Bioorganic Chemistry 80 (2018) 43–56

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Bioorganic Chemistry
journal homepage: www.elsevier.com/locate/bioorg

Cytoprotective and antioxidant properties of organic selenides for the T

myelin-forming cells, oligodendrocytes
⁎
Saad Shaabana,b,c,f, , Dominique Vervandier-Fasseurb, Pierre Andreolettia, Amira Zarrouka,d,
Philippe Richardb, Amr Negme, Georg Manolikakesf,g, Claus Jacobh,
⁎⁎
Mustapha Cherkaoui-Malkia,
a
Univ. Bourgogne-Franche Comté, Laboratoire BioPeroxIL (Biochimie du Peroxysome, Inflammation et Métabolisme Lipidique) EA 7270, 21000 Dijon, France
b
Institut de Chimie Moléculaire de l'Université de Bourgogne, UMR6302, CNRS, Université Bourgogne Franche-Comté, F-21000 Dijon, France
c
Organic Chemistry Division, Department of Chemistry, Faculty of Science, Mansoura University, El-Gomhorya Street, 35516 Mansoura, Egypt
d
Faculté de Médecine, Laboratoire de Nutrition-Aliments Fonctionnels et Santé Vasculaire (LR12ES05), Monastir & Faculté de Médecine, Université de Monastir, Sousse,
Tunisia
e
Biochemistry Division, Department of Chemistry, Faculty of Science, Mansoura University, El-Gomhorya Street, 35516 Mansoura, Egypt
f
Institute of Organic Chemistry and Chemical Biology, Goethe-University Frankfurt, Max-von-Laue-Str. 7, 60438, Frankfurt/Main, Germany
g
Department of Chemistry, TU Kaiserslautern, Erwin-Schrödinger-Str. Geb. 54, D-67663 Kaiserslautern, Germany
h
Division of Bioorganic Chemistry, School of Pharmacy, Saarland University, Campus B2 1, D-66123 Saarbuecken, Germany

A R T I C LE I N FO A B S T R A C T

Keywords: Here a new series of twenty-one organoselenides, of potential protective activity, were synthesized and tested for
Organic selenides their intrinsic cytotoxicity, anti-apoptotic and antioxidant capacities in oligodendrocytes. Most of the organo-
Apoptosis selenides were able to decrease the ROS levels, revealing antioxidant properties. Compounds 5b and 7b showed
Neurodegeneration a high glutathione peroxidase (GPx)-like activities, which were 1.5 folds more active than ebselen. Remarkably,
Oligodendrocytes
compound 5a diminished the formation of the oligodendrocytes SubG1 peak in a concentration-dependent
Multicomponent reactions
Cytoprotective
manner, indicating its anti-apoptotic properties. Furthermore, based on the SwissADME web interface, we
performed an in-silico structure-activity relationship to explore the drug-likeness of these organoselenides,
predicting the pharmacokinetic parameters for compounds of interest that could cross the blood-brain barrier.
Collectively, we present new organoselenide compounds with cytoprotective and antioxidant properties that can
be considered as promising drug candidates for myelin diseases.

1. Introduction Oligodendrocytes support the long-term neurons integrity by myeli-

In the brain, the myelin sheath plays a crucial role for neuron signal
transmission and their survival [1]. Such a sheath is built by a multi-
layered spiral extension of oligodendrocyte cell membranes to wrap the
neuron’s axons [2]. Chronic erosion of this myelin sheath leads to de-
myelination, and disrupting the neuron integrity and the normal axonal
function [3]. Demyelination is largely associated with numerous neu-
rodegenerative diseases (e.g., Parkinson’ and Alzheimer’s diseases) as
well as multiple sclerosis and peroxisomal leukodystrophies rare dis-
eases [4,5]. Thus for the myelination process, oligodendrocyte integrity
is central and their dysfunction was also detected in several psychiatric
disorders, including autism, schizophrenia, and depression [6].
nating their axons [7]. One of the hallmarks of oligodendrocyte dys-
function and apoptosis is their high susceptibility to oxidative stress
(OS), which results from the excess production of reactive oxygen
species (ROS; e.g., hydrogen peroxide, superoxide, and hydroxyl radi-
cals), by cellular aerobic metabolism, particularly from mitochondrial
respiration [8]. Oxidative damage, caused in a variety of neurodegen-
erative diseases, is mediated by excessive production of ROS, which can
be linked to cell lysis or oxidative burst associated to the immune re-
sponse [9]. This overproduction occurs when the generation of reactive
oxygen species (ROS) exceeds the antioxidant cellular capacity, in-
cluding enzymatic (catalase, glutathione peroxidase and superoxide
dismutase) and non-enzymatic defense systems (glutathione and

Abbreviations: CNS, Central nervous system; MTT, 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; DCF, 7-ketocholesterol (7kc), dichlorofluorescein; DHE, dihy-
droethidium; GPx, glutathione peroxidase; ROS, reactive oxygen species; DPPH, 2,2-diphenyl-1-picrylhydrazyl; ABTS, 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid
⁎
Corresponding author at: Organic Chemistry Division, Department of Chemistry, Faculty of Science, Mansoura University, El-Gomhorya Street, 35516 Mansoura, Egypt.
⁎⁎
Corresponding author at: Univ. Bourgogne-Franche Comté, Laboratoire BioPeroxIL EA7270, Faculté des Sciences Gabriel, 6 Bd Gabriel, 21000 Dijon, France.
E-mail addresses: dr_saad_chem@mans.edu.eg (S. Shaaban), malki@u-bourgogne.fr (M. Cherkaoui-Malki).

https://doi.org/10.1016/j.bioorg.2018.05.019
Received 31 March 2018; Received in revised form 16 May 2018; Accepted 20 May 2018
Available online 21 May 2018
0045-2068/ © 2018 Elsevier Inc. All rights reserved.
S. Shaaban et al.
ascorbic acid) [10]. The selenoprotein family, including glutathione
peroxidases, represents one of the first-line defenses against ROS. Se-
lenoproteins play also a crucial role for neuronal function, as demon-
strated by deactivation of the tRNA [Ser] (Sec) gene in mice, resulting
in extensive neurodegeneration [11]. The second line of defense is re-
presented by cellular antioxidants, such as glutathione (GSH) and se-
lenium (Se), the latter for its implication in the active center of several
glutathione peroxidases and in the catalytic activity of thioredoxin re-
ductase [12]. Depleted states of the reduced form of GSH and selenium
were associated with the progression of neurodegenerative diseases
[13–15]. This underlines the overarching importance of an essential
dietary supply of Se for mammals. Interestingly, there seems to be a
connection between Se and other antioxidants on the one side, and the
role of ROS in the pathogenesis of neurodegenerative diseases on the
other [16–18]. Yeo et al., reported that Se antioxidant activity pre-
vented cell apoptosis in an experimental rat spinal cord injury [19].
Numerous data revealed that Se levels decline in humans with age and
that Se might play different roles in the evolution of Alzheimer’s disease
[20,21]. Se deficiency leads to inhibition of the normal upregulation of
myelin genes in differentiating oligodendrocytes and the progression
from immature to mature oligodendrocytes as well [22]. Curiously,
experiments using animal models reveal that organoselenium has a
better bioavailability and biological activity than inorganic Se [23].
Therefore, during the last decade or two, the biological chemistry of
selenium derivatives has often focused on potential antioxidant and
neuroprotective activities [24], and here primarily on organic rather
than inorganic selenium agents [25,26]. During our research on sele-
nium derivatives with possible antioxidant and neuroprotective activ-
ities, we recently developed organoselenium compounds with enhanced
antioxidant and GPx-like activities [27–32]. The mechanism by which
these compounds exert their antioxidant effect is related to the mod-
ulation of the ROS and GSH levels in cancer cells. Interestingly, some of
these compounds showed better antioxidant and GPx-like activities
than classical antioxidants (e.g., vitamin C, ebselen) [27,30,32–36].
Based on these results, we decided to further direct our research to
obtain evidences regarding the structural requirements underlying the
potential cytoprotective/neuroprotective activities of organic selenides
taking the myelin-forming cells, oligodendrocytes as a model of study.
Different reports on the synthesis of organoselenium compounds
have already been published; however, they generally require the
handling of hazard reagents (e.g., KSeCN), harsh reaction conditions
(e.g., strongly acidic or basic), and mostly multistep procedures
[37,38]. Therefore, we decided to avoid rather sophisticated synthetic
procedures and employ simple, straight forward and facile routes to-
wards the synthesis of our preliminary organoselenium-based library.
Here, we present the results obtained during extensive in vitro
evaluations of the selenium derivatives in oligodendrocytes. Besides a
rational design of synthetic organoselenium compounds, we have
aimed at gaining a deeper insight into the potential use of these com-
pounds as neuroprotectors in neurodegenerative diseases through the
protective effect on oligodendrocytes, the myelinating cells. For the
chemical synthesis, we adopted both multistep and one-pot synthesis as
part of a diversity-oriented approach to achieve the synthesis of the
twenty-one organoselenides. The in vitro evaluations were conducted
using immortalized murine oligodendrocyte cell lines [39]. Flow cyto-
metry was used to estimate the ROS levels employing the H2-DCFDA
and the DHE assays. Furthermore, the antioxidant potential of these
compounds was further investigated employing different chemical and
biochemical assays and the apoptotic/anti-apoptotic properties of the
compounds were detected by flow cytometry follow-up of the sub-G1
peak. Finally, to assess the structure-activity relationship of the syn-
thesized organoselenium compounds, we determined in silico their
physicochemical and pharmacokinetic parameters, and their drug-
likeness as well through the free web tools in SwissADME.
44
Bioorganic Chemistry 80 (2018) 43–56
2. Results and discussions
2.1. Design and syntheses
The last decade has witnessed significant advance in the synthesis of
diverse classes of organic selenides e.g., diselenides, selenocyanates,
and seleno-heterocyclic compounds. Although, these compounds have a
pivotal role in human health and disease prevention; they have limited
application in biological systems due to their often toxicity especially if
higher concentrations were administrated. Combining selenium with
better physico-chemical properties bioactive pharmacophores will in-
teractively not only stimulate their corresponding cytotoxicity but will
also foster their bioavailability. Within this context, our design strategy
relies on the synthesis of agents in which selenium is combined with
different functionalities such as quinone, cyclic imides, N-substituted
maleanilic and succinanilic acids, formamide, acetanilide, acetaldehyde
diethyl acetal, and/or benzyl. Most of these functionalities are com-
monly found in bioactive compounds, thus enabling the chemical di-
versity and increasing polarity.
Based on our recent reports on organoselenium agents [28–35], we
envisioned the synthesis of novel selenium-based primary aromatic
amines 2a-d. The amine functionality in turn can give an access to a
large group of agents either via isocyanide-based multicomponent re-
actions (IMCR) (e.g., Passerini, azido-Ugi Passerini, Ugi, and Groebke-
Blackburn-Bienaymé reactions) or via the classical amines reactions
(e.g., formylation, acetylation, etc.).
The key synthons selenoamines 2a-d were synthesized in good
yields (up to 91%) via reduction of the diselenide (1) with sodium
tetrahydridoborate (NaBH4) followed by subsequent nucleophilic sub-
stitution (SN) reaction with appropriate organic halides (Scheme 1).
Chalcogen-bearing isonitriles are not commercially available and
usually challenging to synthesize. In 2009, we reported the synthesis of
three chalcogen-based isonitriles, including the tellurium isonitrile for
the first time [32,34]. Therefore, our preliminary target was to convert
the amines 2a, 2b, 2c, and 2d into the corresponding isonitriles, sui-
table for further applications in IMCR [29,40]. The synthesis of iso-
cyanides is usually performed in two steps i.e. amine formylation em-
ploying acetic formic anhydride followed by subsequent dehydration
using phosphoryl chloride (POCl3). In accordance with our aim, we
decided to apply the same methodology for the transformation of or-
ganoselenium-based amines 2a-d into the corresponding isocyanides.
Unexpectedly, acetanilides 4a-c were invariably obtained instead of the
corresponding formamides. This promoted us to use only formic acid in
place of the acetic formic anhydride. Under neat conditions, the desired
corresponding formamides were obtained in good yields (up to 88%).
The dehydration of formamides was thereafter investigated using
POCl3. A non-polar product (eluted with petroleum ether) with the
characteristic isonitrile odor was detected by TLC; however, the triplet
13
CNMR peak (≈156 ppm) corresponding to the isocyanide carbon did
not appear, suggesting that the isocyanide may be unstable. Therefore,
we designed a one-pot synthetic approach based on the use of the in situ
formed isocyanide in subsequent isocyanide-based MCRs (IMCRs) such
as Groebke-Blackburn-Bienaymé and Ugi reactions [40]. Unfortunately,
very poor conversion (< 5%) was constantly observed with subsequent
disappearance of the isocyanide from the reaction mixture (monitored
by TLC). Hence, we concluded, that this result is related to the high
reactivity and low stability of the in situ formed isocyanides. Different
dehydrating agents such as cyanuric chloride [41], triphenylphosphine
[42], 4-toluenesulfonyl chloride [43], DABCO [44] and methyl N-
(triethylammoniumsulfonyl)carbamate [45] were investigated; how-
ever, these reagents turned out to be either low yielding or ineffective
with our seleno-formamides.
Further efforts were then directed to evaluate the reactivity of the
novel selenoamines 2a-d via their reactions with various anhydrides
aiming to synthesize selenocyclic imides which we recently proved to
possess enhanced antioxidant activities (e.g., GPx-mimicking and free
S. Shaaban et al.
Scheme 1. Synthesis of the selenoamines 2a-d, selenoformamides 3a-c and selenoacetamides 4a-c. Reagents and conditions: (i) Amine (1 mmol), NaBH4 (5 mmol);
(ii) Amine (1 mmol), 85% aq. HCOOH (1.2 eq), 2 h, 60 °C; (iii) Amine (1 mmol), HCOOH (1 eq), acetic anhydride (1.5 eq), 2 h, 55 °C.
radical-scavenging activity) [28–31]. Amines 2b-c reacted with maleic
and succinic anhydrides in dry toluene at ambient temperature afforded
the corresponding N-substituted maleanilic 5a-b and N-substituted
succinanilic 6a-b derivatives in quantitative yields (Scheme 2). In
contrast, 2a failed to react under these conditions and a tar material
was formed. This may be attributed to the side reaction(s) such as hy-
drolysis and sequential condensation and/or polymerization which
might take place at the acetal residue. Furthermore, the resulting N-
substituted monoamidic acids 5a-b and 6a were subjected to acid-cat-
alyzed esterification and the corresponding methyl esters 7a-b and 8a
were obtained in good yields (up to 87%). The esterification of 6b was
also performed under the same conditions as of 6a. Unfortunately, the
expected quinone-based methylester was not obtained; instead we ob-
served a decomposition of 6b. We attributed this behavior to the for-
mation of the corresponding hydroquinone (under acidic conditions)
and subsequent cleavage of the amidic bond.
As expected, selenocyclic imides 10a-b were obtained via ring-clo-
sure of the corresponding N-substituted succinanilic acids 6a-b with
acetic anhydride at 55 °C in the presence of sodium acetate (Scheme 2).
Unexpectedly, kinetically controlled isomaleimides 9a-b were ob-
tained in good yields (up to 77%) instead of the thermodynamically,
more stable, maleimides. This was confirmed via spectral data and X-
ray diffraction. A perspective view and atom numbering are given in
Fig. 1 and crystal structure determination and refinement parameters
are shown in the Experimental section.
The X-ray single crystal structure of 9a shows a triclinic system with
a space group of P-1. The phenyl ring is directly bonded to the imine
moiety (eN]). There are three different types of CeO bonds: C(14)eO
45
Bioorganic Chemistry 80 (2018) 43–56
(2) single bond with a bond distance of 1.389 Å, C(17)eO(2) single
bond with a bond distance of 1.405 Å, and C(17)eO(2) double bond
with a bond distance of 1.195 Å. Such efficient and clean pathway to
isomaleimides synthesis is highly useful and worth further investiga-
tions using a wider arsenal of N-substituted selenomonoamidic acids.
2.2. Cytoprotective and antioxidant profiles
2.2.1. Organoselenides effect on the viability of the oligodendrocytes 158 N
and 158JP
Oligodendrocytes play a crucial in the long-term neurons integrity
by myelinating their axons and thus securing the fast signal conduction
[2,3]. Oligodendrocytes are the most vulnerable cells of the CNS to ROS
and there are only very few reports on the cytoprotective/cytotoxic role
of organoselenides on these myelinating cells. It was therefore of in-
terest to investigate the protective activity of the synthesized organo-
selenides on two immortalized murine oligodendrocytes: 158N and
158JP cell lines. The jimpy (158JP) cell line is mutated for the pro-
teolipid protein PLP/DM20, a predominant myelin protein that may
have a role in the maintenance of myelin sheaths [39]. To determine
the half maximal inhibitory concentration IC50 of the organoselenides, a
range of concentrations was evaluated on both oligodendrocytes cell
lines. The low cytotoxic concentration will be further selected to eval-
uate the compounds corresponding antioxidant and protective proper-
ties. Accordingly, MTT assay was used to evaluate the basal oligoden-
drocytes cytotoxicity of the twenty-one organic selenides on 158N and
158JP oligodendrocytes using 7-ketocholesterol (7kc) as known cyto-
toxic positive control [46]. MTT is metabolized by the mitochondrial
S. Shaaban et al.
Scheme 2. Reactions of selenoamines 2a-d with anhydrides and subsequent dehydration/esterification. Reagents and conditions: (i) amine (1 mmol), toluene (5 ml),
anhydride (1 mmol), r.t, 3 h; (ii) amide-acid (1 mmol), MeOH (10 ml), conc.H2SO4 (200 µl), 8 h, r.t; (iii) amide-acid (1 eq), 100 mg NaOAc, Ac2O (3 ml), 2 h, 50–60 °C.
enzyme succinate dehydrogenase giving the blue formazan product.
Hence, this assay provides information on viability (and/or cell pro-
liferation) and mitochondrial metabolism. The concentration producing
50% growth inhibition (IC50, Table 1) and the maximum concentration
without toxicity were estimated for each of the compounds from the
respective dose-response curves.
As shown in Table 1, most of the compounds did not exhibit any
effect on the viability (IC50 ≥ 100 µM) of 158N and 158JP at the con-
centration range used, except for the quinoid-based compounds 6b and
10b and N-substituted maleanilic ester 7a, they showed IC50 values
46
Bioorganic Chemistry 80 (2018) 43–56
down to 9 and 3 µM in 158N cells and 158JP, respectively. Surprisingly,
the cytotoxicity of acetanilides 4a and 4c was more pronounced in the
case of 158JP cells compared to 158N cells (IC50 = 0.02 and 14 µM,
respectively). These compounds may act as pro-oxidants and their
toxicity was significant in 158JP owing to the fact that these cells have
higher ROS levels (10-fold) compared to 158N, due to the known higher
mitochondrial activity and redox status in 158JP [47]. A possible ex-
planation of acetanilides derivatives cytotoxicity, is may be related to
the accumulation of paracetamol, a well-known cytotoxin, produced
during the intracellular xenobiotic metabolism of acetanilides [48].
Fig. 1. Ortep view of of (Z)-5-((4-((4-bromophenyl)
selanyl)phenyl)imino)furan-2(5H)-one (9a). Selected
bond lengths (Å): C11-N1 1.414, C14-N1 1.266, C14-
O2 1.389, C17-O1 1.195, C17-O2 1.405. Selected
bond angles (o): C14-N1-C11 126.4, N1-C14-O2
126.1, C14-O2-C17 107.77, O1-C17-O2 120.0.
S. Shaaban et al. Bioorganic Chemistry 80 (2018) 43–56

Table 1
Newly synthesized organoselenides, yields and their effect on the viability of 158N and 158JP cells.a
R1SePhR2 R1 R2 Yields % IC50 (µM)

158N 158JP

2a eCH2CH(OC2H5)2 eNH2 89 54 -b
2b eCH2C6H5 eNH2 85 -b -b
2c eCH2-4-Br-C6H4 eNH2 91 -b -b
2d -3-Me-1,4-naphthoquinone eNH2 89 27 -b
3a eCH2CH(OC2H5)2 eNHCHO 82 25 -b
3b eCH2-4-Br-C6H4 eNHCHO 88 43 91
3c -3-Me-1,4-naphthoquinone eNHCHO 81 39 -b
4a eCH2CH(OC2H5)2 eNHCOCH3 89 14 0.02
4b -CH2C6H5 eNHCOCH3 92 -b -b
4c 3-Me-1,4-naphthoquinone eNHCOCH3 75 11 0.02
5a eCH2-4-Br-C6H4 eNHCOCH]CHCOOH 94 -b -b
5b 3-Me-1,4-naphthoquinone eNHCOCH]CHCOOH 91 -b 35
6a eCH2-4-Br-C6H4 eNHCOCH2CH2COOH 90 54 -b
6b 3-Me-1,4-naphthoquinone eNHCOCH2CH2COOH 93 26 3
7a eCH2-4-Br-C6H4 eNHCOCH]CHCOOMe 86 9 20
7b 3-Me-1,4-naphthoquinone eNHCOCH]CHCOOMe 84 -b -b
8a eCH2-4-Br-C6H4 -NHCOCH2CH2COOMe 87 -b 83
9a eCH2-4-Br-C6H4 77 29 50

9b 3-Me-1,4-naphthoquinone 75 -b -b

10a eCH2-4-Br-C6H4 76 -b -b

10b 3-Me-1,4-naphthoquinone 76 16 5

7kc – – – 37 74

a
The cytotoxicity was measured after one day of treatment with different concentrations (0, 1, 10, 20, 50 and 100 µM) of the organic selenides using the MTT
assay. The IC50 was estimated as the mean of two parallel experiments; 7kc was used as a positive control.
b
Means no growth inhibition was observed at the tested concentration range.

2.2.2. Assessment of antioxidant profiles of the selenide compounds
Organoselenium compounds are generally good nucleophiles and
this fact led to the rational design of several synthetic organoselenides
as antioxidants, chemopreventive, and neuroprotective agents
[36–38,49,50]. To the best of our knowledge, the underlying me-
chanism(s) of action is (are) not fully understood. Organoselenium
compounds can either act as pro-oxidants or antioxidants depending on
the redox status of their environment. In normal cells, organic selenides
behave as antioxidants and protect cells from ROS. In contrast, these
compounds switch into pro-oxidants in cells with elevated ROS levels
[51–54].
Taking into account that the CNS is prone to oxidative damage and
in the light of our search for new neuroprotective agents, our aim is to
further investigate the antioxidant potential of the novel organic sele-
nides employing different biochemical and chemical assays.
2.2.2.1. Effect of organoselenides on oligodendrocytes ROS
production. ROS levels were estimated by flow cytometry in 158N
cells treated with different concentrations (0, 10, 20 and 50 µM) of the
organoselenium compounds employing the commonly used H2-DCFDA
and DHE assays for the detection of ROS and of superoxide anions
(O2%−), respectively [55,56]. The commercial α-Tocopherol (vitamin
E) was used as a standard antioxidant (positive control).
In the H2-DCFDA assay, most of the compounds were able to de-
crease the ROS levels in 158N cells. Interestingly, compounds 2b, 2c,
3b, 4a, 4b, 4c, 5a, 5b, 6b, and 9a showed a significant decrease in the
H2-DCF fluorescence intensity, thus lower ROS production at 10 and
20 µM compared to vitamin E (Table 2). On the other hand, compounds
7a, 8a and 10a and most of the quinone-based organoselenium 7b, 9b,
and 10b were able to mediate significant overproduction of the
intracellular ROS levels at all the tested concentrations proposing a pro-
oxidant activity of these compounds (Table 2).
Interestingly, the pro-oxidant activity was more pronounced in the
case of the quinoid compounds 2d which showed the highest H2-DCF
fluorescence intensity. This is hardly surprising, as quinones in general
and selenium-containing naphthoquinones, in particular, are well
known to generate ROS in a catalytic fashion - and via the reduction of
dioxygen in the presence of a suitable reducing agent.
2.2.2.2. Effect of organoselenium compounds on superoxide radical anion
(O2.−) production by oligodendrocytes. In living cells and under aerobic
conditions, quinones undergo redox cycling with triplet oxygen to
generate superoxide radical anion (O2%−). The later causes oxidative
DNA damage, apoptosis and subsequent cell death [55]. This
biochemical feature is responsible for most of the quinone-based
drug’s activity. From this perspective, all the selenoquinones were
selected for further estimation of their respective ability to produce
O2%− by the DHE assay. The latter is believed to be a selective “gold
standard” O2%−detector. DHE is a lipophilic cell-permeable probe,
which is rapidly oxidized by O2%−, to give the fluorescent compound 2-
hydroxyethidium [55]. As shown in Table 3, and in agreement with the
literature and our hypothesis of quinones as ROS-generating
substances, all quinoid-based compounds were able to increase the
O2%− levels, compared to vitamin E. Interestingly, compound 2d was
among the most active compounds and was able to induce a dose-
dependent increase of O2%− in 158N cells.
2.2.3. Anti-apoptotic effect of selenide compounds on the oligodendrocytes
Apoptosis was estimated by staining fixed 158N cells with PI fol-
lowed employing flow cytometry quantification of the sub-G1 peak in

47
S. Shaaban et al. Bioorganic Chemistry 80 (2018) 43–56

Table 2 any effect on the development of the SubG1 peak (data not shown).
Evaluation of the effects of compounds on ROS production by H2-DCFDA as- Compound 5a is reducing ROS levels, which in essence is an anti-
say.a oxidative and hence often also an anti-apoptotic event. Admittedly,
10 µM Foldb 20 µM Foldb 50 µM Foldb some of the other compounds (e.g., 5b, 6b and 9a) are a lot better in
removing ROS, yet, they may not be as good in crossing membranes.
2a 71 ± 5* 0.75 81 ± 10* 0.86 82 ± 10* 0.87 This may be due to that 5a is probably amphiphilic and hence a good
2b 38 ± 4* 0.40 38 ± 5* 0.40 33 ± 2* 0.35
molecule to enter cells, whilst others are not. Compound 5a is also an
2c 36 ± 8* 0.38 41 ± 2* 0.43 48 ± 2* 0.51
2d 700 ± 143* 7.44 854 ± 135 9.08 935 ± 307* 9.94 antioxidant (albeit a weak one) and may act via its unsaturated bond.
3a 43 ± 2* 0.45 46 ± 1* 0.49 71 ± 209 0.76 There is another possible mode of action: is an attack of 5a on cysteine
3b 34 ± 6* 0.36 33 ± 5* 0.35 38 ± 8* 0.40 proteins via its unsaturated double bond, and this may result in an
3c 124 ± 7 1.32 115 ± 9 1.22 95 ± 14 1.01
upregulation of antioxidant pathways and hence anti-apoptotic action
4a 45 ± 2* 0.48 49 ± 11* 0.52 54 ± 20* 0.57
4b 20 ± 14* 0.21 15 ± 6* 0.16 13 ± 3* 0.14
as well (for instance via Nrf-2). All this is, of course, speculation, and
4c 7 ± 2* 0.07 5 ± 1* 0.05 6 ± 1* 0.06 these results worth further in-depth studies to substantiate the anti-
5a 42 ± 10* 0.45 36 ± 20* 0.38 31 ± 20* 0.32 apoptotic effect of 5a. Induction of anti-apoptotic effect in oligoden-
5b 11 ± 2* 0.12 8 ± 3* 0.09 10 ± 4* 0.11 drocytes by 5a is a key target for chemotherapeutic and chemopre-
6a 72 ± 32 0.77 64 ± 14* 0.68 51 ± 19* 0.54
ventive applications of such a compound, opening the way for further
6b 15 ± 0* 0.16 24 ± 0* 0.26 26 ± 1* 0.28
7a 127 ± 52 1.35 133 ± 50 1.42 103 ± 50 1.10 pharmacological studies.
7b 241 ± 74* 2.56 218 ± 48* 2.32 250 ± 71* 2.66
8a 127 ± 40 1.35 135 ± 47 1.44 140* ± 40 1.49 2.2.4. Glutathione peroxidase-like activity assay
9a 7 ± 4* 0.07 8 ± 4* 0.09 11 ± 4* 0.12
Many organoselenium compounds have long been known to mimic
9b 226 ± 79* 2.40 216 ± 51* 2.30 178 ± 23* 1.90
10a 179 ± 150 1.90 164 ± 100 1.74 147 ± 75* 1.60
the antioxidant activity of the GPx selenoenzyme. Among these com-
10b 217 ± 11* 2.30 192 ± 82 2.04 197 ± 54* 2.10 pounds, the non-toxic seleno-organic drug, ebselen has been extensively
Vitamine E 94 ± 5 studied because of its superior GPx mimetic activity. Recently, we have
a
reported different selenium-based organic compounds that possess
158N Cells were cultured in the presence of different compounds (C1 = 10; higher catalytic GPx-like activity than ebselen. In view of the former,
C2 = 20; C3 = 50 µM); Flow cytometry technique was used to evaluate ROS
the novel organic selenides were evaluated for their GPx mimicking
levels via staining cells with H2-DCF; data are shown as mean ± SD and re-
activity [30,60].
present the absolute values after corrections (subtraction of the untreated cells
The GPx catalyzes the reduction of ROOR, in presence of glu-
fluorescence).
b tathione, thus protecting cells from oxidative damage by H2O2 [61–63].
Fold of the compounds was calculated as follow: result assay at the given
The reaction takes place at the enzyme catalytic site, which contains
concentration/result of Vitamin E. significance of the difference between the
selenium redox center as a selenocysteine amino acid. It is worthwhile
DMSO and compounds. Treated cells are indicated by (Mann–Whitney test; *:
to mention that selenium is an essential cofactor in all the selenocys-
P < 0.05); vitamin E (50 µM) was used as a positive control.
teine-based GPx family members for the manifestation of their corre-
sponding antioxidant activities [64].
Table 3
Evaluation of the effects of compounds on superoxide anions production by
The catalytic GPx mimic activities of the newly synthesized organic-
DHE assay. selenides were evaluated employing the NADPH-reductase coupled
assays [56–68]. The latter is repeatedly used for the evaluation of the
DHE assay (% of positive cells)
GPx-like activity of organochalcogens. The assay principle depends on
Vitamine E 74 ± 1* the reduction of H2O2 by GPx followed by oxidation of glutathione
10 μM 20 μM 50 μM disulfide (GSSG) and NADPH to glutathione (GSH, reduced form) and
2d 264 ± 25* 406 ± 29* 417 ± 31* NADP+, respectively. The latter can be spectrophotometrically detected
3c 115 ± 8 109 ± 1 117 ± 6
by a decrease in the absorbance at 340 nm. The known, standard GPx
4c 109 ± 12 121 ± 1 120 ± 4
5b 102 ± 4 105 ± 4 112 ± 9 mimic compound, ebselen is used as the positive control. The back-
6b 211 ± 38* 220 ± 41* 252 ± 41* ground reaction of GSH with H2O2 was measured and the activity of the
9b 166 ± 32* 172 ± 33* 179 ± 31* GPx was calculated after correction. The reaction was followed till the
10b 133 ± 37 151 ± 28* 218 ± 40* end and the rate of the reaction was constantly linear (Fig. 3).
7b 86 ± 15 87 ± 15 91 ± 30
Interestingly, as shown in Fig. 3, the quinoid-based N-substituted
158N Cells were grown in the presence of different compounds (C1 = 10; maleanilic 5b acid and its corresponding methyl esters 7b were more
C2 = 20; C3 = 50 µM); Flow cytometry techniques was used to estimate the active (≈1.5-fold) than ebselen. The rest of the compounds showed
O2%− production via staining with DHE; Data are shown as mean ± SD and moderate GPx activity, particularly: 2d, 3b, 8a and 9b compounds.
expressed as % control; Significance of the difference between the DMSO and
compounds. Treated cells are indicated by (Mann–Whitney test; *: P < 0.05); 2.2.5. Assessment of antioxidant/pro-oxidant activity: DPPH, ABTS radical
Vitamin E (50 µM) was used as a positive control. scavenging, and bleomycin DNA damage assays
Among different assays used for the accounting of the antioxidant
the cell cycle. The sub-G1 is indicator of apoptotic cells and can be activities of organic compounds, the 2,2-diphenyl-2-picrylhydrazyl
identified by a nuclear fragmentation and/or condensation associated (DPPH) and 2,2′-azinobis-3-ethylbenzothioazoline-6-sulphonate (ABTS)
with an internucleosomal fragmentation of the DNA [57,58]. 7Kc is chemical assays are usually used to evaluate the radical scavenging
known to induce oxidative stress and trigger apoptosis in 158N murine activities of natural products, foods (e.g., vegetables, fruits and bev-
oligodendrocytes and therefore was used as apoptosis promoter and the erages) and several organoselenium compounds [27,30,69,70]. Fur-
positive control [59]. thermore, the bleomycin-induced DNA damage assay isconsidered as an
To detect apoptosis, cell cycle analysis was performed after treat- accurate method to evaluate the pro-oxidant activity of many food
ment of 158N cells with different concentrations (50, 10, 5 and 1 μM) of antioxidants and antineoplastic agents [69,70].
organoselenium compounds treated with or without 7kc. Interestingly, As presented in Fig. S1 and Table S1 (Supporting Information), most
compound 5a was able to diminish the formation of the SubG1 peak in of the compounds showed good-moderate scavenging activity com-
a concentration-dependent manner, which is an indicator of its anti- pared to vitamine C. On the other hand, selenoquinones 2d, 3c, 4c, 5b,
apoptotic properties (Fig. 2). The remaining compounds did not show 6b, 7b, 9b and 10b showed between 40 and 60% pro-oxidant activities,

48
S. Shaaban et al.
when compared to vitamin C (bleomycin-reducing activity considered
as 100%).
These results were in agreement with that obtained from DCF and
DHE assays that selenoquinones have more pro-oxidant properties
compared to the rest of organoselenium compounds. Although the
DPPH and the ABTS assays are widely used, they are only in vitro
models and do not necessarily translated into in vivo action, in parti-
cular, the ADME factors are not covered here either.
2.3. In silico structure-activity relationship
Based on the molecular structure, we explored in silico the synthe-
sized organoselenides drugability by performing a computer-aided
structure-activity prediction through the free web tools in SwissADME,
which is a user-friendly interface website (http://www.swissadme.ch)
to evaluate several physicochemical and pharmacokinetic parameters
[71]. Among these parameters showed in Fig. 4: the topological polar
surface area (TPSA) based on the summation of tabulated surface
contributions of polar fragments in the compound; the lipophilicity
represented by the n-octanol/water partition coefficient (log Po/w),
which is a key physicochemical parameter in drug discovery; the
pharmacokinetic (i.e. gastrointestinal (GI) absorption, blood-brain
barrier (BBB) permeability), and the synthetic accessibility (SA) score,
Fig. 3. GPx-like activity assay (µM min−1). The reduction of H2O2 by GPx-like activity was tracked by oxidation of glutathione disulfide (GSSG) and NADPH to
glutathione (GSH, reduced form) and NADP+, respectively. The latter was detected by a decrease in the absorbance at 340 nm.
49
Bioorganic Chemistry 80 (2018) 43–56
Fig. 2. Anti-apoptotic effect of the organoselenium
derivative 5a on oligodendrocytes. The anti-apop-
totic effect was measured by cytometry using pro-
pidium iodide. The organoselenium compound 5a
was used at different concentrations (50, 10, 5, and
1 μM) to evaluate the percentage of cells in Sub-G1
phase during the cell cycle of 158N cells treated
with/without 7kc (50 μM) for 24 h. Data shown are
mean ± SD.
which ranges from 1 (very easy synthesis) to 10 (very difficult synth-
esis). This computation of SA score was implemented by Daina et al.
[72] in SwissADME interface to assist the medicinal chemist for the
identification of potentially problematic fragments and eventual lead
optimization.
In parallel, to compare the structure-activity relationship of the
synthesized organoselenide derivatives, we summarized the obtained
experimental results as a heatmap of these compounds taking into ac-
count their chemical phylogeny (Fig. 4): where the scalable red color
indicates the negative effect, while the scalable green color reveals the
positive effect of the tested compounds. The heatmap of our experi-
mental results (Fig. 4) shows that selenoamines 2a (diethoxyethyl se-
lanyl aniline) and 2d (selanyl dihydronaphthalene, dione), derived
from diselinide as a precursor, exhibited a low or a mild cytotoxicity
respectively, while selenylaniline derivatives 2b and 2c revealed a
protective effect on oligodendrocytes. It’s noteworthy, that selenoa-
mines, as selenocystine, induce ROS production and exert their pro-
oxidant action through depletion of the cellular antioxidant glutathione
[73–75]. Interestingly, compound 2d, which show a higher cytotoxicity
of the 2-serie compounds, presents also strong pro-oxidant and DNA
damage activities and a lower antioxidant property and all 2d deriva-
tives, containing the 2-methyl-3-selanylnaphthalene-1,4-dione moiety,
conserved a significant pro-oxidant and DNA damage activities.
S. Shaaban et al. Bioorganic Chemistry 80 (2018) 43–56

Fig. 4. Chemical phylogeny heatmap and drug-likeness descriptors of the synthesized organoselinides. Chemical phylogeny was obtained taking into account the
synthesis design described for each compound in the result section and the experimental results were reported as a heatmap, in which the scalable red color indicates
the negative effect, while the scalable green color reveals the positive effect of the tested compounds respectively. The color scale was determined from the
experimental values presented in figures and tables. TPSA: topological polar surface area; log Po/w: n-octanol/water partition coefficient; GI: gastrointestinal; BBB:
blood-brain barrier.

However, among 2d derivatives, three compounds (5b, 7b and 9b)
exhibited a protective effect on oligodendrocytes and showed a GPx-
like activity, but only compound 5b revealed antioxidant and anti-
apoptotic properties (Fig. 4). Several studies reviewed by Bhabak and
Mugesh [76] described the mechanism of GPx activity of Ebselen and
related organoselenium compounds, for which the strength of Se⋯O
non-bonded interactions in the selenenyl sulfides fine-tuned the balance
between the antioxidant scavenging and the GPx activity. Additionally,
7b showed a negative predicted pharmacokinetic to cross the blood-
brain barrier. Structurally, compound 7b is an esterified derivative of
the N-substituted monoamidic acid 5b. Nonetheless, a strong correla-
tion exists between the absorbed fraction and the hydrogen bond de-
scriptors [77]. Here the physicochemical analysis in SwissADME re-
vealed that compound 7b has additional 4 rotatable bonds and one H-
bond donor, which may account for its unfavorable molecular dynamic.
Furthermore, TPSA was defined by Ertl et al. [78] as the sum of the
surface areas of polar fragments in a molecule. As documented by
Hitchcook et al. [77], compounds with PSA < 90 Å2 are considered
easily absorbable with higher capacity in penetrating cellular mem-
branes, while the preferred range is compounds with PSA < 70 Å2.
Moreover, substances with high TPSA are more prone to better interact
with hydrophobic environment of their transporter proteins, as shown
for the Multidrug resistance-associated protein 1 [79].
In line to this, compounds 5b, 6b, and 7b are either equally or
above the PSA limit (72.80 Å2, 100.54 Å2, and 89.54 Å2, respectively).
Interestingly, compounds 2c, 5a and 7a showed lower polar surface
than the recommended PSA limit (26.02 Å2, 66.40 Å2, and 55.40 Å2
respectively). Within this context, the 7a esterified derivative with a
lower TPSA (55.40 Å2) and a poor solubility, showed a strong cyto-
toxicity and a significant GPx activity (Fig. 4). While compound 5a
which derived from 2c, with a TPSA of 66.40 Å2, showed protective and
highest anti-apoptotic effects on oligodendrocytes as well as a good
antioxidant property, but has a low GPx activity. Thus, compound 5a
presents interesting protective properties for oligodendrocytes and
could become a lead compound in the development of new drug. The
pioneer work on drug-likeness was setup by Lipinski et al. [80] as a
rule-of-five to predict poor bioavailability for a drug candidate when it
contains more than 5H-bond donors, 10H-bond acceptors, a molecular
weight superior to 500 and a calculated log Po/w higher than 5. Re-
markably, all the substituted organoselenides that we synthesized show
favorable Lipinski rules (with zero violation) indicative of their like-
lihood drug-likeness to become an oral drug with respect to bioavail-
ability. Pharmacokinetically, except compounds 6b and 7b, all the
other synthesized organoselenides would potentially cross the blood-
brain barrier. The predicted synthetic accessibility scores range from
2.6 to 3.8, revealing that these compounds are easily synthesizable.
Such prediction is in agreement with the obtained good experimental
synthesis yields ranging from 77% to 91%. Collectively, the in silico and
in cellulo collected data are in favor of additional investigations as the
organotypic cerebellar slice model to study the effects of lead com-
pound on axon demyelination.

50
S. Shaaban et al.
3. Conclusion
In the present work, we synthesized, in good yields, twenty-one
novel organoselenium compounds and evaluated their potential anti-
oxidant and protective activities in silico and in cellulo on oligoden-
drocytes. It’s noteworthy that most of the organic selenides demon-
strated a cytoprotective effect in two different oligodendrocyte cell
lines, indicating that these compounds may represent chemoprotective
activity. Therefore, the potential antioxidant properties of the com-
pounds were evaluated employing different well-established biological
and chemical assays, including H2-DCFDA, DHE, DPPH, ABTS, bleo-
mycin DNA damage and GPx-like assays. Our data demonstrated that
most of the selenium-based quinones manifested a pro-oxidant activity,
whereas, the rest of the compounds demonstrated antioxidant activity.
Within this context, organoselenium compounds 2c, 2b, 3b, 4a, 4b, 4c,
5a, 5b, 6b, and 9a were able to decrease the intracellular ROS levels, as
measured by H2-DCFDA assay, in oligodendrocytes compared to vi-
tamin E. On the other hand, selenoquinones 2d, 3c, 7b, 9b and 10b
increased the ROS levels in oligodendrocytes. Furthermore, the apop-
totic/anti-apoptotic properties of the compounds were detected by flow
cytometric estimation of the sub-G1 peak in oligodendrocytes treated
with the 7kc apoptosis inducer. Interestingly, a significant anti-apop-
totic effect was observed in the case of compound 5a suggesting the
potential protective activity of this compound. Moreover, compounds
5b and 7b displayed good GPx-like (≈1.5-fold more active than eb-
selen) which in turn may also account for their protective actions. These
results were further confirmed by the ABTS and DPPH scavenging ac-
tivity and the bleomycin dependent DNA damage assay, where most of
the compounds showed good scavenging activity with respect to as-
corbic acid and protected DNA from damage; whereas, selenium-based
quinones exhibited low scavenging activity and caused DNA damage.
Together, the present results point to antioxidant properties of
some, non-cytotoxic, organic selenides resulting from their capacity to
reduce ROS generation. These compounds are considered promising
candidates and might provide a new avenue for innovative antioxidant
research into prototype drugs for neurodegenerative disorders.
4. Experimental section
4.1. Material and methods
All chemicals were obtained from Sigma. Solvents were dried prior
to use. The spectroscopic studies were measured at the “Pôle Chimie
Moléculaire” de l'Université de Bourgogne (PACSMUB)”. The mass of
the organic selenides was obtained on an Electro-Spray Ionization Mass
Spectrometry (Thermo LTQ Orbitrap XL ESI–MS) and high-resolution
mass spectra. 1H (300.13 MHz) and 13C (75.5 MHz) NMR spectra were
analyzed on Bruker 300 Avance III, spectrometers. The values of the
chemical shifts (δ) are presented in parts per million relative to tetra-
methylsilane, using deuterated solvent as an internal standard. All
chemicals were purchased from Sigma e.g., 7-ketocholesterol, and as-
corbic acid and vitamin E. More details on the spectroscopic data can be
found in the Supporting Informations. 2-methyl-3-bromo-1,4-naph-
thaoquinone was synthesized according the reported literature method
[81]. Compound 1 was synthesized according to a literature reported
method [82,83,84].
4.2. Biological assays
4.2.1. Cytotoxicity assay
Murine oligodendrocytes (158N and 158JP) were seeded at 240,000
cells per 24-well plates, and were grown using the Dulbecco’s Modified
Eagle Medium supplemented with penicillin antibiotics (1% v/v,
streptomycin) and fetal bovine serum (5% v/v) and kept at 37 °C in
atmosphere containing 5% CO2. The concentration of 7kc [57–59] and
vitamin E, and the time of treatment were chosen based on data
51
Bioorganic Chemistry 80 (2018) 43–56
obtained on 7kc treated 158N cells [85]. Organoselenium compounds
were prepared in DMSO and then diluted with DMEM up to 1 mM.
4.2.2. MTT assay
The MTT assay was carried out on 158N and 158JP cells after one
day of incubation with 7kc (50 µM) different concentrations (0, 1, 10,
20, 50 and 100 µM) of the organic selenides. In live cells, the MTT is
reduced to formazan by the succinate dehydrogenase mitochondrial
enzyme. The microplate reader Tecan Sunrise (Tecan, Lyon, France)
was used for the reading of the plates at 570 nm. The IC50 values and
the maximum concentrations without toxicity were estimated for each
of the compounds from the respective dose-response curves.
4.2.3. H2-DCFDA and DHE assays
ROS overproduction and O2%− and were detected with DCF and
DHE, respectively. Cultured cells were treated with trypsin re-sus-
pended in PBS (106 cells/mL) and kept in the dark for half hour at 37 °C
with H2-DCFDA or DHE at 10 μM and 2 μM and cells were analyzed by
flow cytometry [55,56]. The green fluorescence of 2′,7′- dichloro-
fluorescein resulting from the oxidation of H2-DCFDA was analyzed by
flow cytometry and collected through a 520/10-nm band pass filter.
DHE, a non-fluorescent compound, rapidly oxidized in ethidium under
the action of O2%− to give 2-hydroxyethidium that intercalates with
DNA giving red fluorescence of ethidium collected through a 590/10-
nm band pass filter using an excitation of 488 nm and an emission
wavelength of 575 nm [55,56]. Flomax (Partec) or FlowJo (Tree Star
Inc.) software were used for the data analysis.
4.2.4. Apoptosis status and detection of Sub-G1 cells
The analysis of the cell cycle can reveal a Sub-G1 peak, which is
representative of apoptotic cells. Cell cycle analysis was detected on
158N cells stained with Propidium Iodide (PI).
Flow cytometric analyses were measured on a Galaxy flow cyt-
ometer. Cells were prepared by re-suspension in cold ethanol (80%,
−20 °C) for 2 h. Cells were then washed with phosphate buffer, and re-
suspended in phosphate buffer (300 μl) containing PI (80 μg/mL) and
RNase (200 μg/mL) and kept for 1 h at 37 °C.PBS 2 ml was then added,
and Fluorescence of propidium iodide was recorded using a
590 ± 10 nm band filter. The percentage of cells in different cell cycle
phase was manually estimated.
4.2.5. Gpx-like activity
The GPx-kit was purchased from Biodiagnostic (Egypt) and used
according to the reported method [86]. The sample was prepared by
adding assay buffer and NADPH reagent (0.1 ml; 24 µmol Glutathione,
4.8 µmol NADPH and 12 units Glutathione reductase), and organic se-
lenides (0.01 ml, 41 µM). H2O2 (0.8 mM) was then added and the ab-
sorbance was kinetically recorded every 1 min at wavelength 340 nm
over three successive minutes. A340nm/min was calculated. Ebselen
(41 µM) was used as the positive control. For colored organoselenium
compounds, their individual absorbance was subtracted at the used
wavelength.
4.2.6. Synthesis and characterization
4.2.6.1. Synthesis of 2-methyl-3-bromo-l,4-naphthoquinone [81,84]. 2-
Methyl-1,4-naphthoquinone (11 mg, 63 mmol), anhydrous sodium
acetate (1.1 mg, 34 mmol) were mixed together in glacial acetic acid
(50 ml). Bromine (2 ml) was then added and the mixture was allowed to
settle in the dark for three days. A yellow crystalline mass formed,
which was removed. The supernatant liquid was poured on to 500 ml of
distilled water. The formed precipitate was filtered off and
recrystallized from ethanol.
4.2.6.1.1. General procedure Ib: Reduction of 1 and subsequent
modification of selenium based-amines (2a-c). Compound 1 (1 mmol)
and appropriate halo derivative (2.2 mmol) were dissolved in EtOH
(20 ml). Sodium tetrahydridoborate (189.15 mg, 5 mmol) was added
S. Shaaban et al.
portion wise over 0.5 h. Then the reaction was reflux for additional 3 h.
The organic layer was dried and evaporated under vacuum. The residue
was purified by silica gel chromatography.
4.2.6.1.2. General procedure Ia: Reduction of 1 and subsequent
modification of selenium based-amines (2d). Compound 1 (1 mmol) and
appropriate halo derivative (2.2 mmol) were dissolved in EtOAc
(20 ml). Water (20 ml) and N-Methyl-N,N,N-trioctylammonium
chloride (Aliquat 336) (45 mg, 5% mol) were then added. Sodium
tetrahydridoborate (189.15 mg, 5 mmol) was added portion wise over
1 h. Then the reaction was stirred for additional 3 h. The organic layer
was dried and evaporated under vacuum. The residue was purified by
silica gel chromatography.
4.2.6.1.3. General procedure IIa: Preparation of N-acetyl derivatives
(4a-c). A mixture of formic acid (1.0–1.2 equivalents) and acetic
anhydride (1.5–2.0 equivalents) was heated at 55 °C for 2 h and then
one equivalent of the appropriate amine (2a-d) was added. Heating was
continued for another 2 h. After complete consumption of the starting
material (as judged by TLC analysis), the reaction was cooled and
poured onto water. The formed precipitated was collected and purified
by silica gel chromatography.
4.2.6.1.4. General procedure IIb: Preparation of N-acetyl derivatives
(4a-c). Acetic anhydride (2.0 equivalents) was added to one equivalent
of the appropriate amine (2a-d) and the reaction was heated for two
hours at 60 °C. The reaction was cooled and then poured onto ice. The
formed precipitated was purified by silica gel chromatography.
4.2.6.1.5. General procedure III: Preparation of N-formyl derivatives
(3a-c). A solution of the amine (1.0 mmol) and 85% aqueous formic
acid (1.2 mmol) was heated at 60 °C for 2 h. After the disappearance of
the amine, the reaction was poured on to water and the crude N-formyl
compound formed essentially as a solid product was purified by silica
gel chromatography.
4.2.6.1.6. General procedure IV: Preparation of amide-acids (5a-b
and 6a-b). To a solution of amine (1 mmol) in dry toluene (5 ml), the
appropriate anhydride (1 mmol) was added and the mixture was stirred
at for 3 h. The amide-acid precipitate was collected and further washed
with hot-toluene and dried under vacuum. The purity of the amide-
acids was satisfactory for further use. If necessary, further purification
using silica gel chromatography was performed.
4.2.6.1.7. General procedure V: preparation of cyclic imides (10a-b)
and/or isomaleimides (9a-b). To an appropriate amide-acid (one
equivalent) and 100 mg of sodium acetate, acetic anhydride (3 ml)
was added and the mixture was gently heated for 2 h at 50–60 °C. Ice
was added and the resulting mixture was extracted with CH2Cl2
(200 ml), dried with Mg2SO4. CH2Cl2 was evaporated and the residue
was purified by silica gel chromatography.
4.2.6.1.8. General procedure VI: preparation of methyl ester (7a-b
and 8a). An appropriate amide-acid (1 mmol) was suspended in
methanol (10 ml) and then conc.H2SO4 (200 µl) was added and the
reaction was stirred at ambient temperature for 8 h. The mixture was
poured onto ice and the formed precipitate was purified by silica gel
chromatography.
4.2.6.1.9. Crystallization and X-ray data collection of 9a. Colorless
single crystals of C17H12NO2SeBr were obtained by the slow
evaporation method. A suitable crystal with dimensions of
0.37 × 0.37 × 0.12 mm3 was selected, mounted on glass fibers
and diffraction data were collected on a Bruker D8 diffractometer
using graphite monochromated MoK/a radiation (λ = 0.71073 Å).
The crystal was kept at 100 K during data collection. APEX2 [87] was
used for the unit cell determination and intensity data collection. Using
Olex2 [88], the structure was solved with the ShelXT [89] and structure
solution program using Direct Methods and refined with the XL [89]
refinement package using Least Squares minimisation. For more details
regarding the crystal parameters and data collection statistics for 9a,
please refer to the supporting information.
52
Bioorganic Chemistry 80 (2018) 43–56
4.2.6.2. 4-((2,2-Diethoxyethyl)selanyl)aniline (2a). Compound 2a was
prepared following procedure Ib from 1 (344 mg, 1 mmol),
bromoacetaldehyde diethyl acetal (331 μl, 2.2 mmol), Aliquat 336
(45 mg, 5% mol) and sodium tetrahydridoborate (189.15 mg,
5 mmol). The progress of the product formation was followed by TLC
petroleum ether: EtOAc = 6:1, Rf = 0.39, purified by column silica gel
chromatography with petroleum ether: EtOAc = 6:1.5. Colorless oil;
Yield: 258.1 mg (89%). 1H NMR (300 MHz, CDCl3) δ 7.42–7.34 (d,
J = 7.37 HZ, 2H, Ar-H), 6.62–6.55 (d, J = 6.58, 2H, Ar-H), 4.66 (t,
J = 5.7 Hz, 1H, CH), 3.67–3.46 (m, 4H, 2CH2), 3.03 (d, J = 5.7 Hz, 2H,
SeCH2), 1.19 (t, J = 7.1 Hz, 6H, 2CH3); 13C NMR (75 MHz, CDCl3) δ
146.17, 135.90, 117.11, 115.70, 102.30, 61.70, 32.23, 15.23; MS (ESI):
m/z = found 312.04 [M++Na]; calcd. 289.06 [M+]; HRMS calcd. for
C12H19NO2Se [M++Na]: 312.0498, found 312.04597 [M++Na].
4.2.6.3. 4-(Benzylselanyl)aniline (2b). Compound 2b was prepared
following procedure Ib from 1 (344 mg, 1 mmol), benzyl chloride
(253 μl, 2.2 mmol), Aliquat 336 (45 mg, 5% mol) and sodium
tetrahydridoborate (189.15 mg, 5 mmol). The progress of the product
formation was followed by TLC petroleum ether: EtOAc = 6:1,
Rf = 0.36, purified by column silica gel chromatography with
petroleum ether: EtOAc = 6:1.5. Colorless oil; Yield: 223.55 mg
(85%). 1H NMR (300 MHz, CDCl3) δ 7.28–7.18 (m, 2H, Ar-H),
7.14–7.08 (m, 2H, Ar-H), 6.94–6.81 (m, 2H, Ar-H), 6.50–6.39 (m, 2H,
Ar-H), 3.85 (s, 2H, SeCH2), 3.62 (s, 2H, NH2); 13C NMR (126 MHz,
CDCl3) δ 146.68, 138.65, 137.03, 131.32, 130.48, 120.33, 116.48,
115.63, 32.51; MS (ESI): m/z = found 263.85 [M++1]; calcd. 263.02
[M+]; HRMS calcd. for C13H13NSe [M++1]: 264.02860, found
264.02748 [M++1].
4.2.6.4. 4-((4-Bromobenzyl)selanyl)aniline (2c). Compound 2c was
prepared following procedure Ib from 1 (344 mg, 1 mmol), 4-
bromobenzyl bromide (550 mg, 2.2 mmol), Aliquat 336 (45 mg,
5% mol) and sodium tetrahydridoborate (189.15 mg, 5 mmol). The
progress of the product formation was followed by TLC petroleum
ether: EtOAc = 6:1, Rf = 0.36, purified by column silica gel
chromatography with petroleum ether: EtOAc = 6:1.5. White solid;
Yield: 310.31 mg (91%); mp 156–158 °C. 1H NMR (300 MHz, DMSO) δ
7.37–7.33 (m, 2H, Ar-H), 7.24–7.20 (m, 2H, Ar-H), 7.01–6.94 (m, 2H,
Ar-H), 6.59–6.55 (m, 2H, Ar-H), 3.90 (s, 2H, SeCH2); 13C NMR
(75 MHz, DMSO) δ 146.65, 138.63, 137.04, 131.31, 130.45, 120.32,
116.49, 115.61, 32.50; MS (ESI): m/z = found 364.84 [M++Na];
calcd. 340.93 [M+]; HRMS calcd. for C13H12BrNSe [M++1]:
341.93911, found 341.93760 [M++1].
4.2.6.5. 2-((4-Aminophenyl)selanyl)-3-methylnaphthalene-1,4-dione
(2d). Compound 2d was prepared following procedure Ia from 1
(344 mg, 1 mmol), 2-methyl-3-bromo-l,4-naphthoquinone (550 mg,
2.2 mmol), Aliquat 336 (45 mg, 5% mol) and sodium
tetrahydridoborate (189.15 mg, 5 mmol). The progress of the product
formation was followed by TLC petroleum ether: EtOAc = 4:1.5,
Rf = 0.32, purified by column silica gel chromatography with
petroleum ether: EtOAc = 3:1. Brown solid; Yield: 305.27 mg (89%);
mp 162–164 °C. 1H NMR (300 MHz, DMSO) δ 7.97 (ddt, J = 11.8, 6.9,
3.4 Hz, 2H, Ar-H), 7.87–7.77 (m, 2H, Ar-H), 7.28–7.19 (m, 2H, Ar-H),
6.55–6.44 (m, 2H, Ar-H), 5.39 (s, 2H, NH2), 1.98 (s, 3H, CH3); 13C NMR
(75 MHz, CDCl3) δ 181.84, 181.73, 149.10, 147.31, 147.24, 135.69,
134.08, 133.77, 131.69, 131.65, 126.34, 126.27, 114.65, 111.92,
16.50; MS (ESI): m/z = found 344.95 [M++1]; calcd. 343.01 [M+];
HRMS calcd. for C17H13NO2Se [M++Na]: 366.00037, found
365.99930 [M++Na].
4.2.6.6. N-(4-((2,2-Diethoxyethyl)selanyl)phenyl)formamide
(3a). Compound 3a was prepared following procedure III from 2a
S. Shaaban et al.
(289 mg, 1 mmol) and 85% aqueous formic acid (1.2 mmol). The
progress of the product formation was followed by TLC petroleum
ether: EtOAc = 8:1, Rf = 0.40, purified by column silica gel
chromatography with petroleum ether: EtOAc = 8:1.5. Brown solid;
Yield: 259.94 mg (82%); mp 175–177 °C. 1H NMR (300 MHz, CDCl3) δ
8.66 (s, 1H, CHO), 8.28 (s, 1H, NH), 7.50–7.34 (m, 3H, Ar-H),
6.96–6.88 (m, 1H, Ar-H), 4.62 (td, J = 5.6, 3.0 Hz, 1H, CH),
3.66–3.40 (m, 4H, 2CH2), 3.00 (d, J = 5.6 Hz, 2H, SeCH2), 1.12 (t,
J = 7.1 Hz, 6H, 2CH3); 13C NMR (75 MHz, CDCl3) δ 162.52, 159.18,
136.16, 135.92, 134.47, 133.84, 126.57, 125.76, 120.55, 119.28,
102.28, 62.13, 31.64, 15.22; MS (ESI): m/z = found 339.95
[M++Na]; calcd. 317.05 [M+]; HRMS calcd. for C13H19NO3Se [M+]:
317.05123, found 199.96062 [M+-CH2CH(OC2H5)2].
4.2.6.7. N-(4-((4-Bromobenzyl)selanyl)phenyl)formamide
(3b). Compound 3b was prepared following procedure III from 2c
(341 mg, 1 mmol) and 85% aqueous formic acid (1.2 mmol). The
progress of the product formation was followed by TLC petroleum
ether: EtOAc = 8:1, Rf = 0.36, purified by column silica gel
chromatography with petroleum ether: EtOAc = 6:1. White solid;
Yield: 324.72 mg (88%); mp 158–160 °C.1H NMR (300 MHz, MeOD) δ
8.47 (s, 1H, CHO), 7.56–7.38 (m, 5H, Ar-H), 7.13–6.97 (m, 3H, Ar-H),
4.11 (s, 2H, SeCH2); 13C NMR (75 MHz, MeOD) δ 167.77, 132.13,
131.46, 127.58, 127.54, 126.51, 116.38, 115.17, 28.02; MS (ESI): m/
z = found 391.86 [M++Na]; calcd. 368.93 [M+]; HRMS calcd. for
C14H12BrNOSe [M++Na]: 391.91597, found 391.91555 [M++Na].
4.2.6.8. N-(4-((3-Methyl-1,4-dioxo-1,4-dihydronaphthalen-2-yl)selanyl)
phenyl)formamide (3c). Compound 3c was prepared following
procedure III from 2d (343 mg, 1 mmol) and 85% aqueous formic
acid (1.2 mmol). The progress of the product formation was followed by
TLC petroleum ether: EtOAc = 8:1, Rf = 0.36, purified by column silica
gel chromatography with petroleum ether: EtOAc = 8:3. Brown solid;
Yield: 300.51 mg (81%); mp 168–170 °C. 1H NMR (300 MHz, DMSO) δ
10.26 (s, 1H, CHO), 8.27 (d, J = 1.9 Hz, 1H, CHO), 8.04–7.97 (m, 2H,
Ar-H), 7.85–7.80 (m, 2H, Ar-H), 7.52 (t, J = 2.5 Hz, 4H, Ar-H), 2.09 (s,
3H, CH3); 13C NMR (75 MHz, DMSO) δ 182.33, 181.52, 160.17, 149.86,
138.32, 134.57, 134.03, 132.25, 126.98, 126.78, 123.89, 120.52,
18.06; MS (ESI): m/z = found394.01 [M++Na]; calcd. 371.01 [M+];
HRMS calcd. for C18H13NO3Se [M++Na]: 393.99529, found
393.99469 [M++Na].
4.2.6.9. N-(4-((2,2-diethoxyethyl)selanyl)phenyl)acetamide
(4a). Compound 4a was prepared following procedure IIa from 2a
(289 mg, 1 mmol) and acetic anhydride (2.0 equivalents). The progress
of the product formation was followed by TLC petroleum ether:
EtOAc = 4:3, Rf = 0.30, purified by column silica gel
chromatography with petroleum ether: EtOAc = 1:1. Yellow solid;
Yield: 294.84 mg (89%); mp 165–167 °C. 1H NMR (300 MHz, CDCl3)
δ 8.37 (s, 1H, NH), 7.40–7.32 (m, 4H, Ar-H), 4.60 (t, J = 5.6 Hz, 1H,
CH), 3.63–3.37 (m, 4H, 2CH2), 3.01–2.93 (m, 2H, CH2), 2.06 (s, 3H,
COCH3), 1.10 (t, J = 7.1 Hz, 6H, 2CH3); 13C NMR (75 MHz, CDCl3) δ
169.13 (s), 137.42 (s), 133.75 (s), 124.93 (s), 120.70 (s), 102.29 (s),
62.14 (s), 31.68 (s), 24.37 (s), 15.23 (s); MS (ESI): m/z = found 376.06
[M++2Na]; calcd. 331.07 [M+]; HRMS calcd. for C14H21NO3Se [M+]:
331.0687, found 331.06812 [M+].
4.2.6.10. N-(4-(Benzylselanyl)phenyl)acetamide (4b). Compound 4b
was prepared following procedure II from 4-(benzylselanyl)aniline 2b
(263 mg, 1 mmol) and acetic anhydride (2.0 equivalents). The progress
of the product formation was followed by TLC petroleum ether:
EtOAc = 8:1, Rf = 0.42, purified by column silica gel
chromatography with petroleum ether: EtOAc = 8:1.5. White solid;
Yield: 280.6 mg (92%); mp 180–182 °C. 1H NMR (300 MHz, DMSOd6) δ
6.83–6.55 (m, 4H, Ar-H), 6.49–6.41 (m, 4H, Ar-H), 4.11 (s, 2H, SeCH2),
1.39 (s, 3H, CH3); 13C NMR (75 MHz, MeOD) δ 170.32, 139.06, 138.17,
53
Bioorganic Chemistry 80 (2018) 43–56
134.54, 128.50, 127.87, 126.31, 124.32, 120.06, 31.70, 22.42; MS
(ESI): m/z = found 327.94 [M++Na]; calcd. 305.03 [M+]; HRMS
calcd. for C15H15NOSe [M++ Na]: 328.02111, found 328.01998
[M++Na].
4.2.6.11. N-(4-((3-Methyl-1,4-dioxo-1,4-dihydronaphthalen-2-yl)selanyl)
phenyl)acetamide (4c). Compound 4c was prepared following
procedure II from 2-((4-aminophenyl)selanyl)-3-methylnaphthalene-
1,4-dione 2d (344 mg, 1 mmol) and acetic anhydride (2.0
equivalents). The progress of the product formation was followed by
TLC petroleum ether: EtOAc = 8:1, Rf = 0.43, purified by column silica
gel chromatography with petroleum ether: EtOAc = 8:3. Brown solid;
Yield: 288.75 mg (75%); mp 125–127 °C. 1H NMR (300 MHz, DMSO) δ
8.04–7.92 (m, 2H, Ar-H), 7.89–7.78 (m, 2H, Ar-H), 7.49 (qd, J = 6.7,
3.4 Hz, 4H, Ar-H), 2.07 (s, 3H, CH3), 2.03 (s, 3H, CH3); 13C NMR
(75 MHz, DMSO) δ 182.33, 181.60, 168.89, 149.50, 146.11, 139.51,
134.57, 134.01, 132.16, 126.96, 122.88, 120.29, 24.47, 17.96; MS
(ESI): m/z = found 384.83 [M+]; calcd. 385.02 [M+]; HRMS calcd. for
C19H15NO3Se [M++Na]: 408.01094, found 408.00933 [M++Na].
4.2.6.12. (E)-4-((4-((4-Bromobenzyl)selanyl)phenyl)amino)-4-oxobut-2-
enoic acid (5a). Compound 5a was prepared following procedure IV
from 4-((4-bromobenzyl)selanyl)aniline 2c (344 mg, 1 mmol) and
maleic anhydride (98 mg, 1 mmol). Its formation was monitored by
TLC chloroform: methanol = 8:1, Rf = 0.36, purified by column silica
gel chromatography with chloroform: methanol = 6:1. Yellow solid;
Yield: 412.66 mg (94%); mp 215–217 °C. 1H NMR (300 MHz, DMSO) δ
10.45 (s, 1H, COOH), 7.55 (d, J = 8.6 Hz, 2H, Ar-H), 7.46–7.37 (m, 4H,
Ar-H), 7.17 (d, J = 8.4 Hz, 2H, Ar-H), 6.45 (d, J = 12.1 Hz, 1H, =CH),
6.30 (d, J = 12.0 Hz, 1H, CH=), 4.04 (s, 2H, SeCH2); 13C NMR
(75 MHz, DMSO) δ 167.31, 163.71, 139.14, 138.42, 134.02, 132.06,
131.60, 131.35, 130.90, 124.14, 120.54, 120.15, 30.71; MS (ESI): m/
z = found 439.97 [M++1]; calcd. 438.93 [M+]; HRMS calcd. for
C17H14BrNO3Se [M++Na]: 461.92145, found 461.92022 [M++Na].
4.2.6.13. (Z)-4-((4-((3-Methyl-1,4-dioxo-1,4-dihydronaphthalen-2-yl)
selanyl)phenyl)amino)-4-oxobut-2-enoic acid (5b). Compound 5b was
prepared following procedure IV from 2-((4-aminophenyl)selanyl)-3-
methylnaphthalene-1,4-dione 2d (343 mg, 1 mmol) and maleic
anhydride (98 mg, 1 mmol). Its formation was monitored by TLC
chloroform: methanol = 8:1, Rf = 0.36, purified by column silica gel
chromatography with petrol chloroform: methanol = 6:1. Brown solid;
Yield: 401.31 mg (91%); mp 207–209 °C. 1H NMR (300 MHz, DMSO) δ
10.43 (s, 1H, COOH), 8.07–7.93 (m, 2H, Ar-H), 7.90–7.77 (m, 2H, Ar-
H), 7.63–7.51 (m, 2H, Ar-H), 7.29–7.10 (m, 2H, Ar-), 6.46 (dd,
J = 12.0, 2.6 Hz, 1H, =CH), 6.30 (dd, J = 12.0, 1.8 Hz, 1H, CH=),
2.30 (s, 1H, NH), 2.09 (s, 3H, CH3); 13C NMR (75 MHz, DMSO) δ
182.33, 181.52, 167.28, 149.94, 134.58, 134.35, 133.86, 132.27,
129.36, 128.67, 127.00, 126.78, 125.78, 124.20, 120.79, 18.10; MS
(ESI): m/z = found 464.03 [M++Na]; calcd. 441.01 [M+]; HRMS
calcd. for C21H15NO5Se [M++1]: 464.00077, found 463.99922
[M++Na].
4.2.6.14. 4-((4-((4-Bromobenzyl)selanyl)phenyl)amino)-4-oxobutanoic
acid (6a). Compound 62 was prepared following procedure IV from 2c
(344 mg, 1 mmol) and succinic anhydride (100 mg, 1 mmol). Its
formation was monitored by TLC chloroform: methanol = 8:1,
Rf = 0.36, purified by column silica gel chromatography with
chloroform: methanol = 6:1. White solid; Yield: 396.9 mg (90%); mp
258–260 °C. 1H NMR (300 MHz, DMSO) δ 12.13 (s, 1H, COOH), 10.00
(s, 1H, NH), 7.50 (d, J = 8.7 Hz, 2H, Ar-H), 7.45–7.33 (m, 4H, Ar-H),
7.17–7.12 (m, 2H, Ar-H), 4.14 (s, 2H, SeCH2), 2.54–2.51 (m, 4H, 2CH2);
13
C NMR (75 MHz, DMSO) δ 174.24, 170.64, 139.23, 134.26, 131.57,
131.32, 122.77, 120.09, 119.97, 31.53, 30.84, 29.20; MS (ESI): m/
z = found 463.99 [M++1]; calcd. 440.95 [M+]; HRMS calcd. for
C17H16BrNO3Se [M++Na]: 463.93710, found 463.93534 [M++Na].
S. Shaaban et al.
4.2.6.15. 4-((4-((3-Methyl-1,4-dioxo-1,4-dihydronaphthalen-2-yl)
selanyl)phenyl)amino)-4-oxobutanoic acid (6b). Compound 6b was
prepared following procedure IV from 2d (344 mg, 1 mmol)and
succinic anhydride (100 mg, 1 mmol). Its formation was monitored by
TLC chloroform: methanol = 8:1, Rf = 0.36, purified by column silica
gel chromatography with chloroform: methanol = 6:1. Brown solid;
Yield: 411.99 mg (93%); mp 208–210 °C. 1H NMR (300 MHz, DMSO) δ
12.11 (s, 1H, COOH), 10.04 (s, 1H, NH), 8.04–7.92 (m, 2H, Ar-H),
7.87–7.77 (m, 2H, Ar-H), 7.56–7.45 (m, 4H, Ar-H), 2.52 (dd, J = 6.8,
2.5 Hz, 4H, 2CH2), 2.07 (s, 3H, CH3); 13C NMR (75 MHz, DMSO) δ
182.33, 181.60, 173.27, 170.45, 149.63, 146.08, 139.38, 134.33,
134.04, 132.19, 126.97, 120.27, 51.82, 31.35, 28.87, 17.96; MS
(ESI): m/z = found 466.05 [M++Na]; calcd. 443.03 [M+]; HRMS
calcd. for C21H17NO5Se [M++Na]: 466.01642, found 466.01484
[M++Na].
4.2.6.16. (Z)-5-((4-((4-Bromobenzyl)selanyl)phenyl)imino)-1H-pyrrol-
2(5H)-one (9a). Compound 9a was prepared following procedure V
from 5a (439 mg, 1 mmol), acetic anhydride (3 ml) and 100 mg of
sodium acetate. The progress of the product formation was followed by
TLC petroleum ether: EtOAc = 8:1, Rf = 0.45, purified by column silica
gel chromatography with petroleum ether: EtOAc = 6:1. Yellow solid;
Yield: 324.17 mg (77%), mp 185–187 °C. 1H NMR (300 MHz, CDCl3) δ
7.47–7.29 (m, 4H, Ar-H), 7.25–7.14 (m, 2H, Ar-H), 7.04–6.96 (m, 2H,
Ar-H), 6.76 (d, J = 5.5 Hz, 1H, CH=), 6.62 (d, J = 5.5 Hz, 1H, CH=),
4.01 (s, 2H, SeCH2); 13C NMR (75 MHz, CDCl3) δ 166.94, 150.34,
143.16, 142.77, 137.50, 134.29, 134.02, 131.59, 130.49, 127.93,
125.94, 120.83, 31.52; MS (ESI): m/z = found 474.92 [M++CH3OH
+Na]; calcd. 420.92 [M+]; HRMS calcd. for C17H12BrNO2Se [M++1]:
475.93688, found 475.93544 [M++CH3OH+Na].
4.2.6.17. (E)-2-Methyl-3-((4-((5-oxofuran-2(5H)-ylidene)amino)phenyl)
selanyl)naphthalene-1,4-dione (9b). Compound 9b was prepared
following procedure V from 5b (441 mg, 1 mmol), acetic anhydride
(3 ml) and 100 mg of sodium acetate. The progress of the product
formation was followed by TLC petroleum ether: EtOAc = 8:1,
Rf = 0.46, purified by column silica gel chromatography with
petroleum ether: EtOAc = 6:1. Brown solid; Yield: 317.25 mg (75%);
mp 200–202 °C. 1H NMR (300 MHz, CDCl3) δ 8.09–7.94 (m, 2H, Ar-H),
7.69–7.59 (m, 2H, Ar-H), 7.57–7.42 (m, 2H, Ar-H), 7.35–7.20 (m, 2H,
Ar-H), 6.78 (d, J = 5.5 Hz, 1H, CH=), 6.67 (d, J = 5.5 Hz, 1H, CH=),
2.23 (s, 3H, CH3); 13C NMR (75 MHz, CDCl3) δ 182.44, 181.31, 169.11,
150.53, 146.11, 134.29, 133.89, 133.80, 133.66, 132.20, 132.00,
127.23, 126.81, 126.42, 121.59, 18.17; MS (ESI): m/z = found
478.03 [M++CH3OH+Na]; calcd. 423.00 [M+]; HRMS calcd. for
C21H13NO4Se [M+]: 420.92, found 477.9328 [M++CH3OH+Na].
4.2.6.18. 1-(4-((4-Bromobenzyl)selanyl)phenyl)pyrrolidine-2,5-dione
(10a). Compound 10a was prepared following procedure V from 6a
(441 mg, 1 mmol), acetic anhydride (3 ml) and 100 mg of sodium
acetate. The progress of the product formation was followed by TLC
petroleum ether: EtOAc = 8:1, Rf = 0.44, purified by column silica gel
chromatography with petroleum ether: EtOAc = 6:1. White solid; Yield:
321.48 mg (76%); mp 205–207 °C. 1H NMR (300 MHz, CDCl3) δ
7.49–7.41 (m, 2H, Ar-H), 7.34–7.26 (m, 2H, Ar-H), 7.15–7.08 (m, 2H,
Ar-H), 7.06–6.98 (m, 2H, Ar-H), 4.03 (s, 2H, SeCH2), 2.82 (s, 4H,
2CH2); 13C NMR (75 MHz, CDCl3) δ 174.87, 136.31, 132.99, 130.62,
130.17, 129.82, 129.51, 125.85, 119.87, 30.52, 27.39; MS (ESI): m/
z = found 445.93 [M++Na]; calcd. 422.94 [M+]; HRMS calcd. for
C17H14BrNO2Se [M++1]: 445.92653, found 445.92580 [M++Na].
4.2.6.19. 1-(4-((3-Methyl-1,4-dioxo-1,4-dihydronaphthalen-2-yl)selanyl)
phenyl)pyrrolidine-2,5-dione (10b). Compound 10b was prepared
following procedure V from 6b (443 mg, 1 mmol), acetic anhydride
(3 ml) and 100 mg of sodium acetate. The progress of the product
formation was followed by TLC petroleum ether: EtOAc = 8:1,
54
Bioorganic Chemistry 80 (2018) 43–56
Rf = 0.36, purified by column silica gel chromatography with
petroleum ether: EtOAc = 6:1. Brown solid; Yield: 323 mg (76%); mp
224–226 °C. 1H NMR (300 MHz, CDCl3) δ 8.07–7.95 (m, 2H, Ar-H),
7.68–7.59 (m, 2H, Ar-H), 7.57–7.51 (m, 2H, Ar-H), 7.21–7.15 (m, 2H,
Ar-H), 2.81 (s, 4H, 2CH2), 2.21 (s, 3H, CH3); 13C NMR (75 MHz, CDCl3)
δ 182.45, 181.23, 175.76, 150.72, 145.96, 133.76, 133.67, 132.18,
131.99, 131.50, 130.17, 127.25, 127.05, 126.80, 28.37, 18.24; MS
(ESI): m/z = found 447.38 [M++Na]; calcd. 425.02 [M+]; HRMS
calcd. for C21H15NO4Se [M++1]: 448.00585, found 448.00451
[M++Na].
4.2.6.20. (E)-Methyl 4-((4-((4-bromobenzyl)selanyl)phenyl)amino)-4-
oxobut-2-enoate (7a). Compound 7a was prepared following
procedure VI from 5a (439 mg, 1 mmol) in methanol (10 ml) and
conc.H2SO4 (200 µl). The progress of the product formation was
followed by TLC petroleum ether: EtOAc = 8:1, Rf = 0.43, purified by
column silica gel chromatography with petroleum ether: EtOAc = 6:1.
Brown solid; Yield: 389.58 mg (86%); mp 190–192 °C. 1H NMR
(300 MHz, DMSO) δ 11.05 (s, 1H, NH), 7.59 (d, J = 8.6 Hz, 2H, Ar-
H), 7.46–7.34 (m, 4H, Ar-H), 7.08–6.97 (m, 2H, Ar-H), 6.46 (d,
J = 13.4 Hz, 1H, CH=), 6.26 (d, J = 13.4 Hz, 1H, =CH), 4.02 (s, 2H,
SeCH2), 3.88 (s, 3H, OCH3); 13C NMR (75 MHz, DMSO) δ 162.60,
156.62, 135.86, 133.28, 132.95, 130.56, 126.72, 125.69, 120.22,
119.87, 115.87, 115.76, 48.12, 27.25; MS (ESI): m/z = found451.91
[M+]; calcd. 452.95 [M+]; HRMS calcd. for C18H16BrNO3Se [M++1]:
475.93710, found 475.93486 [M++Na].
4.2.6.21. (Z)-Methyl 4-((4-((3-methyl-1,4-dioxo-1,4-dihydronaphthalen-
2-yl)selanyl)phenyl)amino)-4-oxobut-2-enoate (7b). Compound 7b was
prepared following procedure VI from 5b (441 mg, 1 mmol) in
methanol (10 ml) and conc.H2SO4 (200 µl). The progress of the
product formation was followed by TLC petroleum ether:
EtOAc = 8:1, Rf = 0.36, purified by column silica gel
chromatography with petroleum ether: EtOAc = 6:1. Yellow solid;
Yield: 382.2 mg (84%); mp 208–210 °C. 1H NMR (300 MHz, CDCl3) δ
10.98 (s, 1H, NH), 8.05–7.92 (m, 2H, Ar-H), 7.66–7.57 (m, 2H, Ar-H),
7.54 (d, J = 8.7 Hz, 2H, Ar-H), 7.48–7.41 (m, 2H, Ar-H), 6.35 (dd,
J = 13.4, 3.3 Hz, 1H, CH=), 6.20–6.12 (dd, J = 13.4, 3.3 Hz, 1H,
=CH), 3.78 (s, 3H, OCH3), 2.12 (s, 3H, CH3); 13C NMR (75 MHz,
CDCl3) δ 181.41, 180.63, 166.33, 160.39, 148.36, 146.05, 139.42,
136.89, 133.68, 132.68, 132.49, 131.26, 131.04, 126.06, 125.73,
124.08, 123.46, 119.79, 51.87, 16.68; MS (ESI): m/z = found 478.04
[M++Na]; calcd. 455.03 [M+]; HRMS calcd. for C22H17NO5Se
[M++1]: 478.01642, found 478.01544 [M++Na].
4.2.6.22. Methyl 4-((4-((4-bromobenzyl)selanyl)phenyl)amino)-4-
oxobutanoate (8a). Compound 8a was prepared following
procedure VI from 4-((4-((4-bromobenzyl)selanyl)phenyl)amino)-4-
oxobutanoic acid 6a (441 mg, 1 mmol) in methanol (10 ml) and
conc.H2SO4 (200 µl). The progress of the product formation was
followed by TLC petroleum ether: EtOAc = 8:1, Rf = 0.41, purified
by column silica gel chromatography with petroleum ether:
EtOAc = 6:1. White solid; Yield: 395.85 mg (87%); mp 185–187 °C.
1
H NMR (300 MHz, CDCl3) δ 7.64 (s, 1H, NH), 7.37–7.20 (m, 6H, Ar-
H), 6.97–6.87 (m, 2H, Ar-H), 3.92 (s, 2H, SeCH2), 3.64 (s, 3H, OCH3),
2.73–2.53 (m, 4H, 2CH2); 13C NMR (75 MHz, CDCl3) δ 173.67,
169.75, 138.01, 137.74, 135.34, 131.45, 130.45, 124.10, 120.60,
120.17, 52.05, 32.16, 32.00, 29.19; MS (ESI): m/z = found 477.98
[M++Na]; calcd. 454.96 [M+]; HRMS calcd. for C18H18BrNO3Se
[M++1]: 477.95275, found 477.95070 [M++1].
Acknowledgements
This work was supported by the “Ministère des Affaires Etrangères:
the Embassy of France in Egypt-Institut Français en Egypte” and the
“Science and Technology Development Fund” by according to a
S. Shaaban et al.
postdoctoral fellowship to S. Shaaban. The authors thank the Conseil
Régional de Bourgogne (France) and the Ministère de l’Enseignement et
de la Recherche (France) for their financial supports. The authors thank
also the Egyptian Ministry of Higher Education and Mansoura
University for financial support. This article is based upon work from
COST Action CA16112-NutRedOx (Personalized Nutrition in aging society:
redox control of major age-related diseases), supported by COST
(European Cooperation in Science and Technology).
Appendix A. Supplementary material
Supplementary data associated with this article can be found, in the
online version, at https://doi.org/10.1016/j.bioorg.2018.05.019.
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