Synthesis of 4-amino-2,6-diaryl-5-cyanopyrimidines as antimicrobial agents

Article abstract of DOI:10.1080/00397911.2016.1151051

This article describes an efficient and facile synthesis of 11 2,4,6-trisubstituted 5-cyanopyrimidines starting from meta- and para-substituted 2-cyanocinnamonitriles and arylamidines. The synthesized heterocycles, 3a–k, were characterized by infrared (IR

Full text of DOI:10.1080/00397911.2016.1151051

Synthetic Communications
An International Journal for Rapid Communication of Synthetic Organic
Chemistry
ISSN: 0039-7911 (Print) 1532-2432 (Online) Journal homepage: http://www.tandfonline.com/loi/lsyc20
Synthesis of 4-amino-2,6-diaryl-5-
cyanopyrimidines as Antimicrobial Agents
Zenaide Severina do Monte, Maria Renata Leite Monteiro, Camila Beatriz
Atanásio Borba, Norma Buarque de Gusmão, Emerson Peter da Silva Falcão,
Ricardo Oliveira Silva, Rajendra M. Srivastava & Sebastião José de Melo
To cite this article: Zenaide Severina do Monte, Maria Renata Leite Monteiro, Camila Beatriz
Atanásio Borba, Norma Buarque de Gusmão, Emerson Peter da Silva Falcão, Ricardo
Oliveira Silva, Rajendra M. Srivastava & Sebastião José de Melo (2016): Synthesis of 4-
amino-2,6-diaryl-5-cyanopyrimidines as Antimicrobial Agents, Synthetic Communications, DOI:
10.1080/00397911.2016.1151051
To link to this article: http://dx.doi.org/10.1080/00397911.2016.1151051
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Date: 25 February 2016, At: 14:18

Synthesis of 4-amino-2,6-diaryl-5-cyanopyrimidines as antimicrobial agents
1
,2
2
Zenaide Severina do Monte , Maria Renata Leite Monteiro , Camila Beatriz Atanásio
2
2
3
Borba , Norma Buarque de Gusmão , Emerson Peter da Silva Falcão , Ricardo Oliveira
4
4,+
1,2,
Silva , Rajendra M. Srivastava , Sebastião José de Melo
1
Department of Pharmaceutical Sciences, Federal University of Pernambuco (UFPE),
2
Recife, PE, Brazil, Department of Antibiotics, Federal University of Pernambuco
3
(
5
UFPE), Recife, PE, Brazil, Department of Nutrition, (UFPE), Núcleo de Nutrição CAV
5608-680 - Vitoria de Santo Antão, PE, Brazil, Department of Fundamental Chemistry,
4
(UFPE), Recife 50740-540, PE, Brazil
Corresponding author: Sebastião José de Melo, Department of Antibiotics, Federal
University of Pernambuco, Recife 50740-560, PE, Brazil. E-mail:
melosebastiao@yahoo.com.br
+
Visiting Research Scientist, Department of Antibiotics, UFPE, Recife, PE, from
December 1, 2014 through February 28, 2015. This visit was financed by Fundação de
Amparo à Ciência e Tecnologia do Estado de Pernambuco(FACEPE), PE, Brazil.
Abstract
This paper describes an efficient and facile synthesis of eleven 2,4,6-trisubstituted 5-
cyanopyrimidines starting from meta and para substituted 2-cyanocinnamonitriles and
1
arylamidines. The synthesized heterocycles, 3a-k, were characterized by IR, H-NMR,
1
3
C-NMR and mass spectral data. The probable mechanism of formation of the title
compounds employing 2-cyanocinnamonitriles and arylamidines in the presence of a base
was clarified.
A preliminary screening of the antibacterial tests clearly showed that four out of eleven
pyrimidines, viz., 3a, 3e, 3f and 3k were effective against bacteria, Staphyloccus aureus,
Bacillus subtillis and Pseudomonas aeruginosa. Further, the Minimum Inhibitory
Concentration (MIC) against the bacteria has been determined.
1

KEYWORDS: arylpyrimidines, 2-cyanocinnamonitriles, 4-amino-2,6-diaryl-5-
cyanopyrimidine, reaction mechanism, antimicrobial agents
1. INTRODUCTION
Extensive investigations of pyrimidines indicate that they are substructures of crucial
building blocks of nucleotides with uracil and thymine, being the constituents of
ribonucleic acid (RNA) and deoxyribonucleic acid (DNA), respectively. Thymine is one
of the nucleobases in the nucleic acid of DNA. In RNA, the thymine is replaced by the
nucleobase uracil, a demethylated thymine. Pyrimidines containing compounds occur in
[
1]
nature and have biological properties . These include nucleosides as well as alkaloids
and antibiotics. Substituted pyrimidines prepared synthetically have been found to
[
2]
possess a variety of biological activities. For example, some are antiviral agents , others
[3-5]
possess anti-parasitic, antimicrobial, anticancer and antibiotic activities . In 2008, we
2

reported the synthesis and antiinflammatory activity of 4-amino-2,6-diaryl-5-
cyanopyrimidines, where three compounds were found to posses antiinflammatory
[6]
activity which are twice more effective than aspirin in reducing inflammation . A
recent review covers the significance and importance of pyrimidines in the Microbial
[
7]
World .
Considering observations in the recent literature, we prepared eleven 4-amino-2,4-diaryl-
5
-cyanopyrimidines with a view to verify their structures and evaluate their antibacterial
properties. The present paper describes the synthesis, mechanism of formation and
biological activity of title compounds., These compounds produced reasonable activity
against bacteria.
2
. RESULTS AND DISCUSSION
The starting materials, arylamines, 1a-k, and 2-cyanocimmonitriles, 2a-k, were
[
6,8]
prepared according to the published procedure described earlier
f-g were individually dissolved in methanol and a catalytic quantity of piperidine was
added, followed by reflux for 6 hours. The work-up produced the desired products 3f and
g in 92% and 95% yields, respectively. Similarly, 1a-e/1h-k and 2a-e/2h-k were
. Compounds 1f-g and
2
3
allowed to react individually providing the desired compounds 3a-e and 3h-k in excellent
yields (89-98%) as shown in Scheme S1. We were able to synthesize eleven 4-amino-2,6-
diaryl-5-cyanopyrimidines 3a-k in their crystalline states.
3

Eight of the title compounds, i.e., 3c, e-k have not yet been reported in the literature.
[6,9]
Compounds 3a, 3b and 3d
have been described in the literature, but in smaller yields
by using a different base and a longer reaction time (8 hours). Substances 3a, 3b and 3d
have been obtained in excellent yields in the present work. In fact, 3a and 3b gave almost
quantitative yields (98%), while 3d in 91% yield, respectively. Here the reaction time
[
6]
was 6 hours. Peter et al. synthesized the compounds 3b and 3d with yields of 33% and
3
2%, respectively, using the reaction time of 8 hours. Compounds 3a and 3b were also
[
9]
obtained by Xavier et al. with yields of, 40% and 67%, respectively, employing
microwave radiation with reaction time of 40 minutes, in the presence of potassium
carbonate as a base.
1
All compounds were characterized by their spectroscopic data, such as, IR, H-NMR,
1
3
C-NMR and mass spectra. Considering that all compounds have three aromatic rings
1
13
and eight quaternary carbons, H and C NMR assignments were made using 2D
1
1
experiments: H- H COSY (Correlation Spectroscopy), Heteronuclear Multi-Bond
Correlation (HMBC) and Heteronuclear Single-Quantum Coherence (HSQC)
1
13
spectroscopy. Supporting information of this paper, presents the H and C NMR
1
13
spectra of compounds 3a, 3d, 3f-g and 3k. By looking at H and C NMR spectra, it
1
1
seemed quite difficult to attribute all signals. The H- H COSY spectrum of compound
d made it possible to identify two para-substituted phenyl systems. The methoxy group
3
was assigned easily at δ 3.81 ppm. From HSQC and HMBC spectra, it was possible to
assign all hydrogen and carbon nuclei in the p-methoxyphenyl group. A similar strategy
4

was used to attribute the signals of the p-nitro- and m-nitrophenyl group of all
compounds.
Here follows a description of the NMR assignments of 3f-g. Initially, we examined the
1
H NMR spectrum of compound 3f, which presented a singlet at δ 3.01 ppm, which is
assigned as H1’’’, and four signals at δ 6.77, 7.64, 7.97 and 8.24 ppm. We observed two
COSY correlations: δ 6.77 with δ 8.24 ppm; and δ 7.64 with δ 7.97. Therefore, there are
two AA’BB’ systems, with all showing a coupling constant equal to 9.30 Hz. Also one
band centered at δ 7.73 ppm was observed and assigned to the hydrogen nuclei of the
1
3
amino group. The C NMR spectrum presented 14 signals. The signal at δ 39.7 ppm
C
was easily attributed to methyl groups. To assign all the others, it was necessary to obtain
2
D spectra. In the HSQC spectrum, the following correlations to the AA’BB’ systems
were observed: δ 6.77 ppm – δ 111.1 ppm; δ 7.64 ppm – δ 128.5 ppm; δ 7.97 ppm –
H
C
H
C
H
δ 130.4 ppm; and δ 8.24 ppm – δ 130.1 ppm. But a complete assignment was only
C
H
C
possible using the HMBC experiment. In this spectrum, correlations were observed from
the signal attributed to methyl group (δ 3.01 ppm) with the following signals: δ 152.6
H
C
ppm, assigned to C4’ nucleus, and δ 39.7 ppm. Table S1 presents all HMBC correlations
C
observed and the attributions made.
The signals at δ 164.3, 116.8 and 82.4 ppm were not yet attributed. Also the C4, C5 and
C
C1’’’’ nuclei were not assigned. Considering the structure of compound 3f, we attributed
the signal at δ 82.4 to the C5 nucleus because its proximity to a sp hybridized carbon.
C
5

The C1’’’’ nuclei are shifted downfield from the C4 nucleus, making the signal at δC
64.30 ppm which can be attributed to the C4 nucleus.
1
1
Next, attribution to compound 3g was performed. In the H NMR spectrum, the singlet at
δ 2.38 ppm was attributed to the methyl group. There is one band, centered at δ 8.06 ppm,
1
1
assigned to hydrogen nuclei of amino group. The H- H COSY spectrum shows two spin
systems, which are constituted by signals centered at δ 8.75, 8.42 and 7.89 ppm, for the
first system; and δ 8.29 and 7.34 ppm, for the second system. The signal at δ 8.75 ppm (t,
2
1
.1 Hz, 1H) was attributed to the H2’’ nucleus, while the peak at δ 7.89 ppm (t, 8.1 Hz,
H) was assigned to H5’’ nucleus. The assignment made to H2’’ nucleus is explained by
as follows: first, the downfield shift appears due to its structure where the just-mentioned
proton has a nitro group in its ortho position and the pyrimidine ring is also linked at
ortho to this hydrogen (Linkage 1” → 6, see Figure S1). Since the aromatic proton H2”
has two electro-withdrawing groups – the pyrimidine ring on one side and the nitro group
on the other side, this aromatic hydrogen will have a strong deshielding effect. Further,
assuming the planar structure of the molecule, the lone pair of electron on N1 plays an
additional influence on H2” causing a further downfield shift.
Another observation is about the smaller peak area is assigned to H2” nucleus. Since
there is no proton in its vicinity, its relaxation process is quite slow. This explains the
lower integrated area as well for this proton.
6

The multiplet among δ 8.47 – 8.39 ppm (2H) was assigned to the H4’’ and H6’’ nuclei.
The two duplets at δ 8.29 ppm and δ 7.34 ppm were attributed to AA’BB’ system, but
1
3
they were not assigned. In the C spectrum, the signal at δ 21.2 ppm was attributed to the
methyl group, while the signal at δ 84.7 ppm was attributed to the C5 nucleus. In the
HMBC spectrum, the signal at δ 21.2 ppm is correlated to the duplet at δ 7.34 ppm. So,
the signal at δ 7.34 ppm was attributed to the H3’and H5’ nuclei, and the signal at δ 8.29
ppm was attributed to the H2’ and H6’ nuclei. Since the hydrogen nuclei of the AA’BB’
system were already attributed, it is possible assign the carbon nuclei in this system,
using HMBC and HSQC spectra. The signals at δ 129.3 and 128.5 ppm were attributed to
the C3’/C5’ and C2’/C6’ nuclei, respectively. The signal attributed to the H3’/H5’ nuclei
shows an HMBC correlation with the signals attributed to the methyl group, C3’/C5’ and
the C1’ nuclei. Therefore, the signal at δ 133.6 ppm was attributed to C1’ nucleus. The
peak assigned to the H2’/H6’ nuclei presents an HMBC correlation with the signals
attributed to C2, C2’/C6’ and C4’. Then, the signals at δ 164.3 and 141.9 ppm were
attributed to the C2 and C4’ nuclei, respectively.
The first spin system, observed in COSY spectrum, was attributed to the m-NO2
substituted ring. HMBC and HSQC correlations were used to attribute the carbon nuclei
to this system. The signals at δ 123.4 and 130.4 ppm were attributed to the C2’’ and C5’’
nuclei, respectively. To attribute the H4’’, H6’’, C4’’ and C6’’ nuclei, we use HMBC
correlations. H4’’ nuclei shows the HMBC correlation with the signals attributed to the
C6’’ and C2’’ nuclei, while the H6’’ nuclei presents an HMBC correlation with the C6,
C2’’ and C4’’ nuclei. Therefore, the signals at δ 8.47 – 8.39 ppm were attributed to H6’’
7

and H4’’, respectively. The signals at δ 125.5 and 135.1 ppm were attributed to C6’’ and
C4’’, respectively. The signal at δ 166.0 ppm was attributed to the C6 nucleus. The peak
attributed to H5’’ presents HMBC correlations with the C3’’ and C1’’ nuclei. On the
other hand, the C3’’ nucleus shows a weak HMBC correlation with the H4’’and H2’’
nuclei. Then, the signals at δ 147.8 and 138.1 ppm were attributed to the C3’’ and C1’’
nuclei, respectively. Finally, the signals at δ 164.4 and 116.1 ppm were attributed to the
CN group and the C4 nucleus, respectively.
2
.1 Mechanism Of Formation Of 4-Amino-2,6-Diaryl-5-Cyanopyrimidines
[
10]
In 2000, Dos Santos proposed the mechanism of formation of 4-amino-2,6-diaryl-5-
cyanopyrimidines briefly, but did not publish it . Since then, two more articles have
appeared in the literature dealing with the mechanism of formation of 4-oxo-5-
[
10,11]
cyanopyrimidines
. In this present publication, we are trying to give more details and
clarification concerning the mechanism of formation of the title pyrimidines. These are
given below:
Initially, we tried to see if the reaction could be completed without the use of a base. For
this, we used reagents 1a and 2a in methanol and refluxed the contents for 6 hours.
Examination of the contents by thin-layer chromatography clearly showed the absence of
any product formation. Only the starting reagents were observed. However, when a
catalytic quantity of piperidine was added to the reagents 1a and 2a in methanol followed
by reflux, product formation was clearly observed. Thus, it is clear that a base is
important to affect the reaction. The piperidine abstracts a proton from arylamidine to
8

'
produce a stable anion 1 , which then attacks the carbon-3 of 2-cyanocinnamonitrile 2, as
shown in scheme S2. The anion 6 abstracts a proton from 4 to give 7 where the imidine
nitrogen is properly set up to attack the carbon atom of one of the ––CtbndN group to
provide 8 which goes to 9. This intermediate then tautomerizes to 10, which is the 3,6-
dihydro-pyrimidine. This in the presence of air reacts with atmospheric oxygen to furnish
the final product 3a. Similarly, 3b-k are also formed.
We tried to isolate 10 by performing the reaction under nitrogen atmosphere, where the
thin-layer chromatogram had shown a slow-moving spot with R = 0.2 and another spot
f
at R = 0.4 (n-hexane-ethyl acetate, 7:3). This last spot was attributed to pyrimidine 3 and
f
not dihydropyrimidine 10. This slow moving spot is assumed to be due to 10. When the
contents were heated again without nitrogen, this slow moving spot disappeared as
indicated by TLC and the spot with R value 0.4 intensified. This is indicative of the
f
aromatization of 10 in the presence of air. It therefore appears that biradical oxygen is
responsible for the aromatization of 10 to 3.
2
.2. Antibacteria Activity
Compounds 3a-k were screened for antibacterial activity. (Mueller Hinton Broth
Difco) 3.2 to 0.00125 mg/mL was used as nutrient, with DMSO as solvent and a blank.
–
The compounds were tested for their activities against Gram positive bacteria
(Staphylococcus aureus and Bacillus subtillis) and Gram Negative bacteria
(Pseudomonas aeruginosa). The results revealed that most of the synthesized compounds
showed varying degrees of inhibition against the tested microorganisms. In general, the
9

inhibitory activity against the tested Gram Positive bacteria was higher than that of the
Gram Negative bacteria. In short, all pyrimidines 3a, e, f, g and k have bacterial growth
inhibitions, as shown in Table S2. Compounds 3a, e, f, and k displayed broad spectrum
antibacterial property. These were the best of the series, and presented activity against all
three organisms, at lower concentrations, varying between 1.6 to 0.1 mg /mL. Compound
3
c was only moderately active against Gram Negative bacteria (Pseudomonas
aeruginosa) at a concentration of 3.2 mg /mL.
3
. EXPERIMENTAL
3
.1. General Experimental Procedures
All reagents were obtained from commercial sources and used without further
purification. All melting points were recorded on a BUCHI B-540 apparatus and are not
corrected. The IR spectra were recorded on a Perkin Elmer Spectrum400, using the KBr
wafer technique. The spectra of compound 3b-d and 3f-k were acquired using VARIAN
1
13
VNMRS400 spectrometer operating at 400 MHz and 100 MHz for H and C nuclei,
respectively. The spectra of compounds 3a, 3e and 3g were performed using VARIAN
1
13
Unity Plus 300 spectrometer operating at 300 MHz and 75 MHz for H and C nuclei,
respectively. HSQC and HMBC experiments were optimized to observe coupling
1
3
.
constants equal to 140 Hz ( J ) and 8 Hz ( J ) The mass spectra were recorded on a
C-H
C-H
MALDI-TOF mass spectrometer (Bruker Daltonics, Autoflex III). Monitoring the
reactions and checking the purity of the final products were carried out by thin layer
chromatography (TLC) using precoated silica gel on aluminum sheets (60 mesh
1
0

containing fluorescent indicator F , Merck) and visualization of the spots was carried
2
54
out under ultraviolet light (UV) at 365 and 254 nm.
3
.2. Typical Experimental Procedures
Preparation of 4-amino-2,6-diaryl-5-cyanopyrimidines, 3a-k: An appropriate bisnitrile
[8,11]
[6,10]
2
a-k
and arylamidine 1a-k
in equimolar quantity (3.46 mmol) were dissolved in
methanol (10 mL) and refluxed for 6h in the presence of piperidine as a base. The
contents were cooled to room temperature and solvent evaporated to give a solid mass
which was crystallized and recrystallized with an appropriate solvent.
4
-Amino-5-Cyano-2-(P-Diethylaminophenyl)-6-(P-Nitrophenyl)-Pyrimidine (3f)
This compound was obtained as colorless crystals from MeOH in 92% yield, m.p. 222-
-1
2
23, R = 0.75(n-hexane-ethyl acetate 6:4); IR, KBr, γ cm : 3117 (NH2asymm.), 3039
f max
1
(NH2symm.), 2230 (
), 1602 (
); H-NMR (DMSO-d 300 MHz), δ: 8.24 (d, 2H,
6
J 9.3 Hz, H2’ and H6’); 7.97 (d, 2H, J 9.3 Hz, H2” and H6”); 7.73 (b, 2H, C2-NH ); 7.64
2
(d, 2H, J 9.3 Hz, H3” and H5”); 6.77 (d, 2H, J 9.3 Hz, H3’ and H5’) and δ 3.10 (s, 6H).
1
3
13
C-NMR (DMSO-d 300 MHz), the assignment of all C atoms are given in the text
6
above MS (MALDI TOF MS: m/z (molecular mass found = 360.3219; calculated for
+
C H N O M , mass = 360.1335).
19
16
6
3
4
-Amino-5-Cyano-2-(P-Methylphenyl)-6-(M-Nitrophenyl)-Pyrimidine (3g)
This compound was obtained as colorless crystals from MeOH in 95% yield, m.p. 241-
-
1
2
43, R = 0.55 (n-hexane-ethyl acetate-methanol 6:3:1); IR, KBr, γmax cm : 3443
f
1
1

1
(NH2asymm.), 3352 (NH2symm.), 2167(
), 1606 (
); H-NMR (DMSO-d 300
6
MHz) δ: 8.75 (t, J 2.10 Hz, 1H, H2”); 8.47-8.39 (m, 2H, H4” and H6”); 8.29 (d, J 8.10
Hz, 2H, H2’ and H6’); 8.06 (b , 2H, C2-NH ); 7.89 (t, J 8.10 Hz,1H, H5”) 7.34 (d, J
2
1
3
8
.10 Hz, 2H, H3’ and H5’); and 2.38 (s, 3H). C-NMR (DMSO-d 300 MHz), he
6
1
3
assignment of all C atoms are given in the text above MS (MALDI TOF MS: m/z
+
+
(
molecular mass found = 332.3184; calculated for C H N O [M 1] , mass =
18 14 5 2
3
32.1069).
3
.3. Sensibility Test Of The Drugs (Minimum Inhibitory Concentration - MIC)
For determining the MIC, the compounds were dissolved individually in a solution
containing 20% dimethyl-sulfoxide (DMSO) and 80% Tween-80. The culture medium
employed was Mueller Hinton Broth (Difco). The microorganisms used in the present
test were: Staphylococcus aureus UFPEDA 02; Bacillus subtilis UFPEDA 86 and
Pseudomonas aeruginosas UFPEDA 416 from the culture collection of the Department
of Antibiotics, Federal University of Pernambuco, all maintained in Mueller Hinton
agar. The MIC test was carried out using the micro dilution method in micro 96-well
plates containing 100 mL Mueller Hinton Broth, according to the procedure
[
12]
recommended by Standard Clinical Laboratory Institute SCLI The bacterial
suspensions were taken in the sterilized distilled water and the turbidity adjusted to 0.5 on
8
the McFarland scale (1.5x10 UFC/mL). The concentrations of all eleven compounds
tested were from 3.2 to 0.0125 mg/mL. Oxacillin and ciprofloxacine antibiotics were
[
13]
employed as standards, having the same concentrations as recommended by CLSI
Overall, 12 wells (columns) were used: In the first one, the broth; in the second, solvent
1
2

and the broth. From columns 3 through 11, in 1.0 mL of the solvent, the broth was
employed as follows : 3.2 mg/mL, 1.6 mg/mL, 0.8mg/mL, 0.4/mL, 0.2mg/mL,
0
.1mg/mL, 0.05 mg/mL, 0.025 mg/mL, and 0.0125mg/mL. In the 12th well, both broth
and bacteria were added. In the second to eleventh wells, 20mL of bacteria were added.
o
The plates were incubated at 35 C for 24 hours. When the plate cultivation period ended,
a reading was taken with naked eyes and afterwards a sterilized aqueous solution of
resazurine (0.1%) was added. After 4 hours of incubation, the reading was taken again.
Resazurine facilitates the verification of microbial proliferation, and a blue color
indicates that there is no bacterial proliferation, while a red color indicates the presence
of living cells in the process of proliferation. Thus it was possible to determine the
minimum concentration responsible for the inhibition of the micro-organisms.
Supporting Information: Full experimental details and spectroscopic data (IR spectra,
1
13
H-NMR , C-NMR , and MALDI-TOF MS) for compounds 3a-k including HSQC and
HMBC spectra for compounds 3a, and HSQC, HMBC and COSY spectra for
compounds 3d, 3f-g and 3k can be found in the ‘‘Supplementary Content’’ section of this
article’s webpage.
ACKNOWLEDGEMENTS
Our thanks are due to Pernambuco State’s Scientific and Research Foundation (FACEPE)
for providing financial assistance.
REFERENCES
1
3

[
[
[
1] Lagoja, I.M. Chem. Biodiversity. 2005, 2, 1-50.
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3] Boutaleb-Charki, S.; Marin, C.; Maldonado, C.R.; Rosales, M.J.; Urbano, J.;
Guitierrez-Sánchez, R.; Quirós, M.; Salas, J.M. Sánchez-Moreno. M. Drug Metab. Lett.
009, 3, 35-44 .
4] Ruisi, G.; Canfora, L.; Bruno, G.; Rotondo, A.; Mastropietro, T.F.; Debbia, E.A.;
Girasolo, M.A.; Megna, B. J. Organomet. Chem. 2010,695, 546-551.
5] El-Gendy, M.M.A.; Shaaban, M.; Shaaben, k.a.; El-Bondkly, A.M.; Laatsch, H. J.
Antibiot. 2008, 61, 149-157.
6] Falcão, E.P.S.; Melo, S.J.; Srivastava, R.M.; Catanho, M.T.J.A.; Nascimento, S.C.
Eur. J. Med. Chem. 2006, 41, 276-282.
2
[
[
[
[
[
7]Sharma, V.; Chitranschi, N.; Agarwal, A.K. Int. J. Med Chem. 2014, 1-31.
8] Melo, S. J. DE; SANTOS, L.C.; Falcão, E.P.S.; Srivastava, R. M. Luu-Duc, C. J.
Chem. Res., 2002, 5, 216-217.
9] Xavier, A. L.; Simas, A. M.; Falcão, Emerson P.S., Dos Anjos, J. V. Tetrahedron
Lett, 2013, 54, 3462-3465.
10] Dos Santos. L. C. Síntese de 4-amino-pirimidinas e avaliação de suas propriedades
[
[
biológicas. Dissertação, Mestrado em Ciências Farmacêuticas.- Universidade Federal de
Pernambuco, Recife, PE, Brazil. 2000.
[
11]
Mendonca Junior, F. J. B. ; Dos Anjos, J. V. ; Falcao, E. P. S. ; Melo, S. J. ;
Srivastava, R.M. Heterocycl. Commun., England, 2005, 11, 479-484.
1
4

[
12] Clinical Laboratory Standards Institute (CLSI). Methods for dilution antimicrobial
susceptibility test for bacteria that grow aerobically, seventh edition. Approved standard
M7-A8. CLSI, Wayne, PA, 2009.
[
13] Clinical Laboratory Standards Institute. Performance standards for antimicrobial
susceptibility testing: twentieth Informational Supplement. Approved M100-S20. CLSI,
Wayne, PA, 2010.
1
5

Table S1. HMBC correlations of compound of 4-Amino-5-cyano-2-(p-
diethylaminophenyl)-6-(p-nitrophenyl)-pyrimidine (3f)
1
13
H NMR signals
C NMR signals
δH 8.24 ppm (H2’ and H6’ nuclei)
δC 164.2 ppm (C2 nucleus)
δC 152.6 ppm (C4’ nucleus)
δC 130.1 ppm (C2’ and C6’ nuclei)
δC 166.5 ppm (C6 nucleus)
δH 7.97 ppm (H2’’ and H6’’ nuclei)
δC 135.5 ppm (C4’’ nucleus)
δC 130.4 ppm (C2’’ and C6’’ nuclei)
δC 128.5 ppm (C3’’ and C5’’ nuclei)
δC 135.7 ppm (C1’’ nucleus)
δH 7.64 ppm (H3’’ and H5’’ nuclei)
δH 6.77 ppm (H3’ and H5’ nuclei)
δC 130.4 ppm (C2’’ and C6’’ nuclei)
δC 128.5 ppm (C3’’ and C5’’ nuclei)
δC 123.1 ppm (C1’ nucleus)
δC 111.1 ppm (C3’ an C5’ nuclei)
1
6

Table S2. Antibacterial property of compounds 3a-k, MIC = 3.2 a 0.2 mg/mL
Sample
3a
3b
3c
3d
3e
3f
3g
3h
3i
3j
3k
Control (DMSO)
Staphylococcus
aureus
0
0
0
0
0
0
0
0
0
0
0
0.8 >3. >3. >3. 0.2 0.2 0.4 >3. >3. >3. 0.1
2
2
2
2
2
2
Bacillus subtillis
0.8 >3. >3. >3. 1.6 0.4 0.2 >3. >3. >3. 0.2
2
2
2
2
2
2
Pseudomonas
aeruginosa
0.8 >3. 3.2 >3. 0.8 0.4 >3. >3. >3. >3. 0.4
2
2
2
2
2
2
1
7

Figure S1. 4-Amino-5-cyano-2-(p-methylphenyl)-6-(m-nitrophenyl)-pyrimidine (3g)
1
8

Scheme S1. Synthesis of compounds 3a-k from 1a-k and 2a-k.
1
9

Scheme S2. Mechanism of formation of 4-amino-2,6-diaryl-5-cyanopyrimidines from
arylamidine and 2-cyanocinnamonitrille
2
0