Reactions of aminoguanidine and guanidine with 3- and 5-formyl-4- arylaminopyridones

Article abstract of DOI:10.1007/s11172-010-0385-8

Amidinohydrazones were prepared by the condensation of 4-arylamino-2-oxo-5- formyl-1,2-dihydropyridine-3-carbonitriles and 4-arylamino-2-oxo-1,2- dihydropyridine-3-carbaldehydes with aminoguanidinium carbonate with the purpose to investigate their biological activity as putative NO donors. The reaction of 5- Ee Cyrillic sign 3-formylpyridones with diguanidinium carbonate was studied. The formation of stable complexes of 4-arylamino-5-formyl-2-oxo-1,2- dihydropyridine-3-carbonitriles with aminoguanidine and guanidine was discovered. Amidinohydrazones obtained possess antiinflammatory, antidiabetic, and antihypertensive activities.

Full text of DOI:10.1007/s11172-010-0385-8

Russian Chemical Bulletin, International Edition, Vol. 59, No. 12, pp. 2247—2258, December, 2010
2247
Reactions of aminoguanidine and guanidine
with 3ꢀ and 5ꢀformylꢀ4ꢀarylaminopyridones
M. I. Medvedeva,a N. Z. Tugusheva,b L. M. Alekseeva,b M. A. Kalinkina,b V. A. Parshin,b V. V. Chernyshev,c
V. I. Levina,b N. B. Grigor´ev,b A. S. Shashkov,d and V. G. Granikb
aD. I. Mendeleev University of Chemical Technology of Russia,
9 pl. Miusskaya, 125047 Moscow, Russian Federation.
Eꢀmail: marinacasper@gmail.com
bMoscow Theoretical and Practical Center for Narcology,
37/1 ul. Lyublinskaya, 109390 Moscow, Russian Federation.
Eꢀmail: vggranik@mail.ru
cDepartment of Chemistry, M. V. Lomonosov Moscow State University,
Build. 3, 1, Leninskie Gory, 119992 Moscow, Russian Federation.
Eꢀmail: cher@biocryst.phys.msu.su
dN. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences,
47 Leninsky prosp., 119991 Moscow, Russian Federation.
Fax: +7 (499)135 5328. Eꢀmail: shash@ioc.ac.ru
Amidinohydrazones were prepared by the condensation of 4ꢀarylaminoꢀ2ꢀoxoꢀ5ꢀformylꢀ
1,2ꢀdihydropyridineꢀ3ꢀcarbonitriles and 4ꢀarylaminoꢀ2ꢀoxoꢀ1,2ꢀdihydropyridineꢀ3ꢀcarbꢀ
aldehydes with aminoguanidinium carbonate with the purpose to investigate their biological
activity as putative NO donors. The reaction of 5ꢀ и 3ꢀformylpyridones with diguanidinium
carbonate was studied. The formation of stable complexes of 4ꢀarylaminoꢀ5ꢀformylꢀ2ꢀoxoꢀ1,2ꢀ
dihydropyridineꢀ3ꢀcarbonitriles with aminoguanidine and guanidine was discovered. Amidinoꢀ
hydrazones obtained possess antiinflammatory, antidiabetic, and antihypertensive activities.
Key words: 4ꢀarylaminoꢀ5ꢀformylꢀ2ꢀoxoꢀ1,2ꢀdihydropyridineꢀ3ꢀcarbonitriles, 4ꢀarylamiꢀ
noꢀ2ꢀoxoꢀ1,2ꢀdihydropyridineꢀ3ꢀcarbaldehydes, complexes of 4ꢀarylaminoꢀ5ꢀformylꢀ2ꢀoxoꢀ
1,2ꢀdihydropyridineꢀ3ꢀcarbonitriles with aminoguanidine and guanidine, 2ꢀoxoꢀ4ꢀanilinoꢀ1,2ꢀ
dihydropyridineꢀ3ꢀcarbonitrile, dimethylformamide dimethylacetal, aminoguanidine, guaniꢀ
dine, Xꢀray diffraction, antiinflammatory, antidiabetic, and antihypertensive activities.
It is well known that nitric oxide (NO) plays a key role
in the control of vascular tone, in the maintenance of
cardiovascular homeostasis, in the control of breathing,
immunity, neurotransmission mechanisms, and at the
same time it is a cytotoxic and cytostatic agent. Quite
a number of pathological states, e.g., cardiovascular, inꢀ
fectious, inflammatory, and other diseases, can be related
to the lack or overproduction of NO in the organism.1—8
In this connection, one of the most actively developing
line of investigations is currently the search of various
compounds which are able to serve as generators or inhibꢀ
itors of NO formation, i.e., the search of compounds,
whose transformation can lead to formation of NO in the
organism or inhibition of its production under physioꢀ
logical conditions.
volves both the amino acid and, first of all, guanidine
fragment (at the special site of «guanidine binding» and
interaction with heme).9 Hence, it was concluded that
compounds containing guanidine or guanidineꢀlike groups
can be the substrates of NO synthases (NOS) and catalytꢀ
ically transformed with the release of NO. Furthemore, it
is known that such compounds as aminoguanidine are
selective inhibitors of inducible NO synthase, which acꢀ
counts for considerable biological activity of this comꢀ
pound or its derivatives under various pathologies.1
In the present work, a number of guanidine and amiꢀ
noguanidine derivatives of 4ꢀaminoꢀ1,2ꢀdihydropyridinꢀ
2ꢀone was synthesized and some of their parameters of
antiinflammatory, antidiabetic, and antihypertensive acꢀ
tivities were investigated.
It is known that the guanidine fragment of Lꢀarginine
being a target for NO synthases undergoes oxidation to the
Nꢀhydroxy derivative and then to citrulline and nitric
oxide.1 Binding of substrates to enzyme isoforms inꢀ
Earlier,10 we have prepared 4ꢀarylaminoꢀ1,2ꢀdihydroꢀ
pyridinꢀ2ꢀones containing a formyl group in positions 3 or
5. Using HPLC, it was established that 5ꢀformylpyridones
are much more reactive in reactions with carbon acids
Published in Russian in Izvestiya Akademii Nauk. Seriya Khimicheskaya, No. 12, pp. 2191—2202, December, 2010.
1066ꢀ5285/10/5912ꢀ2247 © 2010 Springer Science+Business Media, Inc.

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Russ.Chem.Bull., Int.Ed., Vol. 59, No. 12, December, 2010
Medvedeva et al.
than 3ꢀformylpyridones. It was explained by probable steric
hindrance in the first step of the condensation.11 In conꢀ
tinuation of our studies on comparison of the reactivity of
the 3ꢀ and 5ꢀformyl groups in 4ꢀarylaminopyridones, we
describe here reactions of 4ꢀarylaminoꢀ5ꢀformylꢀ2ꢀoxoꢀ
1,2ꢀdihydropyridineꢀ3ꢀcarbonitriles (1а,b) and 4ꢀarylamiꢀ
noꢀ2ꢀoxoꢀ1,2ꢀdihydropyridineꢀ3ꢀcarbaldehydes (2а,b)
with aminoguanidine and guanidine.
essential that their structures were established by both specꢀ
troscopic and Xꢀray diffraction analysis.
The appearance of the precipitate changed upon longꢀ
term refluxing of compounds 5а,b (24 h for 5а and 32 h
for 5b), the isolated products are the expected coupling
products 3а,b as the free bases. The structure of compound
3а was established by HMBC NMR spectroscopy (see
Experimental) and powder Xꢀray diffraction analysis data.
The reaction of aminoguanidinium hydrogencarbonꢀ
ate in 96% ethanol with both 5ꢀformylpyridones 1а,b and
3ꢀformylpyridones 2а,b in the presence of an acid results
in the expected coupling products 3а,b and 4a,b isolated
as hydrochlorides in good yields (Schemes 1 and 2). If
these reactions are carried out in pyridine in the absence of
the acid, the reactions of 5ꢀ and 3ꢀformylpyridones follow
different routes. White voluminous precipitates were
formed upon refluxing of 5ꢀformylpyridones 1а,b with
a twofold excess of aminoguanidinium hydrogencarbonꢀ
ate for 10—30 min. The IR spectra of compounds 5а,b
obtained show signals for the NН, NН2, СN, СНО, and
СО groups (see Experimental). The 1Н NMR spectra conꢀ
tain, in addition to the signals corresponding to the strucꢀ
ture of the starting 5ꢀformylpyridones but shifted upfield
by 0.3 ppm, two broadened signals at δ 4.67 (2 Н) and
δ 7.30 (4 Н) for 5а, or δ 4.63 (2 Н) and δ 7.24 (4 Н) for 5b,
which can be assigned to the signals for the NH2 and
NН groups of aminoguanidine. Compound 5а was invesꢀ
tigated in more detail. The overlap of the signals for the
protons of CHO, H(6) and phenyl ring was observed in
the 1Н NMR spectrum of a mixture of the starting
Scheme 1
1
5ꢀformylpyridone 1а and compound 5а, while Н NMR
spectrum of a mixture of compounds 3а as a free base and
5а shows double set of the signals corresponding to the
individual compounds 3а and 5а. In the 13С NMR spectra
of compounds 1а and 3а, the chemical shifts of carbonyl
atom С(2)=О virtually coincided (δ 161.4 and 161.5),
while in the spectrum of compound 5а it was shifted
downfield (δ 172.9), which can be explained by formaꢀ
tion of a hydrogen bond with the H—N group of aminoꢀ
guanidine. The assignment of the signals in the 13С NMR
spectra of compounds 3а and 5а was made based on corꢀ
relation peaks in 2D HMBC spectrum (see Experimenꢀ
tal). The 13С NMR spectrum of compound 5а shows
the signal at δ 159.0, which can be assigned to the C atom
of the aminoguanidine fragment, c.f. the chemical
shift of NНC(=NH) in the 13С NMR spectrum of comꢀ
pound 3а, δ 159.0. According to these data and data
from elemental analysis for 5а, the formation of compꢀ
lexes under shortꢀterm refluxing of aminoguanidinium
hydrogencarbonate with 5ꢀformylpyridones 1а,b in pyriꢀ
dine was supposed. This was also confirmed by powder
Xꢀray diffraction analysis of compound 5а (see Exꢀ
perimental).
R = H (a), Cl (b)
Reagents and conditions: i. Aminoguanidinium hydrogencarbonꢀ
ate, Py, 24 h or aminoguanidinium hydrogencarbonate, EtOH,
HCl, H2O; ii. Aminoguanidinium hydrogencarbonate, Py,
30 min; iii. Longꢀterm refluxing in pyridine.
Coupling of aminoguanidine with 3ꢀformylpyridones
2а,b in pyridine proceeds much faster (5 h for 2а, 15 h
for 2b). The expected amidinohydrazones 4а,b were
isolated in good yields (Scheme 2).
The reaction of 5ꢀformylpyridone 1а with diguanidiniꢀ
um carbonate in pyridine (Scheme 3) also results in comꢀ
plex 6 analogous to complexes 5а,b (the rection time was
30 min), its structure was confirmed by the data from
1Н NMR, IR, and mass spectra and powder Xꢀray diffracꢀ
tion analysis (see Experimental).
All geometrical characteristics of the molecules in the
crystal structures of 3а, 5а, 6 (Fig. 1) have values similar
to those found in Cambridge Crystallographic Database
(CCDC).12 The crystal packings in all compounds are
characterized by the presence of classic intermolecular
hydrogen bonds N—H...N and N—H...O (Table 1). The
presence of centrosymmetrical tetramers uniting two main
It should be noted that the formation of such complexꢀ
es is an interesting and unusual fact, and in this case it is

Reactions of aminoguanidine and guanidine
Russ.Chem.Bull., Int.Ed., Vol. 59, No. 12, December, 2010
2249
Scheme 2
N(21)
C(20)
N(19)
C(6)
C(5)
N(1)
C(2)
C(17)
N(18)
C(4)
N(22)
C(3)
C(8)
O(7)
N(10)
C(11)
C(4)
C(16)
C(12)
C(13)
N(9)
C(15)
C(14)
3a
C(6)
O(7) N(1)
N(22)
N(21)
C(10)
C(5)
C(2)
C(3)
C(8)
C(19)
N(20)
C(4)
N(12)
R = H (a), Cl (b)
O(11)
Reagents and conditions: i. Aminoguanidinium hydrogencarbonꢀ
ate, Py; ii. Aminoguanidinium hydrogencarbonate, 96% EtOH,
HCl, H2O.
N(9)
C(18)
C(17)
C(13)
C(14)
Scheme 3
C(16)
C(15)
6
O(11)
C(10)
C(6)
C(5)
N(23)
N(1)
N(12)
N(21)
C(19)
C(18)
C(4)
C(17)
C(13)
C(2)
O(7)
N(22)
C(3)
C(8)
C(14)
C(15)
N(20)
C(16)
Conditions: Py, 30 min.
N(9)
5a
molecules and two molecules of guanidines bound by hyꢀ
drogen bonds can be related as an interesting feature of
packing of 6 and 5a (Fig. 2).
Fig. 1. Molecular structure of compounds 3a, 6, and 5a. Spheres
of atom displacements are presented with 50% probability for
nonhydrogen atoms.
Only the starting 5ꢀformylpyridone 1а was isolated
in quantitative yield under carrying out the reaction
of diguanidinium carbonate with 5ꢀformylpyridone in
96% ethanol in the presence of an acid regardless of the
reaction time (3.5 h or 35 h), and also upon refluxꢀ
ing complex 6 under the same conditions (the rection
time was 11 h).
The reaction of 3ꢀformylpyridone 2а with diguanidiꢀ
nium carbonate occurs differently. Deformylation prodꢀ
uct, viz. 4ꢀanilinopyridone 7 (Scheme 4), was dominant
under longꢀterm refluxing of 3ꢀformylpyridone 2а in pyriꢀ
dine (22 h), it was also prepared by us from 4ꢀanilinoꢀ2ꢀ

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Russ.Chem.Bull., Int.Ed., Vol. 59, No. 12, December, 2010
Medvedeva et al.
Table 1. Characteristics of hydrogen bonds in compounds 3а, 5а, and 6
Comꢀ
pound
D—H...A
D—H
H...A
D...A
D—H...A
/deg
Å
3а
N(1)—H(1)...O(7)i
0.86
0.86
0.86
0.86
0.86
1.82
2.13
2.56
2.33
2.20
2.676(10)
2.728(10)
3.411(11)
2.930(11)
2.747(10)
171
127
171
127
122
N(10)—H(10)...N(18)
N(21)—H(21A)...N(9)ii
N(21)—H(21B)...N(22)iii
N(21)—H(21B)...O(7)iv
5а
N(1)—H(1)...N(21)
0.86
0.86
0.86
0.86
0.86
0.86
0.90
2.07
2.02
1.89
2.18
2.28
2.13
2.47
2.894(19)
2.69(2)
2.743(19)
2.918(18)
2.611(18)
2.99(2)
161
134
170
144
103
175
131
N(12)—H(12)...O(11)
N(20)—H(20A)...O(7)
N(20)—H(20B)...O(7)vii
N(22)—H(22A)...N(23)
N(22)—H(22B)...N(9)vii
N(23)—H(23A)...O(11)viii
3.143(18)
6
N(1)—H(1)...N(21)v
N(12)—H(12)...O(11)
N(20)—H(20A)...O(11)vi
N(20)—H(20B)...N(9)
N(21)—H(21)...O(11)vi
N(22)—H(22A)...O(7)v
N(22)—H(22B)...O(7)
0.86
0.86
0.86
0.86
0.86
0.86
0.86
2.02
2.00
2.30
2.17
2.30
1.86
2.16
2.872(12)
2.714(12)
3.058(13)
3.009(14)
3.066(12)
2.713(12)
2.869(13)
172
139
147
164
148
175
139
Note. Code of symmetry: (i) –x, y – 1/2, 1/2 – z; (ii) –x, 1 – y, –z; (iii) x, y – 1, z;
(iv) x, –y – 1/2, z – 1/2; (v) –x, 1 – y, 1 – z; (vi) x – 2, 1/2 – y, z – 1/2; (vii) 1 – x, 2 – y, z;
(viii) –1 – x, 1 – y, z.
oxoꢀ1,2ꢀdihydropyridineꢀ3ꢀcarbonitrile 8 by hydrolysis of
the cyano group to the carboxyl group and subsequent
decarboxylation of the latter.
a
Scheme 4
b
Reagents and conditions: i. Diguanidinium carbonate, Py, 22 h
or EtOH, HCl, H2O; ii. KOH, ethylene glycol.
Deformylated pyridone 7 is the dominant product too
under carrying out the reaction of 3ꢀformylpyridone 2а
with guanidine carbonate in 96% ethanol in the presence
of the acid (the reaction time was 20 h).
Next we studied the reactions of the condensation
products obtained 3а,b and 4а with dimethylformamide
dimethylacetal. Bisꢀamidine 9а (Scheme 5) formed in the
reaction of 5ꢀsubstituted pyridone 3а as both the free base
and hydrochloride with Me2NCH(OMe)2 under mild conꢀ
ditions (keeping at 20 °С in toluene or in isopropyl alcoꢀ
hol). This compound is unstable and decomposed in atꢀ
tempted recrystallization, therefore we failed to isolate
Fig. 2. Centrosymmetrical tetramers in the crystal structure
of 6 (a) and 5a (b) formed due to intermolecular hydrogen
bonds N—H...N and N—H...O (interaction is indicated by
dashed lines).

Reactions of aminoguanidine and guanidine
Russ.Chem.Bull., Int.Ed., Vol. 59, No. 12, December, 2010
2251
Table 2. The ability of compounds 3а,b, 4a,
and 9b,c to release of NO
compound isolated after refluxing in toluene suggests the
presence of traces of a methylation product. Compound 10
like 9а, is thermally unstable, and the appearance of
1,2ꢀdihydrobenzo[b]ꢀ[1,6]ꢀnaphthyridinꢀ2ꢀone (obtained
from 1b by us earlier10) in the sample was observed.
Compound
The yield of NO2– (%)
3a•HCl
3b
4a
9b
7.7 0.8
8.8 0.8
5.8 0.68
4.2 0.4
3.9 0.4
10.2 1.0
Scheme 6
9c
Guanabenz
it in pure form. Its structure was established by data from
1
mass spectrometry and H NMR spectroscopy (see Exꢀ
perimental). In addition to condensation, alkylation of
the pyridine nitrogen atom occurs when conducting the
reactions of 3а,b with Me2NCH(OMe)2 under more drasꢀ
tic conditions (refluxing in toluene), and as a result
analytically pure products 9b,c were isolated in satisꢀ
factory yields.
As was mentioned above, the aim of the present study
was the synthesis of a group of new compounds containing
the guanidine fragment. This, in turn, was caused by caꢀ
pability of oxidation of these compounds in the organism
with release of nitric oxide. Therefore, it was of interest to
study the biological activity of the compounds obtained
with regard to parameters typical of NO donors.
Scheme 5
First, the ability of compounds 3а,b, 4b, and 9b,c to
release NO was studied. The results from Table 2 demꢀ
onstrate these compounds to be NO donors.
The ability of compounds 3а,b, 4b, and 9b,c to release
NO upon oxidation was determined in their chemical
oxidation with a solution of mꢀchloroperbenzoic acid
(рH 9.8) (20ꢀfold excess) followed by passing the solution
through a column with porous cadmium.
The results obtained were compared with the ability of
a hypotensive medicine Guanabenz to release NO under
identical conditions. Guanabenz was shown13 to undergo
transformation into nitric oxide under incubation with
cytochrome oxidase of liver microsomes and therefore, it
can be the exogenous source of NO in the organism.
As a result of biological tests in normotensive and hyꢀ
pertensive rats with ultimate arteriotony corresponding to
180 12.5 Torr, compound 4а in the dose of 10 mg kg–1
was shown to decrease arteriotony. Compound 4а is comꢀ
R = R´ = H (a); R = H, R´ = Me (b); R = Cl, R´ = Me (c)
3ꢀSubstituted pyridone 4а in contrast to 5ꢀsubstituted
pyridones 3а,b is less susceptible to alkylation by the aceꢀ
tal. The reaction of 4а as both the free base and hydroꢀ
chloride with Me2NCH(OMe)2 results in amidine 10 unꢀ
der both mild conditions (keeping at 20 °С in toluene or in
isopropyl alcohol) and more drastic conditions (refluxing
in toluene for 6 h) (Scheme 6). The mass spectrum of the
Table 3. The influence of compound 4а under studying upon the system arteriotony for rats
Compound
Dose/mg kg–1
(intravenously)
Reduction of arteriotony (Δ) relatively to the initial level
Δ/Torr (%)
Normotensive
Hypertensive
rats (n = 5)
rats (n = 5)
4а
10
10
15.0 0.8 (10.7)
19.5 0.7 (13.9)
17.0 2.0 (9.4)
22.5 0.5 (12.5)
5ꢀMononitrate
Isosorbide

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Medvedeva et al.
Table 4. The influence of compounds 3а•HCl, 3b•HCl, 4а, and 9b upon pain reaction of mices caused by
intraperitoneal introduction of the solution of acetic acid
Compound
The number of
animals
Dose/mg kg–1 The number of writhes* for mices
Analgesic
activity**
(perorally)
m (%)
Control
3a•HCl
30
10
10
10
10
10
10
10
10
10
19.8 2.6 (100)
18.8 1.1 (95.0)
17.5 0.8 (88.4)
17.3 2.0 (87.4)
16.1 1.5 (81.3)
17.9 1.4 (90.4)
17.0 1.45 (85.6)
18.9 1.2 (95.5)
18.0 1.4 (91.1)
05.3 0.5 (26.8)
100
200
100
200
100
200
100
200
5.0
5.0
11.6
12.6
18.7
9.6
14.4
4.5
8.9
3b•HCl
4a
9b
Indometacin
73.2
* Pain reaction upon abdominal membrane sensation. ** Relative to the control (%).
pared with isosorbideꢀ5ꢀmononitrate in the same dose in
terms of the hypotensive effect (Table 3).
Thus, the investigation of pharmacological activity of
guanidine derivatives, viz., compounds 3а•HCl, 3b•HCl,
4а, and 9b as regards their antihypertensive, antiinflamꢀ
matory, analgesic, and hypoglycemic activities recognized
all compounds to possess weak antiinflammatory activity.
Compound 4а possessing antiinflammatory, hypoglyꢀ
cemic, and antihypertensive activities and compound
3а•HCl possessing antiinflammatory and antihypertenꢀ
sive activities have the most significant spectrum of activꢀ
ity. Their activity is weaker in comparison with known
drugs.
The analogous spectrum of activity is noted for the NO
donor aminoguanidine (antiinflammatory, hypoglycemꢀ
ic, and hypertensive activities), which is used in the treatꢀ
ment of diabetic retinopathies and nephropathies. Amiꢀ
noguanidine increases arteriotony (probably, it is imporꢀ
tant for the treatment of microangiopathies), while comꢀ
pound 4а decreases arteriotony, which could be promising
for the development of new peripheral vasorelaxants in
this class of compounds.
Compound 4а, as well as 3а•HCl, 3b•HCl, and 9b in
doses 100 and 200 mg•kg–1 with oral administration posꢀ
sesses slight analgesic effect in mice in the test of writhes
caused by intraperitoneal injection of a solution of acetic
acid (Table 4).
Compounds 3а•HCl, 3b•HCl, 4а, and 9b in doses 100
and 200 mg kg–1 under oral administration possess antiinꢀ
flammatory action in mice on the peritonitis models proꢀ
duced by LPS and carrageenan. This effect of compounds
under study is much less expressed than that of indometaꢀ
cin (Table 5).
Compound 4а in doses 100 and 400 mg kg–1 possesses
hypoglycemic action on the model of srteptozocin diabeꢀ
tes in mice, but it yield in activity of metformin (Table 6).
In studies of overall toxic properties, compounds
3а•HCl, 3b•HCl, 4а, and 9b were shown to be low toxic
compounds, LD50 for these compounds is more than
1000 mg kg–1 under oral administration.
Table 5. Antiinflammatory activity of compounds 3а·HCl, 3b·HCl, 4а, and 9b on the peritonitis models
produced by introduction of lipopolysaccharide (LPS) for mices
Compound
Dose/mg kg–1
(perorally)
LPSꢀperitonitis
Carrageenanꢀperitonitis
V/mLa
Inflammation (%)
V/mLa
Inflammation (%)
3а•HCl
3b•HCl
4а
100
200
100
200
100
200
100
200
5.0
3.5 0.35
2.85 0.45b
3.5 0.5
2.9 0.45b
3.15 0.35
2.65 0.40b
3.7 0.3
3.0 0.3
1.0 0.12
3.8 0.5
7.9
25.0
7.9
23.7
17.1
30.3
2.6
21.1
73.7
0
0.4 0.1
3.4 0.5
88.2
0
2.8 0.35b
2.8 0.35b
2.45 0.3b
2.7 0.3b
2.45 0.3b
2.7 0.3b
2.8 0.35b
2.8 0.35b
17.6
17.7
27.9
20.6
27.9
20.6
17.6
17.7
9b
Indometacin
Control (LPS)
a V — the volume (size) of the substrate. *Р < 0.05 under comparision with control.

Reactions of aminoguanidine and guanidine
Russ.Chem.Bull., Int.Ed., Vol. 59, No. 12, December, 2010
2253
Table 6. Intensity and continuance of hypoglycemic activity of compounds 3а•HCl, 3b•HCl, 4а, and 9b
Compound
Dose/mg•kg–1
(perorally)
The mean level of glucose
in mices´ blood/mol L–1
Reduction of the level
glucose to control 2
Sugarꢀlowering
effect, N/h
Δ
M (%)
Initial level
After compounds
3а•HCl
3b•HCl
4а
400.0
400.0
100.0
400.0
400.0
100
200
400
—
14.5 1.2
14.0 1.3*
14.4 1.5*
14.3 1.5*
13.5 1.2
13.7 1.4
13.8 1.5
14.7 1.4
6.3 1.0
14.4 1.4
14.0 1.5
12.0 1.5**
11.1 1.0**
13.4 1.3
8.4 1.4**
7.7 1.0**
7.1 1.5**
—
Non active
Non active
16.7
23.1
Non active
38.7
—
—
2.0
2.0
—
3.0
3.0
4.0
—
9b
Metformin
44.2
51.7
100
Control 1
(mices without diabete)
Control 2
—
14.3 1.5
—
0
—
(mices with diabete)
* Differences with control I at P < 0.05. ** Differences with control II at P < 0.05.
(2 : 1 : 7, 4 volumes). The solutions obtained were passed through
columns with porous cadmium prepared according to the same
State Standard Specification. The content of the NO2– anion in
the effluents was determined by the Griess reaction (diazo comꢀ
ponent is sulfanilic acid, azo component is 1ꢀnaphthylethyleneꢀ
diamine). It should be noted that the nitrite anion is an accepted
marker of nitric oxide. The determination was carried out in
0.1 М hydrochloric acid by the method of additions. The accuꢀ
racy of determination was 10%.
Furthermore, the combination of three various types
of activity for compound 4а (antiinflammatory, hypoglyꢀ
cemic, and antihypertensive activities) can be promising
for the treatment of diabetic angiopathies (angiopathy
is injury of the blood vessels of various size, from capilꢀ
laries to large vessels, caused by the nervous regulation
failure), when not only vascular and metabolic comꢀ
ponents, but elements of inflammatory process are also
involved.
Biological studies
Experimental
1. The influence upon the arteriotony for rats. The investigations
were carried out using pedigreeless ratꢀmales (250—270 g each)
which were normotensive with initial arteriotony of 40 25 Torr
and hypertensive animals.14
Hypertension was produced by inhibitor of NO synthase
(nitroꢀLꢀarginine). Inhibitor of NO synthase was injected to the
rat's stomach in the form of 1% solution during 21 days on the
basis of 3.0 mg per one animal daily. In 3 weeks stable increasing
of the arteriotony to 180—185 Torr was became.
System arteriotony was monitored in the left carotid of the
animals narcotized by urethane (1.5 g kg–1 intraperitoneally) by
means of ADInstruments complex (Australia) consisted of the
pressure sensor joined with signal reduction system and displayꢀ
ing on computer monitor by means of Chart v. 4.2.3 program.
The compounds under studying were dissolved in 20—50% ethaꢀ
nol and injected into jugular vein via catheter. Each compound
was tested on 3—4 animals.
2. Hypoglycemic activity. Hypoglycemic activity was investiꢀ
gated using white pedigreeless miceꢀmales (24—25 g each) with
experimental pancreatic diabete.15
For the simulation of pancreatic diabete of the second type
(nonꢀinsulinꢀdependent type of pancreatic diabete) streptozocin
was injected intraperitoneally in dose 30.0 mg kg–1 during three
days. Blood for glucose test was taken from tail vein, glucose
testing was carried out by means of test strips of blood glucose
meter AccuꢀChek Active (Germany).
The IRꢀspectra were recorded on an FSMꢀ1201 instrument
in Nujol mulls. The electrospray ionization (ESI) mass spectra
were recorded on a Waters ZQꢀ2000 mass spectrometer using
a direct inlet system without a chromatographic column. The
1H NMR spectra were recorded on a Bruker ACꢀ300 spectromeꢀ
ter in DMSOꢀd6 and DMSOꢀd6—CCl4. The 13C NMR spectra
were recorded on a Varian Unity+400 (operating frequency
100 MHz) in DMSOꢀd6. The numeration shown in the Schemes
was used for the description of the NMR spectra. The course of
the reactions was monitored, and the purity of the compounds
was checked, by TLC on Silica gel 60 F254 plates (Merсk) (chloꢀ
roform; chloroform—ethanol, 10 : 1; ethyl acetate—ethanol
10 : 1; acetone). The melting points were determined on an
Electrotermal 9100 instrument (UK). N,Nꢀdimethylformamide
dimethylacetal (purity 97%, Lancaster) was used.
Examination of the ability of compounds 3а,b, 4a, and 9b,c to
release NO upon oxidation (general procedure). Exact amounts
(5•10–6 mol) of compounds 3а,b, 4a, and 9b,c were dissolved in
0.02 М solution of mꢀchloroperbenzoic acid (5 mL) in a 1 : 1
mixture of 96% ethanol and aqueous borate buffer solution
(pH 9.8). The solutions obtained were kept at 27 1 °С for 2 days.
Then aliquots withdrawn from the solutions were diluted with
a buffer solution made of ammonia buffer solution (pH 9.6, State
Standard Specification 29270ꢀ95, Products of processing of fruits
and vegetables. Methods for nitrate test), 96% ethanol and water

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Russ.Chem.Bull., Int.Ed., Vol. 59, No. 12, December, 2010
Medvedeva et al.
The compounds under investigations were tested orally in
doses 100.0 and 400.0 mg•kg–1. At present work metformin was
the medicine of comparision in doses 200.0—400.0 mg kg–1
(orally).
3. Analgesic activity. Analgesic activity of compounds was
investigated using miceꢀmales (20—22 g each, in groups of
10 animals) under chemical painful stimulation.16
Painful stimulation was caused by intraperitoneal injection
of 0.75% acetic acid (0.1 mL•10 g–1 of body weight). During
15 min the number of writhes (it is a pain reaction caused by
irritation of peritoneum) was counted for the animals placed into
individual cage. The compounds under studying in 100 and
200 mg kg–1 doses were injected into the stomach by pump an
hour before injection of acetic acid. Indometacin in 5.0 mg kg–1
(orally) was the medicine of comparision.
4. Antiinflammatory activity. Antiinflammatory activity was
estimated on mices by antiexudative action on the on the peritoꢀ
nitis models produced by lipopolysaccharide (LPS)17 and carraꢀ
geenan.18
Experiments were carried out using miceꢀmales (22.0—23.0 g
each, in groups of eight animals). Peritonitis was caused by intraꢀ
peritoneal injection of lipopolysaccharide (LPS) isolated from
Escherichia coli (Sigma) in 1.0 mg•kg–1 or 0.2 mL of 1%
Xꢀcarrageenan.
In 4 h after injection of LPS animals were slaughtered (by
means of CO inhalation), abdominal cavity was dissected and
exudate volume (mL) was measured. Compounds under investiꢀ
gation were injected orally (per os) in 100 and 200 mg kg–1
doses an hour before LPS. The control animals were given
0.3 mL of physiologic saline orally. Indometacin in 5.0 mg kg–1
(orally) was the medicine of comparision.
The results of the experiments were processed by the methꢀ
ods of variational statistics for biological investigations (detecꢀ
tion of arithmetic average, standard error, Student criterion t
was applyed under comparison of average).
5. The determination of the acute toxicity of the compounds
investigated under single dose for mice. The experiments were
carried out using miceꢀmales (18—20 g each). The compounds
under studying were injected as a suspension in water with twinꢀ80
into stomach. Each dose was injected to five animals. Behaviour
and state of health was looked at during 5 days. LDзо was estiꢀ
mated acording to the Kerber's method.19
ethanol (4 mL) stirred under reflux. The reaction mixture was
stirred under reflux for 7 h (the precipitate was not dissolved
under these conditions), and cooled to 20 °С. The precipitate
was filtered off and washed with 96% ethanol. Compounds 3а,b
and 4а,b were obtained as hydrochlorides.
2ꢀ[(4ꢀAnilinoꢀ3ꢀcyanoꢀ2ꢀoxoꢀ1,2ꢀdihydropyridinꢀ5ꢀyl)ꢀ
methylidene]aminoguanidine (3а) was prepared from compound
1а according to the method А. M.p. 298—300 °С (DMF : PriOH,
1 : 1), the yield was 78%. ESI MS, m/z: 296 [М + Н]+, 591
[2 М + Н]+. 1Н NMR (DMSOꢀd6), δ: 5.49, 5.66 (both br.s, 2 Н
each, NНC(=NH)NH2); 7.22 (t, 1 Н, Ph, Jо = 6.9 Hz);
7.30—7.41 (m, 4 Н, Ph); 7.74 (s, 1 Н, Н(6)); 8.04 (s, 1 Н,
C(5)CН); 11.58 (s, 1 Н, С(4)NН). 13С NMR (DMSOꢀd6), δ:
79.5 (С(3)); 106.05 (С(5)); 115(С≡N); 124.7, 125.9, 128.6,
137.6 (Ph); 141.0 (С(6)); 144.4 (C(5)CН); 155.3 (С(4)); 159.0
(NНC(=NH)); 161.5 (С(2)). The assignment of the signals in
the 13С NMR spectrum of compound 3а was made based on the
HMBC technique. Characteristic correlation peaks in the HMBC
spectrum of compound 3a are as follows: 7.74/106.0 Н(6)/C(5),
7.74/144.4 Н(6)/C(5)CН, 7.74/155.3 Н(6)/C(4), 7.74/161.5
Н(6)/C(2), 8.04/106.0 C(5)CН/C(5), 8.04/141.0 C(5)CН/C(6),
8.04/155.3 C(5)CН/C(4) (the numeration shown on Scheme 1
was used).
2ꢀ[(4ꢀAnilinoꢀ3ꢀcyanoꢀ2ꢀoxoꢀ1,2ꢀdihydropyridinꢀ5ꢀyl)ꢀ
methylidene]aminoguanidine hydrochloride (3а•HCl) was preꢀ
pared from compound 1а according to the method B. M.p.
310—311 °С (DMF : PriOH, 1 : 1), the yield was 50%. ESI MS,
m/z: 296 [М + Н]+, 591 [2 М + Н]+. 1Н NMR (DMSOꢀd6), δ:
7.22—7.30 (m, 3 Н, Ph); 7.35—7.40 (m, 2 Н, Ph); 7.7 (br.s, 4 Н,
NНC(=NH)NH2); 8.07 (s, 1 Н, Н(6)); 8.11 (d, 1 Н, C(5)CН,
Jm = 2.9 Hz); 9.8 (br.s, 1 Н, С(4)NН); 11.47 (br.s, 1 Н, N(1)Н).
Found (%): С, 50.77; Н, 4.54; N, 29.50. С14H13N7OCl. Calcuꢀ
lated (%): C, 50.68; Н, 4.25; N, 29.55.
C. Concentrated HCl (0.05 mL) in water (0.05 mL) was
added to a solution of compound 5а (0.05 g, 0.12 mmol) in 96%
ethanol (1 mL) with stirring under reflux. The reaction mixture
obtained was refluxed with stirring for 2 h (the precipitate was
not dissolved), and was cooled to 20 °С. The precipitate was
filtered off and washed with 96% ethanol. Compound 3а was
obtained as hydrochloride in the yield of 0.013 g (26%).
2ꢀ{[4ꢀ(4ꢀChloroanilino)ꢀ3ꢀcyanoꢀ2ꢀoxoꢀ1,2ꢀdihydropyridinꢀ
5ꢀyl)methylidene}aminoguanidine (3b) was prepared from comꢀ
pound 1b according to the method А. The reaction time was
32 h, m.p. 335 °С (PriOH), the yield was 67%. ESI MS, m/z: 330
[М + Н]+, 659 [2 М + Н]+. 1Н NMR (DMSOꢀd6), δ: 5.49, 5.72
(both br.s, 2 Н each, NНC(=NH) NH2); 7.37 (d, 2 Н, Ar,
Jо = 8.6 Hz); 7.42 (d, 2 Н, Ar, Jо = 8.6 Hz); 7.77, 8.04 (both s,
1 Н each, Н(6), C(5)CН); 11.47 (br.s, 1 Н, С(4)NН). Found (%):
N, 30.05. С14H12N7OCl. Calculated (%): N, 29.73.
Synthesis and physicochemical characteristics of compounds
4ꢀAnilinoꢀ5ꢀformylꢀ2ꢀoxoꢀ1,2ꢀdihydropyridineꢀ3ꢀcarbonitrile
1
(1а)10. Н NMR (DMSOꢀd6), δ: 7.20—7.50 (m, 2 Н, Ph); 8.42
(s, 1 Н, Н(6)); 9.60 (s, 1 Н, СНО); 10.60 (br.s, 1 Н, N(1)Н);
12.46 (s, 1 Н, С(4)NН). 13С NMR (DMSOꢀd6), δ: 78.0 (С(3));
107.4 (С(5)); 113.9(С≡N); 124.7, 125.9, 128.6, 137.6 (Ph); 152.6
(С(6)); 155.6 (С(4)); 161.4 (С(2)); 190.7 (СНО).
Synthesis of compounds 3а,b and 4а,b (general procedure).
Method А. A mixture of compound 1a,b or 2a,b (2 mmol) and
aminoguanidinium hydrogencarbonate (4 mmol) in pyridine
(10 mL) was refluxed for 24 h (for 1a), the necessary reaction
time was determined using TLC. After cooling the reaction mixꢀ
ture to 20 °С, the precipitate was filtered off, washed with isoꢀ
propyl alcohol and petroleum ether.
2ꢀ{[4ꢀ(4ꢀChloroanilino)ꢀ3ꢀcyanoꢀ2ꢀoxoꢀ1,2ꢀdihydropyridinꢀ
5ꢀyl)methylidene}aminoguanidine hydrochloride (3b•HCl) was
prepared from compound 1b according to the method B. M.p.
328 °С (DMF), the yield was 90%. ESI MS, m/z: 330 [М + Н]+,
1
659 [2 М + Н]+. Н NMR (DMSOꢀd6), δ: 7.29, 7.37 (both d,
2 Н each, ClС6Н4); 7.69 (br.s, 4 Н, NНC(=NH)NH2); 8.10,
8.12 (both s, 1 Н each, Н(6), C(5)CН); 9.78 (br.s, 1 Н, С(4)NН);
12.00 (br.s, 1 Н, N(1)Н). Found (%): С, 45.77; Н, 3.13; N, 26.78.
С14H13N7OCl2. Calculated (%): C, 45.92; Н, 3.58; N, 26.77.
2ꢀ[4ꢀAnilinoꢀ(2ꢀoxoꢀ1,2ꢀdihydropyridinꢀ3ꢀyl)methylidene]ꢀ
aminoguanidine (4а) was prepared from compound 2а accordꢀ
ingly to the method А. The reaction time was 5 h. M.p. 280—282 °С
B. Concentrated HCl (0.2 mL) in water (0.2 mL) was added
to a mixture of compound 1a,b or 2a,b (0.2 g, 0.84 mmol) and
aminoguanidinium hydrogencarbonate (0.2 g, 1.50 mmol) in 96%

Reactions of aminoguanidine and guanidine
Russ.Chem.Bull., Int.Ed., Vol. 59, No. 12, December, 2010
2255
(DMF), the yield was 65%. IR, ν/сm–1: 3325, 3169, 3147 (NН,
NН2), 1640 (СО). ESI MS, m/z: 271 [М + Н]+, 541 [2 М + Н]+.
1Н NMR (DMSOꢀd6), δ: 5.31, 5.49 (both br.s, 2 Н each,
NНC(=NH)NH2); 6.08 (d, 1 Н, Н(5), Jо = 7.5 Hz); 7.07 (d, 1 Н,
Н(6), Jо = 7.5 Hz); 7.14 (t, 1 Н, Ph, Jо = 8.1 Hz); 7.28 (d, 2 Н,
Ph, Jо = 8.1 Hz); 7.38 (t, 2 Н, Ph, Jо = 8.1 Hz); 8.47 (s, 1 Н,
С(3)СН); 10.89 (br.s, 1 Н, N(1)Н); 11.46 (br.s, 1 Н, С(4)NН).
2ꢀ[4ꢀAnilinoꢀ(2ꢀoxoꢀ1,2ꢀdihydropyridinꢀ3ꢀyl)methylidene]ꢀ
aminoguanidine hydrochloride (4а•HCl) was prepared from comꢀ
pound 2а according to the method B. The reaction time was 1 h.
M.p. 306—308 °С (DMF), the yield was 62%. ESI MS, m/z: 271
[М + Н]+, 541 [2 М + Н]+. 1Н NMR (DMSOꢀd6), δ: 5.82 (d, 1 Н,
Н(5), Jо = 7.5 Hz); 7.11 (d, 1 Н, Н(6), Jо = 7.5 Hz); 7.30,
7.41 (both m, 5 Н, Ph); 7.70 (br.s, 3 Н, C(=NH)NH2); 8.63
(s, 1 Н, С(3)СН); 9.77, 11.67 (both br.s, 1 Н each, С(4)NН,
NН2C(=NH)NН—N=); 11.06 (br.s, 1 Н, N(1)Н). Found (%):
С, 51.12; Н, 5.14; N, 27.53. С13H15N6OCl. Calculated (%):
C, 50.90; Н, 4.93; N, 27.40.
was kept for 10 days at 20 °C. Powder Xꢀray diffraction analysis
of the precipitate that formed was carried out.
Complex of 4ꢀ(4ꢀchloroanilino)ꢀ5ꢀformylꢀ2ꢀoxoꢀ1,2ꢀdihydroꢀ
pyridineꢀ3ꢀcarbonitrile with aminoguanidine (5b) was prepared
analogously from compound 1b according to the variant А. M.p.
310—312 °С. The yield was 69%. IR, ν/сm–1: 3435, 3335, 3285,
3263 (NН, NН2); 2198 (СN); 1672 (СНО); 1630 (СО). 1Н NMR
(DMSOꢀd6), δ: 4.63 (br.s, 2 Н, NH2); 7.17, 7,36 (both d, 2 Н each,
ClС6Н4, Jо = 8.6 Hz); 7.24 (br.s, 4 Н, NНC(=NH)NН2); 8.26
(s, 1 Н, Н(6)); 9.33 (s, 1 Н, СНО); 10.41 (br.s, 1 Н, С(4)NН).
Complex of 4ꢀanilinoꢀ5ꢀformylꢀ2ꢀoxoꢀ1,2ꢀdihydropyridineꢀ3ꢀ
carbonitrile with guanidine (6). А. A mixture of compound 1а
(0.1 g, 0.48 mmol) and diguanidinium carbonate (0.08 g, 0.43 mmol)
in pyridine (2 mL) was refluxed until formation of white volumiꢀ
nous precipitate (the reaction time was 30 min). After cooling
the reaction mass to 20 °С, the precipitate was filtered off and
washed carefully with isopropyl alcohol. Compound 6 with adꢀ
mixture of diguanidinium carbonate was obtained in a yield of
0.15 g. This precipitate was refluxed in acetonitrile (130 mL),
filtered off and compound 6 was obtained in a yield 0.1 g (45%).
M.p. 235—236 °С (acetonitrile). IR, ν/сm–1: 3425, 3311, 3232
(NН, NН2); 2204 (СN), 1664, 1657 (СО). 1Н NMR (DMSOꢀd6),
δ: 7.13—7.18 (m, 2 Н, Ph); 7.32 (t, 3 Н, Ph, Jо = 7.7 Hz);
7.30 (br.s, 5 Н, NН2C(=NH)NН2); 8.10 (s, 1 Н, Н(6)); 9.34
(s, 1 Н, СНО); 10.40 (br.s, 1 Н, С(4)NН). 1Н NMR
(DMSOꢀd6+CD3OD), δ: 7.15 (d, 2 Н, Ph, Jо = 7.7 Hz); 7.30
(t, 3 Н, Ph, Jо = 7.7 Hz); 8.06 (s, 1 Н, Н(6)); 9.32 (s, 1 Н, СНО).
B. A mixture of compound 1а (0.10 g, 0.42 mmol) and
diguanidinium carbonate (0.05 g, 0.25 mmol) in pyridine (2 mL)
was refluxed for 0.5 h. After cooling the reaction mass to 20 °С,
the precipitate was filtered off and washed carefully with isoproꢀ
pyl alcohol. Compound 6 was obtained in a yield of 0.11 g (58%).
The sample for powder Xꢀray diffraction analysis was obtained
in the following way: compound 6 (0.09 g) in acetonitrile
(130 mL) was refluxed and filtered twice, the filtrate was kept for
10 days at 20 °C. The crystals that formed were filtered off (the
yield was 0.012 g) and powder Xꢀray diffraction analysis of the
precipitate was carried out.
2ꢀ{[4ꢀ(4ꢀChloroanilino)ꢀ2ꢀoxoꢀ1,2ꢀdihydropyridinꢀ3ꢀyl)ꢀ
methylidene}aminoguanidine (4b) was prepared from compound
2b according to the method А. The reaction time was 15 h. M.p.
320 °С (PriOH), the yield was 41%. IR, ν/сm–1: 3325, 3169,
3147 (NН, NН2), 1640 (СО). ESI MS, m/z: 305 [М + Н]+, 609
1
[2 М + Н]+. Н NMR (DMSOꢀd6), δ: 5.34, 5.50 (both br.s,
2 Н each, NНC(=NH)NH2); 6.06 (d, 1 Н, Н(5), Jо = 7.5 Hz);
7.06 (d, 1 Н, Н(6), Jо = 7.5 Hz); 7.29 (d, 2 Н, Ar, Jо = 8.8 Hz);
7.33 (d, 2 Н, Ar, Jо = 8.8 Hz); 8.45 (s, 1 Н, С(3)СН); 10.89 (br.s,
1 Н, N(1)Н); 11.43 (br.s, 1 Н, С(4)NН).
Complex of 4ꢀanilinoꢀ5ꢀformylꢀ2ꢀoxoꢀ1,2ꢀdihydropyridineꢀ3ꢀ
carbonitrile with aminoguanidine (5а). А. A mixture of compound
1а (0.1 g, 0.42 mmol) and aminoguanidinium hydrogencarbonꢀ
ate (0.11 g, 0.84 mmol) in pyridine (2 mL) was refluxed until
formation of white voluminous precipitate (the reaction time
was 30 min). After cooling the reaction mass to 20 °С, the preꢀ
cipitate was filtered off and washed carefully with isopropyl alꢀ
cohol and water. Compound 5а was obtained in a yield of 0.11 g.
M.p. 314—315 °С (acetonitrile), the yield after recrystallization
was 45%. IR, ν/сm–1: 3452, 3288, 3142 (NН, NН2); 2206 (СN);
1681 (СНО); 1637 (СО). 1Н NMR (DMSOꢀd6), δ: 4.67 (br.s, 2 Н,
NH2); 7.13—7.18 (m, 2 Н, Ph); 7.32 (t, 3 Н, Ph, Jо = 7.7 Hz);
7.30 (br.s, 4 Н, NНC(=NH)NН2); 8.13 (s, 1 Н, Н(6)); 9.34
(s, 1 Н, СНО); 10.41 (br.s, 1 Н, N(1)Н). 13С NMR (DMSOꢀd6),
δ: 79.5 (С(3)); 106.6 (С(5)); 117.5 (С≡N); 124.3, 125.1, 128.5,
138.2 (Ph); 155.2 (С(4)); 162.9 (С(6)), 172.9 (С(2)); 188.6
(CНО). The assignment of the signals in the 13С NMR spectrum
of compound 5а was made based on the HMBC technique. Charꢀ
acteristic correlation peaks in the HMBC spectrum of compound
5а are as follows: 8.13/106.6 Н(6)/C(5), 8.13/155.2 Н(6)/C(4),
8.13/172.9 Н(6)/C(2), 8.13/188.6 Н(6)/СНО, 9.35/106.6 CНО/
C(5), 9.35/155.2 CНО/C(4) 9.35/162.9 CНО/C(6) (the numerꢀ
ation shown in Scheme 1 was used). Found (%): С, 53.34;
Н, 4.76; N, 31.28. С14H15N7O2. Calculated (%): C, 53.67;
Н, 4.83; N, 31.29.
4ꢀAnilinoꢀ2ꢀoxoꢀ1,2ꢀdihydropyridine (7). A. A mixture of
compound 2а (0.15 g, 0.7 mmol) and diguanidinium carbonate
(0.13 g, 0.72 mmol) in pyridine (3 mL) was refluxed for 22 h.
After cooling the reaction mass to 20 °С, the precipitate was
filtered off and washed carefully with isopropyl alcohol. The
crude product 7 was obtained in a yield of 0.08 g. The mother
liquor was concentrated in vacuo, the crystalline residue was
triturated with isopropyl alcohol, filtered off, and washed with
isopropyl alcohol. The crude product 7 was additionally obtained
in a yield of 0.11 g. The yield of recrystallized product was 33%.
M.p. 215—218 °С (acetonitrile). ESI MS, m/z: 187 [М + Н]+,
1
209 [М + Na]+, 395 [2 М + Na]+, 581[3 М +Na]+. Н NMR
(DMSOꢀd6), δ: 5.65 (d, 1 Н, Н(3), 4J = 2.0 Hz); 5.86 (d.d, 1 Н,
Н(5), 3J = 7.2 Hz, Jm = 2.0 Hz); 7.01 (t, 1 Н, Ph, Jо = 7.7 Hz);
7.03 (d, 1 Н, Н(6), 3J = 7.2 Hz); 7.15 (d, 2 Н, Ph, Jо = 7.7 Hz);
7.31 (t, 2 Н, Ph, Jо = 7.7 Hz); 8.47 (br.s, 1 Н, С(4)NН); 10.56
(br.s, 1 Н, N(1)Н). Found (%): С, 70.63; Н, 5.68; N, 15.09.
С11H10N2O. Calculated (%): C, 70.97; Н, 5.38; N, 15.05.
B. KOH (1 g, 18 mmol) and ethylene glycol (10 mL) were
added to pyridone 8 (1 g, 4.7 mmol), and the mixture obtained
was refluxed for 6 h. Then it was cooled, diluted with water
(40 mL) and kept at 20 °С, the precipitate was filtered off and
washed carefully with water. The crude product 7 was obtained
B. A mixture of compound 1а (0.1 g, 0.42 mmol) and aminoꢀ
guanidinium hydrogencarbonate (0.057 g, 0.42 mmol) in pyriꢀ
dine (2 mL) was refluxed for 2 h. The precipitate was filtered off
after cooling the reaction mass to 20 °С and washed carefully
with isopropyl alcohol. Compound 5а was obtained in a yield of
0.076 g (58%). The sample for powder Xꢀray diffraction analysis
was obtained in the following way: compound 5а (0.05 g) in
acetonitrile (100 mL) was refluxed and filtered twice, the filtrate

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Medvedeva et al.
in a yield of 0.75 g. The yield of recrystallization product was
41%. M.p. 228—230 °С (ethanol).
precipitate with admixture of inorganic impurities decomposed
upon attempted recrystallization.
1,3ꢀBis[(dimethylamino)methylidene]ꢀ2ꢀ[(4ꢀanilinoꢀ3ꢀcyꢀ
anoꢀ2ꢀoxoꢀ1,2ꢀdihydropyridinꢀ5ꢀyl)methylidene]aminoguanidine
(9а). А. A solution of compound 3а as a free base (0.02 g,
0.68 mmol) and dimethylformamide dimethylacetal (0.35 g,
3.0 mmol) in isopropyl alcohol (2 mL) was kept at 20 °С for
3 days. The solvent and excess of the acetal were evaporated
in vacuo, the residue was triturated with petroleum ether, the
precipitate that formed was filtered off and washed with petroꢀ
leum ether. Compound 9а was obtained in a yield of 0.02 g
(74%). M.p. 238—240 °С. ESI MS, m/z: 406 [М + Н]+, 428
1,3ꢀBis[(dimethylamino)methylidene]ꢀ2ꢀ[(4ꢀanilinoꢀ3ꢀcyꢀ
anoꢀ1ꢀmethylꢀ2ꢀoxoꢀ1,2ꢀdihydropyridinꢀ5ꢀyl)methylidene]ꢀ
aminoguanidine (9b). А. Me2NCH(OMe)2 (0.45 g, 3.8 mmol)
was added to a suspension of compound 3а as a free base (0.2 g,
0.68 mmol) in dry toluene (15 mL). The reaction mixture was
refluxed with stirring for 6 h, cooled to 20 °С, the precipitate was
filtered off, triturated with isopropyl alcohol, filtered off, and
washed with petroleum ether. Analytically pure compound 9b
was obtained in a yield of 0.09 g (34%). M.p. 221—222 °С. ESI
MS, m/z: 420 [М + Н]+, 442 [М + Na]+, 839 [2 М + Н]+, 861
1
1
[М + Na]+, 811 [2 М+ Н]+. Н NMR (DMSOꢀd6), δ: 2.34,
[2 М + Na]+. Н NMR (DMSOꢀd6), δ: 2.37, 2.93, 3.00, 3.11,
2.87, 2.94, 3.04 (all s, 3 Н each, 4 СН3); 7.24—7.30, 7.35—7.39
(both m, 5 Н, Ph); 7.75, 8.27 (both s, 1 Н each, Н(6); С(5)СН);
8.20, 8,21 (both s, 1 Н each, 2 ((СН3)2NCН(=N)); 11.50 (br.s,
1 Н, N(1)Н); 12.94 (br.s, 1 Н, С(4)NН).
B. It was prepared from hydrochloride of compound 3а acꢀ
cording to the method А. A suspension was kept at 20 °С for
13 days. The precipitate was filtered off and washed with isoproꢀ
pyl alcohol. Compound 9а was obtained in a yield of 0.014 g
(50%), m.p. 238—240 °С.
C. It was prepared from hydrochloride of compound 3а acꢀ
cording to the method А. A suspension was kept at 20 °С in
toluene for 48 h. The precipitate was filtered off and washed with
toluene and petroleum ether. The crude product 9а was obꢀ
tained in a yield of 0.02 g (79%), m.p. 190 °С (decomp.). The
3.43 (all s, 3 Н each, 5 СН3); 7.24—7.30, 7.33—7,39 (both m,
5 Н, Ph); 7.99, 8.16 (both s, 1 Н each, Н(6), С(5)СН); 8.22,
8,24 (both s, 1 Н each, 2 ((СН3)2NCН(=N)); 12.89 (br.s, 1 Н,
С(4)NН). Found (%): С, 59.64; Н, 5.80; N, 29.44. С21H25N9O.
Calculated (%): C, 60.13; Н, 6.01; N, 30.05.
B. Triethylamine (0.067 g, 0.7 mmol) was added to a suspenꢀ
sion of hydrochloride of compound 3а (0.02 g, 0.07 mmol) in dry
toluene (2 mL) and the reaction mixture was stirred for 10 min.
Then Me2NCH(OMe)2 (0.22 g, 1.9 mmol) was added dropwise
and the reaction mass was refluxed with stirring for 2 h. The
solvent and excess of the acetal were evaporated in vacuo, the
residue was triturated with isopropyl alcohol, the precipitate that
formed was filtered off and washed with isopropyl alcohol; anaꢀ
lytically pure compound 9b was obtained in a yield of 0.01 g
Table 7. Crystallographic characteristics of compounds 3a, 5a, and 6
Parameter
3а
5а
6
Molecular formula
Crystal system
Space group
C14H13N7O
Моноклинная
P21/c
C13H9N3O2•CH6N4
Моноклинная
P21/c
C13H9N3O2•CH5N3
Моноклинная
P21/c
a/Å
b/Å
c/Å
α/deg
β/deg
γ/deg
V/Å3
13.9356(12))
4.8199(4)
21.6685(19)
90
100.96(3)
90
6.2161(11)
8.2131(13)
30.309(4)
90
104.72(3)
90
5.4448(5)
17.4721(19)
15.3241(18)
90
103.14(3)
90
1428.9(3)
27
1496.4(4)
21
1419.5(3)
30
M20
*
F30
*
33 (0.010, 47)
34 (0.011, 51)
41 (0.009, 53)
Z
4
4
4
Dx/g cm–3
1.373
1.391
1.396
Radiation
(waveꢀlength/Å)
Powder diagram:
CuKa1
(1.5406)
4.00—80.00
CuKa1
(1.5406)
4.00—75.00
CuKa1
(1.5406)
5.00—80.00
2θmin – 2θ
max
(step of measurement/deg)
0.01
0.01
0.01
Rp*
Rwp
0.0236
0.0308
0.0130
5.176
0.0142
0.0177
0.0142
1.604
0.0259
0.0334
0.0185
4.252
*
*
Rexp
2
χ *
* Indicators of indication quality M20 (see Ref. 26) and F30 (see Ref. 27) were determined previously, Rp, Rwp
Rexp and χ — see Ref. 28.
,
2

Reactions of aminoguanidine and guanidine
Russ.Chem.Bull., Int.Ed., Vol. 59, No. 12, December, 2010
2257
(34%). M.p. 221—222 °С. ES MS, m/z: 420 [М + Н]+, 442 [М +
Na]+, 839 [2 М + Н]+, 861 [2 М + Na]+.
cule and one for the molecule of 2ꢀaminoguanidine. Hydrogen
atoms were placed at the calculated positions and were not
refined. The principal crystallographic characteristics and exꢀ
perimental parameters of compounds 3а, 5а, and 6 are given in
1,3ꢀBis[(dimethylamino)methylidene]ꢀ2ꢀ{[4ꢀ(4ꢀchloroaniliꢀ
no)ꢀ3ꢀcyanoꢀ1ꢀmethylꢀ2ꢀoxoꢀ1,2ꢀdihydropyridinꢀ5ꢀyl]methylꢀ
iden}aminoguanidine (9c) was prepared analogously from comꢀ
pound 9a according to the method А. The yield was 83%. M.p.
260 °С (decomp.). ESI MS, m/z: 454 [М + Н]+, 476 [М + Na]+,
907 [2 М + Н]+. 1Н NMR (DMSOꢀd6), δ: 2.42, 2.93, 2.97, 3.07,
3.41 (all s, 3 Н each, 5 СН3); 7.27 (d, 2 Н, Ar, Jо = 8.5 Hz); 7.41
(d, 2 Н, Ar, Jо = 8.5 Hz); 8.05, 8.43 (both s, 1 Н each, Н(6),
С(5)СН); 8.21, 8,25 (both s, 1 Н each, 2 ((СН3)2NCН(=N));
12.91 (br.s, 1 Н, С(4)NН). Found (%): С, 55.70; Н, 5.55;
N, 27.67. С21H24N9OCl. Calculated (%): C, 55.57; Н, 5.33;
N, 27.77.
a
I
45000
×4
1,3ꢀBis[((dimethylamino)methylidene]ꢀ2ꢀ[(4ꢀanilinoꢀ2ꢀoxoꢀ
1,2ꢀdihydropyridinꢀ3ꢀyl)methylidene]aminoguanidine (10). А.
Me2NCH(OMe)2 (0.26 g, 2.2 mmol) was added to a suspension
of compound 4а as a free base (0.06 g, 0.22 mmol) in PriOH
(3 mL). The reaction mass was kept at 20 °С for 5 days. The
precipitate was filtered off and washed with PriOH. The crude
product 12 was obtained in a yield of 0.05 g (61%). M.p.
231—234 °С. ESI MS, m/z: 381 [М + Н]+, 761 [2 М + Н]+.
1Н NMR (DMSOꢀd6), δ: 2.86, 2.94 (both s, 3 Н each, 2 СН3);
3.05, (s, 6 Н, 2 СН3); 5.87 (d, 1 Н, Н(5), Jо = 7.5 Hz); 7.19
(d, 1 Н, Н(6), Jо = 7.5 Hz); 7.21, 7.41 (both m, 5 Н, Ph); 8.21
(s, 2 Н, (СН3)2NCН(=N); 8.70 (s, 1 Н, С(3)СН); 10.80 (br.s,
1 Н, N(1)Н); 12.80 (br.s, 1 Н, С(4)NН).
1
0
2
10 20
30
40 50 60
70 80 2θ/deg
b
I
45000
B. It was prepared from compound 4а as a free base accordꢀ
ing to the method А. The solution in dry toluene was refluxed for
6 h. The precipitate that formed after keeping the reaction mixꢀ
ture at 20 °С for 48 h was filtered off and washed with toluene.
The crude product 10 was obtained in a yield of 0.024 g (29%),
m.p. 240—241 °С. The mass spectrum of this sample points to
the presence of traces of Nꢀmethylated (at the pyridine nitrogen
atom) product. ESI MS, m/z: 395 [М + Н]+.
×4
1
2
0
C. A mixture of hydrochloride of compound 4а (0.022 g,
0.07 mmol) and Me2NCH(OMe)2 (0.35 g, 3 mmol) in dry toluꢀ
ene (1 mL) was kept at 20 °С for 22 h. The precipitate was
filtered off, washed with toluene and petroleum ether. The crude
product 10а was obtained in a yield of 0.026 g (95%), m.p.
212—214 °С. The precipitate was contaminated with inorganic
impurities and decomposed upon attempted recrystallization.
XꢀRay diffraction analysis of compounds 3а, 5а, and 6. The
structures of compounds 3а, 5а, and 6 were established by powꢀ
der Xꢀray diffraction.20 Powder diagram was measured in the
Guinier camera «Huber G670» containing bent germanium
monochromator. The positions of first 30 peaks were refined on
the powder diagrams. Using these positions, indicating in the
triclinic cell was carried out with the TREOR90 program.21 The
crystal structures of compounds 3а, 5а, and 6 were solved by the
method of simulated annealing.22 Threeꢀdimensional molecule
model obtained as a result of the optimization by the density
functional method using the PRIRODA program23 was used.
Then the solution obtained was refined by the Rietveld method
using the MRIA program,24 peak profiles were described by the
modified Voight function.25 In the refinement, restriction to the
permissible deflections of the interatomic distances in the moleꢀ
cule and to the planarity of the rings were applied. Parameters of
thermal fluctuations (Uiso) of nonhydrogen atoms in 3а and 6
were refined in an isotropic approximation. For only two generꢀ
al parameters Uiso were refined, viz., one for the main moleꢀ
10
20 30 40
50 60 70 80 2θ/deg
c
I
15000
×2
1
0
2
5
15
25 35
45 55
65
75 2θ/deg
Fig. 3. The result of elucidation of crystal structures of comꢀ
pounds 3a (a), 6 (b), and 5a (c) by the Rietveld method: experiꢀ
mental curve (1), the difference between the experimental and
calculated curves as a result of refinement (2); highꢀangle area
(2θ > 35°) is presented on a large scale. The calculated positions
of reflexes are indicated as vertical segments.

2258
Russ.Chem.Bull., Int.Ed., Vol. 59, No. 12, December, 2010
Medvedeva et al.
Table 7. The experimental powder diagram and the difference
curve as a result of refinement using the Rietveld method are
shown in Fig. 3.
All structural data were deposited with the Cambridge Crysꢀ
tallographic Database (CCDC 785159—785161)*.
menko, V. G. Granik, Izv. Akad. Nauk, Ser. Khim., 2009,
2265 [Russ. Chem. Bull., Int. Ed., 2009, 58, 2336].
12. F. H. Allen, Acta Crystallogr. Sect. B: Struct. Sci., 2002,
58, 380.
13. M. Feelish, P. Kotsonis, J. Siebe, B. Clement, H. Schmidt,
Mol. Pharmacol., 1999, 56, 243.
This work was financially supported by the Russian
Foundation for Basic Research (Project No. 10ꢀ03ꢀ00061ꢀa).
14. M. O. Ribeiro, E. Antunes, G. de Nucci, S. M. Lovisolo,
R. Zatz, Hypertension, 1992, 3, 298.
15. N. Rakieten, M. L. Rakieten, M. V. Nadkarni, Cancer
Chemother, 1963, 29, 91.
16. R. Koster, M. Anderson, E. J. de Beer, Fed. Proc., 1959,
18, 412.
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* Copies can be obtained, free of charge, on application to the
Cambridge Crystallographic Data Centre (CCDC), 12 Union
Road, Cambrige CB2 1EZ, UK. Fax: +44 (0) 1223 336033.
Eꢀmail: deposit@ccdc.cam.ac.uk.
Received August 12, 2010