Synthesis of dimeric trifluoromethoxyacridine-derived pathogen-inactivating nucleic acid intercalators

Article abstract of DOI:10.1002/ardp.200400907

A series of antiviral active compounds consisting of an intercalating acridine-derived part, a spacer region and a reactive EDTA-derived conjugate was synthesized in an easy sequence. Thus, suitably mono-protected 1,ω-alkyldiamines gave, upon reaction with 9-chloro-2- trifluoromethoxyacridine, followed by deprotection and reaction with EDTA dianhydride, the target molecules. Incorporation of their Fe(II) complexes in the presence of ascorbate gave a reduction of the phage titer of MS2 phages by several logarithmic decades.

Full text of DOI:10.1002/ardp.200400907

Arch. Pharm. Pharm. Med. Chem. 2004, 337, 571−578
Pathogen-inactivating Intercalators 571
´
Rene Csuk,
Synthesis of Dimeric Trifluoromethoxyacridine-
derived Pathogen-inactivating Nucleic Acid
Intercalators
Christian Raschke,
Gunnar Göthe,
Stefan Reißmann
Institut für Organische Chemie,
Martin-Luther-Universität
Halle-Wittenberg,
D-06120 Halle (Saale),
Germany
A series of antiviral active compounds consisting of an intercalating acridine-
derived part, a spacer region and a reactive EDTA-derived conjugate was syn-
thesized in an easy sequence. Thus, suitably mono-protected 1,ω-alkyldiamines
gave, upon reaction with 9-chloro-2-trifluoromethoxyacridine, followed by depro-
tection and reaction with EDTA dianhydride, the target molecules. Incorporation
of their Fe(II) complexes in the presence of ascorbate gave a reduction of the
phage titer of MS2 phages by several logarithmic decades.
Keywords: Pathogen Inactivation; Nucleic Acid Intercalators; Acridine
Received: June 11, 2004; accepted: September 2, 2004 [FP907]
DOI 10.1002/ardp.200400907
Introduction
The safety of blood and plasma is the subject of an
ongoing concern among the public, underlining the
need for the development of efficient analytical sys-
tems as well as of anti-pathogenic drugs. Life-threat-
ening diseases can be the result of accepting blood or
plasma donations if they carry viral infectious agents.
The potential bioburden of the donated blood has to
be minimized, but this goal cannot completely be
achieved by simple testing of blood and plasma do-
nations for the presence of infectious agents, since it
is not possible to perform assays against all known
and unknown biological threats. Thus, the develop-
ment of anti-pathogenic measures seems to be
called for.
Figure 1. Types of anti-pathogenic compounds.
the damage to the biological molecules. Thus, the re-
activity of the molecules can be triggered by the pres-
ence and the concentration of a more or less harm-
less agent.
Results and discussion
Quite recently [1], a new type of pathogen-inactivating
agents (type A; Figure 1) was introduced, consisting
of an intercalator that binds to the nucleic acid of
pathogens combined with a conjugate that sub-
sequently destroys the nucleic acid via a Fe(II)-me-
diated Fenton mechanism. The latter process ac-
complishes damage to the nucleic acids by producing
OH radicals. Since the metal attached to the conjugate
can change between two levels of oxidation, a cycle
can finally be constructed in the presence of suitable
reduction agents (e.g. ascorbic acid) that increases
Of paramount importance is the interaction of the inter-
calator with the nucleic acids. Thus, it can safely be
assumed that dimeric structures (type B; Figure 1)
should provide a better intercalation and thus a higher
anti-pathogenic activity. From preliminary modelling
studies we deduced that the presence of a 2-trifluor-
methoxy moiety in the acridine ring should result in an
improved interaction between the intercalator and the
nucleic acids.
From a palladium-assisted Buchwald-Hartwig ami-
nation reaction between methyl 2-iodobenzoate and
4-trifluoromethoxyaniline [2], methyl 2-[4-(trifluoro-
methoxy)amino]benzoate [3] was obtained. Saponi-
fication followed by a POCl3-mediated cyclization
furnished the starting material for our sequence,
´
Correspondence: Rene Csuk, Institut für Organische
Chemie, Martin-Luther-Universität Halle-Wittenberg, Kurt-
Mothes-Straße 2, D-06120 Halle (Saale), Germany;
Phone: +49 345 5525660, Fax: +49 345 5527030, e-mail:
csuk@chemie.uni-halle.de
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

572 Csuk et al.
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 571−578
Scheme 1. Syntheses of the target molecules. Reagents: a) phenol; b) aq. HCl; c) EDTA dianhydride.
9-chloro-2-trifluoromethoxyacridine (1) (Scheme 1).
Reaction of 1 with monoprotected diamines, 1-amino-
x-tert-butyloxycarbonylaminoalkanes 2 [4], 3 [5], 4 [6],
5 [7], 6 [8], 7 [9] or 8 [10] in phenol [11, 12] gave the
acridinyl carbamates 9Ϫ15, respectively. These acrid-
inyl carbamates were deprotected using aqueous hy-
drochloric acid to afford the corresponding bishydro-
chlorides 16Ϫ22. These compounds were allowed to
react with EDTA dianhydride to yield the target
compounds 23؊29 (Scheme 1).
Acknowledgments
Thanks for helpful discussion and parts of the biologi-
cal screening are due to Dr. H. Knoller and Dr. H.-J.
For biological screening, the compounds were loaded
with three equivalents of Fe(III) and then incubated in
Tris buffer with MS2 phages [13], in the presence of
sodium ascorbate. Whereas almost no anti-patho-
genic acivity was observed in the absence of ascor-
bate, in the presence of ascorbic acid the phage titer
of the MS2 phages was reduced by several logarith-
mic decades (Figure 2). As compared to monomeric,
unsubstituted acridine analogues [1] or to the photo-
inactivation by porphyrins [14], an approximately 100-
fold higher anti-pathogenic activity could be achieved.
Modifications in the structure of these compounds, as
well as an extended biological screening, are pre-
sently performed in our laboratories.
Figure 2. Inactivation of phage MS2 as a function of
spacer length [20°C, 5 mmol Na-ascorbate, 100 µM of
compounds 23؊29 (Fe3+ loaded, 3 equiv.)].
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Arch. Pharm. Pharm. Med. Chem. 2004, 337, 571−578
Pathogen-inactivating Intercalators 573
for 30 min; 2 (1.18 g, 7.4 mmol) was added, and stirring at
this temperature continued for another 15 min. The product
was purified by chromatography (silica gel, methanol/ethyl
acetate 1 : 4) to afford 9 (1.47 g, 52%) as a red, highly vis-
cous oil. UV-vis (methanol): λmax (log ε) = 284 nm (4.68).
IR (film): ν = 3273w, 2981m, 1703m, 1652m, 1634m, 1592s,
1533s, 1407m, 1368m, 1334w, 1261s, 1215s, 1166s, 1044w,
Neumann (Fresenius Hemocare); additional screening
has been performed by Bioscreen Ltd. Financial sup-
port, by the European Communities (SC1-CT92-0780)
and the Fonds der Chemischen Industrie, is gratefully
acknowledged. We like to thank Dr. D. Ströhl for nu-
merous NMR spectra, Dr. R. Kluge for the MS spectra,
Mr. T. Brezesinski and Ms. B. Janeska for her help in
the preparation of starting materials.
1012w cmϪ1. 1H NMR (400 MHz, CDCl3): δ = 8.14 (d, 3JH,H
=
=
3
9.55 Hz, 1H, H-C(4)), 8.12 (s, 1H, H-C(1)), 7.91 (d, JH,H
3
8.72 Hz, 1H, H-C(8)), 7.75 (d, JH,H = 8.72 Hz, 1H, H-C(5)),
3
7.41Ϫ7.36 (m, 2H, H-C(3,6)), 7.11 (dd, JH,H = 8.72 Hz,
3JH,H = 6.74 Hz, 1H, H-C(7)), 6.33 (br s, 1H, NH), 4.02Ϫ3.98
(m, 2H, CH2(1Ј)), 3.68Ϫ3.62 (m, 2H, CH2(2Ј)), 1.45 (s, 9H,
tBu). 13C NMR (100 MHz, CDCl3): δ = 157.8 (quart.), 155.1
(C=O), 143.8 (quart.), 142.0 (quart.), 140.1 (quart.), 132.9
(CH), 127.0 (CH), 124.6 (CH), 123.8 (CH), 123.2 (CH), 121.6
Experimental
General
1
(CH), 120.5 (q, JC,F = 257.4 Hz, OCF3), 116.2 (CH), 113.0
Melting points are uncorrected (Leica hot-stage microscope).
NMR spectra were recorded using the Varian spectrometers
Gemini 200, Gemini 2000 or Unity 500 (δ given in ppm, J
inHz, internal Me4Si); IR spectra (film or KBr pellet) were re-
corded 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
instrument (electrospray, voltage 4.5 kV, sheath gas nitro-
gen). For elemental analysis, a Foss-Heraeus Vario EL instru-
ment was used. TLC was performed on silica gel (Merck
5554); detection by UV absorption or by treatment with a
solution of 10% sulfuric acid, ammonium molybdate and ce-
rium(IV) sulfate, followed by gentle heating. The solvents were
dried according to usual procedures.
(quart.), 112.8 (quart.), 80.4 (quart., tBu), 51.2 (CH2(1Ј)), 40.4
(CH2(2Ј)), 28.4 (CH3, Bu). 19F NMR (188 MHz, CDCl3): δ =
t
Ϫ58.7 (OCF3). MS (ESI, 4.1 kV, 8 µL/min, N2, methanol): 366
(10%) [(M-ibutene)H]+, 422 (100%) [MH]+. Analysis for
C21H22F3N3O3 (421.42): C 59.85; H 5.26; N 9.97; found: C
59.64; H 5.37; N 9.81.
tert-Butyl N-{3-[9-(2-trifluoromethoxyacridinyl)amino]propyl}
carbamate (10)
As described for 9, from 1 (2.0 g, 6.7 mmol), phenol (10.0 g,
0.1 mol) and 3 (1.42 g, 8.2 mmol), 10 (1.23 g, 42%) was
obtained as a red, highly viscous oil. UV-vis (methanol): λmax
(log ε) = 284 nm (4.71). IR (film): ν = 3441s, 2980m, 1690m,
1630m, 1568s, 1524m, 1444w, 1392w, 1368m, 1336w,
1286s, 1262s, 1216m, 1169m, 1084w, 1020w cmϪ1. 1H NMR
Bacteriophages and screening
3
(400 MHz, CDCl3): δ = 8.21 (d, JH,H = 9.55 Hz, 1H, H-C(4)),
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-contain-
ing 0.0025% CaCl2. The cultures were incubated in a 37°C
shaking water bath for 4 h, until the log phase was reached.
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 it was incubated overnight at
37°C. The resulting 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 ϫ 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 ϫ 109 pfu/mL.
Dilutions were prepared in phosphate-buffered saline solu-
tion. Initial as well as 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.
3
8.10 (s, 1H, H-C(1)), 8.08 (d, JH,H = 8.72 Hz, 1H, H-C(8)),
3
3
8.07 (d, JH,H = 8.72 Hz, 1H, H-C(5)), 7.66 (dd, JH,H = 8.72
3
3
Hz, JH,H = 6.64 Hz, 1H, H-C(6)), 7.52 (dd, JH,H = 9.55 Hz,
4JH,H = 2.08 Hz, 1H, H-C(3)), 7.39 (dd, JH,H = 8.72 Hz,
3
3JH,H = 6.64 Hz, 1H, H-C(7)), 6.55 (br s, 1H, NH), 4.78 (br
s, 1H, NH), 3.78Ϫ3.75 (m, 2H, CH2(1Ј)), 3.44Ϫ3.41 (m, 2H,
CH2(3Ј)), 1.92Ϫ1.86 (m, 2H, CH2(2Ј)), 1.50 (s, 9H, Bu). 13C
t
NMR (100 MHz, CDCl3): δ = 157.2 (quart.), 152.1 (C=O),
148.9 (quart.), 147.5 (quart.), 144.1 (quart.), 130.1 (2 ϫ CH),
1
124.5 (2 ϫ CH), 123.5 (CH), 122.8 (CH), 120.5 (q, JC,F
=
256.7 Hz, OCF3), 117.1 (quart.), 116.5 (quart.), 114.2 (CH),
80.1 (quart., tBu), 46.5 (CH2(1Ј)), 37.4 (CH2(3Ј)), 32.1
t
(CH2(2Ј)), 28.4 (CH3, Bu). 19F NMR (188 MHz, CDCl3): δ =
Ϫ58.5 (OCF3). MS (ESI, 4.1 kV, 8 µL/min, N2, methanol): 380
(20%) [(M-ibutene)H]+, 436 (100%) [MH]+. Analysis for
C22H22F3N3O3 (435.45): C 60.68; H 5.56; N 9.65; found: C
60.41; H 5.74; N 9.51.
tert-Butyl
N-{4-[9-(2-trifluoromethoxyacridinyl)amino]butyl}
carbamate (11)
As described for 9, from 1 (2.0 g, 6.7 mmol), phenol (10.0 g,
0.1 mol) and 4 (1.26 g, 6.7 mmol), 11 (2.09 g, 69%) was
obtained as a red, highly viscous oil. UV-vis (methanol): λmax
(log ε) = 286 nm (4.61). IR (film): ν = 3380m, 2979m, 1699s,
1649m, 1596s, 1535s, 1407m, 1366m, 1260s, 1212s, 1162s,
1005m cmϪ1. 1H NMR (400 MHz, CDCl3): δ = 8.04 (d, 3JH,H
=
=
3
8.72 Hz, 1H, H-C(8)), 7.96 (s, 1H, H-C(1)), 7.83 (d, JH,H
3
9.55 Hz, 1H, H-C(4)), 7.74 (d, JH,H = 8.72 Hz, 1H, H-C(5)),
tert-Butyl
N-{2-[9-(2-trifluoromethoxyacridinyl)amino]ethyl}
3
3
7.45 (dd, JH,H = 8.72 Hz, JH,H = 6.64 Hz, 1H, H-C(6)), 7.36
carbamate (9)
3
4
(dd, JH,H = 9.55 Hz, JH,H = 2.08 Hz, 1H, H-C(3)), 7.15 (dd,
3
A mixture of 9-chloro-2-trifluoromethoxyacridine (1) (2.00 g,
6.7 mmol) and phenol (10.0 g, 0.1 mol) was stirred at 90°C
3JH,H = 8.72 Hz, JH,H = 6.64 Hz, 1H, H-C(7)), 4.93 (br s, 1H,
NH), 3.84Ϫ3.79 (m, 2H, CH2(1Ј)), 3.23Ϫ3.16 (m, 2H,
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

574 Csuk et al.
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 571−578
CH2(4Ј)), 1.97Ϫ1.88 (m, 2H, CH2(2Ј)), 1.71Ϫ1.63 (m, 2H,
(OCF3). MS (ESI, 4.1 kV, 8 µL/min, N2, methanol): 378 (4%)
[(M-Boc)H]+, 422 (12%) [(M-ibutene)H]+, 478 (100%) [MH]+.
Analysis for C25H30F3N3O3 (477.53): C 62.88; H 6.33; N 8.80;
found: C 62.71; H 6.43; N 8.75.
t
CH2(3Ј)), 1.43 (s, 9H, Bu). 13C NMR (100 MHz, CDCl3): δ =
156.2 (quart.), 155.3 (C=O), 143.5 (quart.), 140.7 (quart.),
139.3 (quart.), 133.2 (CH), 127.1 (CH), 124.6 (CH), 123.1
1
(CH), 122.6 (CH), 120.5 (q, JC,F = 258.6 Hz, OCF3), 120.4
(CH), 116.6 (CH), 112.6 (quart.), 112.3 (quart.), 79.2 (quart.,
tBu), 48.3 (CH2(1Ј)), 39.9 (CH2(4Ј)), 28.4 (CH3, Bu), 27.4
t
tert-Butyl N-{7-[9-(2-trifluoromethoxyacridinyl)amino]heptyl}
carbamate (14)
(CH2), 27.2 (CH2). 19F NMR (188 MHz, CDCl3): δ = Ϫ58.7
(OCF3). MS (ESI, 4.1 kV, 8 µL/min, N2, methanol): 394 (14%)
[(M-ibutene)H]+, 450 (100%) [MH]+. Analysis for
C23H26F3N3O3 (449.48): C 61.46; H 5.83; N 9.35; found: C
61.32; H 5.99; N 9.21.
As described for 9, from 1 (2.0 g, 6.7 mmol), phenol (10.0 g,
0.1 mol) and 7 (1.70 g, 7.4 mmol), 14 (1.59 g, 48%) was
obtained as a red, highly viscous oil. UV-vis (methanol): λmax
(log ε) = 286 nm (4.71). IR (film): ν = 3425s, 2930s, 2856s,
1690s, 1632s, 1591s, 1566s, 1528s, 1502s, 1391m, 1365m,
tert-Butyl N-{5-[9-(2-trifluoromethoxyacridinyl)amino]pentyl}
carbamate (12)
1276m, 1247s, 1170s, 1030m cmϪ1
.
1H NMR (400 MHz,
3
CDCl3): δ = 8.09 (d, JH,H = 9.55 Hz, 1H, H-C(4)), 8.06 (d,
3
As described for 9, from 1 (2.0 g, 6.7 mmol), phenol (10.0 g,
0.1 mol) and 5 (1.49 g, 7.4 mmol), 12 (2.12 g, 68%) was
obtained as a red, highly viscous oil. UV-vis (methanol): λmax
(log ε) = 282 nm (4.69). IR (film): ν = 3439s, 2934w, 1630m,
3JH,H = 8.72 Hz, 1H, H-C(8)), 8.03 (d, JH,H = 8.72 Hz, 1H,
4
H-C(5)), 7.92 (d, JH,H = 2.08 Hz, 1H, H-C(1)), 7.68 (dd,
3
3JH,H = 8.72 Hz, JH,H = 6.64 Hz, 1H, H-C(6)), 7.52 (dd,
4
3JH,H = 9.55 Hz, JH,H = 2.08 Hz, 1H, H-C(3)), 7.41 (dd,
1508w, 1367w, 1261s, 1218m, 1165m cmϪ1
.
1H NMR (400
3JH,H = 8.72 Hz, JH,H = 6.64 Hz, 1H, H-C(7)), 5.00 (br s,
3
3
MHz, CDCl3): δ = 8.07 (d, JH,H = 9.55 Hz, 1H, H-C(4)), 8.05
1H, NH), 4.47 (br s, 1H, NH), 3.80Ϫ3.75 (m, 2H, CH2(1Ј)),
3.11Ϫ3.03 (m, 2H, CH2(7Ј)), 1.81Ϫ1.73 (m, 2H, CH2(2Ј)),
(d, 3JH,H = 8.72 Hz, 1H, H-C(8)), 8.03 (d, 3JH,H = 8.72 Hz, 1H,
4
t
H-C(5)), 7.92 (d, JH,H = 2.08 Hz, 1H, H-C(1)), 7.67 (dd,
1.47Ϫ1.25 (m, 17H, Bu, 4 ϫ CH2(3ЈϪ6Ј)). 13C NMR (100
3
3JH,H = 8.72 Hz, JH,H = 6.64 Hz, 1H, H-C(6)), 7.51 (dd,
MHz, CDCl3): δ = 155.9 (C=O), 151.2 (quart.), 149.3 (quart.),
4
3JH,H = 9.55 Hz, JH,H = 2.08 Hz, 1H, H-C(3)), 7.40 (dd,
147.7 (quart.), 144.0 (quart.), 131.9 (CH), 130.1 (CH), 129.8
3
1
3JH,H = 8.72 Hz, JH,H = 6.64 Hz, 1H, H-C(7)), 5.05 (br s,
(CH), 124.4 (CH), 123.9 (CH), 121.8 (CH), 120.5 (q, JC,F
=
1H, NH), 4.53 (br s, 1H, NH), 3.79Ϫ3.74 (m, 2H, CH2(1Ј)),
3.15Ϫ3.08 (m, 2H, CH2(5Ј)), 1.84Ϫ1.76 (m, 2H, CH2(2Ј)),
256.4 Hz, OCF3), 116.7 (quart.), 115.9 (quart.), 113.8 (CH),
t
79.0 (quart., Bu), 50.8 (CH2(1Ј)), 40.5 (CH2(7Ј)), 31.7 (CH2),
t
t
1.54Ϫ1.42 (m, 4H, 2 ϫ CH2(3Ј,4Ј)), 1.41 (s, 9H, Bu). 13C
30.0 (CH2), 29.0 (CH2), 28.5 (CH3, Bu), 26.8 (CH2), 26.6
NMR (100 MHz, CDCl3): δ = 155.9 (C=O), 151.2 (quart.),
149.1 (quart.), 147.5 (quart.), 144.0 (quart.), 131.6 (CH),
130.1 (CH), 129.4 (CH), 124.4 (CH), 123.9 (CH), 121.9 (CH),
(CH2). 19F NMR (188 MHz, CDCl3): δ = Ϫ58.5 (OCF3). MS
(ESI, 4.1 kV, 8 µL/min, N2, methanol): 392 (8%) [(M-Boc)H]+,
436 (15%) [(M-ibutene)H]+, 492 (100%) [MH]+. Analysis for
C26H32F3N3O3 (491.56): C 63.53; H 6.56; N 8.55; found: C
63.41; H 6.71; N 8.39.
1
120.5 (q, JC,F = 256.6 Hz, OCF3), 116.7 (quart.), 115.9
t
(quart.), 113.9 (CH), 79.2 (quart., Bu), 50.7 (CH2(1Ј)), 40.2
(CH2(5Ј)), 31.3 (CH2), 29.9 (CH2), 28.4 (CH3, tBu), 24.1
(CH2). 19F NMR (188 MHz, CDCl3): δ = Ϫ58.5 (OCF3). MS
(ESI, 4.1 kV, 8 µL/min, N2, methanol): 408 (12%) [(M-
ibutene)H]+, 464 (100%) [MH]+. Analysis for C24H28F3N3O3
(463.50): C 62.19; H 6.09; N 9.07; found: C 62.00; H 6.23;
N 8.95.
tert-Butyl
N-{8-[9-(2-trifluoromethoxyacridinyl)amino]octyl}
carbamate (15)
As described for 9, from 1 (2.0 g, 6.7 mmol), phenol (10.0 g,
0.1 mol) and 8 (2.71 g, 11.1 mmol), 15 (3.09 g, 61%) was
obtained as a red, highly viscous oil. UV-vis (methanol): λmax
(log ε) = 286 nm (4.76). IR (film): ν = 3346m, 2930s, 2857m,
1694s, 1633m, 1564s, 1520s, 1392m, 1366s, 1255s, 1165s,
tert-Butyl
N-{6-[9-(2-trifluoromethoxyacridinyl)amino]hexyl}
carbamate (13)
As described for 9, from 1 (2.0 g, 6.7 mmol), phenol (10.0 g,
0.1 mol) and 6 (1.60 g, 7.4 mmol), 13 (1.24 g, 39%) was
obtained as a red, highly viscous oil. UV-vis (methanol): λmax
(log ε) = 287 nm (4.70). IR (film): ν = 3358m, 3009m, 2934s,
2860m, 1692s, 1619m, 1561s, 1509s, 1476s, 1392s, 1366s,
1041m cmϪ1. 1H NMR (400 MHz, CDCl3): δ = 8.05 (d, 3JH,H
9.55 Hz, 1H, H-C(4)), 8.03 (d, JH,H = 8.72 Hz, 1H, H-C(8)),
=
3
3
4
8.02 (d, JH,H = 8.72 Hz, 1H, H-C(5)), 7.92 (d, JH,H = 2.08
3
3
Hz, 1H, H-C(1)), 7.65 (dd, JH,H = 8.72 Hz, JH,H = 6.64 Hz,
1H, H-C(6)), 7.50 (dd, JH,H = 9.55 Hz, JH,H = 2.08 Hz, 1H,
H-C(3)), 7.38 (dd, JH,H = 8.72 Hz, JH,H = 6.64 Hz, 1H, H-
C(7)), 4.49 (br s, 1H, NH), 3.80Ϫ3.75 (m, 2H, CH2(1Ј)),
3.10Ϫ3.03 (m, 2H, CH2(8Ј)), 1.82Ϫ1.74 (m, 2H, CH2(2Ј)),
3
4
1
3
3
1258s, 1167s, 1022m cmϪ1. H NMR (400 MHz, CDCl3): δ =
3
3
8.08 (d, JH,H = 9.55 Hz, 1H, H-C(4)), 8.06 (d, JH,H = 8.72
3
Hz, 1H, H-C(8)), 8.05 (d, JH,H = 8.72 Hz, 1H, H-C(5)), 7.94
(s, 1H, H-C(1)), 7.67 (dd, JH,H = 8.72 Hz, JH,H = 6.64 Hz,
1H, H-C(6)), 7.51 (dd, JH,H = 9.55 Hz, JH,H = 2.08 Hz, 1H,
H-C(3)), 7.39 (dd, JH,H = 8.72 Hz, JH,H = 6.64 Hz, 1H, H-
C(7)), 5.15 (br s, 1H, NH), 4.51 (br s, 1H, NH), 3.79Ϫ3.74
(m, 2H, CH2(1Ј)), 3.13Ϫ3.05 (m, 2H, CH2(6Ј)), 1.81Ϫ1.73 (m,
3
3
t
1.46Ϫ1.38 (m, 13H, Bu, 2 ϫ CH2(3Ј,7Ј)), 1.33Ϫ1.23 (m, 6H,
3
4
3
CH2(4Ј,5Ј,6Ј)). 13C NMR (100 MHz, CDCl3): δ = 155.9
3
3
(C=O), 151.2 (quart.), 149.3 (quart.), 147.6 (quart.), 144.0
(quart.), 131.8 (CH), 130.1 (CH), 129.7 (CH), 124.5 (CH),
1
123.9 (CH), 121.8 (CH), 120.7 (q, JC,F = 256.7 Hz, OCF3),
t
t
2H, CH2(2Ј)), 1.50Ϫ1.32 (m, 15H, Bu, 3 ϫ CH2(3Ј,4Ј,5Ј)).
116.7 (quart.), 115.9 (quart.), 113.8 (CH), 79.0 (quart., Bu),
13C NMR (100 MHz, CDCl3): δ = 155.9 (C=O), 151.2 (quart.),
149.2 (quart.), 147.5 (quart.), 143.9 (quart.), 131.6 (CH),
130.2 (CH), 129.5 (CH), 124.4 (CH), 123.8 (CH), 121.9 (CH),
50.8 (CH2(1Ј)), 40.6 (CH2(8Ј)), 31.7 (CH2), 30.1 (CH2), 29.2
t
(CH2), 29.1 (CH2), 28.5 (CH3, Bu), 26.8 (CH2), 26.7 (CH2).
19F NMR (188 MHz, CDCl3): δ = Ϫ58.5 (OCF3). MS (ESI, 4.1
kV, 8 µL/min, N2, methanol): 406 (4%) [(M-Boc)H]+, 450
(10%) [(M-ibutene)H]+, 506 (100%) [MH]+. Analysis for
C28H34F3N3O3 (517.60): C 64.98; H 6.62; N 8.12; found: C
64.81; H 6.85; N 7.99.
1
120.6 (q, JC,F = 254.6 Hz, OCF3), 116.8 (quart.), 115.8
t
(quart.), 113.9 (CH), 79.1 (quart., Bu), 50.5 (CH2(1Ј)), 40.2
(CH2(6Ј)), 31.6 (CH2), 30.1 (CH2), 28.5 (CH3, tBu), 26.4
(CH2), 26.3 (CH2). 19F NMR (188 MHz, CDCl3): δ = Ϫ58.5
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Arch. Pharm. Pharm. Med. Chem. 2004, 337, 571−578
Pathogen-inactivating Intercalators 575
N1-[9-(2-Trifluoromethoxyacridinyl)-ethane-1,2-diamine bis-
hydrochloride (16)
(188 MHz, D2O): δ = Ϫ58.8 (OCF3). MS (ESI, 4.1 kV, 8 µL/
min, N2, methanol): 350 (100%) [MH]+.
A solution of 9 (1.41 g, 3.3 mmol) in methanol (50 mL) con-
taining aq. hydrochloric acid (10%, 25 mL) was stirred at
room temperature for 16 h. The solvents were removed under
reduced pressure, and 16 (1.29 g, 98%) was obtained as a
yellow, amorphous solid. UV-vis (methanol): λmax (log ε) =
284 nm (4.59). IR (KBr): ν = 3423s, 1592m, 1534m, 1474m,
N1-[9-(2-Trifluoromethoxyacridinyl)]-pentane-1,5-diamine bis-
hydrochloride (19)
As described for 9, from 12 (2.02 g, 4.4 mmol), 19 (1.86 g,
98%) was obtained as yellow, amorphous solid. UV-vis
(methanol): λmax (log ε) = 284 nm (4.49). IR (KBr): ν = 3418s,
2966s, 1629m, 1594s, 1538m, 1485s, 1363w, 1258s, 1207s,
1
1259m, 1165m cmϪ1. H NMR (400 MHz, D2O): δ = 8.12 (d,
4
3JH,H = 8.72 Hz, 1H, H-C(8)), 8.07 (d, JH,H = 1.97 Hz, 1H,
1165s cmϪ1. 1H NMR (400 MHz, CD3OD): δ = 8.55 (d, 3JH,H
=
=
3
3
H-C(1)), 7.82 (dd, JH,H = 8.72 Hz, JH,H = 6.95 Hz, 1H, H-
3
C(7)), 7.75 (dd, 3JH,H = 9.34 Hz, 4JH,H = 1.97 Hz, 1H, H-C(3)),
7.61 (d, JH,H = 9.34 Hz, 1H, H-C(4)), 7.53 (d, JH,H = 8.72
8.72 Hz, 1H, H-C(8)), 8.51 (s, 1H, H-C(1)), 7.99 (dd, JH,H
3
3
3
8.72 Hz, JH,H = 6.95 Hz, 1H, H-C(7)), 7.95Ϫ7.92 (m, 2H, 2
3
3
3
H-C(4,5)), 7.85 (d, JH,H = 9.34 Hz, 1H, H-C(3)), 7.60 (dd,
Hz, 1H, H-C(5)), 7.44 (dd, JH,H = 8.72 Hz, JH,H = 6.95 Hz,
1H, H-C(6)), 4.34Ϫ4.29 (m, 2H, CH2(1Ј)), 3.47Ϫ3.43 (m, 2H,
CH2(2Ј)). 13C NMR (100 MHz, D2O): δ = 157.5 (quart.), 144.3
(quart.), 139.3 (quart.), 137.1 (quart.), 136.0 (CH), 129.4
(CH), 124.8 (2 CH), 120.7 (CH), 120.3 (q, 1JC,F = 257 Hz,
OCF3), 118.4 (CH), 116.2 (CH), 112.2 (quart.), 111.5 (quart.),
45.8 (CH2(1Ј)), 38.5 (CH2(2Ј)). 19F NMR (188 MHz, D2O): δ =
Ϫ58.8 (OCF3). MS (ESI, 4.1 kV, 8 µL/min, N2, methanol): 322
(100%) [MH]+.
3JH,H = 8.72 Hz, 3JH,H = 6.95 Hz, 1H, H-C(6)), 4.21Ϫ4.17 (m,
2H, CH2(1Ј)), 2.98Ϫ2.94 (m, 2H, CH2(5Ј)), 2.10Ϫ2.00 (m, 2H,
CH2(2Ј)), 1.81Ϫ1.70 (m, 2H, CH2(4Ј)), 1.61Ϫ1.54 (m, 2H,
CH2(3Ј)). 13C NMR (125 MHz, CD3OD): δ = 159.4 (quart.),
146.1 (quart.), 141.1 (quart.), 138.9 (quart.), 136.9 (CH),
130.3 (CH), 129.9 (CH), 129.2 (CH), 126.3 (CH), 122.0 (q,
1JC,F = 257 Hz, OCF3), 122.1 (CH), 119.7 (CH), 113.2
(quart.), 113.0 (quart.), 50.2 (CH2(1Ј)), 40.5 (CH2(5Ј)), 29.9
(CH2), 28.1 (CH2), 24.7 (CH2). 19F NMR (188 MHz, CD3OD):
δ = Ϫ60.1 (OCF3). MS (ESI, 4.1 kV, 8 µL/min, N2, methanol):
364 (100%) [MH]+.
N1-[9-(2-Trifluoromethoxyacridinyl)]-propane-1,3-diamine bis-
hydrochloride (17)
N1-[9-(2-Trifluoromethoxyacridinyl)]-hexane-1,6-diamine bis-
hydrochloride (20)
As described for 9, from 10 (1.18 g, 2.7 mmol), 17 (1.04 g,
94%) was obtained as a yellow, amorphous solid. UV-vis
(methanol): λmax (log ε) = 280 nm (4.55). IR (KBr): ν = 3418s,
2928s, 1628s, 1594s, 1537m, 1482s, 1400m, 1367m, 1262s,
As described for 9, from 13 (1.19 g, 2.5 mmol), 20 (1.09 g,
98%) was obtained as a yellow, amorphous solid. UV-vis
(methanol): λmax (log ε) = 284 nm (4.60). IR (KBr): ν = 3445s,
1204s, 1165s cmϪ1 1H NMR (400 MHz, D2O): δ = 7.96 (d,
.
3JH,H = 8.72 Hz, 1H, H-C(8)), 7.89 (s, 1H, H-C(1)), 7.70 (dd,
2936s, 1629m, 1595s, 1538m, 1484m, 1363w, 1260s, 1208s,
3JH,H = 8.72 Hz, 3JH,H = 6.95 Hz, 1H, H-C(7)), 7.64 (d, 3JH,H
=
3
1163s cmϪ1
.
1H NMR (400 MHz, D2O): δ = 7.96 (d, JH,H
=
=
3
9.34 Hz, 1H, H-C(3)), 7.40 (d, JH,H = 9.34 Hz, 1H, H-C(4)),
3
8.72 Hz, 1H, H-C(8)), 7.85 (s, 1H, H-C(1)), 7.72 (dd, JH,H
3
3
7.33 (dd, JH,H = 8.72 Hz, JH,H = 6.95 Hz, 1H, H-C(6)), 7.32
3
3
8.72 Hz, JH,H = 6.95 Hz, 1H, H-C(7)), 7.66 (d, JH,H = 9.34
Hz, 1H, H-C(3)), 7.44 (d, JH,H = 9.34 Hz, 1H, H-C(4)), 7.37
(d, 3JH,H = 8.72 Hz, 1H, H-C(5)), 3.95Ϫ3.89 (m, 2H, CH2(1Ј)),
3
3.02Ϫ2.97 (m, 2H, CH2(3Ј)), 2.17Ϫ2.09 (m, 2H, CH2(2Ј)). 13
C
3
3
(d, JH,H = 8.72 Hz, 1H, H-C(5)), 7.33 (dd, JH,H = 8.72 Hz,
3JH,H = 6.95 Hz, 1H, H-C(6)), 3.79Ϫ3.73 (m, 2H, CH2(1Ј)),
2.88Ϫ2.82 (m, 2H, CH2(6Ј)), 1.76Ϫ1.68 (m, 2H, CH2(2Ј)),
1.57Ϫ1.50 (m, 2H, CH2(5Ј)), 1.34Ϫ1.26 (m, 4H, CH2(3Ј,4Ј)).
13C NMR (125 MHz, D2O): δ = 156.5 (quart.), 135.6 (CH),
129.1 (CH), 120.3 (CH), 120.2 (q, 1JC,F = 258 Hz, OCF3),
122.1 (CH), 118.0 (CH), 48.5 (CH2(1Ј)), 39.3 (CH2(6Ј)), 28.7
(CH2), 26.6 (CH2), 25.4 (CH2), 25.2 (CH2). 19F NMR (188
MHz, D2O): δ = Ϫ58.8 (OCF3). MS (ESI, 4.1 kV, 8 µL/min,
N2, methanol): 378 (100%) [MH]+.
NMR (100 MHz, D2O): δ = 157.0 (quart.), 144.0 (quart.),
139.0 (quart.), 137.1 (quart.), 136.8 (CH), 129.3 (CH), 124.5
(CH), 124.1 (CH), 120.5 (CH), 120.3 (q, 1JC,F = 258 Hz,
OCF3), 118.2 (CH), 116.4 (CH), 111.8 (quart.), 111.4 (quart.),
45.9 (CH2(1Ј)), 36.9 (CH2(3Ј)), 27.0 (CH2(2Ј)). 19F NMR (188
MHz, D2O): δ = Ϫ58.8 (OCF3). MS (ESI, 4.1 kV, 8 µL/min,
N2, methanol): 336 (100%) [MH]+.
N1-[9-(2-Trifluoromethoxyacridinyl)]-butane-1,4-diamine bis-
hydrochloride (18)
N1-[9-(2-Trifluoromethoxyacridinyl)]-heptane-1,7-diamine bis-
As described for 9, from 11 (1.50 g, 3.3 mmol), 18 (1.39 g,
99%) was obtained as a yellow, amorphous solid. UV-vis
(methanol): λmax (log ε) = 285 nm (4.52). IR (KBr): ν = 3427s,
hydrochloride (21)
As described for 9, from 14 (1.54 g, 3.1 mmol), 21 (1.41 g,
97%) was obtained as a yellow, amorphous soild. UV-vis
(methanol): λmax (log ε) = 286 nm (4.62). IR (KBr): ν = 3421m,
2929s, 1631s, 1596s, 1535m, 1477m, 1366w, 1261s, 1207s,
3
1162s cmϪ1
.
1H NMR (400 MHz, D2O): δ = 8.08 (d, JH,H
=
=
3
8.72 Hz, 1H, H-C(8)), 7.99 (s, 1H, H-C(1)), 7.77 (dd, JH,H
2930s, 1629m, 1596s, 1540m, 1477m, 1367w, 1257s, 1212s,
8.72 Hz, JH,H = 6.95 Hz, 1H, H-C(7)), 7.70 (d, JH,H = 9.34
Hz, 1H, H-C(3)), 7.55 (d, JH,H = 9.34 Hz, 1H, H-C(4)), 7.48
1156s cmϪ1
.
1H NMR (400 MHz, D2O): δ = 7.95 (d, JH,H
=
=
3
3
3
3
3
8.72 Hz, 1H, H-C(8)), 7.84 (s, 1H, H-C(1)), 7.72 (dd, JH,H
3
3
3
3
(dd, JH,H = 8.72 Hz, JH,H = 6.95 Hz, 1H, H-C(6)), 7.37 (d,
3JH,H = 8.72 Hz, 1H, H-C(5)), 3.95Ϫ3.90 (m, 2H, CH2(1Ј)),
2.92Ϫ2.87 (m, 2H, CH2(4Ј)), 1.86Ϫ1.78 (m, 2H, CH2(2Ј)),
1.68Ϫ1.58 (m, 2H, CH2(3Ј)). 13C NMR (100 MHz, D2O): δ =
155.8 (quart.), 143.8 (quart.), 138.4 (quart.), 136.7 (quart.),
135.7 (CH), 129.1 (CH), 124.6 (CH), 120.3 (CH), 120.3 (q,
1JC,F = 258 Hz, OCF3), 117.8 (CH), 110.8 (quart.), 48.1
(CH2(1Ј)), 39.2 (CH2(4Ј)), 26.3 (CH2), 24.3 (CH2). 19F NMR
8.72 Hz, JH,H = 6.95 Hz, 1H, H-C(7)), 7.66 (d, JH,H = 9.34
3
Hz, 1H, H-C(3)), 7.45 (d, JH,H = 9.34 Hz, 1H, H-C(4)), 7.37
3
3
(d, JH,H = 8.72 Hz, 1H, H-C(5)), 7.33 (dd, JH,H = 8.72 Hz,
3JH,H = 6.95 Hz, 1H, H-C(6)), 3.77Ϫ3.72 (m, 2H, CH2(1Ј)),
2.87Ϫ2.82 (m, 2H, CH2(7Ј)), 1.74Ϫ1.66 (m, 2H, CH2(2Ј)),
1.55Ϫ1.48 (m, 2H, CH2(6Ј)), 1.30Ϫ1.22 (m, 6H,
3 ϫ
CH2(3ЈϪ5Ј)). 13C NMR (100 MHz, D2O): δ = 156.3 (quart.),
135.6 (CH), 129.1 (CH), 120.4 (CH), 120.2 (q, 1JC,F = 257
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

576 Csuk et al.
Arch. Pharm. Pharm. Med. Chem. 2004, 337, 571−578
Hz, OCF3), 118.0 (CH), 48.6 (CH2(1Ј)), 39.6 (CH2(7Ј)), 29.0
(CH2), 28.0 (CH2), 27.8 (CH2), 25.9 (CH2), 25.7 (CH2). 19F
NMR (188 MHz, D2O): δ = Ϫ58.9 (OCF3). MS (ESI, 4.1 kV,
8 µL/min, N2, methanol): 392 (100%) [MH]+.
2-((2-{[3-(9-{2-Trifluoromethoxyacridinyl}amino)propyl]-
amino}-2-oxoethyl){2-[(2-{[3-(9-{2-trifluoromethoxyacridinyl}-
amino)propyl] amino}-2-oxoethyl)(carboxymethyl)amino]-
ethyl}amino) acetic acid (24)
As described for 23, from 17 (0.5 g, 1.23 mmol) and EDTA
dianhydride (141 mg, 0.55 mmol), 24 (220 mg, 43%) was
obtained as an orange, amorphous solid. UV-vis (methanol):
λmax (log ε) = 280 nm (4.68). IR (KBr): ν = 3281s, 1652s,
N1-[9-(2-Trifluoromethoxyacridinyl)]-octane-1,8-diamine bis-
hydrochloride (22)
1574s, 1404m, 1258s, 1117m cmϪ1 1H NMR (500 MHz,
.
DMSO-D6/D2SO4): δ = 8.50Ϫ8.42 (m, 2H, 2 ϫ H-C(8)), 8.43
(s, 2H, 2 ϫ H-C(1)), 8.04Ϫ7.93 (m, 6H, 6 ϫ H-C(3,4,7)), 7.85
(d, JH,H = 8.92 Hz, 2H, 2 ϫ H-C(5)), 7.55Ϫ7.49 (m, 2H, 2 ϫ
H-C(6)), 4.12Ϫ4.06 (m, 4H, 2 ϫ CH2(1Ј)), 3.91 (s, 4H, 2 ϫ
CH2(2Љ)), 3.81 (s, 4H, 2 ϫ CH2(3Љ)), 3.32 (s, 4H, 2 ϫ CH2(1Љ)),
3.26Ϫ3.21 (m, 4H, 2 ϫ CH2(3Ј)), 2.08Ϫ2.02 (m, 4H, 2 ϫ
CH2(2Ј)). 13C NMR (125 MHz, DMSO-D6/D2SO4): δ = 172.3
(C=O), 169.8 (C=O), 169.3 (quart.), 166.8 (quart.), 157.6
As described for 9, from 30 (2.99 g, 5.9 mmol), 22 (2.6 g,
92%) was obtained as a yellow, amorphous solid. UV-vis
(methanol): λmax (log ε) = 285 nm (4.56). IR (KBr): ν = 3424m,
2928s, 1629m, 1597s, 1540m, 1487s, 1366w, 1256s, 1212s,
3
1162s cmϪ1. 1H NMR (400 MHz, CD3OD): δ = 8.53 (d, 3JH,H
=
=
3
8.72 Hz, 1H, H-C(8)), 8.47 (s, 1H, H-C(1)), 7.99 (dd, JH,H
3
3
8.72 Hz, JH,H = 6.95 Hz, 1H, H-C(7)), 7.95 (d, JH,H = 9.34
Hz, 1H, H-C(4)), 7.92 (dd, JH,H = 9.34 Hz, JH,H = 2.28 Hz,
3
4
(quart.), 157.5 (quart.), 135.6 (quart.), 138.2 (quart.), 129.0
3
1H, H-C(3)), 7.85 (d, JH,H = 8.72 Hz, 1H, H-C(5)), 7.59 (dd,
1
(CH), 123.9 (CH), 121.4 (CH), 121.3 (CH), 120.2 (q, JC,F
=
3JH,H = 8.72 Hz, JH,H = 6.95 Hz, 1H, H-C(6)), 4.17Ϫ4.12 (m,
3
258 Hz, OCF3), 118.8 (CH), 118.7 (CH), 55.4 (CH2(3Љ)), 54.4
(CH2(2Љ)), 50.7 (CH2(1Љ)), 46.7 (CH2(1Ј)), 36.3 (CH2(3Ј)), 28.7
(CH2(2Ј)). 19F NMR (188 MHz, DMSO-D6): δ = Ϫ57.7 (OCF3).
MS (ESI, 4.1 kV, 8 µL/min, N2, methanol): 483 (100%)
[MHK]2+, 927 (10%) [MH]+, 965 (50%) [MK]+. Analysis for
C44H44F6N8O8 (926.88): C 57.02; H 4.79; N 12.09; found: C
56.91; H 4.93; N 11.88.
2H, CH2(1Ј)), 2.93Ϫ2.88 (m, 2H, CH2(8Ј)), 2.05Ϫ1.97 (m, 2H,
CH2(2Ј)), 1.70Ϫ1.62 (m, 2H, CH2(7Ј)), 1.54Ϫ1.38 (m, 6H,
4 ϫ CH2(3ЈϪ6Ј)). 13C NMR (100 MHz, CD3OD): δ = 159.2
(quart.), 136.7 (CH), 130.2 (CH), 125.2 (CH), 122.0 (CH),
121.9 (q, 1JC,F = 257 Hz, OCF3), 119.6 (CH), 50.5 (CH2(1Ј)),
40.8 (CH2(8Ј)), 30.5 (CH2), 30.0 (2 CH2), 28.5 (CH2), 27.7
(CH2), 27.4 (CH2). 19F NMR (188 MHz, CD3OD): δ = Ϫ60.4
(OCF3). MS (ESI, 4.1 kV, 8 µL/min, N2, methanol): 406
(100%) [MH]+.
2-((2-{[4-(9-{2-Trifluoromethoxyacridinyl}amino)butyl]amino}-
2-oxoethyl){2-[(2-{[4-(9-{2-trifluoromethoxyacridinyl}-
amino)butyl]amino}-2-oxoethyl)(carboxymethyl)amino]-
ethyl}amino) acetic acid (25)
2-((2-{[2-(9-{2-Trifluoromethoxyacridinyl}amino)ethyl]amino}-
2-oxoethyl){2-[(2-{[2-(9-{2-trifluoromethoxyacridinyl}amino)-
ethyl]amino}-2-oxoethyl)(carboxymethyl)amino]ethyl}amino)
acetic acid (23)
As described for 23, from 18 (0.48 g, 1.14 mmol) and EDTA
dianhydride (131 mg, 0.51 mmol), 25 (60 mg, 12%) was ob-
tained as an orange, amorphous solid. UV-vis (methanol):
λmax (log ε) = 281 nm (4.99). IR (KBr): ν = 3261m, 2929m,
1652s, 1566s, 1478m, 1395m, 1256s, 1217s, 1160s cmϪ1
.
A solution of 16 (304 mg, 0.77 mmol) in dry DMF (10 mL)
containing triethylamine (3 mL) was stirred at room tempera-
ture for 10 min; EDTA dianhydride (99 mg, 0.39 mmol) was
added, and stirring was continued at room temperature for
another 12 h. After addition of acetone (10 mL) and hexane
(40 mL), the crude product was filtered off and dissolved in
methanol (15 mL) containing aq. hydrochloric acid (10%, 0.5
mL). The solvents were removed in vacuo, and the product
was purified by chromatography (silica gel, methanol/aceto-
nitrile/ammonia 70 : 25 : 5) to afford 23 (80 mg, 23%) as an
amorphous solid. UV-vis (methanol): λmax (log ε) = 280 nm
(4.91). IR (KBr): ν = 3266m, 1590s, 1479m, 1435m, 1403m,
1H NMR (500 MHz, DMSO-D6/D2SO4): δ = 8.50Ϫ8.45 (m,
2H, 2 H-C(8)), 8.05Ϫ7.90 (m, 8H, 8 ϫ H-C(1,3,4,7)), 7.87
3
(d, JH,H = 8.73 Hz, 2H, 2 ϫ H-C(5)), 7.55Ϫ7.50 (m, 2H, 2 ϫ
H-C(6)), 4.09Ϫ4.04 (m, 4H, 2 ϫ CH2(1Ј)), 4.07 (s, 4H, 2 ϫ
CH2(2Љ)), 3.95 (s, 4H, 2 ϫ CH2(3Љ)), 3.55 (s, 4H, 2 ϫ CH2(1Љ)),
3.16Ϫ3.11 (m, 4H, 2 ϫ CH2(4Ј)), 1.92Ϫ1.85 (m, 8H, 4 ϫ
CH2(2Ј,3Ј)), 1.56Ϫ1.50 (m, 8H, 4 ϫ CH2(2Ј,3Ј)). 13C NMR
(125 MHz, DMSO-D6/D2SO4): δ = 170.6 (C=O), 168.2 (C=O),
165.1 (quart.), 158.1 (quart.), 157.6 (quart.), 143.8 (quart.),
139.5 (quart.), 138.2 (quart.), 129.5 (CH), 124.9 (CH), 124.8
1
(CH), 121.9 (CH), 121.5 (CH), 120.3 (q, JC,F = 257 Hz,
1332m, 1255s, 1216s, 1160s cmϪ1 1H NMR (500 MHz,
.
OCF3), 119.2 (CH), 118.9 (CH), 111.8 (quart.), 111.6 (quart.),
55.5 (CH2(3Љ)), 54.3 (CH2(2Љ)), 50.3 (CH2(1Љ)), 48.6 (CH2(1Ј)),
38.5 (CH2(4Ј)), 26.3 (CH2), 26.1 (CH2). 19F NMR (188 MHz,
DMSO-D6): δ = Ϫ57.7 (OCF3). MS (ESI, 4.1 kV, 8 µL/min,
N2, methanol): 497 (100%) [MHK]2+, 955 (4%) [MH]+, 993
(15%) [MK]+. Analysis for C46H48F6N8O8 (954.93): C 57.86;
H 5.07; H 11.73; found: C 57.62; H 5.23; N 11.63.
DMSO-D6/D2SO4): δ = 8.51 (s, 2H, 2 ϫ H-C(1)), 8.50 (d,
3JH,H = 8.64 Hz, 2H, 2 ϫ H-C(8)), 8.00Ϫ7.95 (m, 6H, 6 ϫ H-
3
C(3,4,7)), 7.83 (d, JH,H = 8.64 Hz, 2H, 2 ϫ H-C(5)),
7.53Ϫ7.49 (m, 2H, 2
H-C(6)), 4.16Ϫ4.13 (m, 4H, 2 ϫ
CH2(1Ј)), 3.87 (s, 4H, 2 ϫ CH2(2Љ)), 3.79 (s, 4H, 2 ϫ CH2(3Љ)),
3.66Ϫ3.61 (m, 4H, 2 ϫ CH2(2Ј)), 3.25 (s, 4H, 2 ϫ CH2(1Љ)).
13C NMR (125 MHz, DMSO-D6/D2SO4): δ = 169.7 (C=O),
168.48 (C=O), 168.44 (quart.), 157.5 (quart.), 157.4 (quart.),
143.7 (quart.), 139.5 (quart.), 138.2 (quart.), 135.7 (CH),
129.0 (CH), 124.6 (CH), 121.44 (CH), 121.35 (CH), 120.2 (q,
1JC,F = 257 Hz, OCF3), 118.8 (CH), 118.7 (CH), 109.7 (quart.),
55.3 (CH2(3Љ)), 54.2 (CH2(2Љ)), 50.8 (CH2(1Љ)), 48.6 (CH2(1Ј)),
38.2 (CH2(2Ј)). 19F NMR (188 MHz, DMSO-D6): δ = Ϫ57.7
(OCF3). MS (ESI, 4.1 kV, 8 µL/min, N2, methanol): 469
(100%) [MHK]2+, 899 (14%) [MH]+, 837 (27%) [MK]+. Analy-
sis for C42H40F6N8O8 (898.83): C 56.13; H 4.49; N 12.47;
found: C 55.97; H 4.61; N 12.32.
2-((2-{[5-(9-{2-Trifluoromethoxyacridinyl}amino)pentyl]-
amino}-2-oxoethyl){2-[(2-{[5-(9-{2-trifluoromethoxyacridinyl}
amino)pentyl]amino}-2-oxoethyl)(carboxymethyl)amino]-
ethyl}amino) acetic acid (26)
As described for 23, from 19 (0.5 g, 1.15 mmol) and EDTA
dianhydride (132 mg, 0.52 mmol), 26 (80 mg, 16%) was ob-
tained as an orange, amorphous solid. UV-vis (methanol):
λmax (log ε) = 284 nm (5.01). IR (KBr): ν = 3406m, 2933m,
1565s, 1523m, 1478m, 1436m, 1397m, 1330w, 1257s,
© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Arch. Pharm. Pharm. Med. Chem. 2004, 337, 571−578
Pathogen-inactivating Intercalators 577
1
1217s, 1160m cmϪ1. H NMR (500 MHz, DMSO-D6/D2SO4):
(C=O), 157.4 (quart.), 129.3 (2 ϫ CH), 121.0 (2 ϫ CH), 120.0
(CH), 119.15 (2 ϫ CH), 56.0 (CH2(3Љ)), 55.0 (CH2(2Љ)), 50.5
(CH2(1Љ)), 49.0 (CH2(1Ј)), 40.3 (CH2(7Ј)), 28.7 (CH2), 28.4 (2
ϫ CH2), 26.2 (2 ϫ CH2). 19F NMR (188 MHz, CD3OD/TFA):
δ = Ϫ78.3 (OCF3). MS (ESI, 4.1 kV, 8 µL/min, N2, methanol):
δ = 8.69Ϫ8.40 (br m, 4H, 4 ϫ H-C(1,8)), 8.01Ϫ7.95 (m, 6H,
6 ϫ H-C(3,4,7)), 7.87 (d, JH,H = 8.94 Hz, 2H, 2 ϫ H-C(5)),
3
7.56Ϫ7.51 (m, 2H, 2 ϫ H-C(6)), 4.09 (s, 4H, 2 ϫ CH2(2Љ)),
4.06Ϫ4.01 (m, 4H, 2 ϫ CH2(1Ј)), 3.97 (s, 4H, 2 ϫ CH2(3Љ)),
3.56 (s, 4H, 2 ϫ CH2(1Љ)), 3.13Ϫ3.08 (m, 4H, 2 ϫ CH2(5Ј)),
1.91Ϫ1.84 (m, 4H, 2 ϫ CH2(2Ј)), 1.51Ϫ1.43 (m, 4H, 2 ϫ
CH2(4Ј)), 1.42Ϫ1.34 (m, 4H, 2 ϫ CH2(3Ј)). 13C NMR (125
MHz, DMSO-D6/D2SO4): δ = 169.1 (C=O), 168.2 (C=O),
165.0 (quart.), 157.6 (quart.), 157.5 (quart.), 143.8 (quart.),
139.5 (quart.), 135.7 (CH), 129.1 (CH), 124.8 (CH), 121.53
539 (40%) [MHK]2+
,
1077 (100%) [MK]+. Analysis for
C52H60F6N8O8 (1039.10): C 60.11; H 5.82; N 10.78; found: C
59.96; H 5.95; N 10.61.
2-((2-{[8-(9-{2-Trifluoromethoxyacridinyl}amino)octyl]amino}-
2-oxoethyl){2-[(2-{[8-(9-{2-trifluoromethoxyacridinyl}amino)-
octyl]amino}-2-oxoethyl)(carboxymethyl)amino]ethyl}amino)
acetic acid (29)
1
(CH), 121.43 (CH), 120.4 (q, JC,F = 257 Hz, OCF3), 118.96
(CH), 118.85 (CH), 111.8 (quart.), 111.6 (quart.), 55.5
(CH2(3Љ)), 54.3 (CH2(2Љ)), 50.3 (CH2(1Љ)), 49.0 (CH2(1Ј)), 38.7
(CH2(5Ј)), 28.5 (2 ϫ CH2), 23.7 (CH2). 19F NMR (188 MHz,
DMSO-D6): δ = Ϫ57.7 (OCF3). MS (ESI, 4.1 kV, 8 µL/min,
N2, methanol): 511 (100%) [MHK]2+, 1021 (9%) [MK]+. Analy-
sis for C48H52F6N8O8 (982.99): C 58.65; H 5.33; N 11.40;
found: C 58.51; H 5.56; N 11.21.
As described for 23, from 22 (0.5 g, 1.05 mmol) and EDTA
dianhydride (120 mg, 0.47 mmol), 29 (62 mg, 12%) was ob-
tained as an orange, amorphous solid. UV-vis (methanol):
λmax (log ε) = 280 nm (4.93). IR (KBr): ν = 3260m, 3077m,
2930s, 2856m, 1648s, 1592s, 1478m, 1399m, 1362m,
1330w, 1258s, 1216s, 1162s cmϪ1 1H NMR (500 MHz,
.
DMSO-D6/D2SO4): δ = 8.51Ϫ8.40 (br m, 4H, 4 ϫ H-C(1,8)),
7.99Ϫ7.91 (m, 6H, 6 ϫ H-C(3,4,7)), 7.85 (d, JH,H = 8.46 Hz,
2H, 2 ϫ H-C(5)), 7.53Ϫ7.47 (m, 2H, 2 ϫ H-C(6)), 4.10 (s,
4H, 2 ϫ CH2(2Љ)), 4.03Ϫ3.95 (m, 4H, 2 ϫ CH2(1Ј)), 3.97 (s,
4H, 2 ϫ CH2(3Љ)), 3.59 (s, 4H, 2 ϫ CH2(1Љ)), 3.06Ϫ3.00 (m,
2-((2-{[6-(9-{2-Trifluoromethoxyacridinyl}amino)hexyl]amino}-
2-oxoethyl){2-[(2-{[6-(9-{2-trifluoromethoxyacridinyl}amino)-
hexyl]amino}-2-oxoethyl)(carboxymethyl)amino]ethyl}amino)
acetic acid (27)
3
4H,
2 ϫ CH2(8Ј)), 1.86Ϫ1.77 (m, 4H, 2 ϫ CH2(2Ј)),
As described for 23, from 20 (0.5 g, 1.11 mmol) and EDTA
dianhydride (128 mg, 0.5 mmol), 27 (122 mg, 24%) was ob-
tained as an orange, amorphous solid. UV-vis (methanol):
λmax (log ε) = 283 nm (4.92). IR (KBr): ν = 3423s, 2935m,
1647s, 1593s, 1485m, 1401m, 1364m, 1259s, 1208s, 1163s
1.38Ϫ1.15 (m, 20H, 10 ϫ CH2(3ЈϪ7Ј)). 13C NMR (100 MHz,
DMSO-D6/D2SO4): δ = 169.7 (C=O), 166.8 (C=O), 157.2 (2
ϫ quart.), 146.3 (quart.), 135.4 (CH), 128.7 (CH), 125.1 (CH),
1
121.1 (CH), 121.2 (CH), 120.6 (q, JC,F = 257 Hz, OCF3),
1
118.6 (CH), 118.5 (CH), 112.6 (2 ϫ quart.), 55.6 (CH2(3Љ)),
54.1 (CH2(2Љ)), 51.0 (CH2(1Љ)), 48.9 (CH2(1Ј)), 38.7 (CH2(6Ј)),
28.8 (CH2), 28.7 (CH2), 28.6 (2 ϫ CH2), 26.3 (CH2), 26.1
(CH2). 19F NMR (188 MHz, DMSO-D6): δ = Ϫ57.8 (OCF3).
MS (ESI, 4.1 kV, 8 µlL/min, N2, methanol): 553 (100%)
[MHK]2+, 1065 (18%) [MK]+. Analysis for C54H64F6N8O8
(1067.15): C 60.78; H 6.05; N 10.50; found: C 60.61; H 6.23;
N 10.43.
cmϪ1. H NMR (500 MHz, DMSO-D6/D2SO4): δ = 8.55Ϫ8.37
(br m, 4H, 4 ϫ H-C(1,8)), 8.02Ϫ7.89 (m, 6H, 6 ϫ H-C(3,4,7)),
7.85 (d, 3JH,H = 8.57 Hz, 2H, 2 ϫ H-C(5)), 7.52Ϫ7.47 (m, 2H,
2 ϫ H-C(6)), 4.11 (s, 4H, 2 ϫ CH2(2Љ)), 4.04Ϫ3.96 (m, 4H, 2
ϫ CH2(1Ј)), 3.99 (s, 4H, 2 ϫ CH2(3Љ)), 3.62 (s, 4H, 2 ϫ
CH2(1Љ)), 3.07Ϫ3.01 (m, 4H, 2 ϫ CH2(6Ј)), 1.86Ϫ1.79 (m,
4H, 2 ϫ CH2(2Ј)), 1.42Ϫ1.23 (m, 12H, 6 ϫ CH2(3ЈϪ5Ј)). 13C
NMR (125 MHz, DMSO-D6/D2SO4): δ = 168.0 (C=O), 164.7
(C=O), 158.2 (quart.), 157.6 (quart.), 144.0 (quart.), 139.7
(quart.), 138.4 (quart.), 135.9 (CH), 129.3 (CH), 125.1 (CH),
1
121.73 (CH), 121.63 (CH), 120.6 (q, JC,F = 257 Hz, OCF3),
References
119.15 (CH), 119.05 (CH), 112.0 (quart.), 111.8 (quart.), 55.7
(CH2(3Љ)), 54.6 (CH2(2Љ)), 50.4 (CH2(1Љ)), 49.1 (CH2(1Ј)), 39.0
(CH2(6Ј)), 28.9 (2 ϫ CH2), 26.31 (CH2), 26.24 (CH2). 19F
NMR (188 MHz, DMSO-D6): δ = Ϫ57.8 (OCF3). MS (ESI, 4.1
kV, 8 µL/min, N2, methanol): 525 (100%) [MHK]2+, 1049
(20%) [MK]+. Analysis for C50H56F6N8O8 (1011.04): C 59.40;
H 5.59; N 11.08; found: C 59.32; H 5.78; N 10.94.
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[2] R. Csuk, A. Barthel, C. Raschke, Tetrahedron 2004,
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2-((2-{[7-(9-{2-Trifluoromethoxyacridinyl}amino)heptyl]-
amino}-2-oxoethyl){2-[(2-{[7-(9-{2-trifluoromethoxyacridinyl}-
amino)heptyl amino}-2-oxoethyl)(carboxymethyl)amino]-
ethyl}amino) acetic acid (28)
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As described for 23, from 21 (0.5 g, 1.08 mmol) and EDTA
dianhydride (124 mg, 0.48 mmol), 28 (63 mg, 13%) was ob-
tained as an orange, amorphous solid. UV-vis (methanol):
λmax (log ε) = 281 nm (5.03). IR (KBr): ν = 3323s, 2936m,
1650s, 1593s, 1488m, 1399m, 1364m, 1260s, 1211s, 1160s
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1
cmϪ1. H NMR (500 MHz, DMSO-D6/D2SO4): δ = 8.57Ϫ8.38
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(br m, 4H, 4 ϫ H-C(1,8)), 8.07Ϫ7.95 (m, 6H, 6 ϫ H-C(3,4,7)),
7.87 (d, 3JH,H = 8.75 Hz, 2H, 2 ϫ H-C(5)), 7.57Ϫ7.51 (m, 2H,
2 ϫ H-C(6)), 4.09Ϫ4.02 (m, 4H, 2 ϫ CH2(1Ј)), 4.07 (s, 4H, 2
ϫ CH2(2Љ)), 3.94 (s, 4H, 2 ϫ CH2(3Љ)), 3.53 (s, 4H, 2 ϫ
CH2(1Љ)), 3.09Ϫ3.03 (m, 4H, 2 ϫ CH2(7Ј)), 1.89Ϫ1.82 (m,
4H, 2 ϫ CH2(2Ј)), 1.42Ϫ1.18 (m, 16H, 8 ϫ CH2(3ЈϪ6Ј)). 13C
NMR (125 MHz, DMSO-D6/D2SO4): δ = 168.2 (C=O), 164.7
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T H E N E W S T A N D A R D
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