New 1-(3-nitrophenyl)-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepines: Synthesis and computational study

Article abstract of DOI:10.3390/molecules20045392

Triazole derivatives constitute an important group of heterocyclic compounds have have been the subject of extensive study in the recent past. These compounds have shown a wide range of biological and pharmacological activities. In this work, new fused tricyclic 1-(3-nitrophenyl)-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]-benzodiazepines have been synthesized by the thermal cyclization of N′-(2,3-dihydro-1H-1,5-benzodiazepin-4-yl)-3-nitrobenzohydrazides. After screening ethanol, toluene and 1-butanol as solvents, butanol-1 was found to be the best choice for the cyclization reaction in order to obtain the highest yields of tricyclic derivatives. The chemical structures of the synthesized compounds were elucidated by the analysis of their IR, ¹H- and ¹³C-NMR spectral data. For tentative rationalization of the reaction processes, the global and local reactivity indices of certain compounds, taking part in the reaction pathway, were assessed by means of quantum mechanical calculations using the conceptual density functional theory (DFT) approach. This work could be useful for the synthesis of new heterocyclic compounds bearing a fused triazole ring.

Full text of DOI:10.3390/molecules20045392

Molecules 2015, 20, 5392-5408; doi:10.3390/molecules20045392
OPEN ACCESS
molecules
ISSN 1420-3049
www.mdpi.com/journal/molecules
Article
New 1-(3-Nitrophenyl)-5,6-dihydro-4H-[1,2,4]triazolo[4,3-
a][1,5]benzodiazepines: Synthesis and Computational Study
Lidija Kosychova 1,2,*, Antanas Karalius 3, Zita Staniulytė 1, Romualdas Aleksas Sirutkaitis 1,
Algirdas Palaima 1, Audrius Laurynėnas 1 and Žilvinas Anusevičius 1
1
Institute of Biochemistry, Vilnius University, Mokslininku 12, Vilnius LT-08662, Lithuania;
E-Mails: zita.staniulyte@bchi.vu.lt (Z.S.); romualdas.sirutkaitis@bchi.vu.lt (R.A.S.);
algirdas.palaima@bchi.vu.lt (A.P.); audrius.laurynenas@gmail.com (A.L.);
zilvinas.anusevicius@bchi.vu.lt (Z.A.)
2
Department of Technological Processes, Klaipeda University, H. Manto 84,
Klaipeda LT-91001, Lithuania
3
Royal Institute of Technology, Department of Chemistry, Teknikringen 36, Stockholm S-10044,
Sweden; E-Mail:antanas.karalius@gmail.com
* Author to whom correspondence should be addressed; E-Mail: lidija.kosychova@bchi.vu.lt;
Tel.: +370-5272-9061; Fax: +370-5272-9196.
Academic Editor: Derek J. McPhee
Received: 6 January 2015 / Accepted: 20 March 2015 / Published: 26 March 2015
Abstract: Triazole derivatives constitute an important group of heterocyclic compounds
have have been the subject of extensive study in the recent past. These compounds have
shown a wide range of biological and pharmacological activities. In this work, new fused
tricyclic 1-(3-nitrophenyl)-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]-benzodiazepines have
been synthesized by the thermal cyclization of N'-(2,3-dihydro-1H-1,5-benzodiazepin-4-yl)-3-
nitrobenzohydrazides. After screening ethanol, toluene and 1-butanol as solvents, butanol-1
was found to be the best choice for the cyclization reaction in order to obtain the highest
yields of tricyclic derivatives. The chemical structures of the synthesized compounds were
1
elucidated by the analysis of their IR, H- and 13C-NMR spectral data. For tentative
rationalization of the reaction processes, the global and local reactivity indices of certain
compounds, taking part in the reaction pathway, were assessed by means of quantum
mechanical calculations using the conceptual density functional theory (DFT) approach.
This work could be useful for the synthesis of new heterocyclic compounds bearing a fused
triazole ring.

Molecules 2015, 20
Keywords: [1,2,4]triazolo[4,3-a][1,5]-benzodiazepine; 3-nitrobenzohydrazide; five-membered
5393
fused heterocycles; quantum mechanical calculation; density functional theory (DFT); DFT
global descriptors; DFT local descriptors; Fukui index
1. Introduction
Our literature survey revealed that triazole derivatives belonging to an important group of
heterocyclic compounds have been the subject of extensive study in the recent past. The 1,2,4-triazole
nucleus has been incorporated into a wide variety of therapeutically interesting compounds associated
with diverse antibacterial, antifungal, anti-inflammatory and antihypertensive activities [1–6]. Fused
heterocyclic triazoles also possess important clinical applications. Several compounds containing
1,2,4-triazole rings are well known as non-steroidal compounds used for treatment of cancer, some of
them showing an anticonvulsant, antitumor and antiviral activity [7–11]. The chemistry of
1,5-benzodiazepines and their fused heterocyclic derivatives has received considerable attention owing
to their synthetic importance and a broad spectrum of useful properties. It is known that the activity of
fused [1,2,4]triazolo[4,3-a][1,4]benzodiazepines could be qualitatively modified by substitution at C-1
atom [12]. Among the various 1,2,4-triazoles molecules designed and synthesized in recent years,
nitro-substituted arene 1,2,4-triazole derivatives possess biological activities as well as high energetic
and anti-corrosion qualities [13–16].
Keeping these observations in mind and in continuation of our research on the synthesis of
different nitrogen-containing heterocyclic rings, annulated onto the basic 1,5-benzodiazepine bicyclic
system [17–22], in this work, we have carried out the synthesis of some new 1-(3-nitrophenyl)-5,6-
dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepine derivatives. For tentative rationalization of the
reaction outcome, the global and local reactivity indices of certain compounds taking part in the synthesis
pathway were assessed by means of the B3LYP functional method within a framework of the conceptual
density functional theory (DFT) approach.
2. Results and Discussion
2.1. Chemistry
The synthesis pathway for constructing 1-(3-nitrophenyl)-5,6-dihydro-4H-[1,2,4]triazolo[4,3-
a][1,5]benzodiazepines 4a–f is shown in Scheme 1. The precursors of thiolactams 1a–f were previously
described by us [22–24]. Compounds 1a–f were converted into the thioethers 2a–f by phase-transfer
catalyzed alkylation with an excess of iodomethane. The thioethers were synthesized to avoid the lack
of reactivity of the corresponding thiolactams toward nucleophiles. The compounds 2a,b,d were
reported previously [25,26], while compounds 2c,e,f have now been obtained in good yields for the first
time according to this procedure, The structures of the compounds were supported by the analysis of
1
13
their IR, H- and C-NMR spectra. The IR spectra exhibited typical C=N stretching bands between
1598–1592 cm−1 as well as C=O stretching bands at 1679–1650 cm−1. In the 1H-NMR spectra, signals

Molecules 2015, 20
5394
derived from the methylthio group were recorded as singlets at 2.40–2.48 ppm. This group resonated at
12.9–13.2 ppm in the 13C-NMR spectrum.
H
N
S
S
CH3
N
R
R1
H
R2
i
R
R
1a - 4a CH3
CH2C6H5
N
N
1b - 4b
1c - 4c CH3
1d - 4d
1e - 4e CH3
1f - 4f
H
CH3 CH2C6H5
C(O)CH3
CH3 C(O)CH3
R1
R1
R2
R2
H
H
1a-f
2a-f
from 2a
i, iii
H
CH3
C(O)NHC6H5
C(O)NHC6H5
from 2b-f
H
ii
NO2
H
O
N
HN
iii
N
N
N
N
R
NO2
R
N
R1
R2
N
R1
R2
3b-f
4a-f
i: CH3I, benzyltriethylammonium chloride, C6H6, 40% NaOH, rt
ii: 3-nitrobenzohydrazide, C2H5OH, 45 oC
iii: BuOH, reflux
Scheme 1. Synthesis of 1-(3-nitrophenyl)-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepines.
The reaction of thioethers 2a–f with freshly recrystallized 3-nitrobenzohydrazide in anhydrous
ethanol gave N'-(2,3-dihydro-1H-1,5-benzodiazepin-4-yl)-3-nitrobenzohydrazides 3a–f. Monitoring by
TLC, it has been found that the optimal reaction temperature was about 45 °C, and the reaction time was
18–26 h. Poorly soluble non-cyclic products 3a–f formed a precipitate, whereas the excess or unreacted
starting material remained in solution. The TLC monitoring showed that under these conditions only one
new product was obtained. Compounds 3b–f were isolated as crystalline substances and fully identified.
As reaction product 3a decomposed or partial cyclized the crude material was used as for further cyclization.
The structure of compounds 3b–f was confirmed on the basis of their spectroscopic characteristics.
The IR spectra showed the appearance of the NO2 vibration in 1531–1528 cm−1 and 1352–1347 cm−1.
The IR spectrum of 3b exhibited a sharp peak at 1671 cm−1 due to the C=O absorption. Meanwhile, in
1
the H-NMR spectra of 3b–d, f, the signals derived from the NH groups were recorded as singlet at
8.85–8.98 and 10.54–10.64 ppm and increased the number of protons in the aromatic region.
The thermal condensation of compounds 3a–f was performed after a number of experiments in order
to select the best conditions for the cyclization reactions to obtain the highest yields of the expected
tricyclic derivatives. Suspensions of N'-(2,3-dihydro-1H-1,5-benzodiazepin-4-yl)-3-nitro-benzohydrazides
3a–f were heated at reflux in different solvents. After screening ethanol, toluene and 1-butanol as
solvents, it was found that the desired product 4e was obtained only by refluxing in 1-butanol and that
this solvent was thus the best choice for the reaction, because the reaction proceeded well in a high
boiling solvent. The results are outlined in Table 1.

Molecules 2015, 20
5395
Table 1. Synthesis of 1-(3-nitrophenyl)-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]
benzodiazepines 4a–f in different solvents.
Compound
Ethanol, Time (h) Toluene, Time (h) 1-Butanol, Time (h)
4a
4b
4c
4d
4e
4f
36
43
41
48
–
19
21
20
25
‒
9
11
14
16
21
15
47
22
Generally, it was demonstrated that cyclization of the compounds proceeded within different time
periods for each compound and depended on its structure, as well as on the solubility in an appropriate
solvent. The 1-(3-nitrophenyl)-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepines 4a–f were
obtained in good yields (65%–85%) and identified by their 1H spectra which shows the disappearance
of the NH groups at 8.85–8.98 ppm and 10.54–10.64 ppm. The 13C-NMR resonances of the C-1 atom of
the triazole ring were observed at 150.2–150.6 ppm. In addition, the IR spectra of compounds 4a and 4b
displayed no signals originating from the CO group.
Recently we have described the preparation of substituted 5,6-dihydro-4H-[1,2,4]triazolo[4,3-
a][1,5]benzodiazepine derivatives from the corresponding hydrazides [26]. It was found that the reaction
of 1-acetyl-2-methyl-4-(methylsulfanyl)-2,3-dihydro-1H-1,5-benzodiazepine 2d with benzo-hydrazide
in anhydrous ethanol at room temperature gave N'-(1-acetyl-2-methyl-2,3-dihydro-1H-1,5-
benzodiazepin-4-yl)benzohydrazide in good yield, and the cyclization of this compound by refluxing in
anhydrous ethanol gave 6-acetyl-5-methyl-1-phenyl-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]-
benzodiazepine. Similar reactions with 3-nitrobenzohydrazide which are outlined in Table 2 were found
to occur only at higher temperature and much more slowly, implying that the presence of a nitro group
on the aromatic ring deactivates 3-nitrobenzohydrazide as a nucleophile.
Table 2. Favorable reaction conditions for the synthesis of compounds.
Compound
Solvent Temp (°C ) t (h)
Yield (%)
N'-(1-acetyl-2-methyl-2,3-dihydro-1H-1,5-
benzodiazepin-4-yl)benzohydrazide
ethanol
room
16
18
8
91
N'-(1-acetyl-2-methyl-2,3-dihydro-1H-1,5-
benzodiazepin-4-yl)-3-nitrobenzohydrazide
6-acetyl-5-methyl-1-phenyl-5,6-dihydro-4H-
[1,2,4]triazolo[4,3-a][1,5]benzodiazepine
6-acetyl-5-methyl-1-(3-nitrophenyl)-5,6-dihydro-
4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepine
ethanol
45
65
77
85
ethanol
78
1-butanol
118
20
2.2. Computational Study
To tentatively rationalize the interaction of 1-acetyl-2-methyl-4(methylsulfanyl)-2,3-dihydro-1H-1,5-
benzodiazepine (thioether compound 2d) with benzohydrazide and 3-nitrobenzohydrazide (Scheme 1,
synthesis pathway ii), as well as to provide a partial explanation for the cyclization reactions of
N'-(1-acetyl-2-methyl-2,3-dihydro-1H-1,5-benzodiazepin-4-yl)benzohydrazide and N'-(1-acetyl-2-

Molecules 2015, 20
5396
methyl-2,3-dihydro-1H-1,5-benzodiazepin-4-yl)-3-nitrobenzohydrazide (Scheme 1, synthesis pathway
iii), a quantum mechanical computation study of the compounds was performed within the framework
of the density functional theory (DFT) approach ([27–30] and references therein). Within the DFT
framework, the estimated global and local reactivity indices of the compounds, reflecting their
electrophilic/nucleophilic powers, can be used to assess a priori the reactivity of chemical species
involved in a chemical process at the ground state of reagents ([31], and references therein). The global
electrophilic index (ϖ) of the molecules, representing their global electrophilic power, is defined using
the following expression, ϖ = µ2/(2η) [28], where µ and η are the electronic chemical potential and the
chemical hardness of the compounds, respectively. In accordance with Koopmans’ approximation for
the closed shell-molecules, the quantities µ and η can be approached in terms of the lowest unoccupied
molecular orbital (LUMO) and the highest occupied molecular orbital (HOMO) energy values, applying
the finite difference approximation scheme, as µ ≈ (ELUMO + EHOMO)/2 and η ≈ (ELUMO − EHOMO)/2 ([27–30],
and references therein). The relative global nucleophilicity index (N) of the reactive compounds,
reflecting their global nucleophilic power, can be simply expressed in terms of their negative values of
ionization potentials (IPs) or in terms of the negative HOMO energy values as a crude approximation of
IPs, i.e., the high (low) nucleophilicity of the compounds can be associated with their low (high) IP or
EHOMO ([32–34], and references therein). In our study, the relative global N index values were obtained
by means of the HOMO energy values, using the expression, N = EHOMO(Nu) − EHOMO(TCE) [32–35], where
tetracyanoethylene (TCE), representing the lowest HOMO energy among a long series of organic
molecules ([32], references therein), was used as reference. This definition leads to positive N index
values. While the global electrophilic or nucleophilic indices reflect the extensive reactivity property of
the molecules, their local reactivity indices, defined in terms of Fukui function (FF) values, can be used
to predict which site of the molecule is most likely to be reactive. The power of the FF approach lies in
the fact that the regioselectivity of the reactions can be assessed without a modeling the chemical reaction
itself. FF can be usefully employed to identify the most probable reactive sites within a molecule, as
well as to predict intramolecular reactivity, or to measure how the reactive site(s) of molecules can be
affected by the introduction of electron-donating or -withdrawing group(s). In this work, the electrophilic
(F+ ) and nucleophilic (F− ) FF values, condensed to the particular k atoms of molecules, were assessed
k
k
by means of the single point frontier molecular orbital (FMO) approach, applying the LUMO and the
HOMO eigenvectors, respectively, and the matrix of the overlap integrals [36] (more computational
details of FF values are given in the Experimental section). By contrast to most frequently used
finite-difference approximation techniques, often leading to negative FF values, the single-point FMO
approach yields the positive FF values ([36,37], and references therein).
As shown in Figure 1a, the absolute LUMO density (|LUMO|) of thioether compound 2d and the
highest electrophilic FF value (F+ ) is mainly condensed upon the carbon atom of the thioether molecule
k
as the most probable electrophilic site, participating in the interaction with benzohydrazide or
3-nitrobenzohydrazide as nucleophiles (Scheme 1, synthesis pathway (ii) and resulting in the substitution
of methylsulfanyl group by both benzohydrazides via the nucleophilic substitution reaction mechanism.
As it has been described in literature [27], benzohydrazide molecules may undergo keto-enol
tautomerism, which may determine their nucleophilic power (Scheme 2).

Molecules 2015, 20
5397
|LUMO|
low
high
(a)
|HOMO|
low
high
(b1)
(b2)
Figure 1. (a) |LUMO| map and electrophilic Fukui function (F+ ) value of thioether
k
compound 2d; (b1) |HOMO| map and nucleophilic Fukui index (F− ) values of keto- and
k
enol- form of benzohydrazide; (b2) |HOMO| map and F− values of keto- and enol-form of
k
3-nitro-benzohydrazide. The global reactivity indices of the compounds: electrophilicity
index (ϖ) and nucleophilicity index (N). |LUMO| and |HOMO| izo-values = 0.02.

Molecules 2015, 20
5398
Scheme 2. Keto-enol tautomeric equilibria for benzohydrazide molecules.
The calculations showed that the absolute HOMO density (|HOMO|) of the keto forms of
benzohydrazide and 3-nitrobenzohydrazide as nucleophiles resides mainly upon their hydrazide moiety
(Figure 1b1,b2); benzohydrazide (Figure 1b1) exhibits the lower global ϖ value and the higher global N
value than those of 3-nitrobenzohydrazide (Figure 1b2). This implies that 3-nitro benzohydrazide
possesses a lower propensity to act as a nucleophile in the reaction with thioether compound 2d as an
electrophile. In addition, the local nucleophilicity index (Nk) values of the keto form of the nucleophile
were assessed at O1, N1 and N2 atoms of the hydrazide moiety of both hydrazides. The Nk values were
obtained by multiplying global nucleophilic (N) and local nucleophilic (F− ) indices. The calculation
k
yielded NO1 = 0.362 eV, NN1 = 0.847 eV and NN2 = 0.677 eV for benzohydrazide, and NO1 = 0.319 eV,
NN1 = 0.692 eV and NN2 = 0.684 eV for 3-nitrobenzohydrazide. These data show that the highest local
nucleophilic power of the compounds is condensed on the N1 atom of hydrazide moiety. Nevertheless,
by contrast to the computation data, the experimental results suggested that the terminal N2 atom of
hydrazides is the nucleophilic site interacting with the thioether compound 2d. This enables us to propose
that in these reactions, the enol form of benzohydrazide compound might be a proximate nucleophile.
Note that the participation of the enol form of benzohydrazide, as a more reactive nucleophile compared
to its keto form, has previously been proposed by Campodonico et al. [38], who examined the reactivity
of benzohydrazides in acetylation reactions. Based upon DFT computational and experimental results,
they suggested that benzohydrazide as a nucleophile, interacting with nitrophenyl acetate as an
electrophile, may establish an intramolecular proton rearrangement, and the enol form of the
benzohydrazide may be the active species for a nucleophilic attack. In our study, the assessment of the
global ϖ and N index values of benzohydrazide and 3-nitro-benzohydrazide (Figure 1b1,b2) showed that
the global nucleophilic powers of the enol forms of the compounds are markedly higher as compared
with the nucleophilic powers of their keto forms. The local reactivity Nk values at O1, N1 and N2 sites of
enol forms were calculated as 0.599 eV, 0.661 eV and 1.335 eV, respectively, for benzohydrazide, and
as 0.229 eV, 0.533 eV and 1.233 eV, respectively, for 3-nitrobenzohydrazide. These data show that the
highest local nucleophilicity of the compounds is condensed on the terminal N2 atom of the hydrazide
moiety which is expected to be the most probable nucleophilic site for a nucleophilic attack, and the
presence of a nitro group in the benzene ring of benzohydrazide markedly lowers its global nucleophilic
power, as well as decreases the nucleophilic power of the local sites of hydrazide moiety.
Finally, we have attempted to tentatively rationalize the impact of the nitro group on the cyclization
process of the intermediates N'-(1-acetyl-2-methyl-2,3-dihydro-1H-1,5-benzodiazepin-4-yl)benzo-hydrazide
and N'-(1-acetyl-2-methyl-2,3-dihydro-1H-1,5-benzodiazepin-4-yl)-3-nitrobenzohydrazide into triazoles
(Scheme 1, synthesis pathway iii). The |HOMO| and |LUMO| density maps of N'-(1,5-benzodiazepin-4-

Molecules 2015, 20
5399
yl)benzohydrazide and N'-(1,5-benzodiazepin-4-yl)-3-nitro benzohydrazide as well as their F− and F+
k
k
values for their nucleophilic and electrophilic reactive sites, respectively, are shown in Figure 2a,b.
|HOMO|
|LUMO|
low
high
low
high
(a)
(b)
Figure 2. (a) The |HOMO| and |LUMO| maps of N’-(1-acetyl-2-methyl-2,3-dihydro-1H-1,5-
benzodiazepin-4-yl)benzohydrazide and their highest nucleophilic (F− ) and electrophilic
k
(F+ ) Fukui function values; (b) |HOMO| and |LUMO| maps of N'-(1-acetyl-2-methyl-2,3-
k
dihydro-1H-1,5-benzodiazepin-4-yl)-3-nitrobenzohydrazide and their highest F− and F+
k
k
values. |HOMO| and |LUMO| izo-values = 0.02.
The highest |HOMO| density and the highest F− values of N'-(1,5-benzodiazepin-4-yl)benzohydrazide
k
intermediate (Figure 2a) as well as of N'-(1,5-benzodiazepin-4-yl)-3-nitro benzohydrazide intermediate
(Figure 2b) reside mainly on the N1 and N2 atoms of the diazepine and hydrazide fragments, respectively,
as nucleophilic sites. The higher F− values of the N1 atom of the diazepine fragment imply that this
k
atom is the most plausible for the interaction with the carbonyl group of the hydrazide moiety. The
|LUMO| density as well as the highest F+ value for N'-(1,5-benzodiazepin-4-yl)benzohydrazide
k
(Figure 2a) are localized mainly on the carbon atom of the C=O group of the benzohydrazide as the most
probable electrophilic site. Meanwhile, the obtained low |LUMO| density as well as low F+ function
k
value for the carbon atom site of C=O group in N'-(1,5-benzodiazepin-4-yl)-3-nitro benzohydrazide
(Figure 2b) clearly show that the presence of a nitro group in a benzene ring markedly lowers the local
electrophilic power of the carbon site in a carbonyl group. This may be one of the main factors creating
unfavorable conditions for a cyclization reaction. The synthesis of a broader set of triazole structure
compounds, the assessment of their biological activity and analyses of quantitative structure-activity

Molecules 2015, 20
5400
relationship (QSAR) of the compounds in order to define possible global and local reactivity descriptors
of the compounds, responsible for their biological activity, will be the subject of our forthcoming works.
3. Experimental Section
3.1. General Information
Melting points were determinate on a Barnstead International MEL-TEMP capillary melting point
apparatus and were not corrected. Elemental analyses (C, H, N, S) were performed on an Elemental
Analyzer CE-440. IR spectra (4000–400 cm−1) were recorded on Perkin Elmer Spectrum GX FT-IR
spectrometer in KBr pellets. 1H- and 13C-NMR spectra were recorded at 300 and 75 MHz, respectively,
on an Varian Unity Inova 300 spectrometer with DMSO-d6 (compounds 3b–f) and CDCl3 (compounds
2c,e,f and 4a–f) as solvent. The chemical shifts were referenced to tetramethylsilane (δ 1H = 0 ppm) and
the solvent signal CDCl3 (δ 13C = 77.0 ppm) and DMSO-d6 (δ 13C = 39.5 ppm). The CH3, CH2, CH and
C groups in 13C-NMR were differentiated by means of the APT or DEPT method. The reactions were
monitored by TLC using TLC Silica gel 60 F254 (Merck) plates in systems: (a) benzene–methanol
(v/v, 6:1) (compounds 2c,e,f); (b) chloroform–ethyl acetate–methanol (v/v, 14:7:1,5) (compounds 3a–f);
(c) n-butanol–acetic acid–water (v/v, 4:2:1) (compounds 4a–f). Visualization was made with UV light
(254 nm) and with iodine vapor. Thiolactams 1a–f were synthesized according to the described
procedure [22–24]. 4-(Methylsulfanyl)-2,3-dihydro-1H-1,5-benzodiazepines 2a,b,d were synthesized
from thiolactams 1a,b,d by treatment with methyl iodide [25,26].
3.1.1. General Procedure for the Synthesis of 4-(Methylsulfanyl)-2,3-dihydro-1H-1,5-
benzodiazepines 2c,e,f
A mixture of the appropriate tetrahydro-1H-1,5-benzodiazepine-2-thione 1c,e,f (0.01 mol) in benzene
(50 mL), benzyltriethylammonium chloride (1.48 g, 0.0065 mol), methyl iodide (2.5 mL, 0.04 mol) and
40% NaOH (10 mL) was stirred at room temperature. After the completion of the reaction (5–8 h, by
TLC monitoring) the filtrate was diluted with chloroform (100 mL) and water (100 mL). The organic
layer was separated and aqueous phase extracted twice with 25 mL of chloroform. The combined organic
phases were dried (Na2SO4) and evaporated to dryness in vacuum and the resulting thick oily residue
was crystallized from ethanol to give the pure target compounds.
1-Acetyl-3-methyl-4-(methylsulfanyl)-2,3-dihydro-1H-1,5-benzodiazepine (2c). Yield 87%; m.p.
105–107 °C; IR ν: 1650 (C=O), 1598 (C=N) cm−1; 1H-NMR (CDCl3) δ: 1.20 (d, 3H, J = 6.9 Hz, CH3),
1.81 (s, 3H, CH3), 2.40 (s, 3H, CH3), 2.98 (m, 1H, CH), 3.54 (dd, 1H, J = 6.0 and 12.6 Hz, CH2), 4.68
(dd, 1H, J = 12.9 and 12.6 Hz, CH2), 7.08–7.39 (m, 4H, Ar-H) ppm; 13C-NMR (CDCl3) δ: 12.7 (3-CH3),
13.1 (4-CH3 ), 22.6 (1-CH3 ), 37.8 (C-3), 59.3 (C-2), 124.5 (CH), 124.6 (CH), 128.3 (CH), 128.9 (CH),
131.4 (C), 146.7 (C), 170.6 (CO), 177.7 (=C-S) ppm. Anal. Calcd. for C13H16N 2OS (248.35): C 62.87,
H 6.49, N 11.28, S 12.91. Found: C 62.92, H 6.47, N 11.30, S 12.87.
3-Methyl-4-(methylsulfanyl)-N-phenyl-2,3-dihydro-1H-1,5-benzodiazepine-1-carboxamide (2e). Yield
80%; m.p. 140–141 °C; IR ν: 3300 (NH), 1662 (C=O), 1596 (C=N) cm−1; 1H-NMR (CDCl3) δ: 1.22 (d,
3H, J = 6.7 Hz, CH3), 2.44 (s, 3H, S-CH3), 3.02 (m, 1H, CH), 3.79 (br.s, 1H, CH2), 4.43 (br.s, 1H, CH2),

Molecules 2015, 20
5401
6.18 (s, 1H, NH), 6.90–7.49 (m, 9H, Ar-H) ppm; 13C-NMR (CDCl3) δ: 12.9 (4-CH3), 13.4 (3-CH3), 38.3
(C-3), 59.8 (C-2), 119.0 (2 × CH), 122.9 (CH), 125.2 (CH), 125.7 (C), 128.3 (CH), 128.7 (2 × CH),
129.1 (CH), 129.9 (CH), 138.6 (C), 147.5 (C), 153.9 (CO), 178.4 (=C-S) ppm. Anal. Calcd. for
C18H19N3OS (325.43): C 66.43, H 5.88, N 12.91, S 9.85. Found: C 66.57, H 5.79, N 12.98, S 9.81.
2-Methyl-4-(methylsulfanyl)-N-phenyl-2,3-dihydro-1H-1,5-benzodiazepine-1-carboxamide (2f). Yield
86%; m.p. 125–127 °C; IR ν: 3302 (NH), 1679 (C=O), 1592 (C=N) cm−1; 1H-NMR (CDCl3) δ: 1.29 (d,
3H, J = 6.3 Hz, CH3), 2.35–2.46 (m, 2H, CH2), 2.48 (s, 3H, S-CH3), 5.32 (m, 1H, CH), 6.06 (s, 1H, NH),
13
6.98–7.49 (m, 9H, Ar-H) ppm; C-NMR (CDCl3) δ: 13.2 (4-CH3 ), 19.4 (2-CH3), 41.3 (C-3), 59.71
(C-2), 118.9 (2 × CH), 122.7 (CH), 125.3 (CH), 125.6 (CH), 127.8 (C), 128.6 (2 × CH), 129.4 (CH),
130.1 (CH), 138.6 (C), 148.3 (C), 153.4 (CO), 173.6 (=C-S) ppm. Anal. Calcd. for C18H19N3OS (325.43):
C 66.43, H 5.88, N 12.91, S 9.85. Found: C 66.51, H 5.81, N 12.96, S 9.77.
3.1.2. General Procedure for the Synthesis of N'-(2,3-Dihydro-1H-1,5-benzodiazepin-4-yl)-3-nitro-
benzohydrazides 3b–f
A mixture of 3-nitrobenzohydrazide (2g, 0.011 mol) and 4-(methylsulfanyl)-2,3-dihydro-1H-1,5-
benzodiazepine 2b–f (0.007 mol) in dry ethanol (50 mL) was stirred at 45 °C. After the completion of
the reaction (18–26 h, by TLC monitoring), the mixture was concentrated under reduced pressure to a
volume of 30 mL. After cooling, the mixture was kept in a refrigerator overnight, and the precipitate was
collected and recrystallized from an ethanol to give the pure compounds 3b–f.
N'-(1-Benzyl-2-methyl-2,3-dihydro-1H-1,5-benzodiazepin-4-yl)-3-nitrobenzohydrazide (3b). Yield 71%;
m.p. 169–171 °C; IR: ν 3180 (NH), 3120 (NH), 1670 (CO), 1594 (C=N), 1528 (NO2), 1347 (NO2) cm−1;
1H-NMR (DMSO-d6) δ: 1.05 (3H, d, J = 6.3Hz, CH3), 2.7-4.0 (3H, m, CH2CH), 4.30 and 4.43 (2H, AB-q,
J = 14.8 Hz, CH2Ph), 6.95–8.90 (14H, m, Ar-H, NH), 10.54 (1H, s, NH) ppm; 13C-NMR (DMSO-d6) δ:
15.0 (2-CH3), 40.9 (C-3), 53.2 (5-CH2), 59.5 (C-4), 122.1 (CH), 123.0 (CH), 123.5 (CH), 126.7 (CH),
126.9 (CH), 127.7 (2 × CH), 128.0 (C), 128.1 (2 × CH), 129.4, 129.6, 134.2, 134.4, 134.5, 135.6 (C),
138.5(C), 147.6 (C-NO2), 155.9 (C=N), 163.7 (2-CO) ppm. Anal. Calcd. for C24H23N5O3 (429.47): C
67.12, H 5.40, N 16.31. Found: C 67.24, H 5.31, N 16.23.
N'-(1-Acetyl-3-methyl-2,3-dihydro-1H-1,5-benzodiazepin-4-yl)-3-nitrobenzohydrazide (3c). Yield 85%;
1
m.p. 245–247 °C; IR: 3205 (NH), 1669 (CO), 1600 (C=N), 1526 (NO2), 1350 (NO2) cm−1; H-NMR
(DMSO-d6) δ: 1.44 (3H, d, J = 6.7 Hz, CH3), 1.84 (3H, s, CH3), 2.96 (1H, m, CH), 3.61 (1H, dd, J = 6.4
and J = 12.7 Hz, CH2), 4.38 (1H, dd, J = 12.7 and 12.9 Hz, CH2), 7.12– 8.40 (8H, m, Ar-H), 8.85 (1H,
s, NH), 10.55 (1H, s, NH) ppm; 13C-NMR (DMSO-d6) δ: 13.1 (3-CH3), 22.6 (1-CH3), 28.3 (C-3), 54.4
(C-2), 123.0 (CH), 124.6 (CH), 126.1 (CH), 127.9 (CH), 129.8 (CH), 130.2 (CH), 130.8 (CH), 131.2 (C),
131.4 (C), 134.3 (CH), 134.9 (C), 147.7 (C-NO2 ), 150.4 (C=N), 156.3 (CO), 169.2 (1-CO) ppm. Anal.
Calcd. for C19H19N5O4 (381.39): C 59.84, H 5.02, N 18.36. Found: C 59.93, H 4.97, N 18.21.
N'-(1-Acetyl-2-methyl-2,3-dihydro-1H-1,5-benzodiazepin-4-yl)-3-nitrobenzohydrazide (3d). Yield 65%;
m.p. 249–251 °C; IR ν: 3206 (NH), 1669 (CO), 1587 (C=N), 1528 (NO2), 1349 (NO2) cm−1; 1H-NMR
(DMSO-d6) δ: 1.11 (3H, d, 3J = 6.6 Hz, CH3), 1.64 (3H, s, CH3), 2.59 (1H, m, CH2), 5.01 (1H, m, CH),

Molecules 2015, 20
5402
7.17–8.76 (8H, m, Ar-H), 8.98 (1H, br.s, NH), 10.59 (1H, br.s, NH) ppm; 13C-NMR (DMSO-d6) δ: 18.7
(2-CH3), 22.7 (1-CH3), 37.2 (C-3), 53.3 (C-2), 122.0 (CH), 122.3 (CH), 124.5 (CH), 124.9 (CH), 125.6
(CH), 129.2 (CH), 129.7 (CH), 130.6 (C), 131.0 (C), 134.1 (CH), 137.9 (C), 147.6 (C-NO2), 153.6
(C=N), 160.8 (CO), 168.7 (1-CO) ppm. Anal. Calcd. for C19H19N5O4 (381.39): C 59.84, H 5.02, N 18.36.
Found: C 60.01, H 4.94, N 18.28.
3-Methyl-4-[2-(3-nitrobenzoyl)hydrazino]-N-phenyl-2,3-dihydro-1H-1,5-benzodiazepine-1-carboxamide
(3e). Yield 62%; m.p. 230–232 °C; IR ν: 3302 (NH), 3071 (NH), 1681 (CO), 1594 (C=N), 1530 (NO2),
1352 (NO2) cm−1; 1H-NMR (DMSO-d6) δ: 1.45 (3H, br.d, CH3), 2.92 (1H, m, CH), 3.83 (1H, br.s, CH2),
4.19 (1H, br.s, CH2), 6.92–8.72 (16H, m, 3NH, Ar-H) ppm; 13C-NMR (DMSO-d6) δ: 13.1 (3-CH3), 28.9
(C-3), 56.5 (C-2), 121.1 (2 × CH), 122.9 (CH), 123.2 (CH), 124.6 (CH), 126.0 (CH), 128.1 (CH), 128.3
(2 × CH), 129.2 (CH), 130.4 (CH), 130.5 (CH), 131.4 (C), 132.7 (C), 134.5 (CH), 135.1 (C), 139.3
(C-NH), 148.0 (C-NO2), 150.6 (C), 155.0 (CO), 156.5 (CO) ppm. Anal. Calcd. for C24H22N6O4 (458.47):
C 62.87, H 4.84, N 18.33. Found: C 62.91, H 4.77, N 18.25.
2-Methyl-4-[2-(3-nitrobenzoyl)hydrazino]-N-phenyl-2,3-dihydro-1H-1,5-benzodiazepine-1-carboxamide
(3f). Yield 59%; m.p. 197–200 °C; IR ν: 3424 (NH), 3063 (NH), 1684 (CO), 1594 (C=N), 1531 (NO2),
1349 (NO2) cm−1; 1H-NMR (DMSO-d6) δ: 1.20 (3H, d, J = 6.1Hz, CH3), 1.99–2.75 (2H, m, CH2), 4.86
13
(1H, m, CH), 6.8–8.8 (14H, m, NH, Ar-H), 8.96 (1H, s, NH), 10.64 (1H, s, NH) ppm; C-NMR
(DMSO-d6) δ: 19.1 (2-CH3), 37.6 (C-3), 54.5 (C-2), 120.1 (2 × CH), 122.3 (CH), 123.1 (CH), 124.7
(CH), 124.8 (CH), 125.6 (CH), 128.2 (2 × CH), 128.8 (CH), 129.1 (CH), 129.8 (CH), 131.1 (C), 132.6
(C), 134.1 (CH), 138.5 (C), 139.6 (C-NH), 147.6 (C-NO2), 153.6 (C=N), 158.7 (CO), 161.1 (CO) ppm.
Anal. Calcd. for C24H22N 6O4 (458.47): C 62.87, H 4.84, N 18.33. Found: C 62.94, H 4.81, N 18.23.
3.1.3. Synthesis of 6-Benzyl-4-methyl-1-(3-nitrophenyl-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]
benzodiazepine (4a)
The mixture of 3-nitrobenzohydrazide (2 g, 0.011 mol) and 4-(methylsulfanyl)-2,3-dihydro-1H-1,5-
benzodiazepine (2a, 2.08g. 0.007 mol) in dry ethanol (50 mL) was stirred at 45 °C temperature. After
the completion of the reaction (18 h, by TLC monitoring) the mixture was filtered, a solid residue was
dissolved in 50 mL of butanol-1 and was heated 9 h, under reflux. The solution was concentrated under
reduced pressure to a volume of 20 mL. After cooling the precipitate was collected and recrystallized from
ethyl acetate to give the pure compound 4a. Yield 71%; m.p. 188–189 °C; IR ν: 1602 (C=N), 1541
(NO2), 1348 (NO2) cm−1; 1H-NMR (CDCl3) δ: 1.54 (3H, d, J = 6.6Hz, CH3), 3.04 (1H, m, CH), 3.3–3.6
(2H, m, CH2), 4.23 and 4.52 (2H, AB-q, J = 14.2 Hz, CH2), 6.75–7.3 (9H, m, Ar-H), 7.51 (1H, m,
13
H-5'), 7.72 (1H, m, H-6'), 8.25 (1H, m, H-4'), 8.32 (1H, m, H-2') ppm; C-NMR (CDCl3) δ: 13.3
(4-CH3), 29.4 (C-4), 57.0 (6-CH2), 65.0 (C-5), 123.0 (CH), 123.4 (CH), 123.5 (CH), 124.3 (CH), 125.1
(CH), 127.4 (CH), 127.8 (2 × CH), 128.5 (2 × CH), 128.8 (C), 129.3 (C), 129.6 (CH), 129.7 (CH), 134.2
(CH), 137.3 (C), 142.7 (C), 148.2 (C-NO2), 150.6 (C), 157.9 (C) ppm. Anal. Calcd for C24H21N5O2
(411.46): C 70.06, H 5.14, N 17.02. Found: C 70.14, H 5.03, N 16.91.

Molecules 2015, 20
5403
3.1.4. General Procedure for the Synthesis of 1-(3-Nitrophenyl)-5,6-dihydro-4H-[1,2,4]triazolo[4,3-
a][1,5]benzodiazepines 4b–f
A suspension of N'-(2,3-dihydro-1H-1,5-benzodiazepin-4-yl)-3-nitrobenzohydrazide (0.007 mol)
3b–f in 1-butanol (50 mL) was heated under reflux. After the completion of the reaction (9–21 h, by
TLC monitoring) the solution was concentrated under reduced pressure to a volume of 20 mL. After
cooling the precipitate was collected and recrystallized from an ethyl acetate to give the pure
compounds 4b–f.
6-Benzyl-5-methyl-1-(3-nitrophenyl-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepine (4b). Yield
1
74%; m.p. 181–182 °C; IR ν: 1599 (C=N), 1541 (NO2), 1349 (NO2) cm−1. H-NMR (CDCl3) δ: 1.27
(3H, br.d, CH3), 2.41 (1H, br. s, CH2), 3.42 (1H, br.s, CH2), 4.06 (1H, m, CH), 4.23 and 4.54 (2H, br.
AB-q, CH2), 6.75–7.60 (11H, m, Ar-H), 8.22 (1H, m, H-4'), 8.30 (1H, m, H-2') ppm; 13C-NMR (CDCl3)
δ: 15.6 (5- CH3), 31.3 (C-4), 54.5 (6-CH2), 62.1 (C-5), 123.2 (CH), 124.0 (CH), 124.1 (CH), 124.7 (CH),
126.2 (C), 127.2 (CH), 127.8 (2 × CH), 128.3 (2 × CH), 128.6 (CH), 129.2 (CH), 129.4 (CH), 130.8 (C),
133.9 (CH), 137.7 (C), 139.5 (C), 148.1 (C-NO2), 150.2 (C), 154.9 (C) ppm. Anal. Calcd. for
C24H21N5O2 (411.46): C 70.06, H 5.14, N 17.02. Found: C 70.11, H 5.08, N 16.94.
6-Acetyl-4-methyl-1-(3-nitrophenyl-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepine
(4c).
Yield 78%; m.p. 247–248 °C; IR ν: 1668 (CO), 1600 (C=N), 1539 (NO2), 1353 (NO2) cm−1; 1H-NMR
(CDCl3) δ: 1.64 (3H, d, J = 6.7 Hz, CH3), 1.97 (3H, s, CH3), 2.97 (1H, m, CH), 3.66 (1H, dd, J = 12.7
and 6.3 Hz, CH2), 4.63 (1H, dd, J = 12.7 and 12.9 Hz, CH2), 6.94-8.33 (8H, m, Ar-H) ppm; 13C-NMR
(CDCl3) δ: 13.1 (4-CH3), 22.8 (6-CH3), 28.9 (C-4), 55.2 (C-5), 123.0 (CH), 124.7 (CH), 125.9 (CH),
128.1 (C), 129.8 (CH), 130.3 (2 × CH), 130.9 (CH), 131.9 (C), 134.0 (CH), 135.3 (C), 148.1 (C-NO2),
150.6 (C), 156.7 (C), 169.8 (6-CO) ppm. Anal. Calcd. for C19H17N5O2 (363.37): C 62.80, H 4.72, N
19.27. Found: C 62.71, H 4.68, N 19.16.
6-Acetyl-5-methyl-1-(3-nitrophenyl-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepine (4d). Yield
85%; m.p. 250–252 °C; IR ν: 1657 (CO), 1600 (C=N), 1534 (NO2), 1354 (NO2) cm−1; 1H-NMR (CDCl3)
δ: (two rotomer’s in ratio 90:10, in brackets—smaller) 1.32 (3H, d, J = 6.4Hz, CH3), [1.43 (3H d,
J = 6.4 Hz, CH3)], 1.89 (3H, s, CH3), [2.17 (3H, s, CH3)], 2.33 (1H, dd, J = 12.4 and 15.0 Hz, CH2),
[2.39 (1H, dd, J = 11.4 and 15.0 Hz, CH2)], 3.44 (1H, dd, J = 6.0 and 15.0 Hz, CH2), [3.49 (1H, dd,
J = 6.0 and 14.9 Hz, CH2)], [4.67 (1H, m, CH)], 5.30 (1H, m, CH), 6.90–8.66 (8H, m, Ar-H) ppm;
13C-NMR (CDCl3) δ: 19.1 (5-CH3), 23.2 (6-CH3), 30.0 (C-4), 54.7 (C-5), 122.9 (CH), 124.7 (CH), 125.7
(CH), 128.1 (C), 130.0 (CH), 130.2 (CH), 130.3 (CH), 132.4 (CH), 133.2 (C), 133.8 (CH), 134.3 (C),
148.1 (C-NO2), 150.2 (C), 153.9 (C), 169.1 (6-CO) ppm. Anal. Calcd. for C19H17N5O2 (363.37): C 62.80,
H 4.72, N 19.27. Found: C 62.81, H 4.63, N 19.19.
6-Carbamoyl-4-methyl-1-(3-nitrophenyl-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepine (4e).
Yield 73%; m.p. 239–241 °C; IR ν: 3423 (NH), 1681 (CO), 1594 (C=N), 1529 (NO2), 1349 (NO2) cm−1;
1H-NMR (CDCl3) δ: 1.64 (3H, br.d, CH3), 2.97 (1H, m, CH), 3.81, (1H, m, CH2 ), 4.55 (1H, m, CH2),
6.17 (1H, s, NH), 6.96–8.28 (13H, m, Ar-H) ppm; 13C-NMR (CDCl3) δ: 13.1 (4-CH3), 29.4 (C-4), 55.8
(C-5), 119.5 (2 × CH), 123.0 (CH), 123.8 (CH), 124.7 (CH), 126.5 (CH), 128.1 (C), 129.0 (2 × CH),

Molecules 2015, 20
5404
130.0 (CH), 130.4 (CH), 130.8 (CH), 131.1 (CH), 133.2 (C), 134.1 (C), 134.5 (CH), 137.8 (C), 147.9
(C-NO2), 150.7 (C), 153.4 (C), 156.8 (6-CO) ppm. Anal. Calcd. for C24H20N6O3 (440.45): C 65.45, H
4.58, N 19.08. Found: C 65.53, H 4.62, N 18.93.
6-Carbamoyl-5-methyl-1-(3-nitrophenyl-5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepine (4f).
Yield 65%; m.p. 222–223 °C; IR ν: 3411 (NH), 1678 (CO), 1595 (C=N), 1533 (NO2), 1350 (NO2) cm−1.
1H-NMR (CDCl3) δ: 1.42 (3H, d, J = 6.3 Hz, CH3), 2.37 (1H, dd, J = 11.6 and 14.8 Hz, CH2), 3.51, (1H,
m, J = 6.1 and 14.9 Hz, CH2), 5.32 (1H, m, CH), 6.19 (1H, s, NH), 7.00–8.30 (13H, m, Ar-H) ppm;
13C-NMR (CDCl3) δ: 19.8 (5-CH3), 30.3 (C-4), 55.2 (C-5), 119.5 (2 × CH), 122.8 (CH), 123.6 (CH),
124.3 (CH), 126.0 (CH), 127.9 (C), 128.8 (2 × CH), 130.3 (2 × CH), 130.6 (CH), 131.5 (C), 132.7 (CH),
133.5 (C), 134.2 (CH), 137.8 (C), 147.8 (C-NO2), 150.2 (C), 152.7 (C), 153.9 (6-CO) ppm. Anal. Calcd.
for C24H20N6O3 (440.45): C 65.45, H 4.58, N 19.08. Found: C 65.59, H 4.60; N 18.96.
3.2. Quantum Mechanical Computations
3.2.1. General
The quantum mechanical computation (QMC) was performed using the Spartan 10 software
package [39]. The initial refinement of the geometry of the compounds was carried out by the mechanical
force field (MMFF94) method, and further optimization was performed using semi-empirical PM6
Hamiltonian. The final optimization and computation of the compounds were performed by
DFT-B3LYP method in conjunction with 6-31G* basis set. All structures were globally optimized
without symmetry restrictions, and the stationary points of the optimized structures were verified by
frequency analysis. For all the optimized structures, no imaginary frequencies were defined.
3.2.2. The Assessment of DFT-Based Global and Local Reactivity Indices
The DFT quantities such as the global electronic chemical potential index (µ), the global chemical
hardness index (η) and the global electrophilicity index (ϖ) of molecules were assessed within a
framework of Koopman’s theory approach. The global nucleophilicity index (N) values of the
nucleophilic compounds were obtained by applying the method proposed by Domingo et al. [35].
For prediction of the local reactivity of the compounds, the Fukui function (FF) values for
electrophilic attack (F− ) and nucleophilic attack (F+ ) were assessed by the single-point frontier
k
k
molecular orbital (FMO) approach proposed by Contreras et al. [36], involving the highest unoccupied
molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) coefficients
(eigenvectors), respectively, and the matrix of the overlap integrals between the basis vectors. The
calculations were performed as follows: first, the FF values for the orbital components (α) of the atomic
orbital (µ) (Fα ) were computed as Fα = c c S = c2 + c
c S , where Cµα and Cνα are µ-th
να μν
µ

μα 
μ
μα να μα
μα
ν
ν ≠α
coefficient and ν-th expansion coefficient, respectively, and Sµν is an overlap integral between the µ and
ν orbital components; next, the FF values, condensed upon k atom of the compound (F± ), were obtained
k
F = Fα
α
by summing F values (
) and finally checked for the normalization condition ( F =1).
µ

k
μ

k
μ∋k
k
These calculations were performed by employing PYTHON open-source programming language. The

Molecules 2015, 20
5405
local Nk values condensed upon particular reactive sites of nucleophiles were obtained by multiplying
the global N index value by local F− values, Nk = N × F− .
k
k
4. Conclusions
The interaction of the 4-methylsulfanyl-1,5-benzodiazepine thioethers 2a–f with 3-nitro-benzo-
hydrazide afforded the corresponding 1,5-benzodiazepin-2-ylidene)-3-nitrobenzohydrazides 3a–f. The thermal
cyclization of 3a–f provided a simple way to obtain novel tricyclic 5,6-dihydro-4H-[1,2,4]triazolo[4,3-
a][1,5] benzodiazepine derivatives 4a–f. These reactions were found to proceed at a higher
temperature and much more slowly as compared with those when benzohydrazide was used instead of
3-nitrobenzohydrazide. To tentatively rationalize the experimental results, a preliminary computational
analysis of the compounds involved in the reaction pathway was performed within the framework of the
density functional theory (DFT) approach. The assessment of the DFT-based global and local reactivity
indices of the compounds clearly showed that the presence of a nitro group on the benzene ring of the
benzohydrazide markedly lowers the global nucleophilicity of the compound as well as decreases the
local nucleophilic power of its hydrazide moiety. Furthermore, the presence of a nitro group on the
benzene ring of the 1,5-benzodiazepin-2-ylidene-3-nitrobenzohydrazide intermediate was found to
markedly decrease the local electrophilic power of the carbon site in the carbonyl group. This may be
one of the main factors creating unfavorable conditions for a cyclization reaction. Since triazole structure
compounds are known to constitute a class of widely used antifungal agents, the synthesis of a broader
set of triazoles, the assessment of their biological activity and the QSAR analyses of possible
dependences of the biological activity of triazoles on their quantum mechanically assessed reactivity
indices will be the subject of our forthcoming study.
Acknowledgments
The authors thank Maryte Kreneviciene (Vilnius University) for the recording of all regular NMR spectra.
Author Contributions
Lidija Kosychova participated in coordination of this work, carried out the synthesis of the
compounds and spectra interpretation. Antanas Karalius, Zita Staniulytė and Algirdas Palaima carried
out the synthesis of the compounds. Romualdas Aleksas Sirutkaitis carried out spectra interpretation,
Žilvinas Anusevičius and Audrius Laurynėnas performed the quantum mechanical calculations.
Lidija Kosychova and Žilvinas Anusevičius wrote the manuscript. They all have approved the final
version of the manuscript.
Conflicts of Interest
The authors declare no conflict of interest.

Molecules 2015, 20
5406
References
1. Colanceska-Ragenovic, K.; Dimova, V.; Kakurinov, V.; Molnar, D.G.; Buzarovska, A. Synthesis,
antibacterial and antifungal activity of 4-substituted-5-aryl-1,2,4-triazoles. Molecules 2001, 6,
815–824, doi:10.3390/61000815.
2. Kucukguzel, I.; Kucukguzel, S.G.; Rollas, S.; Kiraz, M. Some 3-thioxo/alkylthio-1,2,4-triazoles
with a substituted thiourea moiety as possible antimycobacterials. Bioorg. Med. Chem. Lett. 2001,
11, 1703–1707, doi:10.1016/S0960-894X(01)00283-9.
3. Amir, M.; Shikha, K. Synthesis and anti-inflammatory, analgesic, ulcerogenic and lipid peroxi-dation
activities of some new 2-[(2,6-dichloroanilino)phenyl]acetic acid derivatives. Eur. J. Med. Chem.
2004, 39, 535–545, doi:10.1016/j.ejmech.2004.02.008.
4. Labanauskas, L.; Udrenaite, E.; Gaidelis, P.; Brukstus, A. Synthesis of 5-(2-, 3- and 4-
methoxyphenyl)-4H-1,2,4-triazole-3-thiol derivatives exhibiting anti-inflammatory activity.
Farmaco 2004, 59, 255–259, doi:10.1016/j.farmac.2003.11.002.
5. Tozkoparan, B.; Kupeli, E.; Yesilada, E.; Ertan, M. Preparation of 5-aryl-3-alkylthio-l,2,4-triazoles
and corresponding sulfones with antiinflammatory–analgesic activity. Bioorg. Med. Chem. 2007,
15, 1808–1814, doi:10.1016/j.bmc.2006.11.029.
6. Hasan, A.; Thomas, N.F.; Gapil, S. Synthesis, Characterization and Antifungal Evaluation of
5-Substituted-4-Amino-1,2,4-Triazole-3-Thioesters
doi:10.3390/molecules16021297.
Molecules
2011,
16,
1297–1309,
7. Guirado, A.; Lopez Sanchz, J.I.; Bautista, D.; Galvez, J. Synthesis and biological evaluation of
4-alkoxy-6,9-dichloro[1,2,4]triazole[4,3-a] quinoxalines as inhibitors of TNF-α and IL-6. Eur. J.
Med. Chem. 2012, 54, 87–94, doi:10.1016/j.ejmech.2012.04.035.
8. Swamy, S.N.; Priya, B.B.S.; Prabhuswamy, B.; Doreswamy, B.H.; Prasad, J.S.; Kanchugarakoppal, S.;
Rangappa, K.S. Synthesis of pharmaceutically important condensed heterocyclic 4,6-disubstituted-
1,2,4-triazolo-1,3,4-thiadiazole derivatives as antimicrobials. Eur. J. Med. Chem. 2006, 41,
531–538, doi:10.1016/j.ejmech.2005.12.009.
9. Bhat, K.S.; Poojary, B.; Prasad, D.J.; Naik, P.; Holla, B.S. Synthesis and antitumor activity studies
of some new fused 1,2,4-triazole derivatives carrying 2,4-dichloro-5-fluorophenyl moiety. Eur. J.
Med. Chem. 2009, 44, 5066–5070, doi:10.1016/j.ejmech.2009.09.010.
10. Johnston, G.A.R. Medicinal chemistry and molecular pharmacology of GABA-C receptors.
Curr. Top. Med. Chem. 2002, 2, 903–913, doi:10.2174/1568026023393453.
11. Clemons, M.; Coleman, R.E.; Verma, S. Aromatase inhibitors in the adjuvant setting: Bringing the
Gold to the standards? Cancer Treat. Rev. 2004, 30, 325–332, doi:10.1016/j.ctrv.2004.03.004.
12. Al-Soud, Y.A.; Al-Dweri, M.N.; Al-Masoudi, N.A. Synthesis, antitumor and antiviral properties of
some 1,2,4-triazole derivatives. Farmaco 2004, 59, 775–783, doi:10.1016/j.farmac.2004.05.006.
13. Hester, J.B., Jr.; von Voigtlander, P. 6-Aryl-4H-s-triazolo[4,3-a][1,4]benzodiazepines. Influence of
1-substitution on pharmacological activity. J. Med. Chem. 1979, 22, 489–496.
14. Bayrak, H.; Demirbas, A.; Bektas, H.; Alpay, K.S.; Demirbasm, N. Synthesis and antimicrobial
activities of some new 1,2,4-triazole derivatives. Turk. J. Chem. 2010, 34, 835–846,
doi:10.3906/kim-1004-570.

Molecules 2015, 20
5407
15. Varalakshmi, C.; Appa Rao, B.V. Inhibition of corrosion of copper by 5-mercapto-3-p-nitrophenyl-
1,2,4-triazole in aqueous environment. Anti-Corros. Methods Mater. 2001, 48, 171–180.
16. Kommu, N.; Ghule, V.D.; Kumar, A.S.; Sahoo, A.K. Triazole- substituted nitroarene derivatives:
Synthesis, Characterization and Energetic studies. Chem.-Asian J. 2014, 9, 166–178,
doi:10.1002/asia.201300969.
17. Puodžiūnaitė, B.; Jančienė, R.; Kosychova, L.; Stumbrevičiūtė, Z. On the synthetic way to novel
peri-annelated imidazo[1,5]benzodiazepinones as the potent non-nucleoside reverse transcriptase
inhibitors. ARKIVOC 2000, iv, 512–522, doi:0.3998/ark.5550190.0001.407.
18. Janciene, R.; Stumbreviciute, Z.; Vektariene, A.; Kosychova, L.; Klimavicius, A.; Palaima, A.;
Puodziunaite, B. Synthesis of novel annelated systems based on the interaction and reactivity
estimation of amino-1,5-benzodiazepin-2-ones with dimethyl-2-oxogluconate. J. Heterocycl. Chem.
2009, 46, 1339–1345, doi:10.1002/jhet.226.
19. Jančienė, R.; Stumbrevičiutė, Z.; Vektariene, A.; Kosychova, L.; Sirutkaitis, R.; Palaima, A.;
Staniulytė, Z.; Puodziunaitė, B.D. Researches on thiazolobenzodiazepines: Behavior of tetrahydro-1,5-
benzodiazepine-thiones with aromatic alpha-haloketones. Heteroatom. Chem. 2008, 19, 72–81,
doi:10.1002/hc.20414.
20. Kosychova, L.; Stumbrevičiūtė, Z.; Plečkaitienė, L.; Jančienė, R.; Puodžiūnaitė, B.D. Synthesis of
substituted 5,6-dihydro-4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepines. Khim. Geterotsikl. Soedin.
2004, 6, 943–948.
21. Kosychova, L.; Stumbrevičiūtė, Z.; Jančienė, R.; Staniulytė, Z.; Puodžiūnaitė, B.D. A convenient
synthesis of novel [1,2,4]oxadiazolo[4,3-a][1,5]benzodiazepine derivatives. ARKIVOC 2011, xi,
82–91, doi:10.3998/ark.5550190.0012.b08.
22. Jančienė, R.; Vektorienė, A.; Stumbrevičiūtė, Z.; Kosychova, L.; Klimavičius, A.; Puodžiūnaitė, B.
A Straightforward Synthesis or Novel 4H-Thiazolo-[3,2-d][1,5]benzodiazepine Derivatives.
Heteroatom. Chem. 2004, 15, 363–368, doi:10.1002/hc.20026.
23. Puodžiūnaitė, B.; Kosychova, L.; Jančienė, R.; Stumbrevičiūtė, Z. 2,3-Dihydro-1H-1,5-
benzodiazepines: A Conversion of Thiolactams to Amidines. Monatsh. Chem. 1997, 128,
1275–1281, doi:10.1007/BF00807260.
24. Janciene, R.; Stumbreviciute, Z.; Podeniene, D.; Puodziunaite, B.D.; Black, S.; Husbands, S.M.
Synthesis of novel 1-substituted [1,3]thiazolo[3,2-a]–[1,5]benzodiazepine derivatives from
1,5–benzodiazepine-2-thiones and α-halogen carbonyl coumpounds. J. Heterocycl. Chem. 2009, 43,
979–984, doi:10.1002/jhet.5570430424.
25. Puodžiūnaitė, B.; Kosychova, L.; Jančienė, R.; Stumbrevičiūtė, Z. Synthesis and mild conversion
of 1,5-benzodiazepine iminothioethers into hydrazides. Khim. Geterotsikl. Soedin. 1998, 3, 368–372.
26. Kosychova, L.; Stumbrevičiūtė, Z.; Jančienė, R.; Klimavičius, A.; Staniulytė, Z.; Palaima, A.;
Puodžiūnaitė; B.D. Synthesis of new [1,2,4]triazolo[4,3-a][1,5]benzodiazepine derivatives.
Chemija (Vilnius) 2011, 22, 60–64.
27. Pearson, R.G. Absolute electronegativity and hardness correlated with molecular orbital theory.
Proc. Natl. Acad. Sci. USA 1986, 83, 8440–8441. Available online: http://www.pnas.org/content/
83/22/8440.full.pdf (accessed on 7 August 2014).
28. Parr, R.G.; Szentpaly, L.; Liu, S. Electrophilicity Index. J. Am. Chem. Soc. 1999, 121, 1922–1924,
doi:10.1021/ja983494x.

Molecules 2015, 20
5408
29. Pearson, R.G. Chemical hardness and density functional theory. J. Chem. Sci. 2005, 117, 369–377.
Available online: http://www.ias.ac.in/chemsci/Pdf-sep2005/369.pdf (accessed on 8 August 2014).
30. Geerlings, P.; de Proft, F.; Langeneker, W. Conceptual density functional theory. Chem. Rev. 2003,
103, 1793–1873.
31. Domingo, L.R.; Perez, P.; Seez, J. Understanding C-C bond formation in polar reactions. An
ELF analysis of the Friedel-Crafts reaction between indoles and nitroolefins. RCS Adv. 2013, 3,
7520–7528, doi:10.1039/c3ra40929e.
32. Perez, P.; Domingo, L.R.; Duque-Norena, M.; Chamorro, E. A condensed-to-atom nucleophilicity
index. An application to the direct effects on the electrophilic aromatic substitutions. J. Mol.
Struct.-Theochem. 2009, 895, 86–91.
33. Contreras, R.; Andres, J.; Safont, V.S.; Campodonico, P.; Santos, J.G. A theoretical study on the
relationship between nucleophilicity and ionization potentials in solution phase. J. Phys. Chem. A
2003, 107, 5588–5593, doi:10.1021/jp0302865.
34. Domingo, L.R.; Chamorro, E.; Perez, P. Understanding the reactivity of captodative ethylenes in
polar cycloaddition reactions. A theoretical study. J. Org. Chem. 2008, 73, 4615–4624,
doi:10.1021/jo800572a.
35. Domingo, L.R.; Perez, P. The nucleophilicity N index in organic chemistry. Org. Biomol. Chem.
2011, 9, 7168–7175, doi:10.1039/C1OB05856H.
36. Contreras, R.R.; Fuentealba, P.; Galvin, M.; Perez, P. A direct evaluation of regional Fukui functions
in molecules. Chem. Phys. Lett. 1999, 304, 405–413, doi:10.1016/S0009-2614(99)00325-5.
37. Fuentealba, P.; Perez, P.; Contreras, R. On the condensed Fukui function. J. Chem. Phys. 2000, 113,
2544–2551, doi:10.1063/1.1305879.
38. Campodonico, P.R.; Aliaga, M.E.; Santos, J.G.; Castro, E.A.; Contreras, R. Reactivity of
benzohydrazide derivatives towards acetylation reaction. Experimental and theoretical studies.
Chem. Phys. Lett. 2010, 488, 86–89, doi:10.1016/j.cplett.2010.01.052.
39. Spartan’10, Version 1.1.0.; Wavefunction, Inc.: Irvine, CA, USA, 2010. Available online:
http://www.wavefun.com (accessed on 22 March 2011).
Sample Availability: Samples of the compounds 4a–f are available from the authors.
© 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article
distributed under the terms and conditions of the Creative Commons Attribution license
(http://creativecommons.org/licenses/by/4.0/).