Synthesis of 2-amino-3-cyano-4H-pyran derivatives using GO-Fc@Fe3O4 nanohybrid as a novel recyclable heterogeneous nanocatalyst and preparation of tacrine-naphthopyran hybrids as AChE inhibitors

Article abstract of DOI:10.1007/s13738-020-02125-4

Abstract: GO-Fc@Fe₃O₄ nanohybrid as a powerful and reusable nanocatalyst was synthesized. The graphene oxide (GO) sheets with meta-chloroperoxybenzoic acid (mCPBA) were treatment, afterward were chemically modified with 4-Fc derivati

Full text of DOI:10.1007/s13738-020-02125-4

Journal of the Iranian Chemical Society (2021) 18:1455–1470
https://doi.org/10.1007/s13738-020-02125-4
ORIGINAL PAPER
Synthesis of 2‑amino‑3‑cyano‑4H‑pyran derivatives using GO‑Fc@
Fe3O4 nanohybrid as a novel recyclable heterogeneous nanocatalyst
and preparation of tacrine‑naphthopyran hybrids as AChE inhibitors
Sakineh Mozafarnia1,2 · Reza Teimuri‑Mofrad1 · Mohammad‑Reza Rashidi2
Received: 26 July 2020 / Accepted: 27 November 2020 / Published online: 3 January 2021
© Iranian Chemical Society 2021
Abstract
GO-Fc@Fe3O4 nanohybrid as a powerful and reusable nanocatalyst was synthesized. The graphene oxide (GO) sheets with
meta-chloroperoxybenzoic acid (mCPBA) were treatment, afterward were chemically modifed with 4-Fc derivative through
the ring-opening reaction between GO nanosheets and 4-ferrocenylbutylamine. Then, the fnal nanocatalyst was obtained
by synthesizing of Fe3O4 nanoparticles onto the modifed GO surface. The structure and morphology of GO-Fc@Fe3O4
nanohybrid were characterized using diferent analysis, such as FT-IR, XRD, FE-SEM, EDX, and VSM techniques. Then,
2-amino-3-cyano-4H-pyran derivatives (4a–l) as a synthetic precursor were synthesized via multi-component reaction in the
presence of GO-Fc@Fe3O4 nanohybrid as a novel heterogeneous nanocatalyst. Finally, according to the importance of fnding
a solution to treat Alzheimer’s disease, 14-aryl-10,11,12,14-tetrahydro-9H-benzo[5,6]chromeno[2,3-b]-quinolin-13-amines
(5a–l) as new tacrine-naphthopyran hybrid analogs were designed and prepared as acetylcholinesterase inhibitors. These
compounds were synthesized via Friedländer reaction of 2-amino-3-cyano-4H-pyran derivatives (4a–l) with cyclohexanone.
HAChE inhibition assay was carried out in vitro on the synthesized compounds 5a–l. Among them, compound 5f exhibited
potent hAChE inhibitors with IC50 values of 0.16 µM. Also, the molecular docking and kinetic studies performed for a better
understanding of compound 5f as a representative compound.
Graphic abstract
Keywords GO-Fc@Fe3O4 nanohybrid · Multi-component reaction · Solvent-free · Tacrine hybrid · Molecular modeling
* Reza Teimuri-Mofrad
teymouri@tabrizu.ac.ir
Extended author information available on the last page of the article
Vol.:(0123456789)
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
Abbreviations
a variety of biological activities as anti-hypertensive, anti-
coagulant, anti-allergic, anticancer, and antiviral activities.
Also, organic photochromic naphthopyrans have an indus-
trial applications of technology and used in manufacturing
in plastic photochromic lenses (T-type photochromism).
Due to the medicinal properties of pyran derivatives, it is
important to use appropriate synthesis methods. One of the
methods used is synthesis via a multi-component reaction
in the presence of a suitable catalyst [37–45]. On the other
hand, multi-component, solvent-free, and reusable heteroge-
neously catalyzed reactions are factors for the principles of
green chemistry. Multi-component reactions are very strong
and efective bond-forming tools in organic chemistry with
high atom efciency, time and energy saving, friendly envi-
ronment, and rapid and simple efciency. Also, the catalyst
plays a signifcant role in selectivity and determining yield.
So, the creation of a reusable, mild, inexpensive catalyst for
multi-component reactions has attracted interest. The cata-
lysts have been used for multi-component reactions, includ-
ing ZnAl2O4–Bi2O3, SiO2 NPs, Fe3O4@SiO2@imidazol-
bisFc[HCO3], Fe3O4@SiO2-BenzIm-Fc[Cl]/NiCl2, SiO2@
Imid-Cl@Fc, and silica-bonded aminoethylpiperazine [33,
46–51]. The use of an efcient and stable heterogeneous
catalyst as reusable and environmentally friendly substances
has a very important role in organic synthesis. Also, the
immobilization of catalyst on the solid supports have advan-
tages such as non-toxicity, low solubility, increased reactions
selectivity, and easy handling and led to the widespread use
of these catalysts. Catalysts immobilized on the nanoparticle
supports are more attractive, because of their higher selectiv-
ity and activity. Magnetite (Fe3O4) nanoparticle with unique
properties such as high surface area, low toxicity, superpara-
magnetism, thermal stability, low cost, easy separation from
the reaction mixture and recyclability used in the design and
synthesis of reusable heterogeneous catalysts [49, 52–56].
Ferrocene derivatives have become very important for
their applications such as redox electrochemistry, catalysis,
biology, materials science, and nonlinear optical materi-
als. Also, characteristics such as thermal stability, low cost,
high tolerance to moisture, and oxygen make these materials
attractive [57, 58]. Graphene oxide (GO) is one of the most
important derivatives and the oxidized forms of graphene.
GO has diferent oxygen groups such as hydroxyl, carboxyl,
epoxide and carbonyl on its surface. Also, it has proper-
ties such as good chemical and mechanical stability, high
conductivity and a special surface, and various applications
such as energy storage materials, polymer nanocomposites
and electrocatalysis [59–61].
AD
Alzheimer’s disease
ACh
AChE
Acetylcholine
Acetylcholinesterase
BuChE Butyrylcholinesterase
ChEI
PAS
CAS
GO
Cholinesterase inhibitor
Peripheral anionic site
Catalytic active site
Graphene oxide
mCPBA meta-Chloroperoxybenzoic acid
Introduction
Alzheimer’s disease (AD) is an irreversible and multifacto-
rial neurodegenerative illness and the main cause of demen-
tia in elderly people [1–3]. AD begins with memory loss
in the early stages and gradually develops in the advanced
stages of cognitive impairment, causing severe behavioral
abnormalities and eventually leading to death [4–7]. Due
to the increased life expectancy and the number of elderly
people, AD has become an important public health issue [8,
9]. AD pathophysiology is not yet fully clear, but factors
other than age and heredity are involved as a shortage of ace-
tylcholine (ACh), beta-amyloid (Aβ) aggregates, τ-protein
aggregation, oxidative stress, mitochondrial dysfunction, and
neuroinfammation [10–14]. There is not a defnitive cure for
AD, but one efective therapy for AD is increasing ACh
levels in the brain by the inhibition of acetylcholinesterase
(AChE) [15–17]. Acetylcholinesterase inhibitors including
donepezil, rivastigmine, tacrine, and galantamine were used
to treat AD [18, 19]. Tacrine was the frst developed revers-
ible cholinesterase inhibitor (ChEI) that was approved by the
US FDA for the therapy for AD in 1993. Nevertheless, due
to hepatotoxicity and peripheral side efects, it was approxi-
mately removed from the market. Tacrine is synthesized by
Friedländer-type cyclocondensation of o-aminobenzonitrile
with cyclohexanone [20–25]. Currently, research is ongoing
on tacrine to design and synthesis new tacrine-based drugs
via Friedländer reaction for modifcation on the structure
of tacrine by the exchange of benzene rings in tacrine with
other synthetic compounds [7, 26, 27]. The Friedländer
reaction has been known for more than a century and is a
well-known and simplest method for synthesis of diferent
nitrogen-containing heterocyclic compounds such as quino-
line derivatives, tacrine analogs, acridines, pyridines, camp-
tothecins, and phenanthrolines [28–30].
Pyran is one of the main core units in several natural
and synthetic products. Organic molecules with 4H-pyran
ring have biological and potential medical such as anti-
allergic, antitumor, antibacterial, anti-HIV, antifungal, anti-
coagulant, anti-cancer, anti-infammatory, and neurodegen-
erative disorders [31–36]. Naphthopyran derivatives show
In this work, GO-Fc@Fe3O4 nanohybrid as a new, power-
ful, and reusable nanocatalyst was synthesized. The modi-
fed GO nanosheets provide a large available surface area
with several acidic and basic sites including carboxylic acid,
hydroxyl, Fc rings on the GO-Fc surface. This nanocatalyst
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was used to synthesize of 2-amino-3-cyano-4H-pyran
derivatives (4a–l) under solvent-free conditions (Fig. 1).
The desired products were obtained with good yield and
in a short time. Compounds 4a–l were used as precursors
in the Friedländer reaction with cyclohexanone and under
the usual experimental conditions were obtained 14-aryl-
10,11,12,14-tetrahydro-9H-benzo[5,6]chromeno[2,3-b]-qui-
nolin-13-amines derivatives (5a–l) as new tacrine analogs
for inhibition of acetylcholinesterase enzyme (Fig. 1). The
amount of hAChE and hBuChE enzymes inhibitory activ-
ity was investigated for these compounds and the results
showed that this series of compounds have good inhibition
only against hAChE enzyme.
progress was controlled by TLC and detected by UV light
(254 nm). 1H NMR and 13C NMR spectra were registered on
Bruker Spectrospin Avance 400 and 100 MHz spectrometers
in DMSO-d6 solvent, respectively. All chemical shifts were
reported as δ (ppm), and coupling constants (J) were given in
Hz. The Bruker Tensor 27 tool was used to record the FT-IR
spectra of the synthesized compound on the KBr pellets and
expressed in cm−1. Using an electric apparatus MEL-TEMP
model 1202 was measured the melting point. Elemen-tar
Vario EL III tool was used for Elemental analysis (C, H,
N). FESEM (TESCAN MIRA3), EDX spectroscopy (TES-
CAN MIRA3), and XRD analysis (PANalytica X pertPRO
(Germany) instrument with Cu-Ka radiation (0.15406 nm) at
accelerating voltage of 45 kV) were used to study the mor-
phology and structure of the nanoparticles. Magnetization
measurement was performed with a model 155 alternative
gradient force magnetometer at room temperature.
Experimental
Materials and instruments
Preparation of GO
Commercially available compounds were bought from
Merck and Sigma-Aldrich. Commercial compounds contain:
graphene oxide (GO), sodium nitrate (NaNO3) (CAS num-
ber: 7631-99-4), potassium permanganate (KMnO4) (CAS
number: 7722-64-7), sulfuric acid (H2SO4) (CAS number:
7664-93-9), hydrogen peroxide solution (H2O2) (CAS num-
ber: 7722-84-1), meta-chloroperoxybenzoic acid (mCPBA)
(CAS number: 937-14-4), glacial acetic acid (CAS num-
ber: 64-19-7), iron(II) chloride tetrahydrate (FeCl2.4H2O)
(CAS number: 13478-10-9), iron(III) chloride hexahydrate
(FeCl3.6H2O) (CAS number: 10025-77-1), 2-naphthol (CAS
number: 135-19-3), 6-bromo-2-naphthol (CAS number:
15231-91-1), malononitrile (CAS number: 109-77-3), ben-
zaldehyde (CAS number: 100-52-7), 4-nitrobenzaldehyde
(CAS number: 555-16-8), 4-methylbenzaldehyde (CAS
number: 104-87-0), 4-isopropylbenzaldehyde (CAS number:
122-03-2), 4-chlorobenzaldehyde (CAS number: 104-88-1),
4-bromobenzaldehyde (CAS number: 1122-91-4), 2-chlo-
robenzaldehyde (CAS number: 89-98-5), 3-bromobenzal-
dehyde (CAS number: 3132-99-8), 4-fuorobenzaldehyde
(CAS number: 459-57-4), thiophene-2-carbaldehyde (CAS
number: 98-03-3), cyclohexanone (CAS number: 108-94-
1), Aluminum chloride (AlCl3) (CAS number: 7446-70-0),
1,2-dichloroethane (CAS number: 107-06-2). The reaction
Hummer’s method was used to prepare GO nanosheets by
oxidizing graphite fakes [62]. In brief, a round bottom fask
was charged with graphite fake (2.5 g) and NaNO3 (1.25 g).
The fask was then immersed in an ice bath and 65 ml of
concentrated H2SO4 was added to a fask and stirred for 1 h.
Then, fnely ground KMnO4 was added in small portions
into the above mixture under vigorous stirring at 0 °C. Vig-
orous stirring was continued for a further 72 h at 25 °C.
After adding 175 ml of distilled water, 100 ml of HCl (10%)
and 30 ml of H2O2 (30%) were slowly added to the above
mixture and stirred for 2 h. After that, the mixture was cen-
trifuged and washed with a large amount of deionized water
to give GO powders.
Preparation of over‑oxidized GO (GO‑Epo)
An amount of 150 mg of mCPBA was added to a suspension
of GO in DI water (25 ml, 3 mg/ml) and stirred at ambient
temperature for 24 h. After this time, the reaction mixture
was washed with dichloromethane to remove all the unre-
acted mCPBA. Then, the aqueous phase was centrifuged to
give the over-oxidized GO nanosheets (GO-Epo) [63].
Fig. 1 Design and synthesis of 2‐amino‐3‐cyano‐4H‐pyran (4a–l) and 14-aryl-10,11,12,14-tetrahydro-9H-benzo[5,6]chromeno[2,3-b]-quinolin-
13-amines (5a–l) derivatives
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and washed with EtOH. The synthesized compounds were
obtained as solids.
Functionalization of GO‑Epo with 4‑ferrocenylbutyl‑
amine (GO‑Fc)
An amount of 0.3 g of 4-ferrocenylbutylamine [64] was dis-
solved in 35 ml of ethanol and the resulted solution was then
added to a suspension of GO-Epo in water (0.1 g in 30 ml
water). The resulting mixture was refuxed for 72 h [60].
At the end of the reaction, the mixture was centrifuged and
washed with water and dichloromethane, respectively. After
vacuum drying, the GO-Fc was obtained as a black powder.
14‑(Phenyl)‑10,11,12,14‑tetrahydro‑9H‑benzo[5,6]
chromeno[2,3‑b]‑quinolin‑13‑amine (5a)
Yield 92%; white solid; m.p.>250 °C (mp.>260 °C [22]).
14‑(4‑Nitrophenyl)‑10,11,12,14‑tetrahydro‑9H‑benzo[5,6]
chromeno[2,3‑b]‑quinolin‑13‑amine (5b)
Synthesis of GO‑Fc@Fe3O4 nanohybrid
Yield 73%; white solid; m.p. > 250 °C (mp. 258–260 °C
[66]).
A round bottom two necked flask was charged with
FeCl3·6H2O (1.76 g, 6.5 mmol) and FeCl2·4H2O (0.65 g,
3.25 mmol). A mixture of glacial acetic acid (1.5 ml) and
water (50 ml) was added to the above mixture under vig-
orous stirring. The appropriate amount of GO-Fc suspen-
sion was then added to the above fask and the temperature
was raised from 25 to 80 °C. After the temperature reached
to 80 °C, 10 ml of ammonia was added into the fask and
allowed to stir for an additional 15 min at 80 °C. Finally,
GO-Fc@Fe3O4 nanohybrid was separated by an exter-
nal magnetic feld and was washed with distilled water and
methanol, respectively [65].
14‑(4‑Methylphenyl)‑10,11,12,14‑tetrahydro‑9H‑benzo[5,6]
chromeno[2,3‑b]‑quinolin‑13‑amine (5c)
Yield 59%; white solid; m.p. 223–227 °C (mp. 224–226 °C
[66]).
14‑(4‑Isopropylphenyl)‑10,11,12,14‑tetrahy‑
dro‑9H‑benzo[5,6]chromeno[2,3‑b]‑quinolin‑13‑amine (5d)
Yield 57%; white solid; m.p. 239–244 °C. FTIR (KBr) ν
3492, 3355, 2931, 1617, 1444, 1227 cm−1; 1H NMR: 1.04
(d, 3H, 3J=0.9 Hz, CH3), 1.05 (d, 3H, 3J=0.9 Hz,, CH3),
1.71 (br, 4H, CH2), 2.21–2.25 (m, 1H, CH2), 2.38–2.42
(m, 1H, CH2), 2.63–2.64 (m, 2H, CH2), 2.65–2.70 (m ,1H,
CH), 6.17 (s, 1H, CH), 6.70 (br, 2H, NH2), 7.04 (d, 2H,
3J = 8.2 Hz, Ar–H), 7.42–7.47 (m, 4H, Ar–H), 7.55–7.59
(m, 1H, Ar–H), 7.91 (d, 2H, 3J=9.1 Hz, Ar–H), 8.36 (d, 1H,
3J=8.5 Hz, Ar–H); 13C NMR: 21.5, 21.6, 22.7, 23.6, 29.8,
32.8, 34.0, 99.7, 112.5, 117.3, 118.0, 123.4, 124.7, 126.2,
126.9, 127.5, 128.5, 129.0, 130.5, 130.6, 141.4, 146.5,
147.8, 149.1, 153.0, 153.3; Anal. Calcd. for C29H28N2O
(420.55): C 82.82, H 6.71, N 6.66; Found: C 82.89, H 6.70,
N 6.59%.
General procedure for the preparation
of 2‑amino‑3‑cyano‑4H‑pyran derivatives (4a–l)
A reaction vial was charged with 2-naphthol (1a) or
6-bromo-2-naphthol (1b) (1 mmol), aromatic aldehyde
(2a–j) (1 mmol), malononitrile (3) (1.2 mmol), and GO-Fc@
Fe3O4 (10 mg) as magnetic nanocatalyst. The mixture was
heated at 100 °C under solvent-free conditions. After com-
pletion of the reaction (monitored by TLC), an appropriate
amount of ethanol was added and the mixture heated for
5 min. Then, the magnetic nanocatalyst was separated by
applying an external magnet. The crude solid was collected
by fltration and then recrystallized from ethanol to give the
pure compound.
14‑(4‑Chlorophenyl)‑10,11,12,14‑tetrahydro‑9H‑benzo[5,6]
chromeno[2,3‑b]‑quinolin‑13‑amine (5e)
General procedure for the synthesis
of 14‑aryl‑10,11,12,14‑tetrahydro‑9H‑benzo[5,6]
chromeno[2,3‑b]‑quinolin‑13‑amines derivatives
(5a–l)
Yield 65%; white solid; m.p.>250 °C. FTIR (KBr) ν 3418,
1
3206, 2940, 1648, 1478, 1226 cm−1; H NMR: 1.69–1.72
(m, 4H, CH2), 2.21–2.25 (m,1H, CH2), 2.40–2.45 (m,1H,
CH2), 2.72–2.73 (m, 2H, CH2), 6.53 (s, 1H, CH), 7.26 (d,
2H, 3J=8.5 Hz, Ar–H), 7.44 (t,1H, 3J=7.8 Hz, Ar–H), 7.49
(d,1H, 3J=8.9 Hz, Ar–H), 7.55 (t, 1H, 3J=7.8 Hz, Ar–H),
7.63 (d, 2H, 3J=8.4 Hz, Ar–H), 7.92–7.99 (m, 2H, Ar–H),
AlCl3 (1.5 equiv) and dry 1,2-dichloroethane (10 ml) were
mixed at ambient temperature under argon atmosphere for
30 min. Then, compounds 4a–l (1 equiv) and cyclohex-
anone (1.5 equiv) were added to them and hated at refux
for 24 h. Then, using a rotary evaporator was evaporated the
solvent and NaOH 10% was added to the residual sediment
and stirred for 30 min. The resulted mixture was fltered
3
8.36 (d, 1H, J = 8.5 Hz, Ar–H); 13C NMR: 22.1, 22.3,
23.0, 32.0, 34.4, 99.0, 112.3, 115.8, 116.5, 117.8, 119.3,
124.3, 126.1, 128.7, 129.3, 129.6, 129.9, 130.5, 131.6,
143.0, 148.0, 148.8, 151.2, 152.3, 154.8. Anal. Calcd. for
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
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C26H21ClN2O (412.91): C 75.63, H 5.13, N 6.78; Found: C
75.52, H 5.09, N 6.74%.
14‑(3‑Bromophenyl)‑10,11,12,14‑tetrahydro‑9H‑benzo[5,6]
chromeno[2,3‑b]‑quinolin‑13‑amine (5i)
14‑(2‑Cholorophenyl)‑10,11,12,14‑tetrahy‑
dro‑9H‑benzo[5,6]chromeno[2,3‑b]‑quinolin‑13‑amine (5f)
Yield 72%; white solid; m.p. > 250 °C. FTIR (KBr) ν
3482, 3348, 2933, 1624, 1439, 1231 cm−1; 1H NMR: 1.71
(br,4H, CH2), 2.21–2.25 (m,1H, CH2), 2.37–2.41(m, 1H,
CH2), 2.53–2.58 (m, 2H, CH2), 6.08 (s, 2H, NH2), 6.11 (s,
1H, CH), 7.12–7.16 (t, 1H, 3J = 7.8 Hz, Ar–H), 7.24–7.26
(m, 1H, Ar–H), 7.41–7.45 (m, 3H, Ar–H), 7.57–7.61(m,
1H, Ar–H), 7.75 (br, 1H, Ar–H), 7.89–7.92 (m, 2H,
Ar–H), 8.29 (d, 1H, 3J = 8.5 Hz, Ar–H); 13C NMR: 22.5,
22.7, 23.5, 32.3, 35.4, 99.4, 113.2, 117.6, 118.4, 122.2,
123.3, 125.2, 127.1, 127.7, 129.3, 129.9, 130.0, 130.5,
130.9, 131.0, 131.2, 147.6, 149.3, 152.0, 153.2, 155.3;
Anal. Calcd. for C26H21BrN2O (457.36): C 68.28, H 4.63,
N 6.12; Found: C 68.31, H 4.57, N 6.16%.
Yield 92%; white solid; m.p.>250 °C. FTIR (KBr) ν 3466,
3332, 2926, 1625, 1445, 1228 cm−1; 1H NMR: 1.70 (S, 4H,
CH2), 2.20–2.24 (m, 1H, CH2), 2.32–2.39 (m, 1H, CH2),
2.53–2.58 (m, 2H, CH2), 5.66-5.67 (br, 2H, NH2), 6.16
(s, 1H, CH), 7.10–7.14 (m,1H, Ar–H), 7.16–7.20 (m, 1H,
Ar–H), 7.35 (dd, 1H, 3J=7.8, 1.2 Hz, Ar–H), 7.39–7.46 (m,
3H, Ar–H), 7.52–7.55 (m, 1H, Ar–H), 7.89–7.92 (m, 2H,
3
Ar–H), 8.25 (d, 1H, J = 8.0 Hz, Ar–H); 13C NMR: 22.0,
22.2, 22.9, 31.9, 34.1, 98.2, 112.4, 116.0, 117.9, 122.7,
124.3, 126.9, 127.0, 128.5, 128.6, 129.4, 129.5, 130.2,
130.4, 130.8, 131.5, 141.6, 149.0, 151.3, 152.7, 154.5; Anal.
Calcd. for C26H21ClN2O (412.91): C 75.63, H 5.13, N 6.78;
Found: C 75.70, H, 5.15, N 6.81%.
3‑Bromo‑14‑(4‑fuorophenyl)‑10,11,12,14‑tetrahy‑
dro‑9H‑benzo[5,6]chromeno [2,3‑b]‑quinolin‑13‑amine (5j)
14‑(4‑Bromophenyl)‑10,11,12,14‑tetrahydro‑9H‑benzo[5,6]
chromeno[2,3‑b]‑quinolin‑13‑amine (5 g)
Yield 87%; white solid; m.p. > 250 °C. FTIR (KBr) ν
3480, 3381, 2929, 1618, 1442, 1233 cm−1; 1H NMR: 1.70
(br, 4H, CH2), 2.19–2.23 (m, 1H, CH2), 2.32-2.37 (m,
1H, CH2), 2.57 (br, 2H, CH2), 6.04 (s, 2H, NH2), 6.12 (s,
Yield 81%; white solid; m.p.>250 °C. FTIR (KBr) ν 3453,
3406, 2928, 1629, 1442, 1231 cm−1; 1H NMR: 1.70 (br, 4H,
CH2), 2.19–2.23 (m, 1H, CH2), 2.37–2.42 (m, 1H, CH2),
2.571–2.578 (m, 2H, CH2), 6.05 (s, 2H, NH2), 6.13 (s, 1H,
3
1H, CH), 7.00 (t, 2H, J = 8.8 Hz, Ar–H), 7.46–7.50 (m,
3
3H, Ar–H), 7.70 (dd, 1H, J = 6.9, 2.0 Hz, Ar–H), 7.88
3
3
3
CH), 7.35–7.46 (m, 6H, Ar–H), 7.55 (t, 1H, J = 7.2 Hz,
(d, 1H, J = 9 Hz, Ar–H), 8.19 (d, 1H, J = 2 Hz, Ar–H),
Ar–H), 7.87–7.90 (m, 2H, Ar–H), 8.29 (d, 1H, 3J=8.5 Hz,
Ar–H); 13C NMR: 22.1, 22.2, 23.0, 32.0, 34.5, 98.8, 112.3,
117.3, 117.8, 119.3, 123.0, 124.3, 126.8, 128.5, 128.9,
129.7, 130.2, 130.6, 131.0, 144.1, 148.8, 151.2, 152.3,
154.8; Anal. Calcd. for C26H21BrN2O (457.36): C 68.28, H
4.63, N 6.12; Found: C 68.22, H 4.61, N 6.15%.
8.24 (d, 1H, J = 9.1 Hz, Ar–H); 13C NMR: 22.1, 22.3,
3
23.1, 32.0, 34.2, 99.8, 111.4, 113.5, 114.6, 115.3, 116.8,
122.0, 124.3, 127.4, 129.0, 129.8, 130.2, 132.6, 131.1,
131.6, 140.5, 148.8, 151.4, 152.2, 154.7. Anal. Calcd. for
C26H20BrFN2O (475.35): C 65.69, H 4.24, N 5.89; Found:
C 65.54, H 4.23, N 5.83%.
3‑Bromo‑14‑(3‑bromophenyl)‑10,11,12,14‑tetrahy‑
dro‑9H‑benzo[5,6]chromeno [2,3‑b]‑quinolin‑13‑amine
(5h)
14‑(4‑Fluorophenyl)‑10,11,12,14‑tetrahydro‑9H‑benzo[5,6]
chromeno[2,3‑b]‑quinolin‑13‑amine (5k)
Yield 78%; white solid; m.p.>250 °C. FTIR (KBr) ν 3425,
Yield 50%; white solid; m.p. > 250 °C. FTIR (KBr) ν
3488, 3354, 2926, 1617, 1439, 1225 cm−1; 1H NMR: 1.69
(br, 4H, CH2), 2.20-2.24 (m, 1H, CH2), 2.38-2.42 (m,
1H, CH2), 2.57 (br, 2H, CH2), 6.05 (s, 2H, NH2), 6.13 (s,
1
3152, 2934, 1651, 1412, 1230 cm−1; H NMR: 1.73-1.74
(m, 4H, CH2), 2.25 (br, 1H, CH2), 2.40 (br, 1H, CH2), 2.73
3
(br, 2H, CH2), 6.39 (s, 1H, CH), 7.19 (t, 1H, J = 7.8 Hz,
Ar–H), 7.34 (dd, 1H, 3J=7.1, 0.7 Hz, Ar–H), 7.44 (d, 1H,
3J=7.8 Hz, Ar–H), 7.61 (d, 1H, 3J=9.0 Hz, Ar–H), 7.82
(dd, 2H, 3J=7.0, 1.9 Hz, Ar–H), 7.89 (s, 1H, Ar–H), 8.02
(d, 1H, 3J=9.1 Hz, Ar–H), 8.23 (d, 1H, 3J=9.1 Hz, Ar–H),
8.30 (d, 1H, 3J=1.0 Hz, Ar–H);13C NMR: 22.5, 22.7, 23.5,
32.4, 35.4, 99.3, 111.4, 113.5, 117.6, 121.8, 122.2, 124.3,
126.2, 128.4, 128.9, 130.2, 130.5, 130.9, 131.4, 131.9,
132.3, 147,6, 149.3, 152.1, 153.1, 155.4. Anal. Calcd. for
C26H20 Br2N2O (536.26): C 58.23, H 3.76, N 5.22; Found:
C 58.10, H 3.72, N 5.18%.
1H, CH), 7.00 (t, 2H, J = 8.7 Hz, Ar–H), 7.38–7.44 (m,
3
2H, Ar–H), 7.51–7.57 (m, 3H, Ar–H), 7.86-7.90 (m, 2H,
Ar–H), 8.32 (d, 1H, 3J = 8.5 Hz, Ar–H); 13C NMR: 22.1,
22.3, 23.0, 32.0, 34.2, 99.2, 112.3, 114.8, 115.0, 117.80,
117.86, 123.0, 124.3, 126.7, 128.5, 128.8, 129.2, 129.3,
130.2, 130.6, 140.9, 148.8, 151.1, 152.2, 154.8; Anal.
Calcd. for C26H21FN2O (396.46): C 78.77, H 5.34, N 7.06;
Found: C 78.84, H, 5.37, N 7.07%.
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
14‑(Thiophen‑2‑yl)‑10,11,12,14‑tetrahydro‑9H‑benzo[5,6]
chromeno[2,3‑b]‑quinolin‑13‑amine (5l)
Kinetic assay
Using Ellman’s method, kinetic studies of inhibition on
hAChE were performed to obtain the inhibition model and
value Ki. In this study, the enzyme relative speed on various
concentrations of the substrates (0.1–1 mM) and two various
concentrations of inhibitor 5f (5×10−7–1×10−6 mM) was
determined. Linear regression was used for the calculation
of Lineweaver–Burk plots, and all Data analysis was done
using Microsoft Excel 2013 [69].
Yield 52%; white solid; m.p.>250 °C. FTIR (KBr) ν 3464,
3356, 2932, 1622, 1444, 1235 cm−1; 1H NMR: 1.72 (br, 4H,
CH2), 2.30–2.32 (m, 1H, CH2), 2.39–2.43 (m, 1H, CH2),
2.53–2.58 (m, 2H, CH2), 6.15 (s, 2H, NH2), 6.50 (s, 1H,
CH), 6.76–6.78 (m, 1H, Ar–H), 7.12–7.15 (m, 2H, Ar–H),
3
3
7.39 (d, 1H, J = 8.8 Hz, Ar–H), 7.44 (t, 1H, J = 7.2 Hz,
Ar–H), 7.57–7.61 (m, 1H, Ar–H), 7.88–7.93 (m, 2H, Ar–H),
3
8.40 (d, 1H, J = 8.4 Hz, Ar–H); 13C NMR: 22.1, 22.32,
23.0, 30.6, 32.0, 98.8, 112.2, 117.5, 117.8, 124.2, 125.9,
126.7, 128.4, 128.9, 130.2, 130.7, 148.4, 148.7, 151.3,
152.3, 154.9; Anal. Calcd. for C24H20N2OS (384.50): C
74.97, H 5.24, N 7.29 S 8.34; Found: C 74.87, H, 5.21, N
7.24 S 8.29%.
Docking studies
HAChE Structure (pdb ID: 4Ey7) was taken of the Protein
Data Bank [70, 71]. To set up and perform blind docking
calculations between drug and hAChE were used AutoGrid
4 and AutoDock 4 [72, 73]. At frst, water molecules were
deleted. Lamarckian genetic algorithms were applied to
carry out docking calculations as implemented in AutoDock.
The results from AutoDock was rendered with Discovery
studio 4.5.
hAChE and hBuChE inhibition assay
Using Ellman’s method [67] was measured inhibitory
capacity of new compounds on ChE activity. Fresh blood
was used to make the enzymes used in this procedure [68].
Synthetic compounds were solved in DMSO and they were
then diluted with bufer (0.1 M, pH=8) to reach fnal con-
centrations. Phosphate bufer (0.1 M, pH 8, 550 µl), DTNB
(3.5 mM, 150 µl), the substrate (acetylthiocholine or butyl-
thiocholine) (7 mM, 150 µl) and diferent concentrations
of inhibitors (150 µl) and phosphate bufer (0.1 M, 700 µl),
DTNB (3.5 mM, 150 µl) and diferent concentrations of
inhibitors (150 µl) were dumped in test cuvettes and con-
trol cuvettes, respectively. After 5 min, it reached to 37 °C,
enzyme (50 µl) was added. All experiments were performed
by spectrophotometer Helios-Zeta (Thermospectronic, Cam-
bridge, U.K.) at 412 nm in 5 min.
Results and discussion
Synthesis and characterization of GO‑Fc@Fe3O4
nanohybrid
GO-Fc@Fe3O4 nanohybrid as a powerful and reusable nano-
catalyst for the synthesis of 2-amino-4H-pyran derivatives
was synthesized according to scheme 1. Firstly, over oxi-
dized graphene oxide (GO-Epo) was prepared by the treat-
ment of GO with mCPBA in an aqueous medium at room
temperature [63]. This step provides a much reactive site
Scheme 1 Synthesized process of GO-Fc@Fe3O4 nanohybrid
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
1461
for surface modifcation of GO nanosheets. Afterward, GO-
Epo nanosheets were chemically modifed with Fc derivative
through the ring-opening reaction between GO nanosheets
and 4-ferrocenylbutylamine as a nucleophile [60]. Then,
Fe3O4 nanoparticles were synthesized onto the modifed
GO surface via a simple co-precipitation of Fe2+ and Fe3+
under an alkaline solution at 80 °C [65]. The fnal magnetic
nanocatalyst denoted as GO-Fc@Fe3O4.
FT-IR spectra of GO, GO-Fc, and GO-Fc@Fe3O4
nanohybrid are depicted in Fig. 2. As shown in Fig. 2,
GO nanosheets show several characteristic peaks at about
3400 cm−1, 2900 cm−1, 2800 cm−1, 1700 cm−1,1600 cm−1
which are related to the OH, aromatic C–H bond, aliphatic
C–H bond, C=O, and C=C bonds, respectively [74]. As
indicated in this fgure, in the FT-IR spectrum of GO-Fc, the
appearance of a new peak at about 480 cm−1 can be assigned
to the Fe-Cp bond in Fc rings. This new peak proved the suc-
cess of the grafting of Fc groups onto GO nanosheets. Also,
the FT-IR spectrum of GO-Fc@Fe3O4 nanohybrid exhibits
a strong IR peak at around 570 cm−1 which is related to the
stronger bending vibration of Fe–O in Fe3O4 nanoparticles
[75]. These fndings indicate that the Fe3O4 nanoparticles
were successfully synthesized by the in situ nanoparticle
growth method onto the GO-Fc surface.
Fig. 3 XRD patterns of GO, GO-Fc, GO-Fc@Fe3O4 nanohybrid
The XRD pattern of GO-Fc@Fe3O4 nanohybrid con-
firmed the presence of Fe3O4 nanoparticles and GO-Fc
nanosheets related difraction peaks. Broad XRD peaks of
GO-Fc@Fe3O4 nanohybrid compared to GO-Fc nanosheets
can be attributed to the growth of Fe3O4 nanoparticles
between GO layers.
The XRD patterns of GO, GO-Fc, GO-Fc@Fe3O4 nano-
hybrid are shown in Fig. 3. The XRD pattern of GO exhibits
a sharp difraction peak at 2θ = 11.5° which is attributed
to the (001) peak (d~0.77 nm). After modifcation of GO
surface with Fc moieties, the XRD peak position and shape
of GO-Fc changes compared to the XRD peak of GO. These
changes may be due to the presence of Fc rings onto the
GO surface which disrupts the crystalline structure of GO
nanosheets.
FE-SEM micrographs of GO, GO-Fc, GO-Fc@Fe3O4
nanohybrid are depicted in Fig. 4 that tangible changes are
observed in the FE-SEM micrographs of them. The FE-SEM
image of GO (Fig. 4a) exhibits a layered structure with wrin-
kles visible. The wrinkled structure of GO can be due to the
presence of oxygenic functional groups on its surface. After
modifcation of GO surface with Fc moieties, the GO-Fc
nanosheets show rich-wrinkle morphology and also are
increased surface turbidity that is due to the entry of Fc into
the GO surface (Fig. 4b). Also the presence of aggregated
nearly spherical Fe3O4 nanoparticles on the GO-Fc surface
is shown in Fig. 4c. FE-SEM images demonstrate that the
GO-Fc@Fe3O4 nanohybrid was successfully synthesized.
Furthermore, the EDX analysis of GO-Fc@Fe3O4 nanohy-
brid shows the elements in the structure of nanocatalyst.
EDX analysis is a powerful tool for evaluating the ele-
ments of the synthesized compounds. From EDX curves
(Fig. 5), they are confrmed that the related elements of
GO including C and O were present in the synthesized GO
(Fig. 5a). The EDX analysis of the over-oxidized form of
GO (GO-Epo) shows a signifcant increase of O element
(Fig. 5b). The presence of the abundance of epoxy rings on
the GO surface provides an option of using these active sites
for the covalently grafted of Fc moieties on the GO surface
through ring-opening reaction. The presence of Fe element
in the EDX analysis of GO-Fc (Fig. 5c) confrms the suc-
cessful modifcation of GO-Epo surface with Fc moieties.
Fig. 2 FT-IR spectra of GO, GO-Fc, GO-Fc@Fe3O4 nanohybrid
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
Fig. 4 FE-SEM images of a GO, b GO-Fc, c GO-Fc@Fe3O4 nanohybrid
Fig. 5 EDX curves of a GO, b GO-Epo, c GO-Fc, d GO-Fc@Fe3O4 nanohybrid
The EDX analysis of GO-Fc@Fe3O4 nanohybrid (Fig. 5d)
Also, the magnetic properties of GO-Fc@Fe3O4 nanocat-
confrms that the Fe3O4 nanoparticles are successfully grown
alyst have been studied by vibrating sample magnetometer
(VSM) technique at room temperature (Fig. 6). As shown
onto GO layers.
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
1463
basic sites including carboxylic acid, hydroxyl, Fc rings on
the GO-Fc surface (Fig. 7). Also, GO nanosheets provide a
large available surface area. These mentioned reasons can
promote the formation of desired products.
To determine the optimum reaction conditions, the
model reaction was performed under various conditions
including various solvents, temperature, and amount of
catalyst (Table 1). At frst, the model reaction was car-
ried out in various solvents and solvent-free systems in the
presence of 40 mg of synthesized nanocatalyst at ambient
temperature (Table 1, Entries 1–4). Among these condi-
tions, the best yield was achieved under solvent-free con-
ditions. Afterwards, the infuence of catalyst amount and
various temperature on the yield of the model reaction
was investigated by using diferent amounts of the synthe-
sized nanocatalyst (40, 30, 20, 10, and 5 mg) in the model
reaction. As indicated in Table 1, the best results were
obtained during 40 min under solvent-free conditions at
100 °C in the presence of 10 mg of the magnetic synthe-
sized nanocatalyst (Entry 13).
Fig. 6 Magnetic hysteresis loop of Fe3O4 and GO-Fc@Fe3O4 nano-
hybrid
in Fig. 6, the saturation magnetization (Ms) values of Fe3O4
and GO-Fc@Fe3O4 nanohybrid are 64 emu/g and 43 emu/g,
respectively. The saturation magnetization value is decreased
due to the intercalation of Fe3O4 nanoparticles into GO-Fc
layers. However, the GO-Fc@Fe3O4 nanocatalyst exhibited
enough magnetization value that can be easily separated
from the reaction mixture by an external magnet.
After fnding the optimum conditions, to explore the
catalytic activity, the reaction between 2-naphthol (1a) and
6-bromo-2-naphthol (1b) with various aromatic aldehydes
(2a–j), and malononitrile (3) (Scheme 2) were tested under
optimum conditions (Table 2). Moreover, the existence of
electron-donating and electron-withdrawing functional
groups on the aromatic aldehyde ring has no remarkable
diference in the yield of the reactions. These fndings dem-
onstrate the higher activity of the synthesized nanocatalyst.
Finally, we tested the recyclibility of GO-Fc@Fe3O4 as
a magnetic nanocatalyst for the coupling of three compo-
nents in the model reaction. Upon completion of the reac-
tion, the crude product was dissolved in hot ethanol and
the magnetic nanocatalyst was separated using an external
magnet. Then, the separated magnetic nanocatalyst was
washed several times with hot ethanol and dried. The
recovered magnetic nanocatalyst can be reused 7 times
without any signifcant changes in reaction yield (Fig. 8).
Catalytic test of GO‑Fc@Fe3O4 nanohybrid
To check the catalytic performance of the GO-Fc@Fe3O4
nanohybrid, frstly, a model reaction between 2-naphthol
(1a), benzaldehyde (2a), and malononitrile (3) was chosen.
According to previous studies, the presence of acidic and
basic sites in the synthesized nanocatalyst can facilitate a
coupling reaction between the above mentioned three com-
ponents [51, 76–78]. Also, based on the catalytic activity
of Fc, GO-Fc@Fe3O4 nanohybrid has several acidic and
Fig. 7 Schematic presentation
of the synthesized nanocatalyst
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
Table 1 Determining the optimum reaction conditions
Entry
Catalyst amount (mg)
Solvent
Temperature (°C)
Time (min)
Yield (%)
1
2
3
4
5
6
7
8
–
–
r.t
120
110
110
110
110
80
80
80
80
80
50
45
45
40
Trace
18
22
9
40
40
40
40
30
20
10
5
10
10
10
10
10
10
EtOH
MeOH
H2O
–
–
–
–
–
–
–
–
–
–
r. t.
r. t.
r. t.
r. t.
r. t.
r. t.
r. t.
r. t.
50
70
80
90
100
110
42
40
40
40
33
51
63
70
79
83
81
9
10
11
12
13
14
15
–
45
Reaction conditions: 2‐naphthol/benzaldehyde/malononitrile=1:1:1.2
Scheme 2 Synthesis of 2‐
amino‐3‐cyano‐4H‐pyran
derivatives (4a–l) under opti-
mum conditions
We propose a possible mechanism of GO-Fc@Fe3O4
catalyzed reaction as depicted in scheme 3. After activation
of the aldehyde groups, a Knoevenagel condensation was
carried out between aromatic aldehyde and malononitrile,
and arylidenemalononitrile was formed as intermediate.
Afterward, the arylidenemalononitrile was coupled with
2-naphthol through Michael’s addition reaction followed
by electrophilic cyclization to form the desired product [70].
hybrids analogs and hAChE inhibitors have been synthe-
sized by the AlCl3 promoted Friedländer-type reaction
between 2‐amino‐3‐cyano‐4H‐pyrans derivatives (4a–l) and
cyclohexanone as the selected ketone in dry 1,2-dichloroeth-
ane (DCE) at 83 °C, in good yields (50–92%) (Scheme 4)
[26, 66].
hAChE inhibition
Preparation of tacrine‑naphthopyran hybrid deriva‑
tives (5a–l)
One way to treat AD is inhibition of ChE enzymes. The
cholinesterase inhibition activity of compounds 4a–l and
compounds 5a–l was investigated in vitro and using Ell-
man’s spectrophotometric method against hAChE and hBu-
ChE [67]. Fresh blood was used to prepare the enzymes
14-Aryl-10,11,12,14-tetrahydro-9H-benzo[5,6]
chromeno[2,3-b]-quinolin-13-amines (5a–l) as new tacrine
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
1465
Table 2 Synthesis of 2‐
Entry
1
Product
Structure
Time (min)
40
Yield (%)
83
Obs. M.p. (°C)
278–281
Lit. M.p. (°C)
279–282 [51]
amino‐3‐cyano‐4H‐pyran
derivatives
4a
2
3
4b
4c
25
35
75
75
186–189
271–274
183–185 [51]
270–273 [76]
4
5
4d
4e
40
25
73
80
217–219
206–209
218–220 [76]
207–209 [76]
6
7
4f
30
25
69
78
269–272
210–213
271–272 [77]
212–215 [76]
4 g
8
9
4 h
4i
35
35
70
68
197–201
221–225
198–201 [78]
224–227 [78]
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
Table 2 (continued)
Entry
10
Product
Structure
Time (min)
20
Yield (%)
65
Obs. M.p. (°C)
256–260
Lit. M.p. (°C)
258–262 [78]
4j
11
12
4 k
4 l
25
40
70
50
228–232
256–259
228–229 [77]
258–260 [78]
[68]. The amount of inhibition of compounds 4a–l against
hAChE and hBuChE was not sufcient to obtain IC50 for
them and these compounds did not show inhibitory activity.
Tacrine and galantamine are used as a positive control and
the outcome was illustrated as IC50 amount of synthesized
compounds in Table 3. The obtained results have shown that
compounds 5a–l had medium inhibition activity of hAChE
at micro-molar. However, the amount of inhibition of these
compounds (5a–l) for hBuChE was not sufcient to obtain
IC50 for them and it can be said that these compounds do not
show hBuChE inhibitory activity. The range of IC50 values
was obtained from 0.16 µM to 37.48 µM for hAChE. Among
them, compound 5f with IC50 value equal 0.16 µM displayed
good inhibitory activity against hAChE which shows more
inhibitory activity of tacrine and galantamine.
Kinetic analysis
The mechanism of hAChE inhibition for compounds 5a–l
was investigated using compound 5f. So, kinetic studies
of compound 5f as a representative compound with the
greatest inhibit hAChE activity between other synthesized
compounds was carried out to characterize a better under-
standing of the mechanism of enzyme inhibition. Due to the
structural similarity of the synthesized compounds, a similar
practical method was predicted. Using the Lineweaver–Burk
plots was determined the type of inhibition (Fig. 9). Accord-
ing to the results, compound 5f was showed mixed-type inhi-
bition. This analysis gives inhibition constants with Ki value
equal to 0.8 µM and KI value equal to 0.3 µM. According to
available reports, compound 5f is presumably binding simul-
taneously to the CAS and PAS of hAChE [17, 22].
Molecular docking
The use of computational methods is interest to design new
compounds nowadays and the use of molecular docking stud-
ies can be obtained interaction afnity between molecules and
hAChE enzyme. So, molecular docking studies on the selective
compound 5f was performed, to get more information of inter-
act with hAChE on their active site. According to the results,
the derivatives can inhibit the enzyme, by binding to the
active site of hAChE. The results obtained from the docking
Fig. 8 Reusability of GO-Fc@Fe3O4 nanocatalyst during 7 runs
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
1467
Scheme 3 Sugested mechanism for the synthesis of 2‐amino‐3‐cyano‐4H‐pyran derivatives
Scheme 4 Preparation of
tacrine-naphthopyran hybrid
derivatives (5a–l)
of compound 5f (Fig. 10) show that this compound has a bet-
ter interaction than tacrine (Fig. 11). The binding energy get
for compound 5f and tacrine was equal to −11.61 kcal/mol
and −7.17 kcal/mol, respectively. The compound of 5f and
tacrine do not show any hydrogen bonding. The results showed
tacrine interacted with TYR341, PHE338, PHE295, TYR124,
SER293, TRP286, and VAL294. So, the results were obtained
for compound 5f show that TRP86, GLY448, SER203,
HIS447, and TYR337 placed around of naphthopyran moi-
ety, GLY121, ASP74 and SER125 are around of 2-Cl-ph and
PHE297, PHE295, TYR124,TYR341, and TRP286 placed
around of 5,6,7,8-tetrahydroquinolin-4-amine moiety. The
compound 5f interacted with the peripheral anionic site (PAS)
and catalytic active site (CAS) of hAChE.
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
Table 3 IC50 (μM) values for inhibitory activity of hAChE by the
synthesized tacrine-naphthopyran hybrid derivatives (5a–l)
Entry
Product
R
R´
hAChE (μM)
1
2
3
4
5
6
7
8
5a
H
H
H
H
H
H
H
Br
H
Br
H
H
–
H
4.25 0.32
37.48 1.33
10.95 1.03
12.23 0.67
2.03 0.4
0.16 0.07
2.66 0.32
1.69 0.11
0.23 0.2
25.60 1.03
29.31 1.23
8.02 0.71
0.19 0.05
5b
4-NO2
4-CH3
(CH3)2CH
4-Cl
5c
5d
5e
5f
2-Cl
5g
4-Br
3-Br
3-Br
4-F
4-F
2-Thiophene
–
5h
9
5i
10
11
12
13
14
5j
5k
5l
Tacrine
Galantamine
–
–
3 0.18
Fig. 10 Compound 5f interaction with hAChE
Conclusion
In summary, the chemically modifed GO-Epo by Fc moi-
ety was synthesized successfully. Then, the novel GO-Fc@
Fe3O4 nanohybrid was formed by synthesized of Fe3O4
nanoparticles onto the GO-Fc sheets. Various analyses
such as FT-IR, FESEM, EDX, XRD, and VSM analysis
were used for the evaluation of all of the intermediates to
fnal nanocatalyst. GO-Fc@Fe3O4 nanohybrid was used
as an efcient heterogeneous catalyst for the synthesis
2‐amino‐3‐cyano‐4H‐pyran derivatives (4a–l). Also, the
results of its catalytic activity investigation showed that
the presence of acidic and basic sites in the synthesized
nanocatalyst increased catalytic activity in this reaction.
The relative simplicity of separation, high yields, recy-
clability and reusability are the advantages of using
Fig. 11 Tacrine interaction with hAChE
GO-Fc@Fe3O4 as a catalyst in this multi-component reac-
tion. Also, 14-aryl-10,11,12,14-tetrahydro-9H-benzo[5,6]
chromeno[2,3-b]-quinolin-13-amines derivatives (5a–l)
Fig. 9 Lineweaver–Burk plots
of hAChE inhibition kinetic of
compound 5f
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Journal of the Iranian Chemical Society (2021) 18:1455–1470
1469
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were investigated. These compounds showed selectively
good inhibition of hAChE and among them, compound 5f
was a stronger inhibitor of hAChE with IC50 value equal
to 0.16 µM. But these series did not show any inhibition
for hBuChE.
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Acknowledgements The authors would like to acknowledge the fnan-
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Afliations
Sakineh Mozafarnia1,2 · Reza Teimuri‑Mofrad1 · Mohammad‑Reza Rashidi2
1
2
Department of Organic and Biochemistry, Faculty
of Chemistry, University of Tabriz, Tabriz 51664, Iran
Research Center for Pharmaceutical Nanotechnology, Tabriz
University of Medical Sciences, Tabriz, Iran
1 3