Synthesis of dimeric acridine derived nucleic acid intercalators

Article abstract of DOI:10.1515/znb-2005-0114

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. Suitably monoprotected 1,ω-alkyldiamines gave upon reaction with 6,9-dichloro-2-methoxyacridine (1) followed by deprotection and reaction with EDTA dianhydride the target molecules. In the presence of ascorbate a reduction of the phage-titer of the MS2 phages by > 8 logarithmic decades was achieved.

Full text of DOI:10.1515/znb-2005-0114

Synthesis of Dimeric Acridine Derived Nucleic Acid Intercalators
Ren e´ Csuk, Thorsten Brezesinski, Gunnar G o¨ the, Christian Raschke, and Stefan Reißmann
Institut f u¨ r Organische Chemie, Martin-Luther-Universit a¨ t Halle-Wittenberg,
Kurt-Mothes-Str. 2, D-06120 Halle (Saale), Germany
Reprint requests to Prof. Dr. R. Csuk. E-mail: csuk@chemie.uni-halle.de
Z. Naturforsch. 60b, 89 – 98 (2005); received June 28, 2004
A series of antiviral compounds consisting of an intercalating acridine derived part, a spacer re-
gion and a reactive EDTA-derived conjugate was synthesized in an easy sequence. Suitably mono-
protected 1,ω-alkyldiamines gave upon reaction with 6,9-dichloro-2-methoxyacridine (1) followed
by deprotection and reaction with EDTA dianhydride the target molecules. In the presence of ascor-
bate a reduction of the phage-titer of the MS2 phages by > 8 logarithmic decades was achieved.
Key words: Acridine, Antivirals, Intercalators, Fenton-Mechanism
Introduction
Blood is a critical element of medical treatment. Ev-
ery day, numerous units of blood are transfused for a
variety of illnesses and conditions including accidents,
burns, heart surgery, organ transplants, leukemia, can-
cer, sickle cell anemia, thalassemia, hemophilia, and
others.
Although the blood supply is currently much safer
than it has been in the past due to various efforts includ-
ing increased testing and more stringent donor screen-
ing criteria, blood transfusions, unfortunately them-
selves can be a cause of illness. However, pathogens –
viruses, bacteria, and parasites – are still transmit-
ted from person-to-person through donated and trans-
fused blood. The testing and donor screening that
are used in blood collection facilities are only able
to identify known pathogens. Additionally, new vari-
ants and strains of existing infectious agents continue
to emerge, moving beyond the sensitivity of existing
tests.
Fig. 1. Basic structures of
the pathogen inactivating
compounds.
pathogens combined with a conjugate that destroys the
nucleic acid subsequently via a Fenton [2, 3] mecha-
nism. The conjugate consisted of a metal-chelate com-
plex wherein the metal can change between at least
two levels of oxidation. These compounds preferen-
tially showed an acridine as the intercalator possess-
ing in addition a spacer group to an EDTA-Fe(II/III)
chelate [4, 5] acting as the conjugate. Damage to the
DNA was accomplished by producing OH radicals by
a Fe(II) catalyzed Fenton mechanism [1, 6, 7]. Further-
more, the addition of a reducing agent such as ascor-
bate [8 – 11] leads to a cycle which increases the dam-
age to the biological molecules.
A second issue with testing is the presence of a
window period” in which an individual is infected,
Results and Discussion
”
but the virus is not present in large enough quantities
From preliminary modelling studies we concluded
to be detected. However, these quantities are enough that an improved antipathogenic performance could be
to infect a patient with blood. Although improved test- expected from compounds exhibiting a stronger abil-
ing has decreased the window period e.g. for HIV from ity to interact with DNA or RNA. Thus, from a bet-
2
0 days to 11 days, even at this level, however, tainted ter intercalating action of the compounds a higher de-
blood can slip through.
gree of viral inactivation should be expected. Inspec-
Quite recently, a new type of pathogen inactivating tion of molecule models indicated that the length of
agents [1] (Fig. 1, type A) was introduced – consist- the spacer between the aromatic rest and the chelator
ing of an intercalator that binds to the nucleic acid of should be of some significance with respect to aspects
0
932–0776 / 05 / 0100–0089 $ 06.00 ꢀc 2005 Verlag der Zeitschrift f u¨ r Naturforschung, T u¨ bingen · http://znaturforsch.com
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Ren e´ Csuk et al. · Synthesis of Dimeric Acridine Derived Nucleic Acid Intercalators
Fig. 2. Inactivation of phage MS2 as a function of spacer
length (5 mmol Na-ascorbate, 101 µmol of compounds, 3
3
+
equiv. Fe loaded, incubation time 4 h) and of the tempera-
◦
◦
ture (−25 C, −36 C).
Scheme 1. a) (Boc) O, dioxane; b) phenol; c) aq. HCl;
d) NEt3.
2
For biological screening, the well established sys-
tem containing MS2 bacteriophages (genus of the fam-
ily leviviridae; uncoated, containing ssRNA) was used;
these viruses contain the short version of the genome
and have a separate gene for cell lysis.
of binding and interaction. From these modelling stud-
ies we also concluded, that the use of substituted acri-
dine moieties should be advantageous for an improved
intercalation. Therefore, the synthesis of dimeric struc-
tures (Fig. 1, type B) was planned using 6-chloro-2-
methoxy-acridines for the construction of the interca-
lating moiety.
As far as the synthesis (Scheme 1) of this spacer
is concerned, a variety of 1,ω-diamines was mono-
protected by the reaction with tert-butyl-dicarbonate
in dry dioxane to afford the corresponding 1-amino-
x-(tert-butyloxycarbonyl)amino-alkanes2 [12], 3 [13],
In this preliminary screening, several of the target
compounds were treated with an 2 – 5 molar excess
3+
of Fe , lyophilized and incubated with the phages in
Tris-buffer in the presence of sodium ascorbate. A re-
duction of the phagetiter of the MS2 phages by > 8
logarithmic decades was achieved.
From these experiments it can be assumed, that the
inactivation of the virus depends both on the tempera-
ture, the time of incubation as well as on the concen-
tration of the added ascorbate. Increased activity with
increased concentration of ascorbate as well as the ob-
servation that no activity is associated with these com-
pounds in the lack of ascorbate allows a triggering of
the activity by the addition of ascorbic acid.
4
1
[14], 5 [15], 6 [16], 7 [17], 8 [18], 9 [19], 10 [18],
1 [20]. Thus, usually two equivalents of the diamine
were allowed to react with one equivalent of tert-butyl-
dicarbonate to result in approx. 40% yields of the cor-
responding products. These moderate yields could be
improved, however, by using a 5-6 molar excess of the
amine.
Modifications in the structure of these compounds
as well an extended biological screening are presently
performed in our laboratories.
These 1-amino-x-(tert-butyloxycarbonyl)amino-al-
kanes 2 – 11 were allowed to react with 6,9-dichloro-
Experimental Section
2
2
-methoxyacridine (1) in the presence of phenol [21–
4] to result in the formation of the corresponding tert-
General
butyl N-[x-(9-acridinyl-amino)alkyl]-carbamates 12 –
The melting points are uncorrected (Reichert hot stage
microscope), NMR spectra (internal Me4Si) were recorded
using either a Bruker AM250 or a Varian XL300 instrument
2
1. Deprotection of these compounds by aq. hydro-
◦
chloric acid for 10 h at 30 C gave the corresponding
bishydrochlorides 22– 31. Reaction of these amines
with EDTA dianhydride finally furnished the target Perkin-Elmer 298 instrument, MS spectra were taken either
(δ given in ppm, J in Hz), IR spectra (film or KBr pellet) on a
compounds 32-40.
on a MAT311A or a Varian-112S instrument; for elemen-
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91
tal analysis a Foss-Heraeus Vario EL instrument was used. tert-Butyl N-(4-aminobutyl)carbamate (4)
TLC was performed on silica gel (Merck 5554, detection by
Following GP1 from 1,4-diaminobutane (10.0 g,
dipping in a solution containing 10% sulfuric acid (400 ml),
ammonium molybdate (20 g) and cerium(IV) sulfate (20 mg)
1
13.0 mmol) 4 (4.2 g, 39%) was obtained as a viscous
1
oil. H NMR (400 MHz, CDCl ): δ = 4.68 (br s, 1 H,
◦
3
followed by heating to 150 C. All reactions were performed
NH), 3.08 – 3.06 (m, 2 H, CH (4)), 2.68 – 2.65 (m, 2 H,
2
under dry argon.
CH (1)), 1.50 – 1.42 (m, 4 H, 2 × CH (2,3), 1.39 (s, 9 H,
2
2
1
3
tBu). – C NMR (100 MHz, CDCl ): δ = 155.8 (CO),
8.9 (tBu), 41.8 (CH ), 40.5 (CH ), 30.9 (CH ), 28.5 (tBu),
3
General procedure for the synthesis of 1-amino-x-(tert-butyl-
oxycarbonyl)amino-alkanes (GP1)
7
2
2 2 2
7.6 (CH ). – MS (ESI, 4.1 kV, 8 µl/min, N , methanol):
2
2
i
+
+
To a suspension of the corresponding alkyl-1,x-diamine in m/z = 133 (8%) [(M- butene)H] , 189 (100%) [MH] . –
◦
dry dioxane (200 ml) at 0 C within 5 h a solution of di-tert- Analysis for C H N O (188.27): C 57.42, H 10.71,
9
20
2 2
butyl-dicarbonate (18.1 g, 83 mmol) in dry dioxane (200 ml) N 14.88; found C 57.29, H 10.94, N 14.74.
was slowly added. After stirring for an additional 6 h, the
solvent was removed under reduced pressure and the residue
extracted with ether (3 × 150 ml); the ether phase was dis-
carded. To the combined extracts water (200 ml) was added,
the pH-value of the aqueous phase adjusted to 3 – 4, the aq.
tert-Butyl N-(5-aminopentyl)carbamate (5)
Following GP1 from 1,5-diaminopentane (15.3 g,
0
.15 mol) 5 (6.0 g, 39.5%) was obtained as a slightly yel-
1
lowish viscous oil. H NMR (400 MHz, CDCl ): δ = 4.53
3
layer was quickly separated and an aq. solution of Na CO
2
3
(
(
(
(
(
(
2
br s, 1 H, NH), 3.09 – 3.06 (m, 2 H, CH (5)), 2.67 – 2.64
2
(
saturated, 100 ml) was added. The aq. phase was extracted
with chloroform (3 × 150 ml), the combined extracts were
dried (Na SO ), and the solvent was removed under reduced
m, 2 H, CH (1)), 1.50 – 1.42 (m, 4 H, 2 × CH (2,4), 1.41
2
2
1
3
s, 9 H, tBu), 1.39 – 1.31 (m, 2 H, CH (3)). – C NMR
2
2
4
50.3 MHz, CDCl ): δ = 156.0 (CO), 79.1 (tBu), 42.1
CH ), 40.5 (CH ), 30.4 (CH ), 29.9 (CH ), 28.4 (tBu), 24.1
CH ). – MS (ESI, 4.1 kV, 8 µl/min, N , methanol): m/z =
03 (100%) [MH] . – Analysis for C H N O (202.29):
3
pressure to afford the crude products that were used for the
next steps without any further purification. Analytically pure
samples, however, were obtained after chromatography (RP-
2
2
2
2
2
2
+
1
0 22 2 2
1
3
8, methanol/water 9:1). Compounds 12 – 21, 22 – 31 and
2 – 41 are members of homologous series and thus similar in
C 59.37, H 10.96, N 13.85; found C 59.22, H 10.64, N 13.97.
their spectra; therefore, only representative values are given.
tert-Butyl N-(6-aminohexyl)carbamate (6)
tert-Butyl N-(2-aminoethyl)carbamate (2)
Following GP1 from 1,6-diamino-hexane (19.0 g,
1
63 mmol) 6 (7.3 g, 40%) was obtained as a viscous oil.
Following GP1 from 1,2-diaminoethane (10.0 g,
1
H NMR (400 MHz, CDCl ): δ = 4.62 (br s, 1 H, NH),
3
0
.17 mol) 2 (6.3 g, 47.4%) was obtained as a viscous
3
1
.05 – 3.00 (m, 2 H, CH (6)), 2.61 – 2.58 (m, 2 H, CH (1)),
oil. H NMR (400 MHz, CDCl ): δ = 4.90 (br s, 1 H,
2
2
3
1
1
.42 – 1.38 (m, 4 H, 2×CH (2,5), 1.36 (s, 9 H, tBu), 1.26 –
NH), 3.15 – 3.11 (m, 2 H, CH (2)), 2.77 – 2.74 (m, 2 H,
2
2
13
13
.23 (m, 4 H, 2×CH (3,4)). – C NMR (100 MHz, CDCl ):
CH (1)), 1.41 (s, 9 H, tBu). – C NMR (100 MHz, CDCl3):
2
3
2
δ = 155.8 (CO), 78.8 (tBu), 42.1 (CH ), 40.5 (CH ), 33.7
δ = 156.1 (CO), 79.1 (tBu), 41.9 (CH ), 40.8 (CH ),
2
2
2
2
(CH ), 30.0 (CH ), 28.4 (tBu), 26.6 (CH ), 26.5 (CH ). –
2
8.4 (tBu). – MS (ESI, 4.1 kV, 8 µl/min, N , methanol):
2 2 2 2
2
i
+
+
MS (ESI, 4.1 kV, 8 µl/min, N , methanol): m/z = 161
m/z = 105 (7%) [(M- butene)H] , 161 (100%) [MH] . –
Analysis for C H N O (160.21): C 52.48, H 10.07,
2
i
+
+
(8%) [(M- butene)H] , 217 (100%) [MH] . – Analysis for
7
16
2 2
C H N O (216.32): C 51.07, H 11.18, N 12.85; found
C 50.85, H 11.46, N 12.67.
N 17.48; found C 52.37, H 10.23, N 17.58.
11 24
2
2
tert-Butyl N-(3-aminopropyl)carbamate (3)
tert-Butyl N-(7-aminoheptyl)carbamate (7)
Following GP1 from 1,3-diaminopropane (14.0 g,
0
.19 mol) 3 (8.5 g, 52%) was obtained as a slightly yellow-
Following GP1 from 1,7-diamino-heptane (15.0 g,
1
ish viscous oil. H NMR (400 MHz, CDCl ): δ = 4.97 (br 115 mmol) 7 (5.9 g, 44%) was obtained as a viscous
3
1
s, 1 H, NH), 3.17 – 3.12 (m, 2 H, CH (3)), 2.71 – 2.68 (m, oil. H NMR (400 MHz, CDCl ): δ = 4.49 (br s, 1 H,
2
3
2
H, CH (1)), 1.58 – 1.51 (m, 2 H, CH (2), 1.38 (s, 9 H, NH), 3.10 – 3.05 (m, 2 H, CH (7)), 2.67 – 2.63 (m, 2 H,
2
2
2
1
3
tBu). – C NMR (100 MHz, CDCl ): δ = 156.0 (CO), 79.0 CH (1)), 1.46 – 1.42 (m, 4 H, 2 × CH (2,6)), 1.42 (s, 9 H,
3
2
2
1
3
(
tBu), 39.8 (CH ), 38.5 (CH ), 33.6 (CH ), 28.5 (tBu). – tBu), 1.33 – 1.27 (m, 6 H, 3 × CH (3,4,5). – C NMR
2
2
2
2
MS (ESI, 4.1 kV, 8 µl/min, N , methanol): m/z = 119 (100 MHz, CDCl ): δ = 155.9 (CO), 78.9 (tBu), 42.2
2
3
i
+
+
(8%) [(M- butene)H] , 175 (100%) [MH] . – Analysis for (CH ), 40.6 (CH ), 33.8 (CH ), 30.0 (CH ), 28.4 (tBu),
2 2 2 2
C H N O (174.24): C 55.15, H 10.41, N 16.08; found 29.1 (CH ), 26.8 (CH ), 26.8 (CH ). – MS (ESI, 4.1 kV,
8
18
2
2
2
2
2
+
C 55.01, H 10.27, N 15.88.
8 µl/min, N , methanol): m/z = 131 (2%) [(M-Boc)H] ,
2
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1
2
Ren e´ Csuk et al. · Synthesis of Dimeric Acridine Derived Nucleic Acid Intercalators
i
+
+
75 (11%) [(M- butene)H] , 231 (100%) [MH] . – Anal- C H N O (272.43): C 66.13, H 11.84, N 10.28; found
1
5
32
2
2
ysis for C H N O (230.35): C 62.57, H 11.38, N 12.16; C 66.00, H 11.98, N 9.99.
1
26
2 2 2
found C 62.37, H 11.49, N 12.12.
tert-Butyl N-(12-aminododecyl)-carbamate (11)
tert-Butyl N-(8-aminooctyl)carbamate (8)
Following the procedure given for the synthesis of 10
from 1,12-diamino-dodecane (17.0 g, 85 mmol) and di-tert-
butyl-dicarbonate (9.3 g, 42.5 mmol) 11 (6.6 g, 52%) was
Following GP1 from 1,8-diaminooctane (15.7 g,
1
09 mmol) 8 (5.5 g, 41.3%) was obtained as a viscous oil.
1
1
obtained as a white amorphous solid. H NMR (400 MHz,
H NMR (400 MHz, CDCl ): δ = 4.53 (br s, 1 H, NH),
3
CDCl ): δ = 4.49 (br s, 1 H, NH), 3.08 – 3.06 (m, 2 H,
3
3
1
.09 – 3.04 (m, 2 H, CH (8)), 2.65 – 2.62 (m, 2 H, CH (1)),
2
2
CH (12)), 2.67 – 2.63 (m, 2 H, CH (1)), 1.42 – 1.37 (m, 4 H,
2
2
.40 – 1.35 (m, 4 H, 2 × CH (2,7)), 1.40 (s, 9 H, tBu), 1.26
2
1
3
2 × CH2(2.11)), 1.41 (s, 9 H, tBu), 1.29 – 1.24 (m, 16 H,
(
br s, 8H, 4 ×CH (3-6)). – C NMR (100 MHz, CDCl ):
2
3
13
8
(
×CH (3-10)). – C NMR (100 MHz, CDCl ): δ = 155.9
2
3
δ = 155.9 (CO), 78.9 (tBu), 42.3 (CH ), 40.7 (CH ), 33.9
2
2
CO), 78.9 (tBu), 42.3 (CH ), 40.6 (CH ), 33.9 (CH ),
2 2 2
(CH ), 30.2 (CH ), 29.5 (CH ), 29.3 (CH ), 28.5 (tBu),
2 2 2 2
3
0.1 (CH ), 29.7 (CH ), 29.6 (CH ), 29.6 (CH ), 29.5
2 2 2 2
2
6.9 (CH ), 26.8 (CH ). – MS (ESI, 4.1 kV, 8 µl/min,
2 2
+
(CH2), 29.5 (CH2), 29.3 (CH2), 28.5 (tBu), 26.9 (CH2), 26.8
(CH2). – MS (ESI, 4.1 kV, 8 µl/min, N2, methanol): m/z =
N , methanol): m/z = 145 (4%) [(M-Boc)H] , 189 (27%)
2
i
+
+
[(M- butene)H] , 245 (100%) [MH] . – Analysis for
+
i
+
2
(
01 (19%) [(M-Boc)H] , 245 (30%) [(M- butene)H] , 301
C H N O (244.37): C 63.89, H 11.55, N 11.46; found
1
3 28 2 2
+
100%) [MH] . – Analysis for C H N O (300.49):
1
36
7 2 2
C 63.81, H 11.80, N 11.26.
C 67.95, H 12.08, N 9.32; found C 67.68, H 12.32, N 9.31.
tert-Butyl N-(9-aminononyl)carbamate (9)
tert-Butyl N-{2-[9-(6-chloro-2-methoxyacridinyl)amino]-
Following GP1 from 1,9-diaminononane (15.0 g, ethyl}-carbamate (12)
5 mmol) 9 (4.7 g, 38%) was obtained as an amorphous
9
A mixture of 6,9-dichloro-2-methoxyacridine (1) (4.00 g,
1
white solid. H NMR (400 MHz, CDCl ): δ = 4.52 (br s,
3
1
9
4.4 mmol) and phenol (9 g, 95.6 mmol) was stirred at
0 C for 30 min, then 2 (2.65 g, 16.6 m˜ mol) was added
1
2
2
5
H, NH), 3.07 – 3.05 (m, 2 H, CH (9)), 2.79 – 2.77 (m,
◦
2
H, CH (1)), 1.59 – 1.52 (m, 2 H, CH (2)), 1.42 – 1.40 (m,
2
2
and stirring continued for another 15 min. Chromatographic
work up (silica gel, methanol/ethyl acetate 1:6) yielded 12
H, CH (8)), 1.41 (s, 9 H, tBu), 1.35 – 1.26 (br s, 10 H,
2
13
× CH (3-7)). – C NMR (100 MHz, CDCl ): δ = 155.9
2
3
(1.11 g, 19%) as an amorphous orange-coloured solid. UV-
(
CO), 80.0 (tBu), 41.9 (CH ), 40.7 (CH ), 32.9 (CH ), 30.2
2 2 2
vis (methanol): λmax (log ε) = 295 nm (4.67). – IR (KBr):
ν = 3441s, 2978m, 1678s, 1630s, 1589s, 1367m, 1271s,
(
CH ), 29.6 (CH ), 29.4 (CH ), 29.3 (CH ), 28.6 (tBu),
2 2 2 2
2
6.9 (CH ), 26.8 (CH ). – MS (ESI, 4.1 kV, 8 µl/min,
−1
1
2
2
1
248s, 1171s, 1090m, 1032m cm . – H NMR (400 MHz,
+
N , methanol): m/z = 159 (3%) [(M-Boc)H] , 203 (11%)
3
2
CDCl ): δ = 8.06 (d, JH,H = 9.44 Hz, 1 H, 8-H), 7.81
3
i
+
+
[(M- butene)H] , 259 (100%) [MH] . – Analysis for
4
3
(d, J
= 2.08 Hz, 1 H, 5-H), 7.77 (d, J
= 9.23 Hz,
H,H
H,H
C H N O (258.40): C 65.07, H 11.70, N 10.84; found
4
1
4 30 2 2
1
H, 4-H), 7.43 (d,
J
= 2.49 Hz, 1 H, 1-H), 7.18
dd, JH,H = 9.23 Hz, JH,H = 2.49 Hz, 1 H, 3-H), 7.04
= 2.08 Hz, 1 H, 7-H), 6.13
br s, 1 H, NH), 4.04 – 3.99 (m, 2 H, CH (1’)), 3.93 (s,
H,H
C 64.86, H 11.81, N 10.63.
3
4
(
(
(
dd, 3J
= 9.44 Hz, 4J
H,H
H,H
tert-Butyl N-(10-aminodecyl)carbamate (10)
2
To a solution of 1,10-diaminodecane (15.0 g, 87 mmol) 3 H, OCH3), 3.68 – 3.61 (m, 2 H, CH2(2’)), 1.45 (s, 9 H,
◦
13
in dry dioxane (200 ml) at 0 C a solution of di-tert-butyl- tBu). – C NMR (100 MHz, CDCl3): δ = 157.7 (C=O),
dicarbonate (9.5 g, 44 mmol) in dry dioxane (200 ml) was 155.9 (quart.), 153.3 (quart.), 142.5 (quart.), 138.3 (quart.),
added during 5 h. Then the solvent was removed under re- 138.1 (quart.), 126.3 (CH), 126.0 (CH), 124.0 (CH), 123.4
duced pressure and the residue was extracted with ether (CH), 121.2 (CH), 114.9 (quart.), 111.3 (quart.), 101.4 (CH),
(
3 × 150 ml). The ether was removed and 10 (6.7 g, 57%) 80.1 (tBu), 55.8 (OCH3), 51.2 (CH2(1’)), 41.0 (CH2(2’)),
1
was obtained as white amorphous solid. H NMR (400 MHz, 28.4 (tBu). – MS (ESI, 4.1 kV, 8 µl/min, N2, methanol):
i
+
35
+
CDCl ): δ = 4.50 (br s, 1 H, NH), 3.09 – 3.04 (m, 2 H, 346 (16%) [(M- butene)H] , 402 (100%) [M( Cl)H] ,
3
3
7
+
CH (10)), 2.66 – 2.63 (m, 2 H, CH (1)), 1.41 (s, 9 H, tBu), 404 (27%) [M( Cl)H] . – Analysis for C21H24ClN2O2
2
2
1
.40 – 1.36 (m, 4 H, 2 × CH (2,9)), 1.26 (br s, 12 H, 6 × (401.90): C 62.76, H 6.02, N 10.46; found C 62.35, H 6.17,
2
1
3
CH (3-8)). – C NMR (100 MHz, CDCl ): δ = 155.8 (CO), N 10.22.
7
(
2
8
(
2
3
8.9 (tBu), 42.3 (CH ), 40.6 (CH ), 33.9 (CH ), 30.1
2 2 2
tert-Butyl N-{3-[9-(6-chloro-2-methoxyacridinyl)amino]-
propyl}-carbamate (13)
CH ), 29.6 (CH ), 29.5 (CH ), 29.5 (CH ), 29.3 (CH ),
2 2 2 2 2
8.5 (tBu), 26.9 (CH ), 26.8 (CH ). – MS (ESI, 4.1 kV,
2 2
+
µl/min, N , methanol): m/z = 173 (5%) [(M-Boc)H] , 217
Following the procedure given for 12 from 1 (3.0 g,
2
i
+
+
16%) [(M- butene)H] , 273 (100%) [MH] . – Analysis for 10.8 mmol), phenol (9.0 g, 95.6 mmol) and 3 (2.3 g,
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93
1
3.2 mmol) 13 (3.59 g, 80%) was obtained as a yel- 28.7 mmol) 16 (5.63 g, 51%) was obtained as a yellow amor-
low amorphous solid. UV-vis (methanol): λmax (log ε) = phous solid. MS (ESI, 4.1 kV, 8 µl/min, N , methanol):
2
i
+
35
+
2
3
4
82 nm (4.72). – MS (ESI, 4.1 kV, 8 µl/min, N , methanol): 402 (7%) [(M- butene)H] , 458 (100%) [M( Cl)H] ,
2
i
+
35
+
37
+
60 (13%) [(M- butene)H] , 416 (100%) [M( Cl)H] , 460 (37%) [M( Cl)H] . – Analysis for C H ClN O
2 2
2
5
32
3
7
+
18 (30%) [M( Cl)H] . – Analysis for C H ClN O
2 2
(458.01): C 65.56, H 7.04, N 9.18; found C 65.37, H 7.26,
22
26
(415.92): C 63.53, H 6.30, N 10.10; found C 63.26, H 6.55, N 9.11.
N 10.3.
tert-Butyl N-{7-[9-(6-chloro-2-methoxyacridinyl)amino]-
heptyl}-carbamate (17)
tert-Butyl N-{4-[9-(6-chloro-2-methoxyacridinyl)amino]-
butyl}-carbamate (14)
Following the procedure given for 12 from 1 (3.16 g,
Following the procedure given for 12 from 1 (3.0 g, 11.4 mmol), phenol (9.0 g, 95.6 mmol) and 7 (2.87 g,
0.8 mmol), phenol (9.0 g, 95.6 mmol) and 4 (2.43 g, 12.5 mmol) 17 (2.98 g, 56%) was obtained as a yellow amor-
1
1
2.9 mmol) 14 (2.53 g, 55%) was obtained as a yellow amor- phous solid. MS (ESI, 4.1 kV, 8 µl/min, N , methanol):
2
+
i
+
phous solid. MS (ESI, 4.1 kV, 8 µl/min, N , methanol): 372 (6%) [(M-Boc)H] , 416 (12%) [(M- butene)H] , 472
2
i
+
35
+
35
+
37
+
3
4
74 (9%) [(M- butene)H] , 430 (100%) [M( Cl)H] , (100%) [M( Cl)H] , 474 (34%) [M( Cl)H] . – Analysis
37
+
32 (32%) [M( Cl)H] . – Analysis for C H ClN O
for C H ClN O (472.03): C 66.16, H 7.26, N 8.90; found
23
28
2
2
26 34 2 2
(429.95): C 64.25, H 6.56, N 9.77; found C 64.01, H 6.68, C 66.01, H 7.39, N 8.78.
N 9.58.
tert-Butyl N-{8-[9-(6-chloro-2-methoxyacridinyl)amino]-
octyl}-carbamate (18)
tert-Butyl N-{5-[9-(6-chloro-2-methoxyacridinyl)amino]-
pentyl}-carbamate (15)
Following the procedure given for 12 from 1 (4.7 g,
Following the procedure given for 12 from 1 (3.2 g, 16.9 mmol), phenol (14.0 g, 0.15 mol) and 8 (4.6 g,
1.5 mmol), phenol (9.0 g, 95.6 mmol) and 5 (2.89 g, 18.6 mmol) 18 (4.00 g, 49%) was obtained as a yellow amor-
1
1
4.3 mmol) 15 (4.34 g, 85%) was obtained as a yel- phous solid. MS (ESI, 4.1 kV, 8 µl/min, N , methanol):
2
+
i
+
low amorphous solid. UV-vis (methanol): λmax (log ε) = 386 (3%) [(M-Boc)H] , 430 (7%) [(M- butene)H] , 486
3
5
+
37
+
2
1
1
83 nm (4.75). – IR (KBr): ν = 3432s, 2932s, 1689s, (100%) [M( Cl)H] , 488 (38%) [M( Cl)H] . – Analysis
631s, 1562s, 1520s, 1466s, 1436m, 1391m, 1365m, 1238s, for C H ClN O (486.06): C 66.72, H 7.47, N 8.65; found
2
7
36
2 2
1
170s, 1033m. – H NMR (400 MHz, CDCl ): δ = 8.03 C 66.51, H 7.63, N 8.42.
3
4
3
(
d, J
= 2.08 Hz, 1 H, 5-H), 8.00 (d, J
= 9.34 Hz,
H,H
H,H
3
1
H, 8-H), 7.95 (d, J
= 9.44 Hz, 1 H, 4-H), 7.36 (dd,
H,H
tert-Butyl N-{9-[9-(6-chloro-2-methoxyacridinyl)amino]-
nonyl}-carbamate (19)
3
JH,H = 9.44 Hz, 4JH,H = 2.59 Hz, 1 H, 3-H), 7.27 (dd,
JH,H = 9.34 Hz, 4JH,H = 2.08 Hz, 1 H, 7-H), 7.20 (d,
3
4J
3
2
(
(
= 2.59 Hz, 1 H, 1-H), 4.53 (br s, 1 H, NH), 3.94 (s,
Following the procedure given for 12 from 1 (2.5 g,
H, OCH ), 3.71 – 3.65 (m, 2 H, CH (1’)), 3.14 – 3.07 (m, 9.0 mmol), phenol (9.0 g, 95.6 mmol) and 9 (2.97 g,
H,H
3
2
H, CH (5’)), 1.81 – 1.73 (m, 2 H, CH (2’)), 1.53 – 1.43 11.5 mmol) 19 (2.16 g, 48%) was obtained as an amorphous
2
2
13
m, 4 H, 2 × CH (3’,4’)), 1.41 (s, 9 H, tBu). – C NMR yellow solid. MS (ESI, 4.1 kV, 8 µl/min, N2, methanol):
2
+
i
+
100 MHz, CDCl ): δ = 155.91 (C=O), 155.88 (quart.), 400 (7%) [(M-Boc)H] , 444 (17%) [(M- butene)H] , 500
3
3
5
+
37
+
1
1
49.8 (quart.), 147.6 (quart.), 145.9 (quart.), 134.9 (quart.), (100%) [M( Cl)H] , 502 (33%) [M( Cl)H] . – Analysis
30.7 (CH), 127.5 (CH), 124.5 (CH), 124.4 (CH), 124.1 for C28H38ClN2O2 (500.09): C 67.25, H 7.66, N 8.40; found
(
CH), 117.7 (quart.), 115.5 (quart.), 99.4 (CH), 79.2 (tBu), C 67.02, H 7.86, N 8.25.
5
3
8
5.6 (OCH ), 50.5 (CH (1’)), 40.2 (CH (5’)), 31.3 (CH ),
3 2 2 2
0.0 (CH ), 28.5 (tBu), 24.2 (CH ). – MS (ESI, 4.1 kV, tert-Butyl N-{3-[10-(6-chloro-2-methoxyacridinyl)amino]-
2
2
i
+
µl/min, N , methanol): 388 (10%) [(M- butene)H] , 444 decyl}-carbamate (20)
2
3
5
+
37
+
(100%) [M( Cl)H] , 446 (30%) [M( Cl)H] . – Analysis
Following the procedure given for 12 from 1 (2.7 g,
.7 mmol), phenol (9.0 g, 95.6 mmol) and 10 (2.7 g,
.9 mmol) 20 (3.04 g, 61%) was obtained as an amorphous
for C H ClN O (443.98): C 64.93, H 6.81, N 9.46; found
C 64.85, H 7.02, N 9.29.
2
4
30
2 2
9
9
yellow solid. MS (ESI, 4.1 kV, 8 µl/min, N , methanol):
2
tert-Butyl N-{6-[9-(6-chloro-2-methoxyacridinyl)amino]-
hexyl}-carbamate (16)
+
i
+
4
14 (7%) [(M-Boc)H] , 458 (12%) [(M- butene)H] , 514
3
5
+
37
+
(
100%) [M( Cl)H] , 516 (33%) [M( Cl)H] . – Analysis
Following the procedure given for 12 from 1 (6.65 g, for C H ClN O (514.11): C 67.75, H 7.84, N 8.17; found
2
9
40
2 2
2
3.9 mmol), phenol (18.0 g, 0.2 mol) and 6 (6.2 g, C 67.51, H 7.99, N 8.12.
Unauthenticated
Download Date | 10/8/18 1:47 PM

9
4
Ren e´ Csuk et al. · Synthesis of Dimeric Acridine Derived Nucleic Acid Intercalators
1
tert-Butyl N-{12-[9-(6-chloro-2-methoxyacridinyl)amino]-
dodecyl}-carbamate (21)
N -[9-(6-Chloro-2-methoxyacridinyl)]-1,3-propane-
diamine bis(hydrochloride) (23)
Following the procedure given for 12 from 1 (5.56 g,
A solution of 13 (3.39 g, 8.1 mmol) in methanol (100 ml)
2
2
0.0 mmol), phenol (18.0 g, 0.2 mol) and 11 (7.2 g, containing aq. hydrochloric acid (10%,10 ml) was stirred
4.0 mmol) 21 (2.80 g, 26%) was obtained as a yel- at room temperature for 16 h. After evaporation of the sol-
low amorphous solid. UV-vis (methanol): λmax (log ε) = vents 23 (3.07 g, 97%) was obtained as a yellow amorphous
2
1
1
83 nm (4.58). IR (KBr): ν = 3367m, 2926s, 2853s, 2711m, solid. MS (ESI, 4.1 kV, 8 µl/min, N2, methanol): 316 (100%)
35 37
+
+
688s, 1631s, 1562s, 1520s, 1467s, 1436s, 1392m, 1364s, [M( Cl)H] , 318 (30%) [M( Cl)H] .
−
1
1
273s, 1244s, 1171s, 1074m, 1032m cm . – H NMR
4
1
(
400 MHz, CDCl ): δ = 7.98 (d, JH,H = 2.08 Hz, 1 H, N -[9-(6-Chloro-2-methoxyacridinyl)]-1,4-butane-diamine
3
3
3
5
9
-H), 7.98 (d, J
= 9.13 Hz, 1 H, 4-H), 7.87 (d, J
=
bis(hydrochloride) (24)
H,H
H,H
.34 Hz, 1 H, 8-H), 7.29 – 7.24 (m, 2 H, H-C(3,1)), 7.22
A solution of 14 (2.43 g, 5.7 mmol) in methanol (100 ml)
containing aq. hydrochloric acid (10%, 10 ml) was stirred
at room temperature for 16 h. After evaporation of the sol-
vents 24 (2.18 g, 96%) was obtained as a yellow amorphous
solid. MS (ESI, 4.1 kV, 8 µl/min, N , methanol): 330 (100%)
M( Cl)H] , 332 (32%) [M( Cl)H] .
dd, JH,H = 9.34 Hz, 4JH,H = 2.08 Hz, 1 H, 7-H), 4.49
3
(
(
2
1
br s, 1 H, NH), 3.94 (s, 3 H, OCH ), 3.78 – 3.73 (m,
H, CH (1’)), 3.10 – 3.04 (m, 2 H, CH (12’)), 1.85 –
2 2
3
.76 (m, 2 H, CH (2’)), 1.47 – 1.39 (m, 13 H, tBu, 2 ×
2
2
CH (3’,11’)), 1.35 – 1.19 (m, 14 H, 5 × CH (4’-10’)). –
2
2
35
+
37
+
[
1
3
C NMR (100 MHz, CDCl ): δ = 155.8 (C=O), 155.7
3
(
(
5
3
2
(
4
(
quart.), 150.9 (quart.), 140.1 (quart.), 136.0 (quart.), 124.8
1
N -[9-(6-Chloro-2-methoxyacridinyl)]-1,5-pentane-diamine
CH), 124.7 (CH), 124.1 (CH), 101.0 (CH), 79.0 (tBu),
bis(hydrochloride) (25)
5.7 (OCH ), 50.2 (CH (1’)), 40.7 (CH (12’)), 31.5 (CH ),
3 2 2 2
0.1 (CH ), 29.57 (2 x CH ), 29.54 (CH ), 29.52 (CH ),
2
2
2
2
A solution of 15 (4.19 g, 9.4 mmol) in methanol (100 ml)
containing aq. hydrochloric acid (10%, 10 ml) was stirred
at room temperature for 16 h. After evaporation of the sol-
vents 25 (3.65 g, 93%) was obtained as a yellow amor-
phous solid. UV-vis (methanol): λmax (log ε) = 294 nm
(4.36). – IR (KBr): ν = 3417s, 2965s, 2004w, 1627s,
9.37 (CH ), 29.32 (CH ), 28.5 (tBu), 27.0 (CH ), 26.9
2
2
2
CH ). – MS (ESI, 4.1 kV, 8 µl/min, N , methanol):
2
2
+
i
+
42 (7%) [(M-Boc)H] , 487 (15%) [(M- butene)H] , 542
3
5
+
37
+
100%) [M( Cl)H] , 544 (34%) [M( Cl)H] . – Analysis
for C H ClN O (542.17): C 68.68, H 8.18, N 7.75; found
3
1
44
2 2
C 68.55, H 8.29, N 7.53.
1
1
590s, 1567s, 1524m, 1497s, 1483s, 1420m, 1396m, 1358m,
318m, 1292m, 1267m, 1245s, 1170m, 1120m, 1094m,
1
−1
1
N -[9-(6-Chloro-2-methoxyacridinyl)]-1,2-ethane-diamine
1029m cm . – H NMR (400 MHz, CD OD): δ = 7.67
3
3
3
bis(hydrochloride) (22)
(d, J
= 9.34 Hz, 1 H, 8-H), 7.21 (dd, J
= 9.34 Hz,
H,H
H,H
4
JH,H = 2.49 Hz, 1 H, 3-H), 7.10 – 7.02 (m, 3 H, H-C(1,4,7)),
A solution of 12 (1.06 g, 2.6 mmol) in methanol (100 ml)
containing aq. hydrochloric acid (10%, 10 ml) and trifluo-
roacetic acid (2 ml) was stirred for 16 h at room temperature.
After evaporation of the solvents 22 (0.93 g, 94%) was ob-
tained as a yellow amorphous solid. UV-vis (methanol): λmax
4
6
3
1
1
.90 (d, J
= 2.08 Hz, 1 H, 5-H), 3.69 (s, 3 H, OCH3),
H,H
.66 – 3.60 (m, 2 H, CH (1’)), 2.89 – 2.83 (m, 2 H, CH (5’)),
.73 – 1.65 (m, 2 H, CH (2’)), 1.62 – 1.54 (m, 2 H, CH (4’)),
.36 – 1.28 (m, 2 H, CH (3’)). – C NMR (100 MHz, D O):
2
2
2
2
13
2
2
δ = 155.5 (quart.), 154.3 (quart.), 140.4 (quart.), 138.2
quart.), 133.1 (quart.), 126.8 (CH), 124.3 (CH), 119.7 (CH),
16.7 (CH), 112.3 (quart.), 108.6 (quart.), 56.2 (OCH ), 48.5
(
log ε) = 292 nm (4.66). – IR (KBr): ν = 3426s, 2967s,
(
1
1
629s, 1590s, 1530m, 1500m, 1470m, 1446m, 1397w,
1
3
−1
373w, 1275m, 1249m, 1174m, 1094m, 1028w cm . –
(
(
(
CH (1’)), 39.5 (CH (5’)), 29.1 (CH ), 26.7 (CH ), 23.3
2 2 2 2
1
3
H NMR (400 MHz, CD OD): δ = 8.48 (d, J
= 9.34 Hz,
3
H,H
CH ). – MS (ESI, 4.1 kV, 8 µl/min, N , methanol): 344
2
2
1
H, 8-H), 7.96 (d, 4JH,H = 2.49 Hz, 1 H, 1-H), 7.86 (d,
35
+
37
+
100%) [M( Cl)H] , 346 (30%) [M( Cl)H] .
4
3
JH,H = 2.08 Hz, 1 H, 5-H), 7.79 (d, J
= 9.34 Hz, 1 H,
H,H
3
4
4
3
7
3
-H), 7.66 (dd, JH,H = 9.34 Hz, JH,H = 2.49 Hz, 1 H,
1
N -[9-(6-Chloro-2-methoxyacridinyl)]-1,6-hexane-diamine
-H), 7.53 (dd, 3JH,H = 9.34 Hz, 4JH,H = 2.08 Hz, 1 H,
bis(hydrochloride) (26)
-H), 4.54 – 4.50 (m, 2 H, CH (1’)), 4.05 (s, 3 H, OCH ),
2
3
13
.64 – 3.59 (m, 2 H, CH (2’)). – C NMR (100 MHz,
A solution of 16 (6.50 g, 14.2 mmol) in methanol (100 ml)
2
CD OD): δ = 157.6 (quart.), 157.5 (quart.), 141.9 (quart.), containing aq. hydrochloric acid (10%, 10 ml) and triflu-
3
1
(
1
40.7 (quart.), 135.4 (quart.), 128.8 (CH), 128.3 (CH), 125.6 oroacetic acid (2 ml) was stirred at room temperature for
CH), 121.3 (CH), 118.4 (CH), 115.2 (quart.), 110.8 (quart.), 16 h. After evaporation of the solvents 26 (5.98 g, 98%) was
03.6 (CH), 57.2 (OCH ), 46.9 (CH (1’)), 39.7(CH (2’)). – obtained as an amorphous yellow solid. MS (ESI, 4.1 kV,
3
2
2
˜
35
+
MS (ESI, 4.1kV, 8 µl/min, N , methanol): 302 (100%) 8 µl/min, N , methanol): 358 (100%) [M( Cl)H] , 360
2
2
3
5
+
37
+
37
+
[M( Cl)H] , 304 (30%) [M( Cl)H] .
(32%) [M( Cl)H] .
Unauthenticated
Download Date | 10/8/18 1:47 PM

Ren e´ Csuk et al. · Synthesis of Dimeric Acridine Derived Nucleic Acid Intercalators
95
1
1
N -[9-(6-Chloro-2-methoxyacridinyl)]-1,7-heptane-diamine
1273s, 1244s, 1171s, 1074m, 1032m. – H NMR (400 MHz,
3
bis(hydrochloride) (27)
CD OD): δ = 8.46 (d, JH,H = 9.28 Hz, 1 H, 8-H), 7.83
3
4
4
(
1
(
(
d, J
= 2.44 Hz, 1 H, 1-H), 7.80 (d, J
= 1.95 Hz,
H,H
H,H
A solution of 17 (3.11 g, 6.6 mmol) in methanol (100 ml)
containing aq. hydrochloric acid (10%, 10 ml) and triflu-
oroacetic acid (3 ml) was stirred at room temperature for
6 h. After evaporation of the solvents 27 (2.77 g, 95%)
was obtained as a yellow amorphous solid. MS (ESI, 4.1 kV,
3
H, 5-H), 7.76 (d,
J
= 9.28 Hz, 1 H, 4-H), 7.67
= 2.44 Hz, 1 H, 3-H), 7.50
= 1.95 Hz, 1 H, 7-H), 4.16 –
H,H
dd, JH,H = 9.28 Hz, 4J
3
H,H
3
4
dd, J
= 9.28 Hz, J
H,H
H,H
1
4
2
1
.11 (m, 2 H, CH (1’)), 4.00 (s, 3 H, OCH ), 2.93 –
.87 (m, 2 H, CH (12’)), 2.02 – 1.93 (m, 2 H, CH (2’)),
2
3
2
2
35
+
8
(
µl/min, N , methanol): 372 (100%) [M( Cl)H] , 374
2
31%) [M( Cl)H] .
.68 – 1.59 (m, 2 H, CH (11’)), 1.51 – 1.27 (m, 16H, 8 ×
2
3
7
+
13
CH (3’ – 10’)). – C NMR (125 MHz, CD OD): δ =
2
3
158.2 (quart.), 158.0 (quart.), 155.7 (quart.), 141.8 (quart.),
141.5 (quart.), 135.8 (quart.), 129.2 (CH), 128.7 (CH),
125.0 (CH), 121.4 (CH), 118.4 (CH), 115.5 (quart.), 111.3
1
N -[9-(6-Chloro-2-methoxyacridinyl)]-1,8-octane-diamine
bis(hydrochloride) (28)
(
quart.), 104.1, 79.0 (tBu), 55.7 (OCH ), 50.2 (CH (1’)),
3 2
A solution of 18 (3.90 g, 8.0 mmol) in methanol (100 ml)
containing aq. hydrochloric acid (10%, 10 ml) was stirred
at room temperature for 16 h. After evaporation of the sol-
vents 28 (3.52 g, 96%) was obtained as a yellow amor-
4
2
2
8
0.7 (CH (12’)), 31.5 (CH ), 30.1 (CH ), 29.57 (CH ),
2 2 2 2
9.54 (CH ), 29.52 (CH ), 29.37 (CH ), 29.32 (CH ),
2
2
2
2
8.5 (tBu), 27.0 (CH ), 26.9 (CH ). – MS (ESI, 4.1 kV,
2
2
+
µl/min, N , methanol): 442 (7%) [(M-Boc)H] , 487 (15%)
2
phous solid. MS (ESI, 4.1 kV, 8 µl/min, N , methanol):
2
i
+
35
+
3
5
2+
37
2+
[(M- butene)H] , 542 (100%) [M( Cl)H] , 544 (34%)
[M( Cl)H] .
1
(
94 (30%) [M( Cl)H ] , 195 (11%) [M( Cl)H ] , 386
2
2
3
7
+
3
5
+
37
+
100%) [M( Cl)H] , 388 (34%) [M( Cl)H] .
1
N -[9-(6-Chloro-2-methoxyacridinyl)]-1,9-nonane-diamine
2-((2-{[2-(9-{6-Chloro-2-methoxyacridinyl}amino)ethyl]-
amino}-2-oxoethyl){2-[(2-{[2-(9-{6-chloro-2-methoxy-
acridinyl}amino) ethyl]amino}-2-oxoethyl)(carboxymeth-
yl)amino]ethyl}amino) acetic acid (32)
bis(hydrochloride) (29)
A solution of 19 (1.91 g, 3.8 mmol) in methanol (100 ml)
containing aq. hydrochloric acid (10%, 10 ml) was stirred
at room temperature for 16 h. After evaporation of the
solvents 29 (1.71 g, 95%) was obtained as an amorphous
yellow solid. MS (ESI, 4.1 kV, 8 µl/min, N , methanol):
01 (38%) [M( Cl)H ] , 202 (13%) [M( Cl)H ] , 400
A solution of 22 (500 mg, 1.34 mmol) in dry DMF
(
50 ml) containing triethylamine (3 ml) is stirred for 30 min
at room temperature and EDTA dianhydride (154 mg,
.60 mmol) is added and stirring is continued for an-
2
3
5
2+
37
2+
2
(
2
2
0
3
5
+
37
+
100%) [M( Cl)H] , 402 (32%) [M( Cl)H] .
other 15 h. After addition of acetone (10 ml) the precip-
itate is filtered off, washed with methanol (50 ml) and
dried in vacuum to afford 32 (140 mg, 27%) as an or-
ange amorphous solid. UV-vis (methanol): λmax (log ε) =
1
N -[9-(6-Chloro-2-methoxyacridinyl)]-1,10-decane-
diamine bis(hydrochloride) (30)
2
1
1
96 nm (4.86). – IR (KBr): ν = 3423s, 2936m, 1630s,
A solution of 20 (2.89 g, 5.6 mmol) in methanol (100 ml)
containing aq. hydrochloric acid (10%, 10 ml) and tri-
fluoroacetic acid (2 ml) was stirred at room temperature
for 16 h. After evaporation of the solvents 30 (2.68 g,
8%) was obtained as a yellow amorphous solid. MS (ESI,
.1 kV, 8 µl/min, N , methanol): 208 (73%) [M( Cl)H ]
09 (23%) [M( Cl)H ] , 414 (100%) [M( Cl)H] , 416
588s, 1500m, 1475m, 1396m, 1249m, 1172m, 1090m,
−
1
1
030w cm . – H NMR (400 MHz, CD OD): δ = 8.44
3
3
(d, J
= 9.34 Hz, 2 H, 2 × 8-H), 7.69 (s, 2 H, 2 × 1-H),
H,H
.62 (d,
3
7
J
H,H
= 9.34 Hz, 2 H, 2 × 4-H), 7.61 (s, 2 H,
3
9
4
2
35
2+
2 × 5-H), 7.54 (d,
J
H,H
= 9.34 Hz, 2 H, 2 × 3-H), 7.38
,
2
2
3
3
7
2+
35
+
(d,
J
H,H
= 9.34 Hz, 2 H, 2 × 7-H), 4.27 – 4.23 (m, 4 H,
2
3
7
+
2×CH2(1’)), 3.98 (s, 6 H, 2×OCH3), 3.78 – 3.74 (m, 4 H,
(
32%) [M( Cl)H] .
2
× CH (2’)), 3.67 (s, 4 H, 2 × CH (2”)), 3.56 (s, 4 H, 2 ×
2
2
13
1
CH2(3”)), 3.09 (s, 4 H, 2×CH2(1”)). – C NMR (100 MHz,
N -[9-(6-Chloro-2-methoxyacridinyl)]-1,12-dodecane-
CD OD): δ = 173.3 (C=O), 173.1 (C=O), 158.1 (quart.),
3
diamine bis(hydrochloride) (31)
1
41.8 (quart.), 128.6 (CH), 125.4 (CH), 121.3 (CH), 118.4
A solution of 21 (2.75 g, 5.1 mmol) in methanol (100 ml) (CH), 111.3 (quart.), 58.3 (CH (3”)), 57.4 (CH (2”)), 57.0
2
2
containing aq. hydrochloric acid (10%, 10 ml) was stirred (OCH ), 53.7 (CH (1”)), 48.0 (CH (1’)), 40.7 (CH (2’)). –
3
2
2
2
at room temperature for 16 h. After evaporation of the MS (ESI, 4.1 kV, 8 µl/min, N , methanol): 431 (42%)
2
2
+
35
35
+
solvents 31 (2.53 g, 97%) was obtained as an amorphous [MH ] , 859 (79%) [M( Cl)( Cl)H] , 861 (100%)
yellow solid. UV-vis (methanol): λmax (log ε) = 283 nm [M( Cl)( Cl)H] , 863 (23%) [M( Cl)( Cl)H] . – Anal-
4.58). – IR (KBr): ν = 3367m, 2926s, 2853s, 2711m, ysis for C H Cl N O (859.75): C 58.68, H 5.16, N 8.25;
2
3
7
35
+
37
37
+
(
1
42 44 2 8 8
688s, 1631s, 1562s, 1520s, 1467s, 1436s, 1392m, 1364s, found C 58.42, H 5.23, N 8.11.
Unauthenticated
Download Date | 10/8/18 1:47 PM

9
6
Ren e´ Csuk et al. · Synthesis of Dimeric Acridine Derived Nucleic Acid Intercalators
1
3
2
-((2-{[3-(9-{6-Chloro-2-methoxyacridinyl}amino)propyl]-
C NMR (100 MHz, CD OD): δ = 172.4 (C=O), 170.1
3
amino}-2-oxoethyl){2-[(2-{[3-(9-{6-chloro-2-methoxy-
acridinyl}amino) propyl]amino}-2-oxoethyl)(carboxy-
methyl)amino]ethyl}amino) acetic acid (33)
(C=O), 157.8 (quart.), 141.8 (quart.), 128.5 (CH), 125.0
(CH), 121.4 (CH), 118.4 (CH), 58.2 (CH (3”)), 56.84
(OCH ), 56.80 (CH (2”)), 53.2 (CH (1”)), 50.3 (CH (1’)),
4
2
3
2
2
2
0.0 (CH (5’)), 30.4 (CH ), 29.9 (CH ), 25.1 (CH ). – MS
2 2 2 2
Following the procedure for 32 from 23 (0.8 g,
.06 mmol) and EDTA dianhydride (237 mg, 0.93 mmol)
2+
(
4
ESI, 4.1 kV, 8 µl/min, N , methanol): 473 (32%) [MH ]
,
2
2
2
2+
2+
92 (100%) [MHK] , 500 (34%) [MNaK] , 943 (42%)
in dry DMF (35 ml) and triethylamine (3 ml) 33 (425 mg,
35
35
+
37
35
+
[
(
[
M( Cl)( Cl)H] , 945 (62%) [M( Cl)( Cl)H] , 947
16%) [M( Cl)( Cl)H] , 965 (12%) [MNa] , 981 (30%)
MK] . – Analysis for C H Cl N O (943.94): C 61.08,
5
4
4
2%) was obtained as an amorphous orange solid. MS (ESI,
37
37
+
+
2
+
.1 kV, 8 µl/min, N , methanol): 445 (92%) [MH ]
,
+
2
2
4
8
56
2 8 8
2+
2+
64 (71%) [MHK] , 471 (38%) [MNaK] , 887 (85%)
H 5.98, N 11.87; found C 59.95, H 6.14, N 11.65.
3
5
35
+
37
35
+
[
(
[
M( Cl)( Cl)H] , 889 (100%) [M( Cl)( Cl)H] , 891
3
7
37
+
+
22%) [M( Cl)( Cl)H] , 909 (10%) [MNa] , 925 (20%) 2-((2-{[6-(9-{6-Chloro-2-methoxyacridinyl}amino)hexyl]-
+
MK] . – Analysis for C44H Cl N O (887.83): C 59.53, amino}-2-oxoethyl){2-[(2-{[6-(9-{6-chloro-2-methoxy-
48
2 8 8
H 5.45, N 7.99; found C 59.29, H 5.62, N 7.84.
acridinyl}amino) hexyl]amino}-2-oxoethyl)(carboxymeth-
yl)amino]ethyl}amino) acetic acid (36)
2
-((2-{[4-(9-{6-Chloro-2-methoxyacridinyl}amino)butyl]-
Following the procedure for 32 from 26 (1.5 g, 3.5 mmol)
amino }-2-oxoethyl){2-[(2-{[4-(9-{6-chloro-2-methoxy-
acridinyl}amino) butyl]amino}-2-oxoethyl)(carboxymeth-
yl)amino]ethyl}amino) acetic acid (34)
and EDTA dianhydride (405 mg, 1.58 mmol) in DMF
(30 ml) and triethylamine (5 ml) 36 (980 mg, 64%) was
obtained as an orange coloured amorphous solid. MS (ESI,
4
5
2
+
.1 kV, 8 µl/min, N , methanol): 487 (25%) [MH ]
06 (56%) [MHK] , 514 (31%) [MNaK] , 971 (100%)
,
2
2
Following the procedure for 32 from 24 (1.0 g, 2.5 mmol)
2
+
2+
and EDTA dianhydride (287 mg, 1.12 mmol) in dry DMF
3
5
35
+
37
35
+
[M( Cl)( Cl)H] , 973 (90%) [M( Cl)( Cl)H] , 975
(60 ml) and triethylamine (5 ml) 34 (412 mg, 40%)
3
7
37
+
+
(20%) [M( Cl)( Cl)H] , 993 (10%) [MNa] , 1009 (30%)
was obtained as an orange amorphous solid. MS (ESI,
+
2
+
[MK] . – Analysis for C50H60Cl2N8O8 (971.99): C 61.79,
H 6.22, N 11.53; found C 61.54, H 6.39, N 11.42.
4
4
.1 kV, 8 µl/min, N , methanol): 459 (22%) [MH ]
,
2
2
2+
2+
78 (78%) [MHK] , 486 (40%) [MNaK] , 915 (82%)
3
5
35
+
37
35
+
[
(
[
M( Cl)( Cl)H] , 917 (100%) [M( Cl)( Cl)H] , 919
20%) [M( Cl)( Cl)H] , 937 (10%) [MNa] , 953 (30%)
MK] . – Analysis for C46H Cl N O (915.88): C 60.33,
2
-((2-{[7-(9-{6-Chloro-2-methoxyacridinyl}amino)heptyl]-
3
7
37
+
+
amino}-2-oxoethyl){2-[(2-{[7-(9-{6-chloro-2-methoxy-
acridinyl}amino) heptyl]amino}-2-oxoethyl)(carboxy-
methyl)amino]ethyl}amino) acetic acid (37)
+
52
2 8 8
H 5.72, N 12.23; found C 60.14, H 5.84, N 12.00.
Following the procedure for 32 from 27 (0.5 g,
.13 mmol) and EDTA dianhydride (144 mg, 0.68 mmol)
in DMF (10 ml) and triethylamine (3 ml) 37 (290 mg,
2%) was obtained as an amorphous orange solid. MS (ESI,
2
-((2-{[5-(9-{6-Chloro-2-methoxyacridinyl}amino)pentyl]-
1
amino}-2-oxoethyl){2-[(2-{[5-(9-{6-chloro-2-methoxy-
acridinyl}amino) pentyl]amino}-2-oxoethyl)(carboxy-
methyl)amino]ethyl}amino) acetic acid (35)
5
4
2
+
.1 kV, 8 µl/min, N , methanol): 501 (21%) [MH ]
,
2
2
2
+
2+
Following the procedure for 32 from 25 (0.5 g, 1.2 mmol) 520 (87%) [MHK] , 528 (22%) [MNaK] , 1000 (100%)
and EDTA dianhydride (139 mg, 0.54 mmol) in dry DMF [M( Cl)( Cl)H] , 1002 (83%) [M( Cl)( Cl)H] , 1004
50 ml) and triethylamine (5 ml) 35 (280 mg, 55%) was ob- (22%) [M( Cl)( Cl)H] , 1022 (9%) [MNa] , 1038 (28%)
tained as an amorphous orange solid. UV-vis (methanol): [MK] . – Analysis for C52H64Cl2N8O8 (1000.64): C 62.46,
3
5
35
+
37
35
+
3
7
37
+
+
(
+
λmax (log ε) = 298 nm (4.94). – IR (KBr): ν = 3387s, H 6.45, N 11.21; found C 62.39, H 6.61, N 11.02.
2
1
934s, 1724w, 1631s, 1590s, 1501s, 1470m, 1396m, 1358m,
273m, 1245s, 1170m, 1092m, 1029w cm . – H NMR
−1
1
2-((2-{[8-(9-{6-Chloro-2-methoxyacridinyl}amino)octyl]-
amino}-2-oxoethyl){2-[(2-{[8-(9-{6-chloro-2-methoxy-
acridinyl}amino) octyl]amino}-2-oxoethyl)(carboxymethyl)
amino]ethyl}amino) acetic acid (38)
3
(
400 MHz, CD OD): δ = 8.40 (d, J
= 9.34 Hz, 2 H,
3
H,H
4
2
× 8-H), 7.78 (d, J
= 2.59 Hz, 2 H, 2 × 1-H), 7.73 (d,
H,H
4
3
J
= 2.08 Hz, 2 H, 2 × 5-H), 7.70 (d, J
= 9.34 Hz,
= 2.59 Hz,
H,H
H,H
3
4
2
2
2
6
2
2
H, 2×4-H), 7.60 (dd, J
= 9.34 Hz, J
Following the procedure for 32 from 28 (0.7 g,
H,H
H,H
3
4
H, 2×3-H), 7.45 (dd, J
= 9.34 Hz, J
= 2.08 Hz, 1.53 mmol) and EDTA dianhydride (176 mg, 0.69 mmol) in
H,H
H,H
H, 2 × 7-H), 4.12 – 4.07 (m, 4 H, 2 × CH (1’)), 3.98 (s, DMF (35 ml) and triethylamine (3 ml) 38 (320 mg, 45%) was
2
H, 2 × OCH ), 3.93 (s, 4 H, 2 × CH (2”)), 3.86 (s, 4 H, obtained as an amorphous orange coloured solid. MS (ESI,
3
2
2
+
×CH (3”)), 3.34 (s, 4 H, 2×CH (1”)), 3.30 – 3.26 (m, 4 H, 4.1 kV, 8 µl/min, N , methanol): 515 (32%) [MH ] , 534
2
2
2
2
2
+
35
35
+
×CH (5’)), 2.05 – 1.97 (m, 4 H, 2 ×CH (2’)), 1.67 – 1.63 (100%) [MHK] , 1028 (25%) [M( Cl)( Cl)H] , 1030
2
2
3
7
35
+
+
(
m, 4 H, 2 × CH (4’)), 1.56 – 1.49 (m, 4 H, 2 × CH (3’)). – (9%) [M( Cl)( Cl)H] , 1050 (7%) [MNa] , 1066 (42%)
2
2
Unauthenticated
Download Date | 10/8/18 1:47 PM

Ren e´ Csuk et al. · Synthesis of Dimeric Acridine Derived Nucleic Acid Intercalators
97
+
[
MK] . – Analysis for C H Cl N O (1028.09): C 63.09, in DMF (50 ml) and triethylamine (5 ml) 40 (1.04 g,
54
68
2 8 8
H 6.67, N 10.90; found C 62.94, H 6.81, N 10.77.
46%) was obtained as an amorphous orange coloured solid.
UV-vis (methanol, 0.1% TFA): λmax (log ε) = 295 nm
2
-((2-{[9-(9-{6-Chloro-2-methoxyacridinyl}amino)nonyl]-
(4.92). – IR (KBr): ν = 3416s, 2926s, 2853s, 1742m, 1672m,
amino}-2-oxoethyl){2-[(2-{[9-(9-{6-chloro-2-methoxy-
acridinyl}amino) nonyl]amino}-2-oxoethyl)(carboxy-
methyl)amino]ethyl}amino) acetic acid (39)
1
1
628s, 1589s, 1563s, 1500m, 1467m, 1384m, 1361m, 1248s,
171m, 1091m, 1031m cm . – H NMR (400 MHz,
−1
1
3
CD OD): δ = 8.41 (d, JH,H = 9.23 Hz, 2 H, 2 × 8-H),
3
4
4
7
2
2
2
.79 (d, J
= 2.59 Hz, 2 H, 2 × 1-H), 7.77 (d, J
=
Following the procedure for 32 from 29 (1.0 g, 2.1 mmol)
H,H
H,H
3
.08 Hz, 2 H, 2 × 5-H), 7.74 (d,
J
= 9.23 Hz, 2 H,
and EDTA dianhydride (244 mg, 0.95 mmol) in DMF
H,H
3
4
× 4-H), 7.60 (dd,
J
= 9.23 Hz,
= 9.23 Hz, J
J
H,H
= 2.59 Hz,
= 2.08 Hz,
(50 ml) and triethylamine (5 ml) 39 (630 mg, 63%) was
H,H
3
4
H, 2×3-H), 7.44 (dd, J
obtained as an amorphous orange coloured solid. MS (ESI,
H,H
H,H
2
+
2 H, 2 × 7-H), 4.13 – 4.08 (m, 4 H, 2 × CH (1’)), 4.05 (s,
4
5
.1 kV, 8 µl/min, N , methanol): 534 (18%) [MHK]
,
2
2
∗
2+
+
4 H, 2 × CH (2”)), 3.99 (s, 6 H, 2 × OCH ), 3.98 (s, 4 H,
58 (100%) [MH (NaCl)] , 1055 (15%) [MH] , 1094
2
3
2
+
35
35
∗
+
2 × CH (3”)), 3.51 (s, 4 H, 2 × CH (1”)), 3.25 – 3.20 (m,
(
[
(
7%) [MK] , 1112 [M( Cl)( Cl)H (NaCl)] , 1114
M( Cl)( Cl)H (NaCl)] , 1116 (9%) [M( Cl)( Cl)H
2
2
3
5
35
∗
+
37
35
∗
4 H, 2×CH2(12’)), 2.01 – 1.93 (m, 4 H, 2×CH2(2’)), 1.56 –
+
13
NaCl)] . – Analysis for C H Cl N O (1056.15): 1.22 (m, 36 H, 18 × CH2(3’-11’)). – C NMR (100 MHz,
56
72
2 8 8
C 63.69, H 6.87, N 10.61; found C 63.51, H 7.01, N 10.42.
CD3OD): δ = 170.3 (C=O), 167.9 (C=O), 158.1 (quart.),
1
1
57.8 (quart.), 141.8 (quart.), 130.2 (quart.), 128.6 (CH),
25.0 (CH), 121.4 (CH), 118.5 (CH), 116.1 (CH), 111.3
2
-((2-{[10-(9-{6-Chloro-2-methoxyacridinyl}amino)decyl]-
amino}-2-oxoethyl){2-[(2-{[10-(9-{6-chloro-2-methoxy-
acridinyl}amino) decyl]amino}-2-oxoethyl)(carboxymethyl)
amino]ethyl}amino) acetic acid (40)
(quart.), 57.6 (CH (3”)), 56.9 (OCH ), 56.0 (CH (2”)), 53.1
2 3 2
(CH (1”)), 50.4 (CH (1’)), 40.8 (CH (10’)), 30.79 (CH ),
2 2 2 2
3
0.68 (CH ), 30.61 (CH ), 30.57 (CH ), 30.42 (CH ), 30.26
2 2 2 2
(
CH ), 28.1 (CH ), 27.8 (CH ). – MS (ESI, 4.1 kV, 8 µl/min,
2 2 2
Following the procedure for 32 from 30 (1.5 g, 3.1 mmol)
3
+
3+
N , methanol): 381 (45%) [MH ] , 393 (24%) [MKH ]
,
2
3
2
and EDTA dianhydride (384 mg, 1.5 mmol) in DMF
2
+
2+
5
71 (100%) [MH ] , 590 (28%) [MHK] , 1140 (29%)
2
(30 ml) and triethylamine (5 ml) 40 (1.20 g, 74%) was ob-
3
5
35
+
37
35
+
[M( Cl)( Cl)H] , 1142 (48%) [M( Cl)( Cl)H] , 1144
tained as an amorphous orange coloured solid. MS (ESI,
3
7
37
+
+
2
+
(14%) [M( Cl)( Cl)H] , 1177 (10%) [MK] . – Analysis
for C H84 Cl2N8O8 (1116.29): C 64.56, H 7.59, N 10.04;
found C 64.37, H 7.69, N 9.86.
4
5
.1 kV, 8 µl/min, N , methanol): 543 (100%) [MH ]
,
2
2
2+
2+
6
2
63 (33%) [MHK] , 570 (27%) [MNaK] , 1084 (50%)
3
5
35
+
37
35
+
[M( Cl)( Cl)H] , 1086 (98%) [M( Cl)( Cl)H] , 1088
3
7
37
+
+
(20%) [M( Cl)( Cl)H] , 1121 (10%) [MK] . – Analysis
Acknowledgments
for C H Cl N O (1084.21): C 64.25, H 7.07, N 10.34;
5
8
76
2 8 8
found C 64.01, H 7.23, N 10.21.
Thanks for helpful discussion and parts of the biologi-
cal screening are due to Dr. H. Knoller and Dr. H.-J. Neu-
mann (Fresenius Hemocare); additional screening as been
performed by Bioscreen Ltd. Financial support by the Eu-
ropean Communities (SC1 -CT92-0780) and the Fonds der
Chemischen Industrie is gratefully acknowledged. We like to
2
-((2-{[12-(9-{6-Chloro-2-methoxyacridinyl}amino)-
dodecyl]amino}-2-oxoethyl){2-[(2-{[12-(9-{6-chloro-2-
methoxyacridinyl}amino) dodecyl]amino}-2-oxoethyl)-
∗
(carboxymethyl)amino]ethyl}amino) acetic acid (41)
Following the procedure for 32 from 31 (2.06 g, thank Dr. R. Kluge for the ESI-MS spectra and Dr. D. Str o¨ hl
4
.0 mmol) and EDTA dianhydride (512 mg, 2.0 mmol) for taking numerous NMR spectra.
[
[
[
[
1] H.-J. Neumann, H. Knoller, PCT WO 02/32875 A1;
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Unauthenticated
Download Date | 10/8/18 1:47 PM

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8
Ren e´ Csuk et al. · Synthesis of Dimeric Acridine Derived Nucleic Acid Intercalators
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Unauthenticated
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