Synthesis and antiviral activity of new bisacridinylhydrazides of aryloxyacetic acids

Full text of DOI:10.1023/A:1015336412246

Pharmaceutical Chemistry Journal
Vol. 35, No. 12, 2001
SYNTHESIS AND ANTIVIRAL ACTIVITY OF NEW
BISACRIDINYLHYDRAZIDES OF ARYLOXYACETIC ACIDS
S. A. Lyakhov,1 E. A. Lyakhova,1 N. N. Panchenko,2 L. A. Litvinova,1
and S. A. Andronati1
Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 35, No. 12, pp. 10 – 13, December, 2001.
Original article submitted September 14, 2000.
The class of aminoacridine derivatives contains com-
pounds possessing antitumor [1], antimicrobial [2], and anti-
viral [3] properties. As for acridinylhydrazine derivatives, it
was only established that some aryloxy acid acridinylhydra-
zides (I) exhibited fungicidal and schistosomiacidal activity
[4 – 8]. Previously [9], we reported that biodegradable
bisacridines (II, where X is a fragment containing amide or
ester bonds) may possess significant antiviral properties [9].
Purity of the target and intermediate compounds was
checked by TLC on Silufol UV-254 plates eluted in various
solvent systems. The compositions were confirmed by ele-
mental analyses; the proposed structures, by the data of IR
spectroscopy and mass spectrometry. The IR spectra exhib-
ited absorption bands characteristic of the intermolecularly
associated N–H bonds (3300 – 3250 cm – 1), aromatic and
aliphatic C–H bonds (3010 – 2930 cm – 1), ester and
hydrazide C=O groups (1710 – 1700 and 1650 – 1620 cm – 1
,
'
R
respectively), and double aromatic bonds (1500 –
1400 cm – 1). The mass spectra of all synthesized compounds
displayed the peaks due to molecular ions with intensities de-
creasing in the following order: diester > dihydrazide >
bisacridinylhydrazide. The yields and physicochemical char-
acteristics of the synthesized compounds are summarized in
Table 1.
R
O
N
N
O
X
HN
N
H
N
H
NH
II
N
The antiviral activity of the synthesized compound was
studied with respect to the influenza strain A2/Honkong
H1N1 (viral diagnosticum) in reference to amyxin [10]; the
tests were performed in ovo by conventional methods [11].
The results of these investigations are summarized in Ta-
ble 2.
I
In this context, we have synthesized and characterized a
series of bisacridinylhydrazides of dicarboxylic acids
XXV – XXXII. The target compounds were synthesized ac-
cording to the scheme. In the first step, 4,4¢-dihydroxy-
diphenyl (III), 2,2-bis(4-hydroxyphenyl)propane (IV), and
1,3- and 1,4-dihydroxybenzene (V, VI) of meta- and
para-hydroxybenzoic acid methylates (VII, VIII) were
alkylated by methylbromoacetate in boiling acetone in the
presence of potassium carbonate. The resulting diesters
IX – XIV, as well as methyl esters of isophthalic and
terephthalic acids (XV, XVI), were subjected to hydrazino-
lysis to obtain hydrazides XVII – XXIV. These hydrazides
were acridinylated by boiling with methoxyacridine in meth-
anol for 2 – 5 days. The target bisacridinylhydrazides XXV –
XXXII were obtained in a chromatographically pure state
with a yield of 75 – 85% directly from the reaction mixtures.
Some of the synthesized compounds (XXV – XXVII)
showed significant antiviral activity, as characterized by high
values (above 66%) of the protection index (PI). Some other
products were much less active (PI £ 50%), while two com-
pounds (XXX and XXXI) exhibited no antiviral effect at all
while being rather toxic (leading to the loss of one and three
test embryos in a group of six, respectively, in the absence of
any loss in the control group). Tested under analogous condi-
tions, amyxin was characterized by PI = 66.67%. Thus com-
pounds XXVI and XXVII exceeded amyxin in the antiviral
activity, while compound xXV was comparable with the ref-
erence drug in this respect. All tested substances led to a de-
crease in the viral titer, except for compound XXXI where
the decrease was not as reliable relative to control ( p > 0.05).
The obtained results confirm our previous data concern-
ing the antiviral activity of bisacridines [9]. Moreover, now
1
Bogatskii Institute of Physical Chemistry, National Academy of Sciences
of Ukraine, Odessa, Ukraine.
Odessa National University, Odessa, Ukraine.
2
653
0091-150X/01/3512-0653$25.00 © 2001 Plenum Publishing Corporation

654
S. A. Lyakhov et al.
Scheme
spectively (the average viral titer being
3.17 and 1.59, respectively).
O
O
O
O
A
A
Thus, an increase in the distance be-
tween acridine chromophores and in the
flexibility of a linker fragment enhances
the antiviral activity of compounds, this
rule being valid in the series of both
meta and para derivatives. The antiviral
activity significantly increases upon go-
ing from para derivatives with a single
aryl fragment to their analogs contain-
ing two benzene rings. The antiviral
properties of the latter compounds are
comparable with those of amyxin, while
the resorcinol derivative XXVII exceeds
the reference drug both in the PI value
and in the viral titer characteristic.
CH3
O
O
H3C
O
OH
OH
HO
IX
III
CH3
O
O
O
O
CH3
CH3
CH3
O
H3C
HO
CH3
X
O
IV
O
A
H3C
O
O
CH3
CH3
HO
OH
O
O
V
XI
A
O
O
O
O
H3C
O
OH
HO
O
XII
O
VI
Thus the synthesized bisacridines
are of interest for the further study as
potential antiviral agents.
A
O
O
O
H3C
OH
H3C
O
O
O
O
CH3
XIII
EXPERIMENTAL
CHEMICAL PART
VII
O
O
H3C
O
O
A
The mass spectra were recorded
with an MX-1321 instrument using a di-
rect sample introduction scheme. The
ion source was operated at an electron
impact energy of 70 eV and a tempera-
ture of 220°C. The IR spectra were mea-
sured in chloroform, carbon tetrachlo-
ride, or Vaseline oil in a Specord 75
spectrophotometer. The UV spectra
were recorded with Specord M-40 and
Perkin-Elmer Lambda 9 spectrophotom-
eters. TLC was performed on Silufol
UV-254 and UV-366 plates. The synthe-
ses were conducted with commercial re-
agents of not less than 97% purity rela-
tive to the main component.
O
O CH3
OH
H3C
XIV
VIII
H3C
O
O
O
O
H3C
H3C
CH3
O
O
CH3
O
O
O
XVI
XV
H
N
H
N
C
O
X
O
X
B
CH3
H2N
Acr
NH2
O
O
O
IX – XVI
XVII – XXIV
H
N
H
N
Acr
X
C
N
H
N
H
Acr =
O
O
N
XXV – XXXII
A: BrCH2COOCH3, K2CO3, (CH3)2CO, 55 °C, 8 h
B: N2H4 · H2O, CH3OH, 65 °C, 8 – 10 h
C: AcrOCH3, CH3OH, 65 °C, 50 – 120 h
1,4-Bis(carbomethoxy)methoxyben-
zene (XII).
A
mixture of 2.2 g
(0.02 mole)
hydroquinone, 6.9 g
(0.05 mole) potassium carbonate, and
3.97 ml (0.042 mole) of methylbromo-
we can establish certain relationships between the antiviral
properties and structure of these compounds. In particular,
the derivatives of isophthalic and terephthalic acids (XXXI,
XXXII) exhibit virtually no activity (PI = 0 and 16.67%, re-
spectively). Introduction of the oxymethylene fragment be-
tween carbonyl group and aromatic ring led to a decrease in
the viral titer and even to an increase in the PI value. The sec-
ond oxymethylene fragment introduced between the aro-
matic ring and another carbonyl increases the PI values of
compounds XXVIII and XXVII up to 16.67 and 83.33%, re-
acetate was boiled for 10 h with intensive stirring (prefera-
bly, under inert atmosphere), cooled to room temperature,
and filtered. The filtrate was evaporated under vacuum to
dryness and the dry residue was recrystallized from aqueous
methanol to obtain 3.32 g (65%) of compound XII.
Analogous procedures with phenols III – V, VII, and
VIII were used to obtain bis-esters IX – XI, XIII, and XIV,
respectively (Table 1).
2-[3-(Hydrazinocarbonylmethoxy)phenoxy]acetylhyd
razine (XIX). To a solution of 3.3 g (0.013 mole) of bis-ester

Synthesis and Antiviral Activity of New Bisacridinylhydrazides of Aryloxyacetic Acids
655
TABLE 1. Yields and Physicichemical Properties of the Synthesized Compounds
TLC mobility
Eluent**
Yield,
%
Molecular
weight.
Empirical
formula
Compound
M.p.,°C
Solvent
[M]+
Rf
IX
53
97
66
65
98
71
47
74
95
188 – 189
Dioxane
…
330.34
372.42
254.24
254.24
224.22
224.22
330.35
372.43
254.25
C18H18O6
C21H24O6
330
372
254
254
224
224
330
372
254
0.46
0.58
0.63
0.55
0.54
0.63
0.08
0.20
0.07
Chl–EA, 10 : 1
Chl–Ace, 5 : 1
Chl–Ace, 7 : 1
Chl–Ace, 7 : 1
Chl–Ace, 7 : 1
Chl–Ace, 7 : 1
Chl–Ace, 3 : 1
††
X
XI
62 – 63
MeOH–H2O
MeOH–H2O
…
C12H14O6
XII
98 – 99
††
C12H14O6
XIII
XIV
XVII
XVIII
XIX
C11H12O5
95 – 96
> 326 p†
191 – 192
246 – 247
MeOH–H2O
MeOH
C11H12O5
C16H18N4O4
C19H24N4O4
C10H14N4O4
MeOH
Chl–MeOH, 5 : 1
MeOH
BuOH–EtOH–H2O,
2 : 2 : 1
XX
93
262 – 263
MeOH
254.25
C10H14N4O4
254
0.04
MeOH–DMF–AcOH,
10 : 1 : 3
XXI
41
83
70
158 – 159
202 – 203
238 – 239
MeOH
MeOH
MeOH
224.22
224.22
194.19
C9H12N4O3
C9H12N4O3
C8H10N4O2
224
224
194
0.38
0.33
0.46
Chl–MeOH, 5 : 1
Chl–MeOH, 5 : 1
XXII
XXIII
MeOH–TEA–DMF,
20 : 1 : 2
XXIV
XXV
94
69
44
90
76
22
> 360 p
> 272 p
> 188 p
> 256 p
> 298 p
> 220 p
MeOH
MeOH
MeOH
MeOH
MeOH
MeOH
194.19
684.76
726.84
608.66
608.66
578.64
C8H10N4O2
C42H32N6O4
C45H38N6O4
C36H28N6O4
C36H28N6O4
C35H26N6O3
194
684
726
608
608
578
0.00
0.72
0.11
0.06
0.13
0.20
MeOH–TEA–DMF,
20 : 1 : 2
MeOH–TEA–DMF,
20 : 1 : 2
XXVI
XXVII
XXVIII
XXIX
Bnz–TEA–MeOH,
10 : 1 : 1
Bnz–TEA–MeOH,
10 : 1 : 1
Bnz–TEA–MeOH,
10 : 1 : 1
Bnz–TEA–MeOH,
10 : 1 : 1
XXX
41
78
84
224 – 225
> 362 p
MeOH
MeOH
MeOH
578.64
548.61
548.61
C35H26N6O3
C34H24N6O2
C34H24N6O2
578
548
548
0.56
0.51
Chl–MeOH, 5 : 1
MeOH–TEA, 20 : 1
MeOH–TEA, 20 : 1
XXXI
XXXII
> 374 p
0.00*
Notes: † (p), decomposition on melting; †† noncrystallizable oily product; * this substance was not eluted in any of the systems employed, while
all initial reagents and potential intermediate products were eluted to a certain extent; ** solvents: Ace = acetone, BuOH = butanol, Bnz = ben-
zene, DMF = dimethylformamide, MeOH-methanol, TEA = triethylamine, AcOH = acetic acid, Chl = chloroform, EtOH = ethanol,
EA = ethyl acetate.
XI in 20 ml of methanol was added 1.5 ml (0.03 mole) of
100% hydrazine hydrate and the mixture was boiled with
stirring for 8 h and cooled. The precipitate was separated by
filtration, washed on the filter with water (3 ´ 10 ml) and
50 h. The course of the reaction was monitored by TLC. Af-
ter terminating the process and cooling the mixture, the pre-
cipitate was separated by filtration, washed on the filter with
methanol (3 ´ 10 ml), and dried under vacuum
(10 – 15 Torr) at room temperature for 8 h to obtain 0.554 g
(90%) of compound XXVII.
Analogous procedures with bishydrazides XVII, XVIII,
and XX – XXIV were used to obtain compounds XXV,
XXVI, and XXVIII – XXXII, respectively (Table 1).
methanol (3 ´ 10 ml), and dried under vacuum
(10 – 15 Torr) at room temperature for 8 h to obtain 3.142 g
(95%) of compound XIX.
Analogous procedures with bis-esters IX, X, and
XII – XVI were used to obtain compounds XVII, XVIII, and
XX – XXIV, respectively (Table 1).
N-[3-(N¢-Acridin-9-yl-hydrazinocarbonylmethoxy]-
EXPERIMENTAL BIOLOGICAL PART
phenoxy)acetyl-(N¢-acridin-9-yl)hydrazine (XXVII). To a
suspension of 0.254 g (0.001 mole) of bishydrazide XIX in
10 ml of methanol was added 0.627 g (0.003 mole) of
methoxyacridine and the mixture was boiled with stirring for
The antiviral activity of the synthesized compounds was
studied by simultaneously introducing a sample and the test
viral species into allantochorion cavity of a 10-day chicken

656
S. A. Lyakhov et al.
TABLE 2. Antiviral Activity of Compounds XXV – XXXII
embryos were assigned a unity titer). The titration was per-
formed with respect to the developed hemagglutination reac-
tion with human erythrocytes of the zeroth group. The viral
titer was determined as a maximum dilution of the allantois
fluid for which erythrocyte agglutination was still observed
in the form of a film with scalloped lacy edge. The results of
titration were statistically processed relative to the control in
terms of the Wilcoxon criterion for independent data sets
[12].
Number of embryos
Compound
PC
PI
GMT
total
infected
%
XXV
6
6
6
6
6
5
3
6
6
6
2
1
1
5
3
5
3
5
2
6
33.33
16.67
16.67
83.33
50
3
6
66.67
1 : 2
XXVI
XXVII
XXVIII
XXIX
XXX
83.33 1 : 2.24
83.33 1 : 1.59
16.67 1 : 3.17
6
1.2
2
50
0
1 : 2.52
1 : 8
100
1
XXXI
XXXII
Amyxin
Control
100
1
0
1 : 10.1
1 : 16
1 : 2
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33.33
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DMSO to a concentration of 1.5 mg/ml and introduced in a
dose of 0.1 ml per embryo. The viral species were diluted
1 : 100 with DMSO and introduced into control and test em-
bryos in an amount of 0.1 ml. Each test and the control ex-
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embryos were incubated in a thermostat for 48 h.
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tection index (PI) calculated by the formula
PI = ((PC – 1)/PC) ´ 100%, where PC is the protection coef-
ficient determined as the ratio of the fraction of virus-in-
fected embryos in the control to that in the test group. For the
same PI value, a higher activity is assigned to a substance
characterized by a lower geometric mean titer (GMT) of the
virus.
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of titers for each embryos in a group of six (the virus-free
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