C(4,5)), 7.62 (t, 3JH,H = 7.26 Hz, 2 H,
H-C(3,6)), 7.21 (d, 3JH,H = 8.51 Hz, 2 H, H-C(1,8)), 7.20 (m,
2 H, H-C(2,7)), 3.63–3.59 (m, 2 H, CH2(1′)), 2.82–2.78 (m,
2 H, CH2(9′)), 1.64–1.56 (m, 2 H, CH2(2′)), 1.49–1.42 (m,
2 H, CH2(8′)), 1.20–1.08 (m, 10 H, 5 × CH2(3′,4′,5′,6′,7′));
13C NMR (100.6 MHz, D2O): d = 156.5 (quart.), 135.0 (CH),
117.9 (CH), 48.7 (CH2(1′)), 39.6 (CH2(9′)), 29.0 (CH2), 28.5
(CH2), 28.2 (CH2), 28.2 (CH2), 26.8 (CH2), 26.0 (CH2), 25.6
(CH2); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z = 168
(18%) [MH2]2+, 336 (100%) [MH]+.
3
d = 7.59 (d, JH,H = 8.72 Hz, 2 H, H-C(4,5)), 7.49 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.10 (dd,
3
3JH,H = 8.72 Hz, JH,H = 7.47 Hz, 2 H, H-C(2,7)), 6.98 (d,
3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 3.44–3.40 (m, 2 H, CH2(1′)),
2.86–2.82 (m, 2 H, CH2(6′)), 1.56–1.48 (m, 4 H, CH2(2′,5′)),
1.28–1.23 (m, 4 H, CH2(3′,4′)); 13C NMR (100 MHz, D2O):
d = 155.9 (quart.), 135.1 (CH), 123.8 (CH), 117.8 (CH), 48.6
(CH2(1′)), 39.6 (CH2(6′)), 29.1 (CH2), 26.9 (CH2), 25.7
(CH2), 25.5 (CH2); MS (ESI, 4.1 kV, 8 µl/min, N2, metha-
nol): m/z = 294 (100%) [MH]+.
5.2.19. N (1-(9-Acridinyl)-1,10-decanediamine
dihydrochloride (29)
5.2.16. N (1-(9-Acridinyl)-1,7-heptanediamine
dihydrochloride (26)
Following GP2 from 19 (3.6 g, 8.02 mmol) 29 (3.3 g,
98%) was obtained as an orange-coloured, amorphous solid;
UV-vis (methanol): kmax (log e) = 281 nm (4.61); IR (KBr):
m = 2924m, 1634s, 1589s, 1531s, 1468s, 1339m, 1271m,
Following GP2 from 16 (6.3 g, 15.48 mmol) 26 (5.7 g,
97%) was obtained as a yellow, amorphous solid; UV-vis
(methanol): kmax (log e) = 284 nm (4.76); IR (KBr):
m = 2931m, 1634s, 1588s, 1567s, 1531s, 1473m, 1376w,
1
1190m, 1169w, 858w, 754m cm–1; H NMR (400 MHz,
1335w, 1270w, 1176w, 879w, 748m, 669w cm–1; H NMR
D2O): d = 7.60 (m, 3JH,H = 8.71 Hz, 2 H, H-C(4,5)), 7.60 (m,
2 H, H-C(3,6)), 7.19 (m, 4 H, H-C(1,8), H-C(2,7)), 3.59–3.55
(m, 2 H, CH2(1′)), 2.83–2.79 (m, 2 H, CH2(10′)), 1.59–1.56
(m, 2 H, CH2(2′)), 1.48–1.42 (m, 2 H, CH2(9′)), 1.08–1.16
(m, 12 H, 6 × CH2(3′,4′,5′,6′,7′,8′)); 13C NMR (100 MHz,
CD3OD): d = 159.5 (quart.), 136.3 (CH), 129.7 (CH), 126.1
(CH), 119.5 (CH), 48.4 (CH2(1′)), 40.8 (CH2(11′)), 30.7
(CH2), 30.5 (CH2), 30.4 (CH2), 30.3 (CH2), 30.2 (CH2), 28.6
(CH2), 27.9 (CH2), 27.5 (CH2); MS (ESI, 4.1 kV, 8 µl/min,
N2, methanol): m/z = 175 (45%) [MH2]2+, 350 (100%)
[MH]+.
1
3
(400 MHz, D2O): d = 7.59 (d, JH,H = 8.51 Hz, 2 H,
H-C(4,5)), 7.50 (t, 3JH,H = 7.26 Hz, 2 H, H-C(3,6)), 7.09 (t,
3
3JH,H = 7.57 Hz, 2 H, H-C(2,7)), 7.00 (d, JH,H = 8.51 Hz,
2 H, H-C(1,8)), 3.43–3.39 (m, 2 H, CH2(1′)), 2.85–2.81 (m,
2 H, CH2(7′)), 1.52–1.48 (m, 4 H, 2 × CH2(2′,6′)), 1.20 (br s,
6 H, 3 × CH2(3′,4′,5′)); 13C NMR (100 MHz, D2O): d = 155.2
(quart.), 134.9 (CH), 123.8 (CH), 117.6 (CH), 48.4
(CH2(1′)), 39.6 (CH2(7′)), 29.1 (CH2), 27.9 (CH2), 26.8
(CH2), 25.9 (CH2), 25.7 (CH2); MS (ESI, 4.1 kV, 8 µl/min,
N2, methanol): m/z = 308 (100%) [MH]+.
5.2.17. N (1-(9-Acridinyl)-1,8-octanediamine
dihydrochloride (27)
5.2.20. N (1-(9-Acridinyl)-1,12-dodecanediamine
dihydrochloride (30)
Following GP2 from 17 (2.2 g, 5.23 mmol) 27 (1.9 g,
93%) was obtained as a yellow, amorphous solid; UV-vis
(methanol): kmax (log e) = 284 nm (4.66); IR (KBr):
m = 1634s, 1588s, 1540s, 1470s, 1363m, 1337m, 1271m,
1190m, 1166m, 869w, 753s, 670w cm–1; 1H NMR
(400 MHz, D2O): d = 7.77 (m, 2 H, H-C(4,5)), 7.61 (t,
3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.20 (m, 4 H, H-C(1,8),
H-C(2,7)), 3.59 (m, 2 H, CH2(1′)), 2.82–2.78 (m, 2 H,
Following GP2 from 20 (3.4 g, 7.13 mmol) 30 (3.1 g,
97%) was obtained as a yellow, amorphous solid; UV-vis
(methanol): kmax (log e) = 283 nm (4.71); IR (KBr):
m = 2924m, 1633s, 1592s, 1569s, 1534m, 1469s, 1339w,
1271m, 1191m, 1170w, 857w, 758m cm–1; 1H NMR
3
(400 MHz, D2O): d = 7.68 (d, JH,H = 8.72 Hz, 2 H,
H-C(4,5)), 7.55 (t, 3JH,H = 7.68 Hz, 2 H, H-C(3,6)), 7.14 (d,
3JH,H = 8.51 Hz, 2 H, H-C(1,8)), 7.14 (m, 2 H, H-C(2,7)),

R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
983
3.52–3.48 (m, 2 H, CH2(1′)), 2.81–2.78 (m, 2 H, CH2(12′)),
1.53–1.43 (m, 4 H, 2 × CH2(2′,11′)), 1.14–0.98 (m, 16 H, 8 ×
CH2(3′,4′,5′,6′,7′,8′,9′,10′)); 13C NMR (100 MHz, D2O):
d = 156.0 (quart.), 135.1 (CH), 118.1 (CH), 48.5 (CH2(1′)),
39.7 (CH2(12′)), 29.1 (CH2), 29.0 (CH2), 28.9 (CH2), 28.9
(CH2), 28.5 (CH2), 28.4 (CH2), 27.0 (CH2), 26.1 (CH2), 25.9
(CH2); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z = 189
(39%) [MH2]2+, 378 (100%) [MH]+.
(log e) = 281 nm (4.70); IR (KBr): m = 3314m, 2981w, 1737s,
1669s, 1560w, 1522s, 1474w, 1430w, 1382w, 1342w,
1261m, 1198s, 1141m, 1028m, 759m, 651w, 1170w, 857w,
758m cm–1; 1H NMR (400 MHz, CDCl3): d = 8.85 (t,
3
3JH,H = 6.64 Hz, 1 H, NH), 8.23 (d, JH,H = 8.72 Hz, 2 H,
H-C(4,5)), 8.05 (d, 3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.62 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.31 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.08–4.01
(m, 8 H, 3 × CH2(Et), CH2(1′)), 3.77–3.72 (m, 2 H, CH2(2′)),
3.39 (s, 4 H, 2 × CH2(5″)), 3.33 (s, 2 H, CH2(4″)), 3.21 (s,
2 H, CH2(3″)), 2.67 (s, 4 H, 2 × CH2(1″,2″)), 1.22–1.16 (m,
9 H, 3 × CH3(Et)); 13C NMR (125 MHz, CDCl3): d = 173.0
(C=O), 170.7 (C=O), 170.6 (C=O), 152.4 (quart.), 147.5
(quart.), 130.0 (CH), 126.8 (CH), 123.5 (CH), 122.0 (CH),
115.0 (quart.), 60.3 (CH2(Et)), 60.2 (CH2(Et)), 57.9
(CH2(4″)), 56.0 (CH2(3″)), 55.0 (CH2(5″)), 53.6 (CH2(1″)),
52.2 (CH2(2″)), 50.1 (CH2(1′)), 39.7 (CH2(2′)), 13.7
(CH3(Et)); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol):
m/z = 596 (100%) [MH]+, 618 (7%) [MNa]+; HRMS
(C31H41N5O7): 595.30060; found: 595.30061; Anal.
C31H41N5O7 (C, H, N).
5.2.21. Ethylenediaminetetraacetic acid triethyl ester (31)
To a solution of EDTA (15.0 g, 51.3 mmol) in ethanol
(600 ml) conc. sulfuric acid (10 ml) was added and the
mixture was heated under reflux for 6 h. After cooling to
room temperature the reaction mixture was neutralized by
the careful addition of NaHCO3 and the solvents were remo-
ved under reduced pressure. The residue was suspended in
water (200 ml) and extracted with chloroform (2 × 150 ml).
The combined organic phases were dried (Na2SO4), the sol-
vent was evaporated and EDTA-tetraethyl ester 32 (14.5 g,
70%) was obtained [mp 30–32 °C (lit.: 34 °C [36], 32 °C
[37]); IR (KBr): m = 2982s, 2940m, 1736s, 1614w, 1447m,
1421m, 1368s, 1348s, 1190s, 1030s, 862w, 749m cm–1; 1H
NMR (500 MHz, CDCl3): d = 4.12 (q, 3J H,H = 7.10 Hz, 8 H,4
5.2.22. Ethyl 2-((2-{[3-(9-acridinylamino)propyl]amino}-
2-oxoethyl){2-[di-(2-ethoxy-2-oxoethyl)amino]ethyl}
amino) acetate (34)
× CH2(2)), 3.56 (s, 8 H,4 × CH2(3)), 2.87 (s, 4 H,2 × CH2(4)),
3
1.23 (t, J
= 7.10 Hz, 12 H, 4 × CH3(1)); 13C NMR
H,H
(125 MHz, CDCl3): d = 171.3 (CO), 60.4 (C-2), 55.2 (C-3),
52.3 (C-4), 14.2 (C-1); MS (ESI, 4.1 kV, 8 µl/min, N2,
methanol): m/z = 405 (19%) [MH]+, 427 (100%) [MNa]+].
To an emulsion of 32 (20 g, 49.0 mmol) in water (1500 ml)
containing Na2HPO4 (39.2 g, 276 mmol) and KH2PO4
(1.16 g, 8.5 mmol) at 27 °C PLE (BioChemica, 2.5 ml
(3500 units)) were added and the mixture was stirred for 8 h.
After extraction with hexane (2 × 50 ml, discarded) the aq.
layer was extracted with dichloromethane (5 × 100 ml) and
the combined organic phases were dried (MgSO4), and the
solvent was evaporated to yield 31 (14.5 g, 78%) as a viscous
oil [38]. IR (KBr): m = 2983m, 2938m, 1738s, 1634m, 1378s,
Following GP3 from 31 (930 mg, 2.47 mmol), isobutyl
chloroformate (337 mg, 2.47 mmol) and 22 (670 mg,
2.96 mmol) 34 (845 mg, 47%) was obtained as a syrup; RF
(methanol/ethyl acetate 1:6.4) 0.53; UV-vis (methanol): kmax
(log e) = 282 nm (4.72); IR (KBr): m = 3262m, 2981w, 1738s,
1635m, 1590m, 1538m, 1471m, 1372w, 1273w, 1196s,
1
1028m, 754w, 661w cm–1; H NMR (400 MHz, CDCl3):
d = 8.96–8.91 (m, 1 H, NH), 8.49 (d, 3JH,H = 8.72 Hz, 2 H,
H-C(4,5)), 8.28 (d, 3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.72 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.37 (dd,
3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.18–4.10
(m, 6 H, 3 × CH2(Et)), 4.07–4.04 (m, 2 H, CH2(1′)), 3.63–
3.56 (m, 2 H, CH2(3′)), 3.52 (s, 4 H, 2 × CH2(5″)), 3.45 (s,
2 H, CH2(4″)), 3.43 (s, 2 H, CH2(3″)), 2.81 (s, 4 H, 2 ×
CH2(1″,2″)), 2.16–2.11 (m, 2 H, CH2(2′)), 1.27–1.22 (m, 9 H,
3 × CH3(Et)); 13C NMR (125 MHz, CDCl3): d =173.3
(C=O), 171.1 (C=O), 171.0 (C=O), 156.9 (quart.), 140.6
(quart.), 133.9 (CH), 123.8 (CH), 123.6 (CH), 121.0 (CH),
60.8 (CH2(Et)), 60.7 (CH2(Et)), 58.3 (CH2(4″)), 55.8
(CH2(3″)), 54.7 (CH2(5″)), 53.0 (CH2(1″)), 52.3 (CH2(2″)),
45.0 (CH2(1′)), 36.1 (CH2(3′)), 30.6 (CH2(2′)), 14.3
(CH3(Et)); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol):
m/z = 610 (100%) [MH]+, 632 (2%) [MNa]+; HRMS
C32H43N5O7: 609.31625; found: 609.31627; Anal.
C32H43N5O7 (C, H, N).
1
1199s, 1097s, 1028m, 865w, 734w, 588w cm–1; H NMR
(500 MHz, CDCl3): d = 4.15 (m, 3JH,H = 7.10 Hz, 6 H, 3 ×
CH2), 3.53 (s, 4 H, 2 × CH2), 3.47 (s, 2 H, CH2), 3.45 (s, 2 H,
CH2), 2.84 (s, 4 H, CH2), 1.24 (t, 3JH,H = 7.10 Hz, 9 H, 3 ×
CH3); 13C NMR (125 MHz, CDCl3): d = 173.0 (CO), 170.9
(CO), 170.6 (CO), 70.0 (C-7), 60.8 (C-6), 57.0 (C-5), 56.1
(C-4), 54.6 (C-3), 52.5 (C-1), 51.7 (C-2), 14.1 (C-8,9); MS
(ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z = 377 (39%)
[MH]+, 399 (100%) [MNa]+, 415 (28%) [MK]+, 791 (93%)
[M2K]+; HRMS (C16H29N2O8): 376.14857; found:
376.14859.
5.2.21. Ethyl 2-((2-{[2-(9-acridinylamino)ethyl]amino}-2-
oxoethyl){2-[di-(2-ethoxy-2-oxoethyl)amino]ethyl} acetate
(33)
Following GP3 from 31 (1.2 g, 2.67 mmol), isobutyl
chloroformate (438 mg, 3.2 mmol) and 21 (830 mg,
2.67 mmol) 33 (600 mg, 36%) was obtained as a syrup; RF
(methanol/ethyl acetate 1:6.4) 0.63; UV-vis (methanol): kmax
5.2.23. Ethyl 2-((2-{[4-(9-acridinylamino)butyl]amino}-2-
oxoethyl){2-[di-(2-ethoxy-2-oxoethyl)amino]ethyl} amino)
acetate (35)
Following GP3 from 23 (900 mg, 2.66 mmol), 31 (1.2 g,
3.19 mmol) and isobutyl chloroformate (435 mg, 3.19 mmol)

984
R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
35 (720 mg, 43%) was obtained as a viscous oil; RF
(methanol/ethyl acetate 1:6.4) 0.50; UV-vis (methanol): kmax
(log e) = 283 nm (4.65); IR (KBr): m = 3280s, 2981w, 2920w,
1736s, 1657m, 1562m, 1473m, 1272w, 1195s, 1027m, 758w,
661w cm–1; 1H NMR (500 MHz, CDCl3): d = 8.16 (t,
5.2.28. Ethyl 2-((2-{[6-(9-acridinylamino)hexyl]amino}-2-
oxoethyl){2-[di-(2-ethoxy-2-oxoethyl)amino]ethyl} amino)
acetate (37)
Following GP3 from 25 (811 mg, 2.06 mmol), 31
(930 mg, 2.47 mmol) and isobutyl chloroformate (337 mg,
2.47 mmol) 37 (600 mg, 45%) was obtained as a viscous oil.
RF (methanol/ethyl acetate 1:6.4) 0.47; UV-vis (methanol):
kmax(log e) = 281 nm (4.55); IR (KBr): m = 3325m, 2981w,
2934w, 2857w, 1738s, 1667s, 1561m, 1522m, 1428m,
1383w, 1344m, 1261m, 1199s, 1028m, 860w, 762w, 652w
cm–1; 1H NMR (500 MHz, CDCl3): d = 8.07 (d,
3
3JH,H = 6.42 Hz, 1 H, NH), 8.11 (d, JH,H = 8.71 Hz, 2 H,
H-C(4,5)), 7.98 (d, 3JH,H = 8.71 Hz, 2 H, H-C(1,8)), 7.57 (dd,
3
3JH,H = 8.71 Hz, JH,H = 7.33 Hz, 2 H, H-C(3,6)), 7.28 (t,
3JH,H = 7.56 Hz, 2 H, H-C(2,7)), 4.11–4.04 (m, 6 H, 3 ×
CH2(Et)), 3.86–3.83 (m, 2 H, CH2(1′)), 3.47 (s, 4 H, 2 ×
CH2(5″)), 3.36 (s, 2 H, CH2(4″)), 3.34–3.30 (m, 2 H,
CH2(4′)), 3.26 (s, 2 H, CH2(3″)), 2.79–2.72 (m, 4 H, 2 ×
CH2(1″,2″)), 1.86–1.81 (m, 2 H, CH2(2′)), 1.71–1.67 (m, 2 H,
CH2(3′)), 1.21–1.16 (m, 9 H, 3 × CH3(Et)); 13C NMR
(125 MHz, CDCl3): d = 171.6 (C=O), 171.3 (C=O), 171.0
(C=O), 152.1 (quart.), 147.9 (quart.), 130.2 (CH), 128.0
(CH), 123.2 (CH), 122.8 (CH), 116.0 (quart.), 60.7
(CH2(Et)), 60.6 (CH2(Et)), 58.7 (CH2(4″)), 56.0 (CH2(3″)),
54.7 (CH2(5″)), 53.0 (CH2(1″)), 52.2 (CH2(2″)), 50.2
(CH2(1′)), 38.5 (CH2(4′)), 28.6 (CH2), 27.1 (CH2), 14.1
(CH3(Et)); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol):
m/z = 624 (100%) [MH]+, 646 (7%) [MNa]+; HRMS
(C33H45N5O7): 623.33190: found: 623.33193; Anal.
C33H45N5O7 (C, H, N).
3
3JH,H = 8.71 Hz, 2 H, H-C(4,5)), 7.97 (d, JH,H = 8.71 Hz,
2 H, H-C(1,8)), 7.58 (dd, 3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz,
2 H, H-C(3,6)), 7.28 (dd, 3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz,
2 H, H-C(2,7)), 4.12–4.06 (m, 6 H, 3 × CH2(Et)), 3.79–3.76
(m, 2 H, CH2(1′)), 3.46 (s, 4 H, 2 × CH2(5″)), 3.35 (s, 2 H,
CH2(4″)), 3.23 (s, 2 H, CH2(3″)), 3.22–3.17 (m, 2 H,
CH2(6′)), 2.74 (m, 4 H, 2 × CH2(1″,2″)), 1.79–1.73 (m, 2 H,
CH2(2′)), 1.53–1.41 (m, 4 H, 2 × CH2(3′,5′)), 1.37–1.31 (m,
2 H, CH2(4′)), 1.24–1.1.7 (m, 9 H, 3 × CH3(Et)); 13C NMR
(125 MHz, CDCl3): d = 171.3 (C=O), 171.1 (C=O), 170.9
(C=O), 151.8 (quart.), 149.0 (quart.), 130.1 (CH), 128.5
(CH), 122.9 (CH), 122.8 (CH), 116.1 (quart.), 60.6
(CH2(Et)), 60.5 (CH2(Et)), 58.7 (CH2(4″)), 55.9 (CH2(3″)),
54.8 (CH2(5″)), 52.9 (CH2(1″)), 52.2 (CH2(2″)), 50.4
(CH2(1′)), 38.7 (CH2(6′)), 31.4 (CH2), 29.4 (CH2), 26.4
(CH2), 26.3 (CH2), 14.1 (CH3(Et)); MS (ESI, 4.1 kV,
8 µl/min, N2, methanol): m/z = 652 (100%) [MH]+; HRMS
(C35H49N5O7): 651.36320; found: 651.36322; Anal.
C35H49N5O7 (C, H, N).
5.2.24. Ethyl 2-((2-{[5-(9-acridinylamino)pentyl]amino}-2-
oxoethyl){2-[di-(2-ethoxy-2-oxoethyl)amino]ethyl} amino)
acetate (36)
Following GP3 from 24 (1.0 g, 2.84 mmol), 31 (1.26 g,
3.36 mmol) and isobutyl chloroformate (458 mg, 3.36 mmol)
36 (950 mg, 52%) was obtained as a viscous oil; RF
(methanol/ethyl acetate 1:6.4) 0.46; UV-vis (methanol): kmax
(log e) = 283 nm (4.73); IR (KBr): m = 3324m, 2981w,
2935w, 2860w, 1738s, 1660s, 1616w, 1560m, 1522s, 1474m,
1372w, 1260m, 1196s, 1028m, 859w, 761m, 651w cm–1; 1H
NMR (500 MHz, CDCl3): d = 8.01 (d, 3JH,H = 8.71 Hz, 2 H,
H-C(4,5)), 8.06–8.03 (m, 1 H, NH), 8.02 (d, 3JH,H = 8.71 Hz,
2 H, H-C(1,8)), 7.62 (dd, 3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz,
2 H, H-C(3,6)), 7.32 (dd, 3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz,
2 H, H-C(2,7)), 4.10–4.04 (m, 6 H, 3 × CH2(Et)), 3.81–3.78
(m, 2 H, CH2(1′)), 3.46 (s, 4 H, 2 × CH2(5″)), 3.35 (s, 2 H,
CH2(4″)), 3.26 (s, 2 H, CH2(3″)), 3.28–3.24 (m, 2 H,
CH2(5′)), 2.78–2.71 (m, 4 H, 2 × CH2(1″,2″)), 1.84–1.78 (m,
2 H, CH2(2′)), 1.58–1.53 (m, 2 H, CH2(4′)), 1.49–1.45 (m,
2 H, CH2(3′)), 1.20–1.16 (m, 9 H, 3 × CH3(Et)); 13C NMR
(100 MHz, CDCl3): d = 171.2 (C=O), 171.1 (C=O), 170.8
(C=O), 151.2 (quart.), 149.2 (quart.), 129.6 (CH), 129.5
(CH), 122.8 (CH), 122.7 (CH), 116.5 (quart.), 60.7
(CH2(Et)), 60.6 (CH2(Et)), 58.7 (CH2(4″)), 56.0 (CH2(3)),
54.8 (CH2(5″)), 53.0 (CH2(1″)), 52.3 (CH2(2″)), 50.9
(CH2(1′)), 38.7 (CH2(5′)), 31.2 (CH2), 29.5 (CH2), 24.2
(CH2), 14.3 (CH3(Et)); MS (ESI, 4.1 kV, 8 µl/min, N2,
methanol): m/z = 638 (100%) [MH]+, 660 (5%) [MNa]+;
HRMS (C34 H47N5O7): 637.34755; found: 637.34758; Anal.
5.2.25. Ethyl 2-((2-{[7-(9-acridinylamino)heptyl]amino}-2-
oxoethyl){2-[di-(2-ethoxy-2-oxoethyl)amino]ethyl} amino)
acetate (38)
Following GP3 from 26 (665 mg, 1.75 mmol), 31
(788 mg, 2.09 mmol) and isobutyl chloroformate (287 mg,
2.09 mmol) 38 (490 mg, 42%) was obtained as a viscous oil.
RF (methanol/ethyl acetate 1:6.4) 0.53; UV-vis (methanol):
kmax (log e) = 283 nm (4.66); IR (KBr): m = 3335s, 2932w,
2857w, 1738s, 1667s, 1560s, 1521m, 1473m, 1383m,
1344m, 1260s, 1197s, 1028m, 858m, 761w, 651w cm–1; 1H
NMR (400 MHz, CDCl3): d = 8.06 (d, 3JH,H = 8.72 Hz, 2 H,
H-C(4,5)), 8.00 (d, 3JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.90 (t,
3
3JH,H = 5.81 Hz, 1 H, NH), 7.61 (dd, JH,H = 8.72 Hz,
3JH,H = 7.47 Hz, 2 H, H-C(3,6)), 7.30 (dd, 3JH,H = 8.72 Hz,
3JH,H = 7.47 Hz, 2 H, H-C(2,7)), 4.12–4.04 (m, 6 H, 3 ×
CH2(Et)), 3.77–3.74 (m, 2 H, CH2(1′)), 3.46 (s, 4 H, 2 ×
CH2(5″)), 3.35 (s, 2 H, CH2(4″)), 3.23 (s, 2 H, CH2(3″)),
3.22–3.16 (m, 2 H, CH2(7′)), 2.79–2.70 (m, 4 H, 2 ×
CH2(1″,2″)), 1.77–1.71 (m, 2 H, CH2(2′)), 1.48–1.43 (m, 2 H,
CH2(6′)), 1.42–1.33 (m, 2 H, CH2(3′)), 1.30–1.28 (m, 4 H,
2 × CH2(4′,5′)), 1.21–1.16 (m, 9 H, 3 × CH3(Et)); 13C NMR
(100 MHz, CDCl3): d = 171.0 (C=O), 170.9 (C=O), 170.7
(C=O), 151.3 (quart.), 148.9 (quart.), 129.7 (CH), 129.0
(CH), 122.8 (CH), 122.7 (CH), 116.4 (quart.), 60.6
(CH2(Et)), 60.5 (CH2(Et)), 58.7 (CH2(4″)), 55.9 (CH2(3″)),
C
34 H47N5O7 (C, H, N).

R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
985
54.8 (CH2(5″)), 52.9 (CH2(1″)), 52.2 (CH2(2″)), 50.8
(CH2(1′)), 39.0 (CH2(7′)), 31.6 (CH2), 29.5 (CH2), 28.9
(CH2), 26.8 (CH2), 26.7 (CH2), 14.2 (CH3(Et)); MS (ESI,
4.1 kV, 8 µl/min, N2, methanol): m/z = 666 (100%) [MH]+,
688 (5%) [MNa]+; HRMS (C36H51N5O7): 665.37885; found:
665.37887; Anal. C36H51N5O7 (C, H, N).
3.25 (s, 2 H, CH2(3″)), 3.23–3.19 (m, 2 H, CH2(9′)), 2.81–
2.73 (m, 4 H, 2 × CH2(1″,2″)), 1.79–1.73 (m, 2 H, CH2(2′)),
1.48–1.45 (m, 2 H, CH2(8′)), 1.44–1.39 (m, 2 H, CH2(3′)),
1.33–1.26 (m,
8 H, 4 × CH2(4′,5′,6′,7′), 1.22 (t,
3JH,H = 7.33 Hz, 9 H, 3 × CH3(Et)); 13C NMR (125 MHz,
CDCl3): d = 171.3 (C=O), 171.2 (C=O), 171.0 (C=O), 151.6
(quart.), 149.0 (quart.), 129.9 (CH), 129.1 (CH), 122.9 (CH),
122.7 (CH), 116.4 (quart.), 60.7 (CH2(Et)), 60.5 (CH2(Et)),
58.7 (CH2(4″)), 56.0 (CH2(3″)), 54.9 (CH2(5″)), 53.0
(CH2(1″)), 52.3 (CH2(2″)), 50.8 (CH2(1′)), 39.1 (CH2(9′)),
31.7 (CH2), 29.6 (CH2), 29.3 (CH2), 29.2 (CH2), 29.1 (CH2),
26.83 (CH2), 26.78 (CH2), 14.2 (CH3(Et)); MS (ESI, 4.1 kV,
8 µl/min, N2, methanol): m/z = 347 (9%) [MH2]2+, 694
(100%) [MH]+, 716 (7%) [MNa]+; HRMS (C38H55N5O7):
693.41015; found: 693.41016;Anal. C38 H55N5O7 (C, H, N).
5.2.26. Ethyl 2-((2-{[8-(9-acridinylamino)octyl]amino}-2-
oxoethyl){2-[di-(2-ethoxy-2-oxoethyl)amino]ethyl} amino)
acetate (39)
Following GP3 from 27 (1.2 g, 3.04 mmol), 31 (1.35 g,
3.6 mmol) and isobutyl chloroformate (492 mg, 3.6 mmol)
39 (865 mg, 42%) was obtained as a viscous oil. RF
(methanol/ethyl acetate 1:6.4) 0.60; UV-vis (methanol): kmax
(log e) = 281 nm (4.73); IR (KBr): m = 3328s, 2930m, 2855m,
1739s, 1661s, 1560s, 1521m, 1474m, 1370m, 1344m, 1260s,
1195s, 1028s, 858m, 761m, 652w cm–1; 1H NMR (500 MHz,
CDCl3): d = 8.01 (d, 3JH,H = 8.71 Hz, 2 H, H-C(4,5)), 7.95 (d,
3JH,H = 8.71 Hz, 2 H, H-C(1,8)), 7.87 (m, 1 H, NH), 7.54 (dd,
3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz, 2 H, H-C(3,6)), 7.23 (dd,
3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz, 2 H, H-C(2,7)), 4.07–4.02
(m, 6 H, 3 × CH2(Et)), 3.70–3.67 (m, 2 H, CH2(1′)), 3.42 (s,
4 H, 2 × CH2(5″)), 3.31 (s, 2 H, CH2(4″)), 3.19 (s, 2 H,
CH2(3″)), 3.16–3.12 (m, 2 H, CH2(8′)), 2.74–2.66 (m, 4 H,
2 × CH2(1″,2″)), 1.68–1.62 (m, 2 H, CH2(2′)), 1.41–1.39 (m,
2 H, CH2(7′)), 1.31–1.28 (m, 2 H, CH2(3′)), 1.21–1.18 (m,
5.2.28. Ethyl 2-((2-{[10-(9-acridinylamino)decyl]amino}-
2-oxoethyl){2-[di-(2-ethoxy-2-oxoethyl)amino]ethyl}
amino) acetate (41)
Following GP3 from 29 (900 mg, 2.13 mmol), 31
(961 mg, 2.56 mmol) and isobutyl chloroformate (350 mg,
2.56 mmol) 41 (605 mg, 40%) was obtained as a viscous oil.
RF (methanol/ethyl acetate 1:6.4) 0.64; UV-vis (methanol):
kmax (log e) = 283 nm (4.69); IR (KBr): m = 2928m, 2854m,
1740s, 1670s, 1560w, 1521m, 1474m, 1370m, 1260s,
1195m, 1029m, 858m, 760w, 651w cm–1; 1H NMR
6 H, 3 × CH2(4′,5′,6′)), 1.16–1.12 (m, 9 H, 3 × CH3(Et)); 13
C
(400 MHz, CDCl3): d = 8.01 (d, JH,H = 8.72 Hz, 2 H,
3
3
NMR (125 MHz, CDCl3): d = 171.1 (C=O), 171.0 (C=O),
170.8 (C=O), 151.3 (quart.), 148.9 (quart.), 129.6 (CH),
128.8 (CH), 122.7 (CH), 122.6 (CH), 116.3 (quart.), 60.5
(CH2(Et)), 60.3 (CH2(Et)), 58.5 (CH2(4″)), 55.7 (CH2(3″)),
54.7 (CH2(5″)), 52.7 (CH2(1″)), 52.0 (CH2(2″)), 50.6
(CH2(1′)), 38.8 (CH2(8′)), 31.4 (CH2), 29.4 (CH2), 28.9
(CH2), 28.8 (CH2), 26.5 (CH2), 14.0 (CH3(Et)); MS (ESI,
4.1 kV, 8 µl/min, N2, methanol): m/z = 340 (8%) [MH2]2+,
680 (100%) [MH]+, 703 (9%) [MNa]+; HRMS
(C37H53N5O7): 679.39450; found: 679.39452; Anal.
C37H53N5O7 (C, H, N).
H-C(4,5)), 7.90 (d, JH,H = 8.72 Hz, 2 H, H-C(1,8)), 7.87–
7.82 (m, 1 H, NH), 7.50 (dd, 3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz,
2 H, H-C(3,6)), 7.20 (dd, 3JH,H = 8.72 Hz, 3JH,H = 7.47 Hz,
2 H, H-C(2,7)), 4.04 (q, 3JH,H = 7.05 Hz, 6 H, 3 × CH2(Et)),
3.71–3.68 (m, 2 H, CH2(1′)), 3.42 (s, 4 H, 2 × CH2(5″)), 3.31
(s, 2 H, CH2(4″)), 3.18 (s, 2 H, CH2(3″)), 3.17–3.12 (m, 2 H,
CH2(10′)), 2.72–2.65 (m, 4 H, 2 × CH2(1″,2″)), 1.70–1.63
(m, 2 H, CH2(2′)), 1.42–1.37 (m, 2 H, CH2(9′)), 1.34–1.27
(m,
2
H, CH2(3′)), 1.20–1.12 (m, 10 H,
5
×
3
CH2(4′,5′,6′,7′,8′), 1.14 (t, JH,H = 7.05 Hz, 9 H,2 ×
CH3(Et)); 13C NMR (100 MHz, CDCl3): d = 170.9 (C=O),
170.7 (C=O), 170.6 (C=O), 151.5 (quart.), 148.0 (quart.),
129.7 (CH), 128.0 (CH), 122.8 (CH), 122.4 (CH), 115.9
(quart.), 60.4 (CH2(Et)), 60.3 (CH2(Et)), 58.5 (CH2(4″)),
55.8 (CH2(3″)), 54.7 (CH2(5″)), 52.8 (CH2(1″)), 52.1
(CH2(2″)), 50.4 (CH2(1′)), 39.0 (CH2(10′)), 31.4 (CH2), 29.5
(CH2), 29.2 (CH2), 29.1 (CH2), 29.08 (CH2), 26.06 (CH2),
26.7 (CH2), 14.1 (CH3(Et)); MS (ESI, 4.1 kV, 8 µl/min, N2,
methanol): m/z = 708 (100%) [MH]+, 730 (7%) [MNa]+;
HRMS (C39H57N5O7): 707.42580; found: 707.42583; Anal.
C39H57N5O7 (C, H, N).
5.2.27. Ethyl 2-((2-{[9-(9-acridinylamino)nonyl]amino}-2-
oxoethyl){2-[di-(2-ethoxy-2-oxoethyl)amino]ethyl} amino)
acetate (40)
Following GP3 from 28 (1.2 g, 2.94 mmol), 31 (1.36 g,
3.52 mmol) and isobutyl chloroformate (482 mg, 3.52 mmol)
40 (790 mg, 39%) was obtained as a viscous oil. RF
(methanol/ethyl acetate 1:6.4) 0.63; UV-vis (methanol): kmax
(log e) = 281 nm (4.69); IR (KBr): m = 3323m, 2928m,
2854m, 1738s, 1667s, 1561m, 1521m, 1384m, 1261s,
1
1196m, 1027m, 759m cm–1; H NMR (500 MHz, CDCl3):
3
d = 8.08 (d, JH,H = 8.71 Hz, 2 H, H-C(4,5)), 8.03 (d,
5.2.29. Ethyl 2-((2-{[12-(9-acridinylamino)dodecyl]
amino}-2-oxoethyl){2-[di-(2-ethoxy-2-oxoethyl)amino]
ethyl}amino) acetate (42)
Following GP3 from 30 (930 mg, 2.06 mmol), 31
(930 mg, 2.47 mmol) and isobutyl chloroformate (337 mg,
2.47 mmol) 42 (730 mg, 48%) was obtained as a viscous oil.
RF (methanol/ethyl acetate 1:6.4) 0.66; UV-vis (methanol):
3JH,H = 8.71 Hz, 2 H, H-C(1,8)), 7.91–7.87 (m, 1 H, NH),
7.62 (dd, 3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz, 2 H, H-C(3,6)),
7.32 (dd, 3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz, 2 H, H-C(2,7)),
3
4.12 (q, JH,H = 7.33 Hz, 2 H, CH2(Et)), 4.11 (q,
3JH,H = 7.33 Hz, 4 H, 2 × CH2(Et)), 3.80–3.77 (m, 2 H,
CH2(1′)), 3.49 (s, 4 H, 2 × CH2(5″)), 3.38 (s, 2 H, CH2(4″)),

986
R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
kmax (log e) = 281 nm (4.74); IR (KBr): m = 2927m, 2854w,
1739s, 1667s, 1561m, 1522w, 1474w, 1373w, 1260s, 1196m,
1029m, 858m, 761w, 651w cm–1; 1H NMR (500 MHz,
CDCl3): d = 8.08 (d, 3JH,H = 8.74 Hz, 2 H, H-C(4,5)), 8.04 (d,
3JH,H = 8.74 Hz, 2 H, H-C(1,8)), 7.90–7.85 (m, 1 H, NH),
7.64 (dd, 3JH,H = 8.74 Hz, 3JH,H = 7.49 Hz, 2 H, H-C(3,6)),
7.34 (dd, 3JH,H = 8.74 Hz, 3JH,H = 7.49 Hz, 2 H, H-C(2,7)),
4.16–4.10 (m, 2 H, CH2(Et)), 3.81–3.78 (m, 2 H, CH2(1′)),
3.50 (s, 4 H, CH2(5″)), 3.39 (s, 2 H, CH2(4″)), 3.26 (s, 2 H,
CH2(3″)), 3.24–3.19 (m, 2 H, CH2(12′)), 2.82–2.74 (m, 4 H,
2 × CH2(1″,2″)), 1.78–1.74 (m, 2 H, CH2(2′)), 1.50–1.45 (m,
2 H, CH2(11′)), 1.44–1.39 (m, 2 H, CH2(3′)), 1.32–1.22 (m,
14 H, 7 × CH2(4′,5′,6′,7′,8′,9′,10′), 1.25–1.21 (m, 9 H, 3 ×
CH3(Et)); 13C NMR (125.7 MHz, CDCl3): d = 171.3 (C=O),
171.1 (C=O), 171.0 (C=O), 151.4 (quart.), 129.8 (CH), 123.0
(CH), 122.7 (CH), 116.6 (quart.), 60.7 (CH2(Et)), 60.6
(CH2(Et)), 58.8 (CH2(4″)), 56.0 (CH2(3″)), 54.9 (CH2(5″)),
53.0 (CH2(1″)), 52.3 (CH2(2″)), 51.0 (CH2(1′)), 39.2
(CH2(12′)), 31.8 (CH2), 29.7 (CH2), 29.52 (CH2), 29.46
(CH2), 29.45 (CH2), 29.3 (CH2), 27.0 (CH2), 26.9 (CH2),
14.2 (CH3(Et)); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol):
m/z = 368 (2%) [MH2]2+, 736 (100%) [MH]+, 758 (6%)
[MNa]+; HRMS (C41H61N5O7): 735.45710; found:
735.45711; Anal. C41H61N5O7 (C, H, N).
3JH,H = 8.71 Hz, 2 H, H-C(4,5)), 7.82–7.80 (m, 4 H,
H-C(1,8), H-C(3,6)), 7.48–7.43 (m, 2 H, H-C(2,7)), 4.11–
4.07 (m, 2 H, CH2(1′)), 3.39–3.36 (m, 2 H, CH2(3′)), 3.19 (s,
4 H, 2 × CH2(5″)), 3.12 (s, 2 H, CH2(4″)), 3.05 (s, 2 H,
CH2(3″)), 2.70–2.52 (m, 4 H, 2 × CH2(1″,2″)), 2.15–2.08 (m,
2 H, CH2(2′)); 13C NMR (125 MHz, D2O): d = 179.0 (C=O),
174.1 (C=O), 154.6 (quart.), 140.7 (quart.), 133.6 (CH),
123.7 (CH), 123.3 (CH), 120.2 (CH), 111.8 (quart.), 58.8
(CH2(3″)), 58.4 (CH2(5″)), 58.2 (CH2(4″)), 52.1 (CH2(1″)),
51.9 (CH2(2″)), 45.9 (CH2(1′)), 36.2 (CH2(3′)), 28.9
(CH2(2′)); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol):
m/z = 524 (100%) [M – H]–; HRMS (C26H31N5O7):
525.22235; found: 525.22237.
5.2.32. 2-[{2-[(2-{[4-(9-Acridinylamino)butyl]amino}-2-
oxoethyl)(carboxymethyl)-amino]ethyl}(carboxy-
methyl)amino]acetic acid (45)
Following GP4 from 35 (136 mg, 022 mmol) 45 (100 mg,
87%) was obtained as an orange-coloured, amorphous solid.
UV-vis (methanol): kmax (log e) = 284 nm (4.60); IR (KBr):
m = 3424s, 2927m, 2853m, 1636s, 1589s, 1471m, 1404m,
1336m, 1272m, 1191w, 1170w, 1121w cm–1; 1H NMR
3
(500 MHz, D2O): d = 7.52 (d, JH,H = 8.71 Hz, 2 H,
H-C(4,5)), 7.43 (dd, 3JH,H = 8.71 Hz, 3JH,H = 6.88 Hz, 2 H,
H-C(3,6)), 7.04 (d, 3JH,H = 8.71 Hz, 2 H, CH(1,8)), 7.02 (dd,
3JH,H = 8.71 Hz, 3JH,H = 6.88 Hz, 2 H, H-C(2,7)), 3.36–3.33
(m, 2 H, CH2(1′)), 3.17 (s, 4 H, 2 × CH2(5″)), 3.11–3.07 (m,
2 H, CH2(4′)), 3.08 (s, 2 H, CH2(4″)), 3.01 (s, 2 H, CH2(3″)),
2.69–2.63 (m, 2 H, CH2(1″)), 2.60–2.55 (m, 2 H, CH2(2″)),
1.50–1.46 (m, 2 H, CH2(2′)), 1.43–1.39 (m, 2 H, CH2(3′));
13C NMR (125 MHz, D2O): d = 178.9 (C=O), 173.9 (C=O),
154.6 (quart.), 140.3 (quart.), 133.8 (CH), 123.9 (CH), 123.3
(CH), 119.7 (CH), 111.7 (quart.), 58.7 (CH2(3″)), 58.3
(CH2(5″)), 58.1 (CH2(4″)), 52.0 (CH2(1″)), 51.7 (CH2(2″)),
48.0 (CH2(1′)), 38.4 (CH2(4′)), 25.5 (CH2), 25.7 (CH2); MS
(ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z = 538 (100%) [M
– H]–; HRMS (C27H33N5O7): 539.58022; found: 539.58023.
5.2.30. 2-[{2-[(2-{[2-(9-Acridinylamino)ethyl]amino}-2-
oxoethyl)(carboxymethyl)-amino]ethyl}(carboxymethyl)
amino] acetic acid (43)
Following GP4 from 33 (180 mg, 0.302 mmol) 43
(129 mg, 84%) was obtained as an amorphous orange-red
solid. UV-vis (methanol): kmax (log e) = 282 nm (4.66); IR
(KBr): m = 3854s, 2832m, 1590s, 1474w, 1407s, 1337m,
1
1261w, 1172w, 1123w, 1024w cm–1; H NMR (500 MHz,
D2O): d = 7.53 (d, 3JH,H = 8.68 Hz, 2 H, H-C(4,5)), 7.42 (dd,
3
3JH,H = 8.68 Hz, JH,H = 7.37 Hz, 2 H, H-C(3,6)), 7.11 (d,
3JH,H = 8.55 Hz, 2 H, H-C(1,8)), 7.04 (dd, 3JH,H = 8.68 Hz,
3JH,H = 7.37 Hz, 2 H, H-C(2,7)), 3.59–3.54 (m, 2 H, CH2(1′)),
3.33–3.28 (m, 2 H, CH2(2′)), 3.01 (s, 4 H, 2 × CH2(5″)), 2.91
(s, 2 H, CH2(4″)), 2.83 (s, 2 H, CH2(3″)), 2.46–2.41 (m, 2 H,
CH2(1″)), 2.37–2.31 (m, 2 H, CH2(2″)); 13C NMR
(125 MHz, D2O): d = 178.8 (C=O), 175.1 (C=O), 154.1
(quart.), 142.1 (quart.), 133.1 (CH), 123.6 (CH), 123.3 (CH),
121.5 (CH), 112.6 (quart.), 58.5 (CH2(3″)), 58.3 (CH2(5″)),
57.8 (CH2(4″)), 51.9 (CH2(1″)), 51.7 (CH2(2″)), 48.7
(CH2(1′)), 39.1 (CH2(2′)); MS (ESI, 4.1 kV, 8 µl/min, N2,
methanol): m/z = 510 (100%) [M – H]–; HRMS
(C25H29N5O7): 511.20670; found: 511.20671.
5.2.33. 2-[{2-[(2-{[5-(9-Acridinylamino)pentyl]amino}-2-
oxoethyl)(carboxymethyl)- amino]ethyl}(carboxy-
methyl)amino] acetic acid (46)
Following GP4 from 36 (440 mg, 0.69 mmol) 46 (320 mg,
84%) was obtained as an orange-coloured, amorphous solid.
UV-vis (methanol): kmax (log e) = 284 nm (4.53); IR (KBr):
m = 3425s, 2935m, 1591s, 1473w, 1407m, 1337w, 1261w,
1
1171w, 1123w cm–1; H NMR (500 MHz, D2O): d = 7.28
(dd, 3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz, 2 H, H-C(3,6)), 7.24
3
(d, JH,H
= 8.71 Hz, 2 H, H-C(4,5)), 6.87 (dd,
3
5.2.31. 2-[{2-[(2-{[3-(9-Acridinylamino)propyl]amino}-2-
oxoethyl)(carboxymethyl)-amino]ethyl}(carboxymethyl)
amino]acetic acid (44)
3JH,H = 8.71 Hz, JH,H = 7.33 Hz, 2 H, H-C(2,7)), 6.76 (d,
3JH,H = 8.71 Hz, 2 H, H-C(1,8)), 3.33–3.26 (m, 4 H, 2 ×
CH2(1′,3″)), 3.13 (s, 2 H, CH2(4″)), 3.10–3.02 (m, 2 H,
CH2(5′)), 3.05 (s, 4 H, CH2(5″)), 2.81–2.72 (m, 2 H,
CH2(1″)), 2.69–2.63 (m, 2 H, CH2(2″)), 1.39–1.29 (m, 4 H,
2 × CH2(2′,4′)), 1.14–1.06 (m, 2 H, CH2(3′)); 13C NMR
(125 MHz, D2O): d = 178.8 (C=O), 173.4 (C=O), 154.4
(quart.), 138.6 (quart.), 134.4 (CH), 123.7 (CH), 123.4 (CH),
Following GP4 from 34 (740 mg, 1.2 mmol) 44 (548 mg,
86%) was obtained as an amorphous, orange-coloured solid.
UV-vis (methanol): kmax (log e) = 283 nm (4.63); IR (KBr):
m = 3423s, 1591s, 1474m, 1409s, 1337m, 1261m, 1171w,
1
1123m cm–1; H NMR (400 MHz, CD3OD): d = 8.46 (d,

R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
987
118.3 (CH), 110.7 (quart.), 58.5 (CH2(3″)), 58.3 (CH2(5″)),
5.2.36. 2-[{2-[(2-{[8-(9-Acridinylamino)octyl]amino}-2-
oxoethyl)(carboxymethyl)- amino]ethyl}(carboxy-
methyl)amino] acetic acid (49)
57.9 (CH2(4″)), 52.2 (CH2(1″)), 51.3 (CH2(2″)), 48.0
(CH2(1′)), 38.8 (CH2(5′)), 28.6 (CH2), 27.9 (CH2), 23.2
(CH2); MS (ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z = 552
(100%) [M – H]–; HRMS (C28H35N5O7): 553.25365; found:
553.25367.
Following GP4 from 39 (134 mg, 0.197 mmol) 49 (99 mg,
85%) was obtained as an amorphous, orange-coloured solid.
UV-vis (methanol): kmax (log e) = 284 nm (4.57); IR (KBr):
m = 3425s, 2853m, 1589s, 1473m, 1407m, 1336w, 1260w,
1170w, 1122w cm–1; 1H NMR (500 MHz, D2O): d = 7.57 (d,
3JH,H = 8.71 Hz, 2 H, H-C(4,5)), 7.45 (dd, 3JH,H = 8.71 Hz,
5.2.34. 2-[{2-[(2-{[6-(9-Acridinylamino)hexyl]amino}-2-
oxoethyl)(carboxymethyl)- amino]ethyl}(carboxy-
methyl)amino] acetic acid (47)
3
3JH,H = 7.33 Hz, 2 H, H-C(3,6)), 7.20 (d, JH,H = 8.71 Hz,
2 H, H-C(1,8)), 7.03 (dd, 3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz,
2 H, H-C(2,7)), 3.32–3.29 (m, 2 H, CH2(1′)), 3.14 (s, 4 H, 2 ×
CH2(5″)), 3.09 (s, 2 H, CH2(4″)), 3.03 (s, 2 H, CH2(3″)),
3.02–2.98 (m, 2 H, CH2(8′)), 2.66–2.55 (m, 4 H, 2 ×
CH2(1″,2″)), 1.43–1.37 (m, 2 H, CH2(2′)), 1.30–1.23 (m, 2 H,
CH2(7′)), 1.07–0.98 (m, 8 H, 4 × CH2(3′,4′,5′,6′)); 13C NMR
(125 MHz, D2O): d = 179.0 (C=O), 173.7 (C=O), 154.4
(quart.), 141.8 (quart.), 133.3 (CH), 123.8 (CH), 123.0 (CH),
121.0 (CH), 112.3 (quart.), 58.8 (CH2(3″)), 58.3 (CH2(5″)),
58.2 (CH2(4″)), 52.0 (CH2(1″)), 51.9 (CH2(2″)), 48.5
(CH2(1′)), 38.9 (CH2(8′)), 29.1 (CH2), 28.2 (CH2), 28.0
(CH2), 27.9 (CH2), 25.8 (CH2), 25.6 (CH2); MS (ESI, 4.1 kV,
8 µl/min, N2, methanol): m/z = 594 (100%) [M – H]–; HRMS
(C31H41N5O7): 595.30060; found: 595.30061.
Following GP4 from 37 (620 mg, 0.95 mmol) 47 (448 mg,
83%) was obtained as an orange-coloured, amorphous solid.
UV-vis (methanol): kmax (log e) = 283 nm (4.53); IR (KBr):
m = 3423s, 2934m, 1590s, 1408w, 1336m, 1260m, 1171w,
1122w cm–1; 1H NMR (500 MHz, D2O): d = 7.51 (d,
3JH,H = 8.71 Hz, 2 H, H-C(4,5)), 7.42 (dd, 3JH,H = 8.71 Hz,
3
3JH,H = 7.33 Hz, 2 H, H-C(3,6)), 7.11 (d, JH,H = 8.71 Hz,
2 H, H-C(1,8)), 7.01 (dd, 3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz,
2 H, CH(2,7)), 3.29–3.24 (m, 2 H, CH2(1′)), 3.09 (s, 4 H, 2 ×
CH2(5″)), 3.07 (s, 2 H, CH2(4″)), 3.06–3.02 (m, 2 H,
CH2(6′)), 3.02 (s, 2 H, CH2(3″)), 2.60–2.53 (m, 4 H, CH2(1″,
2″)), 1.45–1.37 (m, 2 H, CH2(2′)), 1.35–1.31 (m, 2 H,
CH2(5′)), 1.15–1.09 (m, 4 H, 2 × CH2(3′,4′)); 13C NMR
(125 MHz, D2O): d = 179.1 (C=O), 173.8 (C=O), 154.2
(quart.), 141.5 (quart.), 133.3 (CH), 123.8 (CH), 123.0 (CH),
120.7 (CH), 112.1 (quart.), 58.8 (CH2(3″)), 58.4 (CH2(5″)),
58.2 (CH2(4″)), 52.0 (CH2(1″)), 51.9 (CH2(2″)), 48.4
(CH2(1′)), 38.8 (CH2(6′)), 29.1 (CH2), 28.1 (CH2), 25.5
(CH2), 25.4 (CH2); MS (ESI, 4.1 kV, 8 µl/min, N2, metha-
nol): m/z = 566 (100%) [M – H]–; HRMS (C29H37N5O7):
567.26930; found: 567.26931.
5.2.37. 2-[{2-[(2-{[9-(9-Acridinylamino)nonyl]amino}-2-
oxoethyl)(carboxymethyl)- amino]ethyl}(carboxy-
methyl)amino] acetic acid (50)
Following GP4 from 40 (730 mg, 1.05 mmol) 50 (564 mg,
88%) was obtained as a yellow, amorphous solid. UV-vis
(methanol): kmax(log e) = 284 nm (4.55); IR (KBr):
1
m = 3442s, 1591s, 1408m, 1337w, 1124w cm–1; H NMR
3
(500 MHz, D2O): d = 7.42 (d, JH,H = 8.68 Hz, 2 H,
5.2.35. 2-[{2-[(2-{[7-(9-Acridinylamino)heptyl]amino}-2-
oxoethyl)(carboxymethyl)- amino]ethyl}(carboxy-
methyl)amino] acetic acid (48)
H-C(4,5)), 7.36 (dd, 3JH,H = 8.68 Hz, 3JH,H = 7.63 Hz, 2 H,
H-C(3,6)), 7.04 (d, 3JH,H = 8.68 Hz, 2 H, H-C(1,8)), 6.92 (dd,
3JH,H = 8.68 Hz, 3JH,H = 7.63 Hz, 2 H, H-C(2,7)), 3.38 (s, 4 H,
2 × CH2(5″)), 3.20–3.15 (m, 2 H, CH2(1′)), 3.17 (s, 2 H,
CH2(4″)), 3.07 (s, 2 H, CH2(3″)), 2.99–2.96 (m, 2 H,
CH2(9′)), 2.91–2.85 (m, 2 H, CH2(1″)), 2.75–2.70 (m, 2 H,
CH2(2″)), 1.31–1.20 (m, 4 H, 2 × CH2(2′,8′)), 0.98–0.85 (m,
10 H, 5 × CH2(3′,4′,5′,6′,7′)); 13C NMR (125 MHz, D2O):
d = 178.6 (C=O), 173.2 (C=O), 154.9 (quart.), 139.5 (quart.),
134.2 (CH), 123.8 (CH), 123.3 (CH), 119.1 (CH), 111.3
(quart.), 58.5 (CH2(3″)), 58.3 (CH2(5″)), 57.6 (CH2(4″)),
52.3 (CH2(1″)), 51.0 (CH2(2″)), 48.2 (CH2(1′)), 39.1
(CH2(9′)), 28.9 (CH2), 28.4 (CH2), 28.3 (CH2), 28.2 (CH2),
28.0 (CH2), 26.1 (CH2), 25.7 (CH2); MS (ESI, 4.1 kV,
8 µl/min, N2, methanol): m/z = 608 (100%) [M –H]–; HRMS
(C32H43N5O7): 609.31625; found: 609.31625.
Following GP4 from 38 (250 mg, 0375 mmol) 48
(172 mg, 79%) was obtained as an orange-coloured, amor-
phous solid. UV-vis (methanol): kmax (log e) = 281 nm
(4.68); IR (KBr): m = 3425s, 2935w, 2856w, 1635s, 1592s,
1
1473w, 1407m, 1338w, 1272w, 1171w, 1123w cm–1; H
3
NMR (500 MHz, D2O): d = 7.28 (dd, JH,H = 8.71 Hz,
3
3JH,H = 7.33 Hz, 2 H, H-C(3,6)), 7.25 (d, JH,H = 8.71 Hz,
2 H, H-C(4,5)), 6.85 (dd, 3JH,H = 8.71 Hz, 3JH,H = 7.33 Hz,
2 H, H-C(2,7)), 6.83 (d, 3JH,H = 8.71 Hz, 2 H, H-C(1,8)), 3,34
(s, 4 H, 2 × CH2(5″)), 3.17 (s, 2 H, CH2(4″)), 3.06 (s, 2 H,
CH2(3″)), 3.07–3.01 (m, 4 H, 2 × CH2(1′,7′)), 2.88–2.80 (s,
2 H, CH2(1″)), 2.74–2.68 (s, 2 H, CH2(2″)), 1.35–1.22 (m,
4 H, 2 × CH2(2′,6′)), 1.09–1.00 (m, 6H, 3xCH2(3′,4′,5′)); 13
C
NMR (125 MHz, D2O): d = 178.7 (C=O), 173.5 (C=O),
154.3 (quart.), 139.1 (quart.), 134.1 (CH), 123.6 (CH), 123.2
(CH), 118.8 (CH), 110.9 (quart.), 58.6 (CH2(3″)), 58.3
(CH2(5″)), 57.7 (CH2(4″)), 52.2 (CH2(1″)), 51.4 (CH2(2″)),
48.1 (CH2(1′)), 38.9 (CH2(7′)), 28.8 (CH2), 28.2 (CH2), 27.7
(CH2), 25.8 (CH2), 25.6 (CH2); MS (ESI, 4.1 kV, 8 µl/min,
N2, methanol): m/z = 580 (100%) [M – H]–; HRMS
(C30H39N5O7): 581.28495; found: 581.28497.
5.2.38. 2-[{2-[(2-{[10-(9-Acridinylamino)decyl]amino}-2-
oxoethyl)(carboxymethyl)- amino]ethyl}(carboxy-
methyl)amino] acetic acid (51)
Following GP4 from 41 (260 mg, 0.367 mmol) 51
(174 mg, 76%) was obtained as an amorphous, yellow solid.
UV-vis (methanol): kmax (log e) = 284 nm (4.66); IR (KBr):

988
R. Csuk et al. / European Journal of Medicinal Chemistry 39 (2004) 975–988
m = 3418s, 2924m, 2851s, 1644s, 1590s, 1473m, 1408m,
References
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3
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52.2 (CH2(1″)), 51.3 (CH2(2″)), 48.4 (CH2(1′)), 39.1
(CH2(10′)), 29.1 (CH2), 28.60 (CH2), 28.56 (CH2), 28.40
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3
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Acknowledgements
Thanks for helpful discussion and parts of the biological
screening are due to Dr. H. Knoller and Dr. H.-J. Neumann
(Fresenius Hemocare); additional screening as been perfor-
med by Bioscreen Ltd. Financial support by the European
Communities (SC1*-CT92-0780) and the Fonds der Che-
mischen Industrie is gratefully acknowledged. We like to
thank Dr. R. Kluge for the ESI-MS spectra and Dr. D. Ströhl
for taking numerous NMR spectra.